Boletín de la SEA - Sociedad Entomológica Argentina | SEA
Transcription
Boletín de la SEA - Sociedad Entomológica Argentina | SEA
N° 26 (1) 2015 Edición especial ISSN 1666-4612 Boletín de la SEA Sociedad Entomológica Argentina Encontranos en: http://seargentina.myspecies.info/ Facebook: Sociedad-Entomologica-Argentina http://www.museo.fcnym.unlp.edu.ar/ De los Editores Estimados lectores: Este número especial del Boletín, está íntegramente dedicado al VIII International Symposium on Phlebotomine Sandflies (ISOPS VIII). Esta publicación incluye 249 resúmenes de los trabajos presentados por los participantes del simposio, que fueron sometidos a arbitraje oportunamente. Los resúmenes están agrupados en dos bloques, uno correspondiente a las mesas redondas y ponencias orales, y otro a los posters. En todos los casos se abarcan diferentes temáticas, incluyendo taxonomía, filogenia, genética, ecología, control de vectores, entre otros. Las búsquedas se podrán realizar a través de un índice temático y/o por autor. Por último se muestra una tabla con información referida a la cantidad de autores por país y a la de presentaciones según cada modalidad. Esperamos que este número sea del interés de los lectores y agradecemos a los organizadores del Simposio la elección de nuestro Boletín para publicar los contenidos del mismo. Hasta el próximo número Las editoras Dirección y edición Nora Cabrera Fabiana Gallardo Edición y diseño Julia Rouaux Dirección Boletín de la SEA. División Entomología, Museo de La Plata. Paseo del Bosque s/n. La Plata CP(1900), Buenos Aires, Argentina. A los autores A todos aquellos que deseen enviar contribuciones para el Boletín por favor dirigirse previamente a Nora Cabrera: [email protected] para recibir las pautas editoriales. Dear Friends and Colleagues, It is a great privilege for us to host and organize the VIII International Symposium on Phlebotomine Sandflies in Puerto Iguazú, Argentina. This time the ISOPS features a Keynote Lecture by Dr. Eunice A. Bianchi Galati, 8 speakers at Round Tables and 73 Oral Presentations. In addition, 249 abstracts were presented, belonging to 813 authors from 35 countries. As many of you know, because you have participated in this process, the first ISOPS took place in Rome in 1991 due to the impulse of Dr. Robert Killick-Kendrick and Dr. Michele Maroli, honoring the 300th Anniversary of the first-known illustration of a Phlebotominae sand fly by Filippo Bonanni. After this former event, ISOPS has alternated between the Old and the New World in Venezuela, France, Brazil, Tunisia, Peru, and Turkey. In 2014, this original idea reaches the eight Symposium in Argentina. After seven successful ISOPS we recognize ourselves as a growing community, but without losing the feeling of a periodical gathering of friends. Now we must preserve the heritage but also face the new challenges, and look forward with the main objective to reduce the sand fly-borne diseases burden in a changing world. New generations and methodologies are being incorporated; new voices were added to the multilateral global complexity; new knowledge and so new questions come with each ISOPS. As an ISOPS group, we can constitute a consolidated voice of sandfly researchers that could give evidencebased opinions, asked for or given to health policy makers. The consensual outcomes should include the voice of leading experts but also the amazing experience of worldwide distributed groups. As the ISOPS event, we can periodically produce a document that updates research-based evidence, gaps and priorities, beyond objectives narrowed by individual, laboratory, country or 'technological fashion' interests. As the ISOPS community, we can strength the collaborative network by searching for multi-partner projects and cross-training empowerment internships, according to the priorities defined in the ISOPS document. We can also establish common ground to define basic research protocols in order to make the results of different groups more comparable, and so achieve faster conclusions and stronger recommendations. We thought of many ways to homage the respected and beloved ISOPS friends that we have lost in the last years: Dr. Bob Killick Kendric, Dr. Italo Sherlock, Dr. Richard Titus, and our friend Dr. Alexandre Peixoto. Finally, we realized that the better way to make so is to keep the ISOPS going on and growing. Therefore, we can further discuss the ideas presented here during ISOPS VIII. We are very thankful to the Ministry of Health of Argentina, to all conference sponsors for their contributions, to the members of the Honorary and Scientific Committees for their active participation, and to you for your willing attendance. We hope you have a fruitful ISOPS, and a pleasant stay in Puerto Iguazú, one of the seven Natural Wonders of the World. On Behalf of the Organizing Committee and the Argentinean Network for Research on Leishmaniasis (REDILA) Oscar Daniel Salomon Maria Gabriela Quintana Maria Soledad Santini Honorary President Eunice A B Galati HONORARY COMMITTEE Añez Néstor Killick Kendrick Mireille Marolli Michele Özbel Yusuf Pérez Enrique Rioux Jean-Antoine Ready Paul D Shaw Jeffrey SCIENTIFIC COMMITTEE Bates Paul Brazil Reginaldo Depaquit Jérôme Elnaiem Dia-Eldin Feliciciangeli Dora Ferro Cristina Galati Eunice Gallego Montserrat Lawyer Phillip G Léger Nicole Pimenta Paulo Rangel Elizabeth Valenzuela Jesús Volf Petr Warburg Alon Yaghoobi-Ershadi Reza ORGANIZING COMMITTEE President Oscar Daniel Salomón Collaborators Soraya Acardi Mariana Manteca Acosta Secretaries Analia Araujo Ma. Gabriela Quintana Alicia Paola Benitez Ibalo Ma. Soledad Santini Pablo Berrozpe Christina McCarthy Treasury Nicolás Silvero José Direni Mancini Magalí Giulinai Javier Liotta Accreditation and Diffusion Beatriz Oscherov Denise Fuenzalida Julia Portnoy Enrique Szelag Cristina Remondegui Mariela Martínez Lucrecia Villarquide Translations Sergio Casertano Graphic Design Claudia Nose María Eugenia Utgés Website Design Editorial and Publications Juan Rosa María Soledad Fernández Matías Parra Martín Abelardo ISOPS VIII Puerto Iguazú Argentina Content & Index N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina CONTENT ROUND TABLES AND ORAL SESSIONS Phlebotomine Sand Flies (Diptera: Phlebotominae) in Brazil” ID 1-O Albert Picado; Murari Lal Das; Mark Rowland; James Austin; Elisa De Lazzari. “Evaluation of Different Nets Against Phlebotomus Argentipes, The Vector of Visceral Leishmaniasis In Nepal” ID 2-O Rajesh B Garlapati, Sachidananda Samantaray, Mutum Ingobi Singh, Trey Barresi, Dylan Burruss and Richard Poche. “Multiple Fipronil Doses in Dairy Cattle for Vector Control in Bihar, India: Residue Study and Efficacy Determination Against the Sand Fly Phlebotomus Argentipes” ID 3-O M Derbali, L Polyakova, A Boujaâma, D Burruss, S Cherni, W Barhoumi, I Chelbi, R Poch, E Zhioua. “Laboratory and Field Evaluation of Rodent Bait Treated with Fipronil for Feed Through and Systemic Control of Phlebotomus papatasi” 6 ID 14-O V Carter, DP Bray, G Borges-Alves, E Dilger, O Courtenay, R Brazil and JGC Hamilton. “Sex Pheromones as a Pre-Mating Species Isolation Barrier in Brazil” ID 15-O Chukwunonso Onyemaechi Nzelu, Abraham G Cáceres, Ken Katakura, Yoshihisa Hashiguchi and Hirotomo Kato. “Identification of Peruvian Sand Fly Species (Diptera: Psychodidae) through DNA Barcodes” ID 16-O Randrianambinintsoa Fano José; Léger Nicole; Boyer Sébastien; Robert Vincent; Depaquit. Jérôme. “Origins and Affinities of Malagasy Phlebotomine Sandflies” ID 17-O Israel Cruz, Rosa Gálvez, Oihane Martín, Ivonne Pamela Llanes-Acevedo, Carolina Arcones, Rocío Checa, Ana Montoya, Carmen Chicharro, Silvia Miguelañez, Guadalupe Miró. “DNA Barcoding of Mediterranean Phlebotomine Sandflies Based on the Sequence Analysis of the Mitochondrial Cytochrome Oxidase I Gene” ID 4-O Mehmet Karakuş; Samiye Demir; Hüseyin Çetin; Suha Kenan Arserim; Önder Ser; Seray Töz; I Cüneyt Balcioğlu; M Kirami Ölgen; Bahtiyar Yilmaz; Yusuf Özbel.“Natural Infection and Insecticide Susceptibility Status of Wild Caught Sand Flies in Rural Areas of Antalya, Mediterranean Region of Turkey” ID 18-O Jérôme Depaquit, Maria Dolores Bargues, Santiago MasComa. “A Molecular Phylogeny of the Phlebotominae” ID 5-O David M Poché, William E Grant, Hsiao-Hsuan Wang. “Sand Fly Control in Bihar, India: Modeling Alternative Control Strategies Using Systemic Insecticides Orally Administered to Livestock” ID 19-O Aysheshm Kassahun, Kristýna Hlavačková, Vít Dvořák, Asrat Hailu, Alon Warburg, Jan Votýpka and Petr Volf. “Distribution, Phylogeny and Taxonomy of Sergentomyia in Ethiopia” ID 6-O Diwakar Singh Dinesh, Faizan Hassan, Vijay Kumar and Pradeep Das. “Development of Resistance Against DDT to Phlebotomus argentipes (Diptera: Psychodidae) and Search for Alternative Insecticide” ID 20-O Vit Dvořák, Jérôme Depaquit, Valentin Pflüger, Petr Halada, Kristýna Hlavačková, Veronique Lehrter, Jorian Prudhomme, Sonja Zapata, Petr Volf, Alexander Mathis. “Rapid Identification of Phlebotomine Sand Flies by Maldi-Tof Mass Spectrometry” ID 7-O Erika Santamaría, Olga Lucía Cabrera, Raul H Pardo. ”Toxic and Behavioural Effects in Experimental Huts of Long Lasting Insecticide Treated Nets Against Lutzomyia Longiflocosa in the Sub-Andean Rural Zone of Colombia” ID 21-O Vit Dvorak, Ozge Erisoz Kasap, Jerome Depaquit, Bulent Alten, Jan Votypka, Petr Volf. “Two New Species of the Subgenus Transphlebotomus Revealed by Molecular Taxonomy” ID 8-O Waleed Al Salem, Louise Kelly-Hope, Maha Abdeladhim, Salah Balgonaeem, Mohammed Al Zahrani, Jesús Valenzuela, Álvaro Acosta-Serrano. “A control strategy for Old World cutaneous leishmaniasis outbreaks” ID 22-O Ivonne Pamela Llanes-Acevedo, Carolina Arcones, Rosa Gálvez, Oihane Martín, Rocío Checa, Ana Montoya, Carmen Chicharro, Silvia Miguelañez, Susana Cruz, Guadalupe Miró, Israel Cruz. “DNA Sequence Analysis Suggests that Cytb-Nd1 Pcr-Rflp may not Be Applicable to Sandfly Species Identification in the Whole Mediterranean Region” ID 9-O Raúl H Pardo, Olga Cabrera, Catalina Marceló, Erika Santamaría. “Long Lasting Insecticide Treated Nets Decrease Sandfly Abundance and Blood Feeding Success Indoors in the SubAndean Region of Colombia: Results of a Hamlet Vector Control Trial after One Year Follow Up” ID 10-O Olga Lucía Cabrera, Erika Santamaría, Raúl H Pardo. “First Experimental Hut to Study The Effect of Insecticide Control Measures Against Sandflies (Diptera: Psychodidae)” ID 11-O Manteca Acosta M; Molina J; Utgés M E; Mastrangelo AV; Pérez AA; Santini M S; Salomón OD. “Efficacy Of Impregnated Bednets And Species Composition In Experimental Henhouses” ID 12-O Domenico Otranto; Filipe Dantas-Torres; Emanuele Brianti; Fabrizio Solari Basano; Dorothee Stanneck; Katrin Deuster.“Prevention of Canine Leishmaniosis in Naturally Exposed Dogs Using a 10% Imidacloprid/ 4.5% Flumethrin Collar (SERESTO®)” ID 13-O Andrey J de Andrade, Paloma H F Shimabukuro, Eunice A B Galati. “A New Proposal for the Taxonomic Identification of ID 23-O Reale S, Torina S, Zaffora G, Cosenza M, Pitti R, D'Agostino R, Intile S., Piazza M, and Vitale F. “Multilocus Real Time PCR Identification of Phlebotomine Sand Flies Captured in Southern Italy” ID 24-O Nicole Léger and Jérôme Depaquit. “The Genus Idiophlebotomus (Diptera; Psychodidae)” ID 25-O Añez Néstor, Rojas Agustina, Yépez José Yancarlos. “Sand Fly Species Transmitting Visceral Leishmaniasis at the Semiarid Region of Venezuela” ID 26-O Luis G. Estrada, Horacio Cadena, Edgar Ortega, Luz A Acosta, Andrés Vélez-Mira, Rafael J Vivero, Eduar E Bejarano, Sandra Uribe, Iván D Vélez. “Natural Breeding Sites of Lutzomyia Evansi (Diptera: Psychodidae) in the Urban Zone of Ovejas, Sucre – Colombia” ID 27-O Perez JE, D Rado, D Quispe, A Quispe, F Toccas, M Chacon, B Valencia, A Llanos, M Quispe, E Aguilar, R Pacheco & E Ogusuku. “Lutzomyia Faunas (Diptera: Psychodidae) in the East Side if the N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina Andean Cordillera in Cuzco, Peru” Özbel, P Volf, B Alten. “Phlebotomine Sand Flies Crossing Anatolia: Do Borders Limit their Distribution?” ID 28-O Karina Mondragon-Shem; Waleed Alsalem; Louise KellyHope; Maha Abdeladhim; Mohamed Alzahrani; Ziad Memish; Jesus Valenzuela; Alvaro Acosta-Serrano.”Differential Exposure to Sand Fly Bites in Saudi Arabia” ID 29-O Paulo Silva de Almeida, Alan Sciamarelli, Josué Raizer, Jaqueline Aparecida Menegatti, Sandra Cristina Negreli Moreira Hermes, Maria do Socorro Laurentino de Carvalho, Andrey José de Andrade, Rodrigo Gurgel-Gonçalves. “Predicting the Geographical Distribution of Leishmaniasis Vector Species in Central West Brazil” ID 30-O Raquel Gonçalves, Daniela Cristina Soares, Ricardo Guimarães, Walter Souza Santos, Gilberto Cesar de Sousa, Anadeiva Portela Chagas, Lourdes Maria Garcez. “Ecology of Sandflies in Risk Zones for Tegumentary Leishmaniasis Transmission in Santarém Municipality, Western Pará State, Brazil” ID 31-O Dia-Eldin A Elnaiem. “Phlebotomus Orientalis and the Fall and Rise in Visceral Leishmaniasis in East Africa” ID 32-O Bilel Chalghaf, Amine Toumi, Sadok Chlif, Jihene Bettaieb, Mariem Harrabi, Goze Bertin Bénié, Afif Ben Salah. “Distribution of Leishmaniasis Vectors in the Mediterranean Basin in the Light of Climate Change” ID 33-O Aviad Moncaz, Oscar Kirstein, Araya Gebresellassie, Wossenseged Lemma, Solomon Yared, Teshome Gebre-Michael, Asrat Hailu, Moshe Shenker and Alon Warburg. “Characterization of Breeding Sites of Phlebotomine Sand Flies in the Judean Desert and North-West Ethiopia”. ID 34-O Oscar David Kirstein, Araya Gebreselassie, Aviad Moncaz, Asrat Hailu, Teshome Gebre-Michael and Alon Warburg. “Fine Scale Analyses of Factors Contributing to the Transmission of Visceral Leishmaniasis in North Ethiopian Villages: An Eco-Epidemiological Approach” ID 35-O Wossenseged Lemma, Habte Tekie, Meshesha Balkew, Teshome Gebre-Michael3, Alon Warburg, Asrat Hailu. “SeroPrevalence L. donovani in Labour Migrants Population dnd Entomological Risk Factors in Extra-Domestic Habitats of MetemaHumera Lowlands - Kala-Azar Endemic Areas in the Northwest Ethiopia” ID 36-O Araya Gebresilassie1, Ibrahim Abbasi2, Oscar David Kirstein2, Aviad Moncaz2, Habte Tekie1, Meshesha Balkew3, Alon Warburg2, Asrat Hailu4, and Teshome Gebre-Michael3 “Host Preference Patterns of Phlebotomus (Larroussius) orientalis Parrot, 1936 (Diptera: Psychodidae) in the Visceral Leishmaniasis Endemic Area of Tahtay Adiyabo District, Northern Ethiopia” ID 37-O Esayas Aklilu, Habte Tekie, Meshesha Balkew, Alon Warburg, Asrat Hailu, Teshome Gebre-Michael. “Bionomics of Phlebotomine Sandflies in a Highland Kala-Azar Focus in LiboKemkem District, Northwestern Ethiopia” 7 ID 40-O M R Yaghoobi-Ershadi, M Darvishi, F Shahbazi, AA Akhavan, R Jafari, M Khajeian, H Soleimani, H Darabi, M H Arandian. “Epidemiological Study on Sand Flies in an Endemic Focus of Cutaneous Leishmaniasis, Bushehr City in Southwestern Iran” ID 41-O Jeffrey Jon Shaw. “Eco-Epidemiology of Leishmaniases: Vectors and New Scenarios of Transmission.” ID 42-O Alon Warburg. “Emerging Cutaneous Leishmaniasis in Israel: A Review of The Last Two Decades” ID 43-O Ns Singh; Doris Phillips-Singh; Ipe M Ipe; Shubham Pandey; Dinesh Lal. “Studies on the Sand Fly Fauna (Diptera: Psychodidae) in High-Transmission Areas of Visceral Leishmaniasis in North India, Special Emphasis on Uttaranchal Region” ID 44-O W Fares, RN Charrel, K Dachraoui, L Bichaud, W Barhoumi, M Derbali, S Cherni, I Chelbi, X de Lamballerie, E Zhioua. “Infection of Wild Sandflies Collected from Different BioGeographical Areas of Tunisia with Phleboviruses” ID 45-O Bongiorno G, Lisi O, Severini F, Vaccalluzzo V, Khoury C, Di Muccio T, Gradoni L, Maroli M, D'urso V, Gramiccia M, “Investigations On Sand Fly Bionomics and Leishmania Natural Infections in Eastern Sicily, Italy, with Particular Reference to Phlebotomus Sergenti” ID 46-O Alveiro Pérez-Doria, Margaret Paternina, Lily Martinez, Luis Paternina, Daniel Verbel-Vergara, Suljey Cochero, Eduar E. Bejarano, “Natural Infection of Lutzomyia Cayennensis Cayennensis (Diptera: Psychodidae) with Leishmania (Viannia) spp., Potential Vector in the Enzootic Cycle of Leishmaniasis in Urban And Rural Habitats of The Colombian Caribbean” ID 47-O Paul D Ready “Priorities for Progressing from Assessments of Vector Competence to the Transmission Modelling and Control of Leishmaniasis” ID 48-O Paulo F P Pimenta, Vanessa C Freitas, Nágila F C Secundino. “Leishmania Chagasi in its Natural Vector Lutzomyia longipalpis.” ID 49-O Jovana Sadlova, Veronika Hrobarikova, Jan Votypka, Alon Warburg and Petr Volf. “Xenodiagnosis of Leishmania donovani Infections in Asymptomatic Mice Using Phlebotomus orientalis Sand Flies ID 50-O V Seblova-Hrobarikova, J Sadlova, S Carpenter, P. Bates and P Volf. “Development of Leishmania Parasites in Biting Midges, Culicoides nubeculosus and Culicoides sonorensis (Diptera: Ceratopogonidae) and Implications for Screening Vector Competence” ID 38-O Solomon Yared, Meshesha Balkew, Alon Warburg, Asrat Hailu and Teshome Gebre-Michael. “Species Composition, Abundance and Seasonal Dynamics of Phlebotomus Species in a Visceral Leishmaniasis Endemic Area of Northwest Ethiopia” ID 51-O Luis Roberto Romero, Jorge Luis Rodríguez, Alveiro PérezDoria, José G. Vergara, Ana Montesino, Lily Paola Martínez, Luis Cortés, Suljey Cochero & Eduar Elías Bejarano. “The Sand Fly Lutzomyia evansi (Diptera: Psychodidae), as a Permissive Vector of Leishmania Infantum and Le. braziliensis In Bolívar, Colombia” ID 39-O O Kasap Erisoz, A Belen, C Alkan, F Gunay, V Dvorak, K Ergunay, S Aydın, J Votypka, A-L Banuls, R Charrel, A Özkul, Y ID 52-O Ana Clara Araújo Machado Pires, Paulo Filemon Paolucci Pimenta and Nágila Francinete Costa Secundino. “Experimental N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina Susceptibility of Lutzomyia (L.) Longipalpis to Different Species of Leishmania” ID 65-O Anayansi Valderrama Cumbrera, Mara Garcia Tavares, Ingrid Murgas, José Andrade Dilermando. “Genetic Assessment of Sandfly Populations in Panama” ID 53-O Shaden Kamhawi, Fabiano Oliveira and Jesus Valenzuela “The Prospect for a Sand Fly Saliva-Based Vaccine Against Leishmaniasis in Humans” ID 54-O Pitaluga AN, Tinoco B, Di Blasi T, Silva AM, Tempone A, Traub-Csekö YM. “Characterization of Lutzomyia Longipalpis Immune Responses with Emphasis in Leishmania Infection” ID 55-O Matthew E Rogers, Emilie Giraud, Rod J Dillon, Paul A Bates, Tamsyn Derrick, Oihane Martin, Ingrid Műller. “Promastigote Secretory Gel from Infected Sand Flies Subverts the Wound Healing Response in an Insulin Growth Factor1-Dependent Manner” ID 56-O Ranadhir Dey, Hamide Aslan, Amritanshu Joshi, Claudio Meneses, Pradeep K. Dagur, John Philip McCoy, Robert Duncan, Jesus G. Valenzuela, Hira L. Nakhasi and Shaden Kamhawi, “VectorTransmission of Leishmania donovani Triggers a Distinct Immune Response that Favors Parasite Survival and Dissemination” ID 57-O Wafa Rebai-Kammoun, Ikbel Naouar, Darragh Duffy, Valentina Libri, Milena Hasan, Amine Toumi, Afif B. Salah, Matthew Albert, Hechmi Louzir, Amel Meddeb-Garnaoui. “Cytokine Levels in Response to Salivary Gland Extracts and to Leishmania major Parasites in Individuals Living in Endemic Areas for Zoonotic Cutaneous Leishmaniasis in Tunisia” ID 58-O Ikbel Naouar, Amel Meddeb-Garnaoui, Amine Toumi, Nabil Belhadj Hamida, Afif B Salah, Hechmi Louzir. Presenting author at the event: Wafa Rebai Kammoun. “Prospective Study of a Cohort of Individuals at Risk of Zoonotic Cutaneous Leishmaniasis in Tunisia to Evaluate Clinical and Immunological Parameters and Identify Correlates of Protection.” ID 59-O Tatiana Di-Blasi; Antonio Jorge Tempone; Erich Loza Telleria; Fabricio K Marchini; José Marcelo Ramalho-Ortigão; Yara Maria Traub-Csekö. “The Possible Role of a Flagellar Protein from Leishmania sp. (FLAG) in the Parasite - Sand Fly Interaction” ID 60-O Votypka J, Pruzinova K, Hlavacova J, Volf P. “The Effect of Temperature and Avian Blood on Leishmania Development in Sand Flies” 8 ID 66-O Noteila M Khalid, Marium A Aboud, Fathi M Alrabba, DiaEldin A Elnaiem and Frederic Tripet. ”Evidence for Genetic Differentiation at the Microgeographic Scale in Phlebotomus papatasi Populations from Sudan” ID 67-O Hector M Diaz-Albiter, Caroline d Moraes, Maiara d Faria, Mauricio R Sant'Anna, Roderick J Dillon and Fernando A. Genta. “Gene Expression Profiling and Silencing of Key Digestive Enzymes in Lutzomyia Longipalpis” ID 68-O Remoli ME, Bongiorno G, Fortuna C, Marchi A, Bianchi R2, Khoury C, Ciufolini MG, Gramiccia M. ” Evaluation of Collection Methods for Phlebotomus-Borne Viruses Detection: Isolation and Viral RNA Integrity Performance” ID 69-O Andrea Martins da Silva, Antonio Jorge Tempone, André N Pitaluga, Fabricio K Marchini, Barbara Burleigh, Yara M TraubCseko. “Conditioned Media and Exosomes from Lutzomyia longipalpis LL5 Cells dsRNA-Transfected are able to Induce NonSpecific Antiviral Response in LL5 Naive Cells” ID 70-O Meneses C, Morales E, Rhodes C, Oliveira F, Valenzuela JG and Kamhawi S. “CD-Rom Sand Fly Fellas: The First Complete Video-Based Sand Fly Rearing Guide in High Definition” ID 71-O Vicente Estevam Machado; Thais Marchi Goulart; Arlene Gonçalves Corrêa; Mara Cristina Pinto. “Comparison of Volatile Compounds Emitted by Animals or Present in Plants on Attractiveness of Nyssomyia neivai (Diptera:Psychodidae) ID 72-O Ifhem Chlebi, Elyes Zhioua, and Gordon Hamilton. “Responses of the Sand Fly Phlebotomus papatasi Scopoli (Diptera: Psychodidae) to an Oviposition Pheromone Associated with Conspecific Eggs” ID 73-O Vivero J Rafael; Cadavid Gloria; Moreno H Claudia Ximena; Posada-López Laura; Sandra Uribe S. “Study on the Bacterial Midgut Microbiota Associated to Immature Stage of Genus Lutzomyia (Diptera: Psychodidae) Isolated from Natural Breeding Sites” ID 61-O Mary Ann McDowell and the Sand Fly Genome Consortium. “Two Sand Fly Genomes: Phlebotomus papatasi And Lutzomyia longipalpis” POSTERS ID 62-O Gloria I Giraldo-Calderón, Scott J. Emrich, Daniel Lawson, Frank H Collins and the members of the VectorBase consortium. “The Database Vectorbase and the Genomes of Lutzomyia longipalpis and Phlebotomus papatasi” ID 1-P Sirlei Franck Thies, Ana Lucia Maria Ribeiro, Jorge Senatore Rodrigues Vargas, Edelberto Santos Dias and Amilcar Sabino Damazo “Sandfly fauna (Diptera: Psychodidae) of urban area in the city of sinop, Mato grosso, Brazil” ID 63-O Samuel Tadros, Mariha Wadsworth, Mallory Hawksworth, RJ Nowling, Mary Ann McDowell, Gloria I Giraldo-Calderón. “Manual Annotation of G Protein–Coupled Receptors (GPCRs) in the Genomes of Lutzomyia longipalpis and Phlebotomus papatasi” ID 2-P Sirlei Franck Thies, Marinês Browers “Sandfly fauna (Diptera: Psychodidae) from regional of health in Sinop, Mato Grosso, Brasil, year 2013.” ID 64-O Gabriela Vela, Gabriel Trueba, Jérôme Depaquit, Sonia Zapata. “Genetic Variation of Nyssomyia trapidoi (Diptera: Psychodidae) in Three Endemic Zones of Leishmaniasis in the Ecuador” ID 3-P Lorena G. Caligiuri, Enrique A. Sandoval, María Soledad Santini, Soraya Acardi, Oscar D. Salomón, Lilian Tartaglino and Christina B. McCarthy “Testing the versatility of a Psychodiella sp. diagnostic assay in field surveys” ID 4-P Inés Martín-Martín, Ricardo Molina and Maribel Jiménez “Kinetics of anti-Phlebotomus perniciosus saliva antibodies in mice and rabbit será” N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 5-P Laura W. Juan, Ignacio T. Gould, Oscar D. Salomón, Juan Molina, Raul A. Alzogaray and Eduardo N. Zerba “Field evaluation of an insecticide formulated with active ingredients acting on larval and adult stages of Lutzomyia longipalpis” Visceral Leishmaniasis in northeast of Brazil: Fortaleza, state of Ceará, Brazil” ID 6-P Douglas De Almeida Rocha, Thaís Oliveira Coelho, Rodrigo Gurgel Gonçalves, Andrey José de Andrade and Marcos Takashi Obara “Characterization of susceptibility of populations of Lutzomyia longipalpis (Diptera: Psychodidae) to alphacipermetrin” ID 7-P Tobin Rowland, Silas Davidson, Kevin Kobylinski and Edgar Rowton “Effects of Leishmania major Infection on Sand Fly Feeding Responses to DEET and or permethrin treated uniform material” ID 8-P Horacio Cadena, Edgar Ortega, Luz A. Acosta, Andrés Vélez-Mira, Rafael J. Vivero,Eduar E. Bejarano, Sandra Uribe, Hernando Gómez, Luis G. Estrada and Iván D. Vélez “Soil analysis and vegetation associated with the breeding sites of Lutzomyia evansi in the municipality of Ovejas-Sucre a focus of leishmaniasis in Colombia” 9 ID 17-P Fernandes Brilhante A., Galati E. A. B, de Oliveira Cardoso C., Progênio dos Santos N., Ferreira de Souza J. and Vidal J. “Phlebotominae captured in area of transmission of American cutaneous leishmaniasis (ACL) in Xapuri municipality, Acre state, Western Brazilian Amazonian. Preliminary results” ID 18-P dos Santos Afonso M. M., de Miranda Chaves S. A., de Avelar Figueiredo Mafra Magalhães M., Garrido R., Graser C., de Carvalho B. M., Costa S. M. and Rangel E. F. “The geographic expansion of American Visceral Leishmaniasis associated with deforestation in Tocantins State, Brazil” ID 19-P dos Santos Afonso M. M., de Miranda Chaves S. A., de Pita Pereira D., da Costa Rego T. A., de Santana A. L. F., dos Santos Silva J., de Carvalho B. M., da Costa S. M., Giordano C. and Rangel E. F.“Lutzomyia (Lutzomyia) longipalpis feeding habits and natural infection by Leishmania (Leishmania) infantum chagasi in some Brazilian endemic areas of American visceral leishmaniasis” ID 9-P David Poché, Alex Gendernalik, Larisa Polyakova, Richard Poché, Madina Kozhahmetova, Batirbek Aimakhanov, Ziyat Abdeliyev and Bakhit Atshabar “The use of systemic insecticides to control Phlebotomus spp. sand flies in southern Kazakhstan” ID 20-P Stramandinoli Moreno E., Posteraro Freire M., Cordeiro Nascimento L. O., Zanini V., Ribeiro Alves V. , Fernandes Shimabukuro P. H. “Sand flies (Diptera, Psychodidae, Phlebotominae) collected in the Wajãpi Indigenous Reserve, Eastern Amazon, Brazil” ID 10-P José Carlos C B Ribeiro, Antônio Menezes Filho, Daniela M Parente, Teresinha C Farias, Danielle A Zacarias, Jalthon C da Silva, Dorcas L. Costa, Ivete L Mendonça, Carlos H N Cost “Transmission of Leishmania infantum to Lutzomyia longipalpis by humans with and without HIV infection, symptomatic and asymptomatic” ID 21-P Fernández M. S., Santini M. S., Cavia R. and Salomón O. D. “Lutzomyia longipalpis (Leishmania infantum vector) and Nyssomyia whitmani (Leishmania braziliensis vector) in the city of Puerto Iguazú, northeastern Argentina: seasonal abundance and relationship with domestic animals” ID 11-P Quintana MG., Santini MS, Britez N, Rojas de Arias A, Thomaz-Soccol V, Bisetto A, Bazzani R, Yadon Z, Salomón OD and the IDRC Project #107577 team “Addressing the emergence and spread of leishmaniasis in the borders of Argentina, Brazil and Paraguay” ID 22-P Machado C. A., Ribeiro Alves V. and Fernandes Shimabukuro P. H. “Geographical distribution of Nyssomyia Barretto, 1962 in Brazil” ID 23-P Rosa J. R., Parras M. A., Szelag E. A and Salomón O. D. “Vertical and horizontal distribution of sand flies in an area of Tegumentary Leishmaniasis in Chaco, Argentina” ID 12-P Fitipaldi Veloso Guimarães V. C., Lemos Costa P., da Silva F. J., Lopes de Melo F., Dantas-Torres F., Gomes Rodrigues E. E. and Pinto Brandão-Filho S. “Molecular detection of Leishmania in phlebotomine sand flies in a cutaneous and visceral leishmaniasis endemic area in northeastern Brazil” ID 24-P Parras M. A., Rosa J. R., Szelag E. A. and Salomón O. D. “Natural breeding sites of Phlebotominae (Diptera: Psychodidae) in a flood zone of Chaco, Argentina” ID 13-P Menezes, J.A., Sousa A. M., Morais M. H. G., Andrade-Filho J. D., Lima F. P., Fux B. and Margonari C. “Study of sand fly fauna in recent area transmission of leishmaniasis in the state of Minas Gerais, Brazil” ID 25-P Carvalho G.M.L., Quaresma P. F., Gontijo C. M. F., Rego F. D., Rugani J. M. N., Dias T. A. and Andrade Filho J. D. “Analysis of sand fly (Diptera: Psychodidae) bloodmeals using sequencing technique: test of sand fly experimentally fed” ID 14-P de Andrade A. J. and Gurgel-Gonçalves R. “New record and update on the geographical distribution of Pintomyia monticola (Costa Lima, 1932) (Diptera: Psychodidae: Phlebotominae) in South America” ID 15-P Rocha Dos Santos C. F., Lima M. C., Da Rocha Neto G. B. and Moreira Macena A. L. “Entomological monitoring of the programme of visceral leishmaniasis in Maceió, Alagoas State, Brazil: indicators and environmental risk factors” ID 26-P Acel A. M., Ribeiro A. L. M., Miyazaki R. D. and Senatore Vargas Rodrigues J. “Sandfly survey in ecotourism region in the district of Bom Jardim from the City of Nobres, Mato Grosso – Brazil” ID 27-P Ribeiro A. L. M., Senatore Vargas Rodrigues J., Parreiras de Almeida T., Marques de Souza E., Pereira Barreto Bettiol F. K., Acel A. M, Leal dos Santos F. A., Franck Thies S. and Dunjko Miyazaki R. “Leishmaniasis sandfly diversity in a rural area of the Cuiabá, Mato Grosso, Brazil” ID 16-P Silva R. A., Bevilaqua C. M. L., Rangel E. F., Santos F. K.M., Sousa L. C., Donato L. E. and Pessoa G. C. D. “Aspects of the ecology of Lutzomyia longipalpis and Lutzomyia migonei (Diptera: Psychodidae: Phlebotominae) in area of intense transmission of ID 28-P Dunjko Miyazaki R., Ribeiro A. L. M., Senatore Vargas Rodrigues J., Parreiras de Almeida T., Marques de Souza E., Pereira Barreto Bettiol F. K. and Acel A. M. “Incidence of sandflies at the zoological of Federal University of Mato Grosso, Cuiabá/MT-Brazil” N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 29-P Lemma W., Tekie H., Balkew M., Gebre-Michael, Warburg A. and Hailu A. “Population dynamics and habitat preferences of Phlebotomus orientalis in extra-domestic habitats of Metema Humera lowlands - kala-azar endemic areas in northwest Ethiopia” ID 42-P da Costa S. M., Cordeiro J. L. P, Afonso M.S.11 and Rangel E. F. “Habitat Suitability for Lutzomyia (Nyssomyia) whitmani and American Cutaneous Leishmaniasis (ACL) in Brazil” ID 30-P Lemos Costa P., Dantas-Torres F., José da Silva F., Fitipaldi Veloso Guimarães V. C., Gaudêncio K. and Pinto Brandão Filho S. “Ecology of Lutzomyia longipalpis in an area of visceral leishmaniasis transmission in north-eastern Brazil” ID 31-P de Oliveira Miranda D. E., Gaudêncio da Silva K., da Gloria Faustino M. A., Câmara Alves L., Pinto Brandão-Filho S., DantasTorres F. and de Carvalho G. A. “Diversity of phlebotomine sand flies in an american cutaneous leishamaniasis endemic area in northeastern brazil” ID 32-P Gaudêncio da Silva K., Silva de Morais R. C., de Oliveira Miranda D. E., de Arruda Moura A. C., Pinto Brandão-Filho S., de Paiva Cavalcanti M. and Dantas-Torres F. “A real-time PCR for characterization of multiple blood sources in field-collected phlebotomine sand flies” ID 33-P Maia C., Cristóvão J., Freitas F. B., Afonso M.O. and Campino L. “A three consecutive years phlebotomine sand fly survey in a zoonotic leishmaniasis endemic focus, the Algarve Region, Portugal” ID 34-PLemma W, Tekie H, Abassi I, Meshesha Balkew M., GebreMichael T., Warburg A. and Hailu A. “Nocturnal Activities and Host Preferences of P. orientalis in Extra-Domestic Habitats of Kala-Azar Endemic Metema–Humera Lowlands, North West Ethiopia” ID 35-P Rigg C., Valderrama A, Calzada J.E., Chavez L. F. and Saldaña A. “Prevalence of Leishmania spp. infection and host - blood feeding of sandflies transmitters of cutaneous leishmaniasis in an endemic locality in the district of capira, Panama” ID 36-P Arserim S.K., Karakus M. and Ozbel Y. “A study on nocturnal activity of sand flies in a leishmaniasis endemic village located in Aydin province of Turkey” ID 37-P Arserim S. K., Mermer A., Balcioglu I.C., Ermis V. O. and Ozbel Y. “Seasonal Activities and Ecological Features of Sand Flies (Diptera: Psychodidae: Phlebotominae) in Aydın Mountains and Surroundings Located in Western Turkey, Endemic Area for Cutaneous Leishmaniasis” ID 38-P Araujo A. V., Benítez Ibalo A. P., Rosa J. R., Oscherov E. B. and Salomón O. D. “Bionomy of Phlebotomine sandflies (Phlebotominae) in an outbreak area of visceral leishmaniasis in Corrientes, Argentina” 10 ID 43-P Gould, I. T., Fernández M. S., Pérez A A. and Salomón O. D. “A field study to evaluate the performance of two modified CDC light traps for collecting sandflies compared with the traditional CDC trap light” ID 44-P Gómez Bravo A., Gould I. T., Abril M., Salomón O. D. “Population composition and dynamics of Lutzomyia longipalpis and other Phlebotominae species in Clorinda, Northern Argentina” ID 45-P Acardi S. A, Steinhorst I. I., Santini M. S., Sandoval A. E., Salomon O. D. “Description of population dynamics and hourly activity of Lutzomyia longipalpis in the city of Posadas, Argentina” ID 46-P Mondragon-Shem K., Bell M., Suar M., Routray P., Posada L. and Cameron M “Comparison of CDC light and gravid traps for the collection of sandfly fauna in Bhubaneswar, Orissa (India)” ID 47-P Ilango Kandan “Taxonomy and diagnostic characters of the Phlebotomus argentipes species complex” ID 48-P Rodrigo E. Godoy, Eunice A. B. Galati, Nataly A. de Souza, Thiago V. dos Santos, Lindemberg Caranha de Sousa, Adriana Zwetsch and Elizabeth F. Rangel “Discriminant analysis as a possible tool to identification Sand flies (Diptera: Psychodidae) species through morphometric data” ID 49-P Jomaa Chemkhi, Souheila Guerbouj, Ikram Guizani and Afif Ben Salah “First report of abnormal numbers of spermathecae in a female Phlebotomus (Larroussius) longicuspis (Diptera: Psychodidae) in Tunisia” ID 50-P D. de Almeida Rocha, J. Barbosa C. Ferreira, R. GurgelGonçalves and Andrey J. de Andrade “First record of Psathyromyia pradobarrientosi (Le Pont, Matias, Martinez & Dujardin, 2004) (Diptera: Psychodidae: Phlebotominae) in Brazil” ID 51-P Priscila B. Sábio, Andrey J. de Andrade and Eunice A. B. Galati “Records of females Psathyromyia cuzquena Martins, Llanos & Silva, 1975 (Diptera, Psychodidae, Phlebotominae) in northern Brazil” ID 52-P Priscila B. Sábio, Andrey J. de Andrade and Eunice A. B. Galati “Assessment of the taxonomic status of Phlebotomus pifanoi Ortiz, 1972 (Diptera, Psychodidae, Phlebotomina ID 53-P Nil Rahola " Uncharted Phlebotomine sand flies in Gabon” ID 39-P Kirstein O. D., Faiman R., Gebreselassie A., Hailu A, GebreMichael T. and Warburg A. “Attraction of Ethiopian phlebotomine sand flies (diptera: psychodidae) to light and sugar-yeast mixtures (Co2)” ID 54-P Patrick F. Botelho, Fábio M. Medeiros da Silva, Rodrigo E. Godoy and Thiago V. dos Santos “Morphometric study on populations of Psychodopygus complexa and Ps. Wellcomei (Diptera: Psychodidae), vectors of cutaneous leishmaniasis in Brazil” ID 40-P Moreira de Carvalho B., Moncassim Vale M., Lorini M. L., dos Santos Afonso M. M. and Rangel E. F. “Ecological niche modeling predicts southward expansion of Lutzomyia flaviscutellata, vector of diffuse cutaneous leishmaniasis in South America, under climate change” ID 55-P Veracilda R. Alves, Claudio Ruy V. da Fonseca and Paloma H. F. Shimabukuro “Taxonomic review of Psychodopygus nangabeira, 1941 (Diptera, Psychodidae, Phlebotominae ID 41-P Kirstein o. D., Abassi I., Gebreselassie A., Gebre-Michael T., Hailu A. and Warburg A. “Identification of Plants fed upon by phlebotomine sand flies in Ethiopia” ID 56-P Priscila B. Sábio, Andrey J. de Andrade and Eunice A. B. Galati “Taxonomic revision of the subgenus Psathyromyia Barretto, 1962 (Diptera: Psychodidae: Phlebotominae)” ID 57-P Maia C., Parreira R., Cristóvão J., Afonso M.O. and Campino N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina L. “DNA Barcoding for the Identification of Sand Fly Species (Diptera, Psychodidae, Phlebotominae) in Portugal” longipalpis in henhouses in an American visceral leishmaniasis endemic area of São Paulo State” ID 58-P Casertano S.A., Andrade Filho J.D. and Salomon O.D. “New records of Phlebotominae (Diptera: Psychodidae) from the Atlantic Forest region of Argentina” ID 72-P Ferreira Souza C., Quaresma P. F., Dilermando Andrade Filho J., Peçanha Brazil R. and Dias Bevilacqua P. “Micropygomyia quinquefer (Dyar, 1929) (Diptera: Psychodidae: Phlebotominae): a puctative vector of Leishmania spp. ?” ID 59-P Pérez-Doria A., Romero-Ricardo L., Lastre N. and Bejarano Eduar E. “A method for extraction of genetic material which permits the integration of classical taxonomy with identification based on the DNA barcode” ID 60-P Cristian F. Souza, Reginaldo P. Brazil, José D. Andrade Filho, Paula D. Bevilacqua, Patrícia F. Quaresma and Mirelle B. Souza “Contribution to the knowledge of the fauna of sand flies (Diptera: Psychodidae: Phlebotominae) of Rio Doce State Park, Minas Gerais, Brazil” ID 61-P Amini Esfanjani MH, Barazesh A, Mazloumi Gavagni A and Saeidi Z “Sand fly fauna in Kaleybar districts, northwestern Iran” ID 62-P Mierez M. L., Rea M. J. F., Borda C. E. and Mosqueda L. A. “Malformations of the genitalia in male Micropygomyia quinquefer (Dyar, 1929) (Diptera: Psychodidae) in Corrientes, Argentina” ID 63-PPosada-López L., Vivero R., Uribe S. I., Acosta L. A., VélezMira A., Cadena H., Estrada L. G. , Ortega E., Bejarano E. and Vélez I. D. “The use of DNA Barcoding for taxonomic identification of the immature stages of phlebotomine sandflies in ovejas, sucre (Colombia)” ID 64-P Fernandez R., Zorrilla V., Espada L., Vásquez G. and Stell F. “Sand flies (Diptera: Psychodidae) from Peru: report of new records” 11 ID 73-P Kratochvílová T., Košťálová T., Kindlová M., Sumová P., Šíma M., Rohoušová I., Gramiccia M., Gradoni L., Hailu A., Baneth G., Warburg A. and Volf P.“Canine antibody response to Phlebotomus perniciosus and Phlebotomus orientalis bites” ID 74-P de Oliveira Lara-Silva F., Monteiro Michalsky E., Latorre Fortes-Dias C., de Oliveira Pires Fiuza V., Marques Pessanha J.E., da Silva S. R., Moreira de Avelar D., Alves Silva M., Vianna Mariano da Rocha Lima A.C., da Costa A. J. A., Machado-Coelho G. L. L. and Edelberto Santos Dias E. “Epidemiological aspects of vector, parasite, and domestic reservoir in area with no reported human cases of visceral leishmaniasis in Brazil” ID 75-P Monteiro Michalsky E., Valadão Lopes J., Oliveira Lara E Silva F., Alves SilvaM., Vianna Mariano Da Rocha Lima A. C., França-Silva J. C., Rodrigues Da Paixão F., Antunes Da Costa A. J. and Santos Dias E. “Sandflies fauna survey and DNA detection of Leishmania infantum in Lutzomyia longipalpis captured in the city of Sabara, Minas Gerais, Brazil” ID 76-Pde Nazaré Martins da Silva A., Ishikawa E. A. Y., Fadul M., Vasconcelos dos Santos T., Costa Vasconcelos dos Santos Y., Carvalho de Araújo P. T. and Queiroz Santos N. “Phlebotomine sand flies (Diptera: Psychodidae) in three municipalities of the state of Pará, Brazil, area of very high transmission of cutaneous leishmaniasis” ID 65-P Fuenzalida AD, Direni Mancini JM, Salomon OD, and Quintana MG “First record of the genus Pintomyia Costa Lima, 1932 (Diptera Phlebotominae) in the northwest of Argentina” ID 77-P Moreira Macena A L., Cavalcante De Matos Rodarte C., Tavares Da Silva M. E., Rocha Dos Santos C. F., Carneiro Lima M. and Braga Da Rocha Neto G. “Preliminary study of sandflies in the city of Maceió-Alagoas-Brazil” ID 66-P Reis, A. S., Saraiva, L., Gontijo, C.M.F., Andrade Filho, J. D. " Morphometric study of Lutzomyia longipalpis populations (Diptera: Psychodidae) collected in urban and environmental protected areas in Minas Gerais, Brazil” ID 78-P de Morais Oliveira A. F., Lima Batista H., Azevedo Leite K. C. and Nunes Lacerda H. M. “Education: demographic and social impacts on the rate of incidence of Visceral Leishmaniasis in the State of Tocantins, Brazil (2010)” ID 67-P Almazán M. C., Gil J. F., Chanampa M. M., Barroso P., Nasser J, Copa G. N., Oreste F., Castillo G. “Sand flies captures and identification of Leishmania subgenus in Giemsa-Stained slides of patients from five localities of Salta, Argentina” ID 79-P Silva Lana R., Monteiro Michalsky E., França Silva J. C., de Oliveira Lara e Silva F., Latorre Fortes Dias C., Martins J. and Dias E. S. “Eco-epidemiology of leishmaniasis in the region of Serra do Cipo, a major touristic centreof Minas Gerais” ID 68-P Szelag E. A., Rosa J. R., Parras M. A. and Salomón O. D “Phlebotominae Bionomics in the Humid Chaco Bio-region. Chaco, Argentina” ID 80-P de Moraes S. C., Lane Melo A. and Rangel E. F. “Survey of sandflies (Diptera: Psychodidade: Phlebotominae) in indoor environments, peridomestic and extra-domestic transmission area of American cutaneous leishmaniasis (ACL) and American visceral leishmaniasis (AVL) in Mato Grosso, Brazil” ID 69-P Szelag E. A., Parras M. A., Rosa J. R. and Salomón O. D “Dispersion of Lutzomyia longipalpis in the Chaco province, Argentina” ID 70-P Galvis Ovallos F., da Paixão Sevá A., Galucci E., Martins Soares R., Ferreira F. and Galati E. A. B. “Monthly distribution of Lutzomyia longipalpis in an endemic area of visceral leishmaniasis in São Paulo state, Brazil” ID 71-P Galvis Ovallos F., da Paixão Sevá A., Galucci E., Martins Soares R., Ferreira F. and Galati E. A. B. “Frequency of Lutzomyia ID 81-P de Moraes S. C., dos Santos L. H. M., Nunes Alexandre T. and Brito do Nascimento T. “Sandfly fauna (Diptera: Psychodidae) in endemic area for transmission of leishmaniasis and the relation of the presence of Lutzomyia (Lutzomyia) longipalpis and Lutzomyia (Nyssomyia) whitmani whit information demographics” ID 82-P Torina A., Reale S., Blanda M., La Russa F., Intile S., Lelli R., Vitale F. “Study of the abundance and the distribution of Sandfly in Sicily” N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 83-P Berrozpe P., Araujo A., Santini M. S., Utges M. E. and Salomon O.D. “Lutzomyia longipalpis distribution and abundance in the city of Corrientes, northeastern Argentina” ID 84-P de Oliveira Moura Infran J., Alovisi SouzaD., Fernandes de Souza W., Casaril A. E., Utida Eguchi G., Teruya Oshiro E., Eurico Fernandes C. and Gutierrez de Oliveira A.“ Study of Phlebotominae in modified white and black Shannon traps in a Maroon area at Piraputanga district, Aquidauna, Mato Grosso do Sul, Brazil” ID 95-P Dutra Rego F., Marteleto Nunes Rugani J., Quaresma P. F., Fernandes Shimabukuro P. H., Barbosa Tonelli G., Carvalho Scholte R. G., and Ferreira Gontijo C. M. “Molecular detection of Leishmania in sand flies (Diptera: Psychodidae) and spatial distribution of American Cutaneous Leishmaniasis in the Xakriabá Indigenous Reserve, Brazil” ID 85-Pde Oliveira Moura Infran J., Alovisi SouzaD., Fernandes de Souza w., Casaril A. E., Utida Eguchi G., Elia Teruya Oshiro E., Fernandes C. E. and Gutierrez de Oliveira A.“Nycthemeral rhythm of Phlebotominae in a Maroon area at Piraputanga district, Aquidauna, Mato Grosso do Sul, Brazil” ID 86-P Bustamante Gomez M., Diaz M., Espinoza J., Parrado R., Reithinger R. and Lineth Garciá A. “Sand Fly Fauna in Chapare, Bolivia: An Endemic Focus of Leishmania (Viannia) braziliensis” ID 87-PEspinoza Echeverria J., Bustamante M., Diaz M., Parrado R. and Lineth Garciá A. “Diversity and nycterohemeral activity of phlebotomines in San Julian community (Cochabamba, Bolivia)” ID 88-P Casaril A. E., Falcão de Oliveira E., Zandonaide Monaco N., Utida Eguchi G., Rezende de Figueiredo H.,Lopes Fontoura Mateus N., de Souza Fernandes W., Petilim Gomes S., Alovisi Souza D., Oliveira Moura Infran J., Teruya Oshiro E., Galati E. A. B. and Gutierrez de Oliveira A. “The Sandfly Fauna (Diptera: Psychodidae: Phlebotominae) in Corumbá, Mato Grosso do Sul, Brazil” ID 89-Pdos Santos Silva J., Pereira dos Santos A., Kássio Moura Santos F., Rodrigues da Silva L. O., Caranha L. and Rangel E. F. “Entomologic surveillance of Phlebotomine sand flies vectors of Leishmaniasis in areas impacted by the Transfer of the São Francisco River project, in the state of Ceará, Brazil” ID 90-P Gebresilassie A., Kirstein O. D., Moncaz A., Tekie H., Balkew M., Warburg A., Hailu A., and Gebre-Michael T. “The Influence of moon light and lunar periodicity on the density of Phlebotomus (Larroussius) orientalis Parrot, 1936 (Diptera: Psychodidae) from light trap collections in Ethiopia” ID 91-PRodríguez-Jiménez J., Pérez-Doria A., Romero-Ricardo L., Vergara J., Montesino A., Martínez L., Cortés L., Cochero S., and Bejarano E. E. “Characteristics of human dwellings and their surroundings that favour the abundance of phlebotomines in four urban areas of the Caribbean coast of Colombia” ID 92-P Barata I. da R., Pinheiro M. S. B., da Silva F. M. M., Feitosa Brandão R. C., dos Santos T. V. and Silveira F. T. “Phlebotomine sand fly fauna (Diptera: Psychodidae) and natural flagellate infections from An area highly endemic for cutaneous leishmaniasis on the Brazilian-Bolivian frontier in the state of Acre, Brazil” ID 93-P Franck Thies S., Ribeiro A. L. M., Senatore Rodrigues Vargas J., Djunko Miyazaki R., Monteiro Michalsky E. and Santos Dias E. “Sandflies (Diptera: Psychodidae) naturally infected by Leishmania in rural areas of the city of Nova Mutum, Mato Grosso, Brasil” ID 94-P da Silva T. R. R., Gobira Guimarães de Assis M. D., Dutra Rêgo F., Posteraro Freire M. and Fernandes Shimabukuro P. H. “Sand fly fauna (Diptera, Psychodidae) collected in the Middle Purus river region, Amazonas, Brazil” 12 ID 96-P Jomaa Chemkhi, Rihab Yazidi, Mahdi Driss, Jihene Bettaieb and Afif Ben Salah “First report of natural promastigote infection of Sergentomyia dreyfussi (Diptera: Psychodidae) sand flies in an endemic focus of cutaneous leishmaniasis in central Tunisia” ID 97-P Laurenti M. D., Francesquini F. C., Passero L. F., Tomokane T. Y., Carvalho A. K., Corbett C. E. P. and Silveira F. T. “Salivary gland homogenates from wild-caught Lutzomyia flaviscutellata and L. (Psychodopygus) complexus did not exacerbate Leishmania (L.) amazonensis and L. (V.) braziliensis infection in BALB/c mice” ID 98-P Antonio Marque Pereira Júnior, Carolina Bioni Garcia Teles, Ana Paula de Azevedo dos Santos, Felipe Arley Costa Pessoa, Jansen Fernandes de Medeiros. "Natural infection of phlebotomine sand flies (Diptera: Psychodidae) by Leishmania Ross (Kinetoplastida: Trypanosomatidae) in “terra firme” and “várzea” enviroments in Tefé municipality, Amazonas state, Brazil” ID 99-P Martín-Martín I, Jiménez M., González E. and Molina R. “Transmission of Leishmania infantum to hamsters through the bite of experimentally infected Phlebotomus perniciosus and monitoring of animals infectivity by xenodiagnosis” ID 100-P Bates P.A., Bates M.D. and Kwakye-Nuako G. “Phylogenetic analysis of the Leishmania enriettii complex and speculations on their vector biology” ID 101-P Fernandes M.F., dos Santos K. M., Ferrari Júnior A. C., Silva Levay A. P., Negrão F. J., Fernandes W. D., Oshiro E. T., Cavalheiros Dorval M. E. M, Andrade Filho J. D., Gutierrez de Oliveira A. and Galati Eunice A. B. “Vector competence of Nyssomyia whitmani (Diptera: Psychodidae: Phlebotominae) to Leishmania (Leishmania) amazonensis” ID 102-P Rado D., Pita-Pereira D., Araujo Pereira T., Quispe-Florez MM, Brahim L., Rangel E., Valladares B. and Pacheco R. “Natural Infection of Lutzomyia (Nyssomyia) y. yuilli and L. (Trichophoromyia) octavioi with Leishmania spp. in Aguas Calientes, La Convencion, Cusco, Perú” ID 103-P Quispe A., Quispe-Ricalde M. A., Rado D., Quispe-Florez MM, Soto M., Pérez J.E., Valladares B. and Pacheco R. “Lutzomyia (Nyssomyia) y. yuilli and L. (Trichophoromyia) sp. from Alto Ivochote, La Convención, Cusco, Peru” ID 104-P Cutolo A. A, Ovallos F.G., Oliveira C., Lavitschka, Dantas da Silva M., Molla L. M., dos Santos Nogueira F., Pereira-Chioccola V. L., Menz I. and Galati E. A. B. “Aggregated pattern of Leishmania infantum chagasi transmission from naturally infected dogs to laboratory reared Lutzomyia longipalpis through xenodiagnosis” ID 105-P Pech-May Angélica, Sánchez-García Laura, BerzunzaCruz Miriam, Becker-Fauser Ingeborg, Escobedo-Ortegón Javier, Rebollar-Téllez Eduardo A. " Vectors of Leishmania mexicana in southeaster of Mexico” ID 106-P Boussaa Samia, Ouanaimi Fouad, Kahime Koloud, N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina Echchakery Mohamed, Boumezzough Ali and Pesson Bernard “Genetic characterization of northern and southern populations of Phlebotomus sergenti (Diptera: Psychodidae) populations in Morocco” Pinheiro M. P., da Silva Lopes T. E., de Medeiros Silva M. M. and da Silva Junior J. B. “Popular knowledge of leishmaniasis transmission in a peri-urban area of the municipality of São Gonçalo do Amarante, Brazil” ID 107-P Noteila M Khalid, Marium A Aboud, Fathi M Alrabba, DiaEldin A Elnaiem and Frederic Tripet. "Evidence for genetic differentiation at the microgeographic scale in Phlebotomus papatasi populations from Sudan” ID 119-P Alves M. de L., Freire de Melo Ximenes M. de F, Fernandes de Araújo M. F., Gomes Pinheiro M. P., da Silva Lopes T. E., da Silva Junior J. B. and Gurgel Mendes I.“ Educational comic strip booklet as an instrument of scientific dissemination of leishmaniasis in the state of Rio Grande do norte, Brazil” ID 108-P Oliveira D. M. S., Lima J. A. N., Silva B. J. M., Farias L. H. S., Santos T.V., Silveira F.T and Silva, E. O. “Analysis of carbohydrate epitopes on the surface of midgut epithelium of Lutzomyia (Lutzomyia) longipalpis and Lutzomyia (Nyssomyia) antunesi (Diptera, Psychodidae) from Pará, State Amazonian Brazil” ID 109-P Bussoni Bertollo D. M., de Cassia Hilário Castilho R., Costa Nunes Soares M. M., Kindler Figueiredo J., da Silva Meneghello B. H.; Mitsuyoshi Hiramoto R. and Tolezano J. E. “Comparative analysis between canine visceral leishmaniasis and sandfly presence in the endemic municipalities” ID 110-P Nunes Peres Dias Q., Barbosa de Souza M., Corrêa Novo S. P., Antônio de Medeiros Meira and do Santo Ponte C. “Survey of sandfly vectors in areas surrounding Pedra Branca Massif, Municipality of Rio de Janeiro, Brazil” ID 111-P Ferrolho J., Gomes J., Alves-Pires C, Cristóvão J.M., Maia C., Campino L. and Afonso M.O. “Rotation of the external genitalia in male Phlebotomine sandflies (Diptera, Psychodidae) in laboratory conditions and in captured specimens in Algarve, Portugal” ID 112-P Vasconcelos dos Santos T., Batista Campos M., Aprígio Nunes Lima J., Feitosa Brandão R. C., Medeiros da Silva F. M., Barros Pinheiro M. S., da Silva Santos L. A., De Freitas Leão E., da Rocha Barata I. and Fernando Tobias Silveira “Some field and laboratory observations on the biology of Nyssomyia antunesi (Diptera: Psychodiadae), a suspected vector of Leishmania (Viannia) lindenbergi in Amazonian Brazil” ID 113-P Goulart T. M., Machado V. E. and Pinto M. C. “Attractiveness of different diets for Nyssomyia neivai (Diptera: Psychodidae) larvae” ID 114-P Goulart T. M., Machado V. E. and Pinto M. C. “Effect of Nyssomyia neivai (Diptera: Psychodidae) density on oviposition” ID 115-P Rita Mukhopadhyay, Goutam Mandal, Jose Orta, Srotoswoti Mandal, Claudio Meneses, Hamide Aslan Suau, Eric Beitz, Jesus Valenzuela, Marcelo Ramalho-Ortigao, Shaden Kamhawi " Role of aquaporins in the physiology of Leishmania vector Lutzomyia longipalpis” ID 116-P Dutra Rêgo F., de Souza Pinto I., Machado C. A., Melim Ferreira G. E., Cupollilo E. and Fernandes Shimabukuro P. H. “Leishmaniasis Virtual Laboratory (LVL) - a new tool for the study of American sand flies (Dipetra, Psychodidade, Phlebotominae)” ID 117-P Mohammad S. Al-Khalifa; Ibraheem M. Al-Hazza and M. Y. Lubbad “Immunological studies on the cutaneous Leishmaniasis in Al-Kharj Region, Saudi Arabia” ID 118-P Alves M. de L., Freire de Melo Ximenes M. de F., Gomes 13 ID 120-P Gomes Pinheiro M. P., Freire de Melo Ximenes M. de F., de Melo Pereira Pinheiro M. P., de Lima Alves M. “Cordel folk literature as a tool for preventing leishmaniasis in Brazil” ID 121-P Rea M. J. F., C Borda. E. “Survey of American tegumentary leishmaniasis in the northeast of Argentina during 20112014” ID 122-P Lozano A., Basmadjián Y., Vitale E., Satragno D., Canneva B., Verger L., Tort C., Viera A., Romero S., Ríos C., Lagarmilla P. "Canine Visceral Leishmaniasis. Imported cases in Uruguay, 20102014" ID 123-P de Freitas Rocha A. T., Melo Silva S., Araújo Soares M. R., Costa Fortier D., Silva Barbosa D. B., Nery Costa C. H. " Occurrence of Lutzomyia longipalpis and spatial distribution of visceral leishmaniasis in Floriano, Piauí, Brazil: preliminary data” ID 124-P Santos M. F. C., Paranhos Filho A. C., Gamarra R. M., Fernandes W. D.; Brazil R. P. and Oliveira A. G. “Use of Remote Sensing in the analysis of populations of Lutzomyia longipalpis complex” ID 125-P Utgés M. E., Fuenzalida D., Parras M., Gould I. T., Casertano, S. and Salomón O. D. “Geometric morphometry of northeastern populations of Nyssomyia neivai from Argentina” ID 126-P Rado D., Pérez J. E., Vilela M., Quispe D., Quispe A., Aguilar E. G., Rangel E., Valladares B., Pacheco R. “Sandflies (Diptera: Psychodidae) of Aguas Calientes, La Convención, Cusco, Perú” ID 127-P Toccas F., Chacon M., Pérez J. E., Rado D., Soto M., Mendoza J., Luna R., Aguilar E. G., Valladares B. and Pacheco R. “Lutzomyia spp. (Diptera: Psychodidae) in intra, peri and extradomiciliar environments in Yomentoni (right margin), La Convención, Cusco, Perú” ID 128-P Rezende de Figueiredo H., Casaril A. E., Infran J. O. M., Teruya Oshiro E., Rodrigues Mendes A., Andrade Filho J. D., Moraes Ribeiro L., Falcão Oliveira E., da Cunha Santos M. F. and Gutierrez de Oliveira A. “Ecology of Lutzomyia longipalpis (Diptera: Psychodidae) in Aquidauana, endemic area for leishmaniases, Mato Grosso do Sul, Brazil. Preliminary results” ID 129-P Antonio Marques Pereira Júnior, Eric Fabricio Marialva, Moreno de Souza Rodrigues, Felipe Arley Costa Pessoa, Jansen Fernandes de Medeiros. " Studies on fauna of phlebotomine sand flies (Diptera: Psychodidae) at “Terra firme” and “Várzea” environments in Tefé municipality, Amazonas state, Brazil” ID 130-P Jiménez M., González E., Hernández S. and Molina R. “Phlebotomine sand fly survey in the focus of human leishmaniasis in south western Madrid region, Spain: 2012-2013” N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 131-P Jiménez M., González E., Hernández S. and Molina R. “Molecular detection of Leishmania infantum and host-feeding preferences in Phlebotomus perniciosus from the focus of leishmaniasis in south western Madrid region, Spain: 20122013” P. C., de Almeida M. R. “Sandflies (Diptera, Psychodidae, Phlebotominae) in Pitoco cave, an archeological site in northern Mato Grosso do Sul state, Brazil” ID 132-P Zoghlami Z., Chouihi E., Barhoumi W., Dachraoui K., Massoudi N., Ben Helel K., Habboul Z., Hadhri§ M. H., Limam S., Mhadhbi M., Gharbi M., and Zhioua E. “Interaction between canine and human visceral leishmaniases in a holoendemic focus of Central Tunisia” ID 133-P Nilsa Gonzalez-Britez, Nilda Portillo, Maria Ferreira, Martha Torales, Nidia Martínez, Luciano Franco. "Leishmaniasis vectors in the Alto Paraná Department, Paraguay” ID 134-P da Silva D. F., de Castilho Sanguinette C., Barbosa Tonelli G., Tanure A., Vidal Stumpp R. G. A, Dutra Rêgo F., Gontijo C. M. F. and do Andrade Filho J. D. “Sand flies (Diptera: Psychodidae: Phlebotominae) collected in urban and wild area in the district of Barra do Guaicui, Várzea da Palma municipality, Minas Gerais, Brazil” 14 ID 144-P Santos Carvalho S. M., Fitipaldi Veloso Guimarães V. C., dos Santos P. R. B. “First report of Lutzomyia longipalpis in Uruçuca, cocoa zone of the southern state Bahia, Brazil” ID 145-P Ballart C., González E., Ravel C., Jiménez M., Abras A., Molina R., Portús M., Gállego M. “Identification of blood meals in Leishmania infantum vectors from the Northeast of Spain: Phlebotomus ariasi and P. perniciosus by Polymerase Chain Reaction based and sequencing methods” ID 146-P de Lima Alves M., de Melo Ximenes M. F. F., Pinheiro M. P. G., da Silva J. H. T., da Silva Lopes T. E., de Medeiros Silva M. M. “Phlebotomine sanflies (Diptera: Psychodidae) in a peri-urban setting in the metropolitan region of Natal, Brazil” ID 147-P Toccas F., Rado D., Quispe A., Quispe C., Galindo B., Chacon M., Quispe-Ricalde MA, Valladares B., Pacheco R. “The Sand Fly Fauna (Diptera: Psychodidae) in Alto Ivochote, La Conveción, Cusco, Perú” ID 135-P Neves Vianna E., Franco Morais M. H., Sobral de Almeida A., Chagastelles Sabroza P., Carneiro M. and Santos Dias E. “Evaluation of the abundance of phlebotomine sand flies in households from an area of Visceral Leishmaniasis transmission in Belo Horizonte, Minas Gerais, Brazil” ID 148-P Martínez M. F., Santini M. S., Utgés M. E., Kowalewski M., Salomón O. D. “Study of phlebotomines in two wild areas of northeastern Argentina and the potential association with howler monkeys (Alouatta caraya)” ID 136-P Cavalcante Lamy Serra e Meira P., Lacerda Abreu B., Lusardo de Almeida Zenóbio A. P., de Castilho Sanguinette C., Mayr de Lima Carvalho G., Saraiva L. and Andrade Filho J. D. “Phlebotominae sand fly fauna in two urban parks in Belo Horizonte, Minas Gerais, Brazil (Diptera: Psychodidae)” ID 149-P de Melo Ximenes M. F. F., Antonaci Gama R., da Silva Lopes T. E., Melo Freire R. C., de Lima Alves M., Pinheiro M. P. G., da Silva J. H. T., de Medeiros Silva M. M. “Phlebotominae sanflies (Diptera: Psychodidae) in a preserved area of the Caatinga biome in Rio Grande do Norte State, Brazil” ID 137-P Cavalcante Lamy Serra e Meira P. and Andrade Filho J. D. “Species of phlebotomine sand flies (Diptera: Psychodidae: Phlebotominae) reported in Brazilian caves” ID 150-P Basmadjián Y., Canneva B., Verger L., Vitale E., Lozano A., Sequeira C., Lozano A., Satragno D., Tort C., Viera A., Ríos C., Lagarmilla P. and Romero S. “Lutzomyia longipalpis in Bella Unión, Department of Artigas, Uruguay. One year of surveillance: July 2013- June 2014” ID 151-P Oliveira A.F.J., Freitas R.A., Pessoa F.A.C., Aguiar N.O. “Phlebotomine Sand Fly survey in Central Amazon, Amazonas State, Brazil with a new register of Lutzomyia caligata” ID 138-P Barbosa Tonelli G., Dutra Rego F., Mayr de Lima Carvalho G., Tanure A. and Andrade Filho J. D. “Ecology of sand flies (Diptera: Psychodidae: Phlebotominae) in the Natural Heritage Private Reserve of Serra do Caraça, Minas Gerais, Brazil” ID 139-P Tanure A., Barbosa Tonelli G., Dutra Rêgo F., Mayr de Lima Carvalho G., Gontijo C. M. F., Fontes Paz G. and Andrade Filho J. D. “Study of sand flies (Diptera: Psychodidae: Phlebotominae) in a transmission area of leishmaniasis in Minas Gerais, Brazil” ID 140-P Souza G. D., A. Cardoso, de Carvalho R. M. J. S. “Assessment of sand flies (Diptera: Psychodidae) in an area with canine visceral leishmaniasis in the municipality of Porto Alegre, Rio Grande do Sul, Brazil” ID 141-P Basmadjián Y., Canneva B., Verger L., Lozano A., Satragno D., Supparo E., Sequeira C., Romero S., Viera A., Tort C., Ríos C., Lagarmilla P. and Vitale E. “Persistence of Lutzomyia longipalpis, vector of Visceral Leishmaniasis, in the city of Salto, Uruguay” ID 142-P Verger L, Lozano A, Vitale E, Satragno D, Sequeira C, Canneva B, Basmadjián Y, Rios C, Lagarmilla P, Tort C, Viera A, Romero S. “Serological surveillance in dogs of the city of Bella Unión, a risk area for Visceral Leishamaniasis” ID 143-P Galati E. A. B., de Almeida P. S., de Paula M. B., Costa Filho ID 152-P de Arruda Moura A. C., da Silva K. G., D. de Oliveira Miranda E., Ramalho M., da Silva F. J., Brandão-Filho S. P., Otranto D., Dantas-Torres F. “Phlebotominae sanflies in military training area in north-eastern Brazil” ID 153-P Ferro Cr., Lopez M., Fuya P., Lugo L., Cordovez J. M., Gonzalez C. “Relating vector species potential distribution and richness to transmission foci of Cutaneous Leishmaniasis in Colombia” ID 154-P Ballart C., Pesson B., Martín-Sánchez J., Alves-Pires C., Morillas F., Afonso O., Portús M., Gállego M. “Risk of leishmaniosis transmission in the Pyrenean region of Spain assessed through the isoenzymatic analysis of potential sand fly vectors (Phlebotomus ariasi and P. perniciosus)” ID 155-P Barhoumi W., Qualls W. A., Archer R., Fuller D. O., Chelbi I., Cherni S., Derbali M., Arheart K. L., Zhioua E.and Beier J. C. “Irrigation in the arid regions of Tunisia impacts the abundance and apparent density of sand fly vectors of Leishmania infantum” N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 156-P Pinheiro M. P. G., de Melo Ximenes M. F. F., de Lima Alves M., da Silva Júnior J. B., da Silva J. H. T., M de Medeiros Silva. M., da Silva Lopes T. E. “ Sandflies (Diptera: Psychodidae: Phlebotominae) from the Nisia Floresta National forest, metropolitan region Natal, Rio Grande do Norte State, Brazil” ID 170-P Erin Dilger, Orin Courtenay and Graham Medley “Density dependence in host-vector relationships and consequences for transmission” ID 157-P Erisoz Kasap O., Alten B. “Entomological Survey of Phlebotomine Sand Flies (Diptera: Psychodidae) in Turkey” ID 158-P Copa G. N., Marco J. D., Nasser J. R., Gil J. F. “Distribution of sandflies species in a locality endemic for American tegumentary leishmaniasis from the north of Argentina, bordering with Bolivia” ID 159-P Ruiz-Lopez F., Cadena H., Vélez Mira A., Posada-López L., Contreras A., Tirado Duarte D., Vélez I. D. “Effectiveness of a rapid multidisciplinary protocol for the epidemiological evaluation of an active focus of cutaneous leishmaniasis: Remedios, Antioquia, Colombia” ID 160-P Villarquide M. L., Fernández M. S., Santini M. S., Salomón O. D. “Abundance Association of Nyssomyia whitmani with the number of farm animals and its dwelling distance to the house, in areas with Tegumentary Leishmaniasis transmission in Puerto Iguazú, Misiones, Argentina” ID 161-P Ferro C., Fuya P., Lugo L., González C. “Biodiversity of Phlebotominae subfamily (Diptera: Psychodidae) in the Colombian Amazon and Orinoco” ID 162-P Enriquez S., Ariviliaga J., Vaca F., Benitez Ortiz W., Kirstein O., Abassi I., M Calvopiña., Pozo K., Romero V., O Chiluisa., E Villacrés and Warburg A. “Sand fly vectors of cutaneous leishmaniasis on the Western slopes of the Ecuadorian Andes” ID 163-P Direni Mancini J. M., Fuenzalida A. D., Saracho Bottero M. N., Lizarralde de Grosso M. S., Salomón O. D., Quintana M. G. “Abundance of Phlebotominae in endemic area of cutaneous leishmaniasis, Tucumán, Argentina” ID 164-P Ouanaimi F., Boussaa S., Kahime K., Echchakery M., Boumezzough A. “Phlebotomine sandflies (Diptera: Psychodidae) of Morocco: Results of entomological surveys along the OuarzazatM'hamid, Zagora-Chefchaouen-Nador and Zagora-Tata-Marrakesh roads” ID 165-P Kahime K., Boussaa S., Ouanaimi F. and Boumezzough A. “Spatial relationship between environmental factors and sand fly (Diptera: Psychodidae) distribution associated with leishmaniasis risk in centre & southern Morocco” ID 166-P Boussaa S., Ouanaimi F., Boumezzough A. “Identification and characterization of larval lodging of phlebotomine sand flies (Diptera: Psychodidae) in Marrakech, Morocco” ID 167-P Boumezzough A., Kahime K., Boussaa S. and Ouanaimi F. “Leishmaniasis in Morocco: still a topical question” ID 168-P Remondegui C. V., Cabrera C. H. A., Quintana M. G. “Seasonal fluctuation of Phlebotominae (Diptera: Psychodidae) in three Departments of an endemic area of Cutaneous Leishmaniasis in Jujuy province. A preliminary report” ID 169-P Bisetto Júnior A., Bisetto M., Dotto Ampessam S. M. “Spatial analyisis of the accurrence of leishmaniasis in Paraná, Brazil, to identify areas of risk” 15 ID 171-P Casanova C., Fernandes Shimabukuro P. H., Almeida Bressiani B., Fernanda Lazari T., Galati E. A. B. “Geographical distribution of Nyssomyia neivai (Pinto) and Nyssomyia intermedia (Lutz & Neiva) in the state of São Paulo, Brazil” ID 172-P Bergo E., Nascimento L., Camargo-Neves V. “First meeting of Lutzomyia longipalpis in rural areas related to outbreak of canine visceral leishmaniasis in the northwestern region of São Paulo, Brazil” ID 173-P Colla-Jacques F. E., Casanova C., Galvis Ovallos F. “Study of sand fly community in two rural endemic areas for American cutaneous leishmaniasis in the municipalities of Mogi Guaçu and Mogi Mirim, São Paulo State, Brazil” ID 174-P Saraiva L, Rugani J. M. N., Lima A.C.V. M. R., Reis A.S., Pereira A. A. S., Gontijo C.M.F., Andrade Filho J.D. “DNA detection of Leishmania sp. in female sand flies (Diptera: Psychodidae) in Belo Horizonte city and Sumidouro State Park, Minas Gerais – Brazil” ID 175-P Santini M. S., Utges M. E., Casas N., Salomon O. D. “Association Of Occurrence And Abundance Of Lutzomiya longipalpis with Environmental Variables At Different Spatial Scales: Micro And Macro-Scale” ID 176-P Waleed Al Salem, Badriah Al Otaibi, Louise Kelly-Hope, Karina Mondragon-Shem, Salah Balgonaeem, Maha Abdeladhim, Mohammed Al Zahrani, Amir Hassan, Jesus Valenzuela Alvaro Acosta-Serrano “A Public Health Approach to Leishmaniasis Control” INDEX BY TOPIC Vector Control 1-O, 2-O, 3-O, 4-O, 5-O, 6-O, 7-O, 8-O, 9-O, 10-O, 11-O, 12-O 1-P, 2-P, 3-P, 4-P, 5-P, 6-P, 7-P, 8-P, 9-P, 10-P, 11-P, 176-P Taxonomy & Phylogeny 13-O, 14-O, 15-O, 16-O, 17-O, 18-O, 19-O, 20-O, 21-O, 22-O, 23-O, 24-O 47-P, 48-P, 49-P, 50-P, 51-P, 52-P, 53-P, 54-P, 55-P, 56-P, 57-P, 58-P, 59-P, 60-P, 61-P, 62-P, 63-P, 64-P, 65-P, 66-P Ecology & Epidemiology 25-O, 26-O, 27-O, 28-O, 29-O, 30-O, 31-O, 32-O, 33-O, 34-O, 35-O, 36-O, 37-O, 38-O, 39-O, 40-O, 41-O, 42-O. 43-O, 44-O, 45-O 12-P, 13-P, 14-P, 15-P, 16-P, 17-P, 18-P, 19-P, 20-P, 21-P, 22-P, 23-P, 24-P, 25-P, 26-P, 27-P, 28-P, 29-P, 30-P, 31-P, 32-P, 33-P, 34-P, 35-P, 36-P, 37-P, 38-P, 39-P, 40-P, 41-P, 42-P, 43-P, 44-P, 45-P, 46-P, 67-P, 68-P, 69-P, 70-P, 71-P, 72-P, 73-P, 74-P, 75-P, 76-P, 77-P, 78-P, 79-P, 80-P, 81-P, 82-P, 83-P, 84-P, 85-P, 86-P, 87-P, 88-P, 89-P, 90-P, 91-P, 123-P, 124-P, 125-P, 126-P, 127-P, 128-P, 129-P, 130-P, 131-P, 132-P, 133-P, 134-P, 135-P, 136-P, 137-P, 138-P, 139-P, 140-P, 141-P, 142-P, 143-P, 144-P, 145-P, 146-P, 147-P, 148-P, 149-P, 150-P, 151-P, 152-P, 153-P, 154-P, 155-P, 156-P, 157-P, 158-P, 159-P, 160-P, 161-P, 162-P, 163-P, 164-P, 165-P, 166-P, 167-P, 168-P, 169-P, 170-P, 171-P, 172-P, Boletín de la Sociedad Entomológica Argentina N° 26 (1) 2015 16 173-P, 174-P, 175-P Vector Competence & Infections 46-O, 47-O, 49-O, 50-O, 51-O, 52-O 92-P, 92-P, 93-P, 94-P, 95-P, 96-P, 97-P, 98-P, 99-P, 100-P, 102-P, 102P, 103-P, 104-P, 105-P Leishmania – Sandfly /host Interactions 48-O, 53-O, 54-O, 55-O, 56-O, 57-O, 58-O, 59-O, 60-O 108-P, 109-P Genome & Genetics/ Virus infections 61-O, 62-O, 63-O, 64-O, 65-O, 66-O, 67-O, 68-O, 69-O 106-P, 107-P, 109-P, 110-P Physiology & Colonization 70-O, 71-O, 72-O, 73-O 111-P, 112-P, 113-P, 114-P, 115-P Other Topics 116-P, 117-P, 118-P, 119-P, 120-P, 121-P, 122-P INDEX BY AUTHOR (SURNAME) A Abassi Ibrahim 36-O, 34-P, 41-P, 162-P Abdeladhim Maha 8-O, 28-O, 112-O, 176-P Abdeliyev Ziyat 9-P Aboud Marium A 66-O Abras A 145-P Abreu Bruna Lacerda 136-P Abril Marcelo 44-P Acardi Soraya Alejandra 3-P, 45-P Acel Anagela Maria 26-P, 27-P, 28-P Acosta Luz A 26-O, 8-P, 63-P Acosta M Manteca 11-O Acosta-Serrano Alvaro 8-O, 28-O, 176-P Afonso Margarete Martins dos Santos 18-P, 19-P Afonso MO 33-P, 57-P, 111-P, 154-P Aguiar NO 151-P Aguilar E 27-O, 126-P, 127-P Aimakhanov Batirbek 9-P Akhavan AA 40-O Aklilu Esayas 37-O Alexandre Thiago Nunes 81-P Al-Hazza Ibraheem M 117-P Alkan C 39-O Al-Khalifa Mohammad S 117-P Almazán María Cristina 67-P Al Otaibi Badriah 112-O, 176-P Alrabba Fathi M 66-O Al Salem Waleed 28-O, 8-O, 176-P Alten B 21-O, 39-O, 157-P Alves Leucio Câmara 31-P Alves Maria de Lima 118-P, 119-P, 120-P, 146-P, 149-P, 156-P Alves-Pires C 111-P, 154-P Alves Veracilda Ribeiro 20-P, 22-P, 55-P Al Zahrani Mohamed 8-O, 28-O, 176-P Andrade-Filho José Dilermando 65-O, 13P, 25-P, 58-P, 60-P, 66-P, 72-P, 101-P, 128-P, 134-P, 136-P, 137-P, 139-P, 174-P Alzogaray Raul A 5-P Amini Esfanjani MH 61-P Ampessam Sônia Maria Dotto 169-P Añez Néstor 25-O Arandian MH 40-O Araujo Analia V 38-P, 83-P Archer Reginald 155-P Arcones Carolina 17-O, 22-O Arheart Kristopher L 155-P Arivillaga Jazzmin 162-P Arserim Suha K 4-O, 36-P, 37-P Atshabar Bakhit 9-P Austin James 1-O Aydın S 39-O B Balcioğlu I Cüneyt 37-P, 4-O Balgonaeem Salah 8-O, 176-P Balkew Meshesha 35-O, 36-O, 37-O, 38-O, 29-P, 34-P, 90-P Ballart C 145-P, 154-P Baneth Gad 73-P Banuls A-L 39-O Barata Iorlando da Rocha 92-P, 112-P Barazesh A 61-P Barbosa Diogo Brunno e Silva 123-P Bargues María Dolores 18-O Barhoumi W 3-O, 44-O, 132-P, 155-P Barresi Trey 2-O Barroso Paola A 67-P Basano Fabrizio Solari 12-O Basmadjián Yester 122-P, 141-P, 142-P, 150-P Bates MD 100-P Bates Paul A 50-O, 55-O, 100-P Batista Hebert Lima 78-P Becker-Fauser Ingeborg 105-P Beier John C 155-P Beitz Eric 115-P Bejarano Eduar Elías 26-O, 46-O, 51-O, 8P, 59-P, 63-P, 91-P Belen A 39-O Bell Melissa 46-P Ben Helel K 132-P Ben Salah Afif 49-P, 96-P Bénié Goze Bertin 32-O Benítez Ibalo P 38-P Bergo Eduardo 172-P Berrozpe Pablo 83-P Bertollo Denise Maria Bussoni 109-P Berzuna-Cruz Miriam 105-P Bettaieb Jihene 32-O, 96-P Bettiol Flávia Karolina Pereira Barreto 27P, 28-P Bevilaqua Claudia ML 16-P Bevilacqua Paula Dias 72-P, 60-P Bianchi R 68-O Bichaud L 44-O Bisetto Júnior Alceu 11-P, 169-P Bisetto Maurício 169-P Blanda M 82-P Bongiorrno G 45-O, 68-O Borda Carlos Edgardo 62-P, 121-P, Borges-Alves G 14-O Botelho Patrick Feitosa 54-P Boujaâma A 3-O Boumezzough Ali 106-P, 164-P, 165-P, 166-P, 167-P Boussaa Samia 106-P, 164-P, 165-P, 166-P, 167-P Brahim L 102-P Brandão-Filho Sinval Pinto 12-P, 30-P, 31P, 32-P, 152-P Brandão Roberto Carlos Feitosa 92-P, 112P N° 26 (1) 2015 Bray DP 14-O Brazil Reginaldo Peçanha 14-O, 60-P, 72P, 124-P Bressiani Bruna Almeida 171-P Brianti Emanuele 12-O Brilhante Andreia Fernandes 17-P Browers Marinês 2-P Burleigh Barbara 69-O Burruss Dylan 2-O, 3-O C Cabrera CHA 168-P Cabrera Olga Lucía 7-O, 9-O, 10-O Cáceres Abraham G 15-O Cadavid Gloria 73-O Cadena Horacio 26-O, 8-P, 63-P, 159-P Caligiuri Lorena G 3-P Calvopiña Manuel 162-P Calzada Jose E 35-P Camargo-Neves Vera 172-P Cameron Mary 46-P Campino L 33-P, 57-P, 111-P Campos Marliane Batista 112-P Canneva Bruno 122-P, 141-P, 142-P, 150P Caranha Lindemberg 89-P Cardoso A 140-P Cardoso Cristiane de Oliveira 17-P Carneiro Mariângela 135-P Carpenter S 50-O, 143-O Carter V 14-O Carvalho AK 97-P Carvalho Gustavo Mayr de Lima 25-P, 136-P, 139-P Carvalho Silvia María Santos 144-P Casanova Claudio 171-P, 173-P Casaril Aline Etelvina 84-P, 85-P, 88-P, 128-P Casertano Sergio A 58-P, 125-P Castilho Regiane de Cassia Hilário 109-P Castillo G 67-P Cavalcanti Milena de Paiva 32-P Cavia Regino 21-P Çetin Hüseyin 4-O Chacon M 27-O, 127-P, 147-P Chagas Anadeiva Portela 30-O Chalghaf Bilel 32-O Chanampa Mariana M 67-P Charrel RN 39-O, 44-O Chaves Sérgio Augusto de Miranda 18-P, 19-P Chavez Luis F 35-P Checa Rocío 22-O, 17-O Chemkhi Jomaa 49-P, 96-P Chelbi Ifhem 3-O, 44-O, 155-P Cherni Saifedine 3-O, 44-O, 155-P Chicharro Cármen 17-O, 22-O Boletín de la Sociedad Entomológica Argentina Chiluisa Orlando 162-P Chlebi Ifhem 72-O Chlif Sadok 32-O Chouihi E 132-P Ciufolini MG 68-O Cochero Suljey 46-O, 51-O, 91-P, Coelho Tháis Oliveira 6-P Colla-Jacques Fernanda Elisa 173-P Collins Frank H 62-O Contreras Angélica 159-P Copa Griselda N 67-P, 158-P Corbett CEP 97-P Cordeiro Jose LP 42-P Cordovez Juan Manuel 153-P Corrêa Arlene Gonçalves 71-O Cortés Luis 51-O, 91-P Cosenza M 23-O Costa Carlos Henrique Nery 10-P, 123-P Costa Dorcas L 10-P Costa Filho Pedro Catarino 143-P Costa Pietra Lemos 12-P, 30-P Costa Simone M 18-P Courtenay O 14-O, 170-P Cristóvão JM 33-P, 57-P, 111-P Cruz Israel 17-O, 22-O Cruz Susana 22-O Cumbrera Anayansi Valderrama 65-O Cupollilo Elisa 116-P Cutolo Andre Antonio 104-P D Dachraoui K 44-O, 132-P da Costa Ailton Junior Antunes 74-P, 75-P da Costa Simone Miranda 19-P, 42-P da Fonseca Claudio Ruy V 55-P D'Agostino R 23-O Dagur Pradeep K 56-O Damazo Amilcar Sabino 1-P Dantas-Torres Filipe 12-P, 30-P,31-P, 32-P, 152-P, 12-O Darabi H 40-O Darvishi M 40-O Da Paixão Fabiana Rodrigues 75-P da Silva Ana de Nazaré Martins 76-P da Silva Andrea Martins 69-O da Silva Danyele Franca 134-P da Silva Fábio Márcio Medeiros 54-P, 92-P, 112P da Silva Fernando José 12-P, 30-P, 152-P da Silva Jalthon C 10-P da Silva José Hilário Tavares 146-P, 149-P, 156-P da Silva Junior João Batista 118-P, 119-P, 156-P da Silva Kamila Gaudêncio 31-P, 32-P, 152-P da Silva Luiz Osvaldo Rodrigues 89-P 17 Da Silva Maria Elisabel Tavares 77-P da Silva Mariana Dantas 104-P da Silva Shara Regina 74-P da Silva Túllio Romão R 94-P Das Pradeep 6-O Davidson Silas 7-P De Almeida Andréa Sobral 135-P de Almeida Paulo Silva 29-O, 143-P de Almeida Romoaldo Martins 143-P de Almeida Tiago Parreiras 27-P, 28-P de Andrade Andrey José 13-O, 29-O, 6-P, 14-P, 50-P, 52-P, 56-P de Araújo Magnólia Florêncio Fernandes 119-P de Araújo Patrick Thiago Carvalho 76-P de Assis Mauro Diego Gobira Guimarães 94-P de Avelar Daniel Moreira 74-P de Carvalho Bruno M 18-P, 19-P, 40-P de Carvalho Gílcia Aparecida 31-P de Carvalho Maria do Socorro Laurentino 29-O de Carvalho RMJS 140-P de Figueiredo Helen Rezende 88-P, 128-P De Lazzari Elisa 1-O de Medeiros Jansen Fernandes 98-P, 129-P, de Melo Fábio Lopes 12-P Demir Samiye 4-O de Moraes Sinara Cristina 80-P, 81-P de Morais Rayana Carla Silva 32-P de Oliveira Alexandra Gutierrez 84-P, 85-P, 88-P, 101-P, 128-P de Oliveira Everton Falcão 88-P Depaquit Jerome 16-O, 18-O, 20-O, 21-O, 24-O, 64-O de Paula Marcia Bicudo 143-P de Pita Pereira Daniela 19-P Derbali Mohamed 3-O, 44-O, 155-P Derrick Tamsyn 55-O de Santana Antônio Luis Ferreira 19-P de Souza Emerson Marques 27-P, 28-P de Sousa Gilberto César 30-O de Souza Jailson Ferreira 17-P de Souza Marcos Barbosa 110-P de Souza, Nataly A 48-P de Souza Wagner Fernnades 84-P, 85-P Deuster Katrin 12-O Dey Ranadhir 56-O Dias Edelberto Santos 1-P, 74-P, 75-P, 79-P, 93-P, 135-P Dias Quezia Nunes Peres 110-P Dias TA 25-P Diaz-Albiter Hector M 67-O Diaz Mery 86-P, 87-P Di-Blasi Tatiana 54-O, 59-O Dilger E 14-O, 170-P Dillon Roderick J 55-O, 67-O N° 26 (1) 2015 Di Muccio T 45-O Dinesh Diwakar Singh 6-O Direni-Mancini JM 65-P, 163-P Donato Lucas Edel 16-P do Nascimento Thais Brito 81-P Dorval Maria Elizabeth Moraes Cavalheiros 101-P dos Santos Ana Paula de Azevedo 98-P dos Santos Antonio Pereira 89-P Dos Santos Carlos Fernando Rocha 15-P, 77-P dos Santos Eric Fabricio Marialva 129-P dos Santos Fabio Alexandre Leal 27-P dos Santos Klieton Maciel 101-P dos Santos Lucas Heber Mariano 81-P dos Santos Nathanna Progênio 17-P dos Samtos Paulo Raimundo Barbosa 144P dos Santos Thiago Vasconcelos 48-P, 54-P, 76-P, 92-P, 112-P dos Santos Yago Costa Vasconcelos 76-P Driss Mahdi 96-P Duarte Didier Tirado 159-P Duffy Darragh 57-O Duncan Robert 56-O D'Urso V 45-O Dvořák Vít 19-O, 20-O, 21-O, 39-O E Echchakery Mohamed 106-P, 164-P, Echeverría Jorge Espinoza 86-P, 87-P Eguchi Gabriel Utida 84-P, 85-P, 88-P Elnaiem Dia-Eldin A 31-O, 66-O Emrich Scott J 62-O Enriquez Sandra 162-P Ergunay K 39-O Ermis V Ozge 37-P Escobedo-Ortegón Javier 105-P Espada Liz 64-P Estrada Luis G 26-O, 8-P, 63-P F Fadul Mônica 76-P Faiman Roy 39-P Fares W 44-O Faria Maiara d 67-O Farias LHS108-P Farias Teresinha C 10-P Faustino Maria Aparecida da Gloria 31-P Fernándes Carlos Eurico 84-P, 85-P Fernandes Magda Freitas 101-P Fernandes Wedson Desidério 101-P, 124-P Fernández María Soledad 21-P, 43-P, 160-P Fernández Roberto 64-P Fernández Wagner de Souza 88-P Ferrari Júnior Antonio Carlos 101-P Ferreira Fernando 70-P, 71-P Boletín de la Sociedad Entomológica Argentina Ferreira Gabriel Eduardo Melim 116-P Ferreira Jônatas Barbosa Cavalcante 50-P Ferreira Maria 133-P Ferro Cristina 153-P, 161-P Ferrolho J 111-P Figueiredo Juliana Kindler 109-P Fiuza Vanessa de Oliveira Pires 74-P Fortes-Dias Consuelo Latorre 74-P, 79-P Fortier Daniel Costa 123-P Fortuna C 68-O França-Silva João Carlos 75-P Francesquini FC 97-P Franco Luciano 133-P Freire Maíra Posteraro 20-P, 94-P Freire Renato Cesar Melo 149-P Freitas FB 33-P Freitas RA 151-P Freitas Vanesa C 48-O Fuenzalida A Denise 65-P, 125-P, 163-P Fuller Douglas O 155-P Fux Blima 13-P Fuya Patricia 153P, 161-P G Galati Eunice Aparecida Bianchi 13-O, 17P, 48-P, 51-P, 52-P, 56-P, 70-P, 71-P, 88-P, 101-P, 104-P, 143-P, 171-P Galindo B 147-P Gállego M 145-P, 154-P Gálvez Rosa 17-O, 22-O Galucci Estela 70-P, 71-P Gama Renata Antonaci 149-P Gamarra Roberto M 124-P Garcez Lourdes María 30-O Garciá Ana Lineth 86P, 87-P Garlapati Rajesh B 2-O Garrido Renata 18-P Gaudêncio Kamila 30-P Gebre-Michael Teshome 33-O, 34-O, 35O, 36-O, 37-O, 38-O, 29-P, 34-P, 39-P, 41P, 90-P Gebresellassie Araya 33-O, 34-O, 36-O, 39-P, 41-P, 90-P Gendernalik Alex 9-P Genta Fernando A 67-O Gharbi M 132-P Gil José 67-P, 158-P Giordano Cristina 19-P Giraldo-Calderón Gloria I 62-O, 63-O Giraud Emile 55-O Godoy Rodrigo Espíndola 48-P, 54-P Gomes J. ID 111-P Gomes Suellem Petilim 88-P Gómez Bravo Andrea 44-P Gómez-Bustamante Marinely 86-P, 87-P Gómez Hernando 8-P Gonçalves Raquel 30-O 18 Gonçalves Rodrigo Gurgel 6-P Gontijo Célia Maria Ferreira 25-P, 66-P, 95P, 134-P, 139-P, 174-P González-Britez Nilsa 11-P, 133-P González Camila 153-P, 161-P González E 145-P González Estela 99-P, 130-P, 131-P Goulart Thais Marchi 71-O, 113-P, 114-P Gould Ignacio Tomás 5-P, 43-P, 44-P, 125-P Gradoni Luigi 73-P, 45-O Gramiccia Marina 73-P, 45-O, 68-O Grant William E 5-O Graser Carina 18-P Guerbouj Souheila 49-P Guimarães Ricardo 30-O Guimarães Vanessa Cristina Fitipaldi Veloso 12-P, 30-P, 144-P Guizani Ikram 49-P Gunay 39-O Gurgel-Gonçalves Rodrigo 29-O, 14-P, 50P, H Habboul Z 132-P Hadhri MH 132-P Hailu Asrat 19-O, 33-O, 34-O, 29-P, 34-P, 35-O, 36-O, 37-O, 38-O, 39-P, 41-P, 73-P, 90-P Halada Petr 20-O Hamida Nabil Belhadj 58-O Hamilton J Gordon C 14-O, 72-O Harrabi Mariem 32-O Hasan Milena 57-O Hashiguchi Yoshihisa 15-O Hassan Amir 112-O, 176-P Hassan Faizan 6-O Hawksworth Mallory 63-O Hermes Sandra Cristina Negreli Moreira 29-O Hernández Sonia 130-P, 131-P Hiramoto Roberto Mitsuyoshi 109-P Hlavacova J 60-O Hlavačková Kristýna 19-O, 20-O I Ilango Kandan 47-P Infran Jucelei de Oliveira Moura 84-P, 85P, 88-P, 128-P Intile S 82-P, 23-O Ipe Ipe M 43-O Ishikawa Edna Aoba Yassui 76-P J Jafari R 40-O Jiménez Maribel 4-P, 99-P, 130-P, 131-P, 145-P Joshi Amritanshu 56-O Juan Laura W 5-P N° 26 (1) 2015 K Kahime Kholoud 106-P, 164-P, 165-P, 167-P Kamhawi Shaden 53-O, 56-O, 115-P Karakus Mehmet 4-O, 36-P Kasap Ozge Erisoz 21-O, 39-O, 157-P Kassahun Aysheshm 19-O Katakura Ken 15-O Kato Hirotomo 15-O Kelly-Hope Louise 8-O, 28-O, 176-P Khajeian M. 40-O Khalid Noteila M 66-O Khoury C 45-O, 68-O Kindlová Michaela 73-P Kirstein Oscar 33-O, 34-O, 36-O, 39-P, 41P, 90-P, 162-P Kobylinski Kevin 7-P Košťálová Tatiana 73-P Kowalewski Martín 148-P Kozhahmetova Madina 9-P Kratochvílová Tereza 73-P Kumar Vijay 6-O Kwakye-Nuako G 100-P L Lacerda Helio Márcio Nunes 78-P Lagarmilla Patricia 122-P, 141-P, 142-P, 150-P Lal Das Murari 1-O Lal Dinesh 43-O Lamballerie X de 44-O Lana Rosana Silva 79-P Lara-Silva Fabiana de Oliveira 74-P, 75-P, 79-P La Russa F 82-P Lastre Natalia 59-P Laurenti MD 97-P Lavitschka Cecilia Oliveira 104-P Lawson Daniel 62-O Lazari Thaís Fernanda 171-P Leão Edna De Freitas 112-P Léger Nicole 16-O, 24-O Lehrter Veronique 20-O Leite Keily Cristiny Azevedo 78-P Lelli R 82-P Lemma Wossenseged 33-O, 35-O, 29-P, 34-P Levay Ana Paula Silva 101-P Libri Valentina 57-O Lima Ana Cristina Vianna Mariano da Rocha 74-P, 75-P, 174-P Lima Fernanda Pinheiro 13-P Lima Jan108-P Lima José Aprígio Nunes 112-P Lima Magliones Carneiro 15-P, 77-P Limam S 132-P Lisi O 45-O Boletín de la Sociedad Entomológica Argentina Lizarralde de Grosso MS 163-P Lopes Josiane Valadão 75-P Lopes Tamy Elicia da Silva 118-P, 119-P, 146-P, 149-P, 156-P Lopez Marla 153-P Lorini Maria Lucia 40-P Louzir Hechmi 57-O, 58-O Lozano Alejandra 122-P, 141-P, 142-P, 150-P Lubbad MY 117-P Lugo Ligia 153-P, 161-P Luna R 127-P LL Llanes-Acevedo Ivonne Pamela 17-O, 22-O Llanos A 27-O M Macena Ana Lúcia Moreira 15-P, 77-P Machado Caroline Amaral 22-P, 116-P Machado Vicente Estevam 71-O, 113-P, 114-P Machado-Coelho George Luiz Lins 74-P Maia C 33-P, 57-P, 111-P Magalhães Mônica de Avelar Figueiredo Mafra 18-P Mandal Goutam 115-P Mandal Srotoswoti 115-P Marceló Catalina 9-O Marchi A 68-O Marchini Fabricio K 59-O, 69-O Marco JD 158-P Margonari Carina 13-P Maroli M 45-O Mas-Coma Santiago 18-O Massoudi Nabil 132-P Mastrángelo AV 11-O Martín-Martín Inés 4-P, 99-P, Martín Ohiane 17-O, 22-O, 55-O Martín-Sánchez J 154-P Martínez Lily Paola 46-O, 51-O, 91-P Martínez Mariela F 148-P Martínez Nidia 133-P Martins Juliana 79-P Mateus Nathália Lopes Fontoura 88-P Mathis Alexander 20-O Matthew Albert 57-O Mazloumi Gavagni A 61-P McCarthy Christina B 3-P McCoy John Philip 56-O McDowell Mary Ann 61-O, 63-O, Meddeb-Garnaoui Amel 57-O, 58-O Medley Graham 170-P Meira Antônio de Medeiros 110-P Melo Alan Lane 80-P Memish Ziad 28-O Mendes Adauto Rodrigues 128-P 19 Mendes Ileane Gurgel 119-P Mendonça Ivete L 10-P Mendoza J 127-P Menegatti Jaqueline Aparecida 29-O Meneghello Brígida Helena da Silva 109-P Meneses Claudio 115-P, 70-O, 56-O Menezes Filho Antônio 10-P Menezes Júlia Alves 13-P Menz Ingrid 104-P Mermer Ahmet 37-P Mhadhbi M 132-P Michalsky Érika Monteiro 74-P, 75-P, 79-P, 93P Mierez Mirta Liliana 62-P Miguelañez Silvia 17-O, 22-O Miranda Débora Elienai de Oliveira 31-P, 32-P, 152-P Miró Guadalupe 17-O, 22-O Miyazaki Rosina Djunko 26-P, 27-P, 28-P, 93-P Molina Juan 11-O, 5-P Molina Ricardo 4-P, 99-P, 130-P, 131-P, 145-P Molla Leticia María 104-P Monaco Neiva Zandonaide 88-P Moncaz Aviad 33-O, 34-O, 36-O, 90-P Mondragon-Shem Karina 28-O, 46-P, 176P Montesino Ana 51-O, 91-P Montoya Ana 17-O, 22-O Moraes Carolina d. 67-O Morais María Helena Franco 135-P Morais Mayron Henrique Gomes 13-P Morales E 70-O Morilla F 154-P Moreno Eduardo Stramandinoli 20-P Moreno H Claudia Ximena 73-O Mosqueda Luis Armando 62-P Moura Abilene Cristina de Arruda 32-P, 152-P Mukhopadhyay Rita 115-P Műller Ingrid 55-O Murgas Ingrid 65-O N Nakashi Hira L 56-O Naouar Ikbel 57-O, 58-O Nascimento Lucimar 172-P Nascimento Luis Otávio Cordeiro 20-P Nasser Julio R 67-P, 158-P Negrão Fábio Juliano 101-P Neto Grimoaldo Braga Da Rocha 15-P, 77P Nogueira Fabio dos Santos 104-P Novo Shênia Patrícia Corrêa 110-P Nowling RJ 63-O Nzelu Chukwunonso Onyemaechi 15-O N° 26 (1) 2015 O Obara Marcos Takashi 6-P Oliveira AFJ 151-P Oliveira Alessandra G 124-P Oliveira Ana Flávia de Morais 78-P Oliveira DMS 108-P OliveiraEverton Falcão 128-P Oliveira Fabiano 53-O, 70-O Ölgen M. Kirami 4-O Oreste F 67-P Orta Jose 115-P Ortega Edgar 26-O, 8-P, 63-P Ortiz Washington Benitez 162-P Oscherov EB 38-P Oshiro Elisa Teruya 84-P, 85-P, 88-P, 101-P, 128-P Osuguku E 27-O Otranto Domenico 12-O, 152-P Ouanaimi Fouad 106-P, 164-P, 165-P, 166-P, 167-P Ovallos Fredy Galvis 70-P, 71-P, 104-P, 173P Özbel Y 4-O, 39-O, 36-P, 37-P Özkul A 39-O P Pacheco R 27-O, 102-P, 103-P, 126-P, 127P, 147-P Pandey Shubham 43-O Paranhos Filho Antonio C 124-P Pardo Raúl H I 7-O, 9-O, 10-O Parente Daniela M 10-P Parrado Rudy 86-P, 87-P Parras Matías Ariel 23-P, 24-P, 68-P, 69-P, 125-P ParreiraR. 57-P Passero LF 97-P Paternina Luis 46-O Paternina Margaret 46-O Paz Gustavo Fontes 139-P Pech-May Angélica 105-P Pereira AAS 174-P Pereira Araujo T 102-P Pereira-Chioccola 104-P Pereira Júnior Antonio Marques 98-P, 129-P Pérez Adriana Alicia 11-O, 43-P Pérez-Doria Alveiro 46-O, 51-O, 59-P, 91-P Perez JE 27-O, 103-P, 126-P, 127-P PessanhaJosé Eduardo Marques 74-P Pessoa Felipe Arley Costa 98-P, 129-P, 151P Pessoa Grasielle CD 16-P Pesson B 154-P Pflüger Valentn 20-O Philips-Singh Doris 43-O Piazza M 23-O Picado Alberto 1-O Boletín de la Sociedad Entomológica Argentina Pinheiro Marco Paulo Gomes 118-P, 119-P, 120-P, 146-P, 149-P, 156-P Pinheiro Maria Paula de Melo Pereira 120-P Pinheiro Maria Sueli Barros 92-P, 112-P Pinto Israel de Souza 116-P Pinto Mara Cristina 71-O, 113-P, 114-P Pimenta Paulo PFP 48-O, 52-O Pitaluga Andrè N 54-O, 69-O Pita-Pereira D 102-P Pires Ana Clara Araújo Machado 52-O Pitti R 23-O Poché David M 5-O, 9-P Poche Richard 2-O, 3-O, 9-P Polyakova L 3-O, 9-P Ponte Cesar do Santo 110-P Portillo Nilda 133-P Portús M 145-P, 154-P Posada Laura 46-P Posada-López Laura 73-O, 63-P, 159-P Pozo Catherine 162-P Prudhomme Jorian 20-O Pruzinova K 60-O Q Quaresma Patrícia Flávia 25-P, 60-P, 72-P, 95-P Qualls Whitnay A 155-P Quintana María Gabriela 11-P, 65-P, 163-P, 168-P Quispe Anita 27-O, 103-P, 126-P, 147-P Quispe C 147-P Quispe D 27-O, 126-P Quispe-Florez MM 102-P, 103-P Quispe M 27-O Quispe-Ricalde MA 103-P, 147-P R Rado D 27-O, 102-P, 103-P, 126-P, 127-P, 147-P Rahola Nil 53-P Ramalho Miriele 152-P Ramalho-Ortigao José Marcelo 59-O, 115P Randrianambinintsoa Fano José 16-O Rangel Elizabeth F 16-P, 18-P, 19-P, 40-P, 42-P, 48-P, 80-P, 89-P, 102-P, 126-P Raizer Josué 29-O Ravel C 145-P Rea María Josefa Felisa 62-P, 121-P Ready PD 47-O Reale S 23-O, 82-P Rebai-Kammoun Wafa 57-O, 58-O Rebolar-Téllez Eduardo A 105-P Remondegui CV 168-P Rêgo Felipe Dutra , 25-P, 94-P, 95-P, 116-P, 134-P, 138-P, 139-P Rego Taiana Amâncio da Costa 19-P 20 Reis AS 66-P, 174-P Reithinger Richard 86-P Remoli ME 68-O Rhodes C 70-O Ribeiro Ana Lucia Maria 1-P, 26-P, 27-P, 28-P, 93-P Ribeiro Carlos CB 10-P Ribeiro Leticia Moraes 128-P Rigg Chystrie 35-P Ríos Cristina 122-P, 141-P, 142-P, 150-P Rocha Anárya Teresa de Freitas 123-P Rodarte Cláudia Cavalcante De Matos 77P Rodrigues Eduardo Henrique Gomes 12-P Rodrigues Jorge Senatore Vargas 1-P, 26-P, 27-P, 28-P Rodrigues Moreno de Souza 129-P Rodríguez Jorge Luis 51-O Rodríguez-Jiménez Jorge 91-P Rocha Douglas de Almeida 6-P, 50-P Rogers Matthew E 55-O Rohoušová Iva 73-P Rojas Agustina 25-O Rojas de Arias Antonieta 11-P Romero Luis Roberto 51-O Romero Selva 122-P, 141-P, 142-P, 150-P Romero Vanesa 162-P Romero-Ricardo Luis 59-P, 91-P Rosa Juan Ramón 23-P, 24-P, 38-P, 68-P, 69-P Routray Parimita 46-P Rowland Mark 1-O Rowland Tobin 7-P Rowton Edgar 7-P Ruiz-Lopez Freddy 159-P Rugani Jerônimo Marteleto Nunes 25-P, 95P, 174-P S Sábio Priscila Bassan 51-P, 52-P, 56-P Sabroza Paulo Chagastelles 135-P Sadlova Jovana 49-O, 50-O Saeidi Z 61-P Salah Afif B. 32-O, 57-O, 58-O Saldaña Azael 35-P Salomón Oscar Daniel 11-O, 3-P, 5-P, 11-P, 21-P, 23-P, 24-P, 38-P, 43-P, 44-P, 45-P, 58P, 65-P, 68-P, 69-P, 83-P, 125-P, 148-P, 160-P, 163-P, 175-P Samantaray Sachidananda 2-O Sánchez-García Laura 105-P Sandoval Adolfo Enrique 45-P, 225-P Sanguinette Cristiani de Castilho 134-P, 136-P Santamaría Erika 7-O, 9-O, 10-O Sant'Anna Mauricio R 67-O Santini María Soledad 11-O, 3-P, 11-P, 21- N° 26 (1) 2015 P, 45-P, 148-P, 160-P, 175-P Santos Fabricio K. M. 16-P, 89-P Santos Luciene Aranha da Silva 112-P Santos Mirella Ferreira da Cunha 124-P, 128-P Santos Nathália Queiroz 76-P Santos TV 108-P Santos Walter Souza 30-O Saracho-Bottero MN 163-P Saraiva Lara 66-P, 136-P, 174-P Satragno Dinora 122-P, 141-P, 142-P, 150-P Scholte Ronaldo Guilherme Carvalho 95-P Sciamarelli Alan 29-O Sebastien Boyer 16-O Seblova-Hrobarikova Veronika 49-O, 50-O Secundino Nágila FC 48-O, 52-O Sequeira Cirino 141-P, 142-P, 150-P Ser Önder 4-O Serra e Meira Paula Cavalcante Lamy 136-P, 137-P Sevá Anaiá da Paixão 70-P, 71-P Severini F 45-O Shahbazi F 40-O Shaw Jeffrey 41-O Shenker Moshe 33-O Shimabukuro Paloma Helena Fernandes 13-O, 20-P, 22-P, 55-P, 94-P, 95-P, 116-P, 171-P Silva AM 54-O Silva BJM 108-P Silva EO 108-P Silva João Carlos França 79-P Silva Júlia dos Santos 19-P, 89-P Silva Maiara Alves 74-P, 75-P Silva Marcel Miranda de Medeiros 146-P, 149-P, 156-P Silva Rafaella A 16-P Silva Simone Melo 123-P Silveira Fernando Tobias 92-P, 112-P Silveira FT 97-P, 108-P Šíma Michal 73-P Singh Mutum Ingobi 2-O Singh NS ID 43-O Soares Daniela Cristina 30-O Soares Márcia Maria Costa Nunes 109-P Soares Maria Regiane Araújo 123-P Soares Rodrigo Martins 70-P, 71-P Soleimani H 40-O Soto M 103-P, 127-P Sousa Alessandra Mara 13-P Sousa Lindemberg Caranha 16-P, 48-P Souza Daiana Alovisi 84-P, 85-P, 88-P Souza Cristian Ferreira 60-P, 72-P Souza GD 140-P Souza Mielle Barbosa 60-P Stanneck Dorothee 12-O Steinhorst Iris Ingrid 45-P Boletín de la Sociedad Entomológica Argentina Stell Frederick 64-P Sttump Rodolfo German Antonelli Vidal 134-P Suar Mrutyunjay 46-P Suau Hamide Aslan 55-O, 115-P Sumová Petra 73-P Supparo Eduardo 141-P Szelag Enrique Alejandro 23-P, 24-P, 68-P, 69-P T Tadros Samuel 63-O Tanure Aline 134-P, 139-P, 139-P Tartaglino Lilian 3-P Tavares Mara Garcia 65-O Tekie Habte 35-O, 36-O, 37-O, 29-P, 34-P, 90-P Teles Carolina Bioni Garcia 98-P Telleria Erich Loza 59-O Tempone Antonio Jorge 54-O, 59-O, 69-O Thies Sirlei Franck 1-P, 2-P, 27-P, 93-P Thomaz-Soccol Vanete 11-P Tinoco B 54-O Toccas F 27-O, 127-P, 147-P Tolezano José Eduardo 109-P Tomokane TV 97-P Tonelli Gabriel Barbosa 95-P, 134-P, 139-P Torales Martha 133-P Torina A 82-P Torina S 23-O Tort Cecilia 122-P, 141-P, 142-P, 150-P Toumi Amine 32-O, 57-O, 58-O Töz Seray 4-O Traub-Csekö Yara María 54-O, 59-O, 69-O Tripet Frederic 66-O Trueba Gabriel 64-O U Uribe Sandra I 8P, 26-O, 63-P, 73-O Utgés María Eugenia 11-O, 83-P, 125-P, 148-P, 175-P V Vaca Franklin 162-P Vaccalluzzo V 45-O Valderrama Anayansi 35-P Vale Mariana Moncassim 40-P Valencia B 27-O Valenzuela Jesus 8-O, 28-O, 53-O, 56-O, 70-O, 112-O, 115-P Valladares B 102-P, 103-P, 126-P, 127-P, 147-P, 176-P Vargas Jorge Senatore Rodrigues 61-P, 93-P Vásquez Gisella 64-P Vega Celeste 11-P Vela Gabriela 64-O Vélez Iván Darío 26-O, 8-P, 63-P, 159-P 21 Vélez-Mira Andrès 26-O, 8-P, 63-P, 159-P Verbel-Vergara Daniel 46-O Vergara José G 51-O, 91P Verger Lorenzo 122-P, 141-P, 142-P, 150-P Vianna Elisa Neves 135-P Vidal Joaquim 17-P Viera Ana 122-P, 141-P, 142-P, 150-P Vilela M 126-P Villacrés Ernesto 162-P Villarquide ML 160-P Vincent Robert 16-O Vitale Edgardo 122-P, 141-P, 142-P, 150-P Vitale F 23-O, 82-P Vivero Rafael J 26-O, 73-O, 8-P, 63-P Volf Petr 19-O, 20-O, 21-O, 39-O, 49-O, 50-O, 60-O, 73-P Votýpka Jan 19-O, 21-O, 39-O, 49-O, 60-O W Wadsworth Marinha 63-O Wang Hsiao-Hsuan 5-O Warburg Alon 19-O, 33-O, 34-O, 35-O, 36O, 37-O, 38-O, 42-O, 49-O, 29-P, 34-P, 39P, 41-P, 73-P, 90-P, 162-P Ximenes Maria de Fátima Freire de Melo 118-P, 119-P, 120-P, 146-P, 149-P, 156-P Y Yadon Zaida E 11-P Yaghoobi-Ershadi M. Reza 40-O Yared Solomon 33-O, 38-O Yazidi Rihab 96-P Yépez José Yancarlos 25-O Yilmaz Bahtiyar 4-O Z Zacarias Danielle A 10-P Zanini Volmir 20-P Zaffora G 23-O Zapata Sonia 20-O, 64-O Zenobio Ana Paula Lusardo de Almeida 136-P Zerba Eduardo N 5-P Zhioua Elyes 3-O, 72-O, 44-O, 132-P, 155-P Zoghlami Z 132-P Zorrilla Victor 64-P Zwetsch Adriana 48-P ABSTRACT BOOK ROUND TABLES & ORAL SESSIONS ISOPS VIII Puerto Iguazú Argentina N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 1-O Vector Control ID 2-O Vector Control EVALUATION OF DIFFERENT NETS AGAINST PHLEBOTOMUS ARGENTIPES, THE VECTOR OF VISCERAL LEISHMANIASIS IN NEPAL MULTIPLE FIPRONIL DOSES IN DAIRY CATTLE FOR VECTOR CONTROL IN BIHAR, INDIA: RESIDUE STUDY AND EFFICACY DETERMINATION AGAINST THE SAND FLY PHLEBOTOMUS ARGENTIPES Albert Picado; Murari Lal Das; Mark Rowland; James Austin; Elisa De Lazzari B a rc e l o n a C e n t re f o r I n t e r n a t i o n a l H e a l t h R e s e a rc h ( C R E S I B ) . [email protected] Keywords: Phlebotomus argentipes, insecticide treated nets, mesh size, cross-over trial 23 Rajesh B Garlapati1, Sachidananda Samantaray2, Mutum Ingobi Singh1, Trey Barresi1, Dylan Burruss1 and Richard Poche1 1 Genesis Laboratories, Wellington, Colorado, USA. 2Bihar Veterinary College, Patna, Bihar, India. [email protected] Keywords: Fipronil, cow, residue, sand fly In the Indian subcontinent, Leishmania donovani, the parasite causing visceral leishmaniasis (VL) is transmitted by the sand fly vector Phlebotomus argentipes. Long lasting insecticide treated nets (LN) have been postulated as alternative or complement to Indoor Residual Spraying but there are few field studies evaluating the entomological efficacy of different nets against this vector. We conducted two crossover trials in a VL endemic area in Nepal to compare the barrier effect of (1) LN with different mesh sizes (156 holes/inch2 vs 625 holes/inch2) and (2) alpha-cypermethrin treated LN and untreated nets having the same mesh size (156 holes/inch2). Each crossover trial had two arms consisting of a sequence of two different nets for 8 nights. We used 10 cattle sheds per trial as unit of analysis and a cow placed under the net as bait. CDC light traps placed inside the nets were used to evaluate the number of P. argentipes crossing the net barrier. Negative binomial generalized estimating equation (GEE) population-averaged models adjusted by night and sequence were used to estimate the barrier effect of the different nets. The crossover trials conducted in a rural village in Morang district (South-eastern Nepal) demonstrated that reducing the size of the holes in treated nets (625 holes/inch2) increased the barrier effect of LN by 77% (95% confidence interval (CI): 56% - 88%) compared with treated nets with larger holes (156 holes/inch2). Treating nets with alpha-cypermethrin reduced the number of P. argentipes captured inside the nets by 77% (95% CI: 27% - 93%) compared with untreated nets. The effect of finer mesh LN on VL prevention needs to be tested in a randomized controlled trial. Visceral Leishmaniasis is a protozoan disease prevalent in India. VL is a severe form of Leishmaniasis which is transmitted by sand flies. Phlebotomus argentipes is the only available vector for VL transmission in India. Vector control plays an important role in VL management. Cows are an important blood meal source for P. argentipes. A study was initiated to assess sand fly mortality in response to fipronil oral dosing of dairy cows. Twenty hybrid cows were selected and four different doses (0.25 mg/Kg body weight, 0.5 mg/Kg body weight, 1.0 mg/Kg body weight and 1.5mg/Kg body weight) of fipronil were given at 8 week intervals for six months, totaling three separate applications. Sand fly efficacy and fecundity were evaluated on Day 1, 4, 7, 14, 21, 28, 35, 42 and 49 after treatment. Fipronil residues in milk were assessed for the first 14 days after treatment. Plasma residues were assessed on Day 1, 4, 7, 14, 21 and 28 after treatment during three applications. Milk production, cow health and environmental conditions were monitored during the study. Higher doses of fipronil (1.0 mg/Kg body weight and 1.5 mg/Kg body weight) had high residues in milk and longer sand fly efficacy after treatment. The efficacy of the lowest dose (0.25 mg/kg body weight) was found up to 14 to 21 days after treatment. Fipronil did not have a significant effect on sand fly fecundity. Metabolism of cows played an important role in milk and plasma residues during three applications. The study results suggest that the use of fipronil as a feed through substance can be incorporated into Visceral Leishmaniasis management programs. