THE EFFECTS OF Terminalia catappa LEAVES EXTRACT ON THE
Transcription
THE EFFECTS OF Terminalia catappa LEAVES EXTRACT ON THE
THE EFFECTS OF Terminalia catappa LEAVES EXTRACT ON THE HAEMATOLOGICAL PROFILE OF ORNAMENTAL FISH Betta splendens Rudy Agung Nugroho Dewi Saraswati World Aquaculture 2015 May 29, 2015 Jeju-South Korea Faculty of Mathematics and Natural Sciences Mulawarman University East Borneo (Samarinda)-Indonesia Overview and Outline Background The Aim and objectives Research methods Results and discussion Summary References Background • • • Terminalia cattapa L. known as (Indian almond tree) Indian almond is a Combretaceus plant (tropical almond family), presenting throughout any region of Indonesia. The plant is a large tree, which can reach up to 30 m height with a thick broad trunk; the leaves cluster toward the end of the branches with glossy, obovate blades mostly 8-30 cm in length and turn red before turning brown and falling Leavesfolk Medicine and antibacteria in human (In Southest Asia) In aquaculture: Form Species and effect Leaves extract Watchariya P, Surapon W. eliminate Zoothamnium spp. Nontawit A. 2004 infection of black tiger post larva shrimp (Penaeus monodon)within 24 hours after exposure Leaves extract promote regeneration of fin tail of fancy carp (Poecilia reticulata) (Chansue, & Assawawongkasem, 2008) Aquaeus extract (Red and green leaves) Invitro Antibacteria (28 (Purivirojkul 2012) Penaeus monodon post larvae Survival rate and growth performance (Ikhwanuddin et al., 2014) Aquaeus water extract Researchers Isolates bacteria) • Dried leaves extract Betta splendens in vivo test antibacteria (Chansue, & Assawawongkasem, 2008) • Aqueous extract of dried leaves of almondBetta splendens weight gain and feed conversion; behavior of animals (Santos et al., 2013) • The Indian almond leaves have been claimed as a promoting substance for wound healing, especially for injured Betta Siamese fighting fish after fighting • Affect on breeding of Betta • To enhance water quality in betta culture However, there is a limited information on the effects of various concentration of Terminalia cattapa leaves on the haematological profiles The aims and objectives The study was conducted to examine: The effects of various concentration viz: 125; 250; 375; 500 ppm of liquid extract of Terminalia catappa leaves (ETC) on the haematological profiles: • Erythrocyte, • leukocyte, • lymphocyte, • hematocrit, • hemoglobin, • granulocyte, • serum protein and total protein level ornamental fish (Betta splendens). Research methods Terminalia cattapa extract The leaves were dried in laboratory at room temperature • Alkaloid Dragendorff's test • Phenolic Harborne, 1996 • Steroid and triterpenoid Liebermann-Burchad test • Flavonoid Harborne, (Chansue, 2008). Phytochemical test (Qualitative) 1996 • Saponin Forth test • Tannin Zohra et al, 2012 • Quinon Kala et al., 2012 To yield 50 mg/mL concentration, 1 Kg of dried leaves was minced and soaked in 20 L of distilled water for 3h at room temperature Fish test The extracted solution was filtered (Whatman paper No. 4). Pre-experimental • • 75 fishes (Male) initial weight ±0.53 g, from local fish market kept in Animal Physiology laboratory, Mulawarman University Acclimation three days Fed with frozen blood worm 0.01 g/fish twice a day 09.00 and 15.00 GMT. Experimental Control 3 tanks 5 fishes 125 ppm 3 tanks 5 fishes 250 ppm 3 tanks 5 fishes 6 L freshwater+bath supplement (Various ppm ETC) Water quality checked twice a week Fed with frozen blood worm 0.01 g twice a day 09.00 and 15.00 GMT Siphoned before feeding, every day 375 ppm 3 tanks 5 fishes 500 ppm 3 tanks 5 fishes 17 days Erythrocyte, leukocyte, lymphocyte, hematocrit, hemoglobin, granulocyte, serum protein and total protein level By using Mindray and Biolis 24i Results Table 2. Haematological values of ornamental fish (Betta splendens) after ETC trial Parameters Erythrocyte (106/µL) Leukocyte (103/µL) Lymphocyte (103/µL) Hematocrit (%) Haemoglobin (g/dL) Granulocyte (103/µL) Serum Protein (mg/dL) Total Protein (mg/dL) Groups trial (ppm) Control 125 250 375 500 0.053 ± 0.012a 0.063 ± 0.218a 0.067 ± 0.024a 0.03 ± 0.152a 0.21 ± 0.090b 0.86 ± 0.145a 1.23 ± 0.466a 1.06 ± 0.317a 0.6 ± 0.057a 5.1 ± 1.802b 0.55 ± 0.202a 0.83 ± 0.352a 0.73 ± 0.202a 0.36 ± 0.033a 3.1 ± 0.960b 0.3 ± 0.152ab 0.36 ± 0.185a 0.4 ± 0.208ab 0.13 ± 0.133a 1.4 ± 0.709b 0.9 ± 0.100a 0.43 ± 0.088a 0.46 ± 0.033a 0.36 ± 0.066a 1.13 ± 0.175a 0.15 ± 0.057a 0.15 ± 0.057a 0.25 ± 0.622a 0.1 ± 0.00 a 1.35 ± 0.519a 0.5 ± 0.100a 0.43 ± 0.333a 0.5 ± 0.057a 0.5 ± 0.166a 0.53 ± 0.033a 0.13 ± 0.033a 0.16 ± 0.033a 0.16 ± 0.033a 0.16 ± 0.033a 0.2 ± 0.000 a Data are given as mean ± SE (Standard Error). Different superscript (a, b) in the same row indicate significantly different means at P<0.05. Liquid extract of Terminalia catappa Leaves (ETC) was used as a source of extract test Discussion Saponin, Quinon antifungal and antimicrobia (Khristyana, 2005; Cowan, 1999) Phenol antioxidant activity free radical (Kinsella et al, 1993) Tannin water quality pH and ammonia (Masduki, 1999) Flavonoidincreasing antibody and stimulation on lymphocyte systemsecretion of cytokinin InterlLeucin (IL-2) (Sashidhara et al., 2007; Hefni, 2013). Flavonoid influences on hematocrit value (DeNoon, 2004) Summary • ETC contains useful phytochemicals such as Saponin, Quinon , Phenol Flavonoid and Tannin. • 500 ppm of ETC is beneficial to enhance the number of erythrocytes, leukocytes, lymphocytes and the percentage of hematocrit of betta fish. References Chansue, N. and Assawawongkasem, N. 2008. The in vitro Antibacterial Activity and Ornamental Fish Toxicity of the Water Extract of Indian Almond Leaves (Terminalia catappa Linn.).Chulalongkorn University, Bangkok, Thailand.KKU. Vet. J. Vol. 18 No. 1. DeNoon, D. 2004. A dark chocolate a day keeps the doctor away : daily dark chocolate good for the heart, loaded with flavonoids.WebMD Medical News. http://www.webmd. com/content/article/88/99702/htm Ikhwanuddin, Mhd, Julia HZ Moh, Manan Hidayah, Abu B. Noor-Hidayati, Nur MA AinaLyana, and Abu S. Nor Juneta. "Effect of Indian almond, Terminalia catappa leaves water extract on the survival rate and growth performance of black tiger shrimp, Penaeus monodon post larvae." AACL Bioflux 7, no. 2 (2014): 85-93. Purivirojkul W., 2012. Potential application of extracts from Indian almond (Terminalia catappa Linn.) leaves in Siamese fighting fish (Betta splendens Regan) culture. European PubMed Central (PMID:23885412). http://europepmc.org/abstract/med/23885412 Santos, D. M., E. L. Santos, A. P. L. de Souza, M. C. Temoteo, MC de A. Cavalcanti, F. C. B. da Silva, and E. da C. Pontes. "Use of aqueous extract of dried leaves of almond (Terminalia catappa) in the cultivation of betta splendens." PUBVET 7, no. 4 (2013). Correspondence: Rudy Agung Nugroho*) Dewi Saraswati Faculty of Mathematics and Natural Sciences Mulawarman University Samarinda, Borneo-Indonesia Email: *) [email protected] Additional Slides How to make leaves extract of Terminalia cattapa wash Cut and dry Maseration 3 days How to count blood profiles Mindray 0.01 mL blood + 0.1 mL EDTA Erythrocye Leukocyte Limphocyte Granulocyte Hematocrit How to count Protein Serum and Protein Total 0.01 mL Blood + 0.1 mL EDTA 1000 rpm 15 minutes spleen+ 2 mL NaCl Tokyo Boeki Machinery Ltd. Biolis 24i Premium 2. UJI FITOKIMIA + 1 drop of dragendorff Alkaloid test (Dragendorff test) + Residue Extract Acid + + 3-5 drops H2SO4 2M + Chloroform – amoniak 0,005 M filter + Filtrate Base Flavonoid test Shake; Positivered, yellow or purple Boil 5 minutes Extract Residue +100 mL water Filtrate + Phenolic test 0,05 mg Mg and 1 mL HCl + few drops of FeCl3 1% Positive red green or purple, or black Extract Saponin test Positive stable spume for 10 minutes with 1-3 cm shake for 10 s + Extract + 10 mL hot water Steroid and triterpenoid test Steroid Green or blue TriterpenoidRed or purple Unsolved in dietil eter Extract + + 5 mL dietil eter + CH3COOH glacial and H2SO4 glacial + Solved in dietil eter Quinones (Kala et al 2012) 1 mL of extract added with 1 mL of con. H2SO4 formation of red colour shows the presence of quinones. Tannins test (Zohra et al., 2012) 2 mL ETC + 2mL water added 1-2 drops of diluted ferric chloride solution A dark green or blue green coloration indicates the presence of tannins