LEVEL 1
Transcription
LEVEL 1
MONITORING OF CYANOBACTERIA BLOOMS IN FRANCE AND SOME OTHER EUROPEAN COUNTRIES (Germany, Portugal and United-Kingdom) Hélène DUCOBU CRITT Bio-Industries Midi-Pyrénées-CRT INSA 135, Avenue de Rangueil 31077 Toulouse Cedex 4 FRANCE Tel: +33 (0)5 61 25 35 57 Fax: +33 (0)5 61 55 94 00 E-mail: [email protected] • France is divided into 22 régions and 96 departments And 6 hydrographic basins Water monitoring concerns several structures which act at different administrative levels LEVEL 1: NATIONAL Ministry of the Environment (Ministry of Ecology and sustainable Development) Mission: Preservation and protection of the water resource Ministry of the Health (Ministry of Health, Family and handicapped People) Mission: Sanitary protection of the water users LEVEL 2: Hydrographic Basin Water Agencies* (6) LEVEL 3: REGIONAL DIREN* (regional direction of the environment) * Field work DRASS (regional direction of the sanitary and social affairs) LEVEL 4: DEPARTEMENTAL DDASS* (departmental direction of the social and sanitary affairs) MONITORING OF CYANOTOXINS. From awareness to reglementation 1999: Program EFFLOCYA initiated by the Ministry of the Environment and the Ministry of the Health. Collaboration with scientists (among which the Pasteur Institute). Step I: An inquiry sent to the Water Agencies, DDASS, Universities… to evaluate the occurrence of algae blooms in France. Simple questions such as: « Did you observe a change in lake’s colour? What was the colour (green, blue-green or red)? Did you notice dead animals? During which month of the year did the bloom occur? Did you make an observation under the microscope? … RESULTS: Low rate of reply (only 30 %) but it was a first step in the awareness process. Interesting results: The blooms occurred during Summer mainly in water reservoirs used for bathing or fishing. Most of the blooms were green in colour. Microcystis and Anabaena appeared to be the dominant cyanobacteria. Cases of mortality have been deplored among fishes and birds. Almost all the departments were concerned by the problem. Step II: Measuring Work of a small group of laboratories. Presence of toxins in several lakes. Lack of data in some regions of France (South). Lack of collaboration and communication between the laboratories. Almost everything occurred in Paris! ÎAll the scientists working on cyanobacteria decided to meet each others to exchange their experiences and to federate. 2001: Creation of the GIS (Group of Scientist Interest) CYANOBACTERIA. At the same time, the EFFLOCYA program did stop… Partners of the GIS: Research Institutes: INRA, CNRS; Pasteur Institute of Paris; Museum of History of Natural Sciences of Paris; Commissariat of Atomic Energy (CEA) French Agency for Sanitary and Alimentary Security (AFSSA); Universities; CRITT Bio-Industries Midi-Pyrénées... Main objectives: ÎResearch (determinism of cyanobacterial blooms; molecular tools); ÎMonitoring (standardization of the methods, development of new tools); ÎRisk assessment; ÎRemedial measures; ÎEvaluation of the socio-economic impacts; ÎIndustrial valorisation of cyanobacteria; ÎCommunication (interface between managers and scientists). REGLEMENTATION Brazil: First country to propose and implement a reglementation at a national level (Portaria n°1-469; 2000). World Health Organization: Recommandations for drinking water (1999) and bathing waters (2000). Guide Value: 1 µg/l MCYST LR European Community: Many directives concerning water quality but none about the control of cyanobacteria. But a standardization of the protocoles is starting at a European level. FRANCE: Order in Council n° 2001-1220 about water for human consumption (in application at the end of 2003) includes new parameters such as MCYST LR. Recommandation DGS/SD7a-N°2003-270 (June 2003, in application immediatly) about the control of cyanobacteria and toxins in bathing waters. WHO RECOMMANDATIONS Alert levels Drinking water Bathing water Cels cyanos/ml Vigilance Level Actions Alert 1 Level µg Chla Cyanos/l 200 2000 20 000 Alert 2 Level TOXINES 100 000 0.1 1 10 0.2-1 µg/l Guidance Level 1 2-10 µg/l Actions 50 Niveau guide 2 20-50 µg/l MONITORING OF TOXIC CYANOBACTERIA IN COLLABORATION WITH 5 WATER AGENCIES (2000-2003) Artois-Picardie Rhin-Meuse Loire-Bretagne Adour-Garonne Rhône-Méditerrannée-Corse AIMS OF THIS STUDY ? It was the very first time that the Water Agencies participated to a monitoring of toxic cyanobacteria • Complete the EFFLOCYA data. Especially data missing in the South. •Identify and list the water bodies which represent a risk and that should be monitored regularly. •To test and optimize the protocoles. •To think about the possibilities of a long-term monitoring (sampling frequency, with which actors, who is going to finance?