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 3-O Vector Control ID 4-O Vector Control LABORATORY AND FIELD EVALUATION OF RODENT BAIT TREATED WITH FIPRONIL FOR FEED THROUGH AND SYSTEMIC CONTROL OF PHLEBOTOMUS PAPATASI N AT U R A L I N F E C T I O N A N D I N S E C T I C I D E SUSCEPTIBILITY STATUS OF WILD CAUGHT SAND F L I E S I N R U R A L A R E A S O F A N TA LYA , MEDITERRANEAN REGION OF TURKEY M Derbali1, L Polyakova2, A Boujaâma1, D Burruss2, S Cherni1, W Barhoumi1, I Chelbi1, R Poch2, E Zhioua1 Mehmet Karakuş1; Samiye Demir2; Hüseyin Çetin3; Suha Kenan Arserim4; Önder Ser5; Seray Töz6; I Cüneyt Balcioğlu7; M Kirami Ölgen8; Bahtiyar Yilmaz2; Yusuf Özbel6 1 Institut Pasteur de Tunis, Laboratory of vector Ecology, 13 Place Pasteur, BP 74, 1002 Tunis, Tunisia. 2Genesis Laboratory Inc., 10122 NE Frontage Road, Wellington, Co 80549, USA. [email protected] Keywords: Phlebotomus papatasi, Meriones shawi, zoonotic cutaneous leishmaniasis, systemic and feed through insecticide The sand fly Phlebotomus papatasi is the main vector of Leishmania major, etiologic agent of zoonotic cutaneous leishmaniasis (ZCL), which is endemic in North Africa, the Middle East, and Asia. In North Africa, Meriones shawi is one the main reservoir host of L. major. Phlebotomus papatasi populations are maintained in borrowing rodents such as M. shawi. Three fipronil-treated rodent baits were evaluated for systemic and feed through insecticidal activity against P. papatasi feeding on M. shawi. Through blood feeding bioassays, mortality rates of females P. papatasi increased with the concentration of fipronil in the rodent bait varying from 0.001% to 0.005%. In the laboratory, more than 90.0% of P. papatasi were killed within 48 hours after blood feeding on the desert's jirds, Meriones shawi, treated up to 29 days prior with a single application of fipronil at a concentration of 0.001%, 0.0025% and 0.005%. Through larval bioassays, mortality rates of larvae that have fed on faeces of treated bait for M. shawi increase with the concentrations of fipronil. Faeces of orally-treated Meriones were significantly toxic to larvae for 5 weeks with a concentration of 0.005%. In the field, application of treated bait resulted in 80.0% reduction in the populations of P. papatasi up to 6 weeks after a single application of fipronil at a concentration of 0.005%. This is the first study to demonstrate field efficacy of fipronil-treated rodent baits for P. papatasi control and the first study to evaluate this approach in M. shawi, a principal ZCL reservoir host. These results suggest that fipronil-treated rodent baits can be used to effectively reduce the populations of P. papatasi associated with M. shawi in ZCL endemic areas. 24 1 Ege University Institute of Health Sciences, Bornova, İzmir, Turkey; 2Ege University Faculty of Science, Department of Zoology, Bornova, İzmir, Turkey; 3Akdeniz University Faculty of Science, Department of Biology, Antalya, Turkey; 4Celal Bayar University Vocational School of Health Sciences, Manisa, Turkey; 5Ministry of Health Public Health Agency, Antalya, Turkey; 6Ege University Faculty of Medicine, Department of Parasitology, Bornova, İzmir, Turkey; 7Celal Bayar University Faculty of Medicine, Department of Parasitology, Manisa, Turkey; 8Ege University Faculty of L i t e r a t u re , D e p a r t m e n t o f G e o g r a p h y, B o r n o v a , İ z m i r, Tu r k e y. [email protected] Keywords: Insecticide, Bednet, Antalya, Turkey Sand fly-borne diseases like visceral, cutaneous leishmaniasis and phlebovirus infections have been seen endemically in Turkey. The application of insecticides is mainly targeting mosquitoes/flies control. However, no study was performed about the susceptibility of sand flies against pyrethroids in Turkey. This study was carried out in Antalya province located in The Mediterranean Region of Turkey and we aimed to determine (1) the sand fly fauna, (2) the susceptibility of wild caught sand flies, mainly collected from rural areas, against commonly used pyretroids and (3) natural infection of Leishmania. CDC's bottle bioassay has applied for pyrethroid susceptibility by using different dosages. The tests were repeated 5 times with 100 specimens per bottle. In addition, WHO's Cone test were performed using two different bed nets (Olyset Plus® with PBO and Permanet®) to understand their efficacy. Following the standards of WHO testing procedures, all specimens were dissected, mounted and identified using Mediterranean keys. After dissection, the rest of the body of the specimens was pooled according to the locations and species and totally 50 pools were generated. DNA extractions were made using commercial kit and previously described conventional PCR of ITS-1 region was performed. A total of 10 Phlebotomus species were found in the province with the abundance of P. neglectus (38.82%), P. alexandri (21.66%) and P. tobbi (20.44%). Totally three different active ingredients were tested and knock down times were calculated using Probit Analysis. Knock down times were noted with 10min intervals, KdT50/KdT100 were noted for determining the effective dosages. For the lowest dosage of insecticides, KdT100 value were noted as 36 minutes for permethrin (0.05%), 60 minutes for cypermethrin (0.05%) and 42 minutes for deltamethrin (0.05%). By the end of 24-hour period, no sand fly was alive in bottle assay while 46% and 0% of sand flies were alive in cone test-Permanet® and cone test- Olyset Plus®. Four pools containing Phlebotomus neglectus and one pool Phlebotomus tobbi were found to be positive for Leishmania spp. In conclusion, no resistance was determined in bottle assay but upcoming resistance with prolonging death time and knock down times were noted. Cone test results have showed that Olyset Plus® has more knockdown effect than Permanet®. Mediterranean Region is one of the important tourism and agriculture centers in Turkey. Related to this the insecticide usage. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 5-O Vector Control ID 6-O Vector Control SAND FLY CONTROL IN BIHAR, INDIA: MODELING ALTERNATIVE CONTROL STRATEGIES USING SYSTEMIC INSECTICIDES ORALLY ADMINISTERED TO LIVESTOCK DEVELOPMENT OF RESISTANCE AGAINST DDT TO PHLEBOTOMUS ARGENTIPES (DIPTERA: PSYCHODIDAE) AND SEARCH FOR ALTERNATIVE INSECTICIDE David M Poché, William E Grant, Hsiao-Hsuan Wang Diwakar Singh Dinesh, Faizan Hassan, Vijay Kumar and Pradeep Das Texas A&M University, Genesis Laboratories, Inc. Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna-800007, India. [email protected] 25 [email protected] Keywords: Bihar, fipronil, Phlebotomus argentipes, simulation model. Approximately 60 percent of visceral leishmaniasis (VL) cases occur in Northern India and bordering regions of Nepal and Bangladesh, where the disease is spread by the sand fly species Phlebotomus argentipes. Bihar is the most poverty-stricken state in India and the majority of VL cases reported in the country occur in this region. DNA extraction suggests that cattle, domestic buffalo, goats, and humans serve as the primary hosts for P. argentipes in villages in Bihar. Recent research in Bihar has indicated that the systemic insecticide fipronil, orally administered to cattle, can kill 100% of adult P. argentipes feeding on insecticide-treated cattle and also kill 100% of larvae feeding on feces from insecticide-treated cattle. In addition, fipronil is a lipophilic compound and has the unique ability to be efficacious for several weeks. These results have suggested that fipronil can serve as a means for controlling sand fly populations and potentially reduce cases of VL in the region. To evaluate the potential efficacy of large scale fipronil treatment, a quantitative model was developed, representing the various sand fly life stages (egg, larva, pupa, and adult) in a typical rural village in Bihar. Seasonal variations in temperature, precipitation and relative humidity were incorporated into the model to better represent P. argentipes seasonality in Bihar within the system of interest. A variety of control strategies were simulated in which we varied the season and frequency of livestock treatment. Simulation results suggest that the potential to reduce the sand fly population in Bihar will depend on coordination of the timing of treatments with particular phases of the P. argentipes life cycle as they relate to sand fly seasonality in Bihar. The simulations also suggest that control strategies should be administered over a multiyear period to achieve preferred results. Keywords: Sand fly DDT resistant Phlebotomus argentipes (Diptera: Psychodidae) Annandale and Brunetti, the vector of Visceral Leishmaniasis in Bihar, India is developing resistance against the insecticide of choice, i.e. DDT. The measurement of tolerance/resistance was based on tube-bioassay test with 4% DDT impregnated paper to the laboratory bred newly emerged unfed and blood fed along with wild caught fed female P. argentipes. Variable results were found showing 40-60% resistant population in all tested P. argentipes. This was verified and compared with changes in Glutathion S- transferase (GST) enzyme (major detoxifying enzyme for xenobiotics) at protein level. The diagnostic band of 42 kD was found highly expressed among the resistant group by the 12% SDS- PAGE in the lab colonized and wild caught P. argentipes that was also confirmed as GST through Western Blot. Other enzymes like Acetylcholinesterase, Esterase and Mono oxygenase enzymes are under process of evaluation. Further, alternative insecticide like Deltamethrin was tried and found very effective in killing P. argentipes in tube bioassay test conducted with 0.05% WHO impregnated paper corresponding with the result of indoor residual spray 20mg/m2. This study will lead in finding actual number of resistant population along with the use of an alternative effective insecticide to control transmission of the disease. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 7-O Vector Control ID 8-O Vector Control T O X I C A N D B E H AV I O U R A L E F F E C T S I N EXPERIMENTAL HUTS OF LONG LASTING INSECTICIDE TREATED NETS AGAINST LUTZOMYIA LONGIFLOCOSA IN THE SUB-ANDEAN RURAL ZONE OF COLOMBIA A CONTROL STRATEGY FOR OLD WORLD CUTANEOUS LEISHMANIASIS OUTBREAKS Erika Santamaría, Olga Lucía Cabrera, Raul H Pardo Grupo de Entomología, Instituto Nacional de Salud, Bogotá. Grupo de Entomología y Enfermedades Transmitidas por Vectores, Universidad de La Salle, Bogotá. [email protected] 26 Waleed Al Salem1,2*, Louise Kelly-Hope1,3, Maha Abdeladhim4, Salah Balgonaeem2, Mohammed Al Zahrani2, Jesús Valenzuela4, Álvaro Acosta-Serrano1,5 1 Parasitology Department, LSTM, England, 2Saudi Ministry of Health, Riyadh, Saudi Arabia, 3Neglected Tropical Disease Centre, LSTM, England, 4Vector Molecular Biology Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Maryland, USA, 5Vector Biology Department, LSTM, England. [email protected] Keywords: Lutzomyia longiflocosa, long lasting insecticide treated nets, efficacy, experimental hut Keywords: Old world cutaneous leishmaniasis, Saudi Arabia, Al Ahsa The aim of the study was to evaluate the toxic and behavioral effects of long-lasting insecticide-treated nets (LLNs) against Lutzomyia longiflocosa in the sub-Andean rural zone of Colombia in the first experimental hut designed for sand flies. Three bednets were tested: Permanet 2.0 (a.i.: deltamethrin, 55 mg/m2), bednet treated with Icon Maxx (a.i.: lambda-cyhalothrin, 60 mg/m2), and control (untreated bednet). Several holes were made on the bednet to simulate the conditions of a torn net. In each hut, two volunteers were located inside the bednet. The study design was a 3 x 3 multiple Latin square. The recorded variables were: number of sand flies inside the hut and within the exit traps, human landing rate, and mortality. The detected effects of the LLNs on L. longiflocosa were: 1) Inhibition from entering the net: the percentages of females found inside the nets treated with Icon Maxx (13.9%) and Permanet (2.0, 44.8%) were significantly lower compared to the control (77.1%). The Icon Maxx showed the greatest difference in percentage of females inside the net compared with the control, 62.3% (59.0 - 69.4), twice the difference in percentage of females found inside the Permanet, 32.3% (27.7- 36.9). 2) Landing inhibition: The mean human landing rate was 10 times [4.9 females/night/2 persons (f/n/2 p)] and three times (15.1 f/n/2 p) lower in the Icon Maxx and Permanet (2.0) respectively, compared with the control, 49.5 f/n/2 p. The protective effect of the Icon Maxx (90.1%) was higher compared to the Permanet (69.5%. 3) Repellency: the mean exit rate of L. longiflocosa was significantly higher in the Icon Maxx (14.1 f/n) compared to the control (5.1 f/n). Furthermore, the percentage of resting females on the walls of the hut with the LLNs (using as denominator the total number of females outside the net) was significantly lower with the Icon Maxx (23.3%) and Permanet (30.5%) compared to the control (72.3%. 4). Mortality: Mortality at 0 h post-test with the Icon Maxx was almost twice (52.5%) that recorded with the Permanet (28.6%). Nevertheless, mortality at 24 h was similar for both LLNs (78.8% and 82.6%) respectively, without significant differences. The two LLNs showed strong effects on L. longiflocosa behavior and mortality. However, the Icon Maxx seems better because of its quicker lethal effect, higher protective effect, inhibition from entering the net and repellency. Old World cutaneous leishmaniasis (CL) is one of the most prevalent vector-borne diseases in the East Mediterranean Region, including the Kingdom of Saudi Arabia (KSA). A combination of factors is responsible for the sustained spread of CL throughout this region, including lack of health education, socio-economic factors, civil unrest, and human migration. Furthermore, uncontrolled urbanization and lack of governmental sector integration (i.e. Ministries of Health and Agriculture, and Municipalities) impede the establishment of a coordinated disease control strategy. Our case study focused on a new construction site in a remote area of Al Ahsa governorate (Eastern KSA), which had not been previously assessed for CL by the local leishmaniasis control team. This region is known for being exclusively endemic for zoonotic CL (1, 2). Prior to the start of the leishmaniasis transmission season (~April 2012), 150 (nonlocal) construction laborers, with no previous history of CL, arrived at the construction site. In January 2013, ~60% of the construction laborers were reported to have acquired CL, and further PCR-RFLP analysis confirmed they were infected with Leishmania major (2). Furthermore, the same cohort of patients presented higher serum levels of anti-SP32 antibodies (a known Phlebotomus papatasi saliva marker (3)) compared to infected local residents living in neighboring CL-controlled areas. Over the course of 2013 (March-November), an integrated disease control strategy, conducted by several governmental sectors, which consisted of rodent (mechanical) and vector control, was applied in and around the construction site. Mechanical control involved the removal of Hammada elegans (rodent food plant) and destruction of rodent burrows within a radius of 500 m around the construction area. In addition, vector control was employed by spraying deltamethrin into rodent burrows and wall cracks. Since the implementation of this integrated control strategy, no CL cases have been reported at the construction area up until April 2014.This case study suggests that (i) implementation of an integrated vector and rodent control program is sufficient to overcome outbreaks of zoonotic Old World CL and, (ii) assessing potential disease transmission areas prior to the establishment of new development, is essential to prevent CL outbreaks. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 27 ID 9-O Vector Control ID 10-O Vector Control LONG LASTING INSECTICIDE TREATED NETS DECREASE SANDFLY ABUNDANCE AND BLOOD FEEDING SUCCESS INDOORS IN THE SUB-ANDEAN REGION OF COLOMBIA: RESULTS OF A HAMLET VECTOR CONTROL TRIAL AFTER ONE YEAR FOLLOW UP FIRST EXPERIMENTAL HUT TO STUDY THE EFFECT OF INSECTICIDE CONTROL MEASURES AGAINST SANDFLIES (DIPTERA: PSYCHODIDAE) Olga Lucía Cabrera, Erika Santamaría, Raúl H Pardo Instituto Nacional de Salud Raúl H Pardo, Olga Cabrera, Catalina Marceló, Erika Santamaría [email protected] Universidad de La Salle. [email protected] Keywords: experimental hut, sandflies, control Keywords: control, Lutzomyia longiflocosa, bednets Experimental huts have been used to evaluate the effects of insecticide control measures against mosquitoes indoors mainly for malaria vectors. Such huts have not been used for sand flies. The aim of this study was to build an experimental hut in which to evaluate insecticide control measures against sand flies. The study was carried out in the sub-Andean coffee area of Colombia in an endemic area for cutaneous leishmaniasis (CL) where Lutzomyia longiflocosa is abundant. The project was conducted in three parts: 1) Avenues of entrance and exit. To identify the main avenues of entrance and exit into a rural house, two observational studies on the entering and exiting behavior of sand flies were conducted. The resulting data were used to determine trap placement in the experimental hut. For entrance sites the numbers of sand flies attempting to enter the house through large openings (sticky trap catches) and narrow slits (cage trap catches) were compared; while for exit sites the numbers of blood-fed females previously released within the house, trying to escape through the openings described above were compared. The results from both experiments indicate that L. longiflocosa enter (95% of 313 females) and exit (96% of 177 females) the house mainly by large openings between the ceiling and walls; 2) Trap selection. To select the type of traps to use in the hut, laboratory tests were conducted with Lutzomyia longipalpis in a wind tunnel. Two traps, modified for collecting sand flies were evaluated based on their ability to allow the sand flies to enter and prevent them from exiting the trap. The results show that the horizontal slit trap was the best trap, allowing 84.1% of females to enter and preventing 87.3% of those that entered from escaping; 3) Hut construction. Based on the afore mentioned results, three experimental huts were built in the field. Each hut was a 2.5x2.5x2.3 m room made of bricks with metal roof, ceiling and two large openings in each wall, between the ceiling and the wall, at which to locate the horizontal slit trap system. Special care was taken to ensure to block all other possible escape sites; and 4) Experimental hut test. Hermeticism was evaluated by releasing 300 females inside each hut. After eight hours 70% of the released females were recaptured. Observations of human landing catches of L. longiflocosa inside the experimental huts compared with a control house (hut without traps) showed that L. longiflocosa entered the experimental huts in lower numbers than the control house. By adding baffle traps to the entrances of the experimental huts it was shown that sand flies entered the experimental hut and the control in similar densities. The aim of the study was to evaluate the entomological effect of long lasting insecticide treated bednet (LLNs) against Lutzomyia longiflocosa, a presumed main vector of cutaneous leishmaniasis in Colombia. The study was carried out in the mountainous coffee area of Huila department. The evaluated variables were: sandfly female abundance, blood feeding rate, feeding success (more than 75% of abdomen filled with blood) and mortality. The study was carried out at the hamlet level using a randomized block design, were 7 triplets of hamlets were formed based on pre-intervention sandfly abundance. Within each triplet three treatments were assigned: (i) Permanet (industrially treated), (ii) Icon Maxx (home treated) and (iii) control (untreated bednet). The study had three phases: 1) baseline study. 21 selected hamlets were visited to collect pre-intervention data, including sandfly abundance. Sandflies were collected by CDC light traps in at least 50% of houses and forest, 2) Intervention. Around 10% of the highest sandflies abundance houses were selected as representative for each hamlet, all beds in each hamlet were covered with a bednet, and 3) pos-intervention. Sandflies abundance was monitored at 1, 6 and 12 month time-points pos-intervention. Preintervention L. longiflocosa indoors mean abundance was similar by treatment. Total abundance pos-intervention decrease significantly from 14.7 f/trap/night in the control to 5.0 f/trap/night for Icon Maxx and 4.3 f/trap/night for Permanet. It means a reduction of around 70% in sandfly abundance in each of the LLNs. Comparison for each of the three month time-points showed the same significant reduction. Percentages of blood feeding success for the pre-intervention were significantly different between treatments, so comparisons preintervention vs. pos-intervention were made for each treatment. Percentage of blood feeding success decreased significantly in the LLNs with a reduction of 41.5% for Icon Maxx and 28.1% for Permanet, while the control had a significant increase of 84%. Sandflies abundance in forest was apparently reduced in the treatment with Permanet but it increased in the treatment with Icon maxx. The two LLNs reduce sandfly abundance indoors and blood feeding success in L. longiflocosa. Icon Maxx could be a useful alternative to become untreated bednets and treated bednets with reduced insecticide power to LLNs. Mass killing effect of LLNs against sandflies could not be discarded. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 11-O Vector Control ID 12-O Vector Control EFFICACY OF IMPREGNATED BEDNETS AND SPECIES COMPOSITION IN PREVENTION OF CANINE LEISHMANIOSIS IN NATURALLY EXPOSED DOGS USING A 10% IMIDACLOPRID/ 4.5% FLUMETHRIN COLLAR (SERESTO®) EXPERIMENTAL HENHOUSES Manteca Acosta Ma, b; Molina Ja; Utgés M Ea, b; Mastrangelo AVb, c; Pérez AAd; Santini M S.b; Salomón ODa, c REDILA a Instituto Nacional de Medicina Tropical (INMeT); b Centro Nacional de Diagnóstico e Investigación en Endemo-epidemias (CeNDIE); c Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); d Departamento de Ecología, Genética y Evolución. FCEyN (UBA). 28 Domenico Otranto1; Filipe Dantas-Torres1,2; Emanuele Brianti3; Fabrizio Solari Basano4; Dorothee Stanneck5; Katrin Deuster5 1 University of Bari, Italy; 2Oswaldo Cruz Foundation, Brazil; 3University of Messina, Italy; 4Acroblu s.r.l., Italy; 5Bayer Animal Health GmbH, Germany. [email protected] [email protected] Keywords: impregnated fabrics, vectorial control, sandfly The use of insecticide-impregnated fabrics as preventive tools has become a relevant topic of research in the epidemiology and control of vector-borne diseases because of their ability to reduce vector-human contact and acceptability by end users at the local level in communitybased vector management programs. In this study we used noncommercial curtains with the intent of finding a participative control strategy where users can make and use the curtains in their own community. The project was developed in a rural area (Misiones, northeastern Argentina) where epidemic outbreaks of cutaneous leishmaniasis (CL) occurred in 2004-2005 vectored by Nyssomyia whitmani. The objective was to evaluate changes in sand fly abundance, particularly of Ny. whitmani, in the presence of 10% permethrine-impregnated mosquito-nets under semi-field conditions. Three experimental henhouses were installed on the border of the forest, each with three chickens. The treatments (impregnated curtain, IC) and controls (non-impregnated curtain, NIC; without curtain, WC) were assigned randomly. To simulate incomplete coverage of human dwellings, curtains were hung so as to cover only two sides of the henhouses. During two consecutive nights, a light trap was set inside each henhouse and in a site without chickens (WCH) from 5 pm to 9 am to measure sand fly abundance.). Trials were repeated monthly from February 2012 to January 2013 except for the period April-June when sand fly populations were extremely low. All sand flies were counted to calculate total abundance and a 10% sample of each combination of month and treatment was used to determine species relative abundance. A total of 38,277 sand flies was collected representing eight species )as follows: Ny. whitmani (89.9%), Migonemyia migonei (5%), Pintomyia pessoai (2.7%), Bruptomyia sp. (1.7%), Lutzomyia longipalpis (0.4%); Evandromyia corteleziisallesi, Psathyromyia shannoni and Lutzomyia monticola (<0.1%). Nyssomyia whitmani was the most abundant species regardless of treatment (>85%). The trap index (average number of sand flies collected/trap/night) per treatment was IC (63.6, SEM 21.5) and NIC (160.2, SEM 62.3), both lower than WC (454.2, SEM 160.1) (F3,14=5.6, p=0.005). We did not find differences between IC and NIC, nor between IC and WCH (59.3, SEM 19.2). These results show significant efficacy both in the use of the impregnated and nonimpregnated curtains as preventive barriers to reduce human-sand fly contact. Keywords: Prevention, canine leishmaniosis, imidacloprid, flumethrin Field evaluations were conducted to investigate the protective efficacy of a collar containing 10% imidacloprid/ 4.5% flumethrin (Seresto®) for the prevention of Leishmania infantum infection in dogs at different time points (2011-2013). Three animal shelters, two in Sicily and one in Apulia, located in hyper-endemic areas for L. infantum with confirmed presence of competent vectors were selected and 340 dogs testing negative by serology (IFAT) and PCR on skin and bone marrow were enrolled. Dogs were randomly allocated to collar-treatment (165) or untreated control (175) group. The collars were applied at the beginning of the transmission seasons according to label instructions and only replaced if lost or required for body weight changes. Blood, bone marrow and skin samples were collected throughout the study and at the final follow-up. Serological, cytological and molecular tests were performed to detect the presence of L. infantum in the different tissues. In Apulia, dogs were ≤6 months of age at enrolment and of various breeds and had not been exposed to sand flies previously. None of the collar-treated dogs tested positive for L. infantum throughout follow-up, whereas 21 of 58 untreated control dogs tested positive throughout the follow-up with five animals also displaying clinical signs at the final time point. The cumulative incidence density rate (IDR) observed in untreated animals was 46.2%. In Sicily, dogs were 2 months to 8 years old at enrollment and of various breeds. Three of 102 dogs in the collartreatment group and 47 of 117 untreated control dogs tested positive throughout the follow-up. The cumulative IDR observed in untreated animals was 60.7%. The collar (10% imidacloprid/ 4.5% flumethrin) was demonstrated to be safe and highly effective (93.4–100%, based on IDRs) in preventing L. infantum infection throughout the season in heterogeneous dog populations under varying environmental conditions. The regular use of treated collars during the sand fly season thus represents a reliable and sustainable strategy for the control of leishmaniasis in dogs living in or travelling to endemic areas. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 13-O Taxonomy & Phylogeny ID 14-O Taxonomy & Phylogeny A NEW PROPOSAL FOR THE TAXONOMIC IDENTIFICATION OF PHLEBOTOMINE SAND FLIES (DIPTERA: PHLEBOTOMINAE) IN BRAZIL SEX PHEROMONES AS A PRE-MATING SPECIES ISOLATION BARRIER IN BRAZIL 1, 3 2 1 Andrey J de Andrade , Paloma H F Shimabukuro , Eunice A B Galati 1 Departamento de Epidemiologia, Faculdade de Saúde Pública, Universidade de São Paulo, Av. Dr. Arnaldo, 515, Pinheiros, São Paulo, SP, 01246-904, Brazil, 2Centro de Referência Nacional e Internacional para Flebotomíneos, Centro de Pesquisas René Rachou, Instituto Oswaldo Cruz, Av. Augusto de Lima, 1715, Barro Preto, Belo Horizonte, MG, 30190-002, Brazil, 3Laboratório de Parasitologia Médica e Biologia de Vetores, Área de Patologia, Faculdade de Medicina, Universidade de Brasília, Campus Universitário Darcy Ribeiro, Asa Norte, 70910-900, Distrito Federal, Brazil. 29 V Carter1, DP Bray1, G Borges-Alves1, E Dilger2, O Courtenay2, R Brazil3 and JGC Hamilton1 1 Centre for Applied Entomology and Parasitology, School of Life Sciences, Huxley Building, Keele University, Staffordshire, ST5 5BG, United Kingdom; 2School of Life Sciences, University of Warwick, Coventry, CV4 7AL, United Kingdom; 3Fundaçao Oswaldo Cruz, Instituto Oswaldo Cruz, Lab Doenças Parasitarias, Rio de Janeiro, Brazil. [email protected] [email protected] Keywords: pheromone, species complex, reproductive barrier Keywords: identification key, taxonomy, type-species, Brazilian sand fly species Currently, there are 269 valid phlebotomine sand flies species recorded in Brazil, which represents the highest diversity of sand flies when compared to other South American countries. Most species (approximately 110) were described between 1930 and 1950 by Brazilian researchers following different classifications and taxonomic proposals. Systematics of sand flies has been debated during the last decades, new classifications have been proposed, and relationships between these species are being studied based on morphological and molecular characters. Dichotomous identification keys are the first step towards specific identification and for Brazilian sand flies the last identification key was published in 2003. The aim of this study is to present an updated new key to indentify sand fly species recorded in Brazil. To produce the key type-species deposited in nine Brazilian institutions were examined. One hundred and sixty nine species including males and females were found. Observations were made regarding the preservation status of the slides - whether the specimens were still visible and labels were adequately presented; the presence of microscopic slides according to the original description; and specimens numbers and typology. All type-species were photographed to build an image dataset. When types were not found, additional specimens of particular collections were examined. Drawings of the genitalia were made using a camera lucida and covered by ink pen. A list of geographical distribution for Brazilian states, including taxonomic synonyms, and a taxonomic catalog were compiled. The key is divided by gender and the species are grouped by morphological characters rather than by phylogenetic relationships. A total of 251 and 179 dichotomies for 258 males and 235 females, respectively, are presented in the key. Researchers of public institutions and health services will be invited to test the taxonomic key, so it can be validated. An interactive version of this key will be produced using the DELTA (Description Language for Taxonomy) software package. It will include all data compiled by us. The DELTA system allows a quick update as new species will be recorded and described in Brazil. Financial support: FAPESP. Members of the Lutzomyia longipalpis species complex in Brazil can be distinguished through morphological, physiological and biochemical markers. Determining the relative importance of the different members of the L. longipalpis species complex in transmitting visceral leishmaniasis is important for understanding the epidemiology and control of the disease. It has been proposed that the species complex is under an incipient speciation process, and one factor influencing speciation is mate-finding through male sex pheromones. Male L. longipalpis may be one of four possible sex pheromone chemotypes: (S)-9-methylgermacrene-B (9MGB), 3methyl-alpha-himachalene, cembrene 1 or cembrene 2. Previous studies of crosses between different chemotypes carried out in the laboratory showed differing degrees of reproductive isolation. Thus the sex pheromones of L. longipalpis could affect pre-mating behaviour and prevent random mating and gene flow between individuals. However, field evidence for pheromones acting as premating barriers is lacking. We investigated the cross-attraction of L. longipalpis, from two different pheromone chemotypes in Brazil (9MGB from Araçatuba, Sao Paulo State and cembrene 1 from Marajo, Para State). Using experimental chicken boxes and pheromone lures containing either pheromone or blank controls, CDC light traps were used to capture sand flies attracted to the site. Both male and female sand flies were significantly more attracted to their conspecific pheromone chemotype than CDC traps containing no pheromone. Sand flies were not significantly attracted to sex pheromone of the alternative chemotype. These results support the contention that male sand fly sex pheromones act as a pre-mating isolation barrier in the field, reduce non-productive mating encounters and therefore may be one of the strongest influences on speciation in the L. longipalpis complex. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 15-O Taxonomy & Phylogeny ID 16-O Taxonomy & Phylogeny IDENTIFICATION OF PERUVIAN SAND FLY SPECIES (DIPTERA: PSYCHODIDAE) THROUGH DNA BARCODES ORIGINS AND AFFINITIES OF MALAGASY PHLEBOTOMINE SANDFLIES 1 2,3 Chukwunonso Onyemaechi Nzelu , Abraham G Cáceres , Ken Katakura1, Yoshihisa Hashiguchi4,5,6 and Hirotomo Kato1 1 Laboratory of Parasitology, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan; 2Departamento Académico de Microbiología Médica, Facultad de Medicina Humana, Universidad Nacional Mayor de San Marcos, Peru; 3Laboratorio de Entomología, Instituto Nacional de Salud, Peru; 4Department of Parasitology, Kochi Medical School, Kochi University, Japan; 5Prometeo, Secretaria Nacional de Educacion Superior, Ciencia, Tecnologia e Innovacion (SENESCYT), Ecuador; 6Centro de Biomedicina, Universidad Central del Ecuador, Ecuador. 30 Randrianambinintsoa Fano José1; Léger Nicole2; Boyer Sébastien1; Robert Vincent3; Depaquit. Jérôme2 1 Unité d'Entomologie Médicale, Institut Pasteur de Madagascar, Antananarivo, Madagascar. 2Université de Reims Champagne-Ardenne, ANSES, EA4688-USC « VECPAR », Reims, France. 3MIVEGEC, UMR, Montpellier, France. [email protected] Keywords: Phlebotominae, Biogeography, Madagascar, Systematics [email protected] Keywords: molecular taxonomy, DNA barcode, sand flies, vectors. Understanding the ecological and medical importance of sand flies relies on accurate identification of species. Currently, the identification of phlebotomine sand flies is based on morphological characteristics. However, this traditional taxonomy is laborious, time consuming and often complicated by cryptic species complexes. Molecular approaches have become increasingly employed and promise to be useful taxonomic markers for rapid and reliable species identification. The use of DNA barcodes has been proposed recently as a tool for identification of the species in different animal taxa. In this study, we explored the efficiency of DNA barcode approach, based on cytochrome c oxidase subunit I (COI) sequences, for species identification of Peruvian sand flies, which included major vector species. A total of 89 sand fly specimens belonging to sixteen morphological species and two genera (Lutzomyia and Warileya) were analyzed. We were able to recover and align the target COI fragment from all sand fly species examined. Neighbor-joining (NJ) analysis of the DNA barcode sequences show that each sand fly species formed barcode clusters with tight cohesion that were clearly distinct from those of allied species and were largely congruent with the conventional taxonomy. Thus, the results validated the usefulness of DNA barcoding systems in species recognition in sand flies from Peru. Until now, the Phlebotomine sand fly fauna (Diptera, Psychodidae, Phlebotominae) of Madagascar remained largely unexplored and unknown according to the lack of autochthonous transmission of leishmaniasis in this country. To date, three phlebotomine sandflies genera including three endemic subgenera have been recorded from Madagascar Island.: fourteen species are already described. Since 2000, field trap phlebotomine sandflies were studied using both morphology and molecular biology (rDNA and mtDNA).One objective is to understand the origins and the affinities of this fauna group that was never studied due to the lack of precise knowledge on the origins of different lineages of these sandflies. At the light of the results obtained including molecular characterization and phylogenies, the authors discuss the endemism concerning Phlebotomine sandflies from Madagascar, the relationships of Malagasy phlebotomine sand flies with species from Africa, America, Oceania and Asia, and propose an hypotheses explaining the settlement of this island by Phlebotomine sandflies, including an old one related to the isolation of Madagascar from the Gondwana following a generalized track compatible for the Malagasy Phlebotomus and probably the Vattieromyia. The morphological affinities of the later subgenus with two Australian species S. pugifera et S. standfasti or with the American Pressatia is discussed. The individualization of the Trouilletomyia and Grassomyia should be younger and underlined several routes of settlement of Madagascar. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 31 ID 17-O Taxonomy & Phylogeny ID 18-O Taxonomy & Phylogeny DNA BARCODING OF MEDITERRANEAN PHLEBOTOMINE SANDFLIES BASED ON THE SEQUENCE ANALYSIS OF THE MITOCHONDRIAL CYTOCHROME OXIDASE I GENE A MOLECULAR PHYLOGENY OF THE PHLEBOTOMINAE Jérôme Depaquit, Maria Dolores Bargues, Santiago Mas-Coma Israel Cruz, Rosa Gálvez, Oihane Martín, Ivonne Pamela LlanesAcevedo, Carolina Arcones, Rocío Checa, Ana Montoya, Carmen Chicharro, Silvia Miguelañez, Guadalupe Miró WHO Collaborating Center for Leishmaniasis, Servicio de Parasitología, Centro Nacional de Microbiología, Instituto de Salud Carlos III. Ctra. Majadahonda-Pozuelo Km2, 28220 Majadahonda-Madrid, SPAIN Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad Complutense de Madrid. Avda. Puerta de Hierro s/n, 28040 Madrid, SPAIN. [email protected] Keywords: Sandflies, Mediterranean, DNA-barcoding, COI Genomic approaches to taxonomy rely on the variability of DNA sequences to identify organisms. In this sense, these sequences can be considered as genetic barcodes. Mitochondrial genes seem to be the most appropriate candidates for DNA barcoding, among them cytochrome c oxidase I (COI) possess a greater range of phylogenetic signal than any other mitochondrial gene and it has been suggested as the core of a global bioidentification system for animals. DNA barcoding is increasingly used in phlebotomine sandflies taxonomy, using either COI or cytochrome-B (cytb) genes. We aimed to assess the use of COI-based DNA barcoding using sandflies collected from different regions in Spain, for which we sequenced a 658 bp fragment of the COI gene. We completed this data set with DNA sequences available in the GenBank for species prevalent in the Mediterranean region. In total 156 DNA sequences from 4 different phlebotomine sandflies species (Phlebotomus ariasi, P.papatasi, P. perniciosus, and Sergentomyia minuta) from Spain were generated for this work and added to 129 DNA sequences from 14 different phlebotomine species (P. alexandri, P. ariasi, P. bergeroti, P. chadlii, P. longicuspis, P. papatasi, P. perfiliewi, P. perniciosus, P. riouxi, P. sergenti, S. antenata, S. clydei, S. fallax, S. minuta), from 6 different countries (Algeria, India, Italy, Malta, Portugal, Tunisia) obtained from the GenBank. Sequences were aligned and trimmed to get a final alignment of 620 nt. Intraspecific variability was observed in all species but P. bergeroti (a single DNA sequence was available), and haplotypes were defined on the basis of single nucleotide polymorphisms. Distances were estimated with the Jukes Cantor model to confirm the data set was amenable for Neighbor-Joining analysis. With the refined alignment a Neighbor-Joining Tree was obtained using Kimura 2-parameters model of evolution and 2000 bootstrap replicates. All species included in the tree were supported by bootstrap values of 98% or higher, indicating that this approach is a solid candidate for DNA barcoding of phlebotomine sandflies. This work represents a strong contribution to the collection of COI DNA sequences available for phlebotomine sandflies from Spain. We also discuss different approaches to DNA barcoding using other targets, such as cytb, as well as other analysis methods and models of evolution. Acknowledgement: COLCIENCIAS/COLFUTURO 2012, Instituto de Salud Carlos III MPY-1248/12. Université de Reims Champagne-Ardenne, ANSES, EA4688 – USC. [email protected] Keywords: Phylogeny, ribosomal DNA, paleobiogeography. Phlebotomine sandflies belong to the subfamily Phlebotominae (Diptera, Psychodidae). The systematics of this group is the inheritance of many typological systematics carried out in the Old World (Abonnenc 1972, Lewis 1982, Artemiev and Neronov 1984, Seccombe, Ready et al. 1993) and in the New World (Young and Duncan 1994 ). However, a few phylogenetical and phenetical studies at the subfamily scale have been carried out (Galati 1995, Rispail and Léger 1998, Rispail and Léger 1998, Galati 2010) based on morphological characters.We propose a molecular phylogeny of the Phlebotominae based on specimens from Africa, Europe, Asia, Oceania and the Americas. Our sampling includes the following genera: Warileya, Spelaeophlebotomus, Idiophlebotomus and Chinius regarding the Hertigiini; Phlebotomus (subgenera Phlebotomus, Paraphlebotomus, Synphlebotomus, Larroussius, Adlerius, Transphlebotomus, Euphlebotomus, Anaphlebotomus, M a d a p h l e b o t o m u s , L e g e ro m y i a , a n d A b o n n e n c i u s ) , Australophlebotomus, Brumptomyia, Oligodontomyia, Sergentomyia (Sergentomyia, Parrotomyia, Sintonius, Rondanomyia, Vattieromyia, Trouilletomyia and some ungrouped species), Grassomyia, Spelaeomyia, Parvidens, Daenemyia, Edentomyia, Micropygomyia (Silvamyia, Sauromyia, Micropygomyia), Sciopemyia, Lutzomyia (Helcocyrtomyia, Castromyia, Tricholateralis, Lutzomyia), Migonemyia (Migonemyia, Blancasmyia), Pintomyia (Pintomyia, Pifanomyia), Dampfomyia (Coromyia), Expapillata, Pressatia, Trichopygomyia, Evandromyia (Aldamyia, Evandromyia, Barrettomyia), Psathyromyia (Foratiniella, Xiphomyia, Psathyromyia), Viannamyia, Martinsmyia, Bichromomyia, Psychodopygus, Nyssomyia and Trichophoromyia regarding the Phlebotominii. We have chosen the following outgroups: Psychoda sp. Sycorax sp., Nemapalpus spp and Bruchomyia sp.We sequenced the complete small subunit of the ribosomal DNA and also several markers of the long subunit (D1, D2 and D8 domains).We compared our results with those previously published based on morphological characters (Galati 1995, Rispail and Léger 1998, Rispail and Léger 1998, Galati 2010). Our data will be of help to discuss the systematics of the Phlebotomine sandflies. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 19-O Taxonomy & Phylogeny ID 20-O Taxonomy & Phylogeny DISTRIBUTION, PHYLOGENY AND TAXONOMY OF SERGENTOMYIA IN ETHIOPIA RAPID IDENTIFICATION OF PHLEBOTOMINE SAND FLIES BY MALDI-TOF MASS SPECTROMETRY Aysheshm Kassahun1, Kristýna Hlavačková1, Vít Dvořák1, Asrat Hailu2, Alon Warburg3, Jan Votýpka1 and Petr Volf1 Vit Dvořák1, Jérôme Depaquit2, Valentin Pflüger3, Petr Halada4, Kristýna Hlavačková1, Veronique Lehrter2, Jorian Prudhomme5, Sonja Zapata6, Petr Volf1, Alexander Mathis7 1 Charles University in Prague, Czech Republic; 2Addis Ababa University, Ethiopia; Hebrew University of Jerusalem, Israel. 3 [email protected] Keywords: Sergentomyia, leishmaniasis, S. schwetzi Members of the genus Sergentomyia are dominant sand flies in SubSaharan Africa. Several studies have demonstrated their importance as vectors of reptilian leishmaniasis caused by Sauroleishmania but their role, if any, in the epidemiology and transmission of human leishmaniasis is not well understood. Multiple cases of Sergentomyia feeding on mammals together with their abundance in the foci of human leishmaniasis and occasional records of Leishmania parasites within their bodies detected by microscopy or PCR have led to speculation on Sergentomyia involvement in the epidemiology of leishmaniasis. Recently, however, we experimentally demonstrated that S. schwetzi is refractory to Leishmania parasites infecting humans. To study the biology, species diversity, phylogeny, taxonomic status and distribution of Sergentomyia species in Ethiopia, sand flies have been collected since 2010 using CDC light traps and sticky traps in endemic areas, mainly in domestic and peri-domestic habitats (animal shelters, termite mounds, rodent burrows and caves and acacia and balanite trees). The specimens collected were identified morphologically and by molecular methods, particularly by the sequencing of cytochrome C oxidase I and cytochrome B. Seven Sergentomyia species were found. Males of two morphotypes of S. schwetzi differed in the orientation of the spines on the style, shape of the penis and size of the 6th abdominal tergite. Using laboratoryestablished colonies we investigated the effect of ambient temperature during larval development on the morphological traits of adults. The genetic variability and proteomic profile were assessed by sequencing target genes and analysing of the spectrum produced by MALDI-TOF (Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry), respectively. Moreover, we investigated the possibility of parthenogenesis and autogeny in S. schwetzi . This study was supported by the Bill and Melinda Gates foundation [grant number OPPGH5336] and Grant Agency of Charles University [9108/2014]. 32 1) Department of Parasitology, Faculty of Science, Charles University in Prague, Prague, Czech Republic. 2) Université de Reims Champagne-Ardenne, ANSES, EA4688 - USC "transmission vectorielle et épidémiosurveillance de maladies parasitaires (VECPAR)", Reims, France. 3) Mabritec SA, Riehen, Switzerland. 4) Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic. 5) MIVEGEC unit, Centre IRD, Montpellier, France. 6) Instituto de Microbiología, University San Francisco de Quito, Ecuador. 7) Swiss National Centre for Vector Entomology, Institute of Parasitology, University of Zurich, Zurich, Switzerland. [email protected] Keywords: MALDI-TOF, species identification, protein profiling Phlebotomine sand flies are vectors of several pathogens causing diseases in both humans and animals. To elucidate their role in transmission of these pathogens, proper and conclusive species identification is crucial. Since morphological determination is based on minute and sometime dubious characters on their head and genitalia, which may be altered or absent in field-collected material, there is a demand for rapid, simple and cost-effective complementary molecular approaches. Protein profiling using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDITOF MS) was established during the last decade as a routine method in clinical diagnostics of bacteria and fungi. Currently, applications of this method on higher organisms including arthropods are emerging. We demonstrated that MALDI-TOF MS is also feasible for species determination of phlebotomine sand flies. We established a reference database of protein spectra of 20 species from the Old World genera Phlebotomus (subgenera Phlebotomus, Paraphlebotomus, Larroussius, Adlerius and Euphlebotomus) and Sergentomyia as well as from four New World genera (Nyssomyia, Psychodopygus, Psathyromyia and Lutzomyia). Several major vectors of both cutaneous and visceral human leishmaniases are among analyzed species. Protein spectra originated from both sexes of laboratoryreared as well as field collected specimens. Best quality spectra were obtained with frozen specimens, but storage in 70% ethanol is feasible, and specimens stored for up to 25 years could be identified. In addition, we evaluated whether mass spectra obtained on the instruments of the two leading vendors (Bruker Daltonics, Germany; Shimadzu, Japan) can be used for species identification with the software of the other instrument. Such cross-applicability would strongly enhance the value of reference databases established at different institutions. The present study shows that protein profiling by MALDI-TOF MS is well applicable on phlebotomine sand flies and represents an analytical tool which offers high-throughput, sensitive and reliable species identification which is much desired for many applications in vector biology. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 21-O Taxonomy & Phylogeny ID 22-O Taxonomy & Phylogeny TWO NEW SPECIES OF THE SUBGENUS TRANSPHLEBOTOMUS REVEALED BY MOLECULAR TAXONOMY DNA SEQUENCE ANALYSIS SUGGESTS THAT CYTBND1 PCR-RFLP MAY NOT BE APPLICABLE TO S A N D F LY S P E C I E S I D E N T I F I C AT I O N THROUGHOUT THE MEDITERRANEAN REGION Vit Dvorak1, Ozge Erisoz Kasap2, Jerome Depaquit3, Bulent Alten2, Jan Votypka1, Petr Volf1 1 Department of Parasitology, Charles University in Prague, Prague, Czech Republic. 2 Department of Biology, Hacettepe University, Ankara, Turkey. 3Laboratoire de Parasitologie, Université de Reims, Ardenne, France. [email protected] Keywords: Transphlebotomus, molecular taxonomy, new species The subgenus Transphlebotomus comprises only three described species (P. mascittii, P. canaaniticus, P. economidesi) with markedly different geographical distribution. While P. mascittii is widely present in the north of the Mediterranean basin and recently recorded in Algeria, and stretches up to the northern limits of sand fly presence in Europe in localities in Belgium, Austria and Germany, P. canaaniticus and P. economidesi have more restricted distribution. Due to their similar morphology, proper identification remains difficult and relies mainly on molecular markers. This subgenus was long time neglected. The biology of its species is poorly understood, they are little represented in field collections as some of them are expected to be cavernicolous and are thought not to be incriminated in Leishmania parasites transmission, although closely related subgenera Larroussius and Adlerius harbor several proven vectors of both cutaneous and visceral leishmaniasis. However, presence of P. mascittii in several foci of autochtonous canine leishmaniasis in southern Germany suggests that Transphlebotomus species may be a vector under certain circumstances. We studied Transphlebotomus species from Crete, the largest island of the Aegean Archipelago and from a locality on south-western coast of Anatolia, approximately 500 km away from Crete. To clarify the taxonomic position of the specimens studied, P. mascittii from Belgium, Germany and France, P. economidesi from Cyprus and P. canaaniticus from Lebanon were used as reference sequences. Based on the sequencing analysis of several genes (cytochrome B, NADH dehydrogenase subunit 4, cytochrome oxidase I) two new putative species of this subgenus were found, both of them characterised also by following morphological characters. One species is described from males only and is mainly individualised by long third antennal segments coupled with a low number (13>-20) of coxal setae. The second one is described from both males and females. The latter are remarkable by their pharyngeal armature which is Adlerius-like more than Transphlebotomus-like. Moreover, P. economidesi was found in Turkey sympatrically with these two species, which is the first record of this species outside Cyprus. Our findings indicate that the taxonomy of the subgenus Transphlebotomus should be thoroughly studied by molecular and morphological methods together with investigations concerning possible vectorial capacity in Leishmania transmission. 33 Ivonne Pamela Llanes-Acevedo, Carolina Arcones, Rosa Gálvez, Oihane Martín, Rocío Checa, Ana Montoya, Carmen Chicharro, Silvia Miguelañez, Susana Cruz, Guadalupe Miró, Israel Cruz WHO Collaborating Center for Leishmaniasis, Servicio de Parasitología, Centro Nacional de Microbiología, Instituto de Salud Carlos III. Ctra. Majadahonda-Pozuelo Km2, 28220 Majadahonda-Madrid, SPAIN Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad Complutense de Madrid. Avda. Puerta de Hierro s/n, 28040 Madrid, SPAIN. [email protected] Keywords: Mediterranean, sandflies, species, cytb-nd1 Molecular methods are increasingly used for both species identification of sandflies and assessment of their population structure. In general they are based on DNA sequence analysis of targets previously amplified by polymerase chain reaction (PCR). However, this approach requires access to DNA sequence facilities and in some circumstances is time consuming. Though DNA sequencing provides the highest degree of information, other downstream applications of PCR are explored to help in species identification., It has thus been proposed that the amplification of a DNA region encompassing partially both the cytochrome-B (cytb) and the NADH dehydrogenase 1 (nd1) genes, and further digestion with the restriction enzyme AseI allows for the rapid identification of the most prevalent species of phlebotomine sandflies in the Mediterranean region based on their restriction fragment length polymorphism (RFLP). We sought to validate this methodology using sandflies collected from different regions in Spain, for which we amplified the cytb-nd1 target and obtained its DNA sequence. We completed this data set with DNA sequences available in the GenBank for species prevalent in the Mediterranean region. In all, 155 DNA sequences from 4 different phlebotomine sandfly species (Phlebotomus ariasi, P.papatasi, P. perniciosus, and Sergentomyia minuta) from Spain were generated for this work and added to 146 DNA sequences from 9 different phlebotomine species (P. ariasi, P. chabaudi, P. neglectus, P. papatasi, P. perfiliewi, P. perniciosus, P. riouxi, P. sergenti and S. minuta), from 18 different countries (Afghanistan, Algeria, Cyprus, Egypt, France, Iran, Israel, Italy, Jordan, Kosovo, Malta, Morocco, Palestine, Portugal, Spain, Syria, Tunisia, Turkey) obtained from the GenBank. In silico analysis revealed that this PCR-RFLP method does not provide a unique and specific profile for each species tested, intraspecific variability occurs in some of them (P. ariasi, P. papatasi, P. perniciosus and S. minuta), and there are patterns which are quite similar between different species (eg. P. chabaudi, P. riouxi, and P. sergenti show a RFLP pattern similar to intraspecific variants of P. ariasi and P. perniciosus). In our sample the results obtained by in silico analysis were further bench tested for confirmation. Though this method has previously been shown to be useful in Italy, it may not be applicable throughout theMediterranean region. Acknowledgement: COLCIENCIAS/COLFUTURO 2012. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 23-O Taxonomy & Phylogeny ID 24-O Taxonomy & Phylogeny MULTILOCUS REAL TIME PCR IDENTIFICATION OF PHLEBOTOMINE SAND FLIES CAPTURED IN SOUTHERN ITALY THE GENUS IDIOPHLEBOTOMUS (DIPTERA; PSYCHODIDAE) 34 Nicole Léger and Jérôme Depaquit Reale S, Torina S, Zaffora G, Cosenza M, Pitti R, D'Agostino R, Intile S, Piazza M, and Vitale F. Istituto zooprofilattico Sperimentale della Sicilia A. Mirri. Palermo Italy. Faculté de Pharmacie de Reims France. [email protected] [email protected] Keywords: Real Time PCR, Phlebotominae, Leishmania, Diptera Phlebotomine sand flies (Diptera: Psychodidae) living in Mediterranean area, are small-sized blood-sucking insects feeding on a wide range of hosts. They can be vectors of pathogens responsible for human and animal diseases as leishmaniasis. Out of over 800 sand fly species that have been described to date, approximately 10% are proven or suspected vectors of bacteria (e.g., Bartonella bacilliformis), viruses (e.g., Phlebovirus, Vesiculovirus) as well as protozoans from the genus Leishmania.. The identification of phlebotomine sand flies at species level relies on morphology of key structures including pharynx, spermathecae and cibarium for females and genitalia for males. Nonetheless, the morphological identification requires taxonomic expertise and it is time-consuming This study reports a combined analysis of ribosomal DNA target in phlebotomine sand flies from the Sicilian Mediterranean region. A 100 bp long fragments of DNA encompassing a part of ITS2 ribosomal region were amplified in Real Time test for Phlebotomus perniciosus, Phlebotomus perfiliewi, Phlebotomus neglectus, Phlebotomus papatasi, and Sergentomyia minuta, captured at various sites of southern Italy. Based on high interspecific rDNA difference, a universal couple of primers and 5 different probes were employed to reveal the percentage of the different sand fly species in the captures. Two different PCR mixtures were optimized to detect the cluster constituted by P. neglectus, P. perfiliewi, P. perniciosus, and the cluster P. papatasi, S. minuta respectively. On the basis of the obtained results, we conclude that the test can be used as preliminary approach to identify the species in captured pool of insects, and to individualize those that are considerably rare. This study demonstrates the utility of a multilocus approach to provide a tool for the molecular identification of the most prevalent phlebotomine sand flies in southern Europe. Obtained data showed the average percentage for each species in the catch, indicating the power to resolve the species mixture collected in the traps used in Leishmania endemic areas. All the species (16 or 15 ?) of Idiophlebotomus are localized inside the oriento-australasian area. This genera is the sister group of the African Spelaeophlebotomus , that is to say that they have a gondwanian origin and attests of an insular evolution of the phlebotomine sandflies during the long travel of the future India from Africa to Asia. Their morphological study is a key for the phylogeny of the Hertiginii tribe. ID 25-O Ecology & Epidemiology SAND FLY SPECIES TRANSMITTING VISCERAL LEISHMANIASIS AT THE SEMIARID REGION OF VENEZUELA Añez Néstor, Rojas Agustina, Yépez José Yancarlos Universidad de Los Andes, Mérida, Venezuela; Universidad Experimental Francisco de Miranda, Coro, Venezuela. [email protected] Keywords: Sand fly-species, Visceral-leishmaniasis, Semiarid, Venezuela A total of 12 sand fly species were recognized from 6000 specimens collected during a study carried out in 12 different localities at the semiarid region of western Venezuela where visceral leishmaniasis (VL) is endemic. From the total identified specimens, 81% (N=4892) belong to Lutzomyia longipalpis (15%; N=898) and Lu.evansi (66%; N=3994). The former, a species incriminated as vector of VL in most Latin-American countries, and Lu.evansi considered a potential vector of Leishmania infantum in VL endemic areas from Colombia and Venezuela. The study also revealed that Lu.evansi was present in all the 12 sampled localities (100%) while Lu.longipalpis was collected in 5 of them (41%). In addition, a PCR assay randomly performed to detect L.infantum from sand fly batches, collected in a village where active VL cases were detected, revealed infection in 3 of 5 (60%) and 4 of 18 (22%) batches of Lu.evansi and Lu.longipalpis, respectively. Statistical analysis using the Fisher Exact Test revealed no significant difference (Pvalue = 0.142) when results for Leishmania-infection in batches of both sand fly species were compared. These results suggest that despite the observed abundance and distribution of Lu.evansi over Lu.longipalpis, both sand fly species are sympatrically transmitting VL in the semiarid region of western Venezuela. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 26-O Ecology & Epidemiology ID 27-O Ecology & Epidemiology NATURAL BREEDING SITES OF LUTZOMYIA EVANSI (DIPTERA: PSYCHODIDAE) IN THE URBAN ZONE OF OVEJAS, SUCRE - COLOMBIA LUTZOMYIA FAUNAS (DIPTERA: PSYCHODIDAE) IN THE EAST SIDE OF THE ANDEAN CORDILLERA IN CUZCO, PERU Luis G. Estrada2, Horacio Cadena1, Edgar Ortega2, Luz A Acosta1, Andrés Vélez-Mira1, Rafael J Vivero1, Eduar E Bejarano2, Sandra Uribe3, Iván D Vélez1 Perez JE1, D Rado2, D Quispe2, A Quispe2, F Toccas2, M Chacon2, B Valencia1, A Llanos1, M Quispe2, E Aguilar2, R Pacheco2 & E Ogusuku3 1 PECET (Programa de Estudio y Control de Enfermedades Tropicales) - Universidad de Antioquia, Medellín, Colombia; 2Grupo de Investigaciones Biomédicas, Universidad de Sucre, Sincelejo, Colombia; 3Grupo de Investigación en Sistemática Molecular, Universidad Nacional, Medellín, Colombia. 35 1 Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia. 2Instituto Universitario de Enfermedades Tropicales y Biomedicina del Cuzco. Universidad Nacional San Antonio Abad de Cuzco, Peru. 3Direccion General de Salud Ambiental, Ministerio de Salud. Lima, Peru. [email protected] [email protected] Keywords: Peru, Lutzomyia, faunas, Cuzco Keywords: Immature, Phlebotomine, Leishmaniasis, Natural breeding The sand flies natural breeding places, despite their potential to assist the biological control, are poorly understood so far, due to the difficulties of isolation of immatures stages from the soil where they occur. The aim of this study was to identify natural breeding sites of Lutzomyia evansi in three zones of peri-urban area of the municipality of Ovejas, Sucre – Colombia. This region shows a bimodal climate: a rainy period from May to November and a dry period from December to April. The annual precipitation and mean temperature is 1380 mm and 28°C, respectively, and according to Holdridge the ecological lifezone is Tropical Dry Forest. The study was conducted between March 2013 and February 2014. Each zone consisted of 50 houses and search of breeding was carried out in the peri and intradomicile of these homes. Soil samples were taken around the bases of trees, leaf litter, hollow trees and soil under fallen trees. A total of approximately 503 kg of soil and associated organic matter were processed. Two different techniques: direct-observation and recovering emerging adults (incubation cages), were used to verify the presence of immature sand flies. During the rainy season the largest number of immature sand flies was recovered mainly in the base of trees. 70 natural breeding sites positive were identified and distributed as follows: zone1 n = 25; Zone 2 n =23; zone 3 n = 22. A total of 230 immature forms were found, which 135 (118 larvae and pupae 17) by direct-observation while 95 adults emerged by soil incubation. These results showed a richness of 6 species where Lutzomyia evansi was represented by 118 (51%) followed by Lu. rangeliana with 10 (4%), Lu. cayennensis cayennensis 7 (3%), Lu. micropyiga 6 (3%), Lu. atroclavata 4 (2%), and Lu. gomezi with 1 (0.4%). The 55.71% positive natural breeding sites were Lu. evansi, associated to the base of a shrub called locally as Uvito (Cordia dentata). 128 immature forms (including 44 exuviae recovered) that did not reach adulthood will be identified based on mitochondrial COI gene. This is the first record of peri-urban breeding sites of Lu. evansi in Colombia. The knowledge about natural breeding places for sand flies and vector density may represent useful information for directing efforts at biological control and consequently control the incidence of disease. Lutzomyia sandflies are wide distributed in the East side slopes of the Andean Cordillera of Cuzco Department. The highland vegetation is dominated by grasses and shrubs, the arboreal vegetation is mainly the introduced Eucalyptus spp. The lower lands are dominated by natural tropial forest. The whole area is endemic for bartonellosis (verruga peruana or Carrion disease) and leishmaniosis (uta).Objective: To determine the anthropophilic Lutzomyia fauna of the study area. Sandflies were collected with Shannon traps with protected human as attractant from 18:00 to 23:00 in the localities mentioned below. The Lutzomyia faunal composition is notoriously affected by the altitude, there is one species (L. (Helcocyrtomyia) peruensis) in Calca, Urubamba, Pisac, Ollantaytambo at around 3000 m of altitude; in San Pedro at 1380 m 5 species were reported (L. (H.) guderiani 74.9 %). These situations contrast with the reports for the lower lands, 26 species in Pillcopata at 1000 m (L. (Psychodopygus) c. carrerai 53.6 %); 37 species in Aguas Calientes at 890 m (L. (Nyssomyia) y. yuilli 57%); 34 species in Alto Ivochote at 880 m ((L. (Nyssomyia) y. yuilli 53%); and 29 species in Paratori at 450 m (L.(P.) llanosmartinsi 72.5 %). Lutzomyia (Helcocyrtomyia) species predominate in the highest altitudes (>3000m) along the Andean Cordillera in Peru, this is the case of L. peruensis in Cuzco. The west side of the Cordillera is dry and arboreal vegetation is restricted to protected areas in the bottom of some valleys. Lutzomyia fauna in this area is 3 to 6 species, Lutzomyia peruensis and L verrucarum are the most common species, both are vectors of bartonellosis and Andean leishmaniosis (uta). N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 28-O Ecology & Epidemiology ID 29-O Ecology & Epidemiology DIFFERENTIAL EXPOSURE TO SAND FLY BITES IN SAUDI ARABIA PREDICTING THE GEOGRAPHICAL DISTRIBUTION OF LEISHMANIASIS VECTOR SPECIES IN CENTRAL WEST BRAZIL Karina Mondragon-Shem1; Waleed Alsalem1,2; Louise Kelly-Hope1,3; Maha Abdeladhim4; Mohamed Alzahrani2; Ziad Memish2; Jesus Valenzuela4; Alvaro Acosta-Serrano1, 5 1 Department of Parasitology, Liverpool School of Tropical Medicine, England, UK; Saudi Ministry of Health, Riyadh, Kingdom of Saudi Arabia; 3Centre for Neglected Tropical Diseases, Liverpool School of Tropical Medicine, England, UK; 4Vector Molecular Biology Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, Maryland, United States of America; 5 Department of Vector Biology, Liverpool School of Tropical Medicine, England, UK. 2 [email protected] 36 Paulo Silva de Almeida, Alan Sciamarelli, Josué Raizer, Jaqueline Aparecida Menegatti, Sandra Cristina Negreli Moreira Hermes, Maria do Socorro Laurentino de Carvalho, Andrey José de Andrade, Rodrigo Gurgel-Gonçalves Universidade de Brasília. [email protected] Keywords: Phlebotominae, ecologic niche modeling, Central West Brazil, surveillance. Keywords: Phlebotomus papatasi, biting exposure. Human antibody response to sandfly saliva can serve as a marker of exposure to the bite of vector species, and is valuable to assess risk of disease in different endemic settings. In Saudi Arabia, zoonotic cutaneous leishmaniasis (ZCL) is caused by Leishmania major and transmitted by Phlebotomus papatasi, while anthroponotic CL (ACL) is caused by L. tropica and vectored by Ph. sergenti. Salivary protein SP32 of Ph. papatasi, the main target of the human immune response against the saliva of this species, is an ideal biomarker to measure exposure. The recombinant form of this protein, rPpSP32, allows for mass screening of sera in endemic areas. In this work, we determined the level of exposure to Ph. papatasi bites in individuals from CL endemic areas in Saudi Arabia. We obtained sera from healthy residents CL as well as patients with active and cured infection, from CL foci in Al-Ahsa, Al-Madinah and Asir. We measured the levels of anti-SP32 antibodies by ELISA using rPpSP32. Additionally, we collected sandflies from the same regions to identify the vector species present the foci. rPpSP32 was successfully recognized by human sera from the ZCL endemic areas in Saudi Arabia. We found higher titres of anti-SP32 antibodies in CL patients than in healthy and CL-cured individuals in Al-Hasa, while in Al-Madinah there was no difference between the groups. In both of these ZCL foci Ph. papatasi was the predominant sand fly species. Interestingly, we also detected a significant difference between the levels of anti-SP32 antibodies in patients with papular and nodular lesion types in Al-Madinah. In Asir, where Ph. sergenti was the most common species, we found low levels of anti-SP32 antibodies in ACL patients. This revealed a potential cross-reaction with the Ph. sergenti SP44 PpSP32-like salivary protein, which was supported by the high similarity with the SP32 protein sequence. Additionally, we discovered the non-local workforce in Al Ahsa exhibited higher antibody levels and increased lesion numbers compared to long-term residents, which will have important implications for leishmaniasis control programmes in Saudi Arabia. This is the first study using rPpSP32 protein to assess biting exposure in CL patients. Cross-reaction with Ph. sergenti salivary proteins must be kept in mind when evaluating exposure in areas where both species co-exist. Our results suggest a correlation between biting exposure, disease outcome and clinical presentation. Central West Region of Brazil (CWB) comprises the states of Mato Grosso (MT), Mato Grosso do Sul (MS), Goiás (GO) and the Federal District (FD). The present study analyzed the geographical distribution of the main sand fly species associated with leishmaniasis in CWB and climatic factors associated with its occurrence. Distributional data of seven sand fly species were obtained by the entomology laboratories of “Secretarias Estaduais de Saúde”, museums and literature review between 1978 and 2014. Data were separated into two sets: one for model calibration (75% of points) and one for model evaluation (25% of points). Climatic data covered areas of CWB, resample to a spatial resolution of 5 km. Models were produced based on eight climatic variables using Maxent. Median output grids were used as a hypothesis of suitability, and they were imported into ArcView and DIVA for analysis. Model accuracy was assessed by examining omission rate associated with test points. Maxen's jackknife test was used to identify variables that most influenced models. In total 566 records were analyzed. Bichromomyia flaviscutellata (n=61), Lutzomyia longipalpis (n=128) and Nyssomyia whitmani (n=200) had wide distribution in CWB. Lu. cruzi occurred in 52 municipalities in southwestern and eastern regions of MT, northern MS and southern GO. Ny. antunesi showed more restricted distribution in the state of MT and north of MS (n=71). Ny. intermedia (n=33) and Ny. neivai (n=21) were distributed in the states of GO, MT and FD. Predictive models indicated areas with high climatic suitability for Ny. whitmani (southern and northern regions of MT and MS, central GO), Lu. longipalpis (southwestern MS and northern MT), Lu. cruzi (southeastern MT, southwestern GO and northern MS), Bi. flaviscutellata (northern MT), Ny. intermedia (central GO and southwestern MT), Ny. neivai (southern, southwestern and eastern MS), and Ny. antunesi (central MT and eastern GO). The models showed < 1% omission error. The variable that most influenced models was the temperature in the coldest months. Moreover, the occurrence of Lu. longipalpis was associated with dry areas, unlike the models of the other species that were strongly influenced by the annual precipitation. The results demonstrated different patterns of geographical distributions for leishmaniasis vectors in the CWB, which can assist surveillance and control strategies of this disease. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 37 ID 30-O Ecology & Epidemiology ID 31-O Ecology & Epidemiology ECOLOGY OF SANDFLIES IN RISK ZONES FOR TEGUMENTARY LEISHMANIASIS TRANSMISSION IN SANTARÉM MUNICIPALITY, WESTERN PARÁ STATE, BRAZIL PHLEBOTOMUS ORIENTALIS AND THE FALL AND RISE IN VISCERAL LEISHMANIASIS IN EAST AFRICA Dia-Eldin A Elnaiem Raquel Gonçalves, Daniela Cristina Soares, Ricardo Guimarães, Walter Souza Santos, Gilberto Cesar de Sousa, Anadeiva Portela Chagas, Lourdes Maria Garcez University of Maryland Eastern Shore. [email protected] Instituto Evandro Chagas/SVS/MS and Universidade do Estado do Pará. [email protected] Keywords: Phlebotmus orientalis , East Africa Keywords: Ecology, sand flies, leishmaniasis, risk zone Phlebotomus orientalis sand fly is the main vector of Leishmania donovani in most foci of visceral leishmaniasis (VL, kala azar) in East Africa. The two other vectors, P. martini and P. celiae, are responsible of transmission of L. donovani in limited foci in the southeastern region of Ethiopia and the border areas of the Republic of South Sudan (RSS) and Kenya. Due to marked fluctuations in the abundance of P. orientalis, East African visceral leishmaniasis is characterized by periodic outbreaks that result in high morbidity and mortality rates. The severity of these outbreaks is exacerbated by lack of knowledge that allows their prediction both in time and space. Our observations in several areas in Sudan indicate that this vector may disappear for several years and then erupt in large numbers. While we believe the cyclical nature of VL and its vector is most likely related to climatic conditions, little is known about the specific environmental factors responsible for the epidemics. This is partly due to lack of reliable surveillance system that provide accurate estimates of number of cases and abundance of the vector. We recently embarked on a multicenter research initiative to elucidate the environmental determinants affecting the fall and rise of VL and use them to construct a model that can help predict future outbreaks. We correlated numbers of VL cases with climatic data in eastern Sudan and RSS. Our results showed that the incidence of the disease is negatively correlated with the rainfall at the end of the dry season. These findings correspond closely with our observations on the ecology of P. orientalis. Future studies will attempt to build Early Warning System for Visceral Leishmaniasis, based on climate and other environmental factors. Tegumentary leishmaniasis (TL) in Brazil used to be near 10 times more incident than visceral leishmaniasis (VL). The environmental and socio-economic context influence both in the different endemic areas. Pará is among the five Brazilian states with the highest incidence rates of TL over the last years. The municipality of Santarém is one of the main centres reporting new cases of the disease. We studied the ecology of sand flies in two spatial circuits (SCs) of TL in Santarém using the program PAST v.2.03 (PAlentological STatistic) for calculating ecological indexes: Shannon (H'), Pielou (J) and Dominance (D). To define the SCs we used geographical coordinates from transmission and housing locals of patients and the Kernel density estimator. Thirty-one records selected of a database with 102 TL patients reported in 2010 and 2011 were included. The patients lived in plateau areas from Santarém, had one or more primary cutaneous/mucocutaneous lesion and the TL confirmed by parasitological and molecular diagnosis. Among the etiologic agents infecting the patients there were Leishmania (Viannia) braziliensis, L. (V.) shawi, L. (V.) lainsoni, L. (V.) naiffi and one hybrid form of L. (V.) braziliensis/guyanensis. We identified two SCs of TL in images classified according the deforestation (2002-2009) and the land uses (Basin Restoration Program, PRODES – TerraClass Project; National Institute of Space Research, INPE, Brazil, 2011): Jatobá and CortaCorda. There were captured 417 sand flies (24 species) throughout two entomological surveys using Shannon and CDC traps. Most were females (312; 75%) and the sand flies number did not vary in wet (226/417; 54%) and dry (191/417; 46%) seasons. Nine genus were identified: Bichromomyia, Evandromyia, Lutzomyia, Micropygomyia, Nyssomyia, Psathyromyia, Psychodopygus, Sciopemyia, Trichopygomyia. The ecological indexes varied (H'=0.5-1.72; J= 0.44-0.78; D=0.46-0.91) and confirmed high diversity and the importance of the vectors Lutzomyia longipalpis (VL) and Psychodopygus complexus (TL) in Jatobá (urban-rural and ecotourism area) and Psychodopygus davisi (TL) in Corta-Corda (rural farm, around a hydroelectric dam). There were less important the vectors Nyssomyia antunesi, Bichromomyia flaviscutellata and Nyssomyia anduzei (TL). The results support the Leishmania species diversity in the patients living in the SCs of TL, representing the importance of such analysis in entomological surveys. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 32-O Ecology & Epidemiology ID 33-O Ecology & Epidemiology DISTRIBUTION OF LEISHMANIASIS VECTORS IN THE MEDITERRANEAN BASIN IN THE LIGHT OF CLIMATE CHANGE CHARACTERIZATION OF BREEDING SITES OF PHLEBOTOMINE SAND FLIES IN THE JUDEAN DESERT AND NORTH-WEST ETHIOPIA Bilel Chalghaf, Amine Toumi, Sadok Chlif, Jihene Bettaieb, Mariem Harrabi, GozeBertin Bénié, Afif Ben Salah Aviad Moncaz, Oscar Kirstein, Araya Gebresellassie, Wossenseged Lemma, Solomon Yared, Teshome Gebre-Michael, Asrat Hailu, Moshe Shenker and Alon Warburg Institut Pasteur Tunis, Universite de Sherbrooke. [email protected] Keywords: climate change, ecological niche, modeling, sand flies Due to climate change, the geographical distribution of sandflies during the last decades has shifted northward from latitudes below 45○N in south Europe (Martens and McMichael 2002) to latitudes just above 50○N (Naucke et al., 2008). Recent studies show that some phlebotomine sandflies were recorded in several parts of Germany and Belgium (Naucke et al., 2008, Mencke; 2011). In Central Europe, some autochthone leishmaniasis cases are being recorded in regions traditionally regarded as leishmaniasis-free countries (Gogoaşe et al., 2013, Tánczos et al., 2012).In this study we attempted to predict current distribution of six leishmaniasis vectors in the Mediterranean basin and forecast species geographical shift under future climate scenarios using ensemble ecological niche modeling approach. Species records were obtained from scientific surveys published in the research literature between 2006 and 2012. A series of climate metrics, describing temperature and precipitation in the study area under two climatic scenarios, were obtained from WorldClim database. We used six individual models contributing to the consensus model including two regression techniques (Generalized Linear Models, GLM; Generalized Additive Models, GAM), two machine learning techniques (Artificial Neural Networks, ANNs; Maximum Entropy, MaxEnt) and two classification techniques (Surface Range Envelop, SRE; Random Forest, RF) (why two models from each category of models?). Alternative formulation: A consensus model was derived from six varieties of modeling approaches (regression, machine learning and classification techniques) in order to ensure valid prediction of distribution of vectors under different climate scenarios. Model performance was generally high for the included species with a specificity (True Negative Rate) ranging from 81.03% to 96.52% (mean=86.94%) and a sensitivity (True Positive Rate) ranging from 87.93% to 100% (mean=96.98%). Our work evidenced the hypothesis of the wide spread of Leishamania vectors under climate change scenarios. All the studied species are prospected to gain new areas that are actually not suitable for vectors' survival. Phlebotomine sandflies are prospected to invade extra-Mediterranean regions especially West. 38 The Kuvin Center for the Study of Infectious and Tropical Diseases, Department of Microbiology and Molecular Genetics, The Institute of Medical Research IsraelCanada, Faculty of Medicine, The Hebrew University of Jerusalem, Hadassah Medical School, Jerusalem, Israel. [email protected] Keywords: Phlebotomus vertisols caves trees Identification and characterization of breeding sites is essential for the implementation of source-reduction for the control of sand flies and leishmaniasis. Sand fly breeding sites were identified and characterized in two foci of leishmaniasis: In the Judean Desert where the dominant sand fly species that transmits Leishmania tropica is Phlebotomus sergenti, and in NW Ethiopia where L. donovani is transmitted by Ph. orientalis. Sand flies emerging from their breeding sites were trapped by emergence traps (ETs) for 2–8 consecutive nights. In the Judean desert, Ph. sergenti were trapped emerging from caves and artificial support walls made of large rocks. . Young (recently eclosed) male sand flies were found in both habitats. The temperatures and relative humidity inside caves were stable and soil samples collected in the cave opening were rich in organic matter. In Ethiopia Ph. orientalis breeding sites were identified in cracks in vertisols including fallow fields, dry riverbed/bank and near trees. No Phlebotomus spp. were trapped in sandy clay loam soil. More Ph. orientalis were trapped near tree trunks than in open fields but the difference was insignificant. Large numbers of Sergentomyia spp. were trapped along with Ph. orientalis mostly outnumbering Phlebotomus spp. by an order of magnitude. Initial microclimatic monitoring in the two habitats was conducted at the end of rainy season, and showed stable subterranean temperatures and RH. Conditions remained stable through early dry season close to trees but in open fields temperature fluctuated by 5-6ºC and RH decreased significantly. Hypoxic conditions evolved during in tree root systems during the rainy season but oxygen levels rebounded to atmospheric values once cracks reformed during the dry season. Organic matter content was high (>5%) in both habitats. Soil pH was moderately alkaline and electrical conductivity was low in both habitats and depths. Low oxygen concentrations in soil close to tree root systems during the rainy season may be caused by microbial metabolism. Decomposing organic matter, stable temperatures and elevated soil moisture maintained throughout dry season near the trees at shallow and deep horizons, and in the open field at deep horizon, probably maintain suitable conditions for sand fly larvae. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 34-O Ecology & Epidemiology ID 35-O Ecology & Epidemiology F I N E S C A L E A N A LY S E S O F FA C T O R S CONTRIBUTING TO THE TRANSMISSION OF VISCERAL LEISHMANIASIS IN NORTH ETHIOPIAN VILLAGES: AN ECO-EPIDEMIOLOGICAL APPROACH SERO-PREVALENCE L. DONOVANI IN LABOUR MIGRANTS POPULATION AND ENTOMOLOGICAL RISK FACTORS IN EXTRA-DOMESTIC HABITATS OF METEMA-HUMERA LOWLANDS - KALA-AZAR ENDEMIC AREAS IN THE NORTHWEST ETHIOPIA Oscar David Kirstein, Araya Gebreselassie, Aviad Moncaz, Asrat Hailu, Teshome Gebre-Michael and Alon Warburg Wossenseged Lemma1,2, Habte Tekie2, Meshesha Balkew3, Teshome Gebre-Michael3, Alon Warburg4, Asrat Hailu5 Department of Microbiology & Molecular Genetics, The Institute for Medical Research Israel-Canada, The Kuvin Centre for the Study of Infectious &Tropical Diseases, The Hebrew University – Hadassah Medical School, The Hebrew University of Jerusalem. [email protected] Keywords: Sand fly, Kala Azar, Epidemiology, Visceral leishmaniasis Landscape ecology and landscape epidemiology comprise a set of tools used for spatial and temporal analyses of vector-borne and other diseases. In Sudan and Northern Ethiopia, visceral leishmaniasis (VL) caused by Leishmania donovani, is transmitted by Phlebotomus orientalis. The epidemiology of VL is the result of a complex matrix of interactions between the vectors, the pathogens and the hosts with the environment. This study aims to characterize the risk factors for contracting VL by gaining a profound understanding of the factors contributing to its transmission to humans. During 2010 - 2013 a transverse cohort study was conducted, based on a census comprising over 11,000 individuals in 18 villages in the Tahtay Adiabo district of northern Ethiopia. Around 4,900 individuals were screened and tested for infection with L. donovani. Based on the incidence of VL reported during the census, the Leishmania infection rates determined by PCR during the cohort samplings and the environmental and geographical variables, three villages were selected for an in-depth ecoepidemiological study. In 2014 a nested case-cohort study of ~400 individuals was conducted in the three selected villages, confounder information available in the previous cohort data-base was used to select controls that closely match cases in term of sex, age and geographical location. By analyzing satellite images taken during the dry season, it was possible to identify that the majority of villages affected by VL during recent years are inside an area characterized by vertisols (radius 28 Km). Remote sensing analysis showed high spectral variability produced by a mix of vertisols, sandstone outcrops, other sedimentary rocks and varied vegetation land cover. Entomological studies identified that P. orientalis is closely associated with vertisol ecosystems rather than specific micro-environments, being most abundant in cracked black cotton soils. Spatial analysis of the incidence of VL, show a clustered distribution within populated areas and micro-clusters of more than one case per house or nearby house. Houses of patients were most frequently within a few meters of vertisol fields. These observations may support the hypothesis that the infection with L. donovani occurs in domestic environments rather than in the field. The current study is ongoing and more data analysis will validate or otherwise affect the results presented here. 39 1 Department of Parasitology, School of Biomedical and Laboratory Sciences, College of Medicine and Health Sciences, University of Gondar, Gondar, Ethiopia. 2 Department of zoology, Faculty of Life Science, Addis Ababa University, Addis Ababa, Ethiopia. 3Aklilu Lemma Institute of Pathobiology, Addis Ababa University, Addis Ababa, Ethiopia. 4Department of Microbiology & Molecular Genetics, The Institute for Medical Research Israel-Canada, The Kuvin Centre for the Study of Infectious & Tropical Diseases, The Hebrew University – Hadassah Medical School, The Hebrew University of Jerusalem, 91120, Israel. 5Department of Microbiology, Immunology & Parasitology, Faculty of Medicine, Addis Ababa University, Addis Ababa, Ethiopia. [email protected] Keywords: kala-azar infection, labour migrants, risk factors; Ethiopia Visceral leishmaniasis (VL) or kala-azar is a systemic disease transmitted by phlebotomine sand flies and almost always fatal if left untreated. The aim this study was to evaluate sero prevalence of kalaazar infection and associated risk factors which affect the incidence of kala-azar. Study on seasonal population dynamics of P.orientalis were conducted in different villages of Humera before blood samples was randomly obtained from labour migrants involved in harvest of sesame for Direct Agglutination Test (DAT) after oral consents were obtained during October 27 to November 7, 2013. A structured questionnaire was used to obtain demographic information and factors that affect the incidence of kala-azar infection in labour migrants. About 80% of the labour migrants were aged from 15 to 34 years and came from Amhara (75.2%), Tigray (21.2%) and other parts of Ethiopia (3.6%). Stayed both harvest and weed season in Humera, Involved in night harvest, Sleep in the farm under Balanites aegiptica and Knowledge about, at least, a sign or a symptom of kala-azar were the major factors (P< 0.05) influencing kala-azar infection (DAT positivity or sero reaction). The total P. orientalis of 7022 (3277 female and 3745 male) were collected from agricultural fields and thickets in A. seyal in Adebay, Rawyal, Gelanzeraf and Mysegen – Mehari using 932 CDC traps. The month with the highest mean P. orientalis/CDC (20.47) abundance was in April. The mean P. orientalis/CDC during dry season (Nov - April) was highest (Mean ± Std D =7.33 ± 18.11) compared to May – August weeding (2.05 ± 4.47) and September – October sesame harvest (0.41± 0.77) seasons. Kala-azar infection in labour migrants seemed to relate with P. orientalis bite during May – August weeding season compared to September – October least P. orientalis abundance harvest season. Peak Kala-azar incidence in permanently settled farmers in villages of Shiraro also seemed related to P. orientalis bite during May – Augest weeding season which took 2- 6 months before the peak manifestations of the disease in January. Labour migrants are exposed to kala-azar infection during May - August clearing farm and weeding season. But, stay during September – October harvest season or involve in night harvest were not increase the risk of kala-azar infection as P. orientalis population drops to almost to zero. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 40 ID 36-O Ecology & Epidemiology ID 37-O Ecology & Epidemiology HOST PREFERENCE PATTERNS OF PHLEBOTOMUS (LARROUSSIUS) ORIENTALIS PARROT, 1936 (DIPTERA: PSYCHODIDAE) IN THE VISCERAL LEISHMANIASIS ENDEMIC AREA OF TAHTAY ADIYABO DISTRICT, NORTHERN ETHIOPIA BIONOMICS OF PHLEBOTOMINE SANDFLIES IN A HIGHLAND KALA-AZAR FOCUS IN LIBO-KEMKEM DISTRICT, NORTHWESTERN ETHIOPIA Esayas Aklilu, Habte Tekie, Meshesha Balkew, Alon Warburg, Asrat Hailu, Teshome Gebre-Michael Araya Gebresilassie1, Ibrahim Abbasi2, Oscar David Kirstein2, Aviad Moncaz2, Habte Tekie1, Meshesha Balkew3, Alon Warburg2, Asrat Hailu4, and Teshome Gebre-Michael3 Addis Ababa University. [email protected] 1 Department of Zoological Sciences, Addis Ababa University, Addis Ababa, Ethiopia, Ethiopia; 2Department of Microbiology and Molecular Genetics, The Institute of Medical Research Israel-Canada The Kuvin Center for the Study of Infectious and Tropical Diseases, Faculty of Medicine, The Hebrew University, Hadassah Medical School, Jerusalem, Israel; 3Aklilu Lemma Institute of Pathobiology, Addis Ababa University, Addis Ababa, Ethiopia; 4Department of Microbiology, Immunology and Parasitology, Faculty of Medicine, Addis Ababa University, Addis Ababa, Ethiopia. [email protected] Keywords: Bloodmeals, P. orientalis, host preference, Kala-azar Host choice and feeding success of sand flies are important factors for understanding the epidemiology and for developing efficient control strategies for visceral leishmaniasis. The aim of the study was to determine the host preference patterns of Phlebotomus orientalis from Tahtay Adiyabo district, Northern Ethiopia. Host attractiveness experiments were conducted by presenting wild sand flies with a choice of seven baits (human, cattle, sheep, goat, donkey, dog and chicken) and control (without bait) in locally designed tent traps. For blood meal analysis, sand flies were collected from three villages using CDC light and sticky traps. DNA was extracted from the blood fed sand flies and PCR amplified for vertebrate-specific cytochrome (cyt) b region, followed by reverse-line blot (RLB) analysis. Attraction and feeding success of P. orientalis indicated significant differences (P < 0.005) between baits. Abundance of P. orientalis on traps baited with cattle was higher than other animal baits and controls. Female sand flies were significantly more attracted to donkey and cattle than human, dog, goat, sheep or chicken-baited traps. P. orientalis females engorged more frequently on donkey and cows than they did on other hosts (goat, sheep, dog, and chicken). Out of 180 P. orientalis tested for the source of bloodmeals by cyt b PCR and RLB, the blood meals of 135 were successfully identified. P. orientalis mainly fed on bovine followed by human, goat, sheep and camel. Mixed blood feeding was also identified in ten female P. orientalis. The epidemiological significance of these domestic animals as blood sources for P. orientalis and possible reservoir hosts of L. donovani should be thoroughly investigated for better understanding the transmission dynamics of visceral leishmaniasis in northern Ethiopia. Keywords: Visceral leishmaniasis, Phlebotomus orientalis, highland, Ethiopia The bionomics of phlebotomine sandflies (Diptera: Psychodidae) were studied for one year from May 2011 to April 2012 in three villages of Libo-Kemkem district, a highland area where visceral leishmaniasis has become a major public health problem. The aim of the present study was to elucidate species composition, resting habits, seasonal fluctuations of phlebotomine sandflies and incrimination of the vector. CDC light trap, sticky trap and knockdown space spray were used for collection of sandflies. Dissection and molecular approaches were used for Leishmania detection from female Phlebotomus that were collected by using CDC light traps. A total of 10, 776 sandflies comprising of two species of the genus Phlebotomus and five species of the genus Sergentomyia were collected. Phlebotomus orientalis was the predominant species accounting 86.6% of the total sandfly captured. The remaining species in descending order were Sergentomyia bedfordi group, S. squamipleuris, S. schwetzi, S. africana, S. clydei and P. rodhaini. Sandflies were present in most part of the year, except during the main rainy season (July-September). Phlebotomus orientalis exhibited two peaks of density, a smaller in January 2012 (6.5flies per trap night for CDC light trap; 0.98flies/m2/night for STs) and the larger one in March for STs (1.06flies/m2/night) and April for CDC light trap (18. 7flies per trap night). Abundance of the flies were positively correlated with average temperature (r=0.53 for CDC; r=0.563 for STs) and negatively correlated with rainfall (r= -0.272 for CDC; r= 0.171 for STs). The male/female ratio of P. orientalis in the total captures was 1.87:1. Behaviorally, P. orientalis exhibited both exophilic and exophagic. A total of 1060 P. orientalis and 1 P. rodhaini dissected were found to be negative by microscopic examination. A similar result was obtained when 247 P. orientalis females were processed by using Polymerase chain reaction for detection of the parasite. Although natural infection was not detected in the current study, P. orientalis is seems to be a vector of VL in this highland area of the country mainly due to its densely populations and other circumstantial evidences. Therefore, control of the disease in this particular area should involve designing of tactics that mainly target the vector of the disease by considering its seasonal abundance and behaviors. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 38-O Ecology & Epidemiology ID 39-O Ecology & Epidemiology SPECIES COMPOSITION, ABUNDANCE AND SEASONAL DYNAMICS OF PHLEBOTOMUS SPECIES IN A VISCERAL LEISHMANIASIS ENDEMIC AREA OF NORTHWEST ETHIOPIA PHLEBOTOMINE SAND FLIES CROSSING A N AT O L I A : D O B O R D E R S L I M I T T H E I R DISTRIBUTION? Solomon Yared, Meshesha Balkew, Alon Warburg, Asrat Hailu and Teshome Gebre-Michael O Kasap Erisoz1, A Belen2, C Alkan2, F Gunay1, V Dvorak3, K Ergunay4, S Aydın5, J Votypka3, A-L Banuls6, R Charrel2, A Özkul7, Y Özbel8, P Volf3, B Alten1 Addis Ababa University. [email protected] Keywords: Visceral leishmaniasis, Phlebotomus spp, P.orientalis, Habitat preference Visceral leishmaniasis (VL) is significant public health problem in northwest Ethiopia particularly in Kafta Humera district. This study was conceived to investigate the species composition and population dynamics of sandflies (altitude 500-600masl) in five urban and semiurban area of Kafta Humera district namely Setit Humera, Mykadra, Rawiyan, Bereket and Adebay. Sandflies were collected for three nights monthly from May 2011 to April 2012 using CDC light-traps and sticky traps. Traps were placed within villages, at periphery of villages and farm fields. Sticky traps were also used for sampling indoor active sandflies. A total of 13097 sand flies comprising of six Phlebotomus species of four subgenera were indentified: Phlebotomus (Larroussius) orientalis, P.(Phlebotomus) papatasi, P.(Ph.) bergeroti, P.(Ph.) duboscqi, P. (Paraphlebotomus) alexanderi, and P. (Anaphlebotomus) rodhani. In addition, two Parvidens (P. lesleyae and P.heischi) species were recorded in the study area. Among these, P. orientalis was the most predominant species, accounting for (58.12%) followed by P. papatasi (29.62%), P. lesleyae (5.61%), P. bergeroti (3.80%), and P. duboscqi (2.07 %), P. alexandri (0.37%), P. heischi (0.24%) and P. rodhaini (0.18 %). On average, significantly higher densities of P. orientalis were caught in the periurban area of Adebay with compared to the urban area of Setit Humera. Overall, 684 Phlebotomus females were dissected for detection of Leishmania infection, but none was infected. Significant positive correlation was found between the monthly abundance of P. orinetalis and P. papatasi and the monthly averages temperature (P<0.05) whereas negatively correlated with monthly average rainfall and relative humidity (P<0.05). Abundance of P. orientalis and P. papatasi significantly increased during the dry season (January- May) but decreased in the wet season (June - September) (P<0.05) in the study areas. With regard to habitat preferences, a large number of P.orientalis was collected from outdoors especially in the farm fields with cracked vertisol. In contrast, P. papatasi was highly abundant indoors as well as outdoors within and the periphery of villages. P. orientalis is a predominant species and highly abundant during the dry season and possibly able to transmit VL in the study area. Appropriate control methods should be designed according to the knowledge of P. orientalis habitat preferences and seasonal dynamics in relation to climate condition. 41 1 Faculty of Science, Hacettepe University, Department of Biology, Ecology Section, Beytepe-Ankara-Turkey; 2Unite des Virus Emergents, Marseille Universite, UMR190 Emergence des Pathologies Virales, Faculte de Medecine, Marseille, France; 3 Parasitology Department, Charles University, Faculty of Science, Prague, Czech Republic; 4Department of Medical Microbiology, Hacettepe University, Faculty of Medicine, Virology Unit, Sihhiye-Ankara-Turkey; 5Department of Communication Sciences, Hacettepe University, Faculty of Communication, Beytepe-Ankara-Turkey; 6 IRD (IRD 224-CNRS 5290-UM1-UM2), MIVEGEC, Montpellier, France; 7 Department of Virology, Ankara University, Faculty of Veterinary Medicine, DiskapiAnkara-Turkey; 8Parasitology Department, Ege University, Faculty of Medicine, Bornova, Izmir, Turkey. [email protected] Keywords: Sandflies, ecology, epidemiology, Anatolia The term of phlebotomine sand fly was used by Philippo Bonanni in 1691 and then first description was made by Scopoli, under the name of Bibio papatasi. The name of genus, Phlebotomus, was given in 1840 by Rodani and Berte. Since then, more than 800 species belonging different genera and subgenera were identified in all over the world. Their vectorial role for transmitting Leishmania parasites and phleboviruses were defined at the beginning and mid of the 20th century, respectively. The Western Palearctic (WP) is composed of Europe, Middle East and North Africa. In this territory, the Mediterranean Sea, and the land under the influence of the Mediterranean Sea is the most important geographical character for both migration and dispersion of organisms; especially for invertebrates including sand flies together with pathogen organisms in it. Anatolia (Asia-Minor) takes place on the crossroads of this area and has a role as a bridge geographically and biologically. Leishmaniasis and arbovirus infections due to phleboviruses have wide geographical distributions in the Western Palearctic, and give rise to occasional epidemic outbreaks. In numerous countries, increasing risk factors are making sand fly-borne diseases a major public and veterinary health problem. Many studies on phylogenetic relationship among sand fly taxa, their distribution, population structure and diseases of phlebotomine species have been already published, but there are still many gaps waiting to be filled up in, especially, Anatolia. In this point, scientists have to discuss some deficiencies under cover of geography, history and phylogenetic studies to understand the mechanisms of distribution of both sand fly species and their pathogens in Anatolia. In this presentation, updates in the distribution of sand fly species with state of art maps of EUVBORNET project, possible new species, leishmaniasis and phleboviruses epidemiology will be discussed with an emphasis on several studies performed by our group between 2000 and present in Anatolia. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 42 ID 40-O Ecology & Epidemiology ID 41-O EPIDEMIOLOGICAL STUDY ON SAND FLIES IN AN ENDEMIC FOCUS OF CUTANEOUS LEISHMANIASIS, BUSHEHR CITY IN SOUTHWESTERN IRAN ECO-EPIDEMIOLOGY OF LEISHMANIASES: VECTORS AND NEW SCENARIOS OF TRANSMISSION. M R Yaghoobi-Ershadi1, M Darvishi1, F Shahbazi1, AA Akhavan1, R Jafari2, M Khajeian3, H Soleimani4, H Darabi3, M H Arandian2 Jeffrey Jon Shaw Biomedical Sciences Institute - University of São Paulo, São Paulo - SP – Brasil School of Public Health, Tehran University of Medical Sciences, P.O.Box 644614155Tehran, Iran [email protected] Keywords: Epidemiology, P. sergenti, L. turanica, Iran Cutaneous Leishmaniasis (CL) is the most important health problem in the city of Bushehr, southwestern Iran. The objective of the study was to determine some ecological aspects of sand flies in the city during 2010-2011. Sand flies were collected monthly from outdoors and indoors by sticky traps at four selected districts of the city. They were also dissected and examined by Nested-PCR for identification of the parasite during August- September of 2011. A total of 1234 adult sand flies were collected and 6 species including 3 of Genus Phlebotomus and 3 of Genus Sergentomyia were identified. Four species including P. papatasi (3.98 %), P. sergenti (1.14 %), S. tiberiadis (87.18 %) and S. baghdadis (7.7 %) were found indoors. Five species including P. papatasi (3.47 %), P. sergenti (3.17 %), P. alexandri (0.1%), S. tiberiadis (77.85 %) and S. baghdadis (15.41 %) were collected from outdoors. Sand flies started to appear from March and disappear at the end of January. There was only one peak in the density curve in July. The study revealed that S. tiberiadis and S. baghdadis could enter indoors which 89 % and 81.8 % of them were found blood-fed respectively. Moreover, P. papatasi, S. tiberiadis and S. baghdadis were active indoors and outdoors in most months of the year. Nested-PCR of P. papatasi females were positive against kinetoplast DNA of L. major and L. turanica and also mixed natural infections were found by L. gerbilli and L. turanica. Moreover mixed infections by L. major and L. turanica were observed in this species. Phlebotomus sergenti females were found infected with DNA of L. turanica and this the first report of natural infection and detection of the parasite from this sand fly species in worldwide. We have no idea of the number of vectors involved in the transmission of the leishmaniases but in 2010 a WHO publication lists 91 as suspected or proven. A major difficulty is how to distinguish these two categories. Attempts to rationalize vector status have been made by using a variety of epidemiological and parasitological criteria. The problem is that it is difficult to obtain all these criteria. The following are those that it is quite feasible to attain: confirmation that the parasite found in the fly is the same as that found man or suspected reservoirs; the intensity and localization of the infection; concomitant ecological distribution with the disease; the degree of anthropophily and zoophily. It is also important to show that fly transmits the parasite. During recent years the success rate of experimental transmissions has improved tremendously but some vectors are refractory to colonization. This can be overcome by using wild caught flies. It does not matter whether or not the infections are experimental or natural. It is against this background that we have to assess vectors and changes in transmission patterns. Both the cutaneous and visceral leishmaniases are zoonoses with the exception of Indian kalazar. Man's relationship to the ecological niches in which the enzootic cycle occurs is a dynamic process that depends on a multiple of factors linked to sandfly ecology. Studies on the ecology of the different vectors are fundamental in understanding new transmission scenarios. The major changes in transmission are associated with modifications of ecological niches that are related to either man's activities or natural phenomena or a combination of both. The challenge is to know whether new genotypes are selected by these new conditions or if they favor the expansion of existing genotypes. Also higher temperatures increase sandfly metabolism and reduce generation times that can result in larger vector populations besides possibly effecting positively or negatively parasite development. The recent expansion of populations of Lutzomyia longipalpis into urban areas in Brazil and other Latin American countries has created a new transmission scenario, but there is evidence that more than one genotype is involved. There are contradictory studies for risk factors associated with vectors of the same species. The risk factor for kalazar in South Asia may be either greater or smaller with the presence of large animals, such as cows or buffalos. It is not clear why this is but it may be because Phlebotomus agrentipes populations are modulated in different ways by the presence of large animals. Both these species are predominantly domestic and population sizes may also be related to the quantity of organic material that serves as food for their larvae. Many more parasites and vectors are linked to cutaneous leishmaniasis and there are a correspondingly greater numbers of novel transmission scenarios. L.tropica is typically a parasite of cities transmitted from man to man by P.sergenti. However, L.killicki, considered to be a L.tropica variant was found in silvatic areas where the vector is P.sergenti but this fly did not occur in the urban areas of the same vicinity. In the Americas we also see adaptation of silvatic N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina flies, such as Nyssomyia whitmani and N.intermedia, to peri-domestic sites favoring peri-domestic transmission but the sizes of silvatic and peri-domestic populations is not uniform throughout their geographical distribution. Potentially molecular markers could help us understand sandfly habitat and host preferences that are keys in understanding transmission cycles. ID 43-O Ecology & Epidemiology ID 42-O Ecology & Epidemiology 43 STUDIES ON THE SAND FLY FAUNA (DIPTERA: PSYCHODIDAE) IN HIGH-TRANSMISSION AREAS OF VISCERAL LEISHMANIASIS IN NORTH INDIA, SPECIAL EMPHASIS ON UTTARANCHAL REGION NS Singh1; Doris Phillips-Singh2; Ipe M Ipe3; Shubham Pandey4; Dinesh Lal5 1,2,4, EMERGING CUTANEOUS LEISHMANIASIS IN ISRAEL: A REVIEW OF THE LAST TWO DECADES Sandfly Research Laboratory, Lucknow Christian College Lucknow Christian College,, Lucknow - INDIA ; 3,5.School of Entomology, St. John's College, Agra-282002 – INDIA [email protected] Alon Warburg Department of Microbiology and Molecular Genetics, The Institute of Medical Research Israel-Canada, The Kuvin Center for the study of Infectious and Tropical Diseases, The Hebrew University of Jerusalem, Jerusalem, Israel [email protected] Keywords: Cutaneous Leishmaniasis, Leishmania major, Leishmaia tropica In Israel cutaneous leishmaniasis (CL) is caused by Leishmania major and L. tropica. Until the 1980s the vast majority of CL cases were caused by L. major and were restricted geographically to the Negev and Arava Deserts in Southern Israel. However, during the last three decades CL has expanded its range northwards in hitherto non endemic areas. The emergent foci of L. tropica in the Galilee region of Northern Israel, the central Palestinian West Bank and the Judean Desert affect both urban and rural environments, with most CL cases residing in peripheral houses bordering open areas. This distribution suggests that the infected sand flies are arriving from outside the inhabited area, having acquired the parasites by feeding on infected rock hyraxes (Procavia capensis), large populations of which are encroaching upon human habitations in many parts of the country. In an emerging focus of CL caused by L. major in the Beit She'an Valley of northern Israel, two rodent species, social voles (Microtus guentheri) and Tristram's jirds (Meriones tristrami), were incriminated as new reservoir hosts. The adaptation of L. major to these hosts has facilitated the emergence of new CL foci where the main reservoir hosts of the parasite (Psammomys obesus and Meriones crassus) are absent. In this talk I shall review studies documenting the emergence of these new foci. Keywords: Phlebotominae, Leishmaniasis, Uttrakhand, India Visceral Leishmaniasis in the Indian sub-continent, also known as Kala-azar, is caused by Leishmania donovani transmitted by Phlebotomus argentipes. This study endeavors to delineate the suitable locations of Phlebotomus argentipes with relation to environmental characteristics between endemic and non-endemic districts in India. The aim of this study was to evaluate the urbanization of VL vectors in North India, special emphasis on Uttaranchal Region where VL is endemic. Sandflies were captured using, light traps along an urban-rural gradient. The traps were installed outside and inside mud houses at a height of 200 cm. A total of 2709 sandflies were collected and 9 species were identified. The predominant species were Sergentomyia punjabaensis (rate of 67.12%) and P. argentipes (0.5%). A total of 1357 females were dissected and no Leishmania infections were detected. Most of the sand flies were captured at the border of a forest (76.37%) and no flies were captured in the urban area, which suggests that transmission is still restricted to rural sites. However, the fact that a specimen was collected in an intermediate area indicates that urbanization is a real possibility and that vector monitoring is important. The present study should be utilized to make critical decisions for vector surveillance and controlling Kala-azar disease vectors. At present in India, Bihar (31 out of 38 districts), West Bengal (10 out of 24 districts), Uttar Pradesh (4 districts), that border Bihar, and a small portion of the Jharkhand district are endemic at varying levels. This study provides the report evaluating the prevalence of Leishmania infection in sand flies in a region endemic for VL Uttaranchal region in India. Sergentomyia species are the most common species of sand fly. Knowledge of the infection rate in female P. argentipes may help in predicting severity of disease and in vector elimination programs. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 44-O Ecology & Epidemiology ID 45-O Ecology & Epidemiology INFECTION OF WILD SANDFLIES COLLECTED FROM DIFFERENT BIO-GEOGRAPHICAL AREAS OF TUNISIA WITH PHLEBOVIRUSES INVESTIGATIONS ON SAND FLY BIONOMICS AND LEISHMANIA NATURAL INFECTIONS IN EASTERN SICILY, ITALY, WITH PARTICULAR REFERENCE TO PHLEBOTOMUS SERGENTI W Fares1, RN Charrel2, K Dachraoui1, L Bichaud2, W Barhoumi1, M Derbali1, S Cherni1, I Chelbi1, X de Lamballerie2, E Zhioua1 1 Institut Pasteur de Tunis, Laboratory of Vector Ecology, 13 Place Pasteur BP 74, 1002 Tunis, Tunisia. 2Aix Marseille University, IRD French Institute of Research for Development, EHESP French School of Public Health, EPV UMR_D 190 [email protected] 44 Bongiorno G1, Lisi O2, Severini F1, Vaccalluzzo V2, Khoury C1, Di Muccio T1, Gradoni L1, Maroli M1, D'Urso V2, Gramiccia M1 1 Unit of Vector-borne Diseases and International Health, Istituto Superiore di Sanità, Rome; 2Department of Biological, Geological and Environmental Sciences, Section of Animal Biology “M. La Greca”, University of Catania; Italy [email protected] Keywords: Sandflies, phlebovirus, emerging, Mediterranean basin. An entomological investigation performed in 2013 covering different bio-geographical areas varying from humid in the north to the arid in the center showed that sandflies of the subgenus Larroussius including Phlebotomus perniciosus, Phlebotomus perfiliewi, and Phlebotomus longicuspis are abundant and widely distributed in Tunisia. A total of 3,992 collected and pooled with up to 30 specimens per pool based on sex, trapping location and collection date were tested for the presence of phleboviruses by nested reverse transcriptase polymerase chain reaction and sequencing. Of a total of 135 pools, 23 were positive, yielding a minimum infection rate of 0.6%. Phylogenetic analysis performed using partial amino acid sequence in the polymerase gene showed that all these phleboviruses were grouped in one cluster clearly distinct but closely related to Massilia virus and Granada virus. This putative novel virus, tentatively called Saddaguia virus (SADV), is widely distributed in Tunisia. Together with Toscana, Punique, and Utique viruses, SADV is the fourth recognized phlebovirus to be transmitted by sandflies in Tunisia. The medical and public health interest of SADV remains to be investigated. Keywords: Eastern Sicily, Cutaneous Leishmaniasis, Phlebotomus perniciosus, Phlebotomus sergenti Visceral and cutaneous leishmaniasis (CL) by Leishmania infantum have long been known to be endemic in Sicily. Catania province, sited on the eastern coast of the island, represents also an old endemic territory for Phlebotomus sergenti, the sand fly vector of CL by L.tropica. Because of the increasing human migration to Sicily and the related risk for the introduction of exotic Leishmania, an investigation on sand fly fauna composition, seasonal dynamics and Leishmania infections was addressed in CL endemic territories. Four urban (Catania city) and 11 periurban/rural sites, located in a previously recorded P.sergenti area, were investigated. Sand flies were collected by sticky traps from April through December and by CDC light traps from June through October 2013. Live females were dissected and examined for Leishmania promastigotes identified by ITS1 PCR- RFLP, whereas dead females were analyzed by Leishmania nested (n)-PCR.Out of 4105 specimens collected, 2545 (62.0%) were Sergentomyia minuta. Among 1560 Phlebotomus specimens, P.perniciosus (65.4%) and P.sergenti (20.8%) were the most prevalent followed by P.papatasi, P.neglectus and P.mascittii. P.sergenti was confirmed to be endemic, being recorded in 3 urban and 5 periurban/rural sites of which those located at the slopes of mount Etna were the most productive. The comparative dynamics of P.sergenti and P.perniciosus showed that they peaked in different months, July and August respectively. Females captured alive from P.sergenti-positive sites (143 P.sergenti and 34 P.perniciosus) were found negative at dissection. In a P.sergenti-negative site dominated by P.perniciosus 3/118 (2.5%) specimens of this species were detected with promastigotes identified as L. infantum. Out of 175 dead females from all sites, 17 (9.7%) were found positive by Leishmania n-PCR, of which 13 were P.perniciosus and 4 P. sergenti. While the vector role of P.perniciosus in the L.infantum transmission was confirmed, that of P.sergenti remains unclear. The detection of Leishmania DNA in this species suggests an implication in parasite circulation, however P.sergenti does not seem to play a role as an active vector of CL, which has been recently confirmed to be caused by L.infantum in the Catania area. This study was carried out in the frame of EDENext, a collaborative project of the 7th FP (2011-2014) funded by the European Commission under the DG Health; Contract Number: 261504. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 45 ID 46-O Vector Competence & Infections ID 47-O Vector Competence & Infections N AT U R A L I N F E C T I O N O F L U T Z O M Y I A C AY E N N E N S I S C AY E N N E N S I S ( D I P T E R A : PSYCHODIDAE) WITH LEISHMANIA (VIANNIA) SPP., POTENTIAL VECTOR IN THE ENZOOTIC CYCLE OF LEISHMANIASIS IN URBAN AND RURAL HABITATS OF THE COLOMBIAN CARIBBEAN PRIORITIES FOR PROGRESSING FROM ASSESSMENTS OF VECTOR COMPETENCE TO THE TRANSMISSION MODELLING AND CONTROL OF LEISHMANIASIS Paul D Ready Alveiro Pérez-Doria 1, Margaret Paternina 1, Lily Martinez 1, Luis Paternina1, Daniel Verbel-Vergara1, Suljey Cochero2, Eduar E. Bejarano1 1 Investigador, Grupo de Investigaciones Biomédicas, Universidad de Sucre, Sincelejo, Colombia. 2Entomóloga, Departamento Administrativo de Seguridad Social en Salud de Sucre – DASSSALUD, Sincelejo, Sucre, Colombia. [email protected] Keywords: Natural infection, Leishmania (Viannia), Lutzomyia cayennensis cayennensis, cutaneous leishmaniasis, urban foci. The first recorded observations in the literature on bloodmeal sources of Lutzomyia spp. indicate that the members of the subgenus Micropygomyia Barretto, 1962 feed on reptiles. Since then, and despite the recent finding of natural infections of Lu. cayennensis cayennensis with trypanosomatid parasites, the vector potential of this species has been underestimated due to the low number of insects captured with CDC light traps in entomological surveys. By contrast, this was the most abundant species in samples from diurnal resting sites collected in an autochthonous urban and a rural focus of cutaneous leishmaniasis in the municipalities of Ovejas and Los Palmitos respectively, placed in the Montes de María region of the Sucre, Colombian. This facilitated the search for natural infection with Leishmania. In all 274 females of the genus Lutzomyia were captured, dissected and identified in search of trypanosomatid parasites. In Ovejas 160 females were collected, Lu. micropyga was the most abundant (42.6%) followed by Lu. evansi (22.8%), Lu. c. cayennensis (22.2%), Lu. trinidadensis (11.7%) and Lu. panamensis (0.6%). In this locality natural trypanosomatid infection rates of 4/69 (5.8%) and 2/36 (5.6%) were found in Lu. micropyga and Lu. c. cayennensis, respectively. In Los Palmitos, the 114 females captured included Lu. evansi (91.0%), Lu. trinidadensis (0.9%) and Lu. c. cayennensis (8.0%), the last of these species, with a natural trypanosomatid infection rate of 2/9 (22.2%). In order to identify the parasites a 350 bp fragment of Internal Transcript Spacer 1 (ITS1) was amplified and sequenced. The parasites found in two females of Lu. c. cayennensis were identified as belonging to the subgenus Leishmania (Viannia), as suggested by their peripyloric distribution in the sand fly gut. Consequently in Ovejas Lu. c. cayennensis presented a Le. (Viannia) spp. infection rate of 2.8%, while in Los Palmitos it was 11.1%. In conclusion, Lu. c. cayennensis can be naturally infected with parasites of the subgenus Leishmania (Viannia) and can support their development, satisfying two of the most important criteria in the incrimination of a vector. Given the finding of natural infections in females captured on diurnal resting sites, we suggested that Lu. c. cayennensis probably acts as a vector in the maintenance of the urban and rural zoonotic cycles of cutaneous leishmaniasis in the study area and should be taken into account in the control programs for leishmaniasis. Disease Control Department, Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London WC1E 7HT, United Kingdom [email protected] Keywords: vector competence, transmission modelling, risk factors, vector control There has been much elegant research on the vectorial competence of sand flies, which has been aimed at elucidating the factors that permit Leishmania infections firstly to become established in the abdominal midgut or hindgut of female flies, and secondly to develop infective parasite forms and promastigote secretory gel in the thoracic midgut in order to permit infections to establish in mammalian hosts. There has also been fascinating research on how female sand flies locate hosts and physically transmit Leishmania, a process that can be manipulated by the parasite. In contrast, there has been little research on modelling the transmission of Leishmania, in order to distinguish between those sand fly populations that are epidemiologically important and those that only have the potential to be so. Only the former need be targeted for control measures. This distinction is illustrated by Lutzomyia longipalpis: Laboratory populations are competent vectors of various Leishmania species of the subgenera Leishmania and Viannia, but wild populations are only usually associated with the transmission of Leishmania infantum chagasi. There is a need to prioritise basic research in order to identify sand fly populations to be targeted for control, and this talk offers suggestions. Much can be learned from malaria control, including the realization that the aim of sand fly control should not be to reduce the relative abundance of a vector species, but to reduce the contact of its blood feeding females with humans and to reduce the lifespans of the female flies so that fewer survive long enough to transmit infective parasites. References: Ready PD. Biology of phlebotomine sand flies as vectors of disease agents. Annu Rev Entomol. 2013;58:227-250.Ready PD. Epidemiology of visceral leishmaniasis. Clin Epidemiol. 2014;6:147154.Ready PD, Rogers ME. 2012. Behaviour of sandflies infected with Leishmania. In Ecology and Control of Vector-Borne Diseases. Vol. 3: Ecology of Vector-Parasite Interactions, ed. CJM Koenraadt, W Takken. Wageningen, The Neth.: Wageningen Acad. Publ. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 46 ID 48-O Leishmania – Sandfly Interactions ID 49-O Vector competence & Infections LEISHMANIA CHAGASI IN ITS NATURAL VECTOR LUTZOMYIA LONGIPALPIS. XENODIAGNOSIS OF LEISHMANIA DONOVANI INFECTIONS IN ASYMPTOMATIC MICE USING PHLEBOTOMUS ORIENTALIS SAND FLIES Paulo F P Pimenta, Vanessa C Freitas, Nágila F C Secundino Laboratory of Medical Entomology, Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz – FIOCRUZ-MG. Av. Augusto de Lima, 1715 CEP 30190-002 Belo Horizonte, M.G. Brazil [email protected] Keywords: Leishmania chagasi, sandfly, interaction, Lutzomyia longipalpis Studies on Leishmania-sandfly vector interactions are important to understand processes involved in parasite development and transmission. Considerable information has been reported mainly for Old World Leishmania species. In the New World, there is little detailed information about the development of Leishmania chagasi in its natural vector Lutzomyia longipalpis. The development of this parasite was investigated in experimental infection of the vector with cultured promastigotes and axenic amastigotes obtained by in vitro transformation. Both parasite forms enable high percentage of infected sand flies. It was observed a decrease in parasite number within the gut on the third day after bloodmeal. However, the surviving promatigotes of both groups were able to multiply and develop in the gut. The highest density of parasite per gut occurred between six and seven days and all the promastigote forms were observed in the parasite life cycle. Experimental infections initiated with promastigotes and axenic amastigotes showed similar trends of development and produced infective forms. Therefore, promastigote infection can be preferably used in experimental infection being less laboring. For many Leishmania species, the lipophosphoglycan (LPG) has been suggested to promote the attachment between parasite and sand fly epithelial midgut. Structural analyses of different Leishmania species LPG have showed that the polymorphism in theses glycoconjugates reside in the sugar side chain and cap. The analysis of strain BH46 side chains demonstrated for the first time a poly glucosylated LPG (1, 2 and 3 glucose side chains) in L. chagasi. The LPG importance in survive of this specie in L. longipalpis was investigated using mutant parasite in LPG biosynthesis. It was observed reductions in mutant survive or growth prior to excretion and, in 60 hour, the infection was completely lost. In all natural Leishmania/vector combinations examined to date, the LPG is required to binding the parasites to the sand fly midgut to avoid their loss with the blood meal excretion. Unlike of these observations, we observed that the LPG synthesis was essential to early survive of the L. chagasi in L. longipalpis midgut, probably protect the parasites from enzymatic attack. Morphological aspects of Leishmania/vector interaction were investigated using light microscopy and scanning and transmission electron microscopies. Differently of highly specialized attachment proposed previously to L. chagasi in L. longipalpis, few parasites were observed with body and flagella in contact with the epithelial midgut and, not often, with the flagella superficially inserted between microvilli. In front of no effective vaccine against the disease and a limited range of drugs to treatment, details in all aspects of parasite biology are important to formulate new strategies of control against protozoa and vector. Jovana Sadlova1, Veronika Hrobarikova1, Jan Votypka1, Alon Warburg2 and Petr Volf1 1 Department of Parasitology, Faculty of Science, Charles University in Prague, 128 44 Prague 2, Czech Republic. 2Department of Microbiology & Molecular Genetics, The Institute for Medical Research Israel-Canada, The Kuvin Centre for the Study of Infectious & Tropical Diseases, The Hebrew University- Hadassah Medical School, The Hebrew University of Jerusalem, 91120, Israel. [email protected] Keywords: Xenodiagnosis, Visceral leishmaniasis, Asymptomatic hosts, Reservoir hosts In endemic areas of visceral leishmaniasis most infected hosts remain asymptomatic but potentially infectious to biting sand flies. Therefore, their role in the epidemiology of the disease warrants intensive study. The best way to test infectiousness of asymptomatic hosts for sand flies is by xenodiagnosis. However, in humans this test is problematic for ethical and logistic reasons. In the present study we established a model using BALB/c mice experimentally infected with Leishmania donovani to study their infectiousness for Phlebotomus orientalis, the vector of visceral leishmaniasis in north Ethiopia and Sudan. To mimic the natural conditions of host infection, mice were inoculated with parasites isolated from the thoracic midguts of experimentally infected sand flies with predominance of metacyclic forms. Although mice did not show any signs of disease, around 20% of the sand fly females feeding at the site of infection acquired L. donovani infections. The majority of L. donovani-positive P. orientalis females (76 %) had heavy infections with the stomodeal valve fully colonized with attached parasites. Inoculated ears remained infective for sand flies till the end of the experiment at week 15 post infection. By week 12 post infection females feeding on the contralateral ear of two mice were also infected, but females feeding on other parts of the body remained negative. In sacrificed mice the highest parasite numbers were found in inoculated ears and draining lymph nodes; infections were detected also in the spleen, liver and blood, but scarcely in the contralateral ear; and, the other organs tested were negative. The study showed that asymptomatic Balb/c mice harbour parasite numbers sufficient to infect P. orientalis. This finding also encourages searching for more data about the role of rodents in L. donovani circulation. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 50-O Vector competence & Infections ID 51-O Vector competence & Infection DEVELOPMENT OF LEISHMANIA PARASITES IN BITING MIDGES, CULICOIDES NUBECULOSUS AND CULICOIDES SONORENSIS (DIPTERA: CERATOPOGONIDAE) AND IMPLICATIONS FOR SCREENING VECTOR COMPETENCE THE SAND FLY LUTZOMYIA EVANSI (DIPTERA: PSYCHODIDAE), AS A PERMISSIVE VECTOR OF LEISHMANIA INFANTUM AND LE. BRAZILIENSIS IN BOLÍVAR, V Seblova-Hrobarikova, J Sadlova, S Carpenter, P. Bates and P Volf Luis Roberto Romero1, Jorge Luis Rodríguez1, Alveiro Pérez-Doria1, José G. Vergara, Ana Montesino1, Lily Paola Martínez1, Luis Cortés2, Suljey Cochero 1,3 & Eduar Elías Bejarano1 Department of Parasitology, Charles University in Prague, Faculty of Science, Prague 2, 128 44, Czech Republic [email protected] 47 COLOMBIA 1 Grupo de Investigaciones Biomédicas, Universidad de Sucre, Sincelejo, Colombia; Secretaría de Salud Departamental de Bolívar, Cartagena, Colombia; 3Secretaría de Salud Departamental de Sucre, Sincelejo, Colombia. 2 Keywords: biting midges, Leishmania parasites, vector competence Biting midges of the genus Forcipomyia (Diptera: Ceratopogonidae) have recently been implicated as vectors of kinetoplastid parasites in the Leishmania enriettii complex. Here, we assessed susceptibility of two colonized midges, Culicoides nubeculosus and C. sonorensis, to Leishmania infection. Midges C. nubeculosus and C. sonorensis were experimentally infected with two strains from the L. enriettii complex, L. australiensis originally from red kangaroos (LV756) and L. enriettii from guinea pigs (LV90), and two Leishmania species infecting humans, L. major (FVI) and L. infantum (CUK3). A permissive species of sand fly, Lutzomyia longipalpis, was used as a control. Engorged females were microscopically examined at different time points post-bloodmeal to assess the localization of Leishmania promastigotes in the gut. In C. nubeculosus, all four tested Leishmania strains produced only light infection in the abdominal midgut with presence of long nectomonads. In contrast, C. sonorensis was able to support late-stage development of both strains from the L. enriettii complex; in 20% of females, parasites migrated anteriorly and colonized the stomodeal valve, which is one prerequistite for subsequent transmission. Similarly, L. longipalpis was susceptible to parasites from the L. enriettii complex but only to strain LV756. In summary, we showed that the palaearctic species of biting midge C. nubeculosus is not susceptible to any Leishmania species tested. In contrast, L. longipalpis and C. sonorensis supported late stage lifecycle development of parasites from the L. enriettii complex, which is promising for their transmission potential. However, detailed transmission experiments are needed to avoid mistaken speculations about vector competence of these species for members of the Leishmania enriettii complex. [email protected] Keywords: Lutzomyia evansi, permissive vector, Montes de María. Los Montes de María on the Caribbean coast of Colombia is considered to be a mixed focus of leishmaniasis transmission due to the presence of cases of both the cutaneous and visceral forms of the disease. In this area of the country Leishmania infantum, the aetiological agent of visceral leishmaniasis, is known to be transmitted by Lutzomyia evansi. However little is known about the vectors of Le. braziliensis and Le. guyanensis, aetiological agents of cutaneous leishmaniasis, the clinical form with the greatest number of human cases. This study focused on the detection and identification of trypanosomatids in phlebotomines from a municipality of the Montes de María region. Four collections were made with Shannon trap and by sampling from diurnal resting sites with an electronic aspirator in the Las Margaritas neighbourhood of El Carmen de Bolívar (9°43'58,808''N; 75°07'32,962''W), during two rainy and two dry seasons in 2012-2013. The guts of female phlebotomines were extracted and examined under the microscope for trypanosomatid parasites, as well as the spermathecae for taxonomic determination of the species. The parasites found in the gut were genotyped using PCR and sequencing in tandem of a region of the cytochrome b gene and another of subtelomeric repetitions. In all 2509 sand fly specimens were collected, of which 88.9% were Lu. evansi, 2.0% Lu. cayennensis cayennensis, 0.7% Lu. gomezi and 0.2% Lu. panamensis. Two specimens of Lu. evansi were found naturally infected with trypanosomatids. One presented a suprapyloric distribution of the parasites, identified by molecular techniques as Le. infantum, while in the other specimen the trypanosomatids were distributed peripylorically and found to be Le. braziliensis. These findings demonstrate the permissive character of Lu. evansi as a vector of two Leishmania species and its importance in the epidemiological cycle of cutaneous leishmaniasis in the Colombian Caribbean. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 48 ID 52-O Vector competence & Infection ID 53-O Vector-Parasite-Host Interactions EXPERIMENTAL SUSCEPTIBILITY OF LUTZOMYIA (L.) LONGIPALPIS TO DIFFERENT SPECIES OF LEISHMANIA THE PROSPECT FOR A SAND FLY SALIVA-BASED VACCINE AGAINST LEISHMANIASIS IN HUMANS Ana Clara Araújo Machado Pires, Paulo Filemon Paolucci Pimenta and Nágila Francinete Costa Secundino Shaden Kamhawi, Fabiano Oliveira and Jesus Valenzuela Laboratory of Medical Entomology of the Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz – MG Vector Molecular Biology Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health.NIAID, NIH- [email protected] Keywords: Susceptibility, Lutzomyia (L.) longipalpis, Leishmania In order to better understand the epidemiology of leishmaniasis and the parasite transmission it is important to develop studies on the Leishmania-vector interaction. The knowledge of the process of the sand fly-parasite interaction can contribute for new control strategies of leishmaniasis, a complex disease that until now has no effective vaccine and limited range of drugs for treatment. In nature, some sandfly species show remarkable specificity to transmit an exclusive Leishmania species (non-permissive vector), while others can be vectors of more than one parasite species (permissive vector). However, in the laboratory, some sandfly like Lutzomyia (L.) longipalpis, exclusive vector of L. (L.) chagasi, can be infected experimentally by different parasite species. One important step for Leishmania to establish infection in sandfly vector is to be capable to attach to the midgut epithelium, which is known to be dependent of LPG molecules in non-permissive vectors. However, the parasite attachment in the permissive vectors appears to be different and a nonLPG dependent mechanism. Here, we describe details of the development of four distinct species of Leishmania: Leishmania (L) major, L. (L.) amazonensis, L. (Viannia) braziliensis and L. (L.) chagasi infantum in the sandfly Lu. (L.) longipalpis, a lab-permissive vector. Experimental infections using membrane feeding device were conducted using three parasite doses (4x107, 2x107 and 1x107) mixed with mouse blood. On day 6 after the complete digestion of the infective blood meals, it was observed that L. (L.) chagasi infantum, L. (L.) major and L. (L.) amazonensis, in all tested parasite doses, infected the Lu. (L.) longipalpis. However, in L. (V.) braziliensis only the highest dose (4x107) was able to develop infection in the vector but with a low average of infection rate (1,000 parasites/sandfly). In contrast, even with the lowest parasite dose, L. (L.) amazonensis and L. (L.) major showed an average of 7,000 parasites/sandfly, while L. (L.) chagasi showed an average of 10,000 parasites/sandfly. Although it is well known that Lu (L.) longipalpis in the nature can only transmit L. (L.) chagasi it is still need further studies to prove that this vector can transmit other Leishmania species. Financial SUPPORT: FAPEMIG, FIOCRUZ, CNPq and Capes. [email protected] Keywords: Vector saliva, DTH response, Th1 immune response, Leishmania vaccine To date, a human vaccine against any form of leishmaniasis is not available. Immunity to saliva or distinct salivary proteins has a profound impact on host immunity to the Leishmania parasites. In most cases, saliva-specific immunity drives a protective immune response against Leishmania. Protective molecules have mostly shared a similar property, the induction of a delayed-type hypersensitivity (DTH) response biased towards a Th1 profile. This protection has been observed in rodents and non-human primates and accelerates the development of a robust Leishmania-specific immune response with little pathology. Importantly, immunity to a TH1-DTHinducing salivary protein has protected against both cutaneous and visceral leishmaniases suggesting that it is acts on parasites in the skin soon after an infected bite when they are at their most vulnerable. The strong protection mediated by immunity to saliva in various models argues for their inclusion in a leishmaniasis vaccine. The opportunities and challenges facing a combination salivaLeishmania vaccine will be discussed. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 54-O Vector-Parasite-Host Interactions ID 55-O Vector-Parasite-Host Interactions CHARACTERIZATION OF LUTZOMYIA LONGIPALPIS IMMUNE RESPONSES WITH EMPHASIS IN LEISHMANIA INFECTION P R O M A S T I G O T E S E C R E TO RY G E L F R O M INFECTED SAND FLIES SUBVERTS THE WOUND HEALING RESPONSE IN AN INSULIN GROWTH FACTOR1-DEPENDENT MANNER Pitaluga AN, Tinoco B, Di Blasi T, Silva AM, Tempone A, Traub-Csekö YM Instituto Oswaldo Cruz, Fiocruz [email protected] 49 Matthew E Rogers, Emilie Giraud, Rod J Dillon, Paul A Bates, Tamsyn Derrick, Oihane Martin, Ingrid Műller London School of Hygiene and Tropical Medicine [email protected] Keywords: Lutzomyia longipalpis, innate immunity Keywords: Leishmania transmission, PSG, IGF1 Lutzomyia longipalpis is the major vector for visceral leishmaniasis in Brazil. We are studying immune responses both in the insect vector and in the L. longipalpis LL5 embryonic cell line. Insect innate immune response pathways are Toll, IMD and JAK-STAT. We have identified an IMD and Toll response in LL5 cells through the silencing of the respective repressor genes caspar and cactus. This silencing produced the expected effect of increasing the expression of the antimicrobial peptides (AMPs) defensin and cecropin, thus establishing LL5 cells as a valid model to study sandfly immunity. We also investigated de effect of silencing these repressors in female adult insects. We have previously determined a role for the IMD pathway in the vector infection by Leishmania through the silencing of the repressor caspar, which decreased infection. We now silenced cactus and, surprisingly, as opposed to LL5 cells, this led to the decrease of AMPs production. As a possible explanation, we verified that WntD, an inhibitor of the Toll pathway identified in Drosophila, had increased expression upon cactus silencing. This might be related to the preservation of the insect microbiota, which was actually decreased when cactus was silenced. We have also studied the effects of Leishmania infection on L. longipalpis. Infection caused an early increase of cactus expression followed by a return to normal levels that was accompanied by a higher expression of AMPs. Interestingly the infection by Leishmania also caused an early growth of the microbiota, probably related to the increase of cactus expression. We were intrigued by the apparent capacity of Leishmania to increase the expression of cactus and thus inhibit the Toll pathway. It is known that Leishmania Gp63 activates a macrophage tyrosine phosphatase (SHP-1), capable of inhibiting the Toll and Jak/STAT pathways. L. longipalpis has a homoloque of SHP-1, which has increased expression during L. longipalpis infection by L. i. chagasi, indicating the possible conservation of a mechanism of Leishmania infection control in mammals and insects. In the sand fly midgut secreted proteophosphoglycans from Leishmania parasites form a biological plug known as the promastigote secretory gel (PSG), which facilitates the regurgitative mode of transmission and infection of the mammalian host. Although the disease-exacerbating properties of PSG have been related to macrophage alternative activation the underlying signalling pathways remain unknown. Affymetrix Mouse GeneChip and Pathway analysis revealed differential expression of 7,927 transcripts (fold change >1.5, p<0.05) to Leishmania mexicana PSG in BALB/c mouse ears (27% of the mouse genome). These transcripts were transiently expressed during the first 6 hours and enriched for pathways involved in inflammation, cell proliferation, fibrosis, epithelial cell differentiation and wound remodelling. We found that L. mexicana PSG significantly accelerated wound healing in vitro and in vivo; which was associated with an early up-regulation of transcripts involved in inflammation (IL-1a, IL-6, TNFa) and inflammatory cell recruitment (CCL2, CCL3, CCL4, CXCL2), followed 6 days later by enhanced expression of transcripts associated with epithelial cell proliferation, fibroplasia and fibrosis (FGFR2, EGF, EGFR, IGF1). Antibody blockade of IGF1 ablated the gel's ability to promote wound closure in BALB/c mouse ears and significantly reduced the virulence of L. mexicana infection delivered by an individual infected sand fly bite. Dermal macrophages recruited to air pouches on the backs of mice revealed that IGF1 was pivotal to the PSG's ability to promote macrophage alterative activation and L. mexicana infection. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 56-O Vector-Parasite-Host Interactions ID 57-O Vector-Parasite-Host Interactions V E C TO R - T R A N S M I S S I O N O F L E I S H M A N I A DONOVANI TRIGGERS A DISTINCT IMMUNE RESPONSE THAT FAVORS PARASITE SURVIVAL AND DISSEMINATION CYTOKINE LEVELS IN RESPONSE TO SALIVARY GLAND EXTRACTS AND TO LEISHMANIA MAJOR PARASITES IN INDIVIDUALS LIVING IN ENDEMIC A R E A S F O R Z O O N O T I C C U TA N E O U S LEISHMANIASIS IN TUNISIA Ranadhir Dey1, Hamide Aslan2, Amritanshu Joshi, Claudio Meneses2, Pradeep K. Dagur3, John Philip McCoy3, Robert Duncan1, Jesus G. Valenzuela2, Hira L. Nakhasi1* and Shaden Kamhawi2* 1 Vector Molecular Biology Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health.NIAID, NIH. 2Vector Biology, National Institute of Allergy and Infectious Diseases, Baltimore, USA; [email protected] 50 Wafa Rebai-Kammoun, Ikbel Naouar, Darragh Duffy, Valentina Libri, Milena Hasan, Amine Toumi, Afif B. Salah, Matthew Albert, Hechmi Louzir, Amel Meddeb-Garnaoui Laboratory of Medical Parasitology, Biotechnology and Biomolecules, Institut Pasteur de Tunis; Tunisia;Laboratory of Transmission Control and Immunobiology of Infection, Institut Pasteur de Tunis; Tunisia; Laboratory of Dendritic Cell Biology, Department of Immunology, Institut Pasteur, Paris, France; : Center for Human Immunology, Institut Pasteur, Paris, France [email protected] Keywords: Vector-tranmission, Leishmania donovani, immune response, visceral leishmaniasis Keywords: Phlebotomus papatasi, Salivary gland extract, Leishmania, immune response Visceral leishmaniasis (VL) caused by Leishmania donovani is prevalent in the Indian subcontinent, South Asia, and East Africa and is considered mostly an anthroponosis. Most VL models of infection involve intravenous, intracardiac, or intraperitoneal injection of 106–108 parasites into laboratory animals. These routes bypass the skin, the natural delivery site of the parasites by sand fly bites. For cutaneous leishmaniasis, natural transmission by vector bites revealed important distinctions in disease initiation and progression and abrogated protection observed in vaccinated animals following needle challenge. Here, we infected BALB/c mice via bites of L. donovaniinfected Lutzomyia longipalpis sand flies and compared disease development to intravenous (i.v.) or intradermal (i.d.) needle-initiated infections. The mice infected via sand fly bites- developed Leishmania-specific antibodies that correlated with dissemination of parasites into the spleen and liver; by weeks 20-30 post-infection, most mice cleared liver parasites but displayed a slight increase in the splenic parasite burden. Following i.v.-injection, the parasites persisted in both the spleen and liver at 20-30 weeks post-challenge while i.d.-injected parasites failed to disseminate to visceral organs. The immune response to vector-initiated infections was silent with splenic CD4+ and CD8+ T cells producing less IFN-g and TNF-a at 5 and 30 weeks post-infection but similar levels of IL-10 compared to the i.v.-initiated infection. In the skin, vector-initiated infection induced an acute pro-inflammatory response (IFN-g, IL1-b, TNF-a and NOS2) 3h post-transmission that was not observed following i.d.injection of parasites. This response subsided at 6h to be replaced by a persistent recruitment of neutrophils and alternately activated monocytes to the bite site characterized by a sustained induction of IL10, MCP1, Ym1 and ILRN1 up to 18h post-infection. Altogether, vector-transmission induces an acute pro-inflammatory response followed by a persistent and immunologically quiet cellular infiltration at the bite site that is distinct from the immune response to i.v.- and i.d.-initiated infections. This immunologically silent environment may facilitate the dissemination and survival of L. donovani in its mammalian hosts. We analyzed cellular proliferative and IFN-γ and IL-10 responses (ELISA) after stimulation by salivary gland extracts (SGE) from Phlebotomus papatasi, vector of L. major, in 378 individuals living in endemic areas for cutaneous leishmaniasis (CL) due to L. major. These were subdivided according to the leishmanin skin test (LST) response and the presence of scars: healed (HCL: LST+/Scars+), asymptomatics (As: LST+/Scars-) and naïves (LST-/Scars-). A significant IFN-γ response to SGE was only observed in naive individuals. IL10 was not detected in response to SGE in all analyzed groups. To further analyze cytokine response to SGE, we used Luminex technology to measure IL-1b, IL-2, IL-5, IL-6, GM-CSF, IL-13, TNF-a, IL-12, IL-18, IFN-γ and IL-10 levels in supernatants from SGE-stimulated PBMC derived from HCL (n=17), As (n=23) and Naives (n=18). HCL and As individuals were further selected on the basis of a strong positive IFN-γ responses to L. major parasites. Our results showed that all cytokines analyzed were not detected after SGE stimulation in these groups. The significant IFN-γ response detected by ELISA could be explained by the larger number of individuals tested. Interestingly, a clinical follow up of these individuals allowing to detect new CL case have shown that no new CL cases were observed among the naïve individuals, who showed positive anti-SGE proliferative responses unlike those with negative proliferative responses (6 new cases). These results suggest that cellular responses developed against salivary components in the absence of parasites may induce, under certain conditions, a protection against subsequent Leishmania infection. We also evaluated cytokine levels in supernatants from L. major-stimulated PBMC. A significant response was detected for IFN-γ, IL-1b, IL-2, IL-5, IL-10, GM-CSF, IL-13, TNF-a, IL-12 and IL-18 in HCL and As individuals. Interestingly, GM-CSF levels were significantly higher in HCL compared to As individuals. GM-CSF is an important hematopoietic growth factor and immune modulator. Some data support a protective effect of GM-CSF on the course of Leishmania infection. Analysis of these cytokine profiles may allow to identify biomarkers associated with disease outcome and to better understand the mechanisms of immune response against leishmaniasis. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 58-O Vector-Parasite-Host Interactions ID 59-O Vector-Parasite-Host Interactions P R O S P E C T I V E S T U D Y O F A C O H O RT O F INDIVIDUALS AT RISK OF ZOONOTIC CUTANEOUS LEISHMANIASIS IN TUNISIA TO EVALUATE CLINICAL AND IMMUNOLOGICAL PARAMETERS AND IDENTIFY CORRELATES OF PROTECTION. THE POSSIBLE ROLE OF A FLAGELLAR PROTEIN FROM LEISHMANIA SP. (FLAG) IN THE PARASITE SAND FLY INTERACTION Wafa Rebai Kammoun, Ikbel Naouar, Amel Meddeb-Garnaoui, Amine Toumi, Nabil Belhadj Hamida, Afif B Salah, Hechmi Louzir Laboratory of Medical Parasitology,Biotechnology and Biomolecules, Institut Pasteur de Tunis, Tunisia ;Laboratory of Transmission Control and Immunobiology of Infection, Institut Pasteur de Tunis, Tunisia 51 Tatiana Di-Blasi1; Antonio Jorge Tempone1; Erich Loza Telleria1; Fabricio K Marchini2; José Marcelo Ramalho-Ortigão3; Yara Maria Traub-Csekö1 1 Instituto Oswaldo Cruz - Fiocruz, Rio de Janeiro - RJ - Brazil; 2Instituto Carlos Chagas - Fiocruz, Curitiba - PR - Brazil; 3Kansas State University, Manhattan - KS United States of America. [email protected] [email protected] Keywords: Sand fly, Leishmania, Interaction, Flagellum Keywords: Zoonotic cutaneous leishmaniasis, endemic areas, Leishmanin Skin Test, correlates of protection The Leishmaniases constitutes a heterogeneous group of diseases that affect millions of people and are caused by various Leishmania (L.) species. Leishmania major is the causative agent of zoonotic cutaneous leishmaniasis (ZCL). The aim of this study was to establish and follow a prospective cohort of individuals at risk, living in endemic areas of L. major, to evaluate markers of exposure to Leishmania infection. In 790 individuals in 5 different endemic areas for L. major, we observed: positive Leishmanin Skin Test (LST):431 individuals and 190 had leishmaniasis typical scars. We observed an increase of GrB, IFNγ and IL10 production with the age of the study subjects (p=0.002, p=0.001 and p=0.019 respectively) and with the size of LST reactions for GrB and IFNγ (p<0.001). Moreover, a positive correlation between LST and both IFNγ and GrB production was observed. When considering the foci type, individuals living in Mnara, the oldest focus, responded with the highest levels of IFNγ, GrB but also IL10 which seems to be an indicator of exposure to Leishmania infection (p<0.001). We also observed a strong correlation between IFNγ and GrB (rho=0.80). After a transmission season, 29 ZCL cases were observed. IFNγ/IL10 and GrB/IL10 ratios were also calculated in order to investigate the potential correlation of these ratios with other with the resistance against ZCL. Interestingly, the majority of individuals who developed ZCL lesions had an IFNγ/IL10 ratios <10 (the lowest ratios), when only 7% of emerging ZCL cases had very high ratios. A significant decrease of ZCL cases is observed with the increase of IFNγ/IL10 ratio. Leishmaniasis diseases are caused by Leishmania protozoans and transmitted to the vertebrate host mainly through the bite of Lutzomyia (New World) or Phlebotomus (Old World) sand fly species. These parasites are ingested by the sand fly during blood feeding and adhere to the midgut epithelium through the flagellum which is an important step to avoid parasite excretion after blood digestion. Previous studies identified a Leishmania flagellar protein called FLAG potentially involved in the parasite adhesion to the sand fly gut. To verify the role of FLAG in the midgut adhesion process, we performed ex vivo and in vivo inhibition assays using an anti-FLAG monoclonal antibody. The experiments were performed with the pairs Leishmania infantum chagasi - Lutzomyia longipalpis, and Leishmania major - Phlebotomus papatasi, in which the parasites were previously incubated with the anti-FLAG antibody. The results obtained from both ex vivo and in vivo assays showed that only the pair L. major and P. papatasi presented a significant reduction in the number of parasites inside the gut when they were pre-incubated antiFLAG. In parallel, we performed pull down experiments in order to identify L. longipalpis and P. papatasi molecules that could interact with FLAG. For this purpose, we used extracts from L. longipalpis (LL5) and P. papatasi (PP9) cultured embryonic cells. In the experiments with LL5 extracts we observed only nonspecific bindings to the fusion protein or to the resin and no specific binding to FLAG. However, in the assays with PP9 extract we could identify possible FLAG binding molecules that were characterized through mass spectrometry. Among the molecules identified through the sequencing there are: PDGF- and VEGF-receptor related protein known as having a role in oogenesis and cell migration in Drosophila melanosgater. Another molecule identified was a fas-associated protein related with immunological response in Aedes aegypti. The interaction between parasite and its vector is a key step involved in the establishment of leishmania in the sand fly. Therefore the study of molecules involved in this interaction can contribute to develop new strategies to block parasite transmission. Financial support: CNPq; FIOCRUZ. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 52 ID 60-O Vector-Parasite-Host Interactions ID 61-O Genome & Genetics THE EFFECT OF TEMPERATURE AND AVIAN BLOOD ON LEISHMANIA DEVELOPMENT IN SAND FLIES TWO SAND FLY GENOMES: PHLEBOTOMUS PAPATASI AND LUTZOMYIA LONGIPALPIS Votypka J, Pruzinova K, Hlavacova J, Volf P Mary Ann McDowell and the Sand Fly Genome Consortium Department of Parasitology, Faculty of Science, Charles University, Prague, Czech Republic Eck Institute for Global Health, Department of Biological Sciences, University of Notre Dame, USA [email protected] [email protected] Keywords: temperature, blood, leishmania development Keywords: Lutzomyia longipalpis, Phlebotomus papatasi, genome Ambient temperature is one of external factors affecting pathogen transmission by haematophagous arthropods. Recent findings of new leishmaniasis foci in Latin America and Europe and the spread of the disease connected with the climate changes require more data on the effect of temperature on Leishmania development in sand flies. Therefore, the development of three important Leishmania species (L. infantum, L. braziliensis and L. peruviana) in two sand fly vectors (Phlebotomus perniciosus and Lutzomyia longipalpis) was compared at two different ambient temperatures (20°C and 26°C). At various intervals post-blood meal, infected females were examined under a light microscope and qPCR was used for parasite quantification. Leishmania peruviana, a mountain species living in lower temperatures, thrived well in Lu. longipalpis females kept at 20°C, while at 26°C almost all infections were lost due to fast digestion and early defecation of the blood remnants. On the other hand, L. braziliensis and L. infantum developed well in sand fly females kept at both temperatures tested. The ability to develop at low ambient temperatures could be an important factor enabling the spread of L. braziliensis and L. infantum into new areas and the risk of spread of both studied Leishmania species to higher latitudes and altitudes should not be over looked. Another factor affecting Leishmania development in vectors is the blood meal digestion by midgut proteases. Previously, some studies have demonstrated that avian blood inhibits Leishmania infection in the Old World vectors. However, this effect has never been observed in the New World sand flies. Therefore, our study was focused on the effect of chicken vs. rabbit blood on sand fly digestion and the development of Leishmania major in its natural vector P. duboscqi. In addition, we tested the effect of avian blood on midgut trypsin activity and the influence of repeated feedings on the susceptibility of sand flies to Leishmania infection. Sand fly females fed on chicken blood had significantly lower midgut trypsin activity and delayed egg development compared to those fed on rabbits. On the other hand, there was no effect of avian blood on parasite development within the sand fly gut. Similarly, blood feeding of sand flies on birds or mammals prior to the infectious meal did not cause any differences in Leishmania infections. Our study proved that sand fly feeding on avian hosts is not harmful to Leishmania parasites. The goal of the Sand Fly Genome Sequencing Project was to sequence the genomes of two different phlebotomine sand flies, Phlebotomus papatasi and Lutzomyia longipalpis, that exhibit distinct distributions, behavior, and pathogen specificity. Funded by United States National Human Genome Research Institute the Sand Fly Genome Project was initiated by a proposal submitted by Drs. Mary Ann McDowell, Frank Collins, Marcelo Ramalho-Ortigao, Jesus Valenzuela, Shaden Kamhawi, Rod Dillon, Paul Bates, and Michael Lehane. The Lu. longipalpis portion of the project is headed by Dr. Rod Dillon, Lancaster University, with sequencing at Baylor College of Medicine Sequencing Center, USA. The Ph. papatasi portion of the project is headed by Dr. Mary Ann McDowell, University of Notre Dame, USA with sequencing at Washington University Sequencing Center, USA. The genome of Lu. longipalpis (Jacobina strain) is approximately 150 Mb at 38.9x coverage. The N50 of the Lu. longipalpis contigs is 7.5 kb and the N50 of the scaffolds is 85.1 kb. The genome of Ph. papatasi (Israeli strain) was sequenced at 22.5x coverage and is approximately 350 Mb with a scaffold N50 of 23.6 kb. Genome assemblies of both species are available on VectorBase (https://www.vectorbase.org). Other genomic resources available from this project include a 40,000 ESTs from a normalized cDNA library of Ph. papatasi generated from the four larval stages, pupa, male and female at 1, 3, and 10 days post emergence and females post feeding on uninfected and Leishmania major infected mouse blood. RNAseq data is also available for these same conditions. In addition, the transcriptomes of Lu. longipalpis larval stages and females fed with sugar and uninfected or infected (Le. infantum) blood meals were sequenced. Genomic DNA from males and females from both species was used to generate individual BAC libraries. Further, SNP discovery is ongoing from individual flies collected from different geographical areas. Manual annotation is ongoing, but nearing completion. The following gene groups are being assessed: genes involved in digestion and immunity; circadian rhythm; microRNAs; G-protein coupled receptors, odorant and gustatory receptors; salivary protein genes, heat shock protein genes, cuticular protein genes, hormonal signaling, antioxidants, aquaporins, genes involved in symbiotic interactions, transposable elements, and potential viral sequences. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 62-O Genome & Genetics ID 63-O Genome & Genetics THE DATABASE VECTORBASE AND THE GENOMES OF LUTZOMYIA LONGIPALPIS AND PHLEBOTOMUS PAPATASI MANUAL ANNOTATION OF G PROTEIN–COUPLED RECEPTORS (GPCRs) IN THE GENOMES OF LUTZOMYIA LONGIPALPIS AND PHLEBOTOMUS PAPATASI Gloria I Giraldo-Calderón, Scott J. Emrich, Daniel Lawson, Frank H Collins and the members of the VectorBase consortium 53 Samuel Tadros, Mariha Wadsworth, Mallory Hawksworth, RJ Nowling, Mary Ann McDowell, Gloria I Giraldo-Calderón University of Notre Dame, European Molecular Biology Laboratory, EMBL - European Bioinformatics Institute, EBI, Institute of Molecular Biology and Biotechnology, IMBB, Harvard University, Imperial College London, The University of New Mexico Department of Biological Sciences, Eck Institute for Global Health, University of Notre Dame, Notre Dame, IN 46556, USA. [email protected] [email protected] Keywords: genomics, bioinformatics, database, genome sequencing Keywords: G protein–coupled receptors (GPCRs), gene manual annotation, new insecticide targets VectorBase (www.vectorbase.org) is a Bioinformatics Resource Center, which hosts the genomes of invertebrate vectors of human parasites and pathogens, including mosquitoes, tsetse flies, ticks, lice, kissing bugs and two sandflies, Lutzomyia longipalpis and Phlebotomus papatasi. Scientists working with sandflies in the field and/or the laboratory can become VectorBase users. The data deposited in VectorBase and in the public repositories, (such as GenBank/NCBI, EMBL/EBI or the DDBJ), are constantly updated and are a resource that has been subject to only very limited analysis to date. These data are freely available for new analyses, descriptions and hypothesis testing. Both public and private data can be queried or analyzed with a variety of available tools including VectorBase Blast, ClustalW, Hmmer, BioMart, Galaxy, Web Apollo and the Genome, Ontology, Expression and Population Biology browsers. The overarching goal of VectorBase is to support efforts to improve or develop new vector control or eradication strategies. Questions about how to use VectorBase, or suggestions about how to improve this resource, are very welcome, please send your comments to [email protected]. Sequencing the genomes of Phlebotomus papatasi and Lutzomyia longipalpis will help in the task to combat sand fly-borne diseases such as leishmanaisis. These sequences will provide valuable resources for improving control or eradication strategies to stop the transmission of these diseases. One such strategy is the use of G protein-coupled receptors (GPCRs) as new targets for insecticide chemistries. GPCRs play roles in many essential insect pathways and can be specifically targeted with small molecules, making GPCRs good insecticide targets. We used bioinformatic approaches to identify GPCRs in the genomes of these two sand flies. A new classifier pipeline called Ensemble*, was created to identify candidate GPCR peptides from the P. papatasi and L. longipalpis genome at Vector Base using the set of known GPCR peptides from Aedes aegypti, Anopheles gambiae, Apis mellifera, Drosophila melanogaster, Homo sapiens, and Pediculus humanus. The identified sand fly GPCR sequences were aligned using BLAST against the non-redundant protein sequences (nr) in NCBI. Putative functions were assigned based on the best and the most informative BLAST hits, in most of the cases the latter was from D. melanogaster. The sand fly sequences were aligned with homologous genes, mostly from insects used in the classifier pipeline, using MultAlin. The sand fly gene models were manually corrected using the annotation tool Artemis. Identified and manually annotated GPCRs were used to search (tBLASTn) the sand flies genome scaffolds for additional GPCR genes. When available, the gene models were confirmed using transcript evidence from Expressed Sequence Tags (ESTs) and RNA sequencing (RNAseq). The genes were named according to nomenclature guidelines developed for invertebrate vectors as detailed at VectorBase. If in doubt about the putative function of a gene, a literature search and a tree reconstruction was done using the software Clustal Omega. The same GPCR symbols used for other VectorBase genomes were also adopted, to facilitate the database query. The annotated gene models were submitted to VectorBase. Examples of conservation between homologous genes, lineagespecific gene duplications/deletions and the fragmented nature of the sand fly genome assemblies are also discussed. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 54 ID 64-O Genome & Genetics ID 65-O Genome & Genetics GENETIC VARIATION OF NYSSOMYIA TRAPIDOI (DIPTERA: PSYCHODIDAE) IN THREE ENDEMIC ZONES OF LEISHMANIASIS IN THE ECUADOR G E N E T I C A S S E S S M E N T O F S A N D F LY POPULATIONS IN PANAMA Gabriela Vela, Gabriel Trueba, Jérôme Depaquit, Sonia Zapata Anayansi Valderrama Cumbrera, Mara Garcia Tavares, Ingrid Murgas, José Andrade Dilermando Universidad San Francisco de Quito, Ecuador; Université de Reims Champagne Ardennes, France Instituto Conmemorativo Gorgas de Estudios de la Salud (ICGES), Universidade Federal de Viçosa, Centro de Pesquisas René Rachou [email protected] [email protected] Keywords: Ny. trapidoi, DNAmt, sympatry Keywords: Sandfly, cutaneous leishmaniasis, genetic population, genetic marker In the present study entomological collections were carried out between February and April 2013 in 3 provinces of Ecuador (Bolívar, Esmeraldas y Manabí). There were 14 species of sand flies identified, and Pressatia dysponeta, Psychodopygus panamensis Lutzomyia hartmanni and Nyssomyia trapidoi were the more abundant. Ny. trapidoi was found in 6 out of 7 points collected. The nucleotide analysis of the mitochondrial genes, cytochrome oxidase subunit I (COI) and cytochrome B (CytB) of 45 studied specimens allowed us to group this specimens in four clusters One of the clusters had specimens from all the studied provinces; meanwhile the other three had specimens captured only from Bolivar province. This results suggest that Ny. trapidoi is going through sympatric (inside Bolivar) and allopatric speciation (Between populations in Bolivar and the other provinces). Finally, it was found that 12,2% of the female samples analyzed were infected with Endotrypanum monterogeii. Many approaches have been implemented to understand the role of Phlebotominae sandfly in the dynamics transmission of leishmaniasis parasites. However, the introduction of genetic marker has allowed assessment of sandfly populations and the progress in the epidemiology understanding of leishmaniasis disease. In Panama, reports of human cutaneous leishmaniasis during the 60´s, 70´s and 80´s were attributed to sylvatic area and Lu. panamensis, Lu. trapidoi and Lu. gomezi as potential vectors. Since then, there have been modifications of the landscape, climate and an increase number of human cases is registered. In order to re-evaluate some criteria of the disease cycle aimed to study the genetic populations of Lu. panamensis, Lu. trapidoi and Lu. gomezi, therefore we estimate the genetic diversity, gene flow and demographic history of these species to infer the ability of survival and adaptability due change environmental. Individuals were collected with CDC light traps and identified to species level with taxonomic keys. DNA extraction of individuals were performed using phenol chloroform method and PCR amplification were implemented using the mitochondrial genes cytochrome oxidase I and the end region of the cytochrome b, intergenic region IGS -1, Ser - tRNA, IGS -2 and the initial region of the fragment NADH1 and intron elongation factor α-1. Preliminarily our results suggest that Lu. gomezi species showed a high genetic diversity and slight population structure may be due to the migration and local adaptation to environmental modifications by anthropogenic intervention. Knowledge of changes in the structure of Lutzomia genetics population may be helpful to understand the changes in the patterns of dynamics transmission of cutaneous leishmaniasis in Panamá. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 66-O Genome & Genetics ID 67-O Genome & Genetics EVIDENCE FOR GENETIC DIFFERENTIATION AT THE MICROGEOGRAPHIC SCALE IN PHLEBOTOMUS PAPATASI POPULATIONS FROM SUDAN GENE EXPRESSION PROFILING AND SILENCING OF KEY DIGESTIVE ENZYMES IN LUTZOMYIA LONGIPALPIS Noteila M Khalid, Marium A Aboud, Fathi M Alrabba, Dia-Eldin A Elnaiem and Frederic Tripet Department of Zoology, Khartoum College of Medical sciences, Khartoum, Sudan. 55 Hector M Diaz-Albiter, Caroline d Moraes, Maiara d Faria, Mauricio R Sant'Anna, Roderick J Dillon and Fernando A. Genta Fundação Instituto Oswaldo Cruz (FIOCRUZ), Brazil [email protected] [email protected] Keywords: digestion enzymes, RNAi, Lutzomyia Keywords: Phlebotomus papatasi, Sudan, Gene flow, Genetic differentiation. Background: Cutaneous Leishmaniasis (CL) is endemic in Sudan. It is caused by Leishmania major parasites and transmitted by Phlebotomus papatasi sandflies. Recently, uncommon clinical manifestations of CL have been reported. Moreover, L. donovani parasites that cause Visceral Leishmaniasis (VL) have been isolated from CL lesions of some patients who contracted the disease in Khartoum State, Central Sudan with no history of travelling to VL endemic sites on south-eastern Sudan. Because different clinical manifestations and the parasite behaviour could be related to genetic differentiation, or even sub-structuring within sandfly vector populations, a population genetic study was conducted on P. papatasi populations collected from different localities in Khartoum State known for their uncommon CL cases and characterized by contrasting environmental conditions. Methods: A set of seven microsatellite loci was used to investigate the population structure of P. papatasi samples collected from different localities in Khartoum State, Central Sudan. Populations from Kassala State, Eastern Sudan and Egypt were also included in the analyses as outgroups. The level of genetic diversity and genetic differentiation among natural populations of P. papatasi was determined using FST statistics and Bayesian assignments. Results: Genetic analyses revealed significant genetic differentiation (FST) between the Sudanese and the Egyptian populations. Within the Sudanese P. papatasi populations, one population from Gerif West, Khartoum State, exhibited significant genetic differentiation from all other populations including those collected as near as 22 km. Conclusion: The significant genetic differentiation of Gerif West P. papatasi population from other Sudanese populations may have important implication for the epidemiology of leishmaniasis in Khartoum State and needs to be further investigated. Primarily, it could be linked to the unique location of Gerif West which is confined by the River Nile and its tributaries that may act as a natural barrier for gene flow between this site and the other rural sites. The observed high migration rates and lack of genetic differentiation among the other P. papatasi populations could be attributed to the continuous human and cattle movement between these localities. Lutzomyia longipalpis is the most important vector of American Visceral Leishmaniasis. Adults of both sexes feed on sugars, while adult females are also haematophagous and larvae feed on solid detritus. Information regarding digestion in sand fly larvae is scarce. However, glycosidase activities involved in microorganism digestion have been described. In this work, β-1,3-glucanases, chitinases and digestive lysozymes were identified. Tissue-specific expression profiling was performed and physiological effects of RNAi-mediated silencing of these enzymes were explored. This is the first study to identify the molecular nature of sand fly larval digestive enzymes as well to assess their relevance by reverse genetics. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 68-O Virus Infections ID 69-O Virus Infections EVALUATION OF COLLECTION METHODS FOR PHLEBOTOMUS-BORNE VIRUSES DETECTION: I S O L AT I O N A N D V I R A L R N A I N T E G R I T Y PERFORMANCE CONDITIONED MEDIA AND EXOSOMES FROM LUTZOMYIA LONGIPALPIS LL5 CELLS DSRNATRANSFECTED ARE ABLE TO INDUCE NONSPECIFIC ANTIVIRAL RESPONSE IN LL5 NAIVE CELLS Remoli ME1, Bongiorno G2, Fortuna C1, Marchi A1, Bianchi R2, Khoury C2, Ciufolini MG1, Gramiccia M2 1 Unit of Viral diseases and attenuated vaccine and 2Unit of Vector-borne Diseases and International Health, MIPI Department, Istituto Superiore di Sanità, Rome, Italy [email protected] Keywords: Phlebotomus-borne viruses, Phlebotomus perniciosus, Toscana Phlebovirus, Experimental infection 56 Andrea Martins da Silva1, Antonio Jorge Tempone1, André N Pitaluga1, Fabricio K Marchini2, Barbara Burleigh3, Yara M Traub-Cseko1 1 Laboratório de Biologia Molecular de Parasitas e Vetores Instituto Oswaldo Cruz FIOCRUZ Pavilhão Leonidas Deane, Sl. 304. Av. Brasil, 4365. Manguinhos. Rio de Janeiro, RJ – Brasil, CEP 21040-360 Tel.: +55 21 3865-8206 / Fax: +55 21 25903495; 2Laboratório de Genômica Funcional Instituto Carlos Chagas / FIOCRUZ-PR R. Prof. Algacyr Munhoz Mader, 3775 CIC Curitiba – PR – Brasil, 81350-010 Phone +55 41 3316 3236 Fax +55 41 3316 3267; 3Department of Immunology and Infectious Diseases 665 Huntington Avenue Building 1, Room 817 Boston, Massachusetts 02115 Phone: 617.432.2495. [email protected] Phlebotomus-borne (PhB) viruses are RNA viruses involved in human disease outbreaks. Because several emerging viruses have been recently isolated from sand flies, an important task is to monitor the spreading of PhB-viruses. In order to investigate on collection and storage methods of phlebotomine vectors for reliable RNA detection and virus isolation, experimental viral infections of colonized sand flies were carried out and different laboratory conditions were tested. Laboratory-reared Phlebotomus perniciosus was experimentally infected with Toscana Phlebovirus (TOSV, ISS-Phl3 strain) using a membrane feeding technique. After the infected blood meal, engorged females were stored by two methods: 1) alive sand flies (AS) were immediately frozen at –80°C, mimicking the golden standard CDC light trap collection procedure; 2) insects were put onto sticky traps left at room temperature, collected daily up 144 h and stored dry at 80°C. Batches were separated into dead sand flies contaminated by alcohol necessary for removal from papers (DS-A) and those in which contamination was avoided (DS). The specimens, collected individually or in pools, were homogenized and used both for virus isolation in VERO cells and genome identification by Reverse Transcription-PCR on N gene region, and Real Time-PCR on TOSV L region. TOSV isolation percentage and RNA detection was 100% for the AS specimens, as expected. In DS specimens the TOSV isolation on VERO cells was elevated at 24 h but less satisfactory thereafter, suggesting that the virus could be fairly isolated from infected wild specimens only if they were frozen 24 h from death. No TOSV isolation was obtained from DS-A specimens at any collection time. As regards RNA detection, for all conditions examined Real TimePCR gave positive results until to 6 days after the infection. Recent investigations have indicated that virus diversity in the Mediterranean basin is higher than initially suspected, therefore a standardization of reliable sand fly collection and storage methods is important to harmonize entomological surveys. We confirmed that live specimens are the best target for both viral isolation and RNA identification. However sticky traps are suitable for partial genome viral identification but less so for viral isolation. This study was carried out in the frame of EDENext, a collaborative project of the 7th FP (20112014) funded by the European Commission under the DG Health; Contract Number: 261504. Keywords: antiviral response, conditioned media, exosomes Phlebotomine sand flies are haematophagous insect vectors of leishmaniasis in the New and Old World. In the Old World these insects are also related to virus and bacteria transmission. Although there is no proven scientific data on the New World sand flies capacity to transmit viruses, these insects may be considered as potential disseminators of these pathogens. The Lutzomyia longipalpis is the principal vector of visceral leishmaniasis in the Americas. Previous results of our group identified a nonspecific antiviral response in cultured L. longipalpis embryonic cells line LL5, in response to transfection with double stranded RNA (dsRNA). This was the first report of this kind of response in an insect cell line. We are presently interested in identify the mechanisms by which LL5 cells recognize dsRNA and trigger the antiviral response. Recently the role of secreted molecules in the host cells and pathogens has increased dramatically. In this work we incubate naive cells with conditioned media or isolated exosomes from dsRNA transfected and mocktransfected (control) LL5 cells, then infected these treated cells with West Nile virus-like particles expressing luciferase. Interestingly the cells incubated with conditioned media or exosomes from dsRNAtransfected cells, displayed a statistically significant antivirus response when compared with control groups. Because of this result, we decided study the protein composition of each one of these conditioned media (24 and 48 hours after transfection) and from exosomes isolated from these media. Samples from both sources were analyzed using mass spectrometry methodologies. Initial analysis identified 306 peptides in the conditioned media and 530 in exosomes. Some of these proteins were associated with antiviral response and/or structural vesicles. Among these proteins we can cite the Scramblase, FKBP, Adaptor protein and Nascent polypeptideassociated complex. These proteins were highly incident in exoproteoma from cells 24 hours after transfection, however, exhibited a reduction when analysed the exoproteome in 48 hours after transfection when we compared with control. The transcriptional pattern of some of these proteins was investigated in new LL5 cells cultures dsRNA-transfected. This is a first report on the exoproteome analysis related to antiviral response in phlebotomine embryonic cells. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 70-O Physiology & Colonization ID 71-O Physiology & Colonization CD-ROM SAND FLY FELLAS: THE FIRST COMPLETE VIDEO-BASED SAND FLY REARING GUIDE IN HIGH DEFINITION COMPARISON OF VOLATILE COMPOUNDS EMITTED BY ANIMALS OR PRESENT IN PLANTS ON ATTRACTIVENESS OF NYSSOMYIA NEIVAI (DIPTERA: PSYCHODIDAE) Meneses C, Morales E, Rhodes C, Oliveira F, Valenzuela JG and Kamhawi S. Vector Molecular Biology Section, Laboratory of Malaria and Vector Research, NIAID, NIH, Rockville, Maryland, USA. [email protected] 57 Vicente Estevam Machado1; Thais Marchi Goulart2; Arlene Gonçalves Corrêa3; Mara Cristina Pinto1 Faculdade de Ciências Farmacêuticas-UNESP (Brasil)1; Instituto de Biologia UNICAMP (Brasil)2; Departamento de Química -UFSCar (Brasil)3 [email protected] Keywords: Sand Fly, Video Based, Rearing guide, Leishmaniasis Keywords: primary alcohols, kairomone, wind tunnel, attractiveness It is well known that less than a hand full of sand fly species can be reared in the laboratory in sufficient numbers to provide enough material for large-scale experiments. Large-scale sand fly colonization is challenging and requires knowledge of sand fly biology, availability of the appropriate infrastructure and equipment, and having access to up to date protocols. With the resurgence of leishmaniasis, a neglected tropical disease, and the demonstration of the virulence of vector-transmission compared to needle-delivered parasites, significant investments have been made by institutions and companies to evaluate new leishmaniasis vaccines using infected sand flies. Development of experimental models of natural transmission requires a transition from small sand fly colonies to large-scale colonies. The objective of this presentation is to provide an account of the steps required to grow a large-scale sand fly colony and how to prevent or overcome problems that commonly arise during colonization. Our sand fly insectary unit, a branch of the Laboratory of Malaria and Vector Research (LMVR) at the National Institutes of Health, has acquired expertise in mass rearing of two major sand fly species: Lutzomyia longipalpis from Jacobina, Brazil and Phlebotomus duboscqi from Mali, Africa providing an annual productivity averaging 1.2 million reared sand flies and an experimental consumption of about 120,000 flies. With the goal of contributing to the sand fly community, we are sharing our expertise with other insectaries and interested laboratories through a 2.8 GB Windows© based CD-ROM with videos in high image definition embedded on a friendly user auto run menu designer™. The CD-ROM also contains sand fly rearing techniques according to our most up to date protocols; a complete list of materials needed to rear sand flies along with schematic drawings (AUTO CAD©) of custom-made items indispensable for handling a large number of flies on a daily basis with precision; a comprehensive list of references on sand fly rearing with links to PubMed; a link to Bei Resources where laboratories in need can request sand fly eggs or larvae of available vector species to initiate their own colonies; and a link to our lab where we can offer further advice on how to mass rear sand fly species of interest. Chemical ecology approach is a feasible tool for monitoring and maybe with intensive studies controlling species of haematophagous insects. Usually the studies are concentrated in the search for volatiles emitted from hosts which attract haematophagous insects (kairomones). Considering that males and females sand flies require sugar meal to survive, plants compounds can also be an interesting source for chemical ecology researches. In this study, we aimed to test the sand flies attractiveness in wind tunnel using two kairomones from hosts used in previous studies for haematophagous insects (lactic acid and 1-octen-3-ol) and primary alcohols that are usually found in many types of plants: 1- propanol, 1-butanol, 1-pentanol, 1hexanol, 1-heptanol, 1-octanol, 1-nonanol e 1-decanol.The tests in the wind tunnel were carried out with Nyssomyia neivai, an important vector of cutaneous leishmaniasis in South America. Groups of three female sand flies were placed inside releasing chambers in a total of 30 insects for each test and positioned 50 cm far from the stimulus. At the beginning we tested 200 µL in filter paper of each compound in three different concentration 10%, 50%, 100% (neat), if necessary the concentration of 5% was also included. Tests lasted two minutes and activation (number of insects that leave the release chamber) and attraction (number of insects that reach the stimulus) behaviors were evaluated. For the control group we used the diluents: hexane for alcohols and ethanol for lactic acid. Sand flies responded in a dose dependent way to 1-butanol, 1-pentanol, 1-hexanol, 1-octanol and 1octen-3-ol. The best responses for both behaviors evaluated were obtained with 1-hexanol, 1-octanol and 1-octen-3-ol. Lactic acid presented a weak response of activation and no attraction response. This result is in accordance with mosquitoes studies which have shown an activation role for lactic acid. The results showed that compounds from plants, like primary alcohols, may be an useful source for searching new compounds for trapping sand flies. ID 72-O Physiology and Colonization N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina RESPONSES OF THE SAND FLY PHLEBOTOMUS PAPATASI SCOPOLI (DIPTERA: PSYCHODIDAE) TO AN OVIPOSITION PHEROMONE ASSOCIATED WITH CONSPECIFIC EGGS STUDY ON THE BACTERIAL MIDGUT MICROBIOTA ASSOCIATED TO IMMATURE STAGE OF GENUS LUTZOMYIA (DIPTERA: PSYCHODIDAE) ISOLATED FROM NATURAL BREEDING SITES. Ifhem Chlebi1-2, Elyes Zhioua1, and Gordon Hamilton2 Vivero J Rafael1, 2; Cadavid Gloria3; Moreno H Claudia Ximena3; Posada-López Laura 1, 2; Sandra Uribe S. 1, 2 1 Institut Pasteur de Tunis, Laboratory of Vector Ecology, Tunis, Tunisia; 2Centre for Applied Entomology and Parasitology, Keele University, Stafordshire, UK [email protected] Keywords: Phlebotomus papatasi, oviposition pheromone, eggs. The chemical ecology of Lutzomyia longipalpis the South American vector of Leishmania chagasi infantum has been extensively studied and this species complex is known to produce both sex and oviposition pheromones. By contrast very few studies on the pheromones of Old World sandflies have been performed. Our preliminary studies have shown that female P. papatasi produce an oviposition pheromone. Female Phlebotomus papatasi laid a significantly (T-test; n=12 ; P <0.01) greater number of eggs on sites with 160 eggs already present than on control sites with no eggs. When smaller numbers of eggs (40 or 80) were used no significant differences between the mean numbers of eggs laid on test and control sites were detected. Furthermore when extract was prepared in hexane from batches of 160 eggs and placed on filter paper in test pots a significant increase in the number of eggs laid compared to the controls was found (Paired T-test, n=8, P < 0.05). Together these results indicate the presence of an oviposition pheromone on the eggs of P . papatasi. ID 73-O Physiology & Colonization 58 1 Laboratorio de Sistemática y Biología de Insectos (Insectario), Grupo de investigación en Sistemática Molecular, Facultad de Ciencias, Universidad Nacional de Colombia, sede Medellín; 2PECET (Programa de Estudio y Control de Enfermedades Tropicales Unidad de Entomología Médica y Molecular, Universidad de Antioquia; 3Grupo de Microbiodiversidad y Bioprospección. Universidad Nacional de Colombia-Sede Medellín [email protected] Keywords: immature, digestive tract, bacterial communities Background: Few researches provide information about the bacterial communities associated with the digestive tract of genus Lutzomyia and their possible role in the dynamics of transmission of the Leishmaniasis. The development of immature stages of genus Lutzomyia can include great variety of sites that contain decomposing organic material, where larvae may acquire different bacterial communities. The identification of such bacteria is of importance because their potential uses in designing improved vector control strategies. Bacterial communities could regulate the reproduction, abundance, distribution of Phlebotomine and can help identify the breeding sites of genus Lutzomyia. This premise has motivated new initiatives directed to study the bacterial communities associated with the digestive tract of immature individuals of genus Lutzomyia (found in natural breeding sites). Materials: Wild populations of immature stages of genus Lutzomyia, were collected in Ovejas (department of Sucre) in the Caribbean region of Colombia. The digestive tract of larvae and pupae were dissected. Digestive tracts were spread in a simple broth for enrichment. Genomic DNA was isolated from each of the pure cultures and amplified (PCR-Colony) the intergenic region of the 16S-23S rDNA and of the 16S gen which was sequenced to each isolate. The resulting sequences were compared with the GenBank database using the BLAST algorithm and software RDP. Phylogenetic affiliations of culturable bacteria and a NeighborJoining (NJ) dendrogram were constructed and the relevance of bacteria isolated from the gut was observed. Results: Our results indicated that there is a dominance of gram-positive bacteria in the digestive tract and the Malpighian tubules of immature individuals of genus Lutzomyia. Different ITS profiles were observed among immature stages (larvae and pupae). These results revealed a different flora in immatures of genus Lutzomyia (Streptomyces cinnabarinus, Ochrobactrum anthropi, Shinella zoogloeoides, Microbacterium pseudoresistens, Bacillus cereus, Paenibacillus illinoisensis, Lysobacter soli), when compared with other studies that only contemplate the adult stage of phlebotomine sand flies. Conclusion: This work provided basic information about bacterial diversity in midgut of immature stages of genus Lutzomyia. Some bacterial species are ideal targets that could be used for control. ABSTRACT BOOK POSTERS ISOPS VIII Puerto Iguazú Argentina N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 1-P Control ID 2-P Control SANDFLY FAUNA (DIPTERA: PSYCHODIDAE) OF URBAN AREA IN THE CITY OF SANDFLY FAUNA (DIPTERA: PSYCHODIDAE) FROM REGIONAL OF HEALTH IN SINOP, MATO GROSSO, BRAZIL SINOP, MATO GROSSO, BRASIL, YEAR 2013 Sirlei Franck Thies1,2,3, Ana Lucia Maria Ribeiro3,5, Jorge Senatore Rodrigues Vargas3,5, Edelberto Santos Dias4, Amilcar Sabino Damazo1 Sirlei Franck Thies1, 2, 3, Marinês Browers2 1 Laboratory of Histology, Department of Basic Sciences in Health, Medicine College, UFMT, Cuiabá/MT. 2Regional Office of Health of Sinop, State Department of Health of Mato Grosso, Sinop/MT. 3Laboratory of Medical Entomology, Medicine College, UFMT, Cuiabá/MT. 4Laboratory of Leishmaniasis, Center of Research René Rachou/Fiocruz-Minas Gerais. 