…). •To make the water managers and water users aware of this environment and health problem. •To think about the way to communicate the results (to inform without causing panic). HOW DID WE START TO WORK? •January 2000: At this time, the 6 Water Agencies had received the EFFLOCYA inquiry en did have some data about the occurrence of cyanobacteria blooms in the water bodies belonging to each basin. •March 2000: I organized for the 6 Water Agencies a seminary about toxic cyanobacteria and made them a practical and financial proposal for a screening of toxic blooms in the 6 hydrographic basins during the Summer 2000. The Water Agencies were asked to finance the study totally. •May 2000: 3 Water Agencies joined immediately. I had to work hard to convince the 2 other ones. And the last one refused for financial reasons. Not possible to get a financing « Inter-Agence » because of administrative delay. So, each Water Agency did finance a study on its own territory and did chose its own sampling frequency (lack of homogeneity). •June 2000: Elaboration and distribution of the protocoles. I elaborated them together with Ingrid Chorus, in Germany who was used to work with Water Agencies. Some Water Agencies did ask for a training about sampling (we organized a one day training). We also sent to each Agency field and sample sheets and specific containers for samples. •Summer 2000: Sampling by Water Agencies and by Bi-Eau for the Water Agency Loire-Bretagne. Sending of frozen material (Chla and toxins) and non-frozen material (algal determination) to our laboratory. How the Water Agencies did chose the water bodies to sample? • « Cyanobacterial and eutrophication history » of the water body • We decided to focus on water bodies used for drinking water production, bathing and irrigation • The Agency Artois-Picardie choose some rivers and canals • Sampling frequency: 1 or 2 times a month during July, August and Septembrer (until November for the AG Agency) • Inquiry to the DDASS and collectivities RESULTS •52,6 % of the samples (170 in total) did contain potentially toxic cyanobacteria. •37 % of the samples did contain toxins (MCYST) •17 % of the samples did contain toxins at concentrations higher than 1 µg/l RESULTS (Summer 2000) Water Agency Chla µg/l (min; max) % samples with cyanobacteria potentially toxic RM 0.5; 265 41.9 23 6.4 166 - LB 7.9; 159000* 93 89 61.9 51.3 14106.7 RMC 2.2 ; 118.9 31.3 6 6 14 - AG 0; 74 70 64 10.4 10.7 - AP 0; 70 27 3.3 0 0.06 - * scum % toxic samples % samples with [tox] > 1 µg/l [Tox] max µg/l [Tox] max in a bloom µg/l EXAMPLES OF BLOOMS. How to identify them visually? Bloom of Anabaena. Niveau de risque modéré 4m Wind Accumulation Accumulation = 100 fois 4 cm Vent Accumulation = 1000 fois Lac : vue en plan Profil du lac Vent Anabaena accumulating along the dam under the wind action Green pellets of Microcystis Cyanobacteria mud Picture by Bi-Eau Accumulation of cyanobacteria along the shore Picture by Bi-Eau FOLLOWING UP 2001: Monitoring of the water bodies that reprensented a risk in 2000. We worked only with 2 Water Agencies (LB and AG). For the other ones, the toxins did not reprensent a priority (no obligation to make controls). 2002: The Water Agency LB did stop the monitoring and we decided to concentrate our work on our geographic region. New study with the Water Agency Adour-Garonne. Monitoring of all the toxins (all the microcystins, Anatoxin a, Anatoxin a(s), saxitoxins and LPS). 2003: With the new reglementation about MCYST LR, the Water Agencies consider that it is not their role anymore to do the monitoring. The managers and the owners of the water bodies should take it over. The Water Agencies also do not feel responsible for the communication about cyanobacteria as it is essentailly a health problem. However, they will have to integrate the parameters « cyanobacteria and MCYST » in the context of the monitoring of ecological quality of water bodies (SEQ Plan d’EAU). The way to integrate these new parameters is presently under discussion. SCREENING OF TOXIC BLOOMS IN 23 WATER BASINS OF THE ADOURGARONNE BASIN. Data 2000-2001-2002 Localization of the water bodies Mas Chaban Cognac Lavaud Tardoire Neuvic Moulin Pinard Bort-les-Orgues Couze St-Etienne-de-Cantalès Cabrerets Charpal Villeneuve/Lot Parentis Loubéjac Roucarié Fontbonne Rassisse St Ferréol Roques Choldocogagna Cammazes Drinking water Bathing water Some examples of the cyanobacteria we found Filaments Anabaena sp: microcystins, neurotoxins [anatoxin a, anatoxin a(s)] and LPS. Microcystins and neurotoxins Planktothrix agardhii Oscillatoria sp Aphanizomenon sp Saxitoxins Colonies: Gomphospaeria (non toxic?) and Microcystis Microcystis wesembergii Gomphosphaeria sp M. flos-aquae M. aeruginosa Cyanobacteria occurrence: Bilan 2000-2001-2002 Mas Chaban Cognac Lavaud Tardoire Neuvic Moulin