5University Hospital Júlio Müller, UFMT, Cuiabá/MT. 60 1 Laboratory of Histology, Department of Basic Sciences in Health, Medicine College, UFMT, Cuiabá/MT. 2Regional Office of Health of Sinop, State Department of Health of Mato Grosso, Sinop/MT. 3Laboratory of Medical Entomology, Medicine College, UFMT, Cuiabá/MT. [email protected] [email protected] Keywords: Leishmaniasis, Sandfly, Lutzomyia Keywords: Leishmaniasis, Sandfly, Lutzomyia Mato Grosso State registers cases of autochthony of American Cutaneous Leishmaniasis (ACL) in 100% of its cities, and Sinop is one of the three cities in spotlight in number of cases. For Visceral Leishmaniasis (VL), a growing number of cities are registered with reports of autochthonal cases for this harm, a fact that worries health authorities, since they registered a human case in 2010 in Sinop. Sinop is located in the northern of Mato Grosso and has been a spot on the national stage for its socioeconomic growth, making it a commercial, educational and health polo. This study aimed to diagnose the sandfly fauna in the urban area in the city of Sinop, Mato Grosso. Entomological collections were made using CDC light traps, which were installed at 6 p.m. and collected at 7 a.m. the next day, during three nights in a row, in November 2013, around the homes located in 24 neighborhoods in the urban area of Sinop. The captured insects were prepared and identified in the Laboratory of Entomology of the State Department of Health of Mato Grosso. Thirty-three (33) specimen of sandflies, 17 male (51.55%) and 16 (48.5%) female, were captured. The captured sandflies belonged to 13 different species. The main species was Lutzomyia davisi (21.2%), followed by Lutzomyia antunesi (18.2%). Half of the neighborhoods were positive for sandflies. Other important species in the epidemiology of ACL were diagnosed, but at lower frequencies, such as: Lutzomyia whitmani (main vector for Mato Grosso) and Lutzomyia yuilli yuilli. The diagnosis of Lutzomyia whitmani, main vector of ACL in the state of Mato Grosso and Lutzomyia antunesi, suspected species for transmitting ACL and Visceral Leishmaniasis (VL), already found naturally infected by Leishmania chagasi in Mato Grosso, highlights the risk and the possibility of autochthonal transmission of ACL and VL in the city, even in urban areas. We emphasize the need for entomological and epidemiological monitoring, as well as the study of natural infection of these insects by Leishmania, studies which also involve the canine seroprevalence and the diagnosis of Leishmania circulation in humans, so that actions of prevention, promotion and/or control toward to the harm can be triggered, in order to minimize the contact of man/vector and the reduction in the number of cases in the city. The Leishmaniasis are zoonotic diseases caused by protozoa of the order Kinetoplastida, family Tripanostomatidae, genus Leishmania, transmitted by insects of the genus Lutzomyia, called sandflies, popularly known as white wing, stiff wing, birigui or straw mosquitoes. This study aimed to diagnose the current sandfly fauna in cities with reported cases of American Cutaneous Leishmaniasis (ACL) in Regional of Health in Sinop, Mato Grosso, during the year 2013. To this, entomological surveys were conducted from July to October 2013, with assistance from CDC light traps, installed at 6:00 p.m. and collected at 7:00 a.m. the following day, for three nights in a row. Insects captured were sent to the Laboratory of Entomology of the State Department of Health of Mato Grosso, where they were prepared and identified according to Young and Duncan, 1994. The Regional of Health in Sinop has 14 cities in its coverage area, making it possible to have the research done in only 5 cities in the period. A number of 3.029 sandflies were captured; the city with the largest number of insects captured was Nova Ubiratã 2.618 (86.43%). There was a predominance of male insects 1.985 (65.53 %). The city with the greatest diversity of caught species was Nova Ubiratã with 23 distinct species. The most prevalent species was Lutzomyia lenti 1.208 (40.77 %). In all cities species of importance in the epidemiology of ACL were diagnosed as: L. flaviscutellata and L. whitmani (main vector in Mato Grosso), as well as in Nova Ubiratã it was diagnosed L. longipalpis, vector of Visceral Leishmaniasis (VL). The diagnosis of the presence of vector species of ACL and VL lead us to recommend preventative and protective measures ranging from environmental management (sanitation) to activities of promotion and protection of health that should be developed along with the community, keeping a focus at minimizing human contact with the vector. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 3-P Control ID 4-P Control TESTING THE VERSATILITY OF A PSYCHODIELLA SP. DIAGNOSTIC ASSAY IN KINETICS OF ANTI-PHLEBOTOMUS PERNICIOSUS SALIVA ANTIBODIES IN MICE FIELD SURVEYS AND RABBIT SERA Lorena G. Caligiuri1,2, Enrique A. Sandoval3, María Soledad Santini4, Soraya Acardi5, Oscar Inés Martín-Martín, Ricardo Molina, Maribel Jiménez D. Salomón6, Lilian Tartaglino3 and Christina B. McCarthy1,2 61 Unidad de Entomología Médica, Servicio de Parasitología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Ctra. Majadahonda-Pozuelo s/n, 28220, Majadahonda, Madrid, Spain. 1 Centro Regional de Estudios Genómicos, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Argentina.2 Departamento de Informática y Tecnología, Universidad Nacional del Noroeste de la Provincia de Buenos Aires, Pergamino, Buenos Aires, Argentina.3 Laboratorio de Vectores, Secretaría de Calidad de Vida, Municipalidad de Posadas, Posadas, Misiones, Argentina.4 Laboratorio de Biología Molecular Aplicada, Facultad de Ciencias Exactas, Químicas y Naturales, Universidad Nacional de Misiones, Posadas, Misiones, Argentina.5 Centro Nacional de Diagnóstico e Investigación en Endemoepidemias, Administración Nacional de Laboratorios e Institutos de Salud, Ministerio de Salud, Buenos Aires, Argentina.6 Instituto Nacional de Medicina Tropical, Ministerio de Salud de la Nación, Puerto Iguazú, Misiones, Argentina. [email protected] Keywords: Psychodiella chagasi, diagnostic assay, PCR, field survey Psychodiella chagasi natural infections have previously been recorded in Lutzomyia longipalpis from Lapinha (Minas Gerais) (Adler and Mayrink, 1961), Jacobina (Bahia) in Brazil (Lantova et al., 2010) and, recently, Posadas (Argentina) (McCarthy et al. 2011). Given the fact that the Argentine Lu. longipalpis population is significantly differentiated from Brazilian populations (Salomon et al. 2010), we were interested in developing a straightforward and sensitive assay to analyse the incidence of natural gregarine infections in Lu. longipalpis from this location. In this context, we recently reported the first PCR-based assay for the detection and identification of natural gregarine infections in Lu. longipalpis (Caligiuri et al., 2014). Briefly, sand fly gregarine diagnostic primers PsyF and PsyR were designed on the basis of Ps. chagasi SSU rDNA sequences we previously identified in male Lu. longipalpis from Posadas (McCarthy et al. 2011). The specificity and sensitivity of the diagnostic primers were confirmed by in silico analysis and in vitro and field validations using total DNA extracted from naturally infected Lu. longipalpis (Caligiuri et al., 2014). Having validated our diagnostic assay for sand fly gregarine parasites in Lu. longipalpis, we were interested in testing its versatility for subsequent use in field surveys. Therefore, to evaluate the perfomance of the assay in a wider range of field captured Lu. longipalpis, we analysed 159 captured individuals from this species from Posadas (Argentina), using samples made up of one sand fly each and pools of 5 and 10 sand flies using different protocols and conditions. As expected, to obtain consistent results for the samples made up of one sand fly each, it was necessary to optimise the DNA extraction protocol. Our results indicated that the diagnostic assay is very versatile for field surveys and can be adapted to different sampling designs. Furthermore, in accordance with our previous results, the diagnostic assay detected gregarine infections that were not observed previously by light microscopy analysis. [email protected] Keywords: Phlebotomus perniciosus, anti-saliva antibodies, recombinant salivary proteins. Anti-saliva antibody response can be used as marker of exposure to sand flies. When hosts are bitten they develop humoral responses against sand fly saliva. Moreover, a positive correlation has been observed between number of bites and antibody levels. Little is known about kinetics of antibodies against salivary gland homogenate (SGH) or recombinant salivary proteins (rSP). This work focuses on the study of anti-Phlebotomus perniciosus saliva antibodies in sera of mice and rabbits that were experimentally exposed to the bites of uninfected P. perniciosus. In addition, IgG antibody levels against two P. perniciosus rSP, apyrase rSP01B (GenBank ID: KF178455) and D7 related protein rSP04 (GenBank ID: KF178456) were determined. Eight mice were immunized weekly through the bite of 150 sand flies for 12 weeks and their humoral response was followed by another 12 weeks. Six rabbits were weekly exposed to 500 sand fly bites for at least 4 weeks and their humoral responses were followed by 7 months. After nonexposure period both animals were exposed to test the presence of a B cell memory recall. All animals were bled according to the immunization schedule and anti-saliva antibodies were evaluated by ELISA. Anti-saliva response was also tested in rabbits by Western blot. In mice and rabbit sera, anti-saliva antibody levels increased along the immunizations and correlated with the number of sand fly bites. In both cases a memory response was highlighted. In mice, anti-SGH antibody levels were detected in sera 4 weeks after exposure, and persisted at least 3 months after the last exposure. Antiapyrase rSP01B antibodies followed similar kinetic responses that SGH, but with detectable antibody levels at fifth week. Anti-D7 protein rSP04 antibody response showed high variability among sera of immunized mice. In the case of rabbits, anti-saliva antibodies appeared after the 2nd week of exposure and antibodies were detected at 7 months post-exposure. In conclusion this study shows valuable data on sand fly saliva response in mice and rabbits. Results on kinetics of anti-saliva antibodies in rabbit sera provide useful information for an accurate application of anti-saliva response as marker of exposure in the field as wild rabbits have been proved to transmit Leishmania infantum to P. perniciosus in the focus of human leishmaniasis in south western Madrid region, Spain. Study supported by the Spanish Project AGL2008-01592 and by EU grant FP7-2011-261504 EDENext. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 5-P Control ID 6-P Control FIELD EVALUATION OF AN INSECTICIDE FORMULATED WITH ACTIVE INGREDIENTS ACTING ON LARVAL AND ADULT STAGES OF LUTZOMYIA LONGIPALPIS CHARACTERIZATION OF SUSCEPTIBILITY OF POPULATIONS OF LUTZOMYIA LONGIPALPIS ( D I P T E R A : P S Y C H O D I D A E ) TO ALPHACIPERMETRIN Laura W. Juan1, 2, Ignacio T. Gould3, Oscar D. Salomón4, Juan Molina4, Raul A. Alzogaray1, Eduardo N. Zerba1 Douglas de Almeida Rocha, Thaís Oliveira Coelho, Rodrigo Gurgel Gonçalves, Andrey José de Andrade & Marcos Takashi Obara 1 Núcleo de Medicina Tropical, Universidade de Brasília, Campus Universitário Darcy Ribeiro, Asa Norte, 70910-900, Distrito Federal, Brasil CIPEIN–Centro de Investigaciones de Plagas e Insecticidas (UNIDEF-CONICET), Villa Martelli, Buenos Aires. Argentina. 2Cátedra de Zoología. Facultad de Ciencias Agrarias. UNLZ, Buenos Aires. Argentina.3CeNDIE –Centro Nacional de Diagnóstico e Investigación en Endemoepidemias. Buenos Aires.REDILA 4 INMeT –Instituto Nacional de Medicina Tropical, Puerto Iguazú, Misiones. Argentina.REDILA [email protected] Keywords: Lutzomyia, chemical control, permethrin, pyriproxyfen In Argentina, the American tegumentary leishmaniasis (ATL) is endemic, with wild enzootic cycles, while visceral leishmaniasis (VL) is urban. VL is more severe because it can affect internal organs with high fatality. Since 2006, this disease is an emerging phenomenon of public health importance. The most practical way of prevention is to control the insect vector, phlebotomines sand flies from genus Lutzomyia, by application of residual insecticides, and dogs treatment with veterinary formulations. Recently, insecticide treated nets were introduced with controversial results. The application of insecticides in phlebotomines cities foci could be an important vector control tactic to reduce urban transmission. With the objective to develop a new control alternative, we evaluated a formulation containing the pyrethroid insecticide permethrin (10%) and the insect growth regulator pyriproxyfen (2%). According studies performed at our laboratory, permethrin is effective on adult L. longipalpis. Pyriproxyfen is an larvicide that disrupts metamorphosis of immature stages of sand fly, preventing it from reaching the adult stage. The trial was carried out in the city of Puerto Iguazú, Misiones, Argentina in March-April, 2014. Three dwellings with presence of L. longipalpis, all of them with poultry houses, were selected. Before applying the formulation, population level of sand fly were monitored for two days with CDC type traps. The formulation was emulsified in water and applied in two dwellings (TP and Chan). The third house (Ct) was untreated (control). The application was performed during sunset using a hand pulverizer. Application covered the interior of houses and surroundings to a distance of three meters. One week before the application the average of number of adults sand flies catched by the traps were 13 and 22 in the houses TP and Chan, respectively. Just before the treatment the flies catched during two days were 21 and 11, respectively. One month after the treatment, no flies were catched in TP and just one in Chan. In the control house, more than 50 sandflies were catched in all cases. This succesfull result could be attributed to the complementary effects of the insecticides applied: permethrin against adults insects and pyriproxyfen against larvae present in soil. 62 [email protected] Keywords: Sand flies, control, insecticides, visceral leishmaniasis Introduction: Visceral Leishmaniasis (VL) is an important public health problem in many countries of Latin America. Nowadays, none safe and effective vaccine exists and the transmission control of VL depends combating insect vectors using application of residual insecticides. Currently, there are records of outbreaks of sand fly populations resistant to DDT and BHC, in Sri Lanka, India and Venezuela. In Brazil, information about resistance of populations of Lu. longipalpis are limited and little is known about the genetic potential of this important resistance transmitter LV kind of man. Objective: Characterize the susceptibility profile of populations of Lu. longipalpis alfacipermetrina the pyrethroid used in Vector Control Program in Brazil. Methods: Dose-response bioassays with laboratory specimens (control) will be carried out by comparison them field populations from four endemic for VL in Brazilian states. Ten specimens of Lu. longipalpis will be exposed in bottles impregnated with insecticide in four alphacipermetrin concentrations to estimate the curve versus dose of insecticide mortality of insects. A total of 80 adult of sand flies (40 males and 40 females), F1 generation, between three to five days of life will be divided into four vials tests and one control. Reason of Resistance and Slope will be calculated for all populations. It will be considered from 98 to 100% mortality; as indication of susceptibility from 80 to 97%strength confirmation; and less than 80% mortality resistance will point established. All tests shall be repeated at least three times for each population on different days. Final considerations: The results will allow for the rapid and early detection of populations with probable evidence of insecticide resistance, contributing to the “Programa de Controle de Leishmaniose Visceral (PCLV)” of the Ministry of Health of Brazil in the deployment of alternative vector control strategies, as well as the periodic monitoring and systematic different populations of Lu. longipalpis, coming from different areas of the country. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 63 ID 7-P Control ID 8-P Control EFFECTS OF LEISHMANIA MAJOR INFECTION ON SAND FLY FEEDING RESPONSES TO DEET AND OR PERMETHRIN TREATED UNIFORM MATERIAL SOIL ANALYSIS AND VEGETATION ASSOCIATED WITH THE BREEDING SITES OF LUTZOMYIA EVANSI IN THE MUNICIPALITY OF OVEJAS-SUCRE A FOCUS OF LEISHMANIASIS IN COLOMBIA Tobin Rowland, Silas Davidson, Kevin Kobylinski, Edgar Rowton Walter Reed Army Institute of Research, Silver Spring, MD, USA [email protected] Horacio Cadena1, Edgar Ortega2, Luz A. Acosta1, Andrés Vélez-Mira1, Rafael J. Vivero1, Eduar E. Bejarano2, Sandra Uribe3, Hernando Gómez4, Luis G. Estrada2 & Iván D. Vélez1 1 Keywords: DEET, Permethrin, Leishmania major, Phlebotomus duboscqi Despite the use of Personal Protective Measures (PPMs) and other vector control applications, cases of tropical diseases like malaria, leishmaniasis, dengue, and other viral diseases still occur in deployed military service members. Some of these pathogens may affect the feeding behavior of the insects they infect. In sand flies, Leishmania parasites attach to the stomodeal valve of the anterior midgut. This attachment often interferes with the sand fly's ability to obtain a blood meal which can result in multiple probes. The objective of this study was to determine if Phlebotomus duboscqi infected with Leishmania major parasites responded differently than uninfected sand flies to commonly used PPMs; DEET applied to the skin and permethrin treated uniform material. A hairless guinea pig model was used to determine differences in the blood feeding rate and probing activity. Twenty sand flies were allowed to feed in a modified cage placed on anesthetized guinea pigs that had been treated with each PPM. DEET was applied to the skin at the rate recommended for people and permethrin treated uniform fabric was placed on the guinea pigs with a 8.75cm2 hole exposed. The Fisher's Exact test was used to compare the proportions of blood fed sand flies within and between treatment groups. Both the DEET and permethrin treated uniforms decreased the ability of sand flies to successfully take a blood meal, and there was no significant difference between infected and uninfected flies. When permethrin treated uniforms were used without DEET, there was a significance increase in the amount of probing activity by both infected and uninfected flies. This suggests that permethrin treated clothing may force sand flies to exposed skin and may actually increase the risk of disease transmission if not used with a topical skin repellent. PECET (Programa de Estudio y Control de Enfermedades Tropicales), Universidad 2 de Antioquia, Medellín, Colombia. Grupo de Investigaciones Biomédicas, Universidad de Sucre, Sincelejo, Colombia. 3Grupo de Investigación en Sistemática Molecular, Universidad Nacional, Medellín, Colombia. 4Atención Integral en Salud (ATIS)- Corposucre [email protected] Keywords: Leishmaniasis, Lutzomyia, breeding site, vegetation Several studies have shown that the physical and chemical characteristics of soil and vegetation are essential factors that determine the breeding sites of sand flies. This study was conducted in the peri-urban area of the municipality of Ovejas-Sucre, Colombia a endemic focus of cutaneous and visceral leishmaniasis. This region has two contrasting seasons: a rainy period from May to November and a dry period from December to April. The annual precipitation and mean temperature is 1380 mm and 28°C, respectively, and according to Holdridge the ecological life-zone is Tropical Dry Forest. The aim of this study was to describe the composition of soil and vegetation associated with the breeding sites of immature forms of Lutzomyia (Lu.) evansi the main vector of cutaneous and visceral leishmaniasis in the Colombian Caribbean region. Soil samples (n=43) were collected as follows: zone 1 (n=13), zone 2 (n=14) and zone 3 (n=16). In the three zones evaluated an average pH value of 7.43 (range 7.10-7.68) indicated a marginally basic soil. Exchangeable aluminum (Cmol. kg-1 soil) and salinity was zero. The texture of the samples was classified as sandy loam and silt loam soils. The organic content of the soils, which is important for larval development, was ̅x 3.35% (range 0.76 -5.83) being on average higher in the zone 1. In the three zones studied a total of 27 plots of 100 m2 each were sampled to characterize the vegetation associated with positive breeding sites. The vegetation in zone 1and 2 was characterized by herbaceous and shrub species with scattered trees. In contrast, in zone 3 a greater number of trees were recorded. Larvae (n=98 Lu. evansi) were predominantly recovered from the trees Cordia dentata, Guazuma ulmifolia and Manguifera indica. These results suggest that the knowledge of the peri and extradomiciliary breeding sites of sand flies could help orient environmental intervention measures that would affect the breeding sites of sandflies. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 9-P Control ID 10-P Control THE USE OF SYSTEMIC INSECTICIDES TO CONTROL PHLEBOTOMUS SPP. SAND FLIES IN SOUTHERN KAZAKHSTAN TRANSMISSION OF LEISHMANIA INFANTUM TO LUTZOMYIA LONGIPALPIS BY HUMANS WITH AND WITHOUT HIV INFECTION, SYMPTOMATIC AND ASYMPTOMATIC David Poché¹, Alex Gendernalik¹, Larisa Polyakova¹, Richard Poché¹, Madina Kozhahmetova², Batirbek Aimakhanov², Ziyat Abdeliyev², Bakhit Atshabar² ¹Genesis Laboratories, Inc.: Colorado, United States. ²Kazakh Science Center for Quarantine and Zoonotic Diseases by State Committee of National Sanitary-andEpidemiological Oversight of the Ministry of Health by Masgut Aikimbaev (KSCQZD): Almaty, Kazakhstan 64 Carlos C B Ribeiro, Antônio Menezes Filho, Daniela M Parente, Teresinha C Farias, Danielle A Zacarias, Jalthon C da Silva, Dorcas L. Costa, Ivete L Mendonça, Carlos H N Costa Universidade Federal do Piauí. [email protected] [email protected] Keywords: Leishmania infantum, Lutzomyia longipalpis, reservoirs, xenodiagnosis Keywords: Rhombomys opimus, Paraphlebotomus mongoliensis, fipronil, Bakanas Cutaneous leishmaniasis is a parasitic disease that causes ulcers on exposed parts of the body and leads to lifelong scars. The World Health Organization estimates that 0.7 to 1.3 million new cases of CL occur annually, making it the most common form of leishmaniasis. In the Old World, the vector for CL is the Phlebotomus spp. sand fly. The primary reservoir host in Central Asia is the Great Gerbil (Rhombomys opimus). Phlebotomus spp. sand flies were discovered in Bakanas, Kazakhstan for the first time in June 2012. In light of this discovery, it was determined that a field trial should be conducted to test the efficacy of the insecticide, fipronil against Phlebotomus spp. sand flies by feeding it to great gerbils in bait form. A field trial was conducted in Bakanas, Kazakhstan in May and June 2013 to determine the abundance of Phlebotomus spp. sand flies in the area. In addition, the efficacy of a fipronil-based bait in controlling female sand fly numbers was to be conducted. The study design involved monitoring sand flies using CDC light traps for two weeks prior to treatment and for four weeks post treatment on two treatment plots and two control plots. Both treatment plots were treated with a one-time application of fipronil based gerbil bait. Sand flies that were collected from CDC light traps were sexed and identified down to the genus level for Sergentomyia spp. and to the species level for Phlebotomus spp. Data for total numbers of sand flies per trap night were recorded in an Excel Spreadsheet. Two weeks of additional CDC light trapping were performed in September 2013. The research resulted in some interesting ecological findings. Nearly 100% of the sand flies collected were Paraphlebotomus mongoliensis. Of particular interest was the density of blood fed female sand flies. Of the 3,487 female P. mongoliensis that were collected, 1,248 had taken blood meals (36%). This number is abnormally high and the data could be very beneficial to developing a sand fly control program in Kazakhstan. The efficacy of the fipronil bait is yet to be determined. Differently from visceral leishmaniasis (VL) caused by Leishmania donovani, which is a disease transmitted only among people, VL caused by L. infantum is a zoonosis transmitted from domestic and wild canines. However, patients with VL coinfected with HIV or immunocompetent are also known to infect the vectors Phlebotomus perniciosus and Lutzomyia longipalpis. Besides, Lu. longipalpis is an eclectic vector, including to humans, who are important blood sources for this species in urban settings in Brazil. Therefore, nevertheless neglected by control programs, the role of humans as reservoirs of L. infantum seems plausible. This study aimed to compare the competence of symptomatic and asymptomatically infected humans, with and without HIV, as reservoirs of L. infantum. The study groups were composed by (a) 21 patients with LV and HIV (VLHIVs); (b) two asymptomatic patients with LV and HIV (VLHIVa); (c) 21 patients with VL but without HIV (VLs); and (d) 19 asymptomatic individuals infected with L. infantum but without HIV (VLa). All patients underwent xenodiagnosis with Lu. longipalpis. Insects were dissected and examined by optical microscopy and by conventional PCR for Leishmania spp. The infections ratios were 13/21 in VLHIVs; 1/2 in VLHIVa; 8/21 VLs: and 3/19 in VLa. The results show that being symptomatic for VL increases infectivity of humans, but also that asymptomatic individuals with or without HIV infection are also able to transmit L. infantum to the vector Lu. longipalpis. These results demonstrates that humans should be considered as source of L. infantum when designing control programs for urban VL in the New World. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 11-P Control ID 12-P Ecology and epidemiology ADDRESSING THE EMERGENCE AND SPREAD OF LEISHMANIASIS IN THE BORDERS OF ARGENTINA, BRAZIL AND PARAGUAY MOLECULAR DETECTION OF LEISHMANIA IN PHLEBOTOMINE SAND FLIES IN A CUTANEOUS AND VISCERAL LEISHMANIASIS ENDEMIC AREA IN NORTHEASTERN BRAZIL Quintana María Gabriela1 ,2,.3 ,4, Santini María Soledad1 ,2 ,5, GonzálezBritez Nilsa6, Rojas de Arias Antonieta7, Celeste Vega8, Thomaz-Soccol Vanete9, Bisetto Júnior Alceu10, Yadon E Zaida11, Salomón Oscar Daniel1,2,4 and the IDRC Project #107577 team. Instituto Nacional de Medicina Tropical-MSN, Argentina; 2 REDILA,Argentina; 3 Instituto Superior de Entomología-UNT, Argentina; 4 CONICET, Argentina; 5Centro Nacional de Diagnóstico e Investigaciones en Endemoepidemias-ANLIS-MSN, Argentina; 6 Facultad de Ciencias de la Salud-Universidad Nacional del Este, Paraguay; 7 OPS/OMS Paraguay, 8 Centro para el Desarrollo de la Investigación Científica (CEDIC), Asunción, Paraguay. 9 Departamento de Engenharia de Bioprocessos e Biotecnologia, Univerasidade Federal do Parana, Brazil; 10 Secretaria de Saúde do Estado do Paraná, Brazil, 12 Communicable Diseases Department, Pan American Health Organization, Washington, D.C., United States of America 65 Vanessa Cristina Fitipaldi Veloso Guimarães, Pietra Lemos Costa, Fernando José da Silva, Fábio Lopes de Melo, Filipe Dantas-Torres, Eduardo Henrique Gomes Rodrigues and Sinval Pinto Brandão-Filho 1 Keywords: Eco-Health, borders, multidisciplinary In the border region of Argentina, Brazil and Paraguay, cutaneous leishmaniasis generates rural and peri-urban outbreaks strongly associated with environmental and anthropogenic change Facultad de Ciencias de la Salud-Universidad Nacional del Este, Paraguay.s, while visceral leishmaniasis is emergent and spreading in urban and periurban areas. Therefore, different forms of transmission of leishmaniases occur in a diversity of settings, ranging from forest to urban, and from work to peri-domestic associated risks. The region also has a wide diversity of human populations, including different cultural backgrounds, as native peoples, migrants (transient and permanent settlements), and tourists. In order to address the emergence and spread of leishmaniasis a project was designed to identify the main biological, environmental and social drivers associated with parasite transmission risks in different landscapes, within the theoretical frame of Eco-Health. This project engages researchers and decision makers from the three countries to respond to the explicit demand of public health authorities to improve the effectiveness of leishmaniasis control programs. The design involves “mirror” activities in Argentina, Brazil and Paraguay, and iterative/successive feedbacks instead of simultaneous analysis between the results of the dialectic sciences (social) and cumulative ones (bio-ecological). Up to now the project team consolidated the historical information about the leishmaniases, and the sociohistorical context that allowed the transmission in the region, the social scientists performed a base-line perception study, and the three countries multidisciplinary specialists gathered in a workshop to establish a common epistemological background and to refine the field designs. The antecedents included the previous studies on distribution in time and space of Phlebotomine, and its relation with outbreaks and/or humans activities. In each country a specific set of interventions will be developed. The participation of decision makers from the local, subnational and national levels, the technical collaboration of the Pan-American Health Organization and the partnership of the International Development Research CentreCanada, will promote the uptake of results and implementation of the proposed multi-sectorial prevention and control strategies, as well as their eventual replication in other areas. Financial Support IDRCCanada Project #107577, Ministry of Health of Argentina, Brazil Paraguay; PAHO. Centro e Pesquisas Aggeu Magalhães- Fiocruz [email protected] Keywords: Phlebotomine sand flies, Leishmania; transmission Several phlebotomine sand fly species have been regarded as putative or proven vectors of parasites of the genus Leishmania in Brazil, but data for the northeastern region remains incipient. In this study, a total of 600 phlebotomine sand flies were grouped in pools of 10 specimens each and tested by a Leishmania genus-specific PCR and by a PCR targeting Leishmania (Leishmania) infantum. Fourteen out of 60 pools were positive by the genus-specific PCR, being five pools of Lutzomyia migonei, seven of Lutzomyia complexa, one of Lutzomyia sordellii and one of Lutzomyia naftalekatzi, which corresponds to a minimal infection rate of 2.3% (14/600). Our results, associated with their known anthropophily and their abundance, suggest the participation of L. migonei and L. complexa as vectors of Leishmania in northeastern Brazil. Remarkably, the detection of Leishmania DNA in L. sordellii and L. naftalekatzi is reported for the first time in this country, but future studies are necessary to better understand the significance of these findings. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 13-P Ecology and epidemiology ID14-P Ecology and Epidemiology STUDY OF SAND FLY FAUNA IN RECENT AREA TRANSMISSION OF LEISHMANIASIS IN THE STATE OF MINAS GERAIS, BRAZIL N E W R E C O R D A N D U P D AT E GEOGRAPHICAL DISTRIBUTION OF MONTICOLA (COSTA LIMA, 1932) PSYCHODIDAE: PHLEBOTOMINAE) AMERICA Menezes, Júlia Alves1; Sousa, Alessandra Mara2; Morais, Mayron Henrique Gomes2; Andrade-Filho, José Dilermando1; Lima, Fernanda Pinheiro3; Fux, Blima4; Margonari, Carina1 1 2 66 ON THE PINTOMYIA (DIPTERA: IN SOUTH Andrey José de Andrade & Rodrigo Gurgel-Gonçalves Centro de Pesquisas René Rachou, Brazil; Fundação Educacional de Divinópolis, Brazil; 3Centro Universitário de Formiga, Brazil; 4Universidade Federal do Espirito Santo, Brazil Laboratório de Parasitologia Médica e Biologia de Vetores, Área de Patologia, Faculdade de Medicina, Universidade de Brasília, Campus Universitário Darcy Ribeiro, Asa Norte, 70910-900, Distrito Federal, Brazil. [email protected] [email protected] Keywords: Lutzomyia longipalpis, Lutzomyia cortelezzii, Lutzomyia whitmani, Visceral Leishmaniasis Keywords: Pintomyia monticola, potential distribution, Federal District, ecological niche modeling Introduction: Study the sand fly fauna in recent foci of leishmaniasis is crucial to know the risk of disease transmission, since it allows to clarify the distribution and vectors ecology. In the city of Formiga, Minas Gerais, Brazil, the first cases of American visceral leishmaniasis (AVL) occurred in 2011 and 2012, although still have not been recorded american cutaneous leishmaniasis (ACL) cases. Thus, the present study aimed to know the local sand fly fauna and identify potential areas of transmission risk. Methodology: Monthly systematic collections in the 24 peridomicile houses, using HP light traps, ocurred for 4 consecutive days (5pm - 7am) between May 2012 to April 2013. The domiciles were selected in accordance with the location of human and canine cases of leishmaniasis and the peridomicile conditions. The insects captured were identified by Young and Duncan key (1994). The Pearson correlation and chisquare tests were used to test the association of phlebotomines with bioclimatic factors (humidity, temperature and rainfall). Results: Out of 224 specimens were captured from 10 species, the most prevalent were Lutzomyia longipalpis (n=79, 35.3%), Lutzomyia cortelezzii (n = 75, 33.5%) and Lutzomyia whitmani (n=41, 18.3%). The first two presented widely distributed by the municipality (22 and 18 collection points, respectively), while L. whitmani concentrated in just 4 points, especially near the green areas. A significant correlation between environmental conditions and abundance of sandflies were observed. It was found a high correlation between sandflies and rainfall (r2=0.52, p=0.008) and a moderate correlation between phlebotominaes and temperature (r2= 0.36, p=0.039). Conclusions: The occurrence of AVL cases and the finding of L. longipalpis indicate that probably this species is associated with the transmission of the leishmaniases in the city. This hypothesis agree with the fact that L. longipalpis was found in almost all collection points, but more studies are necessary. As for L. whitmani, even without records of ACL cases, their finding indicates the necessity of local entomological surveillance, due to the involvement of this species in the disease cycle . Finally, the near quarters with preserved forest areas, where the presence of L. whitmani were abundant, may exhibit a higher vulnerability to the appearance of the first cases of ATL. Pintomyia monticola was described based on five females captured in the state of São Paulo and males were described using specimens from a laboratory colony. The species have been recorded in Peru, Argentina, Paraguay and Brazil. It is a suspected vector of Leishmania enriettii, the etiological agent of cutaneous leishmaniasis in guinea pigs, in southern Brazil. The present study updates the distribution of P. monticola and presents a potential geographical distribution model for this species. Distributional data were obtained from various published studies and speciesLink data (http://splink.cria.org.br). When only textual georeferences were provided they were georeferenced based on internet resources (http://www.fallingrain.com/world/). Records were georeferenced with an uncertainty of ≤5 km to the nearest 0.01° (records presenting greater uncertainty were removed). Data were separated into two sets: one for model calibration (75% of points) and one for model evaluation (25% of points). Eight climatic variables from the WorldClim project were obtained to characterize climate variation across the distribution range of P. monticola. All environmental databases used in this study covered areas of South America, resample to a spatial resolution of 2.5' (~5 km). Ecological niche models were produced using Maxent version 3.2.1. We used median output grids as a hypothesis of suitability, and imported the results into ArcView 3.3 for analysis. We assessed model accuracy by examining omission rate associated with test points. We also used Maxent's jackknife test to identify variables that most influenced model predictions. P. monticola was registered in 143 localities and its updated geographical distribution map showed the bona fide records in South America and the new record (seven females) in Brasília, Federal District, Brazil. When niche models were displayed in geographical space P. monticola showed a wide potential distribution in Brazil passing through Bolivia and reaching Peru. All test points were included in the predicted suitable area (0% omission). Highest climate suitability for P. monticola was observed along the Brazilian Atlantic coast. When contribution of climatic variables was explored, annual mean temperature, temperature seasonality, and minimum temperature in the coldest month were the variables that most influenced models of P. monticola. The results presented here extended the geographical distribution of P. monticola in South America. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 67 ID 15-P Ecology and Epidemiology ID 16-P Ecology and Epidemiology E N TO M O L O G I C A L M O N I TO R I N G O F T H E PROGRAMME OF VISCERAL LEISHMANIASIS IN MACEIÓ, ALAGOAS STATE, BRAZIL: INDICATORS AND ENVIRONMENTAL RISK FACTORS ASPECTS OF THE ECOLOGY OF LUTZOMYIA LONGIPALPIS AND LUTZOMYIA MIGONEI (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) IN AREA OF INTENSE TRANSMISSION OF VISCERAL LEISHMANIASIS IN NORTHEAST OF BRAZIL: FORTALEZA, STATE OF CEARÁ, BRAZIL Carlos Fernando Rocha Dos Santos; Magliones Carneiro Lima; Grimoaldo Braga Da Rocha Neto; Ana Lúcia Moreira Macena Secretaria de Saúde de Maceió. Alagoas. Brazil [email protected] Rafaella A. Silva, Claudia M. L. Bevilaqua, Elizabeth F. Rangel, Fabricio K. M. Santos, Lindemberg C. Sousa. Lucas Edel Donato, Grasielle C. D. Pessoa Ministério da Saúde do Brasil Keywords: Maceió, Phlebotominae, Lutzomyia The city of Maceió is ranked, according to the methodology of the Ministry of Health as sporadic transmission to American Visceral Leishmaniasis (AVL). The occurrence of cases follows the process of human occupation, located in discontinuous areas and sparse, with half of the territory is comprised of rural areas, integrating even this scenario, a park and an area of environmental preservation. Entomological surveillance actions were triggered initially aimed to determine the distribution of the vector, Lutzomyia (Lutzomyia) longipalpis through entomological survey and monitoring, in order to know the seasonality. Monthly collections were made with the help of CDC light traps in the period from July 2004 to December 2012, and selected areas for implementation of entomological monitoring from identifying the points of greatest abundance of sand flies in the entomological survey, correlated to the presence of human and canine cases of the disease. The traps were installed from 18h and 07h drawn around the next day for three consecutive nights, totaling 20 traps indoors and peridodomestic sites. We collected 16,765 sand flies and recorded nine species, especially L. (Nyssomyia) whitmani (58%), L. (Lutzomyia) longipalpis (30%) and L. migonei (10%), all other species amounted to 2%. The locations of the Povoado Canto do Farias and Sitio São Gonçalo and Riacho Doce were those which had the highest abundance vector. Entomological indicators showed variation according to the presence of domestic animal shelters around the houses, and also regarding the conditions of residence and can be seen a decrease in vector abundance inside the house after improvements made. The results showed that there was a relationship between sand fly density and housing conditions and environmental factors, especially the presence of domestic animals (chickens and pigs). Environmental characteristics and lack of adequate infrastructure in communities, and how precarious proximity with the forest, presence of pets around the houses, garbage accumulation, favored the maintenance of sand fly vector breeding sites, increasing the risk of disease transmission. The entomological data found allies to environmental risk factors identified in the area suggest the need to consider in an integrated manner, entomological indicators and environmental management as a condition for effective control actions. [email protected] Keywords: Lutzomyia longipalpis, Lutzomyia migonei, Visceral leishmaniasis, Fortaleza The main vector for visceral leishmaniasis (VL) in Brazil is Lutzomyia longipalpis. However, the absence of L. longipalpis in a region of autochthonous VL demonstrates the participation of other species in the transmission of this disease. Studies conducted in La Banda, Argentina, and São Vicente Férrer, Brazil, have correlated the absence of L. longipalpis and the presence of L. migonei with autochthonous cases of VL. In São Vicente Férrer, there was evidence for natural infection of L. migonei with Leishmania infantum chagasi, which strongly supports the participation of this vector in disease transmission in the area. Thus, the objective of this work was to assess the ecology of the sand flies L. longipalpis and L. migonei in Fortaleza, an endemic area for VL. Insect capture was conducted at 22 sampling points that were distributed across four regions of Fortaleza, and two CDC light traps were used at each collection point. In total, 32,403 sand flies were captured; of these, 18,166 (56%) were identified as L. longipalpis and 14,237 (44%) as L. migonei. There were significant differences found between the vectors in each sampling site (indoors and outdoors) (p <0.0001). These findings confirm that L. migonei and L. longipalpis are distributed throughout Fortaleza, where they have adapted to an indoor environment. Because these species have been shown to be competent vectors for VL, our results suggest that L. migonei shares the role of vector with L. longipalpis for the transmission of VL in Fortaleza. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 17-P Ecology & Epidemiology ID 18-P Ecology & Epidemiology PHLEBOTOMINAE CAPTURED IN AREA OF TRANSMISSION OF AMERICAN CUTANEOUS LEISHMANIASIS (ACL) IN XAPURI MUNICIPALITY, ACRE STATE, WESTERN BRAZILIAN AMAZONIAN. PRELIMINARY RESULTS THE GEOGRAPHIC EXPANSION OF AMERICAN VISCERAL LEISHMANIASIS ASSOCIATED WITH DEFORESTATION IN TOCANTINS STATE, BRAZIL Andreia Fernandes Brilhante, Eunice Aparecida Bianchi Galati, Cristiane de Oliveira Cardoso, Nathanna Progênio dos Santos, Jailson Ferreira de Souza and Joaquim Vidal Faculdade de Saúde Pública da Universidade de São Paulo / Faculdade Meta Grupo Athenas Educacional [email protected] Keywords: Sand flies, vectors, american cutaneous leishmaniasis, Amazonia The study aimed identify the phlebotomine fauna in the Xapury municipality. The sand flies were captured monthly from August 2013 to February 2014 with automatic light traps (CDC type) from 6:00pm to 06:00am in rural and urban areas. Twelve traps were installed in forests, peridomiciles (in domestic animal shelters) and intradomiciles. The insects were macerated and identified in accordance with Galati (2003). A total of 3,064 specimens (1,065 males and 1,999 females) have been captured. They belong to 47 species: Bichromomiya flaviscutellata (0.07%), Brumptomyia sp (0.10%), Br. avellari (0.03%), Br. penthacanta (0.19%), Evandromyia bacula (0.03%), Ev. begonae (0.13%), Ev. infraspinosa (0.10%), Ev. saulensis (1.86%), Ev. termitophila (0.10%), Ev. walkeri (0.03%), Lutzomyia sherlocki (0.22%), Migonemyia migonei (0.03%), Nyssomyia antunesi (0.65%), Ny. richardward (0.07%), Ny. shawi (23.3%), Ny. whitmani (0.62%), Ny. yuillii yuillii (0.59%), Psathyromyia aragaoi (0.30%), Pa. campbelli (0.03%), Pa. cuzquena (0.03%), Pa. dendrophila (0.20%), Pa. punctigeniculata (0.03%), Pa. (Forattiniela) sp (0.10%), Pa. serie Shannoni (0.03%), Pintomyia nevesi (0.65%), Pi. serrana (0.03%), Pressatia sp (0.40%), Pr. choti (0.16%), Pr. tricantha (0.03%), Psychodopygus serie Guyanensis (0.62%), Ps. amazonensis (0.13%), Ps. carrerai carrerai (9.6%), Ps. claustrei (0.13%), Ps. davisi (9.14%), Ps. llanosmartinsi (8.25%), Ps. hirsutus hirsutus (2.61%), Ps. lainsoni (1.80%), Ps. paraensis (0.13%), Ps. yucumensis (0.35%), Sciopemyia sordellii (0.10%), Thrichophoromyia auraensis (10.20%), Th. octavioi (19.50%), Th. sp. (5.6%), Th. sp. n (0.13%), Th. ubiquitalis (1.40%), Trichopygomyia sp. (0.07%) and Viannamyia furcata (0.13%). About 95% of the all specimens collected belong to the three genera: Nyssomyia (25.2%), Psychodopygus (32.8%) and Trichophoromyia (36.9%). Four specimens of Trichophoromyia of a species not yet described were found. Of the two predominant genera Nyssomyia and Psychododpygus there are several anthropophilic species and some of them suspect of transmit ACL agents and Th. auraensis is the proved vector of Leishmania lainsoni. The occurrence almost exclusively of the sand flies in rural area with vector species suggest that the transmission of ACL agents has occurred in this environment. The residents and visitors should be reminded, especially because one of the investigated points has attracted tourists from the region and other areas including from abroad. 68 Margarete Martins dos Santos Afonso1, Sérgio Augusto de Miranda Chaves 2, Mônica de Avelar Figueiredo Mafra Magalhães 3, Renata Garrido 3, Carina Graser 41, Bruno M. de Carvalho1, Simone M. Costa 1, Elizabeth Ferreira Rangel 1 Laboratório de Transmissores de Leishmanioses, Laboratório de Referência em Vigilância Entomológica: Taxonomia e Ecologia de Vetores das Leishmanioses, Instituto Oswaldo Cruz Fundação Oswaldo Cruz; 2Laboratório de Ecologia, Departamento de Endemias, Escola Nacional de Saúde Pública Sérgio Arouca Fundação Oswaldo Cruz; 3Laboratório de Geoprocessamento, Instituto de Comunicação e Informação Científica e Tecnológica em Saúde; 4Secretaria de Saúde do Estado do Tocantins. [email protected] Keywords: Lutzomyia (L.) longipalpis, Epidemiology, Visceral Leishmaniasis, Spreading and Urbanization American Visceral Leishmaniasis (AVL) is an important Public Health issue in Brazil, with evident geographic expansion to urban areas of medium and large cities. Over the last decade, increasing levels of environmental changes have been recorded in Tocantins State, associated with agriculture and hydroelectric plant constructions. This scenario is observed in some regions of the state where AVL is present. Currently the disease is recorded in 120 of 139 municipalities. This study aimed to evaluate the AVL expansion in Tocantins State, by associating the deforestation increase with AVL stratification, as well as the presence of AVL with deforestation from 2004 to 2011. Annual deforestation rates in the Amazon region were obtained from the PRODES project (National Institute for Space Research, INPE). Disease stratification data was provided by the Health Department of Tocantins State. The software ArcGIS 10 was used for spatial analysis and map production. Chi square tests (confidence interval 95-99%) were applied with software SPSS 19 to evaluate the association between human cases of AVL and presence of deforestation. From 2004 to 2011, the state's northern region showed a constant increasing in deforestation and high numbers of AVL human cases, especially in the municipalities of Araguaína, Araguatins, Bandeirantes do Tocantins, Arapoema, Santa Fé do Araguaia, Ananás, Pau d'Arco, Itaporã do Tocantins, Nova Olinda, Colméia, Colinas do Tocantins and Riachinho. The results showed an association between deforestation and AVL transmission, with significant correlation values in 2005 (p=0.119), 2007 (p=0.114), 2008 (p=0.218), 2009 (p=0.039), 2010 (p=0.013), and 2011 (p=0.004). This confirms the hypothesis that the deforestation favors disease transmission. Most municipalities with AVL human cases had also deforestation records. The increase of the statistical significance over the last three years (2009-2011) is noteworthy. In Tocantins State, the analysis showed an association between deforestation and AVL transmission, corroborating studies which associate deforestation to epidemic outbreaks. In Brazil, AVL human cases are increasingly being recorded in environmentally changed areas. The results suggests the need for integrated planning and development of public policies to minimize the impacts of human-made environmental changes in AVL transmission. Financial support: IOC/FIOCRUZ, CAPES, CNPq (PAPES VI, 407707/2012-3), FAPERJ (E-26/110.525/2012). Edición especial (marzo 2015) Boletín de la Sociedad Entomológica Argentina ID 19-P Ecology & Epidemiology Financial support: IOC/FIOCRUZ, CAPES, CNPQ (PAPES VI, 407707/2012-3), FAPERJ (E-26/110.525/2012). LUTZOMYIA (LUTZOMYIA) LONGIPALPIS FEEDING HABITS AND NATURAL INFECTION BY LEISHMANIA (LEISHMANIA) INFANTUM CHAGASI IN SOME BRAZILIAN ENDEMIC AREAS OF AMERICAN VISCERAL LEISHMANIASIS ID 20-P Ecology & Epidemiology Margarete Martins dos Santos Afonso1, Sérgio Augusto de Miranda Chaves2, Daniela de Pita Pereira3, Taiana Amâncio da Costa Rego3, Antônio Luis Ferreira de Santana1, Júlia dos Santos Silva1, Bruno Moreira de Carvalho1, Simone Miranda da Costa1, Cristina Giordano4, Elizabeth Ferreira Rangel1 1 Laboratório de Transmissores de Leishmanioses, Laboratório de Referência em Vigilância Entomológica: Taxonomia e Ecologia de Vetores das Leishmanioses, Instituto Oswaldo Cruz - Fundação Oswaldo Cruz; 2Laboratório de Ecologia, Departamento de Endemias, Escola Nacional de Saúde Pública Sérgio Arouca Fundação Oswaldo Cruz; 3Laboratório de Biologia Molecular e Doenças Endêmicas Fundação Oswaldo Cruz, 4 Secretaria de Saúde do Estado do Rio de Janeiro. [email protected] 69 SAND FLIES (DIPTERA, PSYCHODIDAE, PHLEBOTOMINAE) COLLECTED IN THE WAJÃPI INDIGENOUS RESERVE, EASTERN AMAZON, BRAZIL Eduardo Stramandinoli Moreno1; Maíra Posteraro Freire2,4, Luis Otávio Cordeiro Nascimento3, Volmir Zanini3, Veracilda Ribeiro Alves4, Paloma Helena Fernandes Shimabukuro4 1 DSEI Amapá e Norte do Pará – Secretaria Especial de Saúde Indígena /Ministério da Saúde. 2ILMD–PPG Saúde, Sociedade e Endemias da Amazônia (UFAM/FIOCRUZAmazônia/UFPA) 3Laboratório Central de Saúde Pública do Amapá–LACEN/AP. 4 Centro de Referência Nacional e Internacional para Flebotomíneos-Centro de Pesquisas René Rachou/FIOCRUZ. [email protected] Keywords: Lutzomyia (L.) longipalpis, Leishmania (L.) infantum chagasi, Visceral Leishmaniasis, Ecology Brazil faces the geographical expansion and urbanization of American Visceral Leishmaniasis (AVL). The presence of the vector, Lutzomyia (Lutzomyia) longipalpis, in urban areas is one of the greatest challenges for Brazilian Visceral Leishmaniasis Control Program. The knowledge on the biology of L. (L.) longipalpis can help to develop more appropriate and effective control strategies as well as preventive actions. The study aimed to combine diagnostic molecular tools to detect, simultaneously, natural infection by Leishmania (Leishmania) infantum chagasi and the feeding source of L. (L.) longipalpis. Sand fly specimens were captured in the following Brazilian municipalities: Araguaína (Tocantins State), Fortaleza (Ceará State), Sobral (Ceará State) (the three with high incidences of AVL) and Rio de Janeiro (Rio de Janeiro State) (first urban focus of the city). Molecular diagnostics were performed by Polymerase Chain Reaction and Dot-Blot Hybridization with primers for Homo sapiens, Canis familiaris, Didelphis albiventris, Gallus gallus and L. (L.) infantum chagasi. The evaluation of the feeding source had a general positivity index of 29.3%. Lutzomyia (L.) longipalpis showed high positivity for humans (61.5%), followed by dogs (28.2%) and birds (10.3%). In Araguaína municipality, most of the samples were positive for humans (61.5%) followed by dogs (38.5%). In Fortaleza and Sobral, specimens had higher positivity for humans (54.5% and 66.7%), followed by dogs (27.3% and 20.0%) and birds (18.2% and 13.3%). The detection of feeding source from specimens collected in Rio de Janeiro was not possible, maybe because the specific primer was not used. The evaluation of natural infection had a general positivity index of 4.61%, with 4.9% from Araguaína and 9.75% from Sobral specimens. The simultaneous detection of feeding source and natural infection had the following results: in Araguaína, 2 specimens were naturally infected, one with dog blood and one with human blood; in Sobral, 3 specimens were naturally infected and had ingested human blood. Results corroborate previous demonstrations of the feeding plasticity and opportunistic behavior of L. (L.) longipalpis: it feeds on dogs, domestic reservoirs of L. (L.) infantum chagasi; on birds, animals whose attract sand flies and facilitate their domiciliation; and on humans, which reinforces its anthropophilic habit. Keywords: Amazon, Cutaneous leishmanises, Phlebotominae, Vectors To contribute to the understanding of the epidemiology of American cutaneous leishmaniasis (ACL) among indigenous people in Eastern Brazilian Amazon entomological collections were carried out at the Wajãpi Indigenous People Reserve in the state of Amapá, Brazil. The Wajãpi people consist of a group of approximately 1,000 indigenous people living in a remote area of the Brazilian Amazon forest, close to the border with French Guyana. The Wajãpi are subsistence agriculturalists who also hunt and collect forest products such as fruits and seeds. Life in the forest puts them in contact with the transmission cycles of ACL. Sand flies were collected during the dry season (September 2013) and rainy season (January 2014) with a total of 46 light traps distributed in 16 communities where cases of ACL have been reported. The traps were set from 16:00-07:00hs, in intra and peridomicile areas, and one area used frequently by the Wajãpi, such as rivers, gardens or hunting trails. A total of 6,150 sand flies have been collected. A total of 14 species have been identified so far, the most abundant species are Trichopygomyia trichopyga (56.3%), Psathyromyia dreisbachi (26.4%), Trichophoromyia ubiquitalis (3.8%), Bichromomyia flaviscutellata (3.8%), Evandromyia infraspinosa (3.4%) and Th. brachipyga (1.9%). The remaining species represented 4.4% of the total amount collected and they were: Nyssomyia umbratilis, Pa. aragaoi, Th. ininii, Sciopemyia sordellii, Sc. fluviatilis, Pa. inflata, Psychodopygus squamiventris maripaensis and Ty. depaquiti, the latter is recorded for the first time in Brazil. Three medically important sand fly species were collected Ny. umbratilis, Ps. squamiventris maripaensis and Bi. flaviscutellata. Sand flies were found in all areas sampled in both seasons suggesting ACL can be transmitted in the intra and peridomicile areas. In addition to sand fly collections, we have assessed social and environmental variables such as presence of wild and domestic animals, waste management, distance to rivers, gardens and forest areas, etc. which will help in the identification of ecological patterns for sand fly occurrence as well as the in the evaluation of hot spots for ACL transmission among the Wajãpi people. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 70 ID 21-P Ecology & Epidemiology ID 22-P Ecology & Epidemiology LUTZOMYIA LONGIPALPIS (LEISHMANIA INFANTUM VECTOR) AND NYSSOMYIA WHITMANI (LEISHMANIA BRAZILIENSIS VECTOR) IN THE CITY OF PUERTO IGUAZÚ, NORTHEASTERN ARGENTINA: SEASONAL ABUNDANCE AND RELATIONSHIP WITH DOMESTIC ANIMALS GEOGRAPHICAL DISTRIBUTION OF NYSSOMYIA BARRETTO, 1962 IN BRAZIL Caroline Amaral Machado, Veracilda Ribeiro Alves, Paloma Helena Fernandes Shimabukuro Centro De Pesquisas Rene Rachou/Fiocruz Minas Fernández María Soledad ; Santini María Soledad ; Cavia Regino ; and Salomón Oscar Daniel1,2 [email protected] REDILA 1Instituto Nacional de Medicina Tropical, 2Consejo Nacional de Investigaciones Científicas y Técnicas, 3Centro Nacional de Diagnóstico e Investigaciones en Endemoepidemias 4Dpto. de Ecología, Genética y Evolución, FCEN, UBA and Instituto de Ecología, Genética y Evolución de Buenos Aires UBACONICET. [email protected] Keywords: Cutaneous leishmaniasis, vectors, Nyssomyia umbratilis, Nyssomyia whitmani 1,2 3 4 Keywords: Sandflies, urban, peridomiciles, ecology In this study the city of Puerto Iguazú was sampled from spring 2011 to winter 2012 to evaluate seasonal abundance changes of sandflies. The city was divided in 51 areas of 400x400m where minilight traps were located at the beginning of every season during two consecutive nights in peridomiciles of households. Lutzomyia longipalpis and Nyssomyia whitmani represented the 97.5% of the captures (N=5409 sandflies). Two abundance indexes were analyzed for each species: the occurrence and the number of individuals captured by night in each household. The seasonality of occurrence and abundance by species was evaluated using Generalized Linear Mixed Models. We also evaluate the effect of the number of dogs and chickens on the occurrence of sandflies at each household. For both analyses, household were considered as a random effect. Both species showed differences by season in the number of households-occurrence and number of individuals captured, but with different patterns. The percentage of households with Lu. longipalpis was higher in fall (66.7 %) compared to spring (31.4%) and summer (27.5%), and lower in the winter (10.2%). The percentage of households with Ny. whitmani was minimum in fall (9.8%) and was maximum in summer (39.2%) and spring (31.4%), while winter has an intermediate value (22.5%). The occurrence of both species was positively associated with the number of dogs and chickens in the households. For Lu. longipalpis also a synergic effect was detected in the number of dogs and chickens, while for Ny. whitmani the effect of the number of chickens varied among seasons. The percentage of households infested with both species was maximum at the spring (21.5%) and summer (23.5%). The number of individuals captured of both species changed according to the seasons similarly to the occurrence. Lu. longipalpis abundance was higher in fall (27.8 Phlebotomine per trap-night [PTN]) compared to spring and summer (5.6 and 5.3 PTN) and the winter was the season with the lower value (0.6 PTN). Ny. whitmani numbers were minimum in fall and in winter (1.0 and 0.4 PTN) and were higher in summer and spring (12.0 and 6.8 PTN). These results shows that in the study area these sandflies have different patterns of abundance according to the seasonality and also the numbers of domestic animals in the households as chickens or dogs, and so the seasonal and spaceenvironmental risk is different for cutaneous leishmaniasis and visceral leishmaniasis in the same city. The genus Nyssomyia Barretto, 1962 comprises 20 species of which 14 are recorded in Brazil, including vectors of Leishmania in American cutaneous leishmaniasis (ACL) transmission areas of Central and South America. The aim of this study is to map the geographical distribution of the species from the genus Nyssomyia in Brazil and discuss the distribution of medically important species. A dataset is being built based on literature search on PubMed, and records obtained from examination of specimens from two entomological collections: Coleção de Flebotomíneos of Centro de Pesquisas René Rachou and Museu de Zoologia da Universidade de São Paulo. So far, our results show the distribution of Nyssomyia anduzei in the north region of Brazil with most records from Amazonas state (AM), this species has been frequently associated with wild and semi-domestic areas in the north region; N. umbratilis has been mostly recorded from Pará state (PA), and more recently records for this species are from AM, mainly from Manaus city and its surroundings, this species has been found associated with humans and domestic animals accidentally when in contact with transmission cycles in the forest; N. delsionatali, N. richardwardi, N. shawi, N. sylvicola, N. urbinatti, N. yuilli pajoti and N. yuilli yuilli present more restricted geographical distribution in Brazil, mainly for the north and central regions of Brazil; N. singularis has been only recorded in São Paulo state (SP) from the type-locality suggesting it is not a valid species; N. neivai is recorded for the southeast region. Most records are for N. whitmani, in all regions of Brazil, especially in the southeast, including Minas Gerais state, where it has been found in as diverse environments as urban, rural and forest areas, In SP it has been recorded throughout the state. Nyssomyia antunesi has been found in both wild and semi-domestic areas in the north and northeast regions, mainly in AM and PA states. Nyssomyia intermedia is the most adapted species to anthropic environments, presents more records for Rio de Janeiro and Espírito Santo states, but is found in all regions of Brazil with the exception of the north region. Our compiled data contributes to the knowledge of the geographical range of Nyssomyia and shows where most work on ACL has been carried out by researchers and public health workers. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 23-P Ecology & Epidemiology ID 24-P Ecology & Epidemiology VERTICAL AND HORIZONTAL DISTRIBUTION OF SAND FLIES IN AN AREA OF TEGUMENTARY LEISHMANIASIS IN CHACO, ARGENTINA NATURAL BREEDING SITES OF PHLEBOTOMINAE (DIPTERA: PSYCHODIDAE) IN A FLOOD ZONE OF CHACO, ARGENTINA. Rosa JR1, Parras MA1,2, Szelag EA1,2 Salomón OD2 71 Parras Matías Ariel1, 2, 3, Rosa Juan Ramón2, 3, Szelag Enrique Alejandro1, 2, , Salomón Oscar Daniel1, 3 3 1 Laboratorio Entomología. Instituto de Medicina Regional. Universidad Nacional del Nordeste. Chaco. Nodo REDILA (Red de Investigación de la Leishmaniasis en Argentina). 2Instituto Nacional de Medicina Tropical (INMeT). Iguazú, Misiones. Argentina. 1 Instituto Nacional de Medicina Tropical (INMeT), 2Instituto de Medicina Regional (IMR - UNNE), 3Red de Investigación de la Leishmaniasis en Argentina (REDILA) [email protected] [email protected] Keywords: Stratification, Migonemyia migonei, Evandromyia complex , Bionomy During the active search for human cases of cutaneous leishmaniasis in a rural area (26º20'22"S, 60º15'44") of Tres Isletas a house was selected as worst epidemiological scenario. Five CDC-type light traps/month/night/12hs(19:00 to 7:00 pm) were used, in domicile, peridomicile (goats pens and piggery) and forest 110m from domicile where deforestation and anthropic activity were evident. Also in the forest two traps were installed in a tree, near the ground on 1.5 m and canopy level 10m. From December to December 2010-2012 a total 1179 Phlebotominae were captured: Migonemyia migonei (82%), Evandromyia cortelezzii (2.4%) only males, Ev. sallesi (5%) only males, Cortelezzii complex females (9%): (including Ev. cortelezzii and Ev. sallesi of similar morphology), Nyssomyia neivai (0.3%), Psathyromyia shannoni (0.1%) and Brumptomyia brumpti (1.2%). The most abundant was Mg. migonei on December 2011 (n=309) with average temperature and relative humidity of 31ºC and 32%, respectively. Only in summer 2010 (November/December) all species were recorded in peridomicile and domicile with the greatest abundance in the first (n=467) and the lowest in the second (n=81) predominating Mg. migonei in both ecotopes (n=405; n=71, respectively). In vertical strata, all species were recorded in ground level. In 2010, the highest abundance was canopy (n=289) in autumn (March/April), and the lowest in ground level (n=245) in summer (November/December). Migonemyia migonei was the predominant specie followed by Ev. cortelezzii, Ev. sallesi and Cortelezzii complex females. The least abundant were Pa. shannoni only in ground level and Br. brumpti in ground and canopy levels. During all seasons 2010 there was Phlebotominae in both strata, the highest abundance was in summer (n=422) with averages of temperature and relative humidity of 28°C and 45% and lower in winter (n=20), 12°C and 55%. In 2011, the highest was in spring (n=166) 24°C and 66% and the lowest in summer (n=74) 27°C and 58%. Species suspected as vectors of leishmaniasis in this region were recorded in this study in both strata in all seasons. This behavior was probably due to the meteorological and microclimate conditions, soil type and vegetation, limited by physiological factors such as changes in the food habits favored by anthropic activities. This highlights the need for further large-scale studies to contribute to the knowledge of bionomics and environmental change in an epidemiological context. Keywords: Nissomyia neivai, Lutzomyia longipalpis, soil samples, emergence traps Phlebotomine preimaginal sampling was performed from August 2011 to April 2014 in Puerto Antequeras (27° 26'34"S, 58° 50' 57" W) where adults of Lutzomyia longipalpis and Nissomyia neivai where already reported as the main vectors of visceral and cutaneous leishmaniasis, respectively. Two homes were selected, with the criteria of 'worst scenario'. From August 2011 to March 2013 substrate samples 10cm diameter by 3cm deep were taken monthly; and from April 2013 until April 2014 emergence traps modified from Casanova (2001) were installed. In each home three ecotopes were selected: domestic (external walls of the houses), peri-domestic (hen houses and kennels) and extra-domestic (base of trees and fallen logs and exposed roots). In the latter ecotope three points (trees) were selected at 100m distant from each other, along transect from the domicile. The collected soil samples were kept at 24±5ºC and 80±10% relative humidity and observed daily for 60 days in search of the emergence of imagos for identification of species level. After this period the remaining of the samples were processed by flotation with supersaturated solution of NaCl and observed under stereomicroscope looking for larvae, pupae, dead adults and/or pupal exuviuae. The emergence traps were controlled every 15 days to renew the sticky paper. Each trap remained in the soil for 60 days and then was moved to another point within the same ecotope and mictrohabitat considered as potential breeding site of sand-fly. A total of 950 substrate samples were taken. A male of Ny. neivai was obtained during flotation of a sample taken from the forest 100 meters from the house, at the base of a fallen tree in January (summer). As a result of the emergence traps 5 specimens of Ny. neivai were obtained (2 males and 3 females) distributed in four out of the forty traps. One was located in peridomestic ecotope (hen house) and three in the forest (base of a tree). Of the specimens obtained, 4 were in months before an extraordinary flood occurred between June and July 2013 and 1 was recovered nine months after that flood (April 2014). A production of 3.60 phlebotominae/100m2/day was estimated. These preliminary results showed the fitness of the floodplain area of the Parana River in the province of Chaco for natural breeding sites of Ny neivai. These studies are continued broadening the environmental range to support the hypothesis and looking for the breeding sites of Lu. longipalpis. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 72 ID 25-P Ecology & Epidemiology ID 26-P Ecology & Epidemiology ANALYSIS OF SAND FLY (DIPTERA: PSYCHODIDAE) BLOODMEALS USING SEQUENCING TECHNIQUE: TEST OF SAND FLY EXPERIMENTALLY FED SANDFLY SURVEY IN ECOTOURISM REGION IN THE DISTRICT OF BOM JARDIM FROM THE CITY OF NOBRES, MATO GROSSO - BRAZIL Carvalho GML; Quaresma PF; Gontijo CMF; Rego FD; Rugani JMN; Dias TA; Andrade Filho JD Anagela Maria Acel, Ana Lucia Maria Ribeiro, Rosina Djunko Miyazaki, Jorge Senatore Vargas Rodrigues Centro de Pesquisas Rene Rachou - Fiocruz Minas. [email protected] Universidade Federal de Mato Grosso [email protected] Keywords: Phlebotominae, Bloodmeal Molecular Biology The eco-epidemiological factors involved in the genesis of different forms of leishmaniasis in the New World are very complex. Several species of Leishmania are capable of causing disease in humans which in turn, interact with various sand fly species and mammalian hosts. The result of these interactions is a profuse epidemiological variety that difficult the understanding and control of the disease. Studies considering the occurrence of different species of Leishmania in sand flies have been essential for a better understanding of the ecoepidemiology of leishmaniasis. However, there are few studies investigating the correlation between infected sand fly species and their possible infection sources. Thus, the aim of this work is to standardize the bloodmeal identification in experimentally engorged females by sequencing technique, for future application in naturally engorged females. For this study, we selected as target a fragment of the Cytochrome B gene, present in birds and mammals and recently used in other studies to identify the meal in hematophagous insects. The tests were carried out using females of the first generation of a sand fly colony and were exposed to the first meal in different vertebrates. After the experimental fed, the females were sacrificed and subjected to DNA extraction and polymerase chain reaction technique (PCR) of the CytB target. Our results demonstrated a good amplification of the fragment of 359bp in experimentally fed specimens. The amplified products were purified and subjected to sequencing and it was possible to identify each particular level vertebrate host by aligning and comparing the sequences deposited in GenBank. These first results were crucial for the next steps that are being performed in attempt to optimize the technique and reproduce the methodology in naturally engorged females. The identification of the blood meal in naturally engorged females provides information about food sources and may improve the knowledge about the behavior aspects of sand flies like feeding preference or feeding patterns in natural conditions. These informations could also contribute to indicate the potential reservoir hosts, for Leishmania species in different ecotopes. Financial Support: FAPEMIG/CAPES, CPqRR/FIOCRUZ, CNPq. Keywords: Phlebotomine, Leishmaniasis, Nobres-Bom Jardim/MT, Ecotourism The Leishmaniasis are infectious diseases, non-contagious, chronic evolution caused by the protozoa of the genus Leishmania and transmitted by the bite of insects called sandflies which belong to the family Psychodidae, Genus Lutzomyia. The American Cutaneous Leishmaniasis (ACL) and Visceral Leishmaniasis (VL) primarily cause zoonotic infections, affecting humans and their pets. In Mato Grosso there is a great diversity of sandfly species with reporting cases in 100 % of the municipalities of ACL. The City of Nobres in the State of Mato Grosso / Brazil, situated 142 km from the capital of Mato Grosso, has natural wonders that attract more and more the interest of tourists, highlighting the district of Bom Jardim by numberless places for ecotourism. This study aimed to survey the insect fauna of sandflies in the district of Bom Jardim. CDC light traps were used, 8 (eight) indoor, 18 (eighteen) in peridomestic and 4 (four) outdoor, all installed in homes and places with tourist attractions in the months of December 2012 (starting rainy season); June/2013 (drought) and March/2014 (rainy season). A total of 465 sandflies were captured, divided into two genera and 14 species as it follows: Brumptomyia = 42 (9,0%); Lutzomyia antunesi = 171 (36,8%); L. aragaoi = 3 (0,6%); L. carmelinoi = 42 (9,0%); L. davisi = 30 (6,5%); L. evansi= 1 (0,2%); L. flaviscutellata = 5 (1,1%); L. hermanlenti = 22 (4,7%); L. lenti = 15 (3,2%); L. longipalpis = 3 (0,6%); L. walkeri = 116 (24,9%) e L. whitmani = 9 (1,9%). The proportion between male and female was 52.7 % and 47.3 % respectively. Regarding the collection environment, the percentage was: indoor=7.7%; peridomestic=87.5% and 4.7% outdoor. Among the three collection periods, the period that obtained a greater diversity of species and greater quantity with a total of 92.47% of occurrence, was December. Species that have implications in the transmission of leishmaniasis are: L. flaviscultellata, L. whitmani and L. antunesi to ACL and L. longipalpis to VL. Because this place is a great potential for ecotourism, it should be monitored and a program of preventive measures for leishmaniasis should be developed for both local people and tourists. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 27-P Ecology & Epidemiology ID 28-P Ecology & Epidemiology LEISHMANIASIS SANDFLY DIVERSITY IN A RURAL AREA OF THE CUIABÁ, MATO GROSSO, BRAZIL INCIDENCE OF SANDFLIES AT THE ZOOLOGICAL PARK OF FEDERAL UNIVERSITY OF MATO GROSSO, CUIABÁ/MT-BRAZIL Ana Lucia Maria Ribeiro1,2, Jorge Senatore Vargas Rodrigues1,2, Tiago Parreiras de Almeida2, Emerson Marques de Souza3, Flávia Karolina Pereira Barreto Bettiol3, Anagela Maria Acel1,2, Fabio Alexandre Leal dos Santos1,2, Sirlei Franck Thies1,2, Rosina Dunjko Miyazaki3 1 - Julio Muller University Hospital, Faculty of Medicine, UFMT, Cuiabá, Mato Grosso, Brazil 2 -Faculty of Medicine, UFMT, Cuiabá, Mato Grosso, Brazil 3 - Institute of Biosciences, UFMT, Cuiabá, Mato Grosso Brazil 73 Rosina Dunjko Miyazaki, Ana Lucia Maria Ribeiro, Jorge Senatore Vargas Rodrigues, Tiago Parreiras de Almeida, Emerson Marques de Souza, Flávia Karolina Pereira Barreto Bettiol, Anagela Maria Acel Universidade Federal de Mato Grosso [email protected] [email protected] Keywords: Vector, Leishmaniasis, Zoological Park, Cuiabá/MT Keywords: Phlebotomine, Leishmaniasis, Cuiabá/MT Leishmaniasis is an infectious disease caused by protozoa of the genus Leishmania, transmitted to mammalian hosts by insect vectors belonging to Lutzomyia genus. Most of the infections are observed among wild and domestic animals. Accidental zoonotic infections in man occurs mainly in tropical areas, including in Brazil. According to World Health Organization reports, endemic transmission of leishmaniasis occurs in 98 countries, however there is no official data for several others. The aim of this study was to investigate the variety of sandfly species in a rural area of Cuiabá, state of Mato Grosso (MT), Brazil.The study was conducted along the Fish river, located on the road linking Cuiabáto Chapada dos Guimarães,MT. CDC (Center for Disease Control) light traps where installed close to four residences at 18 PM and collected at 6 AM of the next day between June and November, 2013.Captured insects where stored in ethanol 70% and identified using Young and Duncan classification at the Entomology Laboratory. In this study, 29 Lutzomyia sandflies belonging to 10 species were captured. The most abundant was Lutzomyia cruzi (11/29; 37.93%), followed by Lutzomyia whitmani and Lutzomyia acanthopharyns (4/29;13.79% of each), Lutzomyia evandroi (3/29;10.34%), Lutzomyia longipalpis (2/29; 6.90%) and Lutzomyia punctigeniculata, Lutzomyia corumbaensis, Lutzomyia lenti, Lutzomyia lutiziana and Lutzomyia sp. (1/29; 3.45% of each).August and September were the months with the highest occurrence of sandflies. Lutzomyia whitmani (main vector of American Cutaneous Leishmaniasis in MT), Lutzomyia cruzi and Lutzomyia longipalpis (vector species of visceral leishmaniasis) were the most frequent sandflies found. This area will remain under entomological and epidemiological surveillance, considering the risk of cutaneous and visceral leishmaniasis transmission. Leishmaniasis is a group of diseases caused by protozoan of the genus Leishmania, which can be visceral or cutaneous manifestations that is caused by a protozoan and transmitted by an insect of the genus Lutzomyia, called sandflies. This study aims to monitor the presence of sandflies in the UFMT Zoological Park, for the control of vectors and adopt preventive measures in the transmission of leishmaniasis in animals and humans. Catches were made with CDC light trap in four points as farmed birds, small mammals, large mammals and reptiles. Samples were collected monthly for three consecutive nights from January 2012 to December 2013. 121 individuals of the genera Brumptomyia = 7 (5.8%) and Lutzomyia carmelinoi = 3 (2.5%) were captured; L. cruzi = 76 (62.8%); L. hermanalenti = 5 (4.1%); L. lenti = 7 (5.8%) L. whitmani = 20 (16.5%) and Lutzomyia sp = 3 (2.5%).The sex ratio was 67.8% males and 32.2% females. The months of highest incidence were June 2012 and 2013. Among the collected species L. cruzi is considered vector of visceral leishmaniasis and L. whitmani of American cutaneous leishmaniasis. The results indicate that the strategy of control and monitoring of the vectors must be continuous throughout the year. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 29-P Ecology & Epidemiology ID 30-P Ecology & Epidemiology P O P U L AT I O N D Y N A M I C S A N D H A B I TAT PREFERENCES OF PHLEBOTOMUS ORIENTALIS IN EXTRA-DOMESTIC HABITATS OF METEMA HUMERA LOWLANDS - KALA-AZAR ENDEMIC AREAS IN NORTHWEST ETHIOPIA ECOLOGY OF LUTZOMYIA LONGIPALPIS IN AN AREA OF VISCERAL LEISHMANIASIS TRANSMISSION IN NORTH-EASTERN BRAZIL Wossenseged Lemma1*, Habte Tekie2 Meshesha Balkew3, GebreMichael3, Alon Warburg4, Asrat Hailu5 74 Pietra Lemos Costa, Filipe Dantas-Torres, Fernando José da Silva, Vanessa Cristina Fitipaldi Veloso Guimarães, Kamila Gaudêncio, Sinval Pinto Brandão Filho Centro de Pesquisas Aggeu Magalhães 1 Department of Parasitology, School of Biomedical and Laboratory Sciences, College of Medicine and Health Sciences, University of Gondar, Gondar, Ethiopia. 2Department of zoology, Faculty of Life Science, Addis Ababa University, Addis Ababa, Ethiopia. 3 Aklilu Lemma Institute of Pathobiology, Addis Ababa University, Addis Ababa, Ethiopia. 4Department of Microbiology & Molecular Genetics, The Institute for Medical Research Israel-Canada, The Kuvin Centre for the Study of Infectious & Tropical Diseases, The Hebrew University – Hadassah Medical School, The Hebrew University of Jerusalem, 91120, Israel. 5Department of Microbiology, Immunology & Parasitology, Faculty of Medicine, Addis Ababa University, Addis Ababa, Ethiopia [email protected] Keywords: Population dynamics; Phlebotomus orientalis Abstract Background. Metema-Humera lowlands are endemic for kala-azar (visceral leishmaniasis). These lowlands are characterized by black clay soil and used for growing sesame, sorghum and cotton for commercial purposes. Objectives. The aim of this study was to study seasonal dynamics and habitat preference of Phlebotomus orientalis in extra-domestic habitats. Methods. CDC-light trap [CDCLT] and Sticky Trap [ST] were used to collect sand flies from various habitats before species identification by their morphological characteristics using appropriate keys. Data summarized and analyzed included: species, sex, density, habitats, type of trap used and date (month). Results. A total of 389,207 sand flies using CDC-LT (n=992) and ST (n=5730) were collected from May 17, 2011 to June 6, 2012. The highest Mean Monthly Density (MMD) of P. orientalis trapped by CDC-LT was found in thickets of Acacia seyal in March (MMD of 64.11 + 75.87). The corresponding highest MMD of P. orientalis trapped by STs was found in April (58.69 + 85.20) in agricultural field. No P. orientalis were caught in September using CDC traps and July October using sticky traps. The overall MMD of P. orientalis trapped by CDC-LT was 14. 83 + 28.03 in agricultural field, 19.37 + 36.42 in thickets of A. seyal, and 3.55 + 6.082 in dense mixed forest. Localitywise, the MMD of P. orientalis trapped by CDC-LT did not show significant statistical difference (p=0.135). Conclusions. Agricultural fields and thickets of A. seyal, habitats which exhibit extensive soil cracks and fissures, as opposed to dense mixed forests, serve as preferred breeding sites for P. orientalis. [email protected] Keywords: Sand flies, Phlebotominae, Lutzomyia longipalpis, Ecology Visceral leishmaniasis is a major public health issue in South America, where the disease is rapidly spreading. Changes in ecology and distribution of the principal vector, Lutzomyia longipalpis are among the factors accounting for the increasing incidence of the disease in this region. However, information about the ecology of L. longipalpis is still incipient, which may directly impair the implementation of effective control programs. Herein, the ecology of L. longipalpis was studied in a focus of visceral leishmaniasis in north-eastern Brazil. From August 2009 to August 2010, phlebotomine sand flies were monthly collected in four localities using CDC light traps (∼37 per month) and a lantern-baited Shannon trap with mouth aspirators. A total of 24,226 phlebotomine sand flies were collected with light traps and 375 with mouth aspirators. The most abundant species was L. longipalpis, representing 97.9% of the specimens collected with light traps and 91.5% with the mouth aspirator. Other species (Lutzomyia evandroi, Lutzomyia lenti and Lutzomyia sallesi) were found in low numbers. Most phlebotomine sand flies (94.6%) were collected at chicken coops and corrals. No significant correlation was found between the monthly abundance of phlebotomine sand flies and the monthly averages of temperature, relative humidity or rainfall. However, interestingly enough, 82.4% of L. longipalpis specimens were collected in months when relative humidity surpassed 75%. This study points out that this vector is well adapted to live in different habitats and to different climate conditions. It also suggests that some north-eastern populations of L. longipalpis may be more xerotolerant than southern populations. Further studies to assess the relationship between microclimate and L. longipalpis density in different Brazilian regions are advised. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 75 ID 31-P Ecology & Epidemiology ID 32-P Ecology & Epidemiology DIVERSITY OF PHLEBOTOMINE SAND FLIES IN AN AMERICAN CUTANEOUS LEISHAMANIASIS ENDEMIC AREA IN NORTH-EASTERN BRAZIL A REAL-TIME PCR FOR CHARACTERIZATION OF M U LT I P L E B L O O D S O U R C E S I N F I E L D COLLECTED PHLEBOTOMINE SAND FLIES Débora Elienai de Oliveira Miranda, Kamila Gaudêncio da Silva, Maria Aparecida da Gloria Faustino, Leucio Câmara Alves, Sinval Pinto Brandão-Filho, Filipe Dantas-Torres, Gílcia Aparecida de Carvalho Kamila Gaudêncio da Silva, Rayana Carla Silva de Morais, Débora Elienai de Oliveira Miranda, Abilene Cristina de Arruda Moura, Sinval Pinto Brandão-Filho, Milena de Paiva Cavalcanti, Filipe Dantas-Torres Department of Veterinary Medicine, Federal Rural University of Pernambuco, 52171900, Recife, Pernambuco, Brazil, Department de Immunology, Aggeu Magalhães Research Centre, 50670-420 Recife, Pernambuco, Brazil, Department of Veterinary Medicine, Università degli Studi di Bari, 70010 Valenzano, Bari, Italy, Academic Unit of Garanhuns, Federal Rural University of Pernambuco, Garanhuns, Pernambuco, Brazil, 55296-901. Departamento de Imunologia, Centro de Pesquisas Aggeu Magalhães, 50670-420 Recife, Pernambuco, Brazil, Dipartimento di Medicina Veterinaria, Università degli Studi di Bari, 70010 Valenzano, Bari, Italy. [email protected] [email protected] Keywords: Sand flies, source blood and qPCR Keywords: Ecology, phlebotomine and vectors Zoonotic leishmaniasis caused by Leishmania infantum is endemic in the international touristic Algarve region (AR) in southern Portugal being Phlebotomus perniciosus the only proven vector up to now. However, L. major DNA was recently detected in one Sergentomyia minuta specimen collected in this region. Nevertheless, and albeit up to now no autochthonous cutaneous leishmaniasis due to L. major or L. tropica has been detected in Portugal, their vectors (i.e. P. papatasi and P. sergenti, respectively) were also identified. We report a three years survey carried out to update the distribution, density, Leishmania infection rate and to identify blood-meal sources of phlebotomine sand flies in AR. From May to October 2011, 2012 and 2013, CDC light traps and sticky papers were set up in 11 sampling points during 3 consecutive days per month. Collecting places included domestic, peri-domestic and sylvatic environments. A total of 4961 sand flies (2569 males and 2392 females) were collected and identified morphologically. At least one specimen of the five known species present in Portugal (P. ariasi, P. papatasi, P. pernicious, P. sergenti and S. minuta) was captured. The predominant species was S. minuta followed by P. perniciosus highlighting the high plasticity of both species to adapt to different environments. In addition, blood meals revealed that P. perniciosus and S. minuta fed on a wide range of hosts (i.e. humans, horses, cattle, sheep, pigs, poultry, rabbits and lizards). Although Sergentomyia genus is known as a vector of Sauroleishmania sp. its epidemiological significance should not be neglected as they have already been suspected to be involved in L. donovani and L. major transmission among humans in India and Mali, respectively and L. infantum among dogs in Senegal. Our results reinforce the importance of systematic surveillance with extensive epidemiological surveys on Leishmania hosts and vectors to investigate the transmission, distribution and spreading of Leishmania sp. infection in order to detect areas of higher of risk of vector transmission and to implement specific control measures. Funding: This work was partially supported by EDENext (FP7INCO-CT-2010-261504). C. Maia (SFRH/BPD/44082/2008) holds a fellowship from Fundação para a Ciência e a Tecnologia, Ministério da Ciência, Tecnologia e Ensino Superior, Portugal. Phlebotomine sand flies are vectors of protozoa of the genus Leishmania, the causative agents of leishmaniasis. The ecology of phlebotomine sand flies is of research interest because it can provide background information for a better understanding of the epidemiology of leishmaniasis and to predict the spatial and temporal variations in the risk of infection in a given area. American cutaneous leishmaniasis (ACL) is among the most important tropical diseases in Brazil and in several developing countries in the Neotropics. The present study aimed to identify the different species that make up the phlebotomine sand flies fauna in an area of the municipality of the Ipojuca, Pernambuco state, Brazil. Phlebotomine sand fly collections were conducted from August 2013 to February 2014, using 7 CDC light traps for three consecutive nights from 17:00 pm to 6:00 am, on a monthly basis. Each trap was placed at 1.5 m above the ground, being installed in three different environments: indoors, peridomicile (i.e., forest edge) and in the forest. The capture sites were usually near the burrows of wild animals, trunks of large trees, in shady and moisturerich environments. The captured specimens were examined, separated according to sex and kept in 70% ethanol. The sandflies were mounted on slides and identified morphologically. A total of 2,889 specimens belonging to 17 species were identified, namely: L. choti (n = 1,680), L. whitmani (n = 495), L. quinquefer (n = 223), L. sordellii (n = 209), L. longispina (n = 84), L. amazonensis (n = 52), L. evandroi (n = 49), L. brasiliensis (n = 28), L. naftalekatzi (n = 27), L. barrettoi barrettoi (n = 17), L. capixaba (n = 10), L. shannoni (n = 4), L. aragaoi (n = 2) L. complexa (n = 5) L. furcata (n = 2), L. wellcomei (n = 1) and L. ayrozai (n = 1). Males (n = 1,431) predominated over females (n = 1,458). L. choti and L. whitmani were the most abundant species, comprising together 75.2% of the specimens collected. The study demonstrates an important variety of phlebotomine sand fly species in the studied area and that some of them may be associated with human cases of ACL. The finding of L. whitmani, a proven vector of Leishmania braziliensis in several Brazilian regions indicates its probable involvement in the transmission of ACL in this municipality. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 76 ID 33-P Ecology & Epidemiology ID 34-P Ecology & Epidemiology A THREE CONSECUTIVE YEARS PHLEBOTOMINE SAND FLY SURVEY IN A ZOONOTIC LEISHMANIASIS ENDEMIC FOCUS, THE ALGARVE REGION, PORTUGAL NOCTURNAL ACTIVITIES AND HOST PREFERENCES OF P. ORIENTALIS IN EXTRADOMESTIC HABITATS OF KALA-AZAR ENDEMIC METEMA–HUMERA LOWLANDS, NORTH WEST ETHIOPIA C.Maia1,2,3, J. Cristóvão 1,2, F. B. Freitas1, M.O. Afonso1,4, L. Campino1,5 1 Unidade de Parasitologia Médica, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Lisboa, Portugal 2Centro de Malária e outras Doenças Tropicais 3Faculdade de Medicina Veterinária, Universidade Lusófona de Humanidades e Tecnologias, Lisboa, Portugal 4Unidade de Parasitologia e Microbiologia Médicas 5Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, Faro, Portugal [email protected] Keywords: Phlebotomine sand fly survey; zoonotic leishmaniasis; Algarve; Portugal Zoonotic leishmaniasis caused by Leishmania infantum is endemic in the international touristic Algarve region (AR) in southern Portugal being Phlebotomus perniciosus the only proven vector up to now. However, L. major DNA was recently detected in one Sergentomyia minuta specimen collected in this region. Nevertheless, and albeit up to now no autochthonous cutaneous leishmaniasis due to L. major or L. tropica has been detected in Portugal, their vectors (i.e. P. papatasi and P. sergenti, respectively) were also identified. We report a three years survey carried out to update the distribution, density, Leishmania infection rate and to identify blood-meal sources of phlebotomine sand flies in AR. From May to October 2011, 2012 and 2013, CDC light traps and sticky papers were set up in 11 sampling points during 3 consecutive days per month. Collecting places included domestic, peri-domestic and sylvatic environments. A total of 4961 sand flies (2569 males and 2392 females) were collected and identified morphologically. At least one specimen of the five known species present in Portugal (P. ariasi, P. papatasi, P. pernicious, P. sergenti and S. minuta) was captured. The predominant species was S. minuta followed by P. perniciosus highlighting the high plasticity of both species to adapt to different environments. In addition, blood meals revealed that P. perniciosus and S. minuta fed on a wide range of hosts (i.e. humans, horses, cattle, sheep, pigs, poultry, rabbits and lizards). Although Sergentomyia genus is known as a vector of Sauroleishmania sp. its epidemiological significance should not be neglected as they have already been suspected to be involved in L. donovani and L. major transmission among humans in India and Mali, respectively and L. infantum among dogs in Senegal. Our results reinforce the importance of systematic surveillance with extensive epidemiological surveys on Leishmania hosts and vectors to investigate the transmission, distribution and spreading of Leishmania sp. infection in order to detect areas of higher of risk of vector transmission and to implement specific control measures. Funding: This work was partially supported by EDENext (FP7-INCO-CT2010-261504). C. Maia (SFRH/BPD/44082/2008) holds a fellowship from Fundação para a Ciência e a Tecnologia, Ministério da Ciência, Tecnologia e Ensino Superior, Portugal. Wossenseged Lemma1, Habte Tekie2 , Ibrahim Abassi3, Meshesha Balkew3, Teshome Gebre-Michael3, Alon Warburg3 ,Asrat Hailu5 1 *Department of Parasitology, School of Biomedical and Laboratory Sciences, College of Medicine and Health Sciences, University of Gondar, Gondar, Ethiopia. 2 Department of zoology, Faculty of Life Science, Addis Ababa University, Addis Ababa, Ethiopia. 3Department of Microbiology & Molecular Genetics, The Institute for Medical Research Israel-Canada, The Kuvin Centre for the Study of Infectious & Tropical Diseases, The Hebrew University – Hadassah Medical School, The Hebrew University of Jerusalem, 91120, Israel. 3Aklilu Lemma Institute of Pathobiology, Addis Ababa University, Addis Ababa, Ethiopia. 5Department of Microbiology, Immunology & Parasitology, Faculty of Medicine, Addis Ababa University, Addis Ababa, Ethiopia [email protected] Keywords: Nocturnal Activities, Phlebotomus orientalis Background: P. orientalis takes blood from a variety of wild and domestic animals and transmit L. donovani from unknown reservoir hosts to human in extra-domestic habitats in Metema - Humera lowlands. Objective: The aim of this study was to determine the nocturnal activities of P. orientalis and their preferred host for blood meal during night time. Methods: Hourly collections of P. orientalis using CDC traps from farm, near goat and sheep night shelter and thickets of Acacia seyal were used for the study nocturnal activities of P. orientalis. Host preference study of P. orientalis towards rodents was studied using Turner's box trap. Self bait technique was used to study the host preference of P. orientalis towards human in addition to blood meal analysis of this vector using Reverse Line Blot (RLB) of cytochrome b PCR product. Result. A total of 6083 P. orientalis (2702 male and 3381 female) were collected at one hour interval from farms near camps, sparse thickets of Acacia seyal and near animal shelters (sheep and goat) using 22 CDC traps during January, February March and May, 2013. Nocturnal activities of female P. oriental were different at different hours of the night in the farm (p=0.023) and near animal shelter (p=0.00). However, the activities were the same for the different hours of the night in the thickets of Acacia seyal (p=0.144). The peaks activities of P. oriental were at 1.00 a. m near animal shelter (134.0 ±7.21) compared to 3.00 a. m (66.33 ± 46.40) in the farm. A total of 984 female P. orientalis from farm near the camp and animal house were collected at one hour intervals using self bait. The highest mean landing of P. orientalis were 38.50 ± 31.88 at 12 pm with maximum number of 86 with no statistical difference for mean number of P. orientalis biting man (p = 0.066). P. orientalis were attracted more to human than sheep and goats ((p=0.000). P. orientalis were not attracted to the different species of rodents. Blood meal analysis of the 25 blood fed P. orientalis using cyt b PCR and RLB captured 7 human(28%), 9 mixed (human and cattle) (36%), 2 cattle(8%). There were samples (28%) with bands on agarose gel but not in nitrocellulose membrane indicating the existence of other animals outside the 11 probes used. Conclusion: P. orientalis feed on goats, sheep, cattle and human throughout the night with peak biting after mid night. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 77 ID 35-P Ecology & Epidemiology ID 36-P Ecology & Epidemiology PREVALENCE OF LEISHMANIA SPP. INFECTION AND HOST - BLOOD FEEDING OF SANDFLIES TRANSMITTERS OF CUTANEOUS LEISHMANIASIS IN AN ENDEMIC LOCALITY IN THE DISTRICT OF CAPIRA, PANAMA A STUDY ON NOCTURNAL ACTIVITY OF SAND FLIES IN A LEISHMANIASIS ENDEMIC VILLAGE LOCATED IN AYDIN PROVINCE OF TURKEY Suha K. Arserim1, Mehmet Karakus2, Yusuf Ozbel3 Chystrie Rigg1,2, Anayansi Valderrama3, Chavez4 y Azael Saldaña1 José E. Calzada1, Luis F. 1 Celal Bayar University, Vocational School of Health Services, Manisa, TURKEY; 2Ege University Institute of Health Sciences, Bornova, Izmir, TURKEY; 3Ege University Faculty of Medicine, Department of Parasitology, Bornova, Izmir, Turkey 1 Department of Parasitology, Gorgas Memorial Institute for Health Studies, Panama; Central American Program of Masters in Entomology, University of Panama, Panama; 3Department of Medical Entomology, Gorgas Memorial Institute for Health Studies, Panama; 4Institute of Tropical Medicine (NEKKEN), Nagasaki University, Japan. 2 [email protected] Keywords: Sand fly, nocturnal activity, Turkey [email protected] Keywords: Leishmaniasis, infection, Blood Feeding, Panama. The community of Trinidad de las Minas in the District of Capira, Province of Panama, is located in a region where an intense transmission of cutaneous leishmaniasis (CL) have been reported. In this area, the increase of CL cases ranges between 137 to 508 per year, with an incidence that corresponds to 20% of the whole country. The purpose of this work is to demonstrate in this community, the prevalence of Leishmania spp. infection and host-blood feeding in anthropophilic and zoophilic sandflies, before and after vector control intervention with insecticide thermal fogging (ITF). From April 2010 to June 2011, sandflies where collected using CDC light traps, inside (domicile) and around (peridomicile) of 24 houses. To evaluate the Leishmania infection, we analyzed the DNA sand fly pools (main species, 5-10 individuals) by ITS-1 gene PCR/sequencing. Positive pools were analyzed by cytochrome b PCR/sequencing, to differentiate the host-blood feeding patterns (domestic, wild mammals or birds). Of 455 analyzed pools, 253 resulted positives for PCR-ITS1. Of them, 164 correspond to important leishmaniasis vectors found in this community: Lutzomyia trapidoi, Lu. gomezi and Lu. panamensis. The estimated prevalence to 95% depending on the algorithms of the epitool tool (pooled prevalence for variable pool size and perfect tests) was 0.09 to 0.11 before and after the intervention respectively. In the detection of host-blood feeding, it was found that birds and pigs were the main sources of blood, followed by humans and dogs. These findings demonstrate the dynamics of Leishmania infection and blood feeding in vector sandflies, before and after an ITF, a method often used to control mosquitoes and other arthropods in Panama. The results confirm the infection of the main species of anthropophilic sandflies with Leishmania spp. in this hyperendemic focus of CL. A close association between these vector species, humans and some domestic animals was also supported, emphasizing the importance of implementing active surveillance using these parameters to predict outbreaks and develop new control strategies or integrate existing ones to reduce cutaneous leishmaniasis in this region. The nocturnal activity of sand flies in an endemic focus for canine and cutaneous leishmaniasis in western part of Turkey was studied using CDC light traps. The traps were set up in different micro-localities, such as human house, cattle shed, chicken coop and sheep area within 60-70 meters in diameter. In the study night, a total of 263 sand flies, six species belonging to Phlebotomus and two species belonging to Sergentomyia genera, were caught. P. tobbi was found to be abundant species (60.46%) while P. neglectus/syriacus was second one (21.29%). The activity of sand flies started at 21:00-22:00h, reached maximum between 01:00 and 03:00h and steadily decreased thereafter and finished at 05:00h in all micro-localities. Female sand flies was always 2-3 times in high number than males. The study was showed the highest activity of probable vector species, P. tobbi, in the area and proved the importance of the studies on seasonal and daily activity of sand flies in endemic areas. This study was partially funded by EU grant GOCE-2003-010284 EDENext. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 78 ID 37-P Ecology & Epidemiology ID 38-P Ecology & Epidemiology SEASONAL ACTIVITIES AND ECOLOGICAL F E AT U R E S O F S A N D F L I E S ( D I P T E R A : PSYCHODIDAE: PHLEBOTOMINAE) IN AYDIN MOUNTAINS AND SURROUNDINGS LOCATED IN W E S T E R N T U R K E Y, E N D E M I C A R E A F O R CUTANEOUS LEISHMANIASIS BIONOMY OF PHLEBOTOMINE SANDFLIES (PHLEBOTOMINAE) IN AN OUTBREAK AREA OF VISCERAL LEISHMANIASIS IN CORRIENTES, ARGENTINA Araujo AV 1, Benítez Ibalo AP 1, Rosa JR 2; Oscherov EB 1; Salomón OD 3 Suha K. Arserim1, Ahmet Mermer2, I. Cuneyt Balcioglu3, V. Ozge Ermis4; Yusuf Ozbel5 1 Celal Bayar University Vocational School of Health Services, Manisa, Turkey; 2Ege University, Science Faculty, Department of Zoology, Bornova, Izmir, Turkey; 3Celal Bayar University Faculty of Medicine, Department of Parasitology, Manisa, Turkey; 4 Celal Bayar University Institute of Science, Manisa, Turkey; 5Ege University Faculty of Medicine, Department of Parasitology, Izmir, Turkey 1 Lab. Biología de los Parásitos. Facultad de Ciencias Exactas y Naturales y Agrimensura. Universidad Nacional del Nordeste (UNNE). Red de Investigación de la Leishmaniasis en Argentina (REDILA). 2 Lab. Entomología. Instituto de Medicina Regional. UNNE. REDILA. 3 Instituto de Medicina Tropical (InMET), Puerto Iguazú, Misiones, Argentina. REDILA. [email protected] [email protected] Keywords: Leishmania , Epidemiology, Lutzomyia Keywords: Sand fly, Seasonal activity, Leishmaniasis, Aydın Mountains The abundance study in time and space of Phlebotominae through the area of the City of Corrientes (27°29′S;58°49′W) was based in human and canine cases with Visceral Leishmaniasis (VL) reported in the province and the previous records of the urban vector Lutzomyia longipalpis in the city. This preliminary study involves collection performed in three sites according to its transitional environments: periurban-rural (ENU), periurban-urban (EPU) and urban-centric (EU). In each environment three households were selected based on the ´worst scenario´ criteria. From March to December 2013 the captures were made fortnightly during three consecutive days using CDC type-light trap in each peridomiciliary habitat, from 17:00hs to 8:00hs. In total 449 Phlebotominae were captured: Lu. longipalpis (51.2%), Nyssomyia neivai (43.7%), Migonemyia migonei (2.2%), Micropygomia quinquefer (2.0%) and Evandromyia females from cortelezzii complex (0.9%). The sandfly maximal abundance was in March (32.7%) with an average temperature of 22ºC, and minimum abundance during August (0.2%) with temperatures of approximately 14,3ºC. Lu. longipalpis was recorded in all the seasons with the highest abundance in March and April (n=119) during autumn, and the lowest abundance (n=27) in winter, with a progressive increase of the abundance in the captures from September, at the beginning of the spring. The main abundance of Ny. neivai was also in autumn, but its captures increased since October. Although Lu. longipalpis was recorded in the three environments, was more abundant in EPU and EU, while Ny. neivai was more abundant in ENU. Mi. migonei was recorded in autumn and spring in ENU and EPU but due to the low number collected may be considered as an occasional specie. The consistent presence of Lu. longipalpis in time and space in the urban area and the occasional captures of Mi. migonei, main and putative vectors of Leishmania infantum respectively, besides the reports of autochthonous cases of human and canine VL, suggests a risk of transmission along the year. However, this risk is highly heterogeneous in time and space, lower in winter despite of the fact that the actual incidence of cases could increase early in this season due to the abundance vectors peaked in autumn and the parasite intrinsic incubation time. Financial support: Proyecto (F008/09) Res. Nº 1080/09. Periodo 2010-2013. UNNE. PICTO-ANLIS 2011. The aim of this study was to determine (a) the seasonal activities and ecological features of sand fly population, (b) identifying sand fly fauna, (c) preparing maps for showing the distribution of existing species, and (d) analyzing the diversity and population sizes of sand fly species in The Aydın Mountains and surroundings located in western Turkey, an important endemic focus for cutaneous and canine leishmaniasis. During the entomological survey 10 locations were selected according to the altitude and sand flies were collected using CDC light traps and sticky paper traps from June 2009 to July 2010 by 15-day intervals in summer season and 30-day intervals in winter season. Temperature, humidity and general environmental parameters of study area were recorded using data loggers. Totally, 6817 sand flies were collected and dissected. The identification was made according to the Mediterranean keys. In total, 12 species of sand flies were identified, which are 9 species belonging to genus Phlebotomus (P. papatasi 5.79%, P. similis 4.44%, P. alexandri 4.20%, P. mascittii 0.28%, P. neglectus/syriacus 29.91%, P. burneyi 0.62%, P. tobbi 22.58%, P. simici 3.45% and P. brevis 0.44%) and 3 species are belonging to genus Sergentomyia (S. dentata 22.81%, S. antennata 1.55% and S. minuta 2.39%). In conclusion, 12 sand fly species are detected in the study area and maps were produced. P. neglectus, P. tobbi and P. similis, the well known vector species for Leishmania infantum and L. tropica were found dominant species by several analyses, respectively. The activity period was determined as between April and October, for 7 months. Population abundance of P. similis was the highest at the altitude of 400-600m in July while P. neglectus and P. tobbi were found more abundant at the altitude of 200-400m in September. These results on sand fly ecology area could be useful for establishing or improving effective control programs. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 79 ID 39-P Ecology & Epidemiology ID 40-P Ecology & Epidemiology ATTRACTION OF ETHIOPIAN PHLEBOTOMINE SAND FLIES (DIPTERA: PSYCHODIDAE) TO LIGHT AND SUGAR-YEAST MIXTURES (CO2) ECOLOGICAL NICHE MODELING PREDICTS S O U T H WA R D E X PA N S I O N O F L U T Z O M Y I A F L AV I S C U T E L L ATA , V E C TO R O F D I F F U S E CUTANEOUS LEISHMANIASIS IN SOUTH AMERICA, UNDER CLIMATE CHANGE Oscar David Kirstein, Roy Faiman, Araya Gebreselassie, Asrat Hailu, Teshome Gebre-Michael and Alon Warburg Department of Microbiology and Molecular Genetics, The Institute for Medical Research Israel-Canada, The Kuvin Centre for the Study of Infectious and Tropical Diseases, The Hebrew University – Hadassah Medical School, The Hebrew University of Jerusalem, Israel. [email protected] Keywords: Attractants, Phlebotomus orientalis, Visceral leishmaniasis, Yeast fermentation. Visceral leishmaniasis (VL) known as Kala-Azar is a serious systemic disease caused by Leishmania donovani parasites (Trypanosomatidae: Kinetoplastida). The disease is prevalent in the Indian Sub-continent, East Africa and Brazil. In Africa, the worst affected regions are in Sudan, with an estimated 15,000-20,000 cases annually and, Ethiopia with 5,000-7,000 cases a year. The main vector of VL in Sudan and Northern Ethiopia is Phlebotomus orientalis, a sand fly frequently found in association with Acacia spp and Balanite spp woodlands. Methods: To optimize sampling of sand flies for epidemiological studies in remote areas we tested different means of attraction. Miniature suction traps employing 2AA batteries (3 V) were deployed in the up-draft orientation and baited with chemical light-sticks (Red, Yellow and Green), or bakers' yeast in sugar solution (emitting CO2 and perhaps other attractants). These traps were compared with standard CDC incandescent light traps employing 6 V 20 batteries. Trials were conducted during two consecutive years at different localities around Sheraro, a town in West Tigray, Northern Ethiopia.Results: The sand fly species composition was similar but not identical in the different locations tested with different Sergentomyia spp. predominating. Phlebotomus spp. comprised less than 1% of the catches during the first year trials (November – December 2011) but increased markedly during the second year trials performed later in the dry season at the height of the sand fly season in February 2012. Although there did not appear to be a species bias towards different light wave-lengths, fermenting yeast in sugar solution attracted relatively more Phlebotomus spp. and Sergentomyia schwetzi.Conclusions: Although the standard 6 V CDC incandescent light traps captured more sand flies, light-weight (~350 g) 3 V suction traps baited with chemical light-sticks were shown to be effective means of monitoring sand flies. Such traps operated without light and baited with yeast in sugar solution caught relatively more Phlebotomus spp. Bruno Moreira de Carvalho1,2,3, Mariana Moncassim Vale2, Maria Lucia Lorini4, Margarete Martins dos Santos Afonso1, Elizabeth Ferreira Rangel3 1 Programa de Pós-Graduação em Ecologia e Evolução, Universidade do Estado do Rio de Janeiro; 2Departamento de Ecologia, Instituto de Biologia, Universidade Federal do Rio de Janeiro; 3Laboratório de Transmissores de Leishmanioses, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz; 4Departamento de Ciências Naturais, Instituto de Biociências, Universidade Federal do Estado do Rio de Janeiro. [email protected] Keywords: Ecological niche modeling, climate change, Lutzomyia flaviscutellata According to the Fifth Assessment Report of the Intergovernmental Panel on Climate Change, climate change will exacerbate health problems that already exist. Leishmaniasis is a climate-sensitive disease, occupying a characteristic climate space that is strongly affected by changes in rainfall and temperature. Climate change, therefore, should alter the distribution of leishmaniasis vectors. We modeled the climatic suitability of Lutzomyia flaviscutellata, main vector of diffuse cutaneous leishmaniasis (DCL) in South America, and projected the models under climate change scenarios to assess possible range changes. To provide occurrence data, a database of 328 records of L. flaviscutellata was compiled from the literature, museum collections and Brazilian Health Departments. Six bioclimatic variables derived from temperature and precipitation were used as predictors. Models were produced based on eight different algorithms – Bioclim, Domain, Mahalanobis Distance, Generalized Linear Model, Generalized Additive Model, Maxent, Random Forest and Support Vector Machines – and evaluated by a threshold-independent method (AUC). Ensemble models were produced by overlapping binary predictions of each algorithm. Future projections used 15 most recent climate models for 2050 (CMIP5), in climate change scenarios representing two greenhouse gas concentration pathways: stabilization (RCP4.5) and high increase (RCP8.5). Single-models produced by each algorithm and climate model showed fair to good performance (0.71<AUC<0.87), but resulted in quite dissimilar predictions. Compiled records of Lutzomyia flaviscutellata are located in Bolivia, Brazil, Colombia, Ecuador, French Guiana, Peru, Suriname, Trinidad and Tobago and Venezuela. Currently, the species is widely distributed in the Amazon, with some records in Atlantic Forest and savannah regions to the south. Most consensus models agree that the area of climatic suitability of L. flaviscutellata should expand in both climate change scenarios, and all consensus models agree that the vector should experience improved climatic suitability towards the south. Human cases of DCL in southeast Brazil are currently rare, although the disease seems to be gradually expanding its occurrence southward. If the vector reaches these climatically suitable areas and its dispersion is followed by competent hosts and parasites, these can become DCL risk areas, especially because these are the most populated areas within the species' range. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 41-P Ecology & Epidemiology ID 42-P Ecology & Epidemiology IDENTIFICATION OF PLANTS FED UPON BY PHLEBOTOMINE SAND FLIES IN ETHIOPIA H A B I TAT S U I TA B I L I T Y F O R L U T Z O M Y I A (NYSSOMYIA) WHITMANI AND AMERICAN CUTANEOUS LEISHMANIASIS (ACL) IN BRAZIL Oscar David Kirstein; Ibrahim Abassi; Araya Gebreselassie; Teshome Gebre-Michael; Asrat Hailu and Alon Warburg Department of Microbiology and Molecular Genetics, The Institute for Medical Research Israel-Canada, The Kuvin Centre for the Study of Infectious and Tropical Diseases, The Hebrew University – Hadassah Medical School, The Hebrew University of Jerusalem, Jerusalem - Israel [email protected] Keywords: PCR, Plant DNA, Sand fly, Ethiopia Only the female sand flies transmit Leishmania parasites when they suck blood required for maturation of the eggs. However, both sexes feed on plant-derived sugars by imbibing plant sap, nectar and/or honeydew. Therefore, determining the sources of sand fly sugar meals can improve our understanding of the ecology and transmission of leishmaniasis. The attraction of sand flies to certain habitats probably depends on the vegetation and their preference for certain plants as food sources, resting places or even breeding habitats. Thus, certain plant communities may comprise risk factors for contracting leishmaniasis. For example, the sand fly Phlebotomus orientalis, vector of visceral leishmaniasis (VL) in Sudan and Northern Ethiopia is frequently associated with wooded areas dominated by Acacia sp and Balanites sp. In fact, Acacia forests are commonly perceived as an important factor in contracting Kala Azar in these areas. A botanical survey was conducted in different areas around Sheraro town, in Tigray, northern Ethiopia. Twenty four of the plant species predominating during the dry season were collected and identified based on classical and molecular plant taxonomy. For molecular identification, PCR systems were developed for three chloroplast genes rbcl, MatK, ATP synthase and the nuclear ribosomal gene (ITS1). Plant specific primers were designed after DNA alignment of DNA sequences and primer annealing sites were selected in the conserved sequences. The PCR amplified regions were sufficiently variable to identify the different species by DNA sequencing. The two primer pairs for rbcl and ATP synthase were shown to be the most suitable. In preliminary experiments using the new PCR system, we successfully amplified plant DNA from Ph. orientalis sand flies collected in different ecotopes. Further studies are ongoing to adapt a rapid and inexpensive DNA identification protocol using reverse blotting in order to replace the more costly time-consuming DNA sequencing. 80 Simone M. da Costaa, José L. P. Cordeirob, Margarete M.S. Afonsoa, Elizabeth F. Rangela Laboratório de Transmissores de Leishmanioses, IOC/FIOCRUZ, Av. Brasil n° 4365, 21040360, RJ, Brazil; bCampus Fiocruz da Mata Atlântica, CFMA/Presidência, FIOCRUZ, RJ, Brazil. [email protected] Keywords: Lutzomyia (Nyssomyia) whitmani, Leishmaniose Tegumentar Americana, alterações ambientais The impact of environmental change as well as those determined by human activity, combined with different aspects of the vector, parasite, and reservoir host ecology, play an important role in the dissemination of ACL in Brazil. In this context highlight L. (N.) whitmani, most important vector of L. (V.) braziliensis and L. (V.) shawi. The characterization of factors that influence the spatial distribution of sand fly species is a first-step for understanding the ecology of ACL. Environmental suitability for ACL and L. (N.) whitmani for Brazil was evaluated through ENFA. For data related to the occurrence of the disease, municipal localities with records of endemic areas for ACL (N = 1882) were used. For occurrence of phlebotomines the municipalities with confirmed record of vector (N = 633) were considered in L. (N.) whitmani Model. The models were developed using 14 Environmental Variables (EV) (12 bioclimatic, altitude and NDVI).Cross-validation of the models' quality resulted in a Boyce index of 0.89 ± 0.10 and 0.84 ± 0.16, for the ACL Model and the L. (N.) whitmani Model respectively, indicating a satisfactory predictive capacity of both models. L. (N.) whitmani was identified by the model as a species that occurs in a particular condition in the study area, in the context of the average condition of all EVs, and suggests that the species is moderately tolerant of a range of environmental conditions (M = 0.87, T = 0.62). Many evidences indicate that this sand fly has capacity to survive environmental changes and to adapt to new ecological niches, tolerating effects of climatic alterations. Analysis by EVs showed that L. (N.) whitmani "prefers" (more suitable habitat) and is more frequent in regions with relative less rainfall and warm temperatures. These characteristics are in accordance with previous analysis discussing the distribution of this sand fly vector in Brazilian biomes, occurring in high frequency in Cerrado and Atlantic Rainforest, with greater amplitude in these Biomes. The ACL Model indicate that more suitable areas for ACL moderately differs from the average conditions across the entire distribution area, suggesting that ACL is not too selective of its environment, and is tolerant of a range of environmental conditions (M=0.53; T=0.77). The ACT Model identifies more suitable areas related to intermediate density vegetation (in NDVI EV) and zones with annual precipitation around 1500 mm. Financial support: LTL/IOC; Fiocruz/Faperj; Capes. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 43-P Ecology & Epidemiology ID 44-P Ecology & Epidemiology A FIELD STUDY TO EVALUATE THE PERFORMANCE OF TWO MODIFIED CDC LIGHT TRAPS FOR COLLECTING SANDFLIES COMPARED WITH THE TRADITIONAL CDC TRAP LIGHT POPULATION COMPOSITION AND DYNAMICS OF L U T Z O M Y I A L O N G I PA L P I S A N D O T H E R PHLEBOTOMINAE SPECIES IN CLORINDA, NORTHERN ARGENTINA Gould, Ignacio Tomás1, Fernández María Soledad2,3, Pérez Adriana Alicia4, Salomón Oscar Daniel2,3 Andrea Gómez Bravo1, Ignacio Gould2,4, Marcelo Abril1, Oscar Daniel Salomón3,4 Redila 1Centro Nacional de Diagnóstico e Investigaciones en Endemoepidemias; Instituto Nacional de Medicina Tropical; 3Consejo Nacional de Investigaciones Científicas y Técnicas; 4Departamneto de Ecología, Genética y Evolución, FCEN, UBA . Fundación Mundo Sano, Buenos Aires, Argentina; 2. Centro Nacional de Diagnóstico e Investigaciones Endemo-epidémicas, Buenos Aires, Argentina; 3. Instituto Nacional de Medicina Tropical, Puerto Iguazú, Misiones, Argentina; 4. Consejo Nacional de Investigaciones Científicas y Técnicas, CONICET Argentina. Red de Investigación de Leishmaniasis en Argentina, REDILA 2 [email protected] 81 1 [email protected] Keywords: Sanflies, Light Traps, Lutzomyia longipalpis, Nyssomyia whitmani Keywords: Lutzomyia longipalpis, Visceral leishmaniasis, Phlebotominae. In recent years numerous surveys have been conducted in the Argentina to study the distribution and dispersal of sandflies due to an increment in the number of cases of visceral and tegumentary leishmaniasis. The aim of this work was to compare two local developed types of light traps for sandflies with the traditional CDC trap light (CDC). Both news traps are a modified CDC trap light with a ring of leds, one of white light (WL) and the other of black light (BL). Traps were compared according to the occurrence and the number of individuals of sandflies, Lutzomyia longipalpis, the main Leishmania infantum vector and Nyssomyia whitmani, a L. braziliensis vector. In order to test the traps 21 sample points were choosing in forest, rural and urban scenarios in the city of Puerto Iguazú and around it. A Latin Square design was used for the evaluation of the type of trap. At each site, the three types of traps were randomly assigned during three consecutive nights. The variation in the occurrence and numbers of individuals by species, according to the type of trap, the sex and their interaction was evaluated using Generalized Linear Mixed Models. For Lu. longipalpis, 300 individuals captured in 9 sites (urban and rural environmental) and for Ny. whitmani 1443 individuals captured in 11 sites (urban, rural and forest environmental) were included in the analysis. For both species, no differences were found for the occurrence between the three types of trap for both sex (p>0.05 in all cases). For Lu. longipalpis, for both sexes, the number of captured individuals didn´t differ between BL and CDC trap, neither between CDC and WL trap, but was higher for BL compared to WL trap (p=0.022). For Ny. whitmani we found no differences between types of trap for females (p>0.05), but for males we found that the traditional CDC and the WL traps captured more individuals that the BL traps (p<0.015 ). These results suggest that the new types of trap could be used as an alternative to the traditional CDC trap to detect the occurrence of these species in surveillance studies. In relation to the estimation of abundance by species and sex, these results shows the importance of using the same type of trap or combination of traps to compare species abundances, to avoid biases due to the methodology. Regarding the design, while a small number of sites were sampling, it covered a wide range of environments. In Argentina, the first record of Lu. longipalpis in an urban scenario was reported in Clorinda city (25°17′29″S 57°43′06″W) during 2004, locality on the border with Paraguay close to Asunción city that already has visceral leishmaniasis transmission. The local health authorities of Clorinda reported cases of canine visceral leishmaniasis afterward the first vector record. During 2010 a new entomological survey showed the persistence of Lu. longipalpis in critical sites of Clorinda. Aimed at describing the spatial and temporal distribution of Phlebotominae abundance in Clorinda, 20 sites were sampled on two consecutive nights, from January to December 2012, using CDC-like light minitraps. The selected sites were peridomestic habitats in the urban area, based on the Network for Leishmaniasis in Argentina (REDILA) criteria on 'worst scenario', and entomoepidemiological background from previous works. The Phlebotominae collected were separated locally and identified at a REDILA laboratory. Minimum and maximum temperature values during sampling days were recorded through a local meteorological station (CLORICOOP). From January to December 2012, the species captured were: Lu. longipalpis, Migonemyia migonei, Nyssomyia whitmani, and Bruptomyia sp. The most abundant species was Lu. longipalpis 98.73% (4740/4801), followed by Bruptomyia sp 0.62% (30/4801), Mg. migonei 0.42% (20/4801); and Ny. whitmani 0.23% (11/4801). Lu. longipalpis was present in all the sampling sites during the whole year. The month showing the greatest number of individuals was March (beginning of autumn) with 703 individuals/month, this month being the one when Lu. longipalpis was most abundant. July (winter) was the month with lower abundance with a total of 12 individuals, represented by Lu. longipalpis. During 2012 the average temperatures in Clorinda ranged between 29.70°C maximum (summer) and 19.39°C minimum (winter), and the average relative humidity was 79.27%. Lu. longipalpis turned out to be the species with the highest prevalence in all the urban area, but with spatial and temporal heterogeneous abundance due to micro-habitat characteristics and seasonal and daily climate variations, respectively. Identification of the sites and moments with the greatest abundance of Lu. longipalpis will result in a contribution to guide the control measures of this species. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 45-P Ecology & Epidemiology ID 46-P Ecology & Epidemiology DESCRIPTION OF POPULATION DYNAMICS AND HOURLY ACTIVITY OF LUTZOMYIA LONGIPALPIS IN THE CITY OF POSADAS, ARGENTINA COMPARISON OF CDC LIGHT AND GRAVID TRAPS FOR THE COLLECTION OF SANDFLY FAUNA IN BHUBANESWAR, ORISSA (INDIA) Soraya Alejandra Acardi1, Iris Ingrid Steinhorst2, Maria Soledad Santini3, Adolfo Enrique Sandoval2, Oscar Daniel Salomon4 Karina Mondragon-Shem1, 2, Melissa Bell1, Mrutyunjay Suar3, Parimita Routray1, Laura Posada4, Mary Cameron 1 1 1 London School of Hygiene and Tropical Medicine, England, UK | 2 Liverpool School of Tropical Medicine, England, UK | 3 Kalinga Institute of Industrial Technology –KIIT, Bhubaneswar, Orissa, India | 4 Programa de Estudio y Control de Enfermedades Tropicales PECET, Universidad de Antioquia, Colombia. Laboratorio de Biología Molecular Aplicada (LaBiMAp - UNaM). Misiones, Argentina. 2 Instituto Municipal de Vigilancia y Control de Vectores, Secretaria de Calidad de Vida, Municipalidad de Posadas. 3 Centro Nacional de Diagnostico e Investigación en Endemo-Epidemias (CeNDIE) Buenos Aires, Argentina. 4 Instituto Nacional de Medicina Tropical (INMeT) Puerto Iguazú, Misiones, Argentina. 82 [email protected] [email protected] Keywords: Lutzomyia longipalpis, population dynamics, hourly activity Lutzomyia longipalpis is the vector of Leishmania infantum in the city of Posadas, the first urban visceral leishmaniasis focus in Argentina. To describe its population seasonal dynamics and night activity phlebotomine sandflies were captured with CDC mini light traps placed overnight at two sites, two consecutive nights monthly from February 2009 to March 2010. Then the correlation between average monthly abundance and temperature, relative humidity (maximum, minimum and average for both) and rainfall for each month, 30 and 60 days before sampling (DBS) was analyzed by Spearman correlations (p value <0.05). In three houses with ≤ 30 sandflies a multi-sampling adapter for CDC mini light traps was placed two consecutive nights at each site from February to April of 2011, and it was programmed to change every two hours from 17:00 to 07:00hs. The association of trap/night average abundance with average temperature (T) and relative humidity (RH) of each period were studied by Spearman correlations. Paired t tests were performed to assess differences between abundance before and after 23:00-1:00hs period (corrected p value <0.008). Species were identified according to Galati (2003). The longitudinal study captured 698 Lu. longipalpis (male: female ratio 5.49:1), a bimodal pattern was observed with two peaks of activity, one in March (end of summer) and another in December (end of spring). Sandfly abundance showed a positive correlation with maximum, minimum and average temperature for the month of sampling and 30 DBS and with maximum temperature 60 DBS. In hourly activity study 250 Lu. longipalpis were captured (male: female ratio 5.75:1), the abundance had two peaks of activity at 19:0021:00hs and 1:00- 3:00hs. The least activity was from 23:00 -1:00hs for all months. Males are present during all periods; females registered a peak at 19:00-21:00hs period and arrived until 03.00; the 75.8% of females and 51.5% males was captured during the 17:00-23:00hs period; and 16.1% of females and 37.08% of males during 1:00-7:00hs period (no significant difference). There was no correlation of T and H with total abundance. When only considered females it was positively correlated with T and negatively with RH in February and March; the higher abundance was between 32.0°-32.9°C and 50.9%-65.9% RH. These results showed a trend in the population seasonality and hourly activity that provides a baseline for the design of protocols on prevention- control strategies research. Keywords: CDC trap, India, Phlebotomus argentipes Increasing distribution of sandfly-borne diseases makes entomological surveillance fundamental to determine changes in vector species and define transmission risks. The CDC light trap (LT) is one of the most commonly used sampling methods for sandflies. On the other hand, CDC gravid traps (GT), normally used for mosquito sampling, have recently been found to collect phlebotomine sandflies in Tanzania and India. In this work we set out to compare the efficacy of the LT vs GT for the collection of sandflies in Bhubaneswar, Orissa. Sampling was carried out using a Latin Square design in the Salia Sahi slum in Bhubaneswar, placing five traps of each type indoors in 10 households during 30 nights (=300 trap-nights). In addition, we tested the efficacy of both traps in a buffalo farm for 24 nights. Sandflies were identified to species, making note of male genitalia rotation and female physiological status (unfed, fed, and gravid). Negative binomial regression was used to analyse the over dispersed catch data and incidence rate ratios were calculated to measure treatment effects. We report the first evaluation of GT for the collection of sandflies, and the first evaluation of this trap indoors. In households, LT was five times more likely to collect sandflies than the GT, nearly five times more likely to collect females (including gravid females), and three times more likely to collect males. Both traps performed similarly in the buffalo farm. The sandfly species identified include Phlebotomus argentipes, Sergentomyia antennata, S. babu and S.squamipleuris. Our results show the LT outperformed the GT in all aspects, and remains one of the best options for entomologicalsurveillance of sandfly populations. We report the intradomiciliary presence of Phlebotomus argentipes in Orissa state. This sandfly vectors Leishmania donovani, which causes anthroponotic visceral leishmaniasis in India. Although no VL cases are reported in Orissa, frequent human migration is a potential threat to the introduction of the parasite into the state from the well-known endemic areas of Bihar, Jharkhand or West Bengal. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 83 ID 47-P Taxonomy & Phylogeny ID 48-P Taxonomy & Phylogeny TAXONOMY AND DIAGNOSTIC CHARACTERS OF THE PHLEBOTOMUS ARGENTIPES SPECIES COMPLEX DISCRIMINANT ANALYSIS AS A POSSIBLE TOOL TO I D E N T I F I C AT I O N S A N D F L I E S ( D I P T E R A : PSYCHODIDAE) SPECIES THROUGH MORPHOMETRIC DATA Kandan Ilango Zoological Survey of India Zoological Survey of India Ministry of Environment & Forests Government of India Southern Regional Centre Chennai, INDIA [email protected] Keywords: Phlebotomus argentipes, taxonomy, species complex, visceral leishmaniasis, India Visceral Leishmanisis (VL) (Kala-Azar) transmitted by the bite of sand fly, Phlebotomus argentipes sensu lato is endemic in parts of India, Nepal and Bangladesh accounting for 90% of the global disease burden. Ph. argentipes s. l. occurs widely in southern Asia exceeding the limits of disease endemic regions but its population's exhibit differences in biology and behaviour. The taxonomy of Ph.argentipes s.l. based on descriptive morphology and principal component analysis reveals unequivocally 3 sibling species viz. Ph. argentipes sensu stricto, Ph. annandalei and Ph. glaucus from India. It remains however unknown which of the three sibling species is the Indian VL vector. A morphologically distinct population of Ph. argentipes from Sri Lanka that differs from the members of Indian species complex of Ph. argentipes exists in areas of active cutaneous leishmanisis transmission. Molecular identification of the Sri Lankan Ph. argentipes using sequences of COI, cyt b, ITS2 and 28s rDNA confirmed the existence two sibling species. Vector control in Indian VL elimination program needs urgently the identification and characterization of members of the Ph. argentipes species complex. Rodrigo E. Godoy1; Eunice A. B. Galati2, Nataly A. de Souza1, Thiago V. dos Santos3; Lindemberg Caranha de Sousa4; Adriana Zwetsch1; Elizabeth F. Rangel1 1 Instituto Oswaldo Cruz - Fundação Oswaldo Cruz; 2Faculdade de Saúde Pública – Universidade de São Paulo; 3 Instituto Evandro Chagas; 4Secretaria de Saúde do Estado do Ceará [email protected] Keywords: Sand flies, Taxonomy, linear morphometrics The sand flies taxonomy based in the linear morphometry and morphology of the adult insect still presents some problems as for the determination of species, especially regarding the females specimens, where more than 150 from a total of 500 species needs the presence of a male specimen to fully determinate the right species. Besides that the presence of species complex it is a further complicating factor for the identification in some genera. Thus our work aimed to use discriminant analysis through morphometric data to evaluate the capacity of this analysis to properly determinate the sand flies species. We evaluate five sand flies species (40 specimens for each species), that are proven Leishmaniasis vectors, where: Nyssomyia intermedia, Ny. whitmani, Bichromomyia flaviscutellata, Migonemyia migonei, Ny. neivai, Brumptomyia troglodytes (used as control group). To cluster the species through the morphometric data we performed 2 different analyzes, one with the characters present both in male and female (shared characters) more the specifics from male (120 specimens) using 48 characters, and the other with the shared characters more the specifics from female (120 specimens) using 43 characters. Two statistical tests, canonical variables and discriminant function were used to obtain the best clustering among species groups. In this analysis, also the values of F and Wilks' Lambda, were evaluated. The characteristics that most contributed to the species differentiation in the 110 males specimens were the structures of the terminalia. All the species presented 100% of correct classification even for the cross-validation, nevertheless between Ny. intermedia and Ny. neivai, after the cross-validation, the correct classification was respectively: 85% (17/20 specimens) and 90% (18/20). The characters that most contributed to the differentiation of the species in the 114 females specimens were the characters shared between males and females. All the species presented 100% of correct classification even for the cross-validation. Nevertheless between Ny. intermedia and Ny. neivai, after the cross-validation, the correct classification was respectively: 85% (17/20 specimens) and 62.5% (10/16). The discriminant analysis using morphometric data can be used as an additional tool for the definition of cryptic species, helping to define species. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 84 ID 49-P Taxonomy & Phylogeny ID 50-P Taxonomy & Phylogeny FIRST REPORT OF ABNORMAL NUMBERS OF SPERMATHECAE IN A FEMALE PHLEBOTOMUS (LARROUSSIUS) LONGICUSPIS (DIPTERA: PSYCHODIDAE) IN TUNISIA FIRST RECORD OF PSATHYROMYIA P R A D O B A R R I E N TO S I ( L E P O N T, M AT I A S , MARTINEZ & DUJARDIN, 2004) (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) IN BRAZIL Jomaa Chemkhi1, Souheila Guerbouj1, Ikram Guizani1, Afif Ben Salah2 Douglas de Almeida Rocha, Jônatas Barbosa Cavalcante Ferreira, Rodrigo Gurgel-Gonçalves & Andrey José de Andrade 1 Laboratoire d'épidémiologie Moléculaire et Pathologie Expérimentale Appliquée aux Maladies infectieuses, Institut Pasteur de Tunis, Université Tunis El Manar, Tunisia; 2 Laboratoire d'épidémiologie Médicale, Institut Pasteur de Tunis, Université Tunis El Manar, Tunisia. Laboratório de Parasitologia Médica e Biologia de Vetores, Área de Patologia, Faculdade de Medicina, Universidade de Brasília, Campus Universitário Darcy Ribeiro, Asa Norte, 70910-900, Distrito Federal, Brazil. [email protected] [email protected] Keywords: Phlebotomine sand fly, spermathecae, abnormality Keywords: Psathyromyia runoides, P. prabobarrientosi, geographical distribution, taxonomy Phlebotomine sand flies (Diptera: Psychodidae) constitute a large group of flies including about 800 species, many of which have been described as vectors of leishmaniasis, a disease caused by Leishmania (Kinetoplastida: Trypanosomatidae) parasites. Species morphological identification is based for males, on the genitalia properties, while it is based mainly on specific characteristics of the spermathecae and ducts, for female specimens. In Tunisia, a total of 17 species belonging to the genus Phlebotomus and Sergentomyia were described. In this work we report an abnormality found in the spermathecae of a female specimen of the species Phlebotomus (Larroussius) longicuspis. Sand flies were collected in the locality of Sidi Saad, in the governorate of Kairouan, central Tunisia, during the period from June to November 2011. CDC light traps were placed overnight, in different biotopes (inside habitations, outdoors and within animal shelters). Specimens were collected and males were washed, mounted and directly identified under a microscope. Females were either dissected and identified or stored in liquid nitrogen for later identification. Dissection of female sand flies was carried out in sterile saline solution. The last two abdominal segments containing spermathecae and ducts were mounted for morphological identification based on the keys of Lewis (1982) and Léger et al. (1983). During the survey of sand fly fauna in the above mentioned region, 163 flies were morphologically described and the species identified, in the locality of Sidi Saad, within the animal shelters biotope. Among the studied sand fly population we found an abnormal spermathecae in the case of a female specimen identified as belonging to the species Phlebotomus (Larroussius) longicuspis. This specimen presented with three spermathecae. Indeed, an additional spermathecae located in a short bifurcation on the upper extremity of one duct was found. This abnormality is considered rare according to the described anatomy of phlebotomine sand flies. The abnormality found in the female spermathecae of the species Phlebotomus (Larroussius) longicuspis is for the first time described in Tunisia. Morphological abnormalities are important to report in order to avoid erroneous sand fly identification or description of new species. This study received financial support from the Tropical Medicine Research Centers (P50) program (Contract No. RFA-AI-06-006), USA. Psathyromyia pradobarrientosi (Le Pont, Matias, Martinez & Dujardin, 2004) was described based on males from Bolivia and nowadays this is the single record of the species. It is morphologically similar to P. runoides (Fairchild & Hertig, 1953), which was described in Panama. Males of these phlebotomine sand fly species can be distinguished based on the ejaculatory duct length in relation to the pump and its apex. The female of P. pradobarrientosi is undescribed. Recently, two males of sand flies identified as Psathyromyia sp. were collected in forested areas of Brasília, Federal District, Brazil. To access the taxonomic identity of these specimens they were compared with the male of P. runoides paratype deposited in the Coleção do Museu de Zoologia da Universidade de São Paulo and with the original descriptions of P. pradobarrientosi and P. runoides. The collected specimens and the types of P. runoides were measured, drawn and photographed. Males of Psathyromyia sp. presented the following characteristics: 1) the ejaculatory duct was 3 times longer than the ejaculatory apodeme (piston + sperm sac); 2) the apex of the ejaculatory duct was inflated at end; 3) the ventral margin of the basiphallus was 2 times smaller than that of P. runoides; 4) the bristles on the dorsal margin of the paramere were distributed at least on the apical half. Based on these characteristics the males of Psathyromyia sp. were identified as P. pradobarrientosi and this species is registered for the first time in Brazil. Due to morphological similarity between P. pradobarrientosi and P. runoides, the geographical distributions of both species should be revised. We hypothesize that Psathyromyia runoides presents a trans-Andean distribution while P. pradobarrientosi is a cis-Andean species in South America. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 51-P Taxonomy & Phylogeny ID 52-P Taxonomy & Phylogeny RECORDS OF FEMALES PSATHYROMYIA CUZQUENA MARTINS, LLANOS & SILVA, 1975 (DIPTERA, PSYCHODIDAE, PHLEBOTOMINAE) IN NORTHERN BRAZIL ASSESSMENT OF THE TAXONOMIC STATUS OF PHLEBOTOMUS PIFANOI ORTIZ, 1972 (DIPTERA, PSYCHODIDAE, PHLEBOTOMINAE) Priscila Bassan Sábio, Andrey José de Andrade and Eunice Aparecida Bianchi Galati 85 Priscila Bassan Sábio, Andrey José de Andrade and Eunice Aparecida Bianchi Galati Faculdade de Saúde Pública, Universidade de São Paulo Faculdade de Saúde Pública, Universidade de São Paulo [email protected] [email protected] Keywords: Phlebotominae, Sand fly, Psathyromyia cuzquena, Keywords: Phlebotominae, sand flies, Shannoni Complex, Phlebotomus pifanoi Misidentification Psathyromyia cuzquena (Martins, Llanos & Silva, 1975) was described on the basis of two males collected in Pilcopata, Cuzco Department, Peru. It belongs to Shannoni Series and genus and subgenus Psathyromyia. This species was considered by the authors morphologically close to Pa. shannoni (Dyar, 1929) and Pa. souzacastroi (Damasceno & Causey, 1944) and is also distributed in Amazonian Brazil, Colombia and Venezuela. In the identification keys of American sand fly species it has only been possible to identify the male of the Pa. cuzquena, since the female has never been described. However, some fauna surveys have reported the female of Pa. cuzquena in northern Brazil, along with other species of the Shannoni Series. These studies were conducted in the municipalities of Borba, Maués, Nhamundá and Manaus, Amazonas state, and Caroebe, Roraima state. Literature searches were undertaken in the Medline, Scopus and SciELO online databases for records of the description of the female of Pa. cuzquena, and some depository collections of species sand fly were consulted, but nothing was found. Sand fly are a group of insects of great importance to public health due to the haematophagic habit of their females that bite human beings and other animals and can transmit infective agents, such as trypanosomes, especially the various species of the genus Leishmania. Misidentifications of species may lead to false geographical records, thus confounding the understanding of the role of the vectors involved in the transmission of diseases. Psathyromyia shannoni (Dyar, 1929) had Phlebotomus pifanoi Ortiz, 1972, known only by its male holotype, considered as its junior synonym – though without any justification – by Martins, Williams and Falcão (1978). Psathyromyia cuzquena (Martins, Llanos & Silva, 1975) was described from a single male, whose female is unknown. The taxonomic status of Ph. pifanoi is here reviewed on the basis of its morphological characters reported in the literature and compared with those of Pa. shannoni and close species deposited in entomological collections and the literature. Psathyromyia shannoni and Ph. pifanoi are morphologically distinct species and therefore not synonymous. In the gonostyle of Ph. pifanoi, the upper external spine is implanted closer to the apical spine than to the external lower spine, while in Pa. shannoni the upper external spine is implanted equidistant from the apical spine and the lower external spine. In the paramere of Ph. pifanoi the bristles on the dorsal edge extend from its apex to the level of the most apical bristle of the ventral margin and in Pa. shannoni the bristles on the dorsal edge extend from its apex to the level of the most basal bristles of the ventral margin. On the other hand, Pa. cuzquena is morphologically identical to Ph. pifanoi. Thus, Psathyromyia pifanoi stat. rev.; comb. nov. is a valid species and in accordance with the criterion of priority it is the senior synonym of Pa. cuzquena sin. nov. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 86 ID 53-P Taxonomy & Phylogeny ID 54-P Taxonomy & Phylogeny UNCHARTED PHLEBOTOMINE SAND FLIES IN GABON MORPHOMETRIC STUDY ON POPULATIONS OF PSYCHODOPYGUS COMPLEXA AND PS. WELLCOMEI ( D I P T E R A : P S Y C H O D I D A E ) , V E C TO R S O F CUTANEOUS LEISHMANIASIS IN BRAZIL Nil Rahola IRD MIVEGEC unit (France) & CIRMF UEM unit (Gabon). [email protected] Keywords: new species taxonomy update Patrick Feitosa Botelho¹, Fábio Márcio Medeiros da Silva¹, Rodrigo Espíndola Godoy², Thiago Vasconcelos dos Santos¹ 1 Instituto Evandro Chagas Svs Ms, Ananindeua, Pa, Brazil [email protected] Despite the former mention of a native case of visceral leishmaniasis, Gabon does not currently constitute an endemic country for this disease. As a result, the sandfly fauna in this country remains poorly documented and prospected. An exhaustive review of literature reports only four species in Gabon. From 2012 to the end of 2013 we managed to collect more than 5000 sand flies through a large mosquito survey that consisted in 1850 CDC miniature light traps installed in two different forest sites and 560 CDC miniature light traps installed in two different caves. Over 1500 specimens were found in forest sites and 3500 in caves. In addition, some collections were performed using other CDC miniature light traps in anthropic environments such as small savannahs villages, forest villages and towns. Those collections allowed us to bring the current checklist for Phlebotomine sand flies of Gabon from four to 25 species, and add at least three new species for Science (currently being described). The forest was the highest spot for species diversity and allowed us to discover and describe a new species of Phlebotomus with a quite singular morphology: Phlebotomus (Legeromyia) multihamatus and also the unknown male of Spelaeomyia moucheti. This discovery will lead us to perform a molecular analysis of the whole genus Spelaeomyia. On the other hand, in caves, sand flies were very abundant but of poor diversity. With those new collections we will be able to bring new morphological data and make some re-descriptions of some specimens such as the female of Spelaeomyia moucheti or Sergentomyia lumsdeni, but also to consider the possibility of the creation of new subgenus for the genus Sergentomyia. This update of the Phlebotomine sand flies of Gabon must encourage prospecting in this country. Even if only one case of leishmaniasis has been reported in Gabon, the role of sand flies in the transmission of such parasites or other infectious agents (such as viruses or bacteria), to wildlife, domestic animals or to humans, as well as their trophic preferences, should be evaluated. Keywords: Morphometrics, Psychodopygus, cutaneous leishmaniasis The correct identification of sand fly species is an important component for strategies of surveillance and leishmaniasis control. Despite medical relevance, the females of Psychodopygus from chagasi series are so far morphologically indistinguishable. Among these species, Psychodopygus wellcomei and Ps. complexa are respectively incriminated and suspected in the transmission of cutaneous leishmaniasis at Amazonian Brazil. The aim of this work is to analyze different populations of Lu. (Ps.) complexa and Lu. (Ps.) wellcomei by classical and geometric morphometry and thus attempt to discriminate them. It was selected 104 individuals from the “Coleção de Flebotomíneos do IEC” (ColFleb IEC) mounted on glass slides with berlese liquid and distributed as follow: Ps. complexa from Vitória do Xingu- Pará State (PA) (20 Males+ 20 Females), Ps. wellcomei from Parauapebas- PA (25M+ 25F), Ps. wellcomei from Baturité- Ceará State (CE) (14M+ 14F). Their correct identification was ensured for the absence of simpatry on the collects. An external group was composed by individuals of Brumptomyia sp. from Ananindeua- PA (3F). The measurements were made with a computer- assisted system (Axiovision- Zeiss). For classical morphometry, 15 characters were selected for females and 19 for males. For geometrical morphometry, 5 anatomic marks were used from the right wing of each specimen. The analysis was made with canonical variables and discriminant function. The classical morphometry allowed distinguish the species with 89,8% of correct classification for males and 69,4% for females at cross validation (88% for Ps. complexa PA, 40% for Ps. wellcomei PA and 66.7% for Ps. wellcomei CE). The geometric morphometry showed short distance (3,0044) between the Psychodopygus spp. in comparison of the external group (10,0677). The female specimens were distinguished with 93, 3% of correct classification in cross validation. In conclusion, although a high probability of correct identification was achieved with geometric morphometry, not all females can be distinguished unambiguously with both methods. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 87 ID 55-P Taxonomy & Phylogeny ID 56-P Taxonomy & Phylogeny TAXONOMIC REVIEW OF PSYCHODOPYGUS MANGABEIRA, 1941 (DIPTERA, PSYCHODIDAE, PHLEBOTOMINAE) TAXONOMIC REVISION OF THE SUBGENUS PSATHYROMYIA BARRETTO, 1962 (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) Veracilda Ribeiro Alves1, Claudio Ruy V. da Fonseca2, Paloma Helena F. Shimabukuro1 Priscila Bassan Sábio, Andrey José de Andrade e Eunice Aparecida Bianchi Galati 1 Centro de Referência Nacional e Internacional de Flebotomíneos/Centro de Pesquisas René Rachou/FIOCRUZ, Belo Horizonte-MG, Brazil. 2 Coordenação de Biodiversidade/Instituto Nacional de Pesquisas da Amazônia/INPA, Manaus-AM, Brazil. Faculdade de Saúde Pública, Universidade de São Paulo [email protected] [email protected] Keywords: Phlebotominae, Psathyromyia, Sandfly, Taxonomy Keywords: Morphological characters, Psychodopygus, sand flies; Neotropical region Phlebotominae sandfly (Diptera, Psychodidae, Phlebotominae) are a group of public health concern due to their haematophagic habit of females that bite man and other animals, causing them annoying and they can also transmit infection agents such as trypanosomes, especially for the various species of the genus Leishmania, bacteria and viruses. Identification of American sandfly is traditionally based on morphology. However some species are morphologically very similar, leading to errors in identification, as in the species included in the subgenus Psathyromyia. Because of this similarity, especially among females, this subgenus had some of their species considered as junior synonyms of Psathyromyia shannoni. Some of these were revalidated in a previous study. Thus, this project aims to review the taxonomic status of the species of the subgenus Psathyromyia by means of morphological, morphometric and molecular analyzes. The use of molecular studies, together with morphological can help in the correct diagnosis of species of this subgenus. With this, we hope to contribute to a better understanding of the role epidemiological and vector of these species in Brazil. The species of the genus Psychodopygus Mangabeira, 1941 occur in woods and forests. In human bait collections carried out in the Amazon region at the forest floor level, most species of sand flies collected belong to this genus. The females of some species are difficult to identify due to the existing interspecific morphological similarity, therefore correct identification is only possible when they are collected together or in copulation. The species are predominantly neotropical occurring from southern Mexico to southern Brazil with over 80% of the species recorded in the Amazon region. The species of this genus are of medical interest because they are vectors of Leishmania and are highly anthropophilic. The revision of the species was based on the examination of type specimens deposited in the following biological collections: Faculdade de Saúde Pública(FSP)/SP, Instituto Evandro Chagas (IEC)/PA, Museu de Zoologia da USP (MUSP)/SP, Instituto Oswaldo Cruz (IOC)/RJ, Instituto Nacional de Pesquisas da Amazônia (INPA)/AM, Centro de Pesquisas René Rachou CPqRR)/MG, Walter Reed Biosystematics Unit (WRBU)/Smithsonian Institute/USA and Institut de Recherché pour le Développement (IRD)/France. At the beginning of this study the genus Psychodopygus comprised 39 valid species and of these 24 type-species were examined. The types of nine species could not be examined due to difficulties in contacting those responsible for the collections in which they were deposited; six types have been lost: Ps. fairchildi (Barretto, 1966), Ps. guyanensis (Floch & Abonnenc, 1941)¸ Ps. hirsutus (Mangabeira, 1942), Ps. complexus (Mangabeira, 1941), Ps. geniculatus (Mangabeira, 1941) and Ps. parimaensis (Ortiz e Álvares, 1972). The informally described species of Três Esquinas morphotypes were confirmed as being Ps. guyanensis. Psychodopygus douradoi (Freitas & Fé, 1998) is proposed as junior synonynym of Ps. fairtigi (Martins, 1970), and Ps. parimaensis as a junior synonynym of Ps. davisi (Root, 1934). Thus, the number of valid species after this study is 36. This revision revealed the status of the type species deposited in different institutions and the importance of examining them for comparison with other similar species and descriptions of new species. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 57-P Taxonomy & Phylogeny ID 58-P Taxonomy & Phylogeny DNA BARCODING FOR THE IDENTIFICATION OF SAND FLY SPECIES (DIPTERA, PSYCHODIDAE, PHLEBOTOMINAE) IN PORTUGAL NEW RECORDS OF PHLEBOTOMINAE (DIPTERA: PSYCHODIDAE) FROM THE ATLANTIC FOREST REGION OF ARGENTINA C. Maia1,2,3, R. Parreira4,5, J. Cristóvão 1,2, M.O. Afonso1,5, L. Campino1,6 Casertano,S.A.1; Andrade Filho, J.D2; Salomon, O.D1,3 1- Unidade de Parasitologia Médica, Instituto de Higiene e Medicina Tropical (IHMT), Universidade Nova de Lisboa (UNL), Lisboa, Portugal 2-Centro de Malária e outras Doenças Tropicais 3-Faculdade de Medicina Veterinária, Universidade Lusófona de Humanidades e Tecnologias, Lisboa, Portugal 4-Unidade de Microbiologia Médica, IHMT, UNL 5-Unidade de Parasitologia e Microbiologia Médicas 6-Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, Faro, Portugal [email protected] Keywords: Phlebotomine sand flies, Barcoding, Cytochrome oxidase gene subunit I, Portugal Phlebotomine sand flies (Diptera, Psychodidae) transmit pathogens that affect humans and animals worldwide. As sand fly genera and species vary in geographical distribution, ecology, and pathogen transmission, surveillance of their presence and distribution are important. Five sand fly species are known to be present in Portugal: Phlebotomus ariasi, P. papatasi, P. pernicious, P. sergenti and Sergentomyia minuta. P. perniciosus and P. ariasi are the proven vectors of Leishmania infantum while L. major DNA was recently detected in one S. minuta specimen collected in the south of the country. Although the identification of sand flies is currently based on morphological characters, it requires considerable skill and taxonomic expertise. DNA-based approaches are useful tools for estimating sand fly diversity and for ensuring an accurate identification of species. A partial sequence of the mitochondrial cytochrome oxidase gene subunit I is currently being used to differentiate species in different animal taxa, including insects. The aim of this study was to evaluate the utility of the DNA barcoding for the identification of phlebotomine sand flies in Portugal. P. ariasi, P. pernicious, P. sergenti and S. minuta specimens collected from the North, Center and South of the country were morphologically identified prior to their use for barcoding. Analyses of the sequences generated indicated that the observed species clusters were consistent with the morphological identifications. In conclusion, barcoding is a useful tool for discriminating the sand fly species existent not only in Portugal but also in the Mediterranean basin, and in the future can probably be used as an extra and efficacious tool for monitoring the presence and spread of the different sand fly species responsible for the transmission of pathogens of medical and veterinary concern. Funding: This work was partially supported by EDENext (FP7INCO-CT-2010-261504). C. Maia (SFRH/BPD/44082/2008) holds a fellowship from Fundação para a Ciência e a Tecnologia, Ministério da Ciência, Tecnologia e Ensino Superior, Portugal. 88 1 Instituto Nacional de Medicina Tropical - INMeT, Puerto Iguazú Argentina,REDILA; Centro de Pesquisas René Rachou - FIOCRUZ, Belo Horizonte, Minas Gerais, Brazil; 3 CONICET, Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina 2 [email protected] Keywords: new records, Argentina, phlebotomine species, Atlantic Forest The Atlantic Forest is one of the most endangered rainforests on earth, of which remains about 7% of its original extension. Misiones Province in Argentina and neighboring areas of Brazil and Paraguay contain the “Green Corridor”, the largest remnant of this forest that, together with the populated areas, also constitute one of the most important geopolitical, economical and cultural cross points of South America. Recent increasing sampling efforts in studying Phlebotomine sand flies communities in different anthropic and nonanthropic environments of the Atlantic Forest of Argentina have resulted in new records for the region. Four species have been found in natural protected areas (Iguazú National Park) and in forest patches remaining in recently occupied areas, both in Puerto Iguazú municipality, north of Misiones Province: Micropygomyia capixaba (Dias, Falcão, Silva & Martins, 1987); Evandromyia correalimai (Martins, Coutinho & Luz, 1970); Evandromyia edwardsi (Mangabeira, 1941) and Evandromyia bourrouli (Barreto & Coutinho, 1941). In rural environments nearby San Pedro city, central Misiones Province mountain area, Martinsmyia alphabetica (Fonseca, 1936) was found, more than 60 years after its last record. The main landscape out protected areas consisted in little farms containing small patches of rainforest. All these species have been also found in neighboring Brazilian State of Paraná in the same semidecidual rainforest, and M. alphabetica have been also reported in urban areas of Brazilian neighboring city of Foz do Iguaçú. These results, together with recent findings in Chaco Region, increase Argentina's Phlebotomine species number up to 36. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 89 ID 59-P Taxonomy & Phylogeny ID 60-P Taxonomy & Phylogeny A METHOD FOR EXTRACTION OF GENETIC MATERIAL WHICH PERMITS THE INTEGRATION OF CLASSICAL TAXONOMY WITH IDENTIFICATION BASED ON THE DNA BARCODE CONTRIBUTION TO THE KNOWLEDGE OF THE FAUNA OF SAND FLIES (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) OF RIO DOCE STATE PARK, MINAS GERAIS, BRAZIL Alveiro Pérez-Doria, Luís Romero-Ricardo, Natalia Lastre, Eduar Elías Bejarano Cristian Ferreira Souza1; Reginaldo Peçanha Brazil2; José Dilermando Andrade Filho3; Paula Dias Bevilacqua4; Patrícia Flávia Quaresma3; Mirelle Barbosa Souza5 Grupo de Investigaciones Biomédicas, Universidad de Sucre, Sincelejo, Colombia. [email protected] Keywords: DNA Extraction, Sandflies, classical taxonomy, DNA barcoding In the study of phlebotomine sand flies molecular biology techniques have provided valuable evidence that has helped resolve taxonomic problems both at the infraspecific and supraspecific levels. Nevertheless, in most of these studies part or even all of the insect body is used to extract the genetic material which is subsequently used for genotyping. Thus, although analysis of the sequences generated may resolve the research question, the voucher specimens required for classical taxonomy remain incomplete due to the loss of morphological characters. With this problem in mind, in the present study the concentration and quality of the DNA obtained with a simple extraction method based on high salt concentrations and enzymatic maceration were evaluated, this technique yielding the genetic material of the insect while allowing the exoskeleton to be recovered intact for use as a voucher specimen. Phlebotomines were collected in Colosó, in the Colombian departament of Sucre. The effects of different concentrations of proteinase K and distinct incubation times on the final concentration of DNA were determined by measuring in a spectrophotometer. Integrity of was evaluated by amplifying and sequencing a 550 bp fragment of mitochondrial DNA. The DNA concentrations varied between 12 and 236 ng/ l, levels that were markedly higher than those obtained for the group processed by mechanical maceration (15 - 42 ng/ l). The barcode sequences of the cytochrome b gene of Lu. evansi, Lu. panamensis, Lu. gomezi, Lu longipalpis, Lu. dubitans, Lu. trinidadensis, Lu. micropyga, Lu. cayennensis cayennensis, Lu. venezuelensis and Lu. atroclavata were obtained. The exoskeletons were recovered intact and mounted in Canada balsam. These results provide entomologists with a useful tool for the morphological and genetic identification of species, permitting the integration of classical and molecular taxonomy. 1 Programa de Pós-Graduação em Biologia Parasitária - Instituto Oswaldo Cruz Fundação Oswaldo Cruz – Av. Brasil, 4365 – Manguinhos, CEP: 21040-360. Rio de Janeiro–RJ. Brazil; 2Laboratório de Doenças Parasitarias - Instituto Oswaldo Cruz Fundação Oswaldo Cruz - Manguinhos,RJ. Brazil; 3Centro de Referência Nacional e Internacional para Flebotomíneos - Centro de Pesquisas RenéRachou/Fiocruz, Av. Augusto de Lima, 1715 - Barro Preto, CP1743. CEP: 30190-002. Belo Horizonte–MG. Brazil; 4Departamento de Veterinária - Universidade Federal de Viçosa, Av. Peter Henry Rolfs, s/n - Campus Universitário. CEP: 36570-000. Viçosa–MG. Brazil; 5 Graduação em Ciências Biológicas - Universidade Federal de Viçosa, Av. Peter Henry Rolfs, s/n - Campus Universitário. CEP: 36570-000. Viçosa–MG. Brazil. [email protected] Keywords: sand flies, vectors, Leishmaniasis Currently about 500 species of sand flies are known in the Americas, and only in Brazil have been recorded 267 of these species. This study aimed to identify the sand fly fauna found at the Parque Estadual do Rio Doce (PERD), located in the eastern state of Minas Gerais, Brazil, between the municipalities of Timóteo Marliéria and Dionísio. The collections were made in Juquita Trail, located in the PERD area belonging to the municipality of Timóteo-MG, and the sand flies of the urban area of the municipality is already known. During collection 12 CDC light traps, installed from September 2012 to February 2014 were used. Traps were exposed for two consecutive nights, at a height of 150 cm from the floor nights. To date a total of 994 sand flies belonging to 20 species were identified, 693 (69.7%) male and 301 (30.3%) females, were: Brumptomyia avellari, Brumptomyia cunhai, Brumptomyia nitzulescui, Evandromyia cortelezzii, Evandromyia sallesi, Evandromyia termitophila, Evandromyia tupynambai, Micropygomyia borgmeirai, capixaba Micropygomyia, Micropygomyia quinquefer, Migonemyia migonei, Nyssomyia intermedia, Nyssomyia Whitmani, Pintomyia fischeri, Pintomyia pessoai, Pressatia choti, Psathyromyia lanei, Psathyromyia pascalei, Psychodopygus carreirai, Psychodopygus davisi, Sciopemyia sordellii and Trichopygomyia longispina. Among the species caught Pressatia choti (21.3%) Trichopygomya longispina (16.1%) and Nyssomyia intermedia (14.2%) were the most abundant species. It is very important to report the presence of Nyssomyia whitmani (5.2%), Migonemyia migonei (3.0%), and Nyssomyia intermedia species already recognized as vector of Leishmania. Importantly, the PERD is located in an endemic area of leishmaniasis, with cases already recorded among employees and park visitors. The identification of these species as possible vectors, help to better understand the cycle of transmission of leishmaniasis within the Park boundaries. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 90 ID 61-P Taxonomy & Phylogeny ID 62-P Taxonomy & Phylogeny SAND FLY FAUNA IN KALEYBAR DISTRICTS, NORTHWESTERN IRAN MALFORMATIONS OF THE GENITALIA IN MALE MICROPYGOMYIA QUINQUEFER (DYAR, 1929) (DIPTERA: PSYCHODIDAE) IN CORRIENTES, ARGENTINA Amini Esfanjani MH1, Barazesh A1, Mazloumi Gavagni A1,SaeidiZ 2* 1 Department of Medical Parasitology, Faculty of Medicine, Research Center for Infectious and Tropical Medicine, Tabriz University of Medical Sciences, Tabriz, Iran. 2 Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. [email protected] Mirta Liliana Mierez, María Josefa Felisa Rea, Carlos Borda, Luis Armando Mosqueda Edgardo Centro Nacional de Parasitología y Enfermedades Tropicales (Cenpetrop), Facultad de Medicina, Universidad Nacional del Nordeste, Santa Fé 1432, 3400 Corrientes, Argentina [email protected] Keywords: visceral leishmaniasis, sand fly, Kaleybar, Iran Introduction: In Iran Leishmania infantum is the main causative agent of the visceral leishmaniasis (VL). Studying on vector species is valuable for epidemiological studies and for planning control mesures. So this study was carried out to diagnose and determine the sand fly species in eight villages of Kaleybar. Methods: Four VL endemic villages and four none endemic villages were selected from Kaleybar district, light traps and in some occasions sticky papers were used to collect sand flies. They were mounted using lactophenol and Pouri's media and identified by the use of valid identification keys. Results: A total of 5000 sand flies were collected and 500 sand flies were identified. The study showed that subgenus Larroussius of Phlebotomus, in particular P. perfiliewi, P. tobbi and P. kandelakii, among endemic villages and P. (Paraphlebotomus) sergenti among none endemic villages were the most abundant. Discussion: It seems that P. (Larroussius) perfiliewi was the most abundant species in the VL endemic areas and P. (Larroussius) tobbi and P. (Larroussius) kandelakii were in second rank. Other studies in Northwestern endemic foci of Iran also reported similar results. Such researches would be crucial in this area because there are some nomadic tribes which maybe could play a role to transfer the disease to different areas with various climates. Keywords: Malformations, male, Micropygomyia quinquefer, Argentina A preliminary study indicated a high prevalence of dermal leishmaniasis in the human population of the Corrientes province. Based on the reported cases of human infections of cutaneous leishmaniasis, the rural district of Itatí was selected for the entomological studies. Nyssomyia neivai, Migonemyia migonei and Micropygomyia quinquefer were collected in a sylvatic habitat of Itatí (Borda et al. 2006).The predominance of Mi. quinquefer and its abundance in this area agree with other studies that indicated that this species is in the process of adapting to modified environments especially during the day. Mi. quinquefer belong to a group that was described with five well-developed spines on the style, of which two are terminal, among other morphological characteristics. Morphological characters present in the genitalia of phlebotomine are essential for correct identification of species. The taxonomic identification of the Phlebotominae associated with the epidemiological cycle of leishmaniasis is based on morphological characters of the adult insect. Changes in anatomic structures of taxonomic importance may eventually induce to errors in the species identification. The aim of this study is to report the finding of anomalies in male terminalia of Mi. quinquefer captured in a primary forest in Itatí department, situated on the left coast of Parana River to 75 km from Corrientes city where patients of (ATL) inhabited. Sand flies were collected, with automatic light traps (CDC type) and using manual captures. The collections were conducted during four months of 2010 (April, September, November and December) and 2011 (January, February, Mars and April). The classification system used was that of Young y Duncan (1994) and Galati (2003). A total of 3,563 Mi. quinquefer was captured (1,520 males and 2,043 females). In 2010 643 males were captured and asymmetrical anomalies in the number of spines of style were observed in 0.9% of the collected males. Herein, we described the morphological anomalies Two males had an supplementary spine implanted in the middle in a style Four males had a supplementary terminal spine on one style. As these structures are paired, examination of the corresponding structure on the opposite side of the body normally permits correct identification. It is important to publish this finding, because new taxa can be described from anomalous specimens, leading to confusion among specialists as to the validity of certain species. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 91 ID 63-P Taxonomy & Phylogeny ID 64-P Taxonomy & Phylogeny THE USE OF DNA BARCODING FOR TAXONOMIC IDENTIFICATION OF THE IMMATURE STAGES OF PHLEBOTOMINE SANDFLIES IN OVEJAS, SUCRE (COLOMBIA) SAND FLIES (DIPTERA: PSYCHODIDAE) FROM PERU: REPORT OF NEW RECORDS Roberto Fernandez, Victor Zorrilla, Liz Espada, Gissella Vásquez & Frederick Stell Laura Posada-López1,2; Rafael Vivero1,2; Sandra I Uribe1,2; Luz A. Acosta1; Andrés Vélez-Mira1; Horacio Cadena1; Luis G Estrada3; Edgar Ortega3; Eduar Bejarano3; Ivan D Vélez1 NAMRU-6 Lima-Peru. [email protected] 1 PECET-Programa de Estudio y Control de Enfermedades Tropicales, Universidad de Antioquia, Medellin-Colombia; 2GSM Grupo de sistemática molecular, Universidad Nacional sede Medellin-Colombia; 3Grupo de Investigaciones Biomédicas, Universidad de Sucre, Sincelejo, Colombia [email protected] Keywords: Barcode, inmature, natural breeding Introduction: Phlebotomine sandflies studies in endemic areas of leishmaniasis, usually only include the collection and identification of adults, ignoring the importance searching and identifying the immature stages to confirm the presence of all potential vector species. This problem arises from the difficulty of finding breeding sites and the lack of taxonomic keys for immatures. The mitochondrial cytochrome oxidase I gene (COI) has been successfully used as a DNA barcode for the molecular identification of insect species of medical importance, making it ideal for the identification of immature phlebotomine sandflies. In this study we aimed to validate the use of the COI gene (5'end) as a tool (barcode) for the identification of immature phlebotomine sandflies isolated from natural breeding sites. Methodology: Adult and immature stages of sandflies were collected in Ovejas, a cutaneous and visceral leishmaniasis foci in the Colombia Caribbean Coast on 2013. Adults were identified by morphology using the taxonomic key of Young & Duncan 1994. We extracted total DNA from phlebotomine sandflies larvae and pupae, amplifying a product of 700 bp from the COI gene, using the adults order to compare the sequences (n=17, eight species). We evaluated patterns of nucleotide variability, calculated the genetic distance K2P (intra and interspecific), and generated a dendrogram using neighbor joining. We used our database of sequences obtained from adult Lutzomyia in Colombia, which consists of 36 species (148 sequences), to verify identity. Result: We obtained sequences of the 700bp fragments of the COI mitochondrial gene, from larvae (n=15) and pupae (n=2). The identity of the immature stages corresponds mainly to Lu. evansi and Lu. atroclavata, which was determined by comparison with the sequences of adults previously identified to species by morphological taxonomy. The percentages of variation of Lu. evansi were 1.7% in larvae and 2.3% in adults. We found groups with brackets of 99% for larvae and adults of Lu. evansi and other species identified with adults (Lu. cayennensis, Lu. atroclavata, Lu. micropyga, Lu. rangeliana, Lu. gomezi, Lu. shannoni and Lu. dubitans). Conclusion: The COI gene is a suitable molecular marker for the identification in adults and immature sand flies. High nucleotide variability in adult and immature in Lu. evansi in the same location was detected, however the rates of divergence are within the proposed range for intraspecific difference. Keywords: Sand flies, Peru Phlebotomine sand flies are distributed in tropical and sub-tropical regions of the world, where some species are involved in the transmission of human pathogens including bacteria, parasites and arboviruses. In Peru, sand flies are distributed in the occidental slopes of the Andes as well as in lowland areas in the Amazon Region. Currently, the New World sand fly fauna comprises 512 species distributed in many taxonomic groups. Since 1986, we have been conducting sand fly surveillance in Peru with the goal of characterizing the sand fly fauna and sand fly species distribution in different ecological environments, and identifying the sand fly vectors of leishmaniasis and bartonellosis. Using internal and external morphological characteristics of phlebotomine sand flies we have identified 188 species distributed in two genera, Lutzomyia and Brumptomyia, and a total of 13 subgenera, 8 species group and 5 nongrouped species. In this report we include 15 new records for Peru, all found in the Amazon Region: L. monstruosa, L. pinottii, L. falcata, L. araracuarensis, L. cultellata, L. spathotrichia, L. ferreirana, L. christenseni, L. duncanae, L. beniensis, L. sp de Tarapoto, L. gantieri, L. guilvardae, B. beapertuyi, and B. leopoldoi. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 92 ID 65-P Taxonomy & Phylogeny ID 66-P Taxonomy & Phylogeny FIRST RECORD OF THE GENUS PINTOMYIA COSTA LIMA, 1932 (DIPTERA PHLEBOTOMINAE) IN THE NORTHWEST OF ARGENTINA MORPHOMETRIC STUDY OF LUTZOMYIA L O N G I PA L P I S P O P U L AT I O N S ( D I P T E R A : PSYCHODIDAE) COLLECTED IN URBAN AND ENVIRONMENTAL PROTECTED AREAS IN MINAS GERAIS, BRAZIL Fuenzalida AD1,2,3, Direni Mancini JM1,2, Salomon OD1,2,4, Quintana MG1,2,3,4 1 Reis, A. S., Saraiva, L., Gontijo, C.M.F., Andrade Filho, J. D. Instituto Nacional de Medicina Tropical – MSN, 2 REDILA, 3 Instituto Superior de Entomología – UNT, 4 CONICET. Centro de Pesquisas René Rachou/FIOCRUZ [email protected] [email protected] Keywords: Pintomyia, Jujuy, Tucumán, Argentina Keywords: Lutzomyia longipalpis, Leishamaniasis, Taxonomy, Phlebotominae Currently 30 species of sand flies from 13 genera have been recorded in Argentina. Whithin the genus Pintomyia, seven species have been recorded in four Eastern provinces of the country: Pintomyia bianchigalatiae (Andrade-Filho, Aguiar, Dias & Falcão, 1999), Pi. damascenoi (Mangabeira, 1941), Pi. fischeri (Pinto, 1926), Pi. misionensis (Castro, 1959), Pi. monticola (Costa Lima, 1932), Pi. pessoai (Coutinho & Barretto, 1940) and Pi. torresi (Le Pont & Desjeux, 1991). As part of research project of which objective is to validate distribution maps of different Phlebotominae species, capture were performed in areas with no previous records, between March and May 2013. In Jujuy (24º 11' 04.13”, 65º 18' 02.54”) six sample sites were select, and eight sites in Tucumán province (26º 48' 28.69”, 65º 13' 09.09”), both corresponding to the transition area of Yungas rain forest and dry Chaco. Captures were made with mini CDC like lighttraps. In Jujuy province a total of 32 individuals were caught, which belong to Migonemyia migonei (França, 1920) (47%), Nyssomyia neivai (Pinto, 1926) (25%), Pintomyia sp. (16%), y Evandromyia cortelezzii (Brèthes, 1923) (12%). In Tucumán 270 individuals were collected Mg. migonei (77%), Pi. sp. (18%), Ev. cortelezzi (5%). The presence of Pintomyia is recorded for the first time in to northwest province of Argentina. Pintomyia torresi were recorded to date, only in the province of Chaco; being aware of the relationship of this species feeding on bats, it is noted that the capture individuals belong to site in the presences of bats, and in the case of Tucumán province, the sample area presents the third greatest colony of insectivorous bats of the South America (Tadarida braziliensis). The identified samples of this work, belonging to genus Pintomyia (44 female) and (11 males) were not identified a specific level, although the studied in morphologic characters allow it to be included in the genus and subgenus Pintomyia (Pifanomyia). The morphologic similarities to Pi. torresi in the case of the female genitalia characters , but they are different in the cibarium characters; in the case of males related to this female the differences are more precise, especially in structures associated to the paramere. Due to all above mentioned, a deeper studies will extended in order to determine whether this is a new species for Science. Leishmaniasis are a serious public health problem in Brazil. Belo Horizonte and its metropolitan region present high incidence and mortality due to Visceral Leishmaniases. Lutzomyia longipalpis is the main vector of Leishmania infantum in the country. In the present study morphological aspects of 30 specimens from an urban area, located in Belo Horizonte (Venda Nova – Sanitary District) and 30 specimens from an environmental protected area, the Sumidouro State Park distant 50 km from Belo Horizonte were analyzed. Samplings were conducted from August 2011 to August 2013, using CDC traps during three consecutive days. The specimens were prepared, assembled and submitted to morphometric measurements. The comparisons between males from the two study populations, demonstrated statistical significant differences in mean measures of six characters (length of Labrum-epipharynx, P1, P4, P5, length gamma and length of lateral lobe) from the 52 analyzed. To the females, significant differences were observed in the mean of nine characters (AIII, AXVI, AIV, P1, P3, P4, length gamma and length of spermathecae) of the 46 analyzed. Comparing the variance of the two groups the results indicate that the population of the Sumidouro State Park is more homogeneous, while the population of Venda Nova is more heterogeneous taking into account the morphometric measurements. Lu. longipalpis demonstrates ability of adapting to different habitats and the highest morphological variability of the specimens of the species in the urban area may be related to this behavioral plasticity. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 67-P Ecology & Epidemiology ID 68-P Ecology & Epidemiology SAND FLIES CAPTURES AND IDENTIFICATION OF LEISHMANIA SUBGENUS IN GIEMSA-STAINED SLIDES OF PATIENTS FROM FIVE LOCALITIES OF SALTA, ARGENTINA. PHLEBOTOMINAE BIONOMICS IN THE HUMID CHACO BIO-REGION. CHACO, ARGENTINA Almazán María Cristina1, Gil José F3, Chanampa Mariana M3, Barroso Paola A2, Nasser Julio R1, 4, Copa Griselda N1, Oreste F4, Castillo G4 1 Instituto de Investigación de Enfermedades Tropicales, UNSa-Orán; 2Instituto de Patología Experimental, CONICET-UNSa; 3Instituto de Investigaciones en Energía No Convencional, UNSa; 4Cátedra de Química Biológica, Facultad de Ciencias Naturales UNSa. [email protected] Keywords: Lutzomyia, Leishmania, PCR, RFLP Leishmaniasis is a disease which is endemic in 88 countries and in Argentina remains endemic in 9 provinces. It is caused by several species of flagellates parasites of the genus Leishmania. The wide spectrum of clinical manifestations depends on Leishmania spp and the host inmuno-response. The parasite is transmitted by sand flies of Family Psychodidae, Subfamily Phlebotominae. In Argentina, many sand flies have been described, being the most important and suspected vector Nyssomyia neivai because it showed natural infection with Leishmania parasites. In our country some Leishmania species were isolated Leishmania (Viania) braziliensis, Leishmania (Leishmania) amazonensis, L. (V.) guyanensis, L. (V.) panamensis associated to tegumentary leishmaniasis (TL); and L. (L.) infantum associated to visceral leishmaniasis. The aim of this work was to explore the sand flies presence from five localities of the north of Salta. Besides, patient samples from these areas, were analyzed in order to identify the causal agent. Sand flies were collected during one or six nights with CDC light traps. Traps were placed a meter above the soil and left since 18 pm to 7 am. Species identification of all collected sand flies was made by morphology of spermatheca and cibarium or external genitalia. Leishmania subgenus determination was carried out by PCR-RFLP assay. Seventy-six giemsa-stained samples diagnosed for TL in 2002 at Instituto de Investigación de Enfermedades Tropicales located in Orán, Salta were used. DNA extractions were made by a lysis buffer. The PCR primers L5.8S 5 ' T G ATA C C A C T TAT C G C A C T T 3 ' a n d L I T S R n 5'CTGGATCATTTTCCGATG3' were used. For RFLP, amplicons were digested with HaeIII enzyme. We captured 1352 phebotomines, of which Ny. neivai represent the 76%, Migonemyia migonei 12%, Evandromyia cortelezzii complex 11% and Psathyromyia shanonni 1%. Ny. neivai was present in all localities studied. By PCR-RFLP, Viannia subgenus was the only identified. Regarding geographical cases distribution, 97% of them were from Orán, being this area the site with more cases and highest sand flies amount. Three sandflies species found in this study are considered as potential cutaneous leishmaniasis vectors, particularly of Viannia subgenus parasites. We propose PCR-RFLP in order to enhance the traditional diagnostic just because knowing the leishmaniasis causal agents would improve the treatment assignation and we suggest sandflies surveillance in these localities. 93 Szelag Enrique A1,2, Rosa Juan R.1, Parras Matias A.1,2, Salomón Oscar D.2 REDILA 1Instituto de Medicina Regional (Universidad Nacional del Nordeste). Instituto Nacional de Medicina Tropical. 2 [email protected] Keywords: Tegumentary Leishmaniasis, Nyssomyia neivai, Migonemyia migonei In Argentina, Tegumentary Leishmaniasis (TL) was first recorded in 1916. Actually is considered endemic in 9 provinces from north-east to north-west of the country. Urban Visceral Leishmaniasis (VL) produced the first autochthonous human case in Argentina in the city of Posadas in 2006. Currently, transmission is recorded in Misiones, Corrientes, Santiago del Estero, Formosa and Salta. In order to characterize the distribution in space and time of potential vectors of TL and VL; during the period October 2010 - September 2012 samplings were carried out in three areas of the Chaco province: Resistencia, Margarita Belén and Colonia Benítez. In each area an inhabited dwelling was selected, and CDC light traps were installed monthly during one single night from 7pm to 7am next day in domicile, Peri-domicile and forest. Data of Relative Humidity, High and Low Temperature, were recorded. The Phlebotominae captured were clarified and identified according to Galati (2003). A total of 1499 Phlebotominae was captured in this study. Eight species were recorded: Brumptomyia brumpti, Br. avellari, Evandromyia cortelezzii, Ev. sallesi, Migonemyia migonei, Sciopemyia sordelli, Nyssomyia neivai, Psathyromyia shannoni. Predominant species were Mg. migonei(41.03%) followed by Ny. neivai(38.96%) and complex cortelezzii(9.54%). The greater abundance of Phlebotominae was recorded in Forest(n=1003), followed by Peridomicile(n=423) and domicile(n=73). Mg. migonei predominated in summer with an average temperature of 28.09°C and relative humidity of 61%, while in spring the most frequent species was Ny. neivai with averages of 22.5 ºC and 58% RH. The lower abundance of Phlebotominae was recorded between June and July. The increased frequency of these species in warm and dry months should be considered when proposing control measure. In conclusion, the presence of competent TL vectors as Ny. neivai and Mg. migonei (also a potential vector of VL) in the forest and in lower proportion in peridomestic environments, suggested for the region a risk of transmission of leishmaniases, mainly due to human activities related to the forest, but also by periodical colonization of the peridomestic areas with population of vectors from the forest. Further studies should determine for TL the role of Mg. migonei, the main vector in the Dry Chaco region, as an opportunistic vector in hot-spot transmission, and its interaction with Ny. neivai the main vector in the Humid Chaco region of Argentina. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 69-P Ecology & Epidemiology ID 70-P Ecology & Epidemiology DISPERSION OF LUTZOMYIA LONGIPALPIS IN THE CHACO PROVINCE, ARGENTINA MONTHLY DISTRIBUTION OF LUTZOMYIA LONGIPALPIS IN AN ENDEMIC AREA OF VISCERAL LEISHMANIASIS IN SÃO PAULO STATE, BRAZIL Szelag Enrique A.1,2, Parras Matias A. 1,2, Rosa Juan R. 1, Salomón Oscar D.2 REDILA 1Instituto de Medicina Regional (Universidad Nacional del Nordeste) – Instituto Nacional de Medicina Tropical. 2 [email protected] Keywords: VL - Visceral Leishmaniasis - Migonemayia migonei In Argentina Lutzomyia longipalpis, the main vector of Leishmania infantum, was recorded for the first time in 2004 in Formosa province. In the following years, the vector spread to the south and west of the country being recorded in the provinces of Misiones (2006), Corrientes (2009), Chaco, Entre Ríos (2010) and Salta (2013). Given this fact, the aim of this work was monitoring the spread of Lu. longipalpis in a receptive area of Visceral Leishmaniasis (VL) transmission, in the Chaco province. Captures of Phlebotominae were made in the city of Resistencia (27°27´S 58°59'W), based on a stratified systematic sampling method. From November 2010 to May 2012, four strata were sampled once every season, except in winter. The samples were selected by listing and numbering the neighborhoods of each stratum and randomly, selecting 12 neighborhood per season. In each neighborhood, the house considered the “worst case epidemiological scenario” was selected as sample site. At each sample site, insects were captured by light CDC traps located at 1.5 m, above the animal shelter (kennel, henhouse, etc.) in the courtyard of the house, during one single night (from 17.00 to 9.00 h). Urban localities near Resistencia were also selected as sentinel sites sampled one per season, to monitor any eventual spread from Resistencia. These sentinel sites included the villages of Margarita Belén (27°15'S- 58°58'W), Isla del Cerrito (27º17'S- 58°37'W) and Barrio de los Pescadores (27°27'S- 58°51'W).During the study period, 14 out of 85 traps installed had at least one phlebotomine, obtaining a total of 38 Phlebotominae distributed in three locations: Resistencia, Margarita Belen and Barrio de Pescadores. The species recorded were Lu. longipalpis (13.2%), Migonemyia migonei (15.8%), Nyssomyia neivai (39.5%) and cortelezzii complex (31.6%), represented by Evandromyia sallesi and Ev. cortelezzii. In this monitoring Lu. longipalpis was absent in the city of Resistencia, and its presence was restricted to Barrio de los Pescadores, the closest area to the city of Corrientes (with confirmed presence of the vector, VL human and canine cases). In conclusion, this monitoring of Lu. longipalpis showed that the incipient colonization of the VL vector observed in 2010 with captures of low relative abundance in the city of Resistencia, was not followed by continuous installation of vector populations and expansion of their spatial distribution as in other urban centers of Argentina. 94 Fredy Galvis Ovallos1, Anaiá da Paixão Sevá2, Estela Galucci2, Rodrigo Martins Soares3, Fernando Ferreira3, Eunice Aparecida Bianchi Galati4 1 Postgraduate Program in Public Health, School of Public Health, University of São Paulo- USP, Brazil. 2Postgraduate Program in Experimental Epidemiology of Zoonoses, School of Veterinary Medicine and Animal Science, University of São Paulo-USP, Brazil. 3Department of Preventive Veterinary Medicine and Animal Health, School of Veterinary Medicine and Animal Science, University of São PauloUSP, Brazil. 4Department of Epidemiology, School of Public Health, University of São Paulo – USP, São Paulo, SP, Brazil [email protected] Keywords: Visceral leishmaniasis, São Paulo, Lutzomyia longipalpis Visceral leishmaniasis represents a great challenge to public health in Brazil in view of the great, widespread incidence of both human and canine infection. The area affected by visceral leishmaniasis in São Paulo state has been expanding since 1998. Human and canine cases have been registered in 105 municipalities, the presence of the main vector, Lutzomyia longipalpis, having been registered in the urban area of 148 municipalities. This study sought to describe the Phlebotominae fauna in the urban area of Panorama municipality - SP and the monthly frequency of Lu. longipalpis. The sandfly captures were made from July 2012 to April 2014 with automatic light traps. Two traps were installed in the intra and peridomicile of seven dwellings, selected for the presence of an environment propitious to sandflies. Lutzomyia longipalpis predominated (94.3%), followed by Nyssomyia neivai (3.8%), Evandromyia lenti, Ev. cortellezzi and Brumptomyia brumpti. A total of 4,115 specimens of Lu. longipalpis were captured with a 3:1 male/female ratio. The highest average of sandflies/night/trap was obtained in the rainy season (NovemberMarch) and the highest frequencies of males and females were observed in March 2013. A positive and significant association was detected between the number of sandflies/trap/night and the total monthly rainfall (Spearman r= 0.48). Vector density constitutes the main parameter related to the dynamic transmission of vector borne diseases and the knowledge of the monthly distribution of the vector could contribute to the design and planning of control measures as well as to other strategies for the prevention of the disease. Our results suggest that the risk of transmission of L. infantum chagasi in Panorama city increases in the rainy season and that vector control activities should be undertaken especially in that period. Financial support: FAPESP – Process no. 2012/03751-4 and no. 2012/01442-4 N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 71-P Ecology & Epidemiology ID 72-P Ecology & Epidemiology FREQUENCY OF LUTZOMYIA LONGIPALPIS IN HENHOUSES IN AN AMERICAN VISCERAL LEISHMANIASIS ENDEMIC AREA OF SÃO PAULO STATE MICROPYGOMYIA QUINQUEFER (DYAR, 1929) (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE): A PUCTATIVE VECTOR OF LEISHMANIA SPP.? 1 2 2 Fredy Galvis Ovallos , Anaiá da Paixão Sevá , Estela Galucci , Rodrigo Martins Soares3, Fernando Ferreira3, Eunice Aparecida Bianchi Galati4 1. Postgraduate Program in Public Health, School of Public Health, University of São Paulo- USP, Brazil. 2.Postgraduate Program in Experimental Epidemiology of Zoonoses, School of Veterinary Medicine and Animal Science, University of São PauloUSP, Brazil. 3.Department of Preventive Veterinary Medicine and Animal Health, School of Veterinary Medicine and Animal Science, University of São Paulo-USP, Brazil. 4.Department of Epidemiology, School of Public Health, University of São Paulo – USP, São Paulo, SP, Brazil [email protected] 95 Cristian Ferreira Souza1; Paula Dias Bevilacqua2; José Dilermando Andrade Filho3; Reginaldo Peçanha Brazil4; Patrícia Flávia Quaresma5 1 Programa de Pós-Graduação em Biologia Parasitária- Instituto Oswaldo Cruz Fundação Oswaldo Cruz–Manguinhos, Rio de Janeiro–RJ. Brazil. 2Laboratório de Leishmanioses-Centro de Pesquisas René Rachou/Fiocruz, Belo Horizonte–MG. Brazil. 3Centro de Referência Nacional e Internacional para Flebotomíneos, Centro de Pesquisas René Rachou/Fiocruz, Belo Horizonte–MG. Brazil.) 4 Laboratório de Doenças Parasitarias-Instituto Oswaldo Cruz - Fundação Oswaldo Cruz–Manguinhos-Rio de Janeiro–RJ. Brazil. 5Departamento de Veterinária Universidade Federal de Viçosa-Campus Universitário-Viçosa–MG. Brazil [email protected] Keywords: Henhouses, Visceral leishmaniasis, Panorama-SP American visceral leishmaniasis (AVL), caused by Leishmania infantum chagasi and transmitted mainly by Lutzomyia longipalpis, is widespread in Brazil and in recent decades has expanded to new regions of the country. This study aimed to describe the effect of the removal of feces from henhouses on the density of AVL in São Paulo State. The sandfly captures were made in eight dwellings: three with henhouses from which the chicken feces were removed (FR+), three with henhouses without the removal of the chicken feces (FR-) and two residences without henhouses (Control - C). The captures were undertaken with two automatic light traps in each residence between July 2012 and June 2013. A total of 2,238 males and 746 females of Lu. longipalpis were captured. 76.6% of the insects were captured in the FR- houses, followed by C (14.3%) and FR+ (9.0%). Our results suggest that the removal of the feces from henhouses may have contributed to the decreases in Lu. longipalpis frequency, because of the removal of the organic matter necessary for the establishment of breeding sites. However, this result should be carefully analyzed because the sandfly frequency is also affected by other environmental characteristics such as vegetation cover and the availability of hosts. Henhouses are recognized as risk factors for AVL transmission, nevertheless the possibility of their elimination from urban areas constitutes a challenge in view of the fact that their presence is associated with socioeconomic and cultural factors. Further studies should be undertaken covering a larger sample of households with henhouses so as better to assess the effect of feces on the frequency of this sandfly species. Financial Support: FAPESP - Process no. 2012/03751-4 and no 2012/01442-4. Keywords: Leishmaniasis, sand fly, trypanosomatid, natural infection Micropygomyia quinquefer is a species of sand flies which, up to now, is not suspected of participating in the transmission of any Leishmania cycle. This is mainly due to the fact the species integrate a group of sand flies, which according to the literature, feeds exclusively on cold-blooded animals. This study aimed to verify the presence of natural infection in females of Mi. quinquefer captured in the urban area of the municipality of Timoteo, Minas Gerais, Brazil. Sand fly collections were performed in the city with Falcao light traps, exposed monthly for two consecutive nights during the period of November/2009 to October/2010. Captured females who had no signs of blood meal in their abdomen, were separated into pools of up to 10 females of the same species and used in molecular assays for the detection of natural infection by Leishmania spp. by PCR and RFLP ITS1. 1,958 sandflies were collected, with Nyssomyiawhitmani the most captured species (66.5%) and Mi. quinquefer the second (15.4%). Among the sand fly species of Mi. quinquefer, 251 (83.1%) females were divided into 91 pools for molecular analysis, 23 (25.3%) were positive for Leishmania spp. Infection by PCR of ITS1 and 8 (8.8%) were positive for L. braziliensis by RFLP of ITS1. This is the third record of Mi. quinquefer with Leishmania DNA in Brazil. This finding is of particular ecological significance; since it is believed that females of Mi. quinquefer feed on cold-blooded animals and cannot has a role in the transmission cycle of Leishmania sp. Another interesting detail was the identification of a large number of positives pools for L. braziliensis. Hence that L. braziliensis only infects mammalian species, these results suggest that Mi. quinquefer can feed on warm-blooded animals, or even that molecular markers used are amplifying DNA fragment from other trypanosomatid, not yet described by science. Although it cannot establish this species as a vector of L. braziliensis, we cannot ignore that it was the second most frequently captured species in the study area, being characterized by the occurrence of human cases. This shows the necessity of other studies to understand the participation of this sand-fly species as a vector in eco-pidemiology of leishmaniasis in the area. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 73-P Ecology & Epidemiology ID 74-P Ecology & Epidemiology CANINE ANTIBODY RESPONSE TO PHLEBOTOMUS PERNICIOSUS AND PHLEBOTOMUS ORIENTALIS BITES. EPIDEMIOLOGICAL ASPECTS OF VECTOR, PARASITE, AND DOMESTIC RESERVOIR IN AREA WITH NO REPORTED HUMAN CASES OF VISCERAL LEISHMANIASIS IN BRAZIL Tereza Kratochvílová1, Tatiana Košťálová1, Michaela Kindlová1, Petra Sumová1, Michal Šíma1, Iva Rohoušová1, Marina Gramiccia2, Luigi Gradoni2, Asrat Hailu3, Gad Baneth4, Alon Warburg5, Petr Volf1 1 Department of Parasitology, Faculty of Science, Charles University in Prague, Vinicna 7, 128 44 Prague 2, Czech Republic; 2 Unit of Vector-borne Diseases and International Health, MIPI Department, Istituto, Superiore di Sanità, Rome, Italy; 3 Department of Microbiology, Immunology & Parasitology, Faculty of Medicine, Addis Ababa University, P.O. Box 9086, Addis Ababa, Ethiopia; 4 School of Veterinary Medicine, Hebrew University, P.O. Box 12, Rehovot 76100, Israel; 5 Department of Parasitology, The Kuvin Centre for the Study of Infectious and Tropical Diseases, Hadassah Medical School, The Hebrew University of Jerusalem, Jerusalem, Israel. 96 Fabiana de Oliveira Lara-Silva1, Érika Monteiro Michalsky1, Consuelo Latorre Fortes-Dias2, Vanessa de Oliveira Pires Fiuza3, José Eduardo Marques Pessanha3, Shara Regina da Silva1, Daniel Moreira de Avelar1, Maiara Alves Silva1, Ana Cristina Vianna Mariano da Rocha Lima1, Ailton Junior Antunes da Costa1, George Luiz Lins Machado-Coelho4, Edelberto Santos Dias1 1 Laboratório de Leishmanioses, Centro de Pesquisas René Rachou/Fiocruz, Belo Horizonte, Minas Gerais, Brazil; 2Diretoria de Pesquisa e Desenvolvimento, Fundação Ezequiel Dias, Belo Horizonte, Minas Gerais, Brazil; 3Secretaria Municipal de Saúde de Belo Horizonte, SMSA/PBH, Brazil; 4Universidade Federal de Ouro Preto, Minas Gerais, Brazil [email protected] [email protected] Keywords: P. perniciosus, P. orientalis, antibody, canine In the Mediterranean area, dogs serve as the main reservoir host of Leishmania infantum, the causative agent of zoonotic visceral leishmaniasis (VL). In East Africa, the role of dogs in the epidemiology of VL caused by L. donovani is not clear. In both areas, however, the disease is transmitted by sand flies of the subgenus Larroussius; Phlebotomus perniciosus and P. orientalis are major vectors of VL in the Mediterranean and East Africa, respectively. Frequent contact of the sand fly vector with the host is a prerequisite for transmission and therefore we were interested to learn more about exposure of dogs to these sand fly species. Sand fly saliva inoculated into the feeding site during bloodmeal intake elicits an antibody response that can be utilized as a specific marker of exposure. The intensity of exposure correlates with the antibody response against saliva and may serve as an epidemiological tool for estimating the vector bites and also for the risk of Leishmania transmission. However, since the use of whole salivary homogenate is limited by maintenance of sand fly colonies and laborious dissection of their salivary glands, recombinant salivary proteins have recently been tested to replace salivary gland homogenate in serological surveys. In our study we identified major salivary antigens of P. perniciosus and P. orientalis by western blotting and mass spectrometry (MALDI-TOF) and tested them as potential markers of dog exposure to these sand fly vectors. The two most promising P. perniciosus recombinant proteins, 43-kDa yellow-related protein (rSP03B) and 35.5-kDa apyrase (rSP01) were tested by ELISA with more than 750 canine sera collected in southern Italy, an area endemic for VL. Similarly, we studied reaction of P. orientalis salivary proteins with canine sera collected in a L. donovani focus in Ethiopia. Canine sera originating from dogs living in non-endemic countries were used for negative control. Experiments showed that dogs from both endemic areas had significantly increased IgG levels against saliva of the local vector. Moreover, the results proved that recombinant salivary proteins of sand flies can fully replace salivary gland homogenate in ELISA tests. Keywords: Phlebotominae, Lutzomyia longipalpis, Leishmania infantum, natural infection Visceral leishmaniasis (VL) is a neglected disease of great importance in Public Health. In the Americas, about 97% of the human cases of VL occur in Brazil. In the last few years, the disease has been expanding to medium and large size cities and the actions taken by the Brazilian Program for Surveillance and Control of VL are more intense where the epidemiological transmission risk (ETR) is higher. Belo Horizonte, the sixth most populous Brazilian city, is among the state capitals with intense ETR of VL. The ETR, however, varies depending on the urban district analyzed. We focused our twoyear study in districts of Belo Horizonte with no reported human cases of VL, so far, in an effort to draw a current epidemiological profile of the local situation concerning vector, parasite, and domestic reservoir. Lutzomyia longipalpis, the main vector of VL in Brazil, was highly abundant, mostly in the peri-domiciles. The species comprised 96.5% of all the phlebotomine sand flies captured, with increased numbers after rainfall peaks. DNA of Leishmania infantum, the etiological agent of VL, was detected in 88.5% of the Lu. longipalpis pool samples. Le. infantum DNA was also present in the cortelezzii complex and in Lutzomyia lloydi. DNA of Leishmania braziliensis, etiological agent of the cutaneous leishmaniasis, was also identified in Lu. longipalpis. Among the 1,410 dogs serologically tested for VL, 3.6% were positive. These dogs were additionally submitted to two parasite-based and one molecular-based diagnostic tests. The main result was that Leishmania infection was confirmed in all of them when the spleen tissue was used as DNA source in the molecular test. Moreover, the infecting parasite was genotyped as Le. infantum in all of them. We concluded that the parasite–vector and the parasite–domestic reservoir interfaces are very active in those districts, even in the absence of reported human cases of VL, therefore providing favorable conditions to local outbreaks in a short period of time. Financial support: FAPEMIG/FIOCRUZ/SMSBH. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 75-P Ecology & Epidemiology ID 76-P Ecology & Epidemiology SANDFLIES FAUNA SURVEY AND DNA DETECTION OF LEISHMANIA INFANTUM IN LUTZOMYIA LONGIPALPIS CAPTURED IN THE CITY OF SABARA, MINAS GERAIS, BRAZIL P H L E B O TO M I N E S A N D F L I E S ( D I P T E R A : PSYCHODIDAE) IN CUTANEOUS LEISHMANIASIS ENDEMIC AREAS OF THREE MUNICIPALITIES OF THE STATE OF PARÁ, BRAZIL Érika Monteiro Michalsky1, Josiane Valadão Lopes1, Fabiana Oliveira Lara E Silva1, Maiara Alves Silva1, Ana Cristina Vianna Mariano Da Rocha Lima1, João Carlos França-Silva2, Fabiana Rodrigues Da Paixão1, Ailton Junior Antunes Da Costa1 & Edelberto Santos Dias1 Ana de Nazaré Martins da Silva; Edna Aoba Yassui Ishikawa; Mônica Fadul; Thiago Vasconcelos dos Santos; Yago Costa Vasconcelos dos Santos; Patrick Thiago Carvalho de Araújo e Nathália Queiroz Santos 1 2 Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, Belo Horizonte, Brasil Universidade Federal de Minas Gerais, Belo Horizonte, Brasil 97 Núcleo de Medicina Tropical, Universidade Federal do Pará, Brazil [email protected] [email protected] Keywords: Lutzomyia, sandfly, cutaneous leishmaniasis, Brazil Keywords: Leishmania infantum, Lutzomyia longipalpis, natural infection, Sabará Visceral leishmaniasis (VL) is a neglected disease of great importance in Public Health, which is present in 65 countries around the world. In Brazil, VL urbanization is a fact, with increasing numbers of human cases and high prevalence of the canine cases in large size cities. In the New World, Leishmania infantum is the etiologic agent and its transmission occurs through the bite of the females of Lutzomyia genus sandflies. LV is distributed in the city of Sabara, characterizing the region as an endemic area. The aim of this work was to survey the city's sandflies fauna, as well as study the behavior of the species in domestic environments and detect natural infection of Lutzomyia longipalpis by Leishmania spp. The entomological captures were performed in 8 districts of the city (Alvorada, Novo Alvorada, Alvorada Velho, Bom Retiro, Ana Lúcia, Rio Negro, Nova Vista e Casa Branca), during the period of 01/2011 to 12/2012. These were made monthly for three consecutive days using the HP type light traps installed inside and outside homes. The males collected were stored in 70% ethanol for taxonomic identification and females were dissected and analyzed by Nested /PCR, afterwards, DNA sequencing was made for identification of circulating Leishmania species. 519 specimens were captured and 4 different species were identified. The species distribution was: L. longipalpis (95.0%), L. cortelezzii (2.3%), L. whitmani (1.5%) and L. intermedia (0.2%). Regarding the behavior of the species, 13% were caught inside and 87% outside the homes. Of the 9 L. longipalpis "pools" analyzed, the 353 bp fragment typical of Leishmania spp. was observed in 3 of them, keeping the minimum infection rate around 33%. Of the 3 female L. longipalpis "pools" positive for Leishmania, were made DNA sequencing and sequences alignment and we observed that the Leishmania species circulating in the vectors of city of Sabara is L. infantum. The sandflies fauna showed leishmaniasis vector species, both visceral and cutaneous. The large number of captured L. longipalpis (95%) and a high infection rate by L. infantum (33%), associated with high canine positivity (4.25%) and the occurrence of human cases of VL, demonstrate that the cycle of the disease transmission is well established in the studied urban area. Financial support: FAPEMIG/FIOCRUZ/CNPq/SMS-Sabará The number of cutaneous leishmaniasis cases (CL) notified from 2010 to 2012 has demonstrated a fast expansion in state of Pará, Brazil, and the Healthy Department rated 26.57% of the municipalities as very high transmission area. The aim of this work was to study the fauna and to detect natural infection by Leishmania spp in sand flies captured in six communities with or without recent record of CL in human: Villages Betel and Paraíso (Dom Elizeu); Settlements Rossi Gabriel and Nova Vida (Ulianópolis); Villages Água Branca and Santa Helena (Rondon do Pará). Sand flies were captured using CDC light traps placed in peridomicile areas. The traps were installed 1.5 m above ground level at 6:00 pm and removed the next day at 6:00 am. The taxonomic identification of each specimen determined was based on Young and Duncan (1994) and all females were selected for PCR analysis. In 2011, a total of 229 specimens were captured: Dom Elizeu: Lutzomyia flaviscutellata (35.21%), Lu. evandroi (18.31%), Lu. whitmani (11.27%), Lu. choti (9.86%), Lu. davisi (9.86%), Lu. sericea (1.41%), Lu. termithophila (1.41%), Lu. bacula (1.41%), Lu. sordellii (1.41%), Lu. hirsusta hirsuta (1.41%); Rondon do Pará: Lu. evandroi (60.0%), Lu. triacantha (13.34%), Lu. shannoni (6.67%), Lu. gomezi (6.67%), Lu. dasypodogeton (3.33%), Lu. evandroi (3.33%), Lu. spinosa (3.33%), Lu. trinidadensis (3.33%), and Lu. whitmani (3.33%); Ulianópolis: Lu. triacantha (42.19%), Lu. hermanlenti/dreisbachi (23.47%), Lu. evandroi (9.37%), Lu. wellcomei/complexa (7.037%), Lu. flaviscutellata (5.47%), Lu. umbratilis (2.34%), Lu. whitmani (1.56%), Lu. gomezi (0.78%), Lu. longipalpis (0.78%), Lu. shannoni (0.78%), Lu. trinidadensis (0.78%), Lu. ubiquitalis (0.78%), Lu. davisi (0.78%). None of the female sand flies were found infected by Leishmania spp. Despite the low number of specimens captured, Lu. flaviscutellata, vector of L. (L.) amazonensis was the predominant species in Dom Elizeu. In Ulianópolis, among medical interest species, Lu. wellcomei/complexa the main vector of L.(V.) braziliensis in the region, was the most prevalent, followed by Lu. flaviscutellata. The presence of sand fly fauna as recorded demonstrates the need for epidemiological surveillance of CL in these human settlements, many of them located near the forest areas. Supported by MS/CNPq/FAPESPA and SESPA. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 77-P Ecology & Epidemiology ID 78-P Ecology & Epidemiololgy PRELIMINARY STUDY OF SANDFLIES CAPTURED IN THE CITY OF MACEIÓ-ALAGOAS-BRAZIL EDUCATION: DEMOGRAPHIC AND SOCIAL IMPACTS ON THE RATE OF INCIDENCE OF VISCERAL LEISHMANIASIS IN THE STATE OF TOCANTINS, BRAZIL (2010) Ana Lúcia Moreira Macena, Cláudia Cavalcante De Matos Rodarte, Maria Elisabel Tavares Da Silva, Carlos Fernando Rocha Dos Santos, Magliones Carneiro Lima, Grimoaldo Braga Da Rocha Neto 98 Secretaria de Saúde Maceió-Alagoas-Brasil Ana Flávia de Morais Oliveira, Hebert Lima Batista, Keily Cristiny Azevedo Leite, Helio Márcio Nunes Lacerda [email protected] Instituto Federal de Educação, Ciência e Tecnologia do Tocantins [email protected] Keywords: Sandflies, Maceió, Captured Keywords: leishmaniasis, social indicators, incidence American visceral leishmaniasis is an anthropozoonosis whose etiologic agent is the protozoan Leishmaniachagasi. Transmission occurs through the bite of the infected female sandfly Lutzomyialongipalpis, which has been registered both in natural ecotopes as in rural and urban environments, next to domestic animals and human habitations (Roach et al. 2004). In Latin America, visceral leishmaniasis has been described in 12 countries, with 90% of the cases occurring in Brazil, especially in the Northeast Region (MS/2006). Maceio is a city known as endemic for visceral leishmaniasis, the Center for Zoonosis Control of the city develops routinely entomological investigations aiming to gather both quantitative and qualitative information about sandflies. To the execution of the leishmaniasis control program, the sand flies were captured in a non-selective way by CDC light traps. The sampling was done where the occurrence of human and/or canines positive cases were registered, being previously installed around 20 traps per night, 1.5m above ground level, for a period of 12 hours. In the urban areas, the traps were installed intra and extra household, in a sequence of two nights; while in the rural areas the same trap distribution and sampling period were used but an extra trap was added outside the house. In the year of 2013, 1.125 sandfly specimens were found in 13 locations in the city of Maceió. Of these, 246 were identified as Lutzomyialongipalpis; 6 L.migonei; 721 L.whitmani; 31 L.corumbaensis; 04 L.quinquefer; 56 L.chotti; 05 L.sallezi; 01 L.tupinambay; 45 L.intermedia; 03 L.evandroi; 04 L.sp (Pressatia); and 03 L.spp (lenti/carmelinoi). According to the Ministry of Health (2006), two species are related to the transmission of the disease in Brazil: L. longipalpis and L. cruzi. Visceral leishmaniasis is recognized by the World Health Organization as one of the most endemic and lethal diseases in the world, not just because of its deadliness, but also because it is very difficult to treat. This study aimed to analyze the relationship between the number of cases of VL and the rate of illiteracy, and the average per capita income. In addition to evaluating the relationship between the disease and the number of inhabitants, this study also sought to describe the incidence of the disease in the state of Tocantins during 2010. A descriptive, analytical and retrospective study was done in 2010, using secondary data from IBGE (demographical and social data) and DATASUS (death rate) through a sample of positive cases of VL among Tocantins residents. Microsoft Excel 2010, Tabwin, and Biostat were used to process the data. The general incidence of this disease was measured from a sample of 100 thousand inhabitants, taking into consideration only the cases confirmed by the laboratorial and clinic/epidemiologic criteria. The population used to generate these statistics came from the 2010 Demographical Census. These social indicators were pulled from the IBGE (Instituto Brasileiro de Geografia e Estatística). The social indicators in the IBGE data was taken from the median per capita income and the average illiteracy rate. The Pearson theory was applied in order to understand the relationship between VL incidence, social variables, and the number of inhabitants. The statistic was considered significant when the value of p was less than 0.05. The study was made exclusively with secondary data. The research demonstrated that the median per capita income pulled from 89.2% of the municipalities in Tocantins was less than R$ 500.00 per month. The illiteracy rate was considered alarming: when 42 (30.2%) of the municipalities showed an illiteracy rate greater than 20%. Approximately 74 (53.2%) of the municipalities had a population smaller than five hundred thousand inhabitants. The majority of LV outbreaks were concentrated within 7 municipalities that affected 38% of the state's total population. The incidence rate of VL was higher than 4 out of 100,000 inhabitants in 41% of the municipalities. In conclusion, there was a significant correlation between the incidence of VL, the state's illiteracy rate, and the state's median per capita income. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 99 ID 79-P Ecology & Epidemiology ID 80-P Ecology & Epidemiology ECO-EPIDEMIOLOGY OF LEISHMANIASIS IN THE REGION OF SERRA DO CIPO, A MAJOR TOURISTIC CENTRE OF MINAS GERAIS PHLEBOTOMINE SAND FLY FAUNA (DIPTERA: PSYCHODIDADE) FROM THE LEISHMANISIS ENDEMIC FOCI IN MATO GROSSO, BRAZIL Rosana Silva Lana, Erika Monteiro Michalsky, João Carlos França Silva, Fabiana de Oliveira Lara e Silva, Consuelo Latorre Fortes Dias, Juliana Martins & Edelberto S. Dias Sinara Cristina de Moraes, Alan Lane Melo e Elizabeth Ferreira Rangel Depto. de Parasitologia/ICB/UFMG, Belo Horizonte- MG Fundação Oswaldo Cruz - Centro de Pesquisas René Rachou [email protected] [email protected] Keywords: Leishmaniasis, sandflies and Leishmania Keywords: Leishmaniasis, Leishmania, Epidemioly The purpose of this study was to evaluate the transmission of leishmaniasis in Jaboticatubas, Serra do Cipó, tourist region of Minas Gerais. For this, entomological studies and urban reservoirs of the disease were conducted. For entomological studies, eight districts were selected, taking into account the occurrence of human cases, and light traps HP type were placed in the outdoors in 10 homes for three consecutive nights per month from May 2012 to April 2013. The sand flies studied consisted of 3,104 specimens , quite diverse with 17 species of Lutzomyia and two females of the genus Brumptomyia. With regard to species of medical importance and also numerous, emphasize the meeting of Lu.whitmani (33.80 %), Lu. intermedia (14.60%) and Lu. longipalpis (7.96 %). The general rate of natural infection of sand flies by Leishmania was 3.40%. 249 "pools" of different sand fly species, among them 32 were positive. To study the rate of canine infection, census survey among domestic dogs in urban area was carried out. 1,105 serum samples were tested by the DPP® and confirmed positive infections by ELISA immunoenzymatic assay, obtaining the average prevalence of canine 7.78%. Of the 86 positive dogs, a random sample of 11 was selected , which were necropsied and samples of biopsy tissue obtained were performed " imprints ", culture of parasites and PCR. Georeferencing was conducted in urban areas of all canine and human leishmaniasis cases in the last 6 years, and built up a map showing the distribution of cases. The PCR of marrow aspirate showed a higher positivity (63.60%) compared to skin (9.00%). While the "imprints", the positivity rate of skin was 36.36 % and 0 % marrow. As for the culture of marrow aspirates, the positivity was 27.27%. Our results indicate the urbanization of leishmaniasis in the town due to high vector density and infection by Leishmania, expressive average rate of prevalence canine infection and recent records of human cases. Therefore, is important to intensify control measures in order to prevent the problem from getting worse in the town. The Leishmaniases are a group of diseases caused by the protozoa of the genus Leishmania (Kinetoplastida:Trypanosomatidae) which affect several orders of mammals, including humans, and are transmitted by the bite of some species of sandflies (Diptera:Psychodidae). The Leishmaniasis in Brazil has shown a large geographic expansion. The aim of this study was to compare the indoor sand flies, peridomicile with that of outside the home (sites near forests) by entomological research in the urban area, the extra urban (adjacent to urban) and wild, in the county of Barra do Garças, Mato Grosso. The entomological survey was carried out from July 2013 to February 2014 with the use of CDC light traps, monthly collections were made from 3 consecutive nights around 20 houses, of which 8 captured were inside. Catches of sandf lies were also held outside the house and in the wild, one night per month, by trapping Shannon light traps. The sand fly fauna comprises 32 species representing two genera (Lutzomyia and Brumtomyia)from 4,181 specimens captured. The most frequent species were Lutzomyia (L.) longipalpis (46.5%), followed by L. lenti (15.1%), L. (N.) whitmani (12%), L. carmelinoi (9%), L. (L.) cruzi (7.4%), and other species were twenty seven (10 %) as L. evandroi, L. acanthopharynx, L (S.) sordellii, B. brumpti, L. termitophila, L. hermanlenti, L. goiana, L (P.) davisi, L. longipennis, L. (E.) teratodes, L. (P.) punstigeniculata, L. oliveirai, L. (P.) christenseni, L.(P.) lutziana, L. brasiliensis, L. ferreirana, L. peresi, L. walkeri, L. (N.) antunesi, L. aragaoi, L. (N.) flaviscutellata, L. (P.) shannoni, L. baculus, L. cerradicula, L. complexa, L. sallesi e L. saulensis. For peridomicile were collected 32 species totaling 3,630 sand flies (87.8%), in house indoor were collected 13 species, totaling 343 sand flies (8.2%), outside the home 14 species, totaling 141 sand flies (3.4%) and in the wild area 7 species, totaling 27 sand flies (0.6%). The predominant species caught from these areas was L. (L.) longipalpis, with 57% of the total species collected inside the home, 45.2 % and 50.3 % peridomiciles outside the home. The exception was the wild area with 29.5% of L. (L.) longipalpis, the specimens that match with other species represents 70.4 %. It is suggested that the transmission cycle of AVL in the urban area is apparently frequented by the sand fly vector associated with the presence of autochthonous cases. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 81-P Ecology & Epidemiology ID 82-P Ecology & Epidemiology SAND FLY FAUNA (DIPTERA: PSYCHODIDAE) IN THE ENDEMIC AREA OF LEISHMANIASIS IN BARRA D O G A R Ç A S – M ATO G R O S S O A N D T H E DEMOGRAPHIC INFORMATION ON LUTZOMYIA (LUTZOMYIA) LONGIPALPIS AND LUTZOMYIA (NYSSOMYIA) WHITMANI THE ABUNDANCE AND DISTRIBUTION OF PHLEBOTOMINE SAND FLY IN SICILY Torina A.1,2; Reale S.1, 2; Blanda M.1; La Russa F.1; Intile S.1; Lelli R.1; Vitale F.1 1 Istituto Zooprofilattico Sperimentale della Sicilia, Palermo, Italy; University of Messina, Italy. Sinara Cristina de Moraes, Lucas Heber Mariano dos Santos, Thiago Nunes Alexandre, Thais Brito do Nascimento 100 2 Vet. Sc. Dep. [email protected] Depto. de Parasitologia/ICB/UFMG, Belo Horizonte- MG [email protected] Keywords: Leishmaniasis, sandflies, Leishmania Sand flies are transmitting a number of diseases, including leishmaniasis. The objective of the study was to assess the sand fly fauna present in the neighbourhoods of Barra do Garças - MT and its associate richness and population density with demographic data; to determine the neighbourhoods with the highest risk of transmission of leishmaniasis. Sand flies were captured from 2013 using CDC light traps. The light traps were used in peridomestic areas near animal shelters for three consecutive nights from 06:00 pm-06:00 am of 41 neighbourhoods. The captured specimens were fixed in 70% alcohol and identified in accordance with Young & Duncan. Demographic data were collected from localities system - SISLOC. The diverse phlebotomine fauna identified from 1650 specimens include 31 species under 2 genera, Shannon diversity H = 2.619, these were the most abundant species Lutzomyia (Nyssomyia) whitmani (48.42%) , followed by L.(Lutzomyia) longipalpis (16.84%) , L.(L.) cruzi (10.9%), L. lenti (6.96%) and a further twenty seven (<3%) as B. pintoi, Brumptomyia brumpti, L. abunaensis , L. aconthopharynx, L. aragaoi L. brasiliensis, L. carmelinoi, L. carrerai carrerai, L. cerradincula, L. christenseni, L. davisi, L. evandroi, L. ferreirana, L. flaviscutellata, L. goiana, L. hermanlenti , L. longipennis, L. lutziana, L. oliveirai, L. punctigeniculata, L. runoides, L. sallesi, L. saulensis, L. sordellii , L. teratodes, L. termitophila e L. walkeri. The neighbourhoods with the highest densities of the species L. (L.) longipalpis and L. (N.) whitmani were: DERMAT with 9.3% and 57.35%, São José with 37.80% and 13.62%, Palmares with 22.15% and 7.16%, Vila Maria 11.26% and 0.35% and Novo Horizonte with 2.63% and 10%; corresponding to 14.5% inhabitants. The urban area have plots free and some neighbourhoods make border with conservation area Serra Azul Park State. It's common the presence of chickens. The sandfly fauna in Barra do Garças is varied and some species identified are considered to be potentials vectors of American cutaneous leishmaniasis (ACL) in Brazil, as L. flaviscutellata, L. wellcomei and L. (N.) whitmani, or vectors of American visceral leishmaniasis (AVL), as L. (L.) longipalpis, L. (L.) cruzi. The neighbourhood's studied, 38 of them had at least one related species transmission of leishmaniasis. The data suggest that the cycle of leishmaniasis in the region is highly urbanized, as the wide distribution of vectors in the urban area. Keywords: Epidemiology, Phlebotomus, Sergentomya Sand flies (Psychodidae: Phlebotominae), the vectors of Leishmania as well as of phleboviruses, have been known in Mediterranean for long time. In this paper the diversity and the abundance of phlebotomine vectors were investigated in Sicily through the 2012 sand fly season using Onderstepoort-type backlight traps. Traps were set up from 1 h before sunset to 1 h after sunrise and collected the next morning. The largest number of sand flies captured in one night was 11396 in the capture of 9th, July, while the lowest number was 1 in catches of April and December. June and July were the month with the highest abundance (n.of insect/n. of positive catches), in the other months there was a progressive decrease. P. perniciosus only had a trend characterized by two peaks, one in June and the second slight increase in August, P. mascittii had a peak in June, while the other species [what are they] showed a peak in July. A total of 101123 sand flies was identified as belonging to 7 species: Phlebotomus perfiliewi (40.31%), P. perniciosus (24.84%), P. neglectus (16.67%), P. mascittii (3.31%), P. papatasi (0.20%), P. ariasi (0,05 %) and Sergentomyia minuta (14.62%). The two species considered as the main vectors of human leishmaniasis in Italy, P. perniciosus and P. perfiliewi, were the most abundant, although the higher prevalence of P. perfiliwi can be explained by the fact that the catch was made in rural areas and P. perniciosus is the main anthropophilic vector, while P. perfiliewi is considered mainly zoophilic. Moreover, P. neglectus is a proven vector of L. infantum in Greece. In conclusion the risk of transmission of Leishmania in Sicily remains high and competent vectors are well represented and it can be seen a prolonged season of sand flies and an increase of the population in terms of diversity of identified species, abundance and prevalence. Acknowledgements We thank Rosa Filippi for the helpful and precious technical support and Gabriella Serafino Agrusa. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 83-P Ecology & Epidemiology ID 84-P Ecology & Epidemiology LUTZOMYIA LONGIPALPIS DISTRIBUTION AND ABUNDANCE IN THE CITY OF CORRIENTES, NORTHEASTERN ARGENTINA A STUDY OF PHLEBOTOMINAE IN MODIFIED WHITE AND BLACK SHANNON TRAPS IN A MAROON AREA AT PIRAPUTANGA DISTRICT, AQUIDAUNA, MATO GROSSO DO SUL, BRAZIL Berrozpe Pablo, Araujo Analía, Santini Maria Soledad, Utges Maria Eugenia; Salomon Oscar Daniel REDILA- Instituto Nacional de Medicina Tropical; Centro Nacional de Diagnostico e Investigación en Endemoepidemias; CONICET [email protected] Keywords: Lu. longipalpis, environmental stratification. Corrientes was sampled in two stages during summer. In the first stage (December 2013), the city was divided in areas of 800 x 800 m to evaluate Lutzomyia longipalpis distribution in the whole city. In the second stage (February-March 2014) we evaluated the distribution of Lu.longipalpis abundance according to environmental strata. A total of 6 strata were discriminated based on satellite images 'Spot 5' HRG1. In each stratum we selected at least three positive sites for Lu. longipalpis (first stage samples). In each peridomestic environment selected under 'worst case' criteria, mini-traps of UV-light were located during 5 consecutive nights from 5 p.m. to 9a.m. (first stage: 51 traps/night; second stage: 21 traps/night. At each peridomicile we recorded maximum and minimum temperature and relative humidity, and the georeference. The specimens were clarified with lacto-phenol and identified according to Galati (2003). A total of 1390 sandflies were captured (555 in the first stage, and 835 in the second stage). In the first stage a total of 5 species were captured: Lu. longipalpis (58.25%), Nyssomyia neivai (31.75%), Mygonemyia migonei (7.40%), Evandromyia cortelezzii complex (1.0%) and Mycropigomyia quinquefer (1.6%). In the second stage 4 species were captured: Lu. longipalpis (94.3%), Ny. neivai (2.7%), Mg. migonei(2.3%), Ev. cortelezzii complex (0.6%). These preliminary results showed that Lu. longipalpis is the most abundant specie in the city of Corrientes. When we analyzed the environmental strata, we could describe a decreasing gradient of abundance from primary vegetation towards the most densely inhabited center of the city. We also found areas of high (>30 Lu. longipalpis individuals/trap/night), medium (15 to 30 Lu. longipalpis individuals/trap/night) and low abundance (< 15 Lu. longipalpis individuals/trap/night) in sites associated to primary riparian vegetation of the River Paraná. Areas whithin medium and low abundance of Lu.longipalpis in peripheral neighborhoods and finally areas of low abundance of Lu. longipalpis in Downtown. 101 Jucelei de Oliveira Moura Infran1,4; Daiana Alovisi Souza1; Wagner Fernandes de Souza1; Aline Etelvina Casaril2; Gabriel Utida Eguchi3; Elisa Teruya Oshiro4; Carlos Eurico Fernandes1; Alessandra Gutierrez de Oliveira1,2,4 1 .Curso de Pós-graduação em Biologia Animal - Centro de Ciências Biológicas e da Saúde/Universidade Federal de Mato Grosso do Sul, 2.Curso de Pós-graduação em Doenças Infecciosas e Parasitárias- Faculdade de Medicina/ Universidade Federal de Mato Grosso do Sul; 3.Faculdade de Medicina Veterinária e Zootecnia Universidade Federal de Mato Grosso do Sul; 4.Laboratório de Parasitologia-Centro de Ciências Biológicas e da Saúde/Universidade Federal de Mato Grosso do Sul Cidade Universitária, 79070-900, Campo Grande, MS, Brazil. [email protected] Keywords: sand flies, Shannon trap, anthropophily The study was carried out at Furnas dos Baianos, a Maroon area located at the district of Piraputanga in Aquidauana, Mato Grosso do Sul-MS, Brazil. From September 2013 to June 2014, uninterrupted captures were performed for 24h on a monthly basis in modified white and black Shannon traps placed at the foot of the mountain. Temperature and relative humidity were recorded during this period. A total of 707 sand flies was captured, 269 (39.03% males and 60.96% females) in white Shannon trap and 438 (62.32% males and 37. 67% females) in black shannon trap. 11 species were identified as: Brumptomyia avellari, Evandromyia lenti, Evandromyia saulensis, Lutzomyia díspar, Lu. longipalpis, Martinsmyia oliveirai, Micropygomyia peresi, Nyssomyia whitmani, Pintomyia misionensis, Psathyromyia bigeniculata. Lu. longipalpis was the most frequently found in both colour traps, followed by Ny. whitmani. The third most prevalent species was Pi. misionensis in the white Shannon trap and Lu. dispar in the black Shannon trap. Along the captures the researches realized they were attracting many sand flies and they were identified as Lu. longipalpis, Lu. dispar and Pi. misionensis. Interestingly many males (80.85%) of Lu. longipalpis were among them. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 85-P Ecology & Epidemiology ID 86-P Ecology & Epidemiology NYCTHEMERAL RHYTHM OF PHLEBOTOMINAE IN A MAROON AREA AT PIRAPUTANGA DISTRICT, AQUIDAUNA, MATO GROSSO DO SUL, BRAZIL SAND FLY FAUNA IN CHAPARE, BOLIVIA: AN ENDEMIC FOCUS OF LEISHMANIA (VIANNIA) BRAZILIENSIS Jucelei de Oliveira Moura Infran1,4; Daiana Alovisi Souza1; Wagner Fernandes de Souza1; Aline Etelvina Casaril2; Gabriel Utida Eguchi3; Elisa Teruya Oshiro4; Carlos Eurico Fernandes1; Alessandra Gutierrez de Oliveira 1,2,4 Marinely Bustamante Gomez1, Mery Diaz1, Jorge Espinoza1, Rudy Parrado1, Richard Reithinger2, 3, and Ana Lineth Garciá1 1 . Curso de Pós-graduação em Biologia Animal - Centro de Ciências Biológicas e da Saúde/Universidade Federal de Mato Grosso do Sul, 2. Curso de Pós-graduação em Doenças Infecciosas e Parasitárias- Faculdade de Medicina/ Universidade Federal de Mato Grosso do Sul; 3. Faculdade de Medicina Veterinária e Zootecnia - Universidade Federal de Mato Grosso do Sul; 4. Laboratório de Parasitologia-Centro de Ciências Biológicas e da Saúde/Universidade Federal de Mato Grosso do Sul Cidade Universitária, 79070-900, Campo Grande, MS, Brazil. [email protected] 102 1 IIBISMED, Facultad de Medicina, Universidad Mayor de San Simón, Cochabamba, Bolivia; 2Faculty of Infectious & Tropical Diseases, London School of Hygiene & Tropical Medicine, Keppel Street, London WC1E 7HT, United Kingdom; 3George Washington University School of Medicine and Health Sciences, Washington, D.C. 20035. [email protected] Keywords: leishmaniasis, Lutzomyia, Leishmania (Viannia), sand fly Keywords: sand flies, hourly activity, seasonality The study was carried out at Furnas dos Baianos, a Maroon area located at the district of Piraputanga, in Aquidauana, Mato Grosso do Sul (MS), Brazil. From July 2012 to June 2014, uninterrupted captures in modified Shannon trap placed at the foot of the mountain were performed for 24h on a monthly basis for two years. Temperature and relative humidity were recorded during this period. A total of 1.815 sand flies were captured and the following species were identified: Nyssomyia whitmani, Lutzomyia dispar, Lutzomyia longipalpis, Pintomyia misionensis, Psathyromyia bigeniculata, Evandromyia lenti, Micropygomyia oswaldoi, Brumptomyia avellari, Micropygomyia peresi, Martinsmyia oliveirai, Evandromyia saulensis, Pintomyia pessoai, Psychodopygus davisi, Evandromyia evandroi and Psathyromyia campograndensis. Nyssomyia whitmani (28.4%, 515 ∕ 1,815, with 51.0% of females and 49.0% of males) and Lu. longipalpis (20.4%, 371 ∕ 1,815, with 25.6 % of females and 74.4% of males) were the first and third most prevalent species, also they showed highest abundance of insects captured in dry season, being the activity time between 6pm - 5am and 7pm - 5am, respectively. These two species deserve attention because they are considered main vectors of Leishmania (Viannia) braziliensis and Leishmania (Leishmania) infantum in many regions of Brazil including Mato Grosso do Sul. Lu. dispar, (24.7%, 455 ∕ 1,815, with 98.5% of females and 1.5% of males), the second most prevalent species was found high activity in rainy season, between 6pm and 2am. Data on the distribution and abundance of Lutzomyia spp. (Diptera: Psychodidae) in Bolivia is scarce. Sand flies from an area of Leishmania (Viannia) braziliensis endemicity in the Isiboro-Secure National Park in the Department of Cochabamba were captured and identified to species. In total, 945 sand flies (789 females and 156 males) belonging to 15 species were collected from the four collection points in two study villages in 2007. With 549 (58.1%) specimens, Lutzomyia shawi was the most abundant species, followed by Lutzomyia (Trichophoromyia) sp. (22.2%), Lutzomyia llanosmartinsi (8.3%), Lutzomyia antunesi (4.3%), and Lutzomyia olmeca (2.1%). Abundance and species composition varied between rainy and dry seasons, with 99.3% of all sand flies being collected outdoors. Because of species abundance and confirmed Leishmania infection in previous entomological collections, we believe Lu. shawi is the vector of L. (Viannia) braziliensis in Isiboro-Secure National Park. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 87-P Ecology & Epidemiology ID 88-P Ecology & Epidemiology DIVERSITY AND NYCTHEMERAL RHYTHM OF PHLEBOTOMINES IN SAN JULIAN COMMUNITY (COCHABAMBA, BOLIVIA) STUDIES ON THE FAUNA OF SAND FLIES (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) IN CORUMBÁ, MATO GROSSO DO SUL, BRAZIL Jorge Espinoza Echeverria; Marinely Bustamante; Mery Diaz; Rudy Parrado; Ana Lineth Garciá Aline Etelvina Casaril1; Everton Falcão de Oliveira2; Neiva Zandonaide Monaco3; Gabriel Utida Eguchi4; Helen Rezende de Figueiredo1; Nathália Lopes Fontoura Mateus4; Wagner de Souza Fernandes1; Suellem Petilim Gomes1; Daiana Alovisi Souza5; Jucelei de Oliveira Moura Infran5,6; Elisa Teruya Oshiro6; Eunice Aparecida Bianchi Galati2; Alessandra Gutierrez de Oliveira1,5,6 IIBISMED, Facultad de Medicina, Universidad Mayor de San Simón, Cochabamba, Bolivia [email protected] Keywords: Leishmaniasis, Sand flies, Nycterohemeral, Peridomestic. The dynamics of different forms of the disease leishmaniasis is correlated with fluctuation of the populations of hosts and vectors, such as changes in the environment and climatic factors such as temperature, humidity and rainfall. Usually generated by the transformations of the wild environment through deforestation and replacement of wild animals by synanthropic, which facilitates the adaptation of some vector species colonized the environment and promoting an expansion of leishmaniasis transmission area of the forest to the home environment. Sand flies were captured and identified to species, from an area of Leishmania (Viannia) braziliensis endemicity in the Isiboro-Secure National Park in the Department of Cochabamba, during the season dry and humid, in peridomestic environments. Were determined the diversity of species during the nycthemeral rhytm of sand flies. The statistical analyzes beyond the ecological indices will be analyzed with the help of R package and Sigmaplot. Were captured 236 individuals belonging to 7 species of the genus Lutzomyia. Lu. shawi was the most abundant species during the two seasons with 52.1% in the dry season and 63.7% in the humid season. Was observed higher nycthemeral rhytm among the 20 and 21 hours; the activity of sand flies, decrease as a function to which the temperature decreases and relative humidity increases. During the humid season, present correlation positive among temperature and abundance (r2=0,744), but correlation negative among humidity and abundance (r2= 0,692). The diversity of species Shannon–Weaver, in the tow season showed, was of the H=0,603043 and equability of Pielou was the J= 0,87000. 103 1 Curso de Pós-Graduação em Doenças Infecciosas e Parasitárias - Faculdade de Medicina/ Universidade Federal de Mato Grosso do Sul; 2Departamento de Epidemiologia, Escola de Saúde Pública, Universidade de São Paulo; 3Centro de Controle de Zoonoses, Secretaria de Saúde do Município de Corumbá; 4Faculdade de Medicina Veterinária e Zootecnia - Universidade Federal de Mato Grosso do Sul; 5 Curso de Pós-Graduação em Biologia Animal - Centro de Ciências Biológicas e da Saúde/Universidade Federal de Mato Grosso do Sul; 6Laboratório de ParasitologiaCentro de Ciências Biológicas e da Saúde/Universidade Federal de Mato Grosso do Sul Cidade Universitária, 79070-900, Campo Grande, MS, Brasil. [email protected] Keywords: sand flies, abundance, diversity The municipality of Corumbá is considered area of intensive transmission for Visceral Leishmaniasis (VL) and reported 200 cases of the disease during 1999- 2011 and seven confirmed cases of American Cutaneous Leishmaniasis (ACL) from 2007 to 2011. Fauna studies leading up to the association between the occurrence of these insects and the prevalence of the leishmaniasis in the areas with records of the diseases and promoting measures for the control and surveillance. The aim of this study was to identify the sand flies (Diptera: Psychodidae: Phlebotominae) fauna in the Municipality of Corumbá, Mato Grosso do Sul, Brazil. The Sand flies were collected weekly with light automatic traps at 06:00 pm to 07:00 am, in peri and intradomicile environments in five urban districts, from April 2012 to March 2013. The sand flies were sent to the Laboratory of Parasitology at the Federal University of Mato Grosso do Sul. They were clarified and identified following the classification proposed by Galati (2003). The 390 weekly captures yielded 7,370 specimens of phlebotomine, 6,169 males (83.70%) and 1,201 females (16.30%). The fauna was composed by 12 species including: Brumptomyia brumpti, Evandromyia aldafalcaoae, Evandromyia cortelezzii, Evandromyia corumbaensis, Evandromyia sallesi, Evandromyia walkeri, Lutzomyia cruzi, Lutzomyia forattinii, Micropygomyia peresi, Martinsmyia oliveirai, Psathyromyia bigeniculata and Sciopemyia sordellii. The most abundant species was Lu. cruzi representing 93.79% and the second was Lu. forattinii with 3.53%. Overall, the insects were more frequently captured in peridomestic (60.45%) than indoors (39.55%) of the residences. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 89-P Ecology & Epidemiology ID 90-P Ecology & Epidemiology ENTOMOLOGICAL SURVEILLANCE OF PHLEBOTOMINE SAND FLIES VECTORS OF LEISHMANIASIS IN AREAS IMPACTED BY THE TRANSFER OF THE SÃO FRANCISCO RIVER PROJECT, IN THE STATE OF CEARÁ, BRAZIL THE INFLUENCE OF MOON LIGHT AND LUNAR PERIODICITY ON THE DENSITY OF PHLEBOTOMUS (LARROUSSIUS) ORIENTALIS PARROT, 1936 (DIPTERA: PSYCHODIDAE) FROM LIGHT TRAP COLLECTIONS IN ETHIOPIA Júlia dos Santos Silva1, Antonio Pereira dos Santos2, Fabrício Kássio Moura Santos2, Luiz Osvaldo Rodrigues da Silva2, Lindemberg Caranha3, Elizabeth Ferreira Rangel1 Araya Gebresilassie1, Oscar David Kirstein2, Aviad Moncaz2, Habte Tekie1, Meshesha Balkew3, Alon Warburg2, Asrat Hailu4, and Teshome Gebre-Michael3 1 1 Department of Zoological Sciences, Addis Ababa University, Addis Ababa, Ethiopia, Ethiopia; 2Department of Microbiology and Molecular Genetics, The Institute of Medical Research Israel-Canada The Kuvin Center for the Study of Infectious and Tropical Diseases, Faculty of Medicine, The Hebrew University, Hadassah Medical School, Jerusalem, Israel; 3Aklilu Lemma Institute of Pathobiology, Addis Ababa University, Addis Ababa, Ethiopia; 4Department of Microbiology, Immunology and Parasitology, Faculty of Medicine, Addis Ababa University, Addis Ababa, Ethiopia Laboratório de Transmissores de Leishmanioses, Instituto Oswaldo Cruz, FIOCRUZ, Av. Brasil, 4365, 21040-360, Manguinhos, Rio de Janeiro, Brazil; 2Secretaria de Saúde do Estado do Ceará, Brazil; 3Laboratório de Vetores, Reservatórios e Animais Peçonhentos Dr. Thomaz Correa Aragão, SESA/CE, Brazil. [email protected] Keywords: Phlebotominae, species diversity, environmental impact, vector ecology 104 [email protected] Keywords: Density, light traps, lunar cycles, P. orientalis American Cutaneous Leishmaniasis (ACL) and American Visceral Leishmaniasis (AVL) are relevant public health problems in Brazil. In addition to biological and physio-geographical factors, the environmental impact resulting from human actions, such as deforestation, construction of hydro-electric plants, migration processes, among others, play important roles in transmission of leishmaniasis. In Brazilian north-eastern state of Ceará, both ACL and AVL are endemic diseases, the latter presenting intense transmission. The Transfer of the São Francisco River project (TSFR) aims to transform the socio-economic context of the Northeast. However, some negative impacts have been discussed. From the point of view of public health, there is a hypothesis of epidemic outbreaks of ACL and AVL. The purpose of this study was the entomologic surveillance of Phlebotominae, to subsidise prevention and control actions for leishmaniasis in towns influenced by the TSFR in state of Ceará. Monitoring sites were selected in the towns of Jaguaretama, Jaguaribe, Lavras da Mangabeira, Brejo Santo, Mauriti, Iguatu and MissãoVelha. Sand flies were captured monthly from May 2011 to December 2013, with three HP light traps in each site, installed in and around household and in the woods, working for 12 hours from sunset for three consecutive nights. Data analyses used were indexes of richness, dominance, abundance and diversity of species. Taking into consideration the overall captures and analysing intra- and peridomiciliary areas and woods separately, Lutzomyia longipalpis (Lutz & Neiva, 1912), the main vector of AVL, was the dominant and most abundant species in all months, in all towns. In order, the other most frequent species were Lutzomyia lenti (Mangabeira, 1938), Lutzomyia evandroi (Costa Lima &Antunes, 1936) and Lutzomyia villelai (Mangabeira, 1942). Iguatu, a town with intense transmission of AVL, presented the highest number of specimens, of which 96.2% were Lu. longipalpis. The largest species richness was at Mauriti (S=13), while the greatest diversity was found in MissãoVelha (H´=1.26), towns with moderate transmission of AVL and ACL cases, registering Lutzomyia intermedia (Lutz & Neiva, 1912), Lutzomyia migonei (França, 1920) and Lutzomyia whitmani (Antunes & Coutinho, 1939), important vectors of ACL. Surveillance and control activities for AVL and ACL and education actions in the involved towns are advised to reduce the transmission risk. Financial support: FIOCRUZ Various environmental variables including moon light and lunar periodicity have been shown to affect the night activity of phototrophic dipteran flies. To investigate the possible effect of lunar phases on the activity of phlebotomine sand flies, trapping of sand fly species was conducted during December 2012 to June 2013 in Tahtay Adiyabo District, Northern Ethiopia. Sand flies were collected from peri-domestic and agricultural fields using CDC miniature light traps and sticky traps on 28 nights, divided among each of the 4 lunar phases. A total of 13,460 sand flies (11,633 in light traps and 1,827 in sticky traps) belonging to eight species of the genus Phlebotomus (P. orientalis, P. bergeroti, P. rodhaini, P. duboscqi, P. papatasi, P. martini, P. lesleyae and P. heischi) were captured. The predominant species in both trapping sites and in all lunar phases was P. orientalis, constituting 98.3% of the specimens. A statistically significant difference was observed in the density of P. orientalis collected by CDC light traps among lunar phases (P<0.05). The highest mean number (231.13±36.27 per trap/night) of P. orientalis was captured during the new moon phases, in the absence of moonlight. Significantly, fewer sand flies were attracted in light traps during full moon. However, P. orientalis and the other sand fly species from sticky traps did not differ in their density among the four lunar phases (P=0.122). The blood feeding of P. orientalis was not influenced by the presence or absence of moon light. Results of the current study showed that the attraction and trapping efficiency of CDC miniature light traps is substantially influenced by moon illumination. Accordingly, sampling of sand fly species using light traps to estimate their population density in the field should take the effect of moon light and lunar periodicity into account on the trapping efficacy of light traps. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 91-P Ecology & Epidemiology ID 92-P Vector Competence & Infections CHARACTERISTICS OF HUMAN DWELLINGS AND THEIR SURROUNDINGS THAT FAVOUR THE ABUNDANCE OF PHLEBOTOMINES IN FOUR URBAN AREAS OF THE CARIBBEAN COAST OF COLOMBIA PHLEBOTOMINE SAND FLY FAUNA (DIPTERA: PSYCHODIDAE) AND NATURAL FLAGELLATE INFECTIONS FROM AN AREA HIGHLY ENDEMIC FOR CUTANEOUS LEISHMANIASIS ON THE BRAZILIAN-BOLIVIAN FRONTIER IN THE STATE OF ACRE, BRAZIL Jorge Rodríguez-Jiménez1, Alveiro Pérez-Doria1, Luis Romero-Ricardo1, José Vergara1, Ana Montesino1, Lily Martínez1, Luis Cortés2, Suljey Cochero3 & Eduar E. Bejarano1 1 105 Iorlando da Rocha Barata, Maria Sueli Barros Pinheiro, Fábio Márcio Medeiros da Silva, Roberto Carlos Feitosa Brandão, Thiago Vasconcelos dos Santos, Fernando Tobias Silveira Grupo de Investigaciones Biomédicas, Universidad de Sucre, Sincelejo, Colombia. 2 Secretaría de Salud Departamental de Bolívar, Cartagena, Colombia. 3Secretaría de Salud Departamental de Sucre, Sincelejo, Colombia Instituto Evandro Chagas/ SVS/ MS, Ananindeua, Pa, Brazil [email protected] [email protected] Keywords: Lutzomyia, cutaneous leishmaniasis, urban transmission, Colombia Keywords: phlebotomine, flagellate, Amazonian frontier, cutaneous leishmaniasis The transmission of Leishmania is no longer exclusive to sylvatic and rural habitats, with new epidemiological scenarios arising in urban areas as a consequence of natural environmental changes or those brought about by human activities and favouring the survival or adaptation of sand fly vectors of the parasite. In the present study the characteristics of human dwellings and the peridomiciliary environment were analyzed in relation to the abundance of phlebotomines of the genus Lutzomyia in four urban areas of the Caribbean coast of Colombia, from which cases of cutaneous and visceral leishmaniasis had recently been reported. This involved the selection of 80 dwellings in the municipalities of Sincelejo, Ovejas, El Carmen de Bolívar and Cartagena. Visits were made to houses in which the characteristics of each dwelling and its surroundings were recorded. In addition insects were collected using CDC light traps hung inside and in the peridomiciliary area of each house from 18:00 to 06:00 on two consecutive nights. In all 9042 specimens of the genus Lutzomyia were captured belonging to 11 species, of which Lutzomyia evansi represented 93.3% of the total and was by far the most abundant in the four municipalities. The remaining phlebotomine fauna was represented by Lu. atroclavata (0.4%), Lu. carpenteri (0.04), Lu. cayennensis cayennensis (1.5%), Lu. dubitans (1.4%), Lu. gomezi (2.3%), Lu. panamensis (2.8%), Lu. rangeliana (0.2%), Lu. trinidadensis (0.4%), Lu. venezuelensis (0.4%) and Lu. micropyga (0.04%). Multiple correspondence analysis was used to determine that the following characteristics favoured the high abundance of phlebotomines in and around houses in these urban centres: (1) proximity (0-10 m) of dwellings to forested areas; (2) presence of bushes, grass and leaf litter in patios; (3) walls made of “bahareque” (adobe) and dirt floors; (4) accumulation of solid residues in open areas; (5) disposal of human waste in the open air; and (6) having electrical power in the dwellings. As measures to reduce phlebotomine density and the risk of urban Leishmania transmission, sanitary conditions in the study areas should be improved to eliminate sources of organic material in which immature sand flies develop. The construction of dwellings should also be modified to include other materials. Knowledge of the phlebotomine sand fly fauna in foci of cutaneous leishmaniasis (CL) is essential for epidemiological surveillance, providing information on the main existing species and their distribution. The increasing number of cases of CL in Amazonia, due mainly to environmental changes and high migratory flows, leads to the prioritization of surveillance at strategic areas, such as the Amazonian frontiers. Thus, the aim of this study was to identify the phlebotomine sand fly fauna and find natural flagellate infections in an area highly endemic for CL at the Brazilian-Bolivian frontier. The study was carried out in a rural area, namely Brasiléia municipality (S11°00'36” W68º44'52”), state of Acre (AC), Amazonian Brazil. During 2012 and 2013, four collections (15 days each) were undertaken in the dry and rainy seasons with eight CDC light traps disposed at ground level (1.5 meters) and two in the forest canopy (20m) from 18:00 to06:00 hours. Females were dissected under sterile conditions and each homogenate of infected midgut was twofold inoculated into Difco B45 media. Both sexes were identified according to taxonomic criteria (Young and Duncan, 1994). A total of 3,625 phlebotomine sand flies belonging to 41 species were collected. Females (n= 2,793) predominated over males (n= 1,026). The collections were more abundant in the rainy season (n= 2,716) than in the dry season (n = 1,103). Ten species were noted as being potentially implicated in the transmission of CL. Among them, the five most frequent were Lutzomyia whitmani (n = 878; 24.2%), Lu. auraensis (n= 767/ 21,1%), Lu. ubiquitalis (n= 289/ 7,9%), Lu. davisi (213/ 5,8%) and Lu. ntunesi (163;/ 4,4%). A single female of Lu. whithmani and two of Lu. yuilli yuilli were found harboring flagellates with a overall infection rate of 0,1%. No parasite isolation was accomplished successfully. In conclusion, this study leads to confirm the potential implication of Lu. whitmani and Lu. yuilli yuilli in the epidemiology of CL at AC and in addition, despite no natural infection was found in Lu. auraensis, its high frequency also adds weight to the hypothesis on the participation of this specie in the transmission of the disease, as has been suggested for Amazonian Peru. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 93-P Vector Competence & Infections ID 94-P Vector competence & Infections SANDFLIES (DIPTERA: PSYCHODIDAE) NATURALLY INFECTED BY LEISHMANIA IN RURAL AREAS OF THE CITY OF NOVA MUTUM, MATO GROSSO, BRASIL SAND FLY FAUNA (DIPTERA, PSYCHODIDAE) COLLECTED IN THE MIDDLE PURUS RIVER REGION, AMAZONAS, BRAZIL Sirlei Franck Thies1,2,3, Ana Lucia Maria Ribeiro3,5, Jorge Senatore Rodrigues Vargas3,5, Rosina Djunko Miyazaki4, Érika Monteiro Michalsky6, Edelberto Santos Dias6 1 Laboratory of Histology, Department of Basic Sciences in Health, Medicine College, UFMT, Cuiabá/MT. 2Regional Office of Health of Sinop, State Department of Health of Mato Grosso, Sinop/MT. 3Laboratory of Medical Entomology, Medicine College, UFMT, Cuiabá/MT. 4Laboratory of Leishmaniasis, Center of Research René Rachou/Fiocruz-Minas Gerais. 5University Hospital Júlio Müller, UFMT, Cuiabá/MT. 106 Túllio Romão R. da Silva, Mauro Diego Gobira Guimarães de Assis, Felipe Dutra Rêgo, Maíra Posteraro Freire, Paloma Helena Fernandes Shimabukuro Centro de Pesquisas René Rachou/FIOCRUZ MINAS [email protected] Keywords: Phlebotominae, Fauna, Amazonas State [email protected] Keywords: Leishmania, Lutzomyia, Sandflies. The leishmaniases are zoonotic diseases caused by protozoa of the Kinetoplastida order, Trypanosomatidae family, Leishmania genus that affect humans, and are transmitted in the Americas by insects belonging to the Lutzomyia genus, called sandflies. The present study aimed to identify the sandfly fauna and to verify their natural infection by Leishmania species in a rural area in the city of Nova Mutum, Mato Grosso, a place with notification of human cases of American Cutaneous Leishmaniasis (ACL). Entomological collections were made every two months, from June 2011 to April 2012, with CDC light traps which were installed at 6 p.m. and collected at 7 a.m. the next day, in ten specific places 100 meters distant from each other, from the edge to the inside of the forest. The captured insects were sorted, dissected, prepared and identified according to Young and Duncan (1994). Sandfly pools contained between 1 and 10 female insects, separated by species, trap number, place and date of capture. DNA extraction was performed with Extraction Kit of Tissues and Cells from GE Amersham Biosciences. Positive PCR bands of Leishmania species were purified using the commercial kit Qiagen. Genetic sequencing was performed via ABI 3130 platform. The Standard Nucleotide Blast program was used for analysis of bioinformatics and the sequences were compared to GenBank. The presence of 3,743 specimens belonging to 30 different species was found. The most abundant was Lutzomyia antunesi (45.44%), which was found in all months and capture places. Females constituted 73.15% of the specimens. The highest densities occurred in October 2011, during the rainy season in the state. Two hundred and ninetythree (293) pools of females were assembled, belonging to six different species, and L. antunesi was predominant (196/293). Thirteen (13) pools showed natural infection, corresponding to a minimum rate of infection of 1.01%. Lutzomyia ubiquitalis presented two pools infected either with Leishmania braziliensis or Leishmania guyanensis. Lutzomyia antunesi presented six pools infected either with Le. braziliensis or Le . guyanensis, four pools did not allow the alignment of the sequences (Leishmania species) and one pool infected with Leishmania infantum. These infected sandfly species are suspected vectors in the transmission cycle of ACL in this location, where the presence of Le. infantum poses the risk of transmission of Visceral Leishmaniasis. Phlebotomine sand flies are of medical importance because they are vectors of human pathogens, such as protozoa of the genus Leishmania Ross, etiological agent of American cutaneous leishmaniasis (ACL). In Lábrea, a municipality in the state of Amazonas, Brazil, ACL is primarily associated with subsistence activities, such as collection and extraction of forest products, undertaken by both indigenous and non-indigenous people. Data on ACL in indigenous populations are scarce, such that there is little information on the identity of the etiologic agent(s), reservoir host(s) and insect vector(s). The aim of this work was to study the sand fly fauna collected in the Indigenous Reserve Caititu, Lábrea. A total of 1,267 sand flies were collected, of which 819 (64.6 %) were females and 448 (35.4 %) males, from 10 genera and 32 species. The most abundant genera were Psychodopygus (34.3%), Trichophoromyia (22.9%)and Nyssomyia (15.3%). The most abundant species were Trichophoromyia ubiquitalis (Mangabeira) (n = 235, 18.5%), Psychodopygus davisi (Root) (n = 228, 18.0%) and Nyssomyia antunesi (Coutinho) (n = 135, 10.7%). Direct sequencing of PCR products demonstrated the presence of Leishmania (Leishmania) amazonensis and Leishmania (Viannia) braziliensis in the following species of sand flies: Evandromyia apurinan Shimabukuro, Silveira & Silva, Ny. umbratilis (Ward & Fraiha), Ny. yuilli yuilli (Young & Porter), Ps. davisi, Sciopemyia servulolimai (Damasceno & Causey) and Th. ubiquitalis. The presence of natural infection by Leishmania detected in the sand fly species investigated in this study suggests their possible role in the transmission cycle of ACL in the study area. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 95-P Vector competence & Infections ID 96-P Vector competence & Infections MOLECULAR DETECTION OF LEISHMANIA IN SAND FLIES (DIPTERA: PSYCHODIDAE) AND S PAT I A L D I S T R I B U T I O N O F A M E R I C A N CUTANEOUS LEISHMANIASIS IN THE XAKRIABÁ INDIGENOUS RESERVE, BRAZIL FIRST REPORT OF NATURAL PROMASTIGOTE INFECTION OF SERGENTOMYIA DREYFUSSI (DIPTERA: PSYCHODIDAE) SAND FLIES IN AN ENDEMIC FOCUS OF CUTANEOUS LEISHMANIASIS IN CENTRAL TUNISIA Felipe Dutra Rego, Jerônimo Marteleto Nunes Rugani, Patrícia Flávia Quaresma, Paloma Helena Fernandes Shimabukuro, Gabriel Barbosa Tonelli, Ronaldo Guilherme Carvalho Scholte, Célia Maria Ferreira Gontijo Jomaa Chemkhi1, Rihab Yazidi2, Mahdi Driss3, Jihene Bettaieb2 and Afif Ben Salah2 Grupo de Estudos em Leishmanioses. Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz. Belo Horizonte, Minas Gerais, Brasil. 107 1 Laboratoire d'Epidémiologie Moléculaire et Pathologie Expérimentale Appliquée Aux Maladies Infectieuses, Institut Pasteur de Tunis, Université Tunis El Manar, Tunisia. 2 Laboratoire d'Epidémiologie Médicale, Institut Pasteur de Tunis, Université Tunis El Manar, Tunisia. 3 Plateforme de séquençage, Institut Pasteur de Tunis, Université Tunis El Manar, Tunisia. [email protected] [email protected] Keywords: Phlebotominae, Leishmania infection, Indigenous Reserve, Kernel Density Estimator; Autochthonous cases of American Cutaneous Leishmaniasis (ACL) are reported since 2001 in Xakriabá Indigenous Reserve located at São João das Missões, north of Minas Gerais state, characterizing this area as endemic for this disease. This study aimed to investigate the association between sand flies and the transmission of Leishmania species. The data were correlated with epidemiological aspects previously reported (natural infection in vertebrate hosts and human cases of ACL) to identify hotspots related to infection by Leishmania braziliensis, the most prevalent Leishmania species in the study area. The sand flies were captured with HP light traps bimonthly for three full days between October 2011 and August 2012 on trails previously selected for the study of Leishmania reservoirs and sand fly fauna. The DNA was extracted in pools, containing up to ten females from same species and site colletion. The PCR was targeted to the internal transcribed spacer 1 (ITS1) and amplicons were digested by the HaeIII enzyme to analyze their restriction-fragment length polymorphisms (RFLP). The spatial distribution of the ACL was analysed by Kernel density to define the hotspots - concentration and pattern of ACL infection - in the study area. A total of 4,031 female sand flies were captured and grouped in 1,026 pools. Twelve pools were PCRpositive (1.16%) and Leishmania braziliensis infections were found in Nyssomyia intermedia, Martinsmyia minasensis, Micropygomyia capixaba and Micropygomyia peresi. Other Leishmania species (Leishmania guyanensis, Leishmania infantum and Leishmania amazonensis) were found in Evandromyia lenti, Mt. minasensis, Mi. peresi and Ny. intermedia. The spatial analyses identify a main hotspot indicating the location where the presence of Leishmania braziliensis was more frequent. The results of natural infection in different hosts studied by spatial analysis is an important tool to assess the areas with high prevalence of parasite infection and may contribute to the choice of specific preventive and control measures. Keywords: Flagellate, infection, Sergentomyia, Tunisia In order to study the natural infection of phlebotomine sandflies by Leishmania parasites in central Tunisia where cutaneous leishmaniasis is endemic, an entomological survey was carried out during the transmission period from July to November 2013, in the region of Mnara, belonging to the governorate of Kairouan. A total of 234 Phlebotomus papatasi, 89 P. longicuspis, 54 P. perniciosus, 44 Sergentomyia (antennata, 34 S. fallax, 28 S. dreyfussi and 12 S. munita parroti were captured from Psammomys obesus burrows, houses and animal shelters in a chenopod biotope, using CDC light traps. Female sand flies were first dissected to look for natural parasite infection in all parts of the gut. Then, they were morphologically identified based on the keys of Lewis and Léger (1983). Out of 230 females sand flies dissected, one identified as S. dreyfussi and caught in an old house harboring various reptile lizards, was found containing flagellates within the digestive tract. In vitro isolation and culture of these promastigote forms allowed further DNA extraction to be used for molecular analysis. PCR amplification followed by restriction fragment length polymorphism (RFLP) targeting ribosomal Internal Transcribed Spacer 1 (ITS1) DNA and sequencing analyses indicated that the observed promastigotes are probably reptile parasites belonging to the Trypanosomatidae family. This study constitutes the first report in Tunisia on the natural promastigote infection of Sergentomyia sand flies. This raises questions about the precise promastigote species identification, and more specifically to the possibility of the presence of Sauroleishmania, suggested by the proximity to lizard habitats. Further molecular analyses are ongoing to confirm this hypothesis. This study received support from In Bio International Inc. grant N°: W81XWH-09-C-0079. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 97-P Vector competence & Infections ID 98-P Vector competence & Infections SALIVARY GLAND HOMOGENATES FROM WILDCAUGHT LUTZOMYIA FLAVISCUTELLATA AND L. (PSYCHODOPYGUS) COMPLEXUS DID NOT EXACERBATE LEISHMANIA (L.) AMAZONENSIS AND L. (V.) BRAZILIENSIS INFECTION IN BALB/C MICE NATURAL INFECTION OF PHLEBOTOMINE SAND FLIES (DIPTERA: PSYCHODIDAE) BY LEISHMANIA ROSS (KINETOPLASTIDA: TRYPANOSOMATIDAE) IN “TERRA FIRME” AND “VÁRZEA” ENVIROMENTS IN TEFÉ MUNICIPALITY, AMAZONAS STATE, BRAZIL Laurenti MD, Francesquini FC, Passero LF, Tomokane TY, Carvalho AK, Corbett CEP, Silveira FT Medical School, University of São Paulo [email protected] Keywords: Experimental Cutaneous Leishmaniasis, Pathology, Lutzomyia flaviscutellata, Lutzomyia (Psychodopygus) complexus 108 Antonio Marque Pereira Júnior1,2; Carolina Bioni Garcia Teles3, Ana Paula de Azevedo dos Santos3, Felipe Arley Costa Pessoa2, Jansen Fernandes de Medeiros1,3 1 Inststuto nacional de Pesquisas da Amazônia, 2Centro de Pesquisa Leônidas & Maria Deane, FIOCRUZ AMAZÔNIA, 3FIOCRUZ Noroeste. [email protected] Keywords: Epidemiology, vectors, Leishmania lainsoni During the natural transmission of leishmaniasis, the infected female phlebotomine regurgitates promastigotes into the host's skin together with the saliva. It has been reported that components of vector saliva contain immunomodulatory properties that facilitate the establishment of infection in the host, however most studies employed salivary gland lysate (SGL) of laboratory colonized vectors. Thus, the main objective of this study was to evaluate the specificity of SGL of the phlebotomines Lutzomyia flaviscutellata and Lutzomyia (Psychodopygus) complexus caught in the field for the infectivity of L. (L.) amazonensis and L. (V.) braziliensis. BALB/c mice were inoculated in the hind footpad with promastigotes of L. (L.) amazonensis and L. (V.) braziliensis in the absence or presence of L. flaviscutelata, and L. (P.) complexus SGL. The evolution of the lesion size was evaluated weekly and biopsies from the site of infection were collected for histopathological analysis and determination of parasite load in the 4th and 8th week post infection (PI). Draining lymph nodes were also collected to characterize subsets of T cells by flow cytometry. L. (L.) amazonensis and L. (V.) braziliensis infections were not exacerbated in the groups of mice co-inoculated with the SGL of either phlebotomine species, in relation to the control group inoculated only with the parasite. The lesion size and parasite burden of the control group was higher or equal to the groups with saliva. In L. (L.) amazonensis infections there was a decrease of CD4+ cells and an increase in the population of CD8+ cells, while in L. (V.) braziliensis infections there was a maintenance of CD4+ cells and an increase in the population of CD8+ cells in all groups compared with the control group. As a whole, the results show that the saliva of L. flaviscutellata and P. complexus, in the natural vector/parasite binomium or not, did not favor the establishment of the infection caused by L. (L.) amazonensis e L. (V.) braziliensis in BALB/c mice. Supported by CNPq, FAPESP and LIM50 HC-FMUSP. Phlebotomine sandflies are insects of medical importance because some species play a role in transmission of leishmaniasis. The aim of this work was to detect natural infection and possible vectors in Tefé municipality, Amazonas state, Brazil. Phlebotomine used were collected using HP light traps during six months: January, February, April, August, September and October, of 2013. To verify natural infection, DNA samples were submitted to Polymerase chain reaction (PCR) using kDNA gene and the identification of Leishmania species were performed by PCR RFLP with hsp70 gene and genetic sequence. A total of 1.679 was grouped in 60 samples of Trichophoromyia ubiquitalis, 18 pools of Nyssomyia antunesi, one pool of Ny. umbratilis, eight of Ny. yuilli yuilli, four of Psychodopygus ayrozai and four of Ps. davisi. In kDNA region 14/95 samples were positive to presence Leishmania spp., ten sample belonging to Th. ubiquitalis and four sample to Ps. davisi. The minimal infection rate (MR) was 0.83%. In PCR to hsp70 gene was observed seven sample positive of the 14 sample amplified for kDNA. This sample belonging to Th. ubiquitalis. In PCR RFLP was observed that samples submitted to Hae III enzyme show a pattern between Leishmania (Viannia) lainsoni and Leishmania (V.) shawi. The genetic analysis confirmed the specie L. (V.) lainsoni in Th. ubiquitalis sample. The similarity of sequences was 99% compared to reference strain deposited on GenBank and less of 1% divergence. The sequence analysis makes this work the first report of Th. ubiquitalis infected by L. (V.) lainsoni in Amazonas state. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 109 ID 99-P Vector competence & Infections ID 100-P Vector competence & Infections TRANSMISSION OF LEISHMANIA INFANTUM TO HAMSTERS THROUGH THE BITE OF EXPERIMENTALLY INFECTED PHLEBOTOMUS PERNICIOSUS AND MONITORING OF ANIMALS INFECTIVITY BY XENODIAGNOSIS PHYLOGENETIC ANALYSIS OF THE LEISHMANIA ENRIETTII COMPLEX AND SPECULATIONS ON THEIR VECTOR BIOLOGY P.A. Bates, M.D. Bates, G. Kwakye-Nuako Inés Martín-Martín, Maribel Jiménez, Estela González, Ricardo Molina Lancaster University, UK [email protected] Unidad de Entomología Médica, Servicio de Parasitología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Ctra. Majadahonda-Pozuelo s/n, 28220, Majadahonda, Madrid, Spain. Keywords: Leishmania enriettii, phylogenetics, sand flies, midges [email protected] Keywords: Phlebotomus perniciosus, Leishmania infantum, natural transmission, xenodiagnosis A leishmaniasis outbreak is currently occurring in south western Madrid region (Spain) and already accounts for more than 560 human cases. The causative agent is Leishmania infantum and the main vector in the area is Phlebotomus perniciosus. All Leishmania isolates from the focus have the same genotype as the strain MHOM/ES/87/LOMBARDI but no association of isolate types and virulence has been demonstrated. Our aim was to perform natural transmission studies in animal models using isolates from P. perniciosus collected in the field (IPER/ES/2012/BOS1FL1 and IPER/ES/2012/POL2FL6) during the entomological survey carried out in the transmission season of 2012 in the human leishmaniasis outbreak area simultaneously with a well characterized L. infantum strain (MCAN/ES/98/LLM-877).Two colonies of P. perniciosus established from specimens captured in Madrid were experimentally infected with 2.5x107 promastigotes/ml of L. infantum isolates. Ten days after experimental infections, sand flies were fed on hamsters. In addition, other hamsters were inoculated with 107 promastigotes/ml of the same cultures. Effective transmissions by bite were monitored and compared with intraperitoneal infections. Experimental infections were successful but natural transmission by sand fly bite was only achieved with isolates from the outbreak. Even attempts of biting - without blood ingestion by infected sand flies with the focus isolate - led to a transmission of the parasite. Infected hamsters were xenodiagnosed every 2 months and were infective to sand flies as soon as 2 months post-transmission experiments. Hamsters inoculated with parasites from the outbreak presented clinical visceral disease and were also infective to sand flies when they were tested by xenodiagnosis. Vector transmission generated a slower progression of clinical disease than intraperitoneal infection, but both were infective to P. perniciosus by xenodiagnosis studies. A visceral leishmaniasis model that mimic the natural conditions of transmission in nature was set and allowed us to compare Leishmania strain virulence and infectivity to sand flies. The differences found in the outcome of the disease and infectivity to sand flies of infected hamsters by strains that are circulating in the focus would contribute to the understanding of their role in the epidemiology of the human leishmaniasis outbreak in Madrid.This study supported by the Spanish Project AGL2008-01592 and by EU grant FP7-2011-261504 EDENext. Recent work has provided evidence for a new phylogenetic grouping of Leishmania parasites, the Leishmania. enriettii complex, that may represent a new Leishmania subgenus in addition to Leishmania, Viannia and Sauroleishmania. Known members of the L. enriettii complex include L. enriettii itself (from Brazil), L. martiniquensis (from Martinique, a Caribbean island) and L. australiensis (from Australia). To these we now add further species from Namibia, Ghana and Thailand. Some of these parasites are human pathogens, whilst others appear to be confined to wild animals. The very wide geographical spread of the L. enriettii complex, more widely distributed than any of the existing subgenera, together with their branching from the base of the Leishmania phylogenetic tree, both suggest an early branching and ancient lineage. The vectors of the L. enriettii complex have not been established with certainty for any member. It has been speculated that L. enriettii has a phlebotomine sand fly vector, however, this has not been proven, and L. enriettii appears not able to establish infection in Lutzomyia longipalpis, a widely used permissive sand fly host. For the other members there is little evidence available, except for L. australiensis. In this species evidence to date supports the vectors to be day-biting midges of the genus Forcipmyia, although transmission by midge-bite has not been attempted yet. It therefore appears that some, or maybe even all, of the L. enriettii complex may have non-sand fly vectors. This speculation remains to be proven or disproven, with efforts currently underway in both Ghana and Thailand to identify potential vectors. However, if true, this would require a re-definition of the genus Leishmania to include non-sand fly transmission or the creation of a new genus to accomodate the non-sand fly transmitted parasites. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 101-P Vector competence & Infections ID 102-P Vector competence & Infections VECTOR COMPETENCE OF NYSSOMYIA WHITMANI (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) TO LEISHMANIA (LEISHMANIA) AMAZONENSIS N AT U R A L I N F E C T I O N O F L U T Z O M Y I A (NYSSOMYIA) Y. YUILLI AND L. ( T R I C H O P H O R O M Y I A ) O C TAV I O I W I T H LEISHMANIA SPP. IN AGUAS CALIENTES, LA CONVENCION, CUSCO, PERÚ. Magda Freitas Fernandes1, Kleiton Maciel dos Santos1, Antonio Carlos Ferrari Júnior1, Ana Paula Silva Levay1, Fábio Juliano Negrão1, Wedson Desidério Fernandes1, Elisa Teruya Oshiro2, Maria Elizabeth Moraes Cavalheiros Dorval2, José Dilermando Andrade Filho3, Alessandra Gutierrez de Oliveira2, Eunice Aparecida Bianchi Galati4 1 Universidade Federal da Grande Dourados (UFGD), Mato Grosso do Sul, Brazil. 2 Universidade Federal de Mato Grosso do Sul (UFMS), Brazil. 3Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz (Fiocruz), Minas Gerais, Brazil. 4Universidade de São Paulo (USP), São Paulo, Brazil. [email protected] 110 D. Rado1, D. Pita-Pereira4, T. Araujo Pereira4, MM Quispe-Florez1, L. Brahim3, E. Rangel3, B. Valladares2, R. Pacheco1 1 Instituto Universitario de Enfermedades Tropicales y Biomedicina del Cusco – UNSAAC, Perú; 2Instituto Universitario de Enfermedades Tropicales y Salud Publica de Canarias, Universidad de La Laguna, Tenerife, España; 3Laboratório de Transmissores de Leishmanioses – Instituto Oswaldo Cruz/FIOCRUZ, Brasil; 4 Laboratório de Biologia Molecular e Doenças Endêmicas – Instituto Oswaldo Cruz/FIOCRUZ, Brasil [email protected] Keywords: Vector, Competence, Nyssomyia whitmani, Leishmania amazonenses Findings of phlebotomine species naturally infected by Leishmania spp., and not yet implicated in the transmission of particular leishmaniasis agents, bring out the need to investigate those species' vectorial competence. The objective of this study was to investigate the experimental susceptibility of Nyssomyia whitmani to Leishmania (Leishmania) amazonensis and its transmission to uninfected (susceptible) hamsters. Wild phlebotomine species were captured in a forest fragment in Dourados, MS, during the night period with black modified Shannon traps and the use of electrical aspirators. Males and females were kept in cages for mating and the blood meal was taken on susceptible hamsters exposed to biting for two hours. After 48 hours the engorged females were placed in individual polyethylene vials for egg laying. The eggs were then transferred to small Petri dishes until the emergence of the adult specimens. The susceptibility tests with Ny. whitmani F1 females were undertaken in two stages: in the first experiment 156 specimens emerged in an average period of 55 days. Of the 92 females, 63 (68.5%) took the blood meal on the infected hamsters. Of those 63, 22 (34.9%) took the second meal on the susceptible hamsters: 13 (20.6%) on the 5th day, 5 (7.9%) on the 6th day, 2 (3.2%) on the 12th day and 2 (3.2%) on the 21st day. In the second experiment 713 specimens emerged in an average period of 58 days. 561 females took the blood meal on the infected hamsters, and 131 (23.4%) the second blood meal on the susceptible hamsters: 13 (2.3%) on the 6th day, 82 (14.6%) on the 7th day, 6 (1.1%) on the 8th day, 19 (3.4%) on the 9th day, 6 (1.1%) on the 10th day, 3 (0.5%) on the 12th day and 2 (0.4%) on the 14th day. A total of 27 susceptible hamsters were exposed to the females which had fed themselves on the infected hamsters. The confirmation of infection by Leishmania in the hamsters exposed for the 2nd blood meal was undertaken by Polymerase Chain Reaction to kDNA followed by Restriction Fragment Length Polymorphism Assay with Hae III of the samples of the spleen of the 27 hamsters, giving a 100% positive reaction. Seeing that Ny. whitmani was experimentally infected by L. amazonensis and succeeded in transmitting the infection to susceptible hamsters, its vectorial competence for this parasite has thus been demonstrated . However, it is necessary to undertake studies for the assessment of its vectorial capacity in regard to the transmission of L. amazonensis. Keywords: Natural Infection, L. y. yuilli , L. octavioi, Leishmania spp. Aguas Calientes (at 890 m) is an important endemic area of leishmaniosis in the province of La Convencion (Cusco, Perú). The most frequently species found in the area are L. y. yuilli (47,25%) and L. octavioi (25,97%).With the aim of determine natural Leishmania spp. infection in this species, we analized, by PCR multiplex, 550 females of . L. y. yuilli and 170 females of L. octavioi (grouped in pools of 10 individuals) that come from collection made in intra, peri and extra domestic environments with Shannon and CDC traps during dry and rainy seasons in 2010-2011. Positive result were detected in seven pools of L. y. yuilli (from extradomiciliary collection) and three pools of L. octavioi (from intra, peri and extradomiciliary collection) with a minimum infection rate of 1,27% and 1,76%, respectively. In the studied area, both species could be involved in Cutaneus Leishmaniosis epidemiology.Support by IOCFIOCRUZ, Brazil; CNPq, Brazil and AECID, Spain. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 103-P Vector competence & Infections ID 104-P Vector competence & Infections LUTZOMYIA (NYSSOMYIA) Y. YUILLI AND L. (TRICHOPHOROMYIA) SP. FROM ALTO IVOCHOTE, LA CONVENCIÓN, CUSCO, PERU AGGREGATED PATTERN OF Leishmania infantum chagasi TRANSMISSION FROM NATURALLY INFECTED DOGS TO LABORATORY REARED Lutzomyia longipalpis THROUGH XENODIAGNOSIS Anita Quispe1, MA Quispe-Ricalde2, D. Rado1, MM Quispe-Florez1, M. Soto1, Pérez JE3, B. Valladares2, R. Pacheco1 1 Instituto Universitario de Enfermedades Tropicales y Biomedicina del Cusco – UNSAAC, Perú; 2Instituto Universitario de Enfermedades Tropicales y Salud Publica de Canarias, Universidad de La Laguna, Tenerife, España; 3Instituto de Medicina Tropical Alexander von Humboldt, UPCH, Lima, Perú [email protected] Keywords: Leishmaniosis, Lutzomyia, PCR RFLP hsp70, natural infection Alto Ivochote is an endemic area for leishmaniosis and Lutzomyia spp. in large populations are frequent in the area. The objective of this study was to combine entomological with molecular techniques to characterize Leishmania spp. detected in Lutzomyia (Nyssomyia) y. yuilli y Lutzomyia (Trichophoromyia) sp., as they are the most frequently species for the study area. From collection made during the rainy and dry seasons of 2011, 594 females of Lu. (N.) y. yuilli and 458 females of Lu. (T.) sp., were studied to determine the natural infection with Leishmania and species characterization through PCR – RFLP hsp70. Both species were found naturally infected by Leishmania (Viannia) sp. with natural infection rates of 18.35% and 3.57%, respectively. On the other hand Leishmania (V.) braziliensis was detected and characterized in 19 females of Lu. (N.) y. yuilli from extra domestic collection with Shannon trap. These results suggest that Lu. (N.) y. yuilli play an important role in the transmission cycle of L. (V.) braziliensis to man and animals in Alto Ivochote. 111 Andre Antonio Cutolo1, Fredy Galvis Ovallos2, Cecilia Oliveira Lavitschka2, Mariana Dantas da Silva2, Leticia Maria Molla3, Fabio dos Santos Nogueira3, Vera Lucia Pereira-Chioccola1, Ingrid Menz3, Eunice Aparecida Bianchi Galati2 1 Laboratorio de Biologia Molecular de Parasitas e Fungos, Instituto Adolfo Lutz, São Paulo, Brazil. 2 Departamento de Epidemiologia, Faculdade de Saúde Pública, USP, São Paulo, Brazil. 3 DVM Independent Researcher, São Paulo, Brazil [email protected] Keywords: Xenodiagnosis, Visceral Leishmaniasis, Vector Competency, Transmission Rate Thirty six Visceral Leishmaniasis symptomatic dogs, naturally infected with Leishmania infantum chagasi were evaluated through xenodiagnosis in order to check infectivity rate to laboratory reared Lutzomyia longipalpis nulliparous females. Dogs were from an American Visceral Leishmaniasis endemic area in São Paulo state, Brazil, and confirmatory diagnosis for Leishmania infection was performed through serology, cytology and molecular methods. All animals were considered polysymptomatic at the time of xenodiagnosis. Sandflies were dissected five days after bloodfeeding under light microscopy in order to detect live Leishmania epimastigotes and promastigotes in the gut contents. Sandfly infection was scored as low (+), medium (++) and high (+++) level depending on the number of protozoa observed. Among the 36 dogs evaluated, 19 (52.8%) infected sandflies. An average of 22 insects were dissected per exposed dog, with 805 sandflies dissected in total and 99 (12.3%) found infected. Positive dogs had different levels of infectivity to sandflies. Individual dog infectivity rates ranged from 4% (1/25) to 95% (19/20) of dissected sandflies. Fourteen (73.7%) positive dogs had infectivity rates ranging from 4% to 20%, as following: 4.0% (1/25), 4.8% (1/21), 5.0% (1/20), 5.0% (1/20), 5.0% (1/20), 6.1% (2/33), 6.7% (1/15), 10.0% (2/10), 12.5% (2/16), 12.5% (3/24), 14.3% (3/21), 15.0% (3/20), 19.1% (4/21) and 20.0% (4/20). In addition, these 14 dogs generally infected sandflies at a low intensity, with only one sandfly being scored as +++. Five (26.3%) dogs were the most infectious animals with 60.0% (6/10), 65.0% (13/20), 75.0% (15/20), 85.0% (17/20) and 95.0% (19/20) infected sandflies per exposure, and these dogs also generally infected sandflies at a higher intensity. Among the 99 infected sandflies, 29 (29.3%) came from the fourteen lowest infectious dogs and 70 (70.7%) sandflies originated from the five most infectious dogs. An aggregated pattern of Leishmania infantum chagasi transmission from dogs to sandflies was observed in this study. This finding suggests that identification of the most infectious dogs within a population and consequently use of proper transmission blocking tools directed to them, may be an essential feature of transmission control for this zoonotic visceral leishmaniasis causing agent in Brazil. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 105-P Vector competence & Infections ID 106-P Genome & Genetics V E C TO R S O F L E I S H M A N I A M E X I C A N A I N SOUTHEASTERN OF MEXICO GENETIC CHARACTERIZATION OF NORTHERN AND SOUTHERN POPULATIONS OF PHLEBOTOMUS SERGENTI (DIPTERA: PSYCHODIDAE) POPULATIONS IN MOROCCO Pech-May Angélica1, Sánchez-García Laura2, Berzunza-Cruz Miriam2, Becker-Fauser Ingeborg2, Escobedo-Ortegón Javier3, Rebollar-Téllez Eduardo A4 1 Centro Regional de Investigación en Salud Pública -INSP, Tapachula, Chiapas, México. 2Departamento de Medicina Experimental, Facultad de Medicina, Universidad Nacional Autónoma de México, Hospital General de México, México, D.F., México. 3Departamento de Enfermedades Infecciosas y Transmitidas por vector, Centro de Investigaciones Regionales Dr. Hideyo Noguchi, Universidad Autónoma de Yucatán, Mérida, Yucatán, México. 