Pinard Bort-les-Orgues Couze St-Etienne-de-Cantalès Cabrerets Charpal Villeneuve/Lot Parentis Loubéjac Roucarié Fontbonne Rassisse Legend St Ferréol Roques Choldocogagna Absence de cyanobactéries Présence de cyanobactéries (> 10 % du plancton) Cammazes Cyanobactéries (> 90 % plancton) Mas Chaban Cognac Lavaud Tardoire Microcystines occurrence 2000, 2001 et 2002 Neuvic Moulin Pinard Bort-les-Orgues Couze St-Etienne-de-Cantalès Cabrerets Charpal Villeneuve/Lot Parentis Loubéjac Roucarié Fontbonne Rassisse Légende St Ferréol Roques Choldocogagna Absence de cyanobactéries Présence de cyanobactéries sans toxines Cammazes Cyanobactéries et teneur en toxines < 1 µg/l Cyanobactéries et teneur en toxines > 1 µg/l Cyanobactéries et teneurs en toxines > 10 µg/l AEP Roucarié 14 14 12 12 10 2000 8 2001 6 2002 4 Alerte 1 2 concentration m icrocystines (µg/l) concentration m icrocystines (µg/l) Fontbonne 0 10 2000 8 2001 6 2002 4 Alerte 1 2 0 août septembre octobre août m ois septembre octobre m ois BAIGNADE Parentis 14 14 12 12 10 8 2000 Niveau 1 2001 6 2002 4 2 0 concentration en m icrocystines (µg/l) concentration en m icrocystines (µg/l) Lavaud 10 8 Niveau 1 2000 2001 6 2002 4 2 < 0.1 µg/l 0 août octobre m ois août octobre m ois Concentrations en microcystines totales d’août à octobre dans les 4 stations suivies de 2000 à 2002 PROFILS HPLC STANDARDS MICROCYSTINES LR RR C Max d’absorption: 239 nm HPLC profile of a lake used for bathing Bloom of Microcystis (different species). Presence of numerous microcystin variants (YR, 1023-RR, 980-LR + 1030-YR, identified by mass spectrometry). Use of molecular tools to detect the toxic potential M 1 2 3 4 5 6 M Résultats amplification par PCR de la région mcyA. M : marqueurs de taille ; 1 : CHO 10 ; 2 : MC 08 ; 3 : BLO 07 ; 4 : P08 ; 5 : PCC7806 ; 6 : H2O. The other toxins •No neurotoxins [Anatoxin a, anatoxin a(s) and saxitoxins ] found. So far, none neurotoxins found in France. LPS found (difficult to separate the bacterial LPS from the cyanobacterial ones). This Summer, high concentrations in a water body used as row water for drinking water production. No microcystins, no neurotoxins but LPS 31.8 ng/ml in row water. Lack of sanitary references. Réf: In Filand, fever outbreak after consumption of a tap water containing 40 ng/ml LPS which corresponded to LPS concentrations ranging from 200 à 1000 ng/ml in row water. Did we fulfill our objectives? • Complete the EFFLOCYA data. Especially data missing in the South. We have now data at a national level. •Identify and list the water bodies which represent a risk and that should be monitored regularly. The sites are identified and the regular monitoring has still to be done (the new reglementation will reinforce and give more power to our advisies) •To test and optimize the protocoles. •To think about the possibilities of a long-term monitoring (sampling frequency, with which actors, who is going to finance?…). •To make the water managers and water users aware of this environment and health problem. •To think about the way to communicate the results (to inform without causing panic). Most difficult part. The communication is not working very well! It should change with the new reglementation as it is specified that the public should be informed by panels in case of health risk. ANALYSES ASKED IN THE NEW REGLEMENTATION STEP I: Observation under the microscope: Présence or absence of cyanobactéria? If presence of cyanobacteria STEP II: Biomass: Chla and counting Î risk level according to the OMS recommandations In our lab, we add a step here: measurement of the toxic potention by PCR. If negative, steps III and IV should not be necessary. STEP III: Research of microcystins with PP2a test If test + STEP IV: confirmation and identification of microcystines with HPLC or HPLC-MS Monitoring in other European countries • Monitoring very active in the Land of Berlin • Straight collaboration between the Institute for Water, Air and Soil Hygiene (German Federal Environmental Agency which is the representative of WHO in Germany) and the numerous Water Agencies (more than one hundred). Sampling: samples concentrated on plankton net and row water • No reglementation but information of the public by press articles and panels at the lake’s shore with a message such as « If you go in the water until your knee’s level and you do not see your feet anymore, there is a risk of cyanobacteria bloom » Portugal • Straight collaboration between the Ministry for the Health, the Ministry for the Environment and the scientists • Monitoring at a national level. Before starting the monitoring, they spent one year standardizing the methods (sampling, analyses…) • Information of the public and the managers through a nice folder United-Kingdom • Information of the doctors, pharmacist and veterinaries