4Laboratorio de Entomología Médica, Departamento de Zoología de Invertebrados, Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León, Monterrey, Nuevo León, México. [email protected] In Mexico, leishmaniases represents a public health problem. Leishmania (Leishmania) mexicana has so far been identified as the main ethologic agent of cutaneous leishmaniasis (LCL) along the country. The southeaster of Mexico is the main endemic area for LCL and it has been demonstrated that Bichromomyia olmeca olmeca is the primary vector, however Lutzomyia cruciata, Psychodopygus panamensis and Psathyromyia shannoni have also been found infected by PCR with Le. (Le.) mexicana. Therefore we suggested that these species may be acting as primary and/or secondary vectors in some areas of Yucatan Peninsula (YP), Mexico. Here we present a resume of the state of the knowledge of the abundance, sandfly biting rates and infection rate of Le. (Le.) mexicana vectors in YP. The results of the abundance per species in each location indicated that the most abundant species in the YP are Bi.o. olmeca (66%), followed by Pa. shannoni (52%) and Ps. panamensis (43%). Regarding to the monthly biting rate, the results showed that it depends of each species and locality. The mean biting rate per hour, showed peak from 18:00 to 19:00 in all localities. The high infection rate was in 20JUN (5.3%), the sandfly Pa. shannoni was infected in all localities, while that only sandfly Bi. o. olmeca was infected in collections made in Shannon and Disney traps. We can propose according with the sand fly seasonal abundance distribution and the biting rates, those biotic and abiotic factors, which vary within a period time, affects sand fly populations. 112 Boussaa Samia1,2; Ouanaimi Fouad1; Kahime Koloud1; Echchakery Mohamed1; Boumezzough Ali1 1 Equipe d'Ecologie Animale-Environnement, Faculté des Sciences Semlalia, Université Cadi Ayyad, Marrakech, Maroc; 2ISPITS-Institut Supérieur des Professions Infirmières et des Techniques de Santé, Ministère de Santé, Marrakech, Morocco [email protected] Keywords: Phlebotomus sergenti, Isoenzyme, Mitochondrial DNA, Morocco In Morocco, Phlebotomus sergenti is the proven vector of Leishmania tropica, a causative agent of cutaneous leishmaniasis (Guilvard et al., 1991). Many Moroccan L. tropica foci have been reported, in the rural areas of Azilal, Essaouira and Chichaoua (Pratlong et al., 1991; Guernaoui et al., 2005; Ramaoui et al., 2008), but also in urban and peri-urban areas of Taza and Zouagha Moulay Yaacoub (Guessous-Idrissi, 1999; Rhajaoui et al., 2004). The taxon known as P. sergenti shows considerable genetic variation over its large global area of distribution (Depaquit et al., 2002; Yahia et al., 2004; Moin-Vaziri et al., 2007). This study presents a genetic comparison, using isoenzyme and molecular tools, between northern (Chefchaouene, Ouezzane, and Taounate) and southern (Marrakech, Ourika, Rhmate and Touama) populations of Moroccan P. sergenti. For morphological studies sandflies were collected on sticky-paper traps. After dissection, sandflies were slide-mounted in Marc Andre solution and examined by light microscopy. For isoenzyme and molecular analyses, specimens were collected in CDC light traps and immediately stored at -80°C. Ten isoenzyme systems were tested and the qualitative analysis of zymogram profiles revealed nine polymorphic loci (HK, PGM, PGI, 6PGD, FUM, MDH1, ME, ICD2 and ACO). A section of the mitochondrial gene Cytochrome b (Cyt b) was amplified by PCR with the primer pair CB3-FC/CB-R06. The calculation of allele frequencies, tests for deviation from Hardy–Weinberg equilibrium, and phenetic analysis were performed using Genepop and Biosys2 programs. Our results showed that these two P. sergenti populations do not have the genetic characteristics of a single species, as indicated by the HW disequilibria for several tested loci and by the presence of two P. sergenti mitochondrial lineages. The epidemiological significance of this genetic variability merits future investigation. References: Depaquit et al., 2002. Int J Parasitol, 32, 1123–1131. Guernaoui et al., 2005. J M Entomol, 42, 697–701. Guessous-Idrissi, 1999. Recherche Nationale, les Cahiers du Médecine, 2, 37–40.Guilvard et al., 1991. Ann Parasitol Hum Comp, 66, 96–99. Moin-Vaziri et al., 2007. Acta Trop, 102, 29–37.Pratlong et al., 1991. Ann Parasitol Hum Comp, 66, 100–104. Ramaoui et al., 2008. Parasitol Res, 103, 859–863. Rhajaoui et al., 2004. Trans R Soc Trop Med Hyg, 98, 299–301. Yahia et al., 2004. Parasite, 11, 189–199. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 113 ID 107-P Genome & Genetics ID 108-P Leishmania - Sand fly Interactions EVIDENCE FOR GENETIC DIFFERENTIATION AT THE MICROGEOGRAPHIC SCALE IN PHLEBOTOMUS PAPATASI POPULATIONS FROM SUDAN ANALYSIS OF CARBOHYDRATE EPITOPES ON THE S U R FA C E O F M I D G U T E P I T H E L I U M O F LUTZOMYIA (LUTZOMYIA) LONGIPALPIS AND LUTZOMYIA (NYSSOMYIA) ANTUNESI (DIPTERA, PSYCHODIDAE) FROM PARÁ, STATE AMAZONIAN BRAZIL Noteila M Khalid, Marium A Aboud, Fathi M Alrabba, Dia-Eldin A Elnaiem and Frederic Tripet Department of Zoology, Khartoum College of Medical sciences, Khartoum, Sudan. [email protected] Keywords: Phlebotomus papatasi, Sudan, Gene flow, Genetic differentiation. Background: Cutaneous Leishmaniasis (CL) is endemic in Sudan. It is caused by Leishmania major parasites and transmitted by Phlebotomus papatasi sandflies. Recently, uncommon clinical manifestations of CL have been reported. Moreover, L. donovani parasites that cause Visceral Leishmaniasis (VL) have been isolated from CL lesions of some patients who contracted the disease in Khartoum State, Central Sudan with no history of travelling to VL endemic sites on south-eastern Sudan. Because different clinical manifestations and the parasite behaviour could be related to genetic differentiation, or even sub-structuring within sandfly vector populations, a population genetic study was conducted on P. papatasi populations collected from different localities in Khartoum State known for their uncommon CL cases and characterized by contrasting environmental conditions. Methods: A set of seven microsatellite loci was used to investigate the population structure of P. papatasi samples collected from different localities in Khartoum State, Central Sudan. Populations from Kassala State, Eastern Sudan and Egypt were also included in the analyses as outgroups. The level of genetic diversity and genetic differentiation among natural populations of P. papatasi was determined using FST statistics and Bayesian assignments. Results: Genetic analyses revealed significant genetic differentiation (FST) between the Sudanese and the Egyptian populations. Within the Sudanese P. papatasi populations, one population from Gerif West, Khartoum State, exhibited significant genetic differentiation from all other populations including those collected as near as 22 km. Conclusion: The significant genetic differentiation of Gerif West P. papatasi population from other Sudanese populations may have important implication for the epidemiology of leishmaniasis in Khartoum State and needs to be further investigated. Primarily, it could be linked to the unique location of Gerif West which is confined by the River Nile and its tributaries that may act as a natural barrier for gene flow between this site and the other rural sites. The observed high migration rates and lack of genetic differentiation among the other P. papatasi populations could be attributed to the continuous human and cattle movement between these localities. Oliveira, D.M.S.1; Lima, J.A.N2.; Silva, B.J.M 1; Farias, L.H.S1; Santos, T.V2.; Silveira, F.T2; Silva, E.O1 ¹Laboratório de Biologia Estrutural/Laboratório de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal do Pará, Brazil.2 Instituto Evandro Chagas - Ministério da Saúde [email protected] Key-Words: Helix pomatia lectin, Lutzomyia, glycoprotein. Leishmaniasis Leishmaniases are worldwide diseases that occur in 98 countries. In Brazil, cases of American Visceral Leishmaniasis (AVL) and Cutaneous Leishmaniasis (CL) has been registrated in all Federate Units, including Pará State. Lutzomyia (Lutzomyia) longipalpis is considered to be the main vector of Leishmania (Leishmania) infantum chagasi, the etiological agent of AVL while Lutzomyia (Nyssomyia) antunesi is the proven vector of Leishmania (Viannia) lindenbergi suspected of causing CL. The specificity of the parasitesandflies interaction has to do with leishmania surface molecules as lipophosphoglycan (LPG) which is recognized by receptors of midgut epithelium. However, in permissive sand flies as L.(L.) longipalpis, the interaction is mediated by expressions of glycoproteins containing N-acetylgalactosamine (GalNAc) of epithelial insect midgut which probably acts as recognition sites by molecules lectin-like which are present on the Leishmania surface. The aim of this work was to compare the midgut expression of glycoprotein containing GalNAc of L. (L.) longipalpis from a close colony to the glycoprotein expression of field samples of L. (L.) longipalpis and L. (N.) antunesi of Cametá Municipality, Pará State. Phlebotomines were dissected into 4% paraformaldehyde and each individual midgut was sectioned longitudinally; then incubated with FITC conjugated Helix pomatia lectin (HPA-FITC) and Concanavalin A (Con-A-FITC) which are specific for GalNAc and manose residues, respectively. Lysates of seven midguts of each phlebotomine species from colony and field were analyzed by SDSPAGE followed by western blotting with HPA and Con-A for detecting glycoprotein containing GalNAc and manose. In both colony and field samples, HPA-FITC and Con-A-FITC showed positive reactivity suggesting the presence of both GalNAc and manose. SDS-PAGE followed by western blotting showed that midguts lysates were GalNAc- and manose-positive for displaying peptides ranging from 35 to 75 kDa and 51 kDa that bind to HPA and Con-A, respectively. The results indicated that glycoprotein containing GalNAc occurs in both species from areas endemic for AVL and CL of Cametá municipality. Supported by CAPES, CNPq/UFPa, CNPq/MCT/CT-INFRA/CTPETRO (Process number 620179/2008), MCT/CNPq/FNDCT/PROCAD-NF CAPES/FAPERJ and IEC/SVS/MS. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 109-P Leishmania - Sand fly Interactions ID 110-P Leishmania - Sand fly Interactions COMPARATIVE ANALYSIS BETWEEN CANINE VISCERAL LEISHMANIASIS AND SANDFLY PRESENCE IN THE ENDEMIC MUNICIPALITIES SURVEY OF SANDFLY VECTORS IN AREAS S U R R O U N D I N G P E D R A B R A N C A M A S S I F, MUNICIPALITY OF RIO DE JANEIRO, BRAZIL Denise Maria Bussoni Bertollo, Regiane de Cassia Hilário Castilho, Márcia Maria Costa Nunes Soares, Juliana Kindler Figueiredo, Brígida Helena da Silva Meneghello; Roberto Mitsuyoshi Hiramoto; José Eduardo Tolezano Quezia Nunes Peres Dias, Marcos Barbosa de Souza, Shênia Patrícia Corrêa Novo, Antônio de Medeiros Meira e Cesar do Santo Ponte Instituto Adolfo Lutz 114 Fundação Oswaldo cruz, Fiocruz / Escola Nacional de Saúde Pública Sérgio Arouca, ENSP. [email protected] [email protected] Keywords: Leishmaniasis, Sandfly, entomological investigations Visceral Leishmaniasis (VL) is an infectious disease considered a zoonosis that infects humans as incidental hosts. VL has experienced major changes in the pattern of transmission, initially restricted to rural and peri-urban environments and, more recently, reaching important urban centers. The trypanosomatid protozoan species Leishmania infantum (chagasi) the etiological agent that causes VL, being transmitted by the hematophagous sandfly vector Lutzomyia longipalpis. The objective of this study was to correlate the occurrence of Canine Visceral Leishmaniasis (CVL) and the presence of vectors in municipalities in the northwestern region of São Paulo State, in the period 2008-2014. During the period, 38 municipalities were investigated, in 11 (28.9%) both infected dogs and sandfly vectors were recorded. In 25 (65.7%) only vectors were found and recently 2 human cases were diagnosed. Adults sandflies were predominant in peridomestic environments during entomological investigations in the municipalities with vector. Such environments were characterized by the presence of pets and appropriate habitats for breeding of L. longipalpis in urbanized area. Actions to reduce the probable sites of vector propagation have been performed by means of environmental management in these municipalities. Climatic factors may determine the epidemiology of VL and the adaptation and gradual migration of vectors to urban regions. Factors such as temperature, humidity, rainfall and presence of sandflies must be studied and understood in order to reduce the spread of vector populations and consequent increase in disease transmission. Keywords: Leishmaniasis, Sandfly fauna, Pedra Branca Massif, Municipality of Rio de Janeiro Introduction: Several sand fly species have been registered with natural infection with Leishmania spp. In Brazil, American cutaneous leishmaniasis (ACL) is worthy of attention due to its high incidence and risk of deformities in humans. The incriminated vectors of ACL are Nyssomyia intermedia, Migonemyia migonei, Nyssomyia whitmani, Bichromomyia falviscutellata, Nyssomyia umbratilis and Psychodopygus wellcomei. American visceral leishmaniasis (AVL) has been expanding rapidly in urban areas, having as its principal vectors Lutzomyia longipalpis and Lutzomyia cruzi. Materials and Methods: This study was conducted in 31 endemic areas in the surroundings of Pedra Branca Massif, to the west of the city of Rio de Janeiro, Brazil: A. Vasconcelos, Barra de Guaratiba, Serra dos Caboclos, Cabuçu, Cachamorra, Camorim, Carapiá, Colônia Juliano Moreira, Estrada do Magarça, Estrada do Monteiro, Estrada das Taxas, Estrada dos Teixeiras, Grota Funda, Grumari, Guandu do Sena, Ilha de Guaratiba, Jardim Novo Realengo, Lameirão Pequeno, Maciço do Mendanha, Pau da Fome, Serra de Piabas, Rio Bonito, Santa Maria, Serra do Barata, Serra do Bangu, Serra do Rio da Prata, Serra de Santíssimo, Serra do Viegas, Vargem Grande, Vargem Pequena and Vila Kennedy. Sand fly collections were conducted during two years (2003-2005) in peridomestic habitats using CDC light traps, targeting the sand fly habitats in these regions. The collections were performed from 6 pm to 7 am the following day and specimens were analyzed in the laboratory and identified to species level. Results: A total of 46,971 specimens belonging to 13 species were collected. N. intermedia, M. migonei and L. longipalpis were the most prevalent species with a total of 40,449; 4,072 and 1,750 specimens, respectively. The months with highest sand fly density were June, July, September and March. Discussion and Conclusion: The high density of the three most common species can be explained by the presence of hen houses in these localities, corroborating the literature on studies conducted in different localities. This survey enabled us to discriminate and quantify the species by regions, contributing to eco-epidemiological data about vector, once the risk of transmission of the parasite depends on the existence of a large number of vector species, and also the emergence of disease previously restricted to the sylvatic species. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 111-P Physiology and Colonization ID 112-P Physiology & Colonization ROTATION OF THE EXTERNAL GENITALIA IN MALE PHLEBOTOMINE SANDFLIES (DIPTERA, PSYCHODIDAE) IN LABORATORY CONDITIONS AND IN CAPTURED SPECIMENS IN ALGARVE, PORTUGAL SOME FIELD AND LABORATORY OBSERVATIONS ON THE BIOLOGY OF NYSSOMYIA ANTUNESI (DIPTERA: PSYCHODIADAE), A SUSPECTED VECTOR OF LEISHMANIA (VIANNIA) LINDENBERGI IN AMAZONIAN BRAZIL Ferrolho J.1, Gomes J.1, Alves-Pires C2, Cristóvão J.M.1, Maia C.1,3, Campino L.1,2, Afonso M.O.1,2 Thiago Vasconcelos dos Santos1, Marliane Batista Campos1, José Aprígio Nunes Lima1, Roberto Carlos Feitosa Brandão1, Fábio Márcio Medeiros da Silva1, Maria Sueli Barros Pinheiro1, Luciene Aranha da Silva Santos1, Edna De Freitas Leão1, Iorlando da Rocha Barata1, Fernando Tobias Silveira1 1 UEI Parasitologia Médica, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa (IHMT-UNL), Rua da Junqueira 100, 1349-008 Lisboa, Portugal; 2 UPMM/FCT, IHMT-UNL; 3CMDT/FCT, IHMT-UNL [email protected] 115 1Laboratório de Leishmanioses “Prof. Dr. Ralph Lainson”, Seção de Parasitologia, Instituto Evandro Chagas (SVS/ MS), Ananindeua, Pará, Brazil; [email protected] Keywords: Phlebotomine sandflies, genitalia, Portugal Phlebotomine sandfly females are responsible for the transmission of several pathogens, such as Leishmania spp., affecting humans and animals in the Old and New World. Recently, ecological and climatic changes have been associated with an increase of vectorial density and leishmaniasis expansion. Phlebotomus perniciosus and P. ariasi are the proven vectors of L. infantum in Portugal. Phlebotomine males eclose from the pupae with un-rotated external genitalia. During the initial 16-24 hours of adult life time, the external genitalia has to undertake a complete rotation on the longitudinal axis through 180° to assume a mature position. Recording the physical characteristic of males with unpartially or completely rotated external genitalia can help to predict the breeding sites and the proximity of adult movements. In the present study, we firstly investigated the timing of complete rotation of the male external genitalia in laboratory conditions. P. perniciosus males were used as a model and were obtained from a previously established colony at the IHMT-UNL and maintained with optimal rearing conditions throughout the assay. After eclosion, males were separated and observations were carried out at 24 and 48 hours. At each sampling time point, external genitalia were observed under a stereomicroscopic, either in vivo or after preservation in 90% ethanol. Secondly, we identified morphologically the sandfly male species captured from May to November 2013 in the Sotavento and Barlavento Algarvio regions, Portugal, in different biotopes types (domestic and peridomestic) using CDC miniature light and sticky oil paper traps. Finally, we characterized the external genitalia rotation in the captured males and investigated the relation between the species and types of traps, periods of capture and natural biotopes/collecting places. In this work, we report the timing for external genitalia to complete rotation under laboratory conditions in P. perniciosus males, so far the only confirmed vector of L. infantum in Algarve, and based upon this model, we related the proximity of the captured sandfly species with their breeding sites in the selected Algarve regions. More information on sandfly breeding sites will facilitate their control by source reduction and host protection, especially in urban and periurban areas. This work was partially supported by EDENext (FP7-INCO-CT-2010-261504). C. Maia (SFRH/BPD/44082/2008) holds a fellowship from FCT, MCTES, Portugal Keywords: Nyssomyia antunesi, Leishmania (Viannia) lindenbergi, cutaneous leishmaniasis. In the Amazon region, the phlebotomine sand fly Nyssomyia antunesi Coutinho, 1939 is recognized to be the main suspected vector of cutaneous leishmaniasis (CL) caused by Leishmania (Viannia) lindenbergi Silveira et al. 2002, with basis on the anthropophilic behavior and high frequency in endemic areas under the occurrence of the disease. Despite the epidemiological relevance, little is known regarding the biology of this sand fly. Thus, this study aimed to describe some field and laboratory observations of Ny. antunesi in order to provide further evidences on the ecology of CL in this region. During the Amazonian dry season (September-October/2013), an entomological survey was conducted in a forest fragment located nearby Maguari River margin (1º20'21''S 48º22'13''W) in Ananindeua municipality, Pará State, Brazil. This location was chosen based on a previous human case of CL attended at the Laboratório de Leishmanioses “Prof Dr. Ralph Lainson”, Instituto Evandro Chagas, Ananindeua, Brazil. Collects were performed for 15 nights using CDC light traps at 1,5m ground (n=6) and 20m at canopy (n=2). The collected sand flies were identified and the females were dissected to search flagellate infection. In addition, some of collected sand flies were kept alive to be fed on hamster and the matrixes were prepared individually or in groups of 50 individuals for oviposition. Immature emerged stages were maintained with a diet based on a mixture of triturated substrate and powdered beef liver (Difco). A total of 2.414 sand flies were collected and identified, belonging to 13 species. Ny. antunesi accounted 83,7% of collected at 1,5m and 91,3% at 20m. Two dissected females of this species were found harboring flagellates morphologically distinguishable from Leishmania spp. Out of approximately 2000 non-identified individuals kept alive, 70% fed on hamster. Individualized females did not succeed for oviposition. Two groups of 50 females generated F1 and achieved until adult form in approximately 52 days with the following duration average of immature stages: egg (10 days), L1 (9 days), L2 (11 days), L3 (9 days), L4 (7 days), pupa (6). All the adults emerged were confirmed as Ny. antunesi. However, the F2 did not continued successfully. These results together with previous data corroborate to indicate Ny. antunesi as potentially involved on the transmission of L. (V.) lindenbergi in the Amazon region. Under laboratory conditions, immature stages of Ny. antunesi seem to be well adapted for colonization, however, survival of adult form should be more carefully analyzed. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 113-P Physiology & Colonization ID 114-P Physiology & Colonization ATTRACTIVENESS OF DIFFERENT DIETS FOR NYSSOMYIA NEIVAI (DIPTERA: PSYCHODIDAE) LARVAE EFFECT OF NYSSOMYIA NEIVAI (DIPTERA: PSYCHODIDAE) DENSITY ON OVIPOSITION 1 2 Thais Marchi Goulart ; Vicente Estevam Machado ; Mara Cristina Pinto2 1 Instituto de Biologia, Departamento de Biologia Animal-UNICAMP, Brazil; Faculdade de Ciências Farmacêuticas-UNESP, Brazil 116 Thais Marchi Goulart1; Vicente Estevam Machado2; Mara Cristina Pinto2 1 Instituto de Biologia; Departamento de Biologia Animal-UNICAMP; 2Faculdade de Ciências Farmacêuticas-UNESP 2 [email protected] [email protected] Keywords: colony, density, oviposition Keywords: Sand fly colony, immature stages, attractiveness of sand fly diet The necessity of improving techniques for sand flies colonization is an important demand for further studies on vector biology and its interaction with Leishmania spp. The crucial limitation for rearing these insects is the high mortality during immature phases. Although there are some studies focused on diets for sand fly larvae, this aspect is still an issue for sand fly colonies. The aim of this research was to evaluate the attraction of immature stages of Nyssomyia neivai for different diets. Sand flies were collected in Santa Eudóxia, SP - Brazil (edges of Mogi Guaçu river) using automatic light traps and manual aspiration. At the laboratory, sand flies were maintained in cages covered with "voil" (30x30x30cm) at 26 ± 1°C, 80–90% humidity, 12:12 (L:D) photoperiod with access to a piece of cotton soaked in 30% sucrose solution. The non-bloodfed field-collected females were exposed to mouse as blood source. The bloodfed females were transferred to an oviposition chamber. Fifty eggs of N. neivai were placed in the center of a 150mL polystyrene container filled with 2 cm of plaster of Paris. The plaster of Paris was divided in four quadrants and over each one a specific diet was added: diet 1 (rabbit feces, rabbit food, fish food and soil), diet 2 (rabbit feces and soil), diet 3 (rabbit feces, soil and fish food) and diet 4 (soil and mud). The number of larvae on each quadrant was observed daily. The containers were maintained in the dark at 26 ± 1°C, 80–90% humidity. The experiment was replicated four times. The results were analyzed by analysis of variance (ANOVA) and Tukey test. According to the result, the most attractive diet was diet 1, followed by diet 3. Both diets contain fish food. Differences were considered significant among the four diets (p<0.0001) but with no significant difference were verified between diets 1 and 3 (p>0.05) or 2 and 4 (p>0.05). The presence of fish food appears to be important on the attractiveness of the larvae stages. This information is interesting for further studies on sand flies larvae attractiveness. The maintenance of sand flies colonies is a meticulous activity and each species seems to have special requirements. Nyssomyia neivai is an important vector of cutaneous leishmaniasis in South America and data about its colonization are scarce. The aim of this study was to evaluate females of N. neivai density effects on oviposition. Sand flies were collected in Santa Eudóxia, SP - Brazil from 6 p.m. to 11 p.m. using Castro aspirators. At the laboratory sand flies were maintained in holding cages at 26 ± 1°C, 80-90% humidity, 12:12 (L:D) photoperiod with access to a piece of cotton soaked in 30% sucrose solution. The non-bloodfed field-collected females were exposed to a mouse as blood source. Bloodfed females were selected and transferred in groups with equal numbers of males to 250mL polystyrene oviposition chambers. The cover of the chamber was replaced with “voile” and the bottom filled with 2cm of plaster of Paris as a substrate for oviposition. The sand fly couples were placed in seven groups: 1, 5, 10, 15, 20, 25 and 30. The experiment was replicated four times. The chambers were observed daily to check the humidity, change of sucrose solution, mortality of gravid females and presence of eggs. After the egg counting, dead sand flies were removed and identified. The chambers were maintained in the dark at 26 ± 1°C, 80-90% humidity. The results were analyzed by analysis of variance (ANOVA) and Tukey test. As result, all the females were identified as N. neivai. The mean of eggs per female was highest when sand flies were in groups than individualized, but the differences were not statistically detected. The range of the means varied from 37.3 (± 5.3) eggs in chambers with only one female to 56.5 (± 3.9) eggs in chambers with 25 females. When the number of couples in the chamber is highest than 20, the females lay eggs on the top, “voile” and wall of the chamber and due to this fact, these eggs dry and die. The results obtained with N. neivai using groups of females indicated that there is an increase on oviposition when females are in groups, but considering all the conditions for the process, the ideal number of couples for this species in the container of 250mL is 20. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 115-P Physiology & Colonization ID 116-P Other Topics ROLE OF AQUAPORINS IN THE PHYSIOLOGY OF LEISHMANIA VECTOR LUTZOMYIA LONGIPALPIS LEISHMANIASIS VIRTUAL LABORATORY (LVL) - A NEW TOOL FOR THE STUDY OF AMERICAN SAND FLIES (DIPTERA, PSYCHODIDADE, PHLEBOTOMINAE) Rita Mukhopadhyay1, Goutam Mandal1, Jose Orta1, Srotoswoti 1 Mandal , Claudio Meneses2, Hamide Aslan Suau2, Eric Beitz3, Jesus Valenzuela2, Marcelo Ramalho-Ortigao4, Shaden Kamhawi2 1 Department of Cellular Biology and Pharmacology, Herbert Wertheim College of Medicine, Florida International University, Miami, USA; 2Vector Biology, National Institute of Allergy and Infectious Diseases, Baltimore, USA; 3Dept. of Pharmaceutical and Medicinal Chemistry Pharmaceutical Institute, University of Kiel, Germany; 4 Department of Entomology, Kansas State University, Kansas, USA. [email protected] Keywords: Leishmania, aquaporins, Lutzomyia, sand fly Introduction: Aquaporins (AQP) are the route of entry of water and uncharged solutes including gases (carbon dioxide and ammonia) into cells from bacteria to humans. They play highly significant roles from cell migration in mammals to fecundity in Drosophila. Female phlebotomine sand flies are vectors of Leishmania.sp worldwide. Sand flies survive in widely variant climatic conditions of extremely humid tropics to the deserts of the Middle East. Upon blood meal ingestion, a large number of events are induced, including digestion, metabolism, diuresis and ultimately oogenesis. Sand flies also ingest plant sap or aphid and coccid honeydew rich in sucrose between blood meals. Therefore, their gut has to deal with huge volumes of fluids/water and a wide range of osmolarity. On the other hand, in nature they oviposit on substrates ranging from decaying organic matter to animals feces. How do they home in to these various sources of feed either for themselves or for their young? There is a substantial gap in understanding the underlying molecular mechanisms of these physiological processes of the sand flies. We hypothesize that aquaporins (AQPs) are at the center of these primary sand fly survival activities. Yet AQPs have not been reported from this highly important disease vector. Methods: We cloned five Lutzomyia longipalpis (LULO) AQPs by RACE-PCR and expressed them in Xenopus oocytes. Additionally, various tissues under different conditions were isolated from the flies, and different AQP status was determined by qPCR. Results: All AQPs facilitate water uptake in Xenopus oocytes. Modulation of AQPs was also observed based on the feeding status, tissue specificity and Leishmania infection. LULO-AQP1-4 are also transcriptionally active in antennae and brain. Discussion: AQPs are involved in major physiological processes of Lutzomyia. We also propose that they are involved in homing behavior which is novel in insect research. 117 Felipe Dutra Rêgo; Israel de Souza Pinto, Caroline Amaral Machado, Gabriel Eduardo Melim Ferreira, Elisa Cupollilo, Paloma Helena Fernandes Shimabukuro Centro de Referência Nacional e Internacional para Flebotomíneos/Centro de Pesquisas René Rachou/FIOCRUZ [email protected] Keywords: Barcode, Cloud computing, Phlebotominae Cloud computing has becoming increasingly popular among different kinds of internet services available to users, and it involves platforms or applications that offer services remotely through the web. Cloud-based services can be central to scientific advancements, challenging the sustainability of an approach to distributed computing rapidly being taken over by events and increased demands from user needs in the past 15 years. The EU-Brazil Cloud Connect (EUBrazilCC) project is built on a close collaboration between European and Brazilian institutions working towards providing a user-centric, real test bench for European and Brazilian research communities to test the execution of scientific applications. This project is being developed with three scientific use cases covering multidisciplinary areas, such as biodiversity, epidemiology, public health, and climate change. These scientific scenarios all require complex workflow pipelines and access to huge datasets. One of the scientific use cases is the Leishmaniasis Virtual Laboratory (LVL), a collaboration between the Instituto de Salud Carlos III (WHO-CCL Leishmania Collection) in Spain and FIOCRUZ in Brazil with the collections of Leishmania (CLIOC) and sand flies (COLFLEB). The aim of this work is to present the LVL use case to the sand fly research community and to provide a pipeline to perform molecular analyses of sand flies. At COLFLEB, our aim is to study the phylogenetic relationships of American sand flies, including the main vectors of Leishmania in Brazil (Nyssomyia intermedia, N. neivai, N. whitmani, N. umbratilis, Bichromomyia flaviscutellata, Psychodopygus spp., and Lutzomyia longipalpis). Using cytochrome oxidase subunit I (coI), our pipeline will work with uploaded sequence data from sand flies generated by user or downloaded from public databases (e.g. Barcode of Life Database - BOLD or GenBank). Analysis of data through the pipeline will include: DNA sequence editing and alignment, maximum likelihood analysis, Bayesian inference analysis and tree editing. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 117-P Other Topics ID 118-P Other Topics IMMUNOLOGICAL STUDIES ON THE CUTANEOUS LEISHMANIASIS IN AL-KHARJ REGION, SAUDI ARABIA SANDFLIES AND POPULAR KNOWLEDGE OF LEISHMANIASIS TRANSMISSION IN A PERI-URBAN AREA OF THE MUNICIPALITY OF SÃO GONÇALO DO AMARANTE, BRAZIL Mohammad S. Al-Khalifa; Ibraheem M Al-Hazza and M. Y. Lubbad Zoology Department, College of Science, P. O.Box 2455, King Saud University, Riyadh 11451, Saudi Arabia. [email protected] Keywords: Cutaneous Leishmaniasis, Phlebotomus, Saudi Arabia, Zoonosis. Present study is an attempt to determine the nature of immunological cells and the cytokine they produce in zoonotic cutaneous leishmaniasis (ZCL) patients in an endemic zone, Al-Kharj area, southeast of Riyadh Saudi Arabia. ELISA and the short-term culture were the techniques used for this purpose. ELISA determines the level of cytokines in the peripheral blood in ZCL patients. There were no variations in the levels of IL-4 in the blood of ZCL patients before and after treatment by tropical application of Pentostam. However, level of cytokine IL-6 and IL-10 and TNF-α have significantly increased in the patient's blood. Short-term cultures were used to study the level of intra cellular cytokine and it observed that level of intra cellular IL-2 has significantly increased before the treatment compared to the same patient after the Pentostam treatment. However, its level increased when examined in patients after the three months treatment and there were no variation in the levels of IL-4 in ZCL patients, before and after treatment. 118 Maria de Lima Alves, Maria de Fátima Freire de Melo Ximenes, Marcos Paulo Gomes Pinheiro, Tamy Elicia da Silva Lopes, João Batista da Silva Junior Universidade Federal do Rio Grandedo Norte - UFRN. [email protected] Keywords: Popular knowledge, Phlebotomine sandflies, health education The peri-urban areas of cities are of particular interest to researchers, especially in the field of public health, due to the socioeconomic, environmental, sanitary and cultural characteristics of these regions, which are unfavorable for public and collective health. In Rio Grande do Norte, leishmaniases occur in different geographic regions of the state both separately and concomitantly. The number of cases of Visceral Leishmaniasis (VL) is higher than that of American Cutaneous Leishmaniasis (ACL). The aim was to characterize the community as to social, sanitary and environmental aspects as well as the knowledge of residents regarding the transmission cycle of leishmaniases in the Guanduba community located in the peri-urban area of the municipality of São Gonçalo do Amarante. The methodology consisted of applying a questionnaire containing twenty semi-structured questions on epidemiological, sanitary and environmental aspects involved in the transmission cycle of leishmaniasis to 115 families enrolled in the Family Health Program (PSF), Ministry of Health, in July 2013. The results demonstrate that the Guanduba community suffers from socioeconomic problems, the absence of urban and sanitary infrastructure as well as a number of environmental problems. With respect to the disease process, 66% of those interviewed did not know how leishmaniasis was contracted, 52% were unaware of how it was transmitted, 96% did not know how to prevent it and 95% did not know the name of the transmitting insect of leishmaniases. In the comunity was held on systematic capture of sandflies with CDC light traps, which demonstrate the occurrence of species incriminated as vectors of LV and LTA. The catches indicate the presence of Evandromyia lenti 84%, Evandromyia evandroi 8%, Lutzomyia longipalpis 6%, Sciopemyia sordellii 1%, Evandromyia walkeri 0,5% e Nyssomyia intermedia 0,5%. It is concluded that the level of knowledge regarding epidemiological cycles is weak and inadequate for prevention measures, to the detriment of the accompaniment of these families by the PSF in this community, where sanitary conditions, environmental and entomological will increase the risk of disease transmission, as evidence the presence of vector species of leishmaniasis. Thus, it is recommended that educational measures being implemented by the Family Health Strategy in partnership with other public institutions. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 119 ID 119-P Other Topics ID 120-P Other Topics BOOKLET EDUCATIONAL IN COMIC STRIP ABOUT SANDFLIES AND THE LEISHMANIASIS IN THE STATE OF RIO GRANDE DO NORTE, BRAZIL Maria de Lima Alves, Maria de Fátima Freire de Melo Ximenes, Magnólia Florêncio Fernandes de Araújo, Marcos Paulo Gomes Pinheiro, Tamy Elicia da Silva Lopes, João Batista da Silva Junior, Ileane Gurgel Mendes Universidade Federal Do Rio Grande Do Norte - UFRN CORDEL FOLK LITERATURE AS A TOOL FOR PREVENTING LEISHMANIASIS IN BRAZIL Marcos Paulo Gomes Pinheiro, Maria de Fátima Freire de Melo Ximenes, Maria Paula de Melo Pereira Pinheiro, Maria de Lima Alves Universidade Federal do Rio Grande do Norte [email protected] [email protected] Keywords: Cordel Literature, Leishmaniases, Phlebotomine sandflies, Education. Keywords: Phlebotomine sandflies, Educational comic strip booklet, health education. Leishmaniases, a set of diseases transmitted by phlebotomine sandflies, are one of the greatest challenges faced by public health due to the lack of mechanisms, technology, drugs, vaccines and effective educational measures to control them. The expansion of Visceral Leishmaniasis (VL) and American Cutaneous Leishmaniasis (ACL) in Brazil is associated to the adaptation of the vector to environments modified by anthropic action and the precarious socioeconomic and sanitary conditions of the population. In Rio Grande do Norte visceral and cutaneous leishmaniases are expanding. A number of factors contribute to this situation, in addition to aspects related to the species and abundance of sandflies in this region. In the year 2012 were peformed systematic captures of sandflies with CDC light traps in two counties in the state of Rio Grande do Norte, where showed the occurrence of species incriminated as vectors of LV and LTA. In the first countie this state, São Gonçalo do Amarante, was found the presence of Evandromyia lenti (84%), Evandromyia evandroi (8%), Lutzomyia longipalpis (6%), Sciopemyia sordellii (1%), Evandromyia walkeri (0,5%) and Nyssomyia intermedia (0,5%). Already in the countie of Assu was evidenced lower variation of species predominating Lutzomyia longipalpis (98%), Evandromyia evandroi (1,62%), Evandromyia lenti (0,11%) and Micropygomyia trinidadensis (0,03%). In this sense, the knowledge of sandfly species contributes to the understanding of the cycle of transmitting the disease being essential to its control. The use of didactic instruments in schools and the community is an excellent health education tool for disseminating knowledge and controlling the disease. The aim was to create a 28-page educational comic strip booklet containing information on the cycle of the vector, transmission of the parasite, its life cycle on different hosts and the forms of leishmaniasis manifestation. The booklet will be distributed in elementary and secondary schools as well as to other groups during environmental health education campaigns aimed at providing knowledge and recognition of sandflies to the population. The entertaining nature of comic strips contributes to involving the reader through the words and images of well-known personalities who represent and legitimize the scientific discourse and to disease prevention, health promotion and engaging the population, as well as their participation in issues related to health and quality of life. Cordel literature (literally “string literature”) is considered a form of popular art unique to Brazil, published as rhyming verses, produced in small inexpensively printed booklets, called cordel because they were originally hung from strings. This type of literature, in addition to entertaining, informal and pleasurable, can be easily introduced to social settings. It consists of folk novels and poems that can be easily inserted into an educational context to popularize science. Northeastern Brazil has the highest incidence of visceral leishmaniasis, affecting primarily the more underprivileged social classes. The study aims at preventing leishmaniasis via cordel literature, given its acceptance and entertaining aspect, with a focus on the transmission, symptoms and prevention of leishmaniases. In this respect, a booklet entitled “ A drinker of blood in the land of the mandacaru cactus – the fight against Kalazar” was manufactured to be used in environmental and sanitary education regarding leishmaniases, with populations residing near three environment protection areas in the state of Rio Grande do Norte, in Northeastern Brazil, two in the Caatinga biome in the municipalities of Assú and Serra Negra do Norte and one in the Atlantic Forest, located in the municipality of Nísia Floresta. Phlebotomine sandly collections were conducted in the aforementioned areas, using CDC traps. Vector species were captured, highlighting the need for educational measures. Collections indicate the presence of Psychodopygus wellcomei, a vector species of cutaneous leishmaniasis in the Atlantic Forest area, and that Lutzomyia longipalpis, a vector of visceral leishmaniasis, is the most abundant in anthropized environments, as well as in the preserved portion of the Caatinga. The most collected species in the Atlantic Forest area were Lutzomyia walkeri (76.6%), Evandromyia evandroi (16.5%) and P. wellcomei (1.8%), and in the Caatinga L. longipalpis (98%) and L. evandroi (1.6%). Given the lack of information on the topic, educational initiatives were carried out in a school setting. The interest of individuals in reading the booklets can be readily observed and we can therefore use them to increase community awareness, since this type of literature is easily accessed, very inexpensive, in addition to reaching all age groups, and social and educational levels, making it an easy, low-cost way to disseminate information on an tissue of great importance to public health. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 121-P Other Topics ID 122-P Other Topics S U R V E Y O F A M E R I C A N T E G U M E N TA R Y L E I S H M A N I A S I S I N T H E N O RT H E A S T O F ARGENTINA DURING 2011-2014 CANINE VISCERAL LEISHMANIASIS. IMPORTED CASES IN URUGUAY, 2010-2014 Lozano Alejandra1; Basmadjián Yester2; Vitale Edgardo1; Satragno Dinora2; Canneva Bruno2; Verger Lorenzo1; Tort Cecilia2; Viera Ana2; Romero Selva2; Ríos Cristina3 and Lagarmilla Patricia3 María Josefa Felisa Rea, Carlos Edgardo Borda Centro Nacional de Parasitología y Enfermedades Tropicales (Cenpetrop), Facultad de Medicina, Universidad Nacional del Nordeste [email protected] Keywords: American tegumentary leishmaniasis Argentina 120 northeast 1 Área de Medicina Preventiva y Epidemiología, Facultad de Veterinaria, Universidad de la República, Montevideo. Uruguay. Alberto Lasplaces 1550. 2Departamento de Parasitología y Micología Médica. Instituto de Higiene, Facultad de Medicina, Universidad de la República. Montevideo. Uruguay. Avda. Alfredo Navarro 3051, Montevideo, Uruguay. 3Área de Salud Pública Veterinaria, Facultad de Veterinaria, Universidad de la República. Montevideo. Uruguay. [email protected] Keywords: Leishmaniasis visceral, Uruguay, dogs American tegumentary leishmaniasis (ATL) is an endemic disease, in some parts of Argentina. Corrientes province, located in the Northeast region, report many leishmaniasis cases with epidemic outbreaks recorded in three municipalities. One hundred forty seven human cases of ATL were diagnosed from 1987-2010 in the CENPETROP. Leishmania (Viannia) guyanensis, Le. (V.) braziliensis and Le. (Le.) amazonensis have been isolated from human cases.The purpose of this study was to describe seven new cases of disease from 20102014, and to identify potential sandfly vectors in peridomicile of two patients. Four patients were men and three women. Age range was between 28-82 years old. From the province of Corrientes were five patients(two from Corrientes city and three from the Departments of Lavalle, San Luis del Palmar and Bella Vista), one from the province of Formosa(Fontana)and another from the Paraguay Republic(Pilar).Ulcers were present in four patients and the legs were the most affected parts of the body.Two patients presented only mucosal involvement (nasal and oral) and in one patient with two skin ulcers, mucosal compromise was observed one year later of the primary lesion.The Montenegro skin test was positive in all patients and the average size was 9.3mm and IM Glucantime® was administered. No relapse of infection was recorded in cured patients, but in two women with a complete clinical cures both suffered traumatism in the same place of the lesion scar within four or five years later, thus they relapsed and a new ulcer was formed. A 64 yearold male was erroneously diagnosed of epithelium cancer, thus he received a transplantation of tissue. Between 9 and 12 months later this patient had relapse and a new ulcer was formed in the same place of transplant and with metastasis in the oral mucous. Using light traps and sticky-paper traps, sand flies were collected in the houses of two patients from Corrientes and Bella Vista. In Corrientes three species: Lutzomyia (Nyssomia) nievai, Lu. (Pintomyia) shannoni and Lu. (migonei) migonei were collected. The first species was dominant (86%). In Bella Vista 15 Lu. (Ny.) nievai were collected. In both, Lu. (Ny.) nievai was the most common species and therefore suspected to play the main role in ATL transmission. Further studies are needed to verify the role of some species of sand flies as vectors of leishmaniasis in the area. The occurrence of these cases demonstrates the persistence endemic of the disease. Visceral Leishmaniasis (VL) is a parasitic zoonotic disease caused by the protozoan Leishmania infantum and transmitted by the vector Lutzomyia longipalpis. The disease affects several species of mammals and the dog is considered its main reservoir. In the year 2000 took place the emergence of this disease in the Southern Cone of the Americas, being the urban cycle the main form of transmission. In this cycle, the dog plays a major role since, apart from being the parasite´s main host, is in a close relation with the human being. At the present date, VL is an exotic disease in Uruguay, since no autochthonous cases were described in humans or dogs. L. longipalpis was found for the first time in Uruguay in the year 2010, in the northern cities of Salto and Bella Unión. Since then, the whole country was classified as vulnerable for the emergence and dissemination of VL. The current situation is worsened by the occurrence of human and canine cases in border locations and the entry of infected dogs from endemic areas. These dogs enter the country accompanying immigrant families or people returning to the country after spending years abroad. In the present work, a description of the imported canine cases that entered the country is performed. Between the years 2010 and 2014 six cases were detected, five already came with a diagnosis of VL and one was diagnosed in a veterinary practice shortly after its arrival. Three dogs came from Europe: two from Spain and one from France. The remaining three dogs came from Paraguay. Once in the country, the owners of the infected dogs took up residence in the departments of Maldonado, Canelones and Montevideo. On the follow up of the cases: two dogs died from the disease, two were euthanized and one (whose owner refused euthanasia) is still living in Montevideo. Contact was lost with the remaining two dogs. Given the current epidemiologic situation of the country, it is essential to have a clear policy on how to handle the dogs infected. It is also strongly recommended harden the controls on the dogs coming from areas where VL is endemic and avoid that any infected dog reach an area where L. longipalpis is present. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 123-P Ecology & Epidemiology ID 124-P Ecology & Epidemiology OCCURRENCE OF LUTZOMYIA LONGIPALPIS AND S PAT I A L D I S T R I B U T I O N O F V I S C E R A L LEISHMANIASIS IN FLORIANO, PIAUÍ, BRAZIL: PRELIMINARY DATA USE OF REMOTE SENSING IN THE ANALYSIS OF POPULATIONS OF LUTZOMYIA LONGIPALPIS COMPLEX Anárya Teresa de Freitas Rocha¹, Simone Melo Silva¹, Maria Regiane Araújo Soares²; Daniel Costa Fortier²; Diogo Brunno e Silva Barbosa2; Carlos Henrique Nery Costa2 1 Biologist, Universidade Federal do Piauí; 2Researcher. Universidade Federal do Piauí. [email protected] Keywords: visceral leishmaniasis, georeferencing, hotspots Visceral leishmaniasis (VL), caused by Leishmania infantum and transmitted by Lutzomyia longipalpis, was first recorded in Brazil in the beginning of the 20th century as a rural disease and has expanded to urban centers since the 1980's. This work describes the occurrence of Lu. longipalpis and spatial distribution of human cases of VL in the municipality of Floriano, Piauí, Brazil, as well as discusses the influence of environmental variables on their notification. Data on notification were collected from the Sistema de Informação de Agravos de Notificação (SINAN) and the Centro de Controle de Zoonoses (CCZ), and includes the years 2005 and 2007 to 2013. VL cases were georeferenced using the Global Positioning System, followed by the preparation of a distribution map (Google Earth software). Environmental variables (temperature, relative humidity and rainfall) were obtained by Instituto Nacional de Meteorologia. Data on sand flies were obtained in 2014, through collections performed in areas of higher occurrence of VL based on the previous information provided by SINAN and CCZ. Collections were quarterly carried out in animal shelters and residences using light traps. During the study period, 66 cases were reported, with the largest number of records in 2009 (n=17). Among the districts, “Tiberão” detained most of the records (n=6), followed by “São Borja” (n=5) and “Viazul” (n=5), all located in the periurban area of the city. At “Tiberão”, it was observed that all notifications (n=6) were composed by two people in the same household in three consecutive years (2008 to 2010). In this district, 496 specimens of Lu. longipalpis (413 ♂ and 83 ♀ ) were collected in the first quarter of 2014, 85% of which in peridomestic environment. Only 38 cases were georeferenced and used for the preparation of spatial distribution maps, thus excluding the cases with incomplete information. A reduction in the number of cases in the countryside was observed, in contrast to peripheral districts that represent the expansion area of the city. There was no correlation (Spearman test) among the environmental variables and case reporting (temperature, p=0.28, relative humidity, p=0.61, and rainfall, p=0.69). New technologies are powerful tools for defining risk areas for action and execution plans to control VL. The occurrence of Lu. longipalpis in peripheral urban areas suggests that these areas may provide favorable conditions for the establishment of natural breeding vector. 121 Mirella FC Santos1, 2; Antonio C Paranhos Filho3; Roberto M Gamarra3; Wedson D Fernandes4; Reginaldo P Brazil5; Alessandra G Oliveira1, 2, 6 1 Post Graduate Program in Animal Biology, Federal University of Mato Grosso do Sul, Campo Grande, Brazil. 2Post Graduate Program in Infectious and Parasitic Diseases, Federal University of Mato Grosso do Sul, Campo Grande, Brazil. 3Geotechnology Laboratory, Federal University of Mato Grosso do Sul, Campo Grande, Brazil. 4 Federal University of Great Dourados, Mato Grosso do Sul, Brazil. 5Infectious Diseases Laboratory, Oswaldo Cruz Institute, Oswaldo Cruz Foundation, Rio de Janeiro, Brazil. 6Parasitology Laboratory, Federal University of Mato Grosso do Sul, Campo Grande, Brazil. [email protected] Keywords: Vector, leishmaniases, NDVI, NDWI Visceral Leishmaniasis (VL) is the most severe form of leishmaniases. In Latin America, the VL etiologic agent is the protozoa Leishmania (L.) infantum and its main vector Lutzomyia longipalpis. In Mato Grosso do Sul (MS), there is a second species, Lutzomyia cruzi, incriminated as a transmitter of the protozoa. In recent years, there has been a marked development of studies relating health and environment. Both can be influenced by space position in where they are inserted, interfering in their conditions and characteristics. Thus, the objective of this study was to determine whether environmental factors such as vegetation and the presence of water, assessed by remote sensing, could explain molecular and morphometric differences between populations of Lutzomyia longipalpis complex in MS. For this purpose, the capture points and the urbanized areas relating to seven populations "longipalpis" complex were analyzed: a population of Lu. cruzi (Corumbá) five populations of Lu. longipalpis in MS (Campo Grande Aquidauna Miranda Tres Lagoas and Bonito) and an outgroup of Lu. longipalpis from Estrela de Alagoas (AL). Remote sensing was used for derivation of ecological indices NDVI (Normalized Difference of Vegetation Index) and NDWI (Normalized Difference of Water Index). In order to find out if these environmental variables reflect the molecular and morphometric differences of the populations studied an ANOVA (F test) was performed through BioEstat 5.0 program. In all study sites was possible to observe that the sparse vegetation was predominant which was expected considering that all areas were urbanized. The higher NDVI values were Estrela de Alagoas and Corumbá one with a recent urbanization and the other containing areas with large limitations of occupancy. It is believed that environmental factors may mask patterns of genetic isolation often found in studies of population genetics but in this study these biotic factors in the dimensioned scale would not be exerting selective pressure. It was observed that the Municipality of Tres Lagoas is the most urbanized with lower values of NDVI and NDWI. The results of this study demonstrate that the species Lu. longipalpis is adapted to the conditions and characteristics of the urban environment as it is associated with the occurrence of different humidity conditions and the supply of vegetation. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 125-P Ecology & Epidemiology ID 126-P Ecology & Epidemiology GEOMETRIC MORPHOMETRY OF NORTHEASTERN POPULATIONS OF NYSSOMYIA NEIVAI FROM ARGENTINA SANDFLIES (DIPTERA: PSYCHODIDAE) OF AGUAS CALIENTES, LA CONVENCIÓN, CUSCO, PERÚ a,b a c,d Utgés, María Eugenia ; Fuenzalida Denise ; Parras Matías ; Gould Ignaciob; Casertano Sergioa ; Salomón, Oscar Daniela, c a REDILA Instituto Nacional de Medicina Tropical (INMeT), Ministerio de Salud de la Nación, Puerto Iguazú, Misiones, Argentina; b Centro Nacional de Diagnóstico e Investigación en Endemo-epidemias (CeNDIE), ANLIS, Ministerio de Salud de la Nación, Buenos Aires, Argentina; c Comité Nacional de Investigaciones Científicas y Técnicas (CONICET), Ministerio de Ciencia, Tecnología e Innovación Productiva, Argentina; d Instituto de Medicina Regional, Universidad Nacional del Nordeste, Resistencia, Chaco, Argentina. 122 Rado D1, Pérez JE4, Vilela M3, Quispe D1, Quispe A1, Aguilar EG1, Rangel E3, Valladares B2, Pacheco R1 1 Instituto Universitario de Enfermedades Tropicales y Biomedicina del Cusco – UNSAAC, Perú; 2Instituto Universitario de Enfermedades Tropicales y Salud Publica de Canarias, Universidad de La Laguna, Tenerife, España; 3Laboratório de Transmissores de Leishmanioses – Instituto Oswaldo Cruz/FIOCRUZ, Brasil; 4 Instituto de Medicina Tropical Alexander von Humboldt, UPCH, Lima, Perú [email protected] [email protected] Keywords: Peru, Sandflies, Cusco Keywords: vector population, Cutaneous Leishmaniasis, wing shape, wing size The province La Convención (Cusco, Perú) is an endemic area of Cutaneous Leishmaniosis, having the highest prevalence of the Department of Cusco. Aguas Calientes at 890 m. (Kepashiato Health Center) reports the highest frequency of cases. The objective of this study was to determine the sand fly fauna and its populations in Aguas Calientes. Sandflies were collected using Shannon traps in extradomiciliary environments and CDC light traps in intra, peri and extradomiciliary environments. The collections were done during dry and rainy seasons in 2010-2011. Thirty one species of Lutzomyia and Brumptomyia genus were identified with 1 new record for Peru: Lutzomyia sericea, 3 new record for Cusco: L. b. barrettoi, L. flaviscutellata, L. gantieri; 4 new records for La Convención: L. octavioi, L. migonei, L. ayrozai, L. guderiani and 2 not described species: L. (Psychodopygus) sp. (ca. L. (Psychodopygus) leonidasdeanei) and L. (Helcocyrtomyia) sp. (ca. L. (Helcocyrtomyia) kirigetiensis). The most predominant species were L. y. yuilli (47,25%) and L. octavioi (25,97%) with activity peaks during 21:00-23:00 and 4:00-06:00, respectively. This species were found in all collecting sites with predominace of L. y. yuilli in extradomiciliary environments. Nyssomyia neivai is the principal vector of Leishmania sp. causing epidemic Cutaneous Leishmaniasis in nine endemic provinces of Argentina. At the regional scale, studying the heterogenity of populations can give clues to develop common or differential control strategies, to study risk potentiality and migratory routes. If phenotypic variation has environmental and/or genetic causes, its study could help to detect local populations with potentially important characters. Geometric morphometry is a relevant tool for quantifying phenotypic variation and to understand its epidemiological importance. To assess differences in shape and size between Ny. neivai populations, we compared wing shape and size between individuals from three endemic northeastern provinces: Chaco (Puerto Antequeras, 31 females), Formosa (Las Lomitas, 19 females), and Misiones (Iguazú National Park, 16 females). We used 12 landmarks on each wing (image digitalizing: .TPSdig2, Procrustes method and relative warps calculation: TPSrelw).Centroid size was analized with a linear mixed model to allow for heterocedastic group variance, followed by Tukey contrasts (package lme, R software). Mean centroid size of females from Misiones was found to be greater from the ones from Formosa (mean (SE): 2285 (21); 2222 (12); p= 0.025). No evidence of centroid size differences were found between groups from Misiones and Chaco (p= 0.44), or Chaco and Formosa (p= 0.60).Wing shape was analized by MANOVA and HotellingBonferroni contrasts (Infostat). We found significant differences in wing shape between Misiones females and Chaco/Formosa groups (Wilks= 0.41; d.f.: 24, 104; p= 0.0012). Following a discriminant analysis (Axis 1= 66%), reclassification based on shape showed 71% of correct validated classification. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 124 ID 129-P Ecology & Epidemiology ID 130-P Ecology & Epidemiology STUDIES ON FAUNA OF PHLEBOTOMINE SAND FLIES (DIPTERA: PSYCHODIDAE) AT “TERRA FIRME” AND “VÁRZEA” ENVIRONMENTS IN TEFÉ MUNICIPALITY, AMAZONAS STATE, BRAZIL PHLEBOTOMINE SAND FLY SURVEY IN THE FOCUS OF HUMAN LEISHMANIASIS IN SOUTH WESTERN MADRID REGION, SPAIN: 2012-2013 Maribel Jiménez, Estela González, Sonia Hernández, Ricardo Molina Antonio Marques Pereira Júnior1,2, Eric Fabricio Marialva2, Moreno de Souza Rodrigues3, Felipe Arley Costa Pessoa3, Jansen Fernandes de Medeiros4 1 Instituto Nacional de Pesquisas da Amazônia, 2Centro de Pesquisa Leônidas & Maria Deane, FIOCRUZ AMAZÔNIA, 3Centro de Pesquisa Gonçalo Muniz, FIOCRUZ BAHIA, 4FIOCRUZ Noroeste [email protected] Unidad de Entomología Médica, Servicio de Parasitología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Ctra. Majadahonda-Pozuelo s/n, 28220, Majadahonda, Madrid, Spain. [email protected] Keywords: Phlebotomus perniciosus, Leishmania infantum, sand fly infections, Spain Keywords: Ecology, Amazônia, Epidemiology Phlebotomine sandflies are insects of medical importance because some female species are vectors of leishmaniasis. The aim of this work was to verify phlebotomine fauna at “Terra firme” and “Várzea” environments and ecotopes (forest and peridomiciles) in Tefé municipality, Amazonas state, Brazil. The collections were performed using HP light traps during six months: January, February, April, August, September and October, of 2013. The specimens were clarified with Potassium Hydroxide (10%) and mounted on slides. A total of 5.716 individuals were collected and identified 46 species. The abundance of the males was slightly higher compared to females, 2.868 males (50.19%) and 2.848 females (49, 81%), respectively. Trichophoromyia ubiquitalis (3,330 – 58.26%) and Nyssomyia antunesi (661 – 11.26%) were most abundant. At “Terra firme” environment 42 species were found (5.428 individuals) and Th. ubiquitalis, Ny. antunesi and Ny. yuilli yuilli were most abundant species with 3.312, 526 and 259 individuals, respectively. While at “Várzea” environment 23 species were found (288 individuals) and Ny. antunesi, Ev. walkeri were most abundant species with 135 and 87 specimens, respectively. The forest ecotopes had high diversity when compared to peridomiciles in both environments. At terra firme, the ecotope forest was richer with 4.689 specimens belonging to 39 species, while in peridomicile were found 739 individuals into 27 species. At várzea, the forest also most abundant with 263 individuals belonging to 21 species, while peridomiciles ecotope were found 25 specimens into 4 species. Since 2010 it has increased the number of cases of both visceral and cutaneous human leishmaniasis in south western Madrid region, Spain. Surprisingly the prevalence of canine leishmaniasis in the area of the focus is even lower than that detected in the neighbouring areas. In a preliminary entomological survey carried out in the area in 2011 Phlebotomus perniciosus was the only one potential vector identified. In order to a better understanding of the new active focus an entomological survey was carried out from May to October 2012 and 2013 in 4 stations located in a neighbouring green park next to the urban areas and selected according the higher number of sand flies collections obtained in 2011 in the same area. Each month 20 sticky traps (20 x 20 cm) and 2 CDC light traps were used during two consecutive nights in every station. CDC traps were replaced every day. The main objective was to establish the phenology of P. perniciosus in the focus and its relationship to the rates of infection of this species. A total of 29,729 specimens were collected during both years, 12,271 sand flies by CDC light traps and 17,458 using sticky traps. Three sand fly species were identified in 2012: P. perniciosus (68.57%), Phlebotomus sergenti (0.02%), and Sergentomyia minuta (31.41%). Data were similar in 2013 with the following species identified: P. perniciosus (64.35%), Phlebotomus papatasi (0.01%), and S. minuta (35.64%). Mean densities of P. perniciosus obtained using sticky traps were 159.01 flies/m2 in 2012 and 146.13 flies/m2 in 2013. On the other hand, 735 P. perniciosus female collected with CDC traps in the seasonal study of 2012 were dissected and 18 of them were found infected with L. infantum (2.45%). Infected females were found in the four stations from July to September. A total of 14 isolates were successfully isolated and further characterized as L. infantum by PCR of ITS regions. In 2013 a number of 863 P. perniciosus females were dissected with 57 of them infected, giving a rate of 6.6%. A total of 44 isolates were obtained in the 4 stations from July to October and were also characterized as L. infantum. We present here the evolution of L. infantum infection rates in P. perniciosus along the transmission season in the four stations studied during 2012 - 2013. These data provide significant epidemiological information which is closely linked to the spread of human leishmaniasis in the focus. This study was funded by EU grant FP72011-261504 EDENext. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 127-P Ecology & Epidemiology ID 128-P Ecology & Epidemiology LUTZOMYIA SPP. (DIPTERA: PSYCHODIDAE) IN INTRA, PERI AND EXTRADOMICILIAR ENVIRONMENTS IN YOMENTONI (RIGHT MARGIN), LA CONVENCIÓN, CUSCO, PERÚ SOME ECOLOGICAL ASPECTS OF LUTZOMYIA LONGIPALPIS (DIPTERA: PSYCHODIDAE) IN ENDEMIC AREA OF VISCERAL LEISHMANIASIS, MATO GROSSO DO SUL, BRAZIL. PRELIMINARY RESULTS Toccas F1, Chacon M1, Pérez JE3, Rado D1, Soto M1, Mendoza J1, Luna R1, Aguilar EG1, Valladares B2, Pacheco R1 1 Instituto Universitario de Enfermedades Tropicales y Biomedicina del Cusco – UNSAAC, Perú; 2Instituto Universitario de Enfermedades Tropicales y Salud Publica de Canarias, Universidad de La Laguna, Tenerife, España; 3Instituto de Medicina Tropical Alexander von Humboldt, UPCH, Lima, Perú [email protected] Keywords: Perú, Cusco, Lutzomyia Yomentoni at 583 m. (Right Margin) is an endemic area for leishmaniosis, no studies on Lutzomyia, spp. were done in the past. The objective of this study was to identify and determine the species of Lutzomyia in intra, peri and extradomestic. Sand fly collections were done with CDC light traps and Shannon trap with protected human attractant during the rainy and dry seasons of 2010 and 2011. Twenty eight sandfly species were identified, with a new record for the Province La Convención: L. (P.) trispinosa. Extradomiciliar environment (44.03%) shown the largest sand fly collections.L. (Pressatia) sp. was the most frequent species for the three collection environments and for both the rainy and dry seasons. As well as there is significant difference (p = 0.000<0.05) for species presence in the three collection environments in relation with the seasons and sex; most female collections belongs to L. (Pressatia) sp. and L. (Trichophoromyia) sp. Supported by AECID, Spain. 123 Helen Rezende de Figueiredo1, Aline Etelvina Casaril1, Jucelei O. M. Infran2, Elisa Teruya Oshiro3, Adauto Rodrigues Mendes3, José Dilermando Andrade Filho5, Letícia Moraes Ribeiro2, Everton Falcão Oliveira6, Mirella Ferreira da Cunha Santos2, Alessandra Gutierrez de Oliveira1, 2, 3 1 Post Graduate Program in Infeccious and Parasitic Diseases, Federal University of Mato Grosso do Sul; 2Post Graduate Program in Animal Biology, Federal University of Mato Grosso do Sul; 3Parasitology Laboratory, Federal University of Mato Grosso do Sul; 4Central of Zoonotic Control, Municipality of Aquidauana/MS; 5Center of National and International Reference in Sandflies, Leishmaniasis Laboratory, Research Center René Rachou; 6Post Graduate Program in Public Health, São Paulo University. [email protected] Keywords: Vector, sandfly, seasonality, behaviour The municipality of Aquidauana, MatoGrosso do Sul, Brazil, frequently presents cases of canine and human visceral leishmaniasis and is classified as an area of intense transmission. The sandfly Lutzomyia longipalpis is the major vector of Leishmania (Leishmania) infantum, the causative agent of visceral leishmaniasis in Central and South America. This vector is widely distributed in both wild and domestic surroundings. This species occurs in areas with different climatic conditions and presents great adaptability to different habitats. This study aimed to identify behavioral aspects of Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae) in the study area The captures were carried out from April 2012 to March 2014, in six neighborhoods, fortnightly, with light automatic traps from 06 pm to 07 am, in peri and intradomicile environments each dwelling. The specimens were transported to the Laboratory of Parasitology of the Federal University of MatoGrosso do Sul (UFMS) for mounting, identification, and other observations. During the capture period 2569 specimens of Lutzomyia longipalpis, 2271 males (88.4%) and 298 females (11.6%) were collected. Of the total, 71.7% (1842) were captured in the first year, which showed irregular distribution over the months. In the second year, the distribution of the remaining 29.3% (727 individuals) seems to be more regular, coinciding peaks of higher frequency in wet seasons and peaks of less frequency in dry seasons. Lu.longipalpis behaved as an annual species, but was not present in all locations through the month. The population of the neighborhood 2 (Pinheiro) had the highest percentage of sand flies (31.9% - 820 individuals), despite not having been captured in all months. On the other hand, the population of the neighbourhood 4 (São Cristóvam) had the second highest percentage of Lutzomyia. longipalpis (28.6% - 737 individuals) and was present in all months. The sandflies were captured more frequently peridomiciles 81.08% (2,083) than in households 18.91% (486), important information, considering the anthropophilic behavior of this vector. In addition, females were analyzed by polymerase chain reaction (PCR) in search of Leishmania sp. Analysis was negative, which was expected, since the natural infection rates, even in endemic areas, tend to be very low. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 131-P Ecology & Epidemiology ID 132-P Ecology & Epidemiology MOLECULAR DETECTION OF LEISHMANIA INFANTUM AND HOST-FEEDING PREFERENCES IN PHLEBOTOMUS PERNICIOSUS FROM THE FOCUS OF LEISHMANIASIS IN SOUTH WESTERN MADRID REGION, SPAIN: 2012-2013 INTERACTION BETWEEN CANINE AND HUMAN VISCERAL LEISHMANIASES IN A HOLOENDEMIC FOCUS OF CENTRAL TUNISIA Maribel Jiménez, Estela González, Sonia Hernández, Ricardo Molina Unidad de Entomología Médica, Servicio de Parasitología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Ctra. Majadahonda-Pozuelo s/n, 28220, Majadahonda, Madrid, Spain. [email protected] Keywords: Phlebotomus perniciosus, host feeding preferences, Leishmania infantum, Spain. An entomological survey was carried out from May to October 2012 and 2013 in 4 stations placed neighbouring the urban area of the focus of leishmaniasis in south western Madrid. Direct xenodiagnosis in hares (Lepus granatensis) and wild rabbits (Oryctolagus cuniculus) from the focus proved that they are infective to colonized Phlebotomus perniciosus. Further characterization of promastigotes isolated from these sand flies fed on both hares and rabbits demonstrated that these lagomorphs were infected by Leishmania infantum. The analysis of blood preferences and detection of L. infantum was studied in a total of 139 blood female sand flies captured in the transmission period from June to October of 2012 (n=71) and 2013 (n=68). CDC light traps and sticky traps were used placed in four stations through the entomological surveys. Initially, genomic DNA was obtained from individual sand flies and used to amplify a 120 bp fragment of the conserved region of kDNA.. Positive samples were further analysed following a specific L. infantum PCR based on the amplification of a fragment of 702 bp of cpb gene. Blood meal identification was conducted by the amplification of a fragment of 359 bp of vertebrate cytochrome b (cyt b) gene, further sequencing, and subsequent comparison with sequences deposited in the GenBank. Data from female sand flies captured in 2012 revealed that blood meals were mainly rabbits (n=41; 57.8%) followed by hares (n=28; 39.4%). DNA from L. infantum was detected in (n=2; 4.9%) sand flies fed on rabbits and in (n=11; 39.3%) sand flies fed on hares. Similarly, blood source from a human and another one from a dog were respectively found in two sand flies representing 1.43% in each case. In addition, in the 68 sand flies processed during 2013 blood meal from rabbits mostly was found in 46 specimens (67.6%), and in 16 (23.5%) blood source from hares was identified. DNA from L. infantum was detected in ten (21.7%) sand flies fed on rabbits and in three (18.8%) fed on hares. Also, in five sand flies blood from cats was found (7.3%) and in one sand fly (1.5%) human blood was identified. In conclusion, the data revealed an important increase in the number of sand flies fed on rabbits during the sand fly season, a direct consequence of the control measures undertaken all over 2012 that significantly reduced the number of hares in the green park close to the urban area of the outbreak.This study was funded by EU grant FP72011-261504 EDENext. 125 Z Zoghlami1, E Chouihi1, W Barhoumi1, K Dachraoui1, Nabil Massoudi1, K Ben Helel2, Z Habboul2, MH Hadhri§, S Limam3, M Mhadhbi3, M Gharbi3, E Zhioua1 1 Institut Pasteur de Tunis, Laboratory of Vector Ecology, 13 Place Pasteur BP 74, 1002 Tunis, Tunisia. 2Paediatric Department, University Hospital of Kairouan, Tunisia, §.Regional Department of Health, governorate of Kairouan, Tunisia. 3Veterinary School of Sidi Thabet, Sidi Thabet, Tunisia [email protected] Keywords: Visceral leishmaniasis, Dogs, Humans, Sand fly vectors Canine visceral leishmaniasis (CanVL) is endemic in the Mediterranean basin. In Tunisia, CanVL is spatially associated with human visceral leishmaniasis (HVL) affecting mostly children younger than 5 years old. In this study, seroprevalence of Leishmania infantum infection in dogs was assessed in highly endemic districts of the governorate of Kairouan where more than 50% of HVL cases in Tunisia were reported. An entomological investigation was also carried out in two endemic districts (Bouhajla and Haffouz) to assess sand fly fauna and infection status of sand flies with Leishmania. A total of 191 serum samples were collected from healthy dogs and tested for anti-L. infantum antibodies by indirect immunofluorescence antibody test (IFAT). Overall seroprevalence was 26.7% being highest among dogs in the district of Bouhajla (52.7%) and the lowest in the district of Chbika (5.2%). In dogs, seroprevalence did not differ significantly based on gender or age, with dogs younger than 1 year showing a higher seroprevalence compared to older dogs. These findings suggest strong force of infection in naïve animals in holoendemic regions leading to emerging high incidence of HVL. Concomitant to the high CanVL prevalence observed in the Bouhajla district, a significantly high cumulative HVL incidence also was observed in this district. Phlebotomus perniciosus and Phlebotomus longicuspis were the most abundant sand fly species in Bouhajla and Haffouz districts. The rate of Leishmania-DNA infection in sand flies was 9.4%. This finding points to spatial correlation between the occurrence of disease in humans, a high rate of infection in dogs and a high abundance of P. pernicious and P. longicuspis. Thus, CanVL is the main risk factor for transmission to humans and subsequently, it is an important parameter for controlling transmission to humans. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 133-P Ecology & Epidemiology ID 134-P Ecology & Epidemiology LEISHMANIASIS VECTORS IN THE ALTO PARANÁ DEPARTMENT, PARAGUAY STUDIES ON SAND FLIES (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) OF URBAN AND WILD AREAS IN THE DISTRICT OF BARRA DO GUAICUI, VÁRZEA DA PALMA MUNICIPALITY, MINAS GERAIS, BRAZIL Nilsa Gonzalez-Britez1, Nilda Portillo1, Maria Ferreira 1, Martha Torales2, Nidia Martínez3, Luciano Franco3 126 1 .Dpto. de Medicina Tropical. Instituto de Investigaciones en Ciencias de la Salud, Universidad Nacional de Asunción. 2 Dpto. de Entomología del Servicio Nacional de Erradicación de Paludismo SENEPA. 3 Dpto. Programas del SENEPA-Ministerio de Salud Publica y Bienestar Social Danyele Franca da Silva1, Cristiani de Castilho Sanguinette1, Gabriel Barbosa Tonelli1, Aline Tanure1, Rodolfo German Antonelli Vidal Stumpp2, Felipe Dutra Rêgo1, Célia Maria Ferreira Gontijo1, José Dilermando Andrade Filho1 [email protected] 1 2 Centro de Pesquisas René Rachou/FIOCRUZ- Belo Horizonte, Minas Gerais. Universidade Federal de Minas Gerais- Belo Horizonte, Minas Gerais Keywords: Leishmaniosis, Phlebotominae, Alto Paraná. [email protected] Leishmaniasis is a complex diseases caused by protozoa of the genus Leishmania and are transmitted by the bite of blood-sucking insects, known in Paraguay under the name of Karachá, belonging to several genera of the family Psychodidae, subfamily: Phlebotominae. This work was developed based on the entomological collections made in several districts of Alto Paraná, with the objective to identify the species present in the border region between Brazil, Paraguay and Argentina, and contribute with strategies based on entomological surveillance. Secondary data was used from the sand flies collected by the National Service Malaria Eradication using CDC light traps, containing dry ice, placed 1.5 meters above the ground for two nights per place. The traps were switched on at 19:00 hrs and the collections were made the following day at 9:00hrs. The study was carried out between the periods of June to November 2012. Were selected collection areas representing different environments favorable for the vector covering several localities of the Department of Alto Paraná. Were identified nine species of Plhebotominae, of which three are of epidemiological importance in the transmission of Leishmania braziliensis. In order of frequency vectors identified were: Nyssomyia neivai, collected in San Alberto, Mbaracayú, Hernandarias and Los Cedrales districts; Migonemyia migonei, in San Alberto, Mbaracayú and Los Cedrales districts; and finally Nyssomyia whitmani in San Alberto and Mbaracayú districts. The occurrence of vectors in districts including rural, semi-urban and urban areas of the border region, is considered a risk factor for the transmission of cutaneous leishmaniasis in these areas. Moreover, studies with sandflies are important to acquire adequate knowledge about the distribution areas of circulation and population dynamics of the vectors, which is an important tool for control strategies aimed at reducing the risk of contact with the vector, as well as the implementation of policies for the epidemiological control of leishmaniasis. Keywords: Leishmaniasis, Lutzomyia longipalpis, Nyssomyia intermedia : Leishmaniasis are chronic diseases caused by protozoa of the genus Leishmania. The main mode of transmission of the parasite to humans and other mammalian hosts occurs through the bite of some infected female sand flies. The municipality of Várzea da Palma is located in the north of Minas Gerais State, presenting 202 human cases of autochthonous cutaneous leishmaniasis (CL) and 37 cases of visceral leishmaniasis (VL) recorded between 2004 to 2013. Knowing that the spread of leishmaniasis depends mainly on the presence and distribution of vectors, the correct identification of the species of sand flies circulating in endemic areas and knowledge of their ecology can contribute to the knowledge of leishmaniasis. This study aims to identify and compare the sand fly fauna in different environments. The study was conducted in the district of Barra do Guaicui, belonging to the municipality of Várzea da Palma. From March 2013 to February 2014, monthly, two CDC light traps were installed in the transition area, six in wild area and two traps in the urban area. A total of 3,445 sand flies belonging to eight genera and 15 species were captured. Nyssomyia intermedia (35.99%), Lutzomyia longipalpis (17.68%), Evandromyia lenti (14.98%), Evandromyia evandroi (11.12%) were the species with the highest population density. The urban area accounted for 60.46% of total sand flies collected, followed by wild area (32.80%) and the transition area (6.73%). Nyssomyia intermedia, Lutzomyia longipalpis and Nyssomyia neivai were more abundant in the urban area. Evandromyia lenti and Evandromyia evandroi had high density in the wild area. The most representative species in the transition area was Brumptomyia avellari. Evandromyia sallesi was evenly distributed between urban and wild areas. Important vector species involved in the transmission of CT and VL in the area, some of which may have a role in the urban and wild cycle of leishmaniasis in Barra do Gauicuí area. Financial Support: PAPES/FIOCRUZ, CNPq, CAPES and FAPEMIG N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 135-P Ecology & Epidemiology ID 136-P Ecology & Epidemiology E VA L U AT I O N O F T H E A B U N D A N C E O F PHLEBOTOMINE SAND FLIES IN HOUSEHOLDS FROM AN AREA OF VISCERAL LEISHMANIASIS TRANSMISSION IN BELO HORIZONTE, MINAS GERAIS, BRAZIL PHLEBOTOMINAE SAND FLY FAUNA IN TWO URBAN PARKS IN BELO HORIZONTE, MINAS GERAIS, BRAZIL (DIPTERA: PSYCHODIDAE) Elisa Neves Vianna1, Maria Helena Franco Morais2, Andréa Sobral de Almeida3, Paulo Chagastelles Sabroza3, Mariângela Carneiro4 & Edelberto Santos Dias5 Faculdade de Medicina, Universidade federal de Minas Gerais. [email protected] Keywords: receptivity, vulnerability, phlebotomines Since 1994, Visceral Leishmaniasis (VL) has been suffering a process of urbanization in Belo Horizonte, with an increase in both canine and human cases throughout the years. The vector species, Lutzomyia longipalpis, is present in the entire municipality, both in the peri and intradomicile. The identification of receptive (presence of organic material, trees, shady spots, available hosts) and vulnerable households (presence of attics in the domicile, cracks, broken windows) with the presence of phlebotomines could aid the actions of the Ministry of Health. The objective of this study was to validate the classification of receptive/vulnerable households for the colonization by phlebotomine sand flies and the risk of VL transmission through an entomological survey. One-hundred and fifty-three properties located in neighborhoods from the Northeast Region of Belo Horizonte were classified into three levels of receptivity/vulnerability (low, medium and high) by the endemic combat agents. A random sample of 46 of the 153 properties included in the study was used to obtain data for the entomological survey. These properties were prospected bimonthly between May 2012 and June 2013 for three consecutive nights, using HP traps, one in the peridomicile and another within the domicile. Among the households classified as high and medium receptivity/vulnerability, 86% (10) and 77% (25) of them, respectively, presented infestation by phlebotomines in at least one of the bimonthly evaluations. The classification levels of the households did not present an association with the abundance of phlebotomines as measured through Spearman´s correlation (ρ=-0,7, p >0,05). In total, 752 phlebotomines were captured, 621 in the peridomicile and 131 within the domicile, showing greater captures in January and March. Although there was no significant correlation among the different household classification levels and the abundance of phlebotomines, the households presented an appreciable percentage of infested houses, making them productive for the insects. The greatest number of phlebotomines was found in the peridomicile, probably due to the greater availability of food sources in the environment. The species of phlebotomine sand flies are being identified and may be related to environmental and climatic variables, highlighting issues of the ecoepidemiology of VL in the region and aiding future entomological survey actions for the Visceral Leishmaniasis Control Program. 127 Paula Cavalcante Lamy Serra e Meira, Bruna Lacerda Abreu, Ana Paula Lusardo de Almeida Zenóbio, Cristiani de Castilho Sanguinette, Gustavo Mayr de Lima Carvalho, Lara Saraiva, José Dilermando Andrade Filho Centro de Pesquisas René Rachou/FIOCRUZ [email protected] Keywords: Phlebotominae, Leishmania, cave Phlebotomine sand flies are often collected in urban areas in Brazil. This fact is very important, since some species may be involved in the transmission of Leishmania to mammals. Based on this background, this study aimed to determine the sand fly fauna in the cave environment in the Parque Municipal das Mangabeiras and Parque Paredão Serra do Curral, located in the urban area of Belo Horizonte, Minas Gerais, Brazil. In order to study the sand fly fauna, systematized collections were conducted for a year, with monthly captures from November 2011 to October 2012, using automatic CDC light traps and non-systematized collections using Shannon trap. Thus, the sand fly fauna of Parque Municipal das Mangabeiras presented fourteen species, being the most abundant Evandromyia edwardsi and Parque Paredão Serra do Curral featured four species, being the most abundant Sciopemyia microps. In both parks were collected species that transmit leishmaniasis, such as Lutzomyia longipalpis, Nyssomyia intermedia and Nyssomyia neivai. Studies on the fauna and the behavior of sand flies in cave environments are essential to understand the relationship of these insects with this ecotope and also assess and monitor areas that may pose risks to the health of visitors and employees in this type of environment.Financial Support: FAPEMIG, CNPq. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 137-P Ecology & Epidemiology ID 138-P Ecology & Epidemiology SPECIES OF PHLEBOTOMINE SAND FLIES (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) REPORTED IN BRAZILIAN CAVES ECOLOGY OF SAND FLIES (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) IN THE NATURAL HERITAGE PRIVATE RESERVE OF SERRA DO CARAÇA, MINAS GERAIS, BRAZIL Paula Cavalcante Lamy Serra e Meira and José Dilermando Andrade Filho Centro de Pesquisas René Rachou/FIOCRUZ [email protected] 128 Gabriel Barbosa Tonelli, Felipe Dutra Rego, Gustavo Mayr de Lima Carvalho, Aline Tanure, José Dilermando Andrade Filho Centro de Pesquisas René Rachou/FioCruz - Belo Horizonte, Minas Gerais [email protected] Keywords: Phlebotominae, cave, Evandromyia Keywords: Ecotourism; Phlebotominae; Leishmaniasis; Transmission. Among the insects that are found in the cave environment, sand flies are noteworthy, since they are the main vectors of some pathogens of medical and veterinary importance, among which stand out Leishmania. Many caves, even before being studied, are open for visitation and work practices, without any scientific monitoring. Thus, the loss of geological, biological and ecological information is immense. In addition, visits to areas where fauna surveys have not been conducted, especially of the arthropods presents in the locations, can pose risks to the health of the population who visit and work there. Based on this background, this study aimed to report the species of sand flies collected in Brazilian caves. A literature review was conducted and the species of sand flies, the lithology of these caves and the state where the registration took place. Until now, 84 species of phlebotomine belong to 17 genus were reported in Brazilian cave, of these species, 14 were described from caves. The Evandromyia genus is the most representative in Brazilian caves with 15 species, followed by Lutzomyia and Micropygomyia, both with 11 species caves. Thus, only the species of these three genera, represent 44.05% of sand flies from caves. Seventeen species reported in Brazilian caves can be involved in the transmission of Leishmania to mammals. Given the information above, it is clear that the Brazilian caves are an important place to collect sand flies, since several species were described from caves and this ecotypes can host important species are important in the transmission of Leishmania. Financial support: Fapemig, CNPq There are several tropical vector-borne diseases and among them are leishmaniases. This complex parasitic disease caused by protozoa, include approximately 21 species of Leishmania, several mammalian and sand flies species, which act as reservoirs and vectors of Leishmania. Knowledge of its transmission cycle can provide understanding of the epidemiology and ecological relationships of leishmaniasis. The ecotourism is a growing activity in Brazil. The increase in tourism in the country associated with the medical importance of sand flies, justify more detailed studies to know the possible health risks of tourists and workers in these locations. The Natural Heritage Private Reserve Serra do Caraça (RPPNPC) is located in the municipalities of Santa Barbara and Catas Altas and covers an area of 11,233 hectares, with transitional vegetation between Cerrado and Atlantic Forest. The aim of this study was to study the sand fly fauna of this site and verify the presence of possible vector species of Leishmania, to contribute to the control of leishmaniasis among tourists in RPPNPC. We used 25 CDC light traps distributed in seven trails in the park. Trails 1 and 2 were exposed in forested areas; trials 3 and 4 in “campo rupestre” and in one cave; trials 5 and 6 in the peridomestic areas; and trial 7 in the intradomicile (in the house provided for researchers) and outer wall of the house. Collections are being carried out every two months for one year. To date six samplings were performed, with 342 sand flies collected, distributed in 9 genera and 17 species. The predominant species were Psychodopygus lloydi (71.4%) and Pintomyia monticola (6.14%) and the places where there was a greater number of sand flies were the trail 7 (27.19%), 1 and 6 (24.27%). In recent studies, specimens of Ps. lloydi were found infected with Leishmania braziliensis in Minas Gerais, which may be an indication that this species may be involved in the transmission of Leishmania in this area. Financial support: FAPEMIG e CNPq. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 139-P Ecology & Epidemiology ID 140-P Ecology & Epidemiology STUDY OF SAND FLIES (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) IN A TRANSMISSION AREA OF LEISHMANIASIS IN MINAS GERAIS, BRAZIL ASSESSMENT OF SAND FLIES (DIPTERA: PSYCHODIDAE) IN AN AREA WITH CANINE VISCERAL LEISHMANIASIS IN THE MUNICIPALITY OF PORTO ALEGRE, RIO GRANDE DO SUL, BRAZIL Aline Tanure, Gabriel Barbosa Tonelli, Felipe Dutra Rêgo, Gustavo Mayr de Lima Carvalho, Célia Maria Ferreira Gontijo, Gustavo Fontes Paz, José Dilermando Andrade Filho Centro de Pesquisas René Rachou/FioCruz- Belo Horizonte, Minas Gerais [email protected] Keywords: Phlebotominae, Leishmaniasis The intense process of urbanization and industrialization cause a series of changes in urban areas, extending big cities and creating metropolitan regions. This is the case of the metropolitan region of Belo Horizonte, capital of Minas Gerais State, which consists of 16 municipalities, including Brumadinho. In recent years Brumadinho has undergone considerable socio-environmental changes. Casa Branca is a district of Brumadinho which is undergoing intense transformation process, where the construction of condominiums attracts people in search of tranquility and quality of life. However, it is known that changes in the natural environment are important factors in the emergence and reemergence of infectious and parasitic diseases, including leishmaniasis. The municipality of Brumadinho have reported an increase in cases of cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL) in the last 5 years, the latter classifying the program as an area of sporadic transmission, according to the Ministry of Health, but, transmission of the disease has not yet been elucidated since no studies on the sand fly fauna are available. This project investigates the sand fly fauna in Casa Branca area. For this, 18 CDC light traps were used for three consecutive nights, exposed in the peridomicile of 9 homes. Until now, a total of 5,284 sand flies were collected belonging to 13 species, Nyssomyia whitmani (88.9%) were the most abundant species, followed by Lutzomyia longipalpis (5.86%). Both species are involved in the transmission of CL and VL which reinforces the need to investigate the sand fly behavior aspects in Casa Branca and understand their role in the chain of transmission of leishmaniasis and direct preventive and control measures. Financial Support: FAPEMIG, CNPq. 129 G D Souza1, 2; A Cardoso1; R M J S de Carvalho1 1 Núcleo de Vigilância de Roedores e Vetores, Coordenadoria Geral de Vigilância em Saúde, Secretaria Municipal de Saúde de Porto Alegre, RS, Brazil; 2Seção de Reservatórios e Vetores, Divisão de Biologia Médica, Instituto de Pesquisas Biológicas-Laboratório Central de Saúde Pública, Fundação Estadual de Produção e Pesquisa em Saúde, Secretaria Estadual de Saúde do Rio Grande do Sul, RS, Brazil. [email protected] Keywords: Canine visceral leishmaniasis; sand flies; Porto Alegre; Brasil Canine visceral leishmaniasis (CanVL) was confirmed in the state of Rio Grande do Sul, Brazil, in 2008. Two years later, the disease was diagnosed in dogs in a periurban area, in the city of Porto Alegre, Rio Grande do Sul. Thus, the objectives of this study were to survey the sand flies species present in this region, to determine the epidemiological important of these species in the transmission of this disease and to understand the population dynamics of these vectors to help vector control activities. Sand flies were collected monthly from February/2012 to January/2013 in the locality of Lageado, with CDC light traps, in three environments: inside house, in the peridomestic area (domestic animal shelter) and in the forest. The sampling effort was to capture two nights per month, with one trap per environment. A total of 3,397 sand flies were collected in the three studied environments, consisting of 1,575 males and 1,822 females. The species identified were Migonemyia migonei (62.88%), Pintomyia fischeri (22.49%), Nyssomyia neivai (11.89%), Psathyromyia lanei (1.77%), Brumptomyia sp. (0.77%), Pintomyia monticola (0.18%) and Lutzomyia gaminarai (0.03%). Migonemyia migonei showed the highest relative frequency of occurrence, representing 58.2% of specimens captured inside house, 65.64% in the peridomestic area and 61.61 % in the forest, followed by Pi. fischeri (22.58%, 23.66% and 19.27%, respectively) and Ny. neivai (16.63%, 9.06% and 13.28%, respectively). The largest number of sand flies (n=1,822) was collected in the peridomicile and in the forest the highest diversity was found: seven species. Regarding the annual distribution, almost all sand flies (95.14%) were collected from November to May, with a large peak of specimens collected in February. The three main species were found in all months of the year, except for Pi. fischeri in July. The predominance of Mg. migonei suggests this species as the main suspect to transmit visceral leishmaniasis to dogs in the region. Pi. fischeri and Ny. neivai are known important vectors in the transmission of cutaneous leishmaniasis in Porto Alegre and they can also act as secondary vectors for CanVL. For prevention and control, environmental management should be intensified on the spring and summer. If necessary, chemical control should be realized in October, with reapplication of insecticide in the following January. Supported by: Municipal government of Porto Alegre. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 141-P Ecology & Epidemiology ID 142-P Ecology & Epidemiology PERSISTENCE OF LUTZOMYIA LONGIPALPIS, VECTOR OF VISCERAL LEISHMANIASIS, IN THE CITY OF SALTO, URUGUAY SEROLOGICAL SURVEILLANCE IN DOGS OF THE CITY OF BELLA UNIÓN, A RISK AREA FOR VISCERAL LEISHMANIASIS Yester Basmadjián1, Bruno Canneva1, Lorenzo Verger2, Alejandra 2 1 Lozano , Dinora Satragno , Eduardo Supparo2,3, Cirino Sequeira4, Selva Romero1, Ana Viera1, Cecilia Tort1, Cristina Ríos 5, Patricia Lagarmilla5 and Edgardo Vitale 2 Verger L1; Lozano A1; Vitale E1; Satragno D2; Sequeira C3; Canneva B2; Basmadjián Y2; Rios C4; Lagarmilla P4; Tort C2; Viera A2; Romero S2 1 Departamento de Parasitología y Micología Médica. Instituto de Higiene-Facultad de Medicina-UdelaR. Montevideo. Uruguay. Avda. Alfredo Navarro 3051. [email protected]; 2Área de Medicina Preventiva y Epidemiología, Facultad de Veterinaria, UdelaR. Montevideo. Uruguay; 3Liberal Veterinary Professional, Bella Unión, Departamento de Artigas, Uruguay; 4Área de Salud Pública Veterinaria, Facultad de Veterinaria, UdelaR. Montevideo. Uruguay5Área de Salud Pública Veterinaria, Facultad de Veterinaria, UdelaR. Montevideo. Uruguay. 130 1 Área de Medicina Preventiva y Epidemiología, Facultad de Veterinaria, Universidad de la República, Montevideo. Uruguay. Alberto Las places 1550. 2Departamento de Parasitología y Micología Médica. Instituto de Higiene, Facultad de Medicina, Universidad de la República. Montevideo. Uruguay. 3Liberal Veterinary Professional, Bella Unión, Departamento de Artigas, Uruguay. 4Área de Salud Pública Veterinaria, Facultad de Veterinaria, Universidad de la República. Montevideo. Uruguay. [email protected] [email protected] Keywords: Uruguay, dogs, Leishmaniasis Keywords: Lutzomyia longipalpis, Uruguay, Salto The city of Bella Union is located in the department of Artigas, in the northern part of Uruguay. In the year 2010, in this city, two specimens of L. longipalpis were found for the first time in the country. Further research has shown several months of continuous presence of the vector since the year 2013. These findings confirm the city as a risk area for an outbreak of Visceral Leishmaniasis (VL), an exotic disease in Uruguay. In the present work, we made a serological survey of canines using a rapid diagnostic kit for VL (DPP® Leishmaniose Visceral Canina, BioManguinhos). The sampling was aimed at dogs showing clinical signs of the disease (skin lesions, cachexy, fever) and at those who lived inside a 200 meters radius from a site where specimen of L. longipalpis were caught with a CDC light trap. 200 dogs where tested for VL: 170 living near sites infested with L. longipalpis and 30 with compatible symptoms. All the test results were negative. Despite not having been able to obtain a positive result to the test, further serological samplings are projected in order to keep the epidemiological surveillance of the risk areas. Leishmaniasis emerged in the Southern Cone in its human visceral form in the year 2000, in the Paraguay-Argentina border. Since then, the rapid expansion to the south of its vector, Lutzomyia longipalpis, has determined the appearance of human cases in Argentina in the year 2006 and the presence of infected dogs in the Argentinean city of Montecaseros, near the border with Uruguay, in the year 2009. With this background, in February of 2010, an investigation was conducted in order to determine the presence of the vector in two cities of the northern part of Uruguay; Salto (capital city of the department with the same name) and Bella Unión (department of Artigas), catching specimen of L. longipalpis in both cities. These findings determined that both departments were classified as “vulnerable” to the emergence of Visceral Leishmaniasis. After this punctual vector surveillance, no further research was made. In the present work an entomologic survey was conducted to determine if the vector was still present in the city of Salto. In February of 2014, two CDC light traps were placed two nights in a row in the city Zoo, the same location that resulted positive in the year 2010 (S 31°23'51.19" W 57°57'51.65"). The insects caught in the traps were identified under stereomicroscope and the Phlebotominae sand flies were mounted with Canada balsam for species identification. Tree specimens of L. longipalpis were identified: two males and one female. These results confirm that the presence of the vector in the year 2010 was not an isolated event and rather suggest a continuous presence in the area. The status of the department as vulnerable for the entry and dissemination of the causative agent of Visceral Leishmaniasis is reafirmed. The city of Salto is still the southernmost location were L. longipalpis was found in Uruguay. This project was finnanced by the program I+D 2012 of the Comisión Sectorial de Investigación Científica, Universidad de la República. This project was finnanced by the program I+D 2012 of the Comisión Sectorial de Investigación Científica, Universidad de la República. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 145-P Ecology & Epidemiology ID 146-P Ecology & Epidemiology IDENTIFICATION OF BLOOD MEAL IN LEISHMANIA INFANTUM VECTORS FROM THE NORTHEAST OF SPAIN: PHLEBOTOMUS ARIASI AND P. PERNICIOSUS BY POLYMERASE CHAIN REACTION BASED AND SEQUENCING METHODS PHLEBOTOMINE SAND FLIES (Diptera: Psychodidae) IN A PERI-URBAN SETTING IN THE METROPOLITAN REGION OF NATAL BRAZIL Ballart C1, 2; González E3; Ravel C4; Jiménez M3; Abras A1, 2; Molina R3; Portús M1; Gállego M1, 2 131 Maria de Lima Alves, Maria de Fátima Freire de Melo Ximenes, Marcos Paulo Gomes Pinheiro, José Hilário Tavares da Silva, Tamy Elicia da Silva Lopes, Marcel Miranda de Medeiros Silva Universidade Federal do Rio Grande do Norte – UFRN -1 Laboratori de Parasitologia, Facultat de Farmàcia, Universitat de Barcelona (UB), Spain; 2Centre de Recerca en Salut Internacional de Barcelona (CRESIB), Hospital Clínic-UB, Spain; 3Unidad de Entomología Médica, Servicio de Parasitología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Spain; 4Université of Montpellier, UMR5290 MiVEGEC et Centre National de Référence des Leishmanioses, Montpellier, France [email protected] Keywords: Phlebotomine sand flies, climatic variables, peri-urban area [email protected] Keywords: Leishmaniosis, P. ariasi, P. pernciosus, blood meal In Spain, human leishmaniosis is present as a hypoendemic disease. The epidemiological cycle is considered to be zoonotic, with dogs acting as the main reservoir. However, other mammals have also been implicated as reservoirs for Leishmania infantum, the only autochthonous species in the country. Two sand fly species belonging to the genus Phlebotomus subgenus Larroussius are proven vectors, P. ariasi and P. perniciosus. In some foci the species do not overlap, whereas in other they are found in sympatric conditions sharing the vectorial role. While awareness of the disease dates back to the beginning of the 20th century, it has little history in northern Spain, where new endemic areas have been found. Since the vectorial transmission of Leishmania depends on sand fly feeding behavior, the analysis of their blood meals is of epidemiological interest. Sand flies where captured with light traps (CDC-like) in northeast Spain (provinces of Lleida and Girona, Catalonia). Traps were set for one night in 20 localities near farms or animal stables and kennels. In total, 165 blood-fed sand fly females were analyzed: 121 P. ariasi, 41 P. perniciosus, and 3 unidentified. The assays were based on specific amplification and sequencing of the blood meal–derived single copy prepronociceptin (PNOC) gene or the vertebrate cytochrome b (cyt b) gene. Mixed blood meals were analyzed by PCR-RFLP. A Genbank search was carried out for sequence homology. Blood meals were identified in 39 flies, 25 P. ariasi, 11 P. perniciosus and 3 unidentified. Among these, blood meals from the following species were identified: Homo sapiens (3), Canis familiaris (2), Bos taurus (10), Ovis aries (15), Capra hircus (3), Equs caballus (3), Sus scofra (3), Oryctolagus cuniculus (1), Mus musculus (1), Gallus gallus (1). Mixed meals were identified in 4 cases (1 H. sapiens/O. aries, 3 Capra hircus/Ovis aries). The results related to the sand fly species and sample origin (location of traps and animals present) are discussed. Financial support: Spanish Ministerio de Ciencia e Innovación (CG12010-22368-C02-01). Phlebotomine sand flies are distributed in nearly all faunal regions worldwide as pathogenic vectors. In Brazil there are a large number of species. Northeast Brazil has the greatest occurrence of human cases, either separately or concomitantly of Visceral Leishmaniasis (VL) and American Cutaneous Leishmaniasis (ACL). The aim was to identify species of phlebotomine sand flies in a peri-urban area of the municipality of São Gonçalo do Amarante, Rio Grande do Norte state, where cases of ACL and VL have been notified. Captures were made monthly from January 2012 to December 2013 with CDC light traps, over 12 consecutive hours from 5:30 in the evening to 5:30 in the morning. A total of 272 specimens were captured in 2012 and 377 in 2013, 63% male and 37% female. Six species were identified: Evandromyia lenti 84%, Evandromyia evandroi 8%, Lutzomyia longipalpis 6%, Sciopemyia sordellii 1%, Evandromyia walkeri 0.5% and Nyssomyia intermedia 0.5%. The results show that Evandromyia lenti was more abundant, captured in every month of the year and in the three environments: shrubland 7.5%, intra-domicile 8.5% and peri-domicile 84%. Temperature and relative humidity apparently did not interfere in phlebotomine density in the study period. Greater species diversity occurred in 2012, with six species. Although 2012 and 2013 are considered drought years in the Northeast, a significant increase in Evandromyia lenti density was observed in March, April and May, coinciding with the low rainfall of this period. Considering the urban growth in the metropolitan region of this municipality, where human cases of visceral and cutaneous leishmaniasis have been recorded in recent years and E. lenti and L. longipalpis captured in the peri-domicile and intra-domicile, entomological and epidemiological surveillance are essential and must be associated to educational actions in public and environmental health in this municipality. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 143-P Ecology & Epidemiology ID 144-P Ecology & Epidemiology SANDFLIES (DIPTERA, PSYCHODIDAE, P H L E B O TO M I N A E ) I N P I TO C O C AV E , A N ARCHEOLOGICAL SITE IN NORTHERN MATO GROSSO DO SUL STATE, BRAZIL FIRST REPORT OF LUTZOMYIA LONGIPALPIS IN URUÇUCA, COCOA ZONE OF THE SOUTHERN STATE BAHIA, BRAZIL Eunice Aparecida Bianchi Galati1, Paulo Silva de Almeida2-3, Marcia Bicudo de Paula1, Pedro Catarino Costa Filho4, Romoaldo Martins de Almeida4 1 Universidade de São Paulo (USP), São Paulo, Brazil. 2Universidade Federal da Grande Dourados (UFGD), Mato Grosso do Sul, Brazil. 3Laboratório Regional de Entomologia, Dourados, Mato Grosso do Sul, Brazil. 4Coordenação de Controle de Endemias, Alcinópolis, Mato Grosso do Sul, Brazil [email protected] Keywords: Cave, Fauna, Phlebotomine, vectors. Caves are ecosystems with more constant climatic conditions than those in the external area. Several species of sandflies have their breeding sites in caves, others use them as natural shelters and still others may visit them occasionally. Recently, Alcinópolis municipality in north Mato Grosso do Sul state (MS) has awakened great tourist interest due to the presence of several archaeological sites, including Pitoco cave containing petroglyphs and traces of the ancient inhabitants (of 11,000 years ago) in its proximity. A project is underway to characterize the sandfly fauna of Pitoco cave and its external environment. Monthly captures are being made with CDC light traps installed inside, at the entrance to and in the external environment of the cave in the foothills of the Serra do Bom Sucesso. The data relating to March to May 2014 are here presented and analyzed in the light of other MS cave ecosystems. A total of 1,641 sandflies (49% females and 51% males) of 19 species have been captured, 83.2%, 10.5% and 6.3%, respectively, of them in the interior, at the entrance to and in the external area. Species of four subtribes were found: BRUMPTOMYIINA (4.6%) of 2 species: Brumptomyia avellari and Br. pintoi; SERGENTOMYIINA (14.1%) of 4 species: Deanemyia sp., Micropygomyia acanthopharynx, Mi. quinquefer and Mi. vonatzingeni; LUTZOMYIINA (57.4%) of 6 species: Evandromyia lenti, Ev. saulensis, Lutzomyia cruzi, Lu. longipalpis, Pintomyia christenseni and Sciopemyia sordellii and PSYCHODOPYGINA (23.9%) of 7 species: Martinsmyia oliveirai, Nyssomyia whitmani, Psathyromyia aragaoi, Pa. bigeniculata, Pa. brasiliensis, Pa. campograndensis and Psychodopygus davisi. Lu. dispar, Mt. oliveirai and Mi. quinquefer have been the dominant species (46.4%, 19.7% and 12.7%, respectively). The species of the Longipalpis complex: Lu. cruzi and Lu. longipalpis, visceral leishmaniasis vectors, account for 2.5% of the specimens). Two undescribed females of Deanemyia have been found, this being the first register of this genus in MS. Lu. dispar is a species closely related to Lu. forattinii and Lu. almerioi, both anthropophilic and suspected as vectors of visceral leishmaniasis agent, which are found in other cave ecosystems of MS. So in view of the high density of Lu. dispar in Pitoco cave, its anthropophily, hourly rhythm and potentiality as Leishmania vector call for further investigation in view of the touristic interest of the cave. 132 Sílvia Maria Santos Carvalho, Vanessa Cristina Fitipaldi Veloso Guimarães, Paulo Raimundo Barbosa dos Santos Centro de Pesquisas Aggeu Magalhães, Fiocruz-PE, Brasil [email protected] Keywords: sand flies, Phlebotomine, Lutzomyia longipalpis, Uruçuca Visceral leishmaniasis (VL) is a disease caused by Leishmamia (Leishmania) infantum, which is primarily transmitted by Lutzomyia longipalpis. In Brazil, this species has a wide geographical distribution that coincides with outbreaks of disease, has eclectic and anthropophilic feeding habits and the ability to occupy different ecological niches. Captures were carried out on a farm in Uruçuca municipality, Bahia state, during four consecutive nights of each month using captor of Castro and Center for Disease Control and Prevention light traps from July/2011 to December/2012 distributed in intradomicile, peridomicile (chicken house) and forest remnants from 6:00pm to 6:00am. A total of 151 phlebotomine sand flies belonging to 7 species were captured including 6 specimens of L. longipalpis, one male and four females were found in peridomestic environments and one female in forest remnants. This the first report of L. longipalpis in this region of Bahia state, a traditionally endemic area to American cutaneous leishmaniasis. Although captured sporadically, this finding contributes to knowledge of the geographical distribution of phlebotomine sand flies and highlights the need of monitoring of the area due the risk of emergence of cases VL. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 147-P Ecology & Epidemiology ID 148-P Ecology & Epidemiology THE SAND FLY FAUNA (DIPTERA: PSYCHODIDAE) IN ALTO IVOCHOTE, LA CONVENCIÓN, CUSCO, PERÚ STUDY OF PHLEBOTOMINES IN TWO WILD AREAS OF NORTHEASTERN ARGENTINA AND THE POTENTIAL ASSOCIATION WITH HOWLER MONKEYS (ALOUATTA CARAYA) F Toccas1, D Rado1, Anita Quispe1, C Quispe1, B Galindo1, M Chacon1, MA Quispe-Ricalde2, B Valladares2, R Pacheco1 1 Instituto Universitario de Enfermedades Tropicales y Biomedicina del Cusco – UNSAAC, Perú; 2Instituto Universitario de Enfermedades Tropicales y Salud Publica de Canarias, Universidad de La Laguna, Tenerife, España; 3Instituto de Medicina Tropical Alexander von Humboldt, UPCH, Lima, Perú. [email protected] 133 Mariela F Martínez1,2,4; María S Santini2; María Eugenia Utgés1,2; Martín Kowalewski3,4; Oscar D Salomón1, 4 1 Instituto Nacional de Medicina Tropical, Ministerio de Salud de la Nación, Puerto Iguazú, Misiones, Argentina-REDILA; 2Centro Nacional de Diagnóstico e Investigación en Endemoepidemias, Ministerio de Salud de la Nación, Buenos Aires, Argentina-REDILA; 3Estación Biológica de Corrientes, Museo Argentino de Ciencias Naturales “Bernardino Rivadavia”, Corrientes, Argentina; 4Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina Keywords: Sand Fly Fauna, Alto Ivochote, Cusco [email protected] Alto Ivochote, at 880 m, is an endemic area of leishmaniosis, which cause an important health problem for the people in this province. The objective of this research was to identify and study the sand fly fauna in Alto Ivochote. Sand flies were collected using Shannon traps in extra domestic environments and CDC light traps in intra, peri and extra domestic environments during dry and rainy seasons in 2010 – 2011. Thirty six species of Lutzomyia and Brumptomyia genus were identified. The most frequently species were L. (Nyssomyia) y. yuilli (32.11%), with predominance in extra-domestic environments, and L. (Trichophoromyia) sp. (26.15%) with predominance in intra and peridomestic environments. Activity peaks during 22:00 – 23:00 for L. (N.) y. yuilli and during 04:00 – 05:00 for L. (T.) sp. were observed. There is a statistically significant difference in relation to temperature and presence of L. (T.) sp. (p = 0.021< 0.05) from collections using CDC traps and in relation to humidity and presence of L. (Psychodopygus) h. hirsuta (p = 0.048< 0.05) and L. serrana (p = 0.049< 0.05) from collections using Shannon traps. Keywords: Wild environment; Phlebotomine; Alouatta caraya; vertical strata Support by AECID, Spain. The interaction among the insect vector, the parasite, and the animal reservoir determines the dynamic of infection of vector borne diseases. The animal reservoirs of each Leishmania species vary according to the environment and geographic location, and may include both domestic and wild animal hosts. Sandflies are found in different microenvironments of heterogeneous forest ecosystems. The goal of this study is to describe and compare the Phlebotominae species composition among two forest strata in Alouatta caraya sleeping trees and two wild areas inhabited by this primate. The study sites were Isla Brasilera (IB) Chaco (27°26' S, 59°40' W) and San Cayetano (SC) Corrientes (30°47' S, 55°40' W). The former is a small island in the Paraná River characterized by flooded forest with little to no human contact, and the latter corresponds to the surroundings of a rural locality exhibiting forest patches surrounded by grasslands. In both sites A. caraya (black and gold howler monkey) is the most important arboreal mammal in terms of biomass. Sandflies were captured with light traps hung close to monkey sleeping trees at 1.5 m ('low') and 6-8 m ('canopy') above the ground and set on for two consecutive nights. A total of 964 sandflies were collected in IB (n=7 sleeping trees) belonging to 6 species: Nyssomyia neivai (84.6%), Brumptomyia sp. (12.6%), Migonemyia migonei (2%), Psathyromyia shannoni (0.6%), Ny. whitmani (0.1%), and Evandromyia corteleziisallesi (0.1%). A total of 1125 sandflies were collected in SC (n=9) belonging to 7 species: Ny. neivai (50%), Brumptomyia sp. (25.5%), Mg. migonei (22.8%), Ny. whitmani (0.9%), Ps. shannoni (0.4%), Ev. cortelezii-sallesi (0.3%), and Pintomyia pessoai (0.1%). Ny neivai was the most abundant species in both sites. The comparison of Ny. neivai abundances among sites and heights by nested ANOVA showed significantly higher abundances in the lower stratum (F1,12=6.26; p=0.028), but no significant effect of site factor (F1,12=0.03; p=0.872) or interaction between site and stratum (F1,12=0.31; p=0.588). The presence of potential vectors of Leishmania spp. in the canopy of sleeping trees, though less abundant than in the lower stratum, suggests that A. caraya might serve as wild mammal blood source, and therefore, as an amplifier of the vector population or even a potential reservoir host of leishmanias in these wild areas. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 149-P Ecology & Epidemiology ID 150-P Ecology & Epidemiology PHLEBOTOMINE SANDFLIES (DIPTERA: PSYCHODIDAE) IN A PRESERVED AREA OF THE CAATINGA BIOME IN RIO GRANDE DO NORTE STATE, BRAZIL LUTZOMYIA LONGIPALPIS IN BELLA UNIÓN, DEPARTMENT OF ARTIGAS, URUGUAY. ONE YEAR OF SURVEILLANCE: JULY 2013- JUNE 2014. Maria de Fátima Freire de Melo Ximenes; Renata Antonaci Gama, Tamy Elicia da Silva Lopes, Renato Cesar Melo Freire, Maria de Lima Alves, Marcos Paulo Gomes Pinheiro, José Hilário Tavares da Silva, Marcel Miranda de Medeiros Silva Universidade Federal do Rio Grande do Norte [email protected] Keywords: Phlebotomine sandflies, Caatinga Biome, Lutzomyia longipalpis, preserved forest Phlebotomine sandflies are recognized as Leishmania vectors and differ from other insects of the order Diptera in their morphological and environmental traits. Leishmaniases are classified as neglected diseases by the World Health Organization. In Brazil the number of human cases in their different forms is growing. The Northeast is the primary region of occurrence of visceral leishmaniasis with 47% of the cases. In the state of Rio Grande do Norte cases of Visceral and Cutaneous Leishmaniasis have been reported separately or concomitantly. The visceral form exhibits a higher incidence, reaching a larger number of municipalities. In an attempt at understanding the aspects that contribute to the incidence of disease, in expansion in urban areas of municipalities in the region, a bioecological study of phlebotomine fauna was conducted in the National Forest of Açu (FLONA-Açu), an environment where native Caatinga vegetation is preserved. Captures were made between October 2012 and September 2013 with CDC light traps. A total of 6,577 phlebotomine specimens were captured: Lutzomyia longipalpis (98%), Evandromyia evandroi (1.62%) Evandromyia lenti (0.11%) and Micropygomyia trinidadensis (0.03%). Temperature and relative humidity did not significantly interfere in phlebotomine density in the study period. However, a significant increase in Lutzomyia longipalpis was observed after the rainy season. The high density of L. longipalpis in a preserved environment of the Caatinga demonstrates the importance of constant entomological and epidemiological surveillance by public health sectors, and considering the location of the National Forest in the urban setting of Assú, health education activities are required to help control leishmaniases in dogs and humans and mainly, preserve the natural habitat of phlebotomine species in the Caatinga biome of the National Forest of Açú. 134 1 1 Basmadjián Yester ; Canneva Bruno ; Verger Lorenzo22; 2 3 Vitale Edgardo ; Sequeira Cirino1 ; Lozano Alejandra ; 1 Satragno4 Dinora1, Tort Cecilia ; Viera Ana ; Ríos Cristina ; Lagarmilla Patricia4 and Romero Selva1 1 Departamento de Parasitología y Micología Médica. Instituto de Higiene, Facultad de Medicina, Universidad de la República. Montevideo. Uruguay. Avda. Alfredo Navarro 3051, Montevideo, Uruguay. 2Área de Medicina Preventiva y Epidemiología, Facultad de Veterinaria, Universidad de la República, Montevideo. Uruguay. 3 Liberal Veterinary Professional, Bella Unión, Departamento de Artigas, Uruguay. 4Área de Salud Pública Veterinaria, Facultad de Veterinaria, Universidad de la República. Montevideo. Uruguay. [email protected] Keywords: Lutzomyia longipalpis, Uruguay, Leishmaniasis Visceral Leishmaniasis is a protozoal caused zoonosis that is rapidly expanding in the Southern Cone of the Americas. Even though Uruguay does not integrate the dispersal area of this disease, sporadic cases do occur in neighboring countries (Argentina and Brazil) involving human and canine infections. The city of Bella Unión, with a population of 12,200 habitants (“Instituto Nacional de Estadística” Census 2011) is located in the Artigas Department, on the banks of the Uruguay River, being a mirror town of the argentinian city Monte Caseros (Corrientes province), where since 2009 there is record of dog infected with this disease. In 2010 for the first time in the city of Bella Unión, individuals of Lutzomyia longipalpis, vector of visceral leishmaniasis, were found. No further research was done. From July 2013, a monthly sampling was started in the city, using CDC light traps for two consecutive nights per month. The city was divided into quadrants of 300 meters length, placing the traps in areas that matched the the ""worst case scenario"" definition (abundance of vegetation and presence of dogs among other parameters). Sampling was conducted between July 2013 and June 2014 with a total of 41 Phlebotominae sandflies captured in 6 different quadrants. The species identification was performed by microscopic observation after dissecting and mounting the specimens with Canada balsam all the individuals being identified as Lutzomyia longipalpis. The conducted research highlights the continued presence of the vector in 7 consecutive months (October 2013 to April 2014) with the minimum temperatures between 22°C and 14°C. The persistent finding of L. longipalpis for more than 6 months including lower temperatures than described before by literature must be taken into account. Considering these findings we conclude that the presence of L. longipalpis in the city of Bella Unión has occurred for at least 4 years considering Bella Unión as a vulnerable area. The presence of the vector in much of the city is a matter of concern showing that L. longipalpis has the conditions to live and reproduce. This project was finnanced by the program I+D 2012 of the Comisión Sectorial de Investigación Científica Universidad de la República. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 151-P Ecology & Epidemiology ID 152-P Ecology & Epidemiology PHLEBOTOMINE SAND FLY SURVEY IN CENTRAL AMAZON, AMAZONAS STATE, BRAZIL WITH A NEW REGISTER OF LUTZOMYIA CALIGATA PHLEBOTOMINE SAND FLIES IN A MILITARY TRAINING AREA IN NORTH-EASTERN BRAZIL 1,2 3 2 Oliveira AFJ , Freitas RA , Pessoa, FAC , Aguiar NO 1 Universidade Federal do Amazonas; 2Centro de Pesquisa Leônidas & Maria Deane ILMD – FIOCRUZ AMAZÔNIA, 3Instituto Nacional de Pesquisas da Amazônia 134 155 Abilene Cristina de Arruda Moura, Kamila Gaudêncio da Silva, Débora Elienai de Oliveira Miranda, Miriele Ramalho, Fernando José da Silva, Sinval Pinto Brandão-Filho, Domenico Otranto, Filipe Dantas-Torres 1 [email protected] Departamento de Imunologia, Centro de Pesquisas Aggeu Magalhães, 50670-420 Recife, Pernambuco, Brazil, Dipartimento di Medicina Veterinaria, Università degli Studi di Bari, 70010 Valenzano, Bari, Italy. [email protected] Phlebotomine sandflies are insects of medical importance because some species are vectors of leishmaniasis. There are in Brazil around 400 sand flies species registered and more than 120 of then occur in Amazonas State. The incidence of human cutaneous leishmaniasis cases nearby Manaus and Presidente Figueiredo municipalities are high. Collections of phlebotomine sand flies were done in some forested área of Manaus and Presidente Figueiredo, during different periods of the years of 1998, 2012 and 2013, using CDC light traps and manual collections in tree trunks using a CDC trap as an aspirator. We collected 5.300 individuals, distributed in 12 sub genera, six species groups and 63 species. The species more frequent were L. anduzei, L. davisi, L. flaviscutellata, L. hisurta, L. olmeca nociva, L. squamiventris and L. umbratilis, all of then evolved as a vectors of different species of Leishmania. We collected L. caligata, in Presidente Figueiredo and it is the first occurrence of this species to Amazonas State. Keywords: Sand flies, ecology and Leishmania Sand flies (Phlebotominae) are vectors of several viral, bacterial, and protozoal pathogens, including those of the genus Leishmania, the causative agents of leishmaniasis. The ecology of sand flies is of interest because it may provide background information for a better understanding of the epidemiology of leishmaniasis and to predict the spatial and temporal variations in the risk of leishmaniasis in a given area. The objective of this study was to identify the diversity of species that make up the sand fly fauna in a remnant of Atlantic Forest located in the Campo de Instrução Marechal Newton Cavalcanti (CIMNC), Paudalho municipality, Pernambuco state, north-eastern Brazil. From July 2012 to July 2013, light traps were placed (ca.1.5 m above the ground) in 10 sampling sites (P1-P10) near the burrows of wild animals, trunks and roots of large trees in shady and humid environments. Light traps worked from 18:00 until 6:00 am, for 4 consecutive days each month. The insects captured were initially examined under a stereomicroscope, separated according to sex and kept in 70% ethanol. The specimens were mounted on slides and identified morphologically. A total of 14,403 specimens belonging to 23 species were identified, as follows: Lutzomyia choti(n=10,968), L. longispina(n=1,482), L. sordellii(n=657), L. complexa(n= 650), L. walkeri(n=182),L.amazonensis(n=113),L.wellcomei(n=84),L.evand r o i ( n = 5 4 ) , L . q u i n q u e f e r ( n = 5 2 ) , L . ayrosai(n=48),L.naftalekatzi(n=23),L. barrettoi barrettoi(n=20), L.claustrei(n=20), L. umbratilis(n=16), L.schreiberi(n=11), L.capixaba(n=7), L.brasiliensis(n=4), L.viannamartinsi(n=3), L. whitmani(n=3), L. shannoni(n=3), L. migonei(n=1), L. yuilli pajoti(n=1) and L. oswaldoi(n=1). The number of males (n=7,278) and females (n=7,125) was close to unity. The overall monthly average number of sand flies captured per trap was 110.8, with peaks in March (169.1 sand flies per trap) and April 2013 (332.4 sand flies per trap). This study indicates a high level of sand fly diversity, revealing the occurrence of eight potential vectors of Leishmania spp. in the examined area, including L. complexa, L. amazonensis, L. wellcomei, L. ayrosai, L. umbratilis, L. whitmani, L. shannoni and L. migonei. It also shows a high abundance of sand flies in the study area, with peaks after raining periods. These data will help us to predict the period in which there may be an increased risk of Leishmania transmission in remnants of Atlantic Forest in north-eastern Brazil. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 153-P Ecology & Epidemiology ID 154-P Ecology & Epidemiology R E L AT I N G V E C TO R S P E C I E S P O T E N T I A L DISTRIBUTION AND RICHNESS TO TRANSMISSION FOCI OF CUTANEOUS LEISHMANIASIS IN COLOMBIA RISK OF LEISHMANIOSIS TRANSMISSION IN THE PYRINEAN REGION OF SPAIN ASSESSED THROUGH THE ISOENZYMATIC ANALYSIS OF POTENTIAL SAND FLY VECTORS (PHLEBOTOMUS ARIASI AND P. PERNICIOSUS) Cristina Ferro, Marla Lopez, Patricia Fuya, Ligia Lugo, Juan Manuel Cordovez, Camila Gonzalez Universidad de los Andes, Instituto Nacional de Salud 135 Ballart C1,2; Pesson B3; Martín-Sánchez J4; Alves-Pires C5; Morillas F4; Afonso MO5; Portús M1; Gállego M1,2 1 [email protected] Keywords: Phlebotominae, distribution, eco-epidemiology, Leishmaniasis Laboratori de Parasitologia, Facultat de Farmàcia, Universitat de Barcelona (UB), Spain; 2Centre de Recerca en Salud Internacional de Barcelona (CRESIB), Hospital Cínic-UB, Spain; 3Laboratoire de Parasitologie, Faculté de Pharmacie, Université de Strasbourg and , France; 4Departamento de Parasitología, Facultad de Farmacia, Universidad de Granada, Spain; 5Unidade de Parasitologia e Microbiologia Médicas (UPMM), Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Lisboa, Portugal [email protected] Leishmaniasis is a complex parasitic disease with worldwide distribution, caused by various species of the genus Leishmania related to different clinical manifestations. In America, Leishmania is transmitted by vector insects of the genus Lutzomyia, who maintain the enzootic cycle in distinct regions by circulating between sylvatic and domestic mammals, as well as humans who enter the cycles as accidental reservoirs. In Colombia, Leishmaniasis is an endemic disease with 95% of all cases being cutaneous. Cases are reported in several regions of the country with diverse ecological characteristics, nevertheless, factors like displacement of human populations from endemic areas, and the intervention of sylvatic areas by the introduction of agriculture and other socioeconomic and environmental factors seem to have an impact on the rearrangement of new transmission cycles. Our study aimed to analyze the relationship between vectors' distribution and the number of cases reported of Cutaneous Leishmaniasis (CL) in Colombia. Nineteen sand fly species reported as of medical importance for the transmission of CL in Colombia were included and ecological niche modeling was performed to explore the potential distribution of vector species in Colombia and their relation to transmission areas and ecosystems. In total, 560 single records were obtained with L. gomezi as the species with the highest number of records and N. ylephiletor y P. carrerai thula with the lowest. Lutzomyia gomezi and P. panamensis had the widest distribution in contrast with P. spinicrassa, P. youngi, and P. nuneztovari which was more restricted to a specific region. Potential species' richness was calculated and overlapped to the average reported cases from 2005-2013 in Colombia. The departments with the highest number of cases reported corresponded to Antioquia, Nariño, Tolima, Meta, Santander and Bolivar. The results showed that areas with highest prevalence have 2-4 different vector species predicted as present. We were able to conclude that distribution of vectors with medical importance in Colombia correspond to disturbed areas augmenting the domestication potential; vector species with similar distribution are affected by the same climatic variables, and finally, we highlight the importance of the use of distribution maps as a tool for the development of strategies for prevention and control of diseases. Keywords: Phlebotomus ariasi, P. perniciosus, Isoelectrofocusing, leishmaniosis risk Leishmaniosis, human and canine, does not have a uniform distribution in Spain, and some northern regions are not considered endemic. Nevertheless, in the last years, cases of canine leishmaniosis have been found in some of these northern areas, for example, in the Pyrenean region. Leishmaniosis transmission is highly specialized, with sand flies acting as vectors. In Spain, only the females of two species, Phlebotomus perniciosus and P. ariasi, are confirmed vectors of Leishmania infantum.. Besides morphological identification, several new techniques are being applied in the study of sand flies. These techniques distinguish between cryptic species and populations with different vectorial capacity, and are therefore useful in entomological and epidemiological research on leishmaniosis. The application of the new technology assists in the risk of leishmaniosis transmission in new areas. In this work, we applied isoelectric focusing (IEF) to study sandflies captured in northern Spain. The sand flies (522 P. ariasi and 37 P. perniciosus) come from two Pyrenean counties of Lleida province (Catalonia): Pallars Jussà (170 specimens) and Pallars Sobirà (389). An IEF analysis using eight enzyme loci was carried out (PGI, PGM, HK, FUM, ACO, MDH1, MDH2 and 6PGD). The results were compared with those of other sand fly populations of leishmaniosis-endemic areas in Spain, France and Portugal. In the case of P. ariasi, the specimens were grouped with populations from southern France (Aston and Vira), while two other distinct groups were found, one consisting of populations from Cheires (Alto Douro, Portugal) and Rio Tinto (Huelva, Spain) and the other group from the French towns of Les Tourrettes (La Drôme Departement) and Roquedur (Gard Departement). Similar results were found in the case of P. perniciosus, the Pyrenean populations of Lleida being grouped with others from a highly leishmaniosisendemic area of southern Catalonia (Priorat, Tarragona) and the east of France (Drôme), while the population of southwestern Spain (Huelva) has a greater similarity to the one of Portugal. The results show that sand fly populations of Lleida province in northern Spain, where leishmaniosis is poorly studied and was considered non endemic, resemble those of other areas of Spain and France where the disease is endemic. Autochthonous cases of canine leishmaniosis were recently diagnosed in this area. Financial support: Spanish Ministerio de Ciencia e Innovación (CG12010-22368-C02-01). N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 155-P Ecology & Epidemiology ID 156-P Ecology & Epidemiology IRRIGATION IN THE ARID REGIONS OF TUNISIA IMPACTS THE ABUNDANCE AND APPARENT DENSITY OF SAND FLY VECTORS OF LEISHMANIA INFANTUM SANDFLIES (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) FROM THE NISIA FLORESTA NATIONAL FOREST, METROPOLITAN REGION NATAL, RIO GRANDE DO NORTE STATE, BRAZIL Walid Barhoumi1, Whitney A. Qualls2, Reginald Archer3, Douglas O Fuller3, Ifhem Chelbi1, Saifedine Cherni1, Mohamed Derbali1, Kristopher L Arheart2, Elyes Zhioua1 and John C Beier2 Marcos Paulo Gomes Pinheiro, Maria de Fátima Freire de Melo Ximenes, Maria de Lima Alves, João Batista da Silva Júnior, José Hilário Tavares da Silva, Marcel Miranda de Medeiros Silva, Tamy Elicia da Silva Lopes 1 Laboratory of Vector Ecology, Pasteur Institute of Tunis, 13 Place Pasteur BP 74, 1002 Tunis, Tunisia; 2Department of Public Health Sciences, University of Miami Miller School of Medicine, Miami, Florida, United States of America; 3Department of Geography and Regional Studies, University of Miami, Miami, Florida, United States of America [email protected] Keywords: Human visceral leishmaniasis, Phlebotomus perfiliewi, integrated vector management, remote sensing approaches The distribution expansion of important human visceral leishmaniasis (HVL) and sporadic cutaneous leishmaniasis (SCL) vector species, Phlebotomus perfiliewi and P. perniciosus, throughout central Tunisia is a major public health concern. This study was designed to investigate if the expansion of irrigation influences the abundance of sand fly species potentially involved in the transmission of HVL and SCL located in arid bioclimatic regions. Geographic and remote sensing approaches were used to predict the density of visceral leishmaniasis vectors in Tunisia. Entomological investigations were performed in the governorate of Sidi Bouzid, located in the arid bioclimatic region of Tunisia. In 2012, sand flies were collected by CDC light traps located at 9 irrigated and 9 non-irrigated sites to determine species abundance. Eight species in two genera were collected. Among sand flies of the subgenus Larroussius, P. perfiliewi was the only species collected significantly more in irrigated areas. Trap data were then used to develop Poisson regression models to map the apparent density of important sand fly species as a function of different environmental covariates including climate and vegetation density. The density of P. perfiliewi is predicted to be moderately high in the arid regions. These results highlight that the abundance of P. perfiliewi is associated with the development of irrigated areas and suggests that the expansion of this species will continue to more arid areas of the country as irrigation sites continue to be developed in the region. The continued increase in irrigated areas in the Middle East and North Africa region is of high importance as it is associated with the spread of L. infantum vector P. perfiliewi. Integrated vector management strategies targeting irrigation structures to reduce sand fly vector populations should be evaluated in light of these findings. 136 Universidade Federal do Rio Grande do Norte [email protected] Keywords: Phlebotomine sandflies, Leishmaniasis, Atlantic Forest. Phlebotomine sandflies, important insects in the medical-veterinary field, are vectors of Leishmania protozoa, which cause visceral and cutaneous leishmaniasis, serious diseases if left untreated, and widely distributed in Brazil. The present study aimed at assessing the composition, abundance and diversity, as well as the relationship with climatic factors of leishmaniasis-transmitting phlebotomine sandflies, in the Nisia Floresta National Forest (FLONA). This FLONA, a conservation and research unit covered by different plants exotic and native to the Atlantic Forest, is situated in the metropolitan region of Natal, Rio Grande do Norte state, Northeastern Brazil. Two monthly captures were made over a one-year period, from May 2012 to April 2013, in CDC light traps placed at four points, two more anthropized, near the occupied area of FLONA, one in an Atlantic Forest area, and the other in the Pinus forest located within this area. A total of 2,999 specimens of ten phlebotomine sandfly species were captured as follows: Evandromyia walkeri, Evandromyia evandroi, Psychodopygus wellcomei, Sciopemyia sordellii, Psathyromyia brasiliensis, Lutzomyia longipalpis, Evandromyia lenti, Psathyromyia shannoni, Nyssomyia whitmani and Nyssomyia intermedia. The area with greatest sandfly abundance was the Atlantic Forest (32.4%), followed by the bamboo plantation (26.3%), and with greatest diversity in the Pinus forest (H'=1.097), followed by the Atlantic Forest (H'=1.008). The most abundant species was E. walkeri, with 76% (SISA=1.0), followed by E. evandroi with 16.5% (SISA=0.87). It is important to underscore the presence of L. longipalpis, vector species of L. infantum, with 0.64% (SISA=0.80), as well as P. wellcomei with 1.8% (SISA=0.82), N. intermedia with 0.03% (SISA=0.04) and N. whitmani with 0.10% (SISA=0.04), species present in cycles of cutaneous leishmaniasis in different regions of the country. In general phlebotomine sandflies were more abundant in the rainy season. L. longipalpis was more adapted to environments with greater anthropic intervention, since it represented 79% of the sandflies captured, while P. wellcomei was associated to more preserved areas, with 80% in this type of environment and the rainy season. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 157-P Ecology & Epidemiology ID 158-P Ecology & Epidemiology ENTOMOLOGICAL SURVEY OF PHLEBOTOMINE SAND FLIES (DIPTERA: PSYCHODIDAE) IN TURKEY DISTRIBUTION OF SANDFLIES SPECIES IN A LOCALITY ENDEMIC FOR AMERICAN TEGUMENTARY LEISHMANIASIS FROM THE NORTH OF ARGENTINA, BORDERING WITH BOLIVIA Ozge Erisoz Kasap, Bulent Alten 137 Department of Biology, Hacettepe University, Ankara, Turkey [email protected] Keywords: distribution, sand flies, Turkey In Turkey, visceral and canine leishmaniasis caused by Leishmania infantum are endemic along the Aegean and Mediterranean costs while cutaneous leishmaniasis caused by Leishmania tropica and Leishmania infantum is endemic in the south and southeast of Anatolia. Hence, most of the sand fly sampling efforts has been focused on these regions to determine the possible vector species so far. However, there is no detailed data of the sand fly fauna of the regions where no leishmaniasis or other sand fly borne diseases cases have never been reported. In this work we present the results of an intensive entomological survey carried out in Turkey. Sand fly samplings were conducted in 19 provinces covering a wide range of area including the Black Sea, Thrace, Aegean, Southern, Eastern and Central regions of Turkey between 2005 and 2013. Sand flies were found to have a wide geographical and altitudinal distribution ranging from 36 and 1410 meters above sea level. A total of 3846 specimens belonging to 12 species of Phlebotomus and two of Sergentomyia were collected. P. perfiliewi s.l. (23.17 %), P. major s.l (23.01 %), and P. tobbi (16.23 %) composed the most the sand flies collected. P. papatasi, P. alexandri, P. sergenti, P. balcanicus, P. halepensis, P. simici, P. burneyi, P. kandelakii, Transphlebotomus sp., S. dentata and S. minuta were recorded as the other species found in the sampling locations. Larroussius species which are the proven or probable vectors of L. infantum in the Mediterranean region were found to have the widest distribution. Finding these species in almost all the sampling locations should be considered as a risk factor for the spreading of the leishmaniasis in Turkey. Copa GN 1,3, Marco JD 3,5, Nasser JR 1,2 Gil JF 1,2,3,4 1 Instituto de Investigaciones en Enfermedades Tropicales (IIET), Sede Regional Orán, Facultad de Ciencias de la Salud, Universidad Nacional de Salta (UNSa). 2Cátedra de Química Biológica y Biología Molecular, Facultad de Ciencias Naturales, (UNSa). 3 Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET). 4 Instituto de Investigación en Energías No Convencionales, Departamento de Física, (UNSa). 5Instituto de Patología Experimental, Facultad de Ciencias de la Salud, (UNSa). [email protected] Keywords: Sandflies, American tegumentary leishmaniasis, Aguas Blancas, distribution Some sandflies species can vectorize Leishmania protozoan parasites which cause America tegumentary leishmaniasis (AtL). This disease is endemic in Argentina, with high incidence in department of Oran, Salta Province. Previous studies have reported the presence of sand flies in environments that include wilderness areas, secondary vegetation, rural, peri-urban and urban areas. The Aguas Blancas, city located in Oran, is scenary of a fluid transit of people between Argentina and Bolivia. The aim of this study was to analyze and describe the abundance and geographic sandflies distribution in Aguas Blancas, border locality of the north of Argentina. The study was conducted in October 2013 in nine sampling sites, which corresponded to residential courtyards widely distributed in the town. The capture of sandflies was performed by CDC traps placed three consecutive nights from 19:00 pm to 7:00 pm. The distribution maps were generated using ArcGIS 9.2 software and statistical analysis was performed using the chi-square test. A total of 230 specimens were captured: cortelezzii complex (30%), Evandromyia sallesi (28.26%), Nyssomyia neivai (24.35%), Mygonemyia migonei (16.52%). The 12.87% of the females were in a gravid state. The sites with higher sandflies abundance were located at the edges of the city, decreasing to zero in the center (p<0.0001). The cortelezzii complex was the most abundant and showed a wider geographical distribution that Ny. neivai (p<0.001). Thus, this is the first report for rain forest region, in which Ny. neivai there is not found as the dominant specie. Sandflies species found during this study are suspected of transmitting AtL. Since Aguas Blancas is an area with constant flow of persons for a migration process through the customs office located in the city. Thus, it is important to consider the risk of transmission of this disease specially when considering the potential exposure of nonimmune population from different places of Argentina and Bolivia. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 159-P Ecology & Epidemiology ID 160-P Ecology & Epidemiology EFFECTIVENESS OF A RAPID MULTIDISCIPLINARY P R O TO C O L F O R T H E E P I D E M I O L O G I C A L E VA L U AT I O N O F A N A C T I V E F O C U S O F C U TA N E OU S L E I S HMA N I A S I S : R E ME D I OS , ANTIOQUIA, COLOMBIA ABUNDANCE ASSOCIATION OF NYSSOMYIA WHITMANI WITH THE NUMBER OF FARM ANIMALS AND ITS DWELLING DISTANCE TO THE HOUSE, IN AREAS WITH TEGUMENTARY LEISHMANIASIS TRANSMISSION IN PUERTO IGUAZÚ, MISIONES, ARGENTINA Freddy Ruiz-Lopez1, Horacio Cadena1, Andrés Vélez Mira1, Laura Posada-López 1,2, Angelica Contreras1,2,;Didier Tirado Duarte1, Iván Darío Vélez1 1 PECET Programa de Estudio y Control de Enfermedades Tropicales, Universidad de Antioquia, Medellín, Colombia. 2GSM Grupo de Investigación en Sistemática Molecular, Universidad Nacional sede Medellín, Colombia. 138 Villarquide ML1,2; Fernández MS3,4; Santini MS2; Salomón OD3,4 1 Fundación H.A. Barceló;2 Centro Nacional de Diagnóstico e Investigación de Endemoepidemias-REDILA;3Consejo Nacional de Investigaciones Científicas y Técnicas;4Instituto Nacional de Medicina Tropical- REDILA [email protected] [email protected] Keywords: cutaneous leishmaniasis, epidemiology, Lutzomyia. Keywords: Tegumentary Leishmaniasis, Leishmania braziliensis, Nyssomyia whitmani. Leishmaniasis (L) is a group of infectious diseases transmitted by the bite of female insects belonging to the genus Phlebotomus and Lutzomyia. In humans leishmaniasis can affect the skin (cutaneous L), mucous membranes (mucosal L) and internal organs (visceral L). In recent decades human migration, unplanned urbanization, mining and climate change have facilitated the adaptation of insect vectors and wild mammals to peridomestic habitats causing human epidemic outbreaks in urban and suburban areas. The aim of this research was to evaluate the epidemiology of an active focus of cutaneous L in Colombia by optimizing and applying the protocol described by the National Institute of Health of Colombia. A rapid protocol was developed by this study for implementation over a period not exceeding five days. The methods included: 1 Ecoepidemiological surveys; 2 Blood sampling of patient; 3 Capture and sampling in mammals; 4 Application of the Montenegro skin test; 5 Sand fly collection; 6 Laboratory tests; 7 Identification of Lutzomyia species. Thirty five homes were visited and blood samples were taken from eight people with suspicious skin lesions. Of these, seven were positive by direct smear and the parasite was isolated in six of these samples. Restriction Fragment Length Polymorphism (RFLP) identified Leishmania panamensis as the circulating strain in this region. A total of 75 sand flies were collected that grouped into nine species including Lu. gomezi, Lu. panamensis, Lu. trapidoi and Lu. yuilli yuilli, all of which are recognized Leishmania vectors in Colombia. Five mammals were captured, but none were found to be infected. The area studied is characterized by unplanned human settlements located on village outskirts and is surrounded by clusters of forest, where the main source of work is the extraction of gold in a mining shaft. People are infected due to the close proximity of their homes to wooded areas that promote the natural transmission cycle of leishmaniasis. The presence of Leishmania vectors, wild mammal intermediate hosts and human settlements is indicative of high risk environments for cutaneous L transmission. These results show that short studies of endemic foci, addressing all important components in the transmission cycle of the disease, might be sufficient for the epidemiological evaluation of regions with active cases of cutaneous L. For this a multidisciplinary team is required to address all the components of the transmission cycle. This study was performed in Puerto Iguazú, Misiones, Argentina, in an area of recent deforestation-human settlement called '2000 hectáreas'. Tegumentary leishmaniasis (TL) is recorded there since the 2004-2005 outbreak. Leishmania braziliensis was the circulating parasite and Nyssomyia whitmani the incriminated vector. The aim of this work was to study the association between the abundance of Ny. whitmani and the amount of breed animals (chickens and pigs), and the vector abundance related to the distance between the house and the animal dwellings or the forest edge. To develop this study 17 farms were selected. In each farm, Phlebotominae abundance was measured in the house, henhouse, pigsty and forest edge (sites). Captures were done at each site using mini CDC traps on one night during February 2010 (5 pm to 8 am). Number of chickens and pigs, and distance from sites to the house were recorded for each farm. Similarity between house, henhouse and forest edge was calculated with the Sörensen index. Linear regression models for Ny. whitmani´s abundance were constructed with environmental characteristics as explanatory variables. Abundance at all the same sites added of Ny. whitmani per site was: house, 263; henhouse, 3675; pigsty, 1392; and forest edge, 1546. The highest similarity between sites was seen between henhouse and forest edge (Is=0.39). The abundance of Ny. whitmani was found to be positively related with number of chickens and pigs (R2=0.25; SE=0.03; CI95%= -0.01, 0.13; R2=0.47; SE=0.22; CI95%= -0.03, 1.02, respectively). Forest edge was positively associated to number of chickens and negatively with distance to the house (R2=0.83; SE=0.01; CI95%= -0.003, 0.045; SE=0.006; CI95%= -0.047, -0.019, respectively). Therefore, in scenarios of recent deforestation/human settlement in Northeastern Argentina, the likelihood of human-Ny whitmani contact increases with the breeding of pigs or chickens, that attracts vectors from sand fly source populations located in the forest or the domestic area. Further, the risk of transmission is related to the abundance of animals, and the spatial distribution of the animal dwellings, human sleeping areas and the edge of the forest. These results suggest that the strategies to prevent TL in the study area, in human settlements close to the edge of the forest, should take into account the practices associated with animal management, and the habits related to the use of the domestic space (peridomestic and domestic). N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 161-P Ecology & Epidemiology ID 162-P Ecology & Epidemiology BIODIVERSITY OF PHLEBOTOMINAE SUBFAMILY (DIPTERA: PSYCHODIDAE) IN THE COLOMBIAN AMAZON AND ORINOCO S A N D F LY V E C T O R S O F C U TA N E O U S LEISHMANIASIS ON THE WESTERN SLOPES OF THE ECUADORIAN ANDES Cristina Ferro, Patricia Fuya, Ligia Lugo, Camila González Sandra Enriquez1, Jazzmin Arrivillaga1,5, Franklin Vaca1, Washington Benitez Ortiz1, Oscar Kirstein2, Ibrahim Abassi2, Manuel Calvopiña3, Katherine Pozo1, Vanessa Romero1, Orlando Chiluisa1, Ernesto Villacrés1 and Alon Warburg 1,2,4 Instituto Nacional de Salud (Laboratorio de Entomología-LNR), Universidad de Los Andes (CIMPAT) [email protected] Keywords: Phlebotominae, Biodiversity, Amazonia and Orinoquia species The Phlebotominae subfamily, with more than 800 species, known for its biodiversity and the role played by some of its members as disease vectors, is widely distributed in our planet. In the New World Phlebotominae specimens from about 512 species have been recorded from Canada to Argentina, being the Neotropical countries the ones with the highest number of species. Among these Colombia with 163 species, is second after Brazil, with 242. In order to update the information about Phlebotominae distribution in the Colombian Orinoquian and Amazon region, records of species by locality were obtained from collections made since the early sixties, and classified according to Galati, 2003. Data from103 locations were collected with an overall record of 92 species, 57 of them with exclusive presence on the Amazon and/or Orinoquian region. The number of species per genus was Bichromomyia (3), Brumptomyia (2), Evandromyia (8), Lutzomyia (8), Micropygomyia (8), Migonemyia (1), Nyssomyia (5), Pintomyia (6), Pressatia (4), Psathyromyia (15), Psychodopygus (15), Sciopemyia (4), Trichophoromyia (9), Trichophygomyia (5), Viannamyia (2). The species with the widest distribution was N. antunesi. Among the species with epidemiological importance in leishmaniasis we have N. umbratilis, N. yuilli, N. antunesi, B. flaviscutellata, B. olmeca bicolor, B. reducta, P. davisi and P. hirsutus; the first three with high abundance, anthropophilic behavior and presence within the households. In conclusion, Phlebotominae biodiversity in the Orinoquian and Amazon regions of Colombia is high. It adds 57 species to the total list of species recorded in Colombia and shares 102 taxa with records from other regions in the country, some of them located in the genus Nyssomyia, Bichromomyia Psychodopygus with vectorial background, mainly in cutaneous leishmaniasis where this clinical form represents 23% of cases in the country. 139 1 Centro Internacional de Zoonosis (CIZ) Universidad Central del Ecuador, Quito, Ecuador, 2The Kuvin Center for the Study of Infectious and Tropical Diseases, Department of Microbiology and Molecular Genetics, The Institute of Medical Research Israel-Canada, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem, Israel. 3Unit of Molecular Parasitology &Tropical Medicine, Centro de Biomedicina, School of Medicine, Universidad Central del Ecuador, Quito-Ecuador. 4 Prometeo, Secretaría Nacional de Educacion Superior, Ciencia, Tecnologia e Innovacion (SENESCYT), Ecuador. 5Laboratorio Genética de Poblaciones, Sección Invertebrados, Dept. de Estudios Ambientales, Univ. Simón Bolívar, Venezuela; [email protected] Keywords: Cutaneous Leishmaniasis, Lutzomyia trapidoi, Leishmania naiffi In Ecuador cutaneous leishmaniasis (CL) is endemic and common in most regions of the country including the western slopes of the Andes, Andean valleys and the Amazonian lowlands. Several Leishmania spp. cause CL in Ecuador including L.(Viannia) guyanensis, L. (V.) panamensis, L. (V.) brazilensis, L. mexicana and L. major-like parasites. We are conducting a project designed to study and characterize the epidemiology of CL in several communities in the district of Pichincha on the Western slopes of the Ecuadorian Andes, where increasing numbers of CL cases are being reported. Although the main aim of the project is to optimize diagnosis and treatment of human CL cases by local clinics, we are also performing entomological studies with the aim of devising disease prevention strategies through sand fly control and curtailment of human-sand fly contact. Here we report preliminary findings on sand fly species altitudinal distribution, Leishmania infections and blood source identification of sand flies in and near rural communities. The most common sand fly was Lutzomyia trapidoi which was found in all altitudes (200-1700 m above sea level) and habitats. The blood hosts of 16 females were successfully identified using cytochrome b PCR followed by DNA sequencing: 13 human, 2 opossum (probably Didelphis marsupialis), 1 Porcupine (Coendou quichua). In parallel, Leishmania infections were detected using ITS1 PCR followed by DNA sequencing. Leishmania DNA was positively identified in 9 females (6 Lu. trapidoi, 2 Lu. panamensis, 1 Lu. hartmanni) out of 77 examined which is a very high infection rate. All DNA sequences were of Leishmania (Viannia) spp. including L.(V.) braziliensis, L.(V.) panamensis, L.(V.) guyanesis and L.(V.) naiffi. This is the first record of L.(V.) naiffi from the western part of Ecuador. Studies are ongoing and more data will accrue in the near future. Acknowledgements: This work was funded by Fundação de Assistência Médica Internacional (AMI), Portugal and Prometeo Project of the Secretaria Nacional de Educacion Superior, Ciencia,Tecnologia e Innovacion (SENESCYT), Ecuador. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 163-P Ecology & Epidemiology ID 164-P Ecology & Epidemiology ABUNDANCE OF PHLEBOTOMINAE IN ENDEMIC AREA OF CUTANEOUS LEISHMANIASIS, TUCUMÁN, ARGENTINA PHLEBOTOMINE SANDFLIES (DIPTERA: PSYCHODIDAE) OF MOROCCO: RESULTS OF E N TO M O L O G I C A L S U RV E Y S A L O N G T H E O U A R Z A Z AT- M ' H A M I D , Z A G O R A CHEFCHAOUEN-NADOR AND ZAGORA-TATAMARRAKESH ROADS Direni-Mancini JM, Fuenzalida AD, Saracho-Bottero MN, Lizarralde de Grosso MS, Salomón OD, Quintana MG Instituto Superior de Entomología – UNT, REDILA [email protected] Keywords: Migonemyia migonei, Nyssomyia neivai, Evandromyia cortelezzii, meteorological variables 140 Ouanaimi Fouad1; Boussaa Samia1,2; Kahime Kholoud1; Echchakery Mohamed1; Boumezzough Ali1 Equipe d'Ecologie Animale-Environnement, Faculté des Sciences Semlalia, Université Cadi Ayyad, Marrakech, Maroc; 2ISPITS-Institut Supérieur des Professions Infirmières et des Techniques de Santé, Ministère de Santé, Marrakech, Morocco [email protected] In Argentina, 30 species of sandflies were recorded, which belong to 13 genera. Cutaneous leishmaniasis (CL) cases have increased gradually in the province of Tucumán as well as in the remaining endemic region. The last two outbreaks took place in 2003 and 2004. Nyssomyia neivai is the main species incriminated in the transmission of Leishmania braziliensis, parasite responsible of CL in the northwest region of Argentina. The aims of this research were to compare the abundance of Phlebotominae species in different sites and seasons in addition to determine the meteorological variables that have influenced in the abundance of the species in the south of the province (27º35´05.89´´S - 65º37´11.70´´W).The study area belongs to Yungas rainforest region, which has been currently modified for different culture types. Five sample sites were selected based on entomological and epidemiological antecedents. The sandflies were captured with CDC light traps monthly for two consecutive days from July 2008 to July 2009. The followings meteorological variables were registered on the day of capture: Temperature (TºC), Relative humidity (Rh%), Precipitation (Pp), Velocity of wind (Vw) and Maximum velocity of wind (MVw). A total of 6793 individuals were captured belonging to the species Migonemyia migonei (50.07%), Ny. neivai (49.20%) and Evandromyia cortelezzii (0.74%). The abundances of the three species were significantly different. Three out five sites were the most abundant. Summer was the most abundant season for the three species, followed by fall, spring and winter. The main variables which affected the abundance of Mg. migonei were the TºC and MVw (R2= 0.87); for Ny. neivai were the Rh% and MVw (R2= 0.75) and for Ev. cortelezzii was the TºC (R2= 0.45). Mg. migonei was more abundante in summer and spring (highest seasons of activity for sandflies) whereas Ny. neivai, primary vector, was more abundant in fall which is the highest transmission season. The three most abundant sites are located at the outbreak area of 2003, where CL cases were concentrated. The information provided in this study contributed to the knowledge of the distribution of abundance in time and space of CL vectors in northwestern Argentina. Keywords: Sand flies, transects, entomological surveys, Morocco Phlebotomine sand flies (Diptera: Psychodidae) are the vectors for leishmaniasis as well as for arboviruses and bartonellosis. Leishmaniasis are endemic in Morocco with two described forms (visceral and cutaneous). Knowledge of vectors populations' distribution is important in predicting the spatial variations in the risk of disease. Previous studies in Morocco showed that the distribution of sand flies was due, in great part, to the bioclimate. Current findings in Morocco showed that altitude (through the gradient on temperature, pressure and precipitation) and aspect (through climate and vegetation) have an influence upon the spatial distribution and density of the sand fly fauna. In the present work, we discuss the possible effect of many ecological factors on the diversity and distribution of sand flies in southeastern Morocco along three transects (Ouarzazat-Mhamid; Zagora-Chefchaouen-Nador and Zagora-Tata-Marrakesh). We give a particular attention to ZCL foci (Ouarzazat, Zagora, Tata and Errachidia) with the aim to update their entomological data. Among the 2056 sandflies collected, in June 2010, on the Ouarzazat-M'hamid road, Phlebotomus longicuspis was the most common species (25.5%), followed by P. papatasi (23.8%) Sergentomyia fallax (12.6%), P. sergenti (16.6%), S. minuta (11%), P. bergeroti (5.4%), P. dreyfussi (1.8%), P. christophersi (1.7%) and P. ariasi, P. perniciosus, P. alexandri, P. chabaudi and P. chadlii with less than 1% each. On the Zagora-Tata-Marrakesh road, P. papatasi was the most prevalent species (24.2%) of the 2425sandflies that were collected in June 2011, followed by P. longicuspis (17.3%), S. minuta (16.4%), P. sergenti (11.9%), S. fallax (5.2%), P. dreyfussi (3.1%), P. christophersi (2.8%), P. bergeroti (2.6%) and P. alexandri (1.2%). We collected as well P. africana, P. chabaudi, P. ariasi and P. chadlii with less than 1% each. In June 2012, 2828 sandflies were collected on the Zagora-Chefchaouen-Nador road. P. papatasi was the most prevalent species (33.5%), followed by P. sergenti (25.2%), P. longicuspis (15.1%), P. perniciosus (6%), S. fallax (5.7%), S. minuta (4.7%), P. alexandri (2.5%), P. chadlii (1.7%), P. bergeroti (1.5%) and P. dreyfussi (1.5%). S. Africana, P. christophersi, P. chabaudi and P. ariasi with less than 1% each. The distribution of potential vectors along the three transects, according to altitude and bioclimate, was explored. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 141 ID 165-P Ecology & Epidemiology ID 166-P Ecology & Epidemiology S PAT I A L R E L AT I O N S H I P B E T W E E N ENVIRONMENTAL FACTORS AND SAND flY (DIPTERA: PSYCHODIDAE) DISTRIBUTION ASSOCIATED WITH LEISHMANIASIS RISK IN CENTRE & SOUTHERN MOROCCO IDENTIFICATION AND CHARACTERIZATION OF LARVAL LODGING OF PHLEBOTOMINE SAND FLIES (DIPTERA : PSYCHODIDAE) IN MARRAKECH, MOROCCO Boussaa Samia1, 2; Ouanaimi Fouad1; Boumezzough Ali1 Kahime K, Boussaa S, Ouanaimi F & Boumezzough A Faculté des Sciences Semlalia, Université Cadi Ayyad, Marrakech, Morocco 1 Equipe d'Ecologie Animale-Environnement, Faculté des Sciences Semlalia, Université Cadi Ayyad, Marrakech, Maroc; 2ISPITS-Institut Supérieur des Professions Infirmières et des Techniques de Santé, Ministère de Santé, Marrakech, Morocco [email protected] [email protected] Keywords: SIG, Leishmaniasis, sand fly, risk, Morocco Leishmaniasis, a highly neglected disease, currently presents a significant health problem throughout Africa. In Moroccan context, both cutaneous (CL) and visceral leishmaniasis (VL) have been reported. CL is caused by three clinically important Leishmania species (L. major, L. tropica, and L. infantum), a flagellate protozoa of the family Trypanosomitidae, while, VL is caused by L. infantum. Recently, the Mucocutaneous form was also reported but remains rare. Leishmania infection is transmitted to human host as a result of a bite by an infected female sand fly (Diptera: Psychodidae, Phlebotominae) of the genus Phlebotomus. Over the past decade, the epidemiological situation of CL has significantly increased with its geographic expansion to previously free areas and the emergence of overlapping foci of CL and VL in several provinces of Morocco. In order to determine Leishmaniasis risk membership in central- south Morocco, Geographic Information System (GIS) was used to investigate the current spatial distributions of sand fly-borne pathogen risk. The study was conducted in 69 sites where altitude varies from 14 to 2123 m. These sites belong to two administrative areas, the region of Marrakech-Tensift- Al Haouz and the region of Souss-Massa-Draa. In both regions, climate and vegetation are typically Mediterranean, with hot, dry summers and maximum rainfall recorded in autumn and spring. By sticky traps, we collected a total of 15 313 specimens, belonging to 15 species of which10 species of the genus Phlebotomus (57,38%). Proven and suspected Leishmania vectors were well presented, in our study area, such Phlebotomus (Larroussius) perniciosus Newstead (14,35%), Phlebotomus (Phlebotomus) papatasi Scopoli (14,06%), Phlebotomus (Paraphlebotomus) sergenti Parrot (12,85%), Phlebotomus (L.) longicuspis Nitzulescu (10,74%) and Phlebotomus (L.) ariasi Tonnoir (2,68%). Sand fly repartition linking to contextual environmental factors including geology, soil, vegetation and climate were discussed for each species. Keywords: sand flies, un-rotated genitalia, larval lodging, Marrakech Phlebotomine sand flies are the only vectors known of leishmaniasis. These diseases are a serious increasing public health problem in Morocco. In the aim to identify and characterize sand fly breeding habitats in Marrakech city, our entomological investigations were carried out during one year study in two sites (Akioud and Semlalia). Using sticky paper, specimens were collected continuously between October 2002 and September 2003 in Akioud, and from July 2002 to July 2003 in Semlalia. Newly eclosed male sand flies were detected based on their un-rotated genitalia. A total of 3571 specimens was trapped during this entomological survey. Phlebotomus papatasi was the most abundant species (53,2%), followed by Sergentomyia minuta (20,1%), S. fallax (11,5%), P. sergenti (10,7%) and P. longicuspis (4,5%). The sex ratio was in favour of males for all species with the exception of P. sergenti (61.4% of females) in Akioud and P. longicuspis (50% of females) in Semlalia. In Marrakech, breeding habitats were identified for three, of five collected species, as well as their emergence phases. Our results show that Akioud can be breeding habitats for two Sergentomyia species (S. minuta and S. fallax) while, Semlalia was identified as breeding habitats for P. sergenti.Soil sample was collected from both sites where newly eclosed male sand flies were detected. These samples were used to determine organic matter values, humidity and soil texture of each larval lodging. Characterization of these soil samples shows some ecological requirements of immature stages development for these sand fly species. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 167-P Ecology & Epidemiology ID 168-P Ecology & Epidemiology LEISHMANIASIS IN MOROCCO: STILL A TOPICAL QUESTION SEASONAL FLUCTUATION OF PHLEBOTOMINAE (DIPTERA: PSYCHODIDAE) IN THREE DEPARTMENTS OF AN ENDEMIC AREA OF CUTANEOUS LEISHMANIASIS IN JUJUY PROVINCE. A PRELIMINARY REPORT Boumezzough A, Kahime K, Boussaa S & Ouanaimi F 142 Faculté des Sciences Semlalia, Université Cadi Ayyad, Marrakech, Morocco [email protected] Keywords: Leishmaniasis, Epidemiology, Vector, Sandfly, Morocco Remondegui CV1,2, Cabrera CHA2, Quintana MG2,3 1 Cátedra de Biología Celular. Facultad de Ciencias Agrarias – UNJu, 2 REDILA, 3 Instituto Nacional de Medicina Tropical – MSN, Instituto Superior de Entomología UNT-CONICET [email protected] Leishmaniasis is a parasitic disease a wide range of clinical symptoms which currently threaten 350 million persons in 88 countries. It caused by three Leishmania species, which are endemic, widespread and represent a public health problem in most countries in the Mediterranean basin. In Morocco, Leishmaniasis are endemic diseases constituting a major public health threat. Cutaneous leishmaniasis (CL) is caused by three clinically important Leishmania species (L. major, L. tropica, and sporadic CL cases due to L. infantum, flagellate protozoa of the family Trypanosomitidae). Visceral Leishmaniasis is also caused by L. infantum. This study presents a summarized analysis of its epidemiology in Moroccan context. And aims to determine the current leishmaniasis epidemiological situation in Morocco and the distribution of its different forms throughout the country. During the past 20 years, this disease has emerged as a major public health threat in Morocco. So, we gave a particular attention to vectorial status of Moroccan sandflies (Diptera: Psychodidae, Phlebotominae) in view of its major role in diseases spreading. It seems clear that the risk of spread of the disease is rising in Morocco. Ecological characteristic of proven and potential vectors should be regarded as good marker to anticipate leishmaniasis distribution in Morocco. Keywords: Phlebotominae, Jujuy, Argentina The endemic area of cutaneous leishmaniasis (CL) in the province of Jujuy includes eight departments. The number of recorded cases of leishmaniasis over the period 1990-2008 allowed ranking the risk areas to contract the disease, placing the area of "Ramal" as the greatest area of effective vector-parasite contact risk, concentrating almost 50% of cases in the Ledesma Department. In 2008 we started systemized studies to determine the presence of sandflies and were recorded specimens of Nyssomyia neivai (Pinto), Migonemyia migonei (Franca) and complex cortelezzii (Evandromyia cortezzii, Ev. sallesi). The aim of this work was to study the seasonal fluctuation of Phlebotominae species in three departments of the endemic area of the province of Jujuy, specially in sites with epidemiological antecedents. We conducted monthly sampling by 24 months from September 2011 in selected sites of Ledesma, Santa Barbara and San Pedro departments. The captures were made with CDC light traps by two consecutive nights. The specimens captured were clarified and preserved in eugenol until its identification to specific level. A total of 12013 sandflies were captured, a 51.5% corresponded to the Santa Barbara department, the 30.5% to Ledesma department and 18% to San Pedro department. In the three departments were recorded abundance peaks in autumn and spring. Preliminary results obtained in the present work show a increased abundance of sandflies in autumn and spring with a decrease in the abundance in summer and winter. The information resulting from this work on the diversity, abundance and distribution of Phlebotominae in different seasons and types of environments could be useful to detect effective measures aimed at reducing the population of these insects, avoiding the risk of outbreaks. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 143 ID 169-P Ecology & Epidemiology ID 170-P Ecology & Epidemiology SPATIAL ANALYSIS OF THE OCCURRENCE OF LEISHMANIASIS IN PARANÁ, BRAZIL, TO IDENTIFY AREAS OF RISK DENSITY DEPENDENCE IN HOST-VECTOR RELATIONSHIPS AND CONSEQUENCES FOR TRANSMISSION Alceu Bisetto Júnior, Maurício Bisetto, Sônia Maria Dotto Ampessam Erin Dilger, Orin Courtenay and Graham Medley Secretaria de Estado da Saúde do Paraná University of Warwick, UK. [email protected] [email protected] Keywords: Geoprocessing, American Cutaneous Leishmaniasis, Epidemiology Keywords: host choice, density dependence, Lutzomyia longipalpis This study aimed to compare the epidemiology and geographical distribution of American Cutaneous Leishmaniasis (ACL) in the state of Paraná in the period between 2008-2012 to spatial data altitude and climate of the state using geoprocessing tools to identify risk factors and areas for the occurrence of this disease. The occurrence of ACL is directly related to the introduction of susceptible man to places where the natural cycle of the disease is active, with the presence of the insect vector and animal reservoirs. For the survey of the historical series and epidemiological analysis based on state data from the Information System for Notifiable Diseases from the Ministry of Health of Brazil, with the record of notifications and investigations of ACL was used, covering the period 2008 - 2012. The calculation of epidemiological indicators was performed with the Tabwin/DATASUS program. To the geographical location of clinical cases, according to place of residence and infection, and the co-relationships of the geographical distribution of cases with the environmental characteristics of favorable climate and altitude of the occurrence Phlebotomines analysis of spatial disease distribution was performed, the map hypsometric the state of Paraná and according to Köppen climate classification, processed in a Geographic Information System - GIS, using the mapping function of Tabwin and georeferencing tools available. The results confirmed that regions with altitude level less than 600 meters from the sea and Cfa climate, according to Köppen , are at higher risk of contracting. Sandflies readily feed on a wide range of hosts, and the dynamics of preference between available host types are likely to be pivotal in determining the transmission of multi-host diseases, such as zoonotic visceral leishmaniasis. Sandfly host preferences dynamics are, however, poorly understood, with simple relationships conventionally assumed between host density, vector density and transmission. Using integrated modelling and fieldwork approaches, the preference of sandflies for key host types (dogs, humans and chickens) and associated force of infection was investigated over a range of vector and host densities. Following trap optimization and the minimisation of trapping bias, preferences for key host types over a range of vector densities were observed over a period of seasonal variation in Brazil. Preference fluctuations in response to changes in host density were also observed following experimental manipulation of chicken densities within sheds. These investigations indicate non-linearity in the relationship between sandfly preference upon different host types with vector and host density. Pheromone mediated aggregation behaviour in conjunction with host attractiveness may explain additional attractive effects at higher vector and host densities. Such aggregation in association with outdoor resting hosts may have far-reaching implications for understanding and manipulating transmission, and potentially indicate host density manipulation as a possible control measure. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 171-P Ecology & Epidemiology ID 172-P Ecology & Epidemiology GEOGRAPHICAL DISTRIBUTION OF NYSSOMYIA NEIVAI (PINTO) AND NYSSOMYIA INTERMEDIA (LUTZ & NEIVA) IN THE STATE OF SÃO PAULO, BRAZIL FIRST MEETING OF LUTZOMYIA LONGIPALPIS IN RURAL AREAS RELATED TO OUTBREAK OF CANINE VISCERAL LEISHMANIASIS IN THE NORTHWESTERN REGION OF SÃO PAULO, BRAZIL Claudio Casanovaa, Paloma Helena Fernandes Shimabukurob, Bruna Almeida Bressianic, Thaís Fernanda Lazaric, Eunice Aparecida Bianchi Galati d Eduardo Bergo, Lucimar Nascimento, Vera Camargo-Neves a Superintendência de Controle de Endemias, SUCEN, Mogi Guaçu-SP; b Fundação Oswaldo Cruz, Centro de Pesquisas René Rachou, FIOCRUZ, Belo Horizonte-MG, c FUNDAP/SUCEN, Mogi Guaçu-SP, d Faculdade de Saúde Pública, Universidade de São Paulo, USP, São Paulo-SP. 144 Superintendence Of Control of Endemic Diseases [email protected] Keywords: Lutzomyia longipalpis, epidemiology, entomology [email protected] Keywords: Nyssomyia intermedia, Nyssomyia neivai, Distribution, Cutaneous leishmaniasis vectors Records of the geographical distribution of both arthropod-borne diseases and the vectors involved in their transmission are essential for epidemiological surveillance as well as the accomplishment and formulation of control strategies. Nyssomyia neivai (Pinto) and Nyssomyia intermedia (Lutz & Neiva), which comprise the Nyssomyia intermedia s.l. complex, do not completely meet the criteria to be incriminated as vectors of American cutaneous leishmaniasis (ACL). However, their anthropophilic behavior and predominance in areas where autochthonous cases of ACL have been occurred allows to consider them as the main suspected vectors in areas where these two species are found in the Southeastern region of Brazil, particularly in São Paulo State. In the present study we provide updated information on the geographical distribution of N. neivai and N. intermedia in the state of São Paulo, where more than 6,000 human cases of ACL occurred in the last 10 years. A database with records for the presence of N. neivai and N. intermedia in the municipalities of São Paulo state was compiled, using more than 10 different sources between 1943 and 2013 (i.e. literature, examination of specimens deposited in entomological collections, entomological surveillance data). Our data clearly shows that there are three distinct distribution patterns for these vectors: an allopatric distribution of (i) N. intermedia in municipalities situated in the coastal region of the Paraíba river valley and of (ii) N. neivai in the vast area of the Atlantic Plateau region; and (iii) a sympatric distribution in municipalities situated in the high areas of Ribeira and Paraíba river valleys, where both species could be acting as vectors of ACL. Geographical distribution databases are useful to map epidemiological data and to build predictive models of disease. Lutzomyia longipalpis (Lutz & Neiva) is the main vector of Leishmania infantum chagasi (Cunha & Chagas), the etiological agent of visceral leishmaniasis in the State of São Paulo (SPS). This sandfly is well adapted to the peridomestic environment of the urban areas of the municipalities of West Paulista Plateau, Western Region of SPS. Since its first meeting in 1997, L. longipalpis has always been associated with the VL transmission only in urban environments in the state. This is the first reported finding of L. longipalpis in a rural area and the record of canine visceral leishmaniasis(CVL).In June and July 2013 were reported to the Superintendency of Control of Endemic Diseases 11 dogs with suspected CVL in a rural area, situated on the edge of the dam of the city of Pedregulho Estreito, at a place called Água Azul Ranch. The city of Pedregulho has 8118.4 km ² and is geographically located in the central part of the SPS and occupies 2.27% of the total area of the state. It is situated near the basins of the Paraná and Paraguai rivers and geographical coordinates are 20 ° 26'34 "South, 54 ° 38'47" West. According to Köppen's climate is Tropical Altitude, with dry winters and mild summers. He is presenting the rainy season is during the summer months (November to March). The maximum average annual temperature is 28.5 °C and minimum of 9.4°C in the winter months (July-August). The average annual rainfall is 1545 mm of rain. The entomological captures were performed onsite Agua Azul Ranch, where these dogs lived.In October and November 2013, considered one of the greatest periods of vector chance meeting. Five light traps bait (AIL) model modified CDC (Center for Disease Control) were exposed for three consecutive weeks, and installed at five different points peridomicile (next to the kennel, chicken house and the house owner, the porch and around the tank water) As a result 43 individuals were captured: 53.5% were male and 5 species of sandflies were identified: Nyssomyia whitmani (44.4%), Evandromyia lentivirus (22.2%), Lutzomyia longipalpis (18,5%), Evandomyia cortellezzi (7.4%) and Psychodopygus aragaoi (7.4%). Although most species caught were preserved in this environment both Lutzomyia longipalpis and Nyssomyia whitmani (the main vector of cutaneous leishmaniasis in ESP) were captured in greater abundance in peridometic environments and close to shelters for pets (kennel and chicken coop). Contrary to what occurs in urban areas of the municipal. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 173-P Ecology & Epidemiology ID 174-P Ecology & Epidemiology STUDY OF SAND FLY COMMUNITY IN TWO RURAL ENDEMIC AREAS FOR AMERICAN CUTANEOUS LEISHMANIASIS IN THE MUNICIPALITIES OF MOGI GUAÇU AND MOGI MIRIM, SÃO PAULO STATE, BRAZIL DNA DETECTION OF LEISHMANIA SP. IN FEMALE SAND FLIES (DIPTERA: PSYCHODIDAE) IN BELO HORIZONTE CITY AND SUMIDOURO STATE PARK, MINAS GERAIS – BRAZIL. 1 2 Fernanda Elisa Colla-Jacques , Claudio Casanova , Fredy Galvis Ovallos3 1 Departamento de Biologia Animal, Instituto de Biologia, Universidade Estadual de Campinas, Campinas-SP; 2 Superintendência de Controle de Endemias, Mogi GuaçuSP, 3Programa de Pós Graduação em Saúde Pública, Faculdade de Sáude Pública, Universidade de São Paulo, São Paulo-SP. [email protected] 145 Saraiva, L.; Rugani, J.M.N.; Lima, A.C.V.M.R.; Reis, A.S.; Pereira, A.A.S.; Gontijo, C.M.F.; Andrade Filho, J.D. Grupo de Estudos em Leishmanioses/Centro de Pesquisa René Rachou/FIOCRUZ, Belo Horizonte, MG. [email protected] Keywords: Belo Horizonte city, Leishmania infantum, Lutzomyia longipalpis Keywords: Ny. whitmani, vectors, Leishmaniasis Historically, a significant number of American cutaneous leishmaniasis (ACL) cases registered in São Paulo State originated from the Mogi Guaçu River Valley, and generally, the sandfly vectors are abundantly present in this area. Based on these informations, this work aimed to study the sandfly community in two rural endemic areas for ACL in the municipalities of Mogi Guaçu and Mogi Mirim, in the North-east region of São Paulo State, by analysing the spatial and time distribution of the species in four different environments. During the study period, from November/2010 to October/2011, CDC electric light traps were installed every 10 days in two study areas area from 18:00 to 06:00 hours. In each one of the study areas, four traps were set up: one in the wooded area, one at the woods' edge, one in the peridomicile associated with animals (chicken shed) and one in the peridomicile associated to humans (porch). A total of 45,527 sandflies were collected, comprised of 20 species. Of this total, 60% were Nyssomyia whitmani, followed by Migonemyia migonei (19%) and Expapillata firmatoi (7%). Among the studied environments, chicken sheds were the most productive ones, where 85% of all the specimens were collected. To understand the ecology of these species, several indices were used to describe the sandfly communities. The chosen indices were species richness, diversity, similarity and standardized species abundance. The identification and dominance of Ny. whitmani, Mg. migonei and Ex. firmatoi, suspected vector of ACL agents, represents an increased risk for new cases. Visceral Leishmaniasis (VL) have high incidence and mortality in Belo Horizonte city and its metropolitan region. The present study evaluated the detection of Leishmania sp DNA in female sand flies at two area of cerrado biome know as Brazilian Savannah, one place is an urban area in Venda Nova Sanitary District, in Belo Horizonte, and a wild area of Sumidouro State Park, an environmental protected area from 50 Km of Belo Horizonte. The samples were collected monthly, during three consecutive days, from August 2011 to August 2013. Eighteen traps were used in Venda Nova Sanitary District, and twelve in Sumidouro State Park. Sand flies were identified following the classification proposed by Galati 2003. Female sand flies were subjected to DNA extraction procedures and tested for Leishmania DNA using the ITS-1 molecular target. Positive samples were sequenced for Leishmania species identification. Twelve females of Lu. longipalpis species were identified with positive detection for ITS-1 in Venda Nova Sanitary District, one being identified as Leishmania braziliensis, four as Leishmania infantum, and the others as trypanosomatids. Four females were positive at Sumidouro State Park, one of cortelezzi Complex with positive detection for Le. braziliensis, one of Psathyromyia lutziana species, one of Sciopemyia sordelli species positive for Chritidia sp. and one specimen of Evandromyia sallesi positive for trypanosomatids. The results shows that the detection of sand flies infected with Leishmania sp. was higher in urban ambient, with predominance of Le. infantum, which indicates an increased risk of transmission in this area. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina ID 175-P Ecology & Epidemiology ID 176-P Control ASSOCIATION OF OCCURRENCE AND ABUNDANCE O F L U T Z O M I YA L O N G I PA L P I S W I T H ENVIRONMENTAL VARIABLES AT DIFFERENT SPATIAL SCALES: MICRO AND MACRO-SCALE A PUBLIC HEALTH APPROACH TO LEISHMANIASIS CONTROL Santini MS1,2; Utgés ME2,3; Salomón OD2,3,4 1 Centro Nacional de Diagnóstico e Investigaciones en Endemoepidemias-ANLIS-MSN, Argentina; 2 REDILA,Argentina; 3 Instituto Nacional de Medicina Tropical-MSN, Argentina; 4 Consejo Nacional de Investigaciones Científicas y Técnicas. Keywords: Lu. longipalpis, occurence and abundance, visceral leishmaniasis Lutzomyia longiplapis is the species responsible of transmitting Le. infantum (Syn. chagasi) in America. The objective of this study was to evaluate the environmental factors that determine the occurrence and abundance of Leishmania spp. vectors in urban settlements. The study was conductedon during February 2013 in Santo Tomé City, Corrientes, Argentina. Fifty-three households were selected within each 600 m2 square using the “worst scenario” criterion. Sandflies were captured with mini-CDC light traps, active for 3 consecutive rainless nights. Environmental variables were gathered at two spatial scales: micro and macro-scale. At each household (micro-scale), a set of 6 variables were gathered at the same time of the entomological sampling (Trees near the trap, Fruit trees, Plant pots, Dogs, Hens and Unused materials). Macro-scale variables where obtained from the GPS (Altitude), or from a SPOT-5 image with 10 m of spatial resolution (Distance to stream, Distance to city border, Proportion of Urban cover, High and Low density vegetation cover, and Bare Soil cover). To evaluate the effects on accumulated abundance of Lu. longipalpis, we constructed a set of 13 models to account for micro-, macro- and mixed-scale effects. Models were either GLM's with negative binomial distribution, or Hurdle models. The final model was selected by means of the AICc criterion. Parameter estimates and BCa intervals of the final model where calculated by bootstrap based on 1000 replications. We captured a total of 853 sandflies belonging to six species: Lu. longipalpis, Migonemyia migonei, Nyssomyia withmani, Bruptomyia spp., Ny. neivai and Evandromyia cortelezzii-sallesi. The 98.35% of the sandflies captured were Lu. longipalpis. In relation to environmental variables and according to the final hurdle model, only Distance to the border of the city and High to medium density vegetation cover ended to be positively associated with the occurrence of sandflies in the city (macro-scale variables). Trees around the trap, Distance to the stream and its cuadratic, were the variables positively associated with the abundance of sandflies (micro and macro-scale variables). These results suggest that presence and abundance of Lu. longipalpis could be explained by different factors depending on the spatial scale considered. In these work, presence and abundance of sandflies were defined by different spatial scales. 146 Waleed Al Salem1,2*, Badriah Al Otaibi2,3, Louise Kelly-Hope1,4, Karina Mondragon-Shem 1 , Salah Balgonaeem 2 , Maha Abdeladhim 5 , Mohammed Al Zahrani2, Amir Hassan3, Jesus Valenzuela5, Alvaro Acosta-Serrano1,6 1 Parasitology Department, LSTM, England, 2Saudi Ministry of Health, Riyadh, Saudi Arabia, 3International Public Health, LSTM, R England, 4Neglected Tropical Disease Centre, LSTM, England, 5 Vector Molecular Biology Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Maryland, USA, 6Vector Biology Department, LSTM, England [email protected] Keywords: Old world cutaneous leishmaniasis, Saudi Arabia, Al Ahsa Old World cutaneous leishmaniasis (CL) is one of the most prevalent vector borne diseases in the East Mediterranean Region (EMRO). A combination of factors is responsible for the sustained spread of CL throughout this region, including urbanization, irrigation, governmental sector integration, socio-economic factors, lack of health education, civil war and human migration. Civil war in several nations in the EMRO region, including in Afghanistan, Syria, Yemen, Iraq, Libya and Lebanon, and mass human displacement triggered by these conflicts are responsible for the current spread of CL to new foci. Since the beginning of the civil war in Syria approximately 2.7 million Syrians have become refugees in Turkey, Lebanon, Jordan, Iraq and Egypt. Moreover 4.1 million Syrians have been internally displaced within Syria. Another consequence of conflict is the lack of leishmaniasis control activities, which can further exacerbate CL spread. Uncontrolled urbanization and labour migration has led to the appearance of several new foci in Saudi Arabia, made worse by the lack of a health impact assessment for the new settlements. Settlements in Northern Israel have experienced CL outbreaks because of their proximity to leishmaniasis endemic areas. Irrigation may also create conditions favourable to the establishment of new CL foci, by changing the surrounding environment of areas, such as the Sidi Saad Dam in Tunis. Notably, foreign troops stationed in the EMRO region tend to be highly affected by CL due to a lack of immunity to the parasite, especially in CL endemic areas. In Iraq, an estimated 2500 overseas soldiers have been infected by CL, of whom 1283 soldiers presented in 2003 following their tour of duty. Also the International Security Assistance Force (ISAF) reported 200 cases of CL at its bases in Mazar-e Sharif, Afghanistan. New control measures, based on a systematic public health approach, are urgently required to minimize the impacts of conflict and mass migration on CL spread. This study was carried out in the Al Ahsa governorate, located in the eastern region of Saudi Arabia. Al Ahsa is a large oasis surrounded by three deserts, Al Dahna, Al Summan and Empty Quarter, and has an estimated population of 1,165,422 people, comprised of 80% Saudis and 20% non-Saudis. Regarding health infrastructure, Al Ahsa has 18 hospitals and 68 primary health care centres. There are also three leishmaniasis clinics offering diagnosis and treatment: Al Yahya for the norther. N° 26 (1) 2015 Boletín de la Sociedad Entomológica Argentina 147 Boletín de la Sociedad Entomológica Argentina N° 26 (1) 2015 ISOPS VIII Country 1 Argentina 2 Bolivia 3 Brazil 4 Canada 5 Colombia 6 Authors Orals 52 Posters 1 6 326 24 2 10 86 42 8 6 Czech Republic 17 6 5 7 Ecuador 10 1 1 8 Ethiopia 6 4 2 9 France 13 2 10 Gabon 1 11 Germany 3 12 Guatemala 1 13 India 13 4 1 14 Iran 13 1 1 15 Israel 9 4 2 16 Italy 29 3 1 17 Japan 4 1 18 Kazakhstan 6 19 Madagascar 2 20 Mexico 6 1 21 Morocco 5 5 22 Nepal 3 23 Panama 8 24 Paraguay 8 25 Peru 26 Portugal 9 27 Saudi Arabia 8 2 1 28 Spain 25 3 6 29 Sudan 4 1 30 Switzerland 2 31 Tunisia 47 6 4 32 Turkey 23 2 3 1 24 1 1 1 1 1 1 1 1 6 3 148