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EFFECTS OF MEDICATIONS ON THE SURVIVAL OF WILD CAUGHT SNAKEHEAD (Channa striatus) FRY DURING WEANING TO PELLETED FEED Presented by -Wahida Haque (st105569) AARM, SERD 1 Introduction 2 Problem Statement Mortality of Snakehead fry during weaning to artificial or pelleted feed Overall objective To develop a suitable weaning method by using feed combinations along with medication to ensure high survival rate Specific objectives To improve the survival by application of three medications during weaning To quantify the effect of medication on survival rate To evaluate the effectiveness of three medications used by some farmers during weaning To provide some preliminary scientific information about snakehead fry weaning Scopes Improvement of weaning method and survival rate Identification and Reduction of the factors responsible for mortality Determine suitable medication Limitations Limited experimental units and supporting literatures Supply of quality and same aged seed, Timing and unavailability of raw materials supply, Cost of feed and drug Maintaining water quality, Environmental fluctuation Microbiological infection and identification, Cannibalism ,stress Specific drug for specific pathogenic infestation Budget shortage, wastage of time, money and labour due to 3 dependency on others Language and communication Experiment-1 4 Materials and Methods Experimental fish Wild caught Snakehead fry about 1-2.5 inch size Age of about 1 month (Marimuthu, 2007) Collected from Suphanburi fishers Initially held in big hapa for acclimatization of 5 days Stocking density 65 fry/ hapa Experimental units Hapa of 1 m2 size, 3 earthen ponds of 200 m2 Water depth about 30-40 cm inside hapa Hapa covered with net to avoid predation and escaping Materials and Methods cont’d Experimental indicator Survival rate Experimental drugs/medicines Formalin bath- 37% formaldehyde solution ; dose as 35ppm for 30 min with continuous aeration Oxytetracycline- 25mg/kg of body weight/day in feed for 10 days Rotab- 0.1% of Piperazine citrate /kg of feed fed at a rate of 1% of body weight/day, twice in three weeks 5 Ref: FDA 1992, Boonyaratpalin,1985, Yolande,2005, Noga,2000 Materials and Methods cont’d Experimental feed Trash fish and frog feed (40% protein) in feed combination as 100: 00, 80:20, 60:40, 50:50, 40:60, 20:80 and 00:100 Each feed combination used for 3 days Feeding protocol Fed 3 times/day (7 am, 12.00 & 5 pm)until satiation (Wee 1982) Management Weekly water filling Dead fish removal Checking of hapa and covers 6 Materials and Methods 7 Materials and Methods 8 Materials and Methods cont’d Size Grading By 0.75 and 1 cm grading net at the beginning and after10 9 days Length and weight measurement Randomly initial and final length of 20 fry was measured Water quality measurement Temperature, DO, pH measured daily NH3 and NO2 measured weekly Estimation of survival rate SR%= (No of final harvest x100)/No of initial stocking Data analysis by -Microsoft Excel, SPSS 16.0 One way ANOVA and Tukey’s HSD Post Hoc Experimental design-1 CRD with 3 replicate of each treatment Duration 3 weeks Initial fry stock Formalin Formalin + antibiotic Antibiotic Rotab No drug (control) Formalin + antibiotic + Rotab 10 Formalin + Rotab Rotab + antibiotic Experimental hapa Setup Chemicals/drugs Formalin (T1) Antibiotic (T2) Rotab (T3) Formalin + Antibiotic (T4) Formalin + Rotab (T5) Rotab + Antibiotic (T6) Formalin + Antibiotic + Rotab (T7) Control –no drug (T8) 11 All with same feed combination All size graded Pond-1 Pond-2 Pond-3 R1 R2 R3 1(8) 1(6) 1(2) 2(6) 2(8) 2(1) 3(7) 3(5) 3(3) 4(4) 4(1) 4(4) 5(1) 5(2) 5(7) 6(2) 6(4) 6(5) 7(3) 7(7) 7(8) 8(5) 8(3) 8(6) Inside parenthesis is the Position of hapa and outside the hapa number. Hapa= 24+24=48, grading in 10 days Results: Graph showing the survival rate of eight treatments in experiment 1 Survival rate% 100 c 80 b 60 40 bc bc c b a a 20 0 F A R F+A F+R R+A F+A+R C Treatments Survival rate of T1,T2, T3, T4,T5,T6, T7 and (T8) are 43.59±2.35, 61.03±3.88,71.28±6.41, 59.49±3.88,73.84±4.07, 84.62±10.09, 80.00±4.62,42.56±6.41 respectively 12 The bars with different superscript are significantly different (P<0.05) Water Quality parameters in experiment 1 Parameters Pond-1 Pond-2 Pond-3 Temperature C 28.02 ± 0.64a 27.95 ± 0.63a 27.69 ± 0.81a DO mg/l 4.65 ± 0.64a 5.51 ± 0.85b 5.57 ± 0.73b pH 7.76 ± 0.10ab 7.85 ± 0.18a 7.71 ± 0.20b NH3mg/l 0.04 ± 0.01a 0.12 ± 0.07a 0.06 ± 0.03a NO2 mg/l 0.02 ± 00ab 0.04 ± 0.01a 0.01 ± 0.00b Values with same superscript in a same row are not significantly 13 different (P<0.05) Experiment - 2 Materials and Methods is same as experiment 1 except Stocking density- 58 fry/ hapa Experimental drugs- only Oxytetracycline and Rotab Parasites count : Gross examination of the intestine by wet mount microscopy for the presence of Nematodes, Cestodes and Acanthocephalans (Kabata,1985). Sample size 25 fish/group (randomly selected) assuming parasitic carrier prevalence more than 10% (Kabata, 1985). The parasites were identified as Kabata (1985), Ribelin (1975), Roberts (1978 and 1974). 14 Experimental design-2 CRD with 3 replicate of each treatment Duration 3 weeks Initial fry stock Antibiotic No drug (control) Rotab Rotab + Antibiotic 15 Experimental hapa setup in four treatments Chemicals/drugs Feed combination All size graded 16 Pond-1 Pond-2 Pond-3 R1 R2 R3 Rotab (T1) 1(3) 1(4) 1(1) Antibiotic(T2) 2(2) 2(2) 2(2) Rotab + Antibiotic(T3) 3(1) 3(1) 3(4) Control –no drug(T4) 4(4) 4(3) 4(3) Inside parenthesis is the Position of hapa and outside the hapa number. Hapa=12+12=24, grading in 10 days Results: Survival rate % 100 80 Graph showing the survival rate of four treatments in experiment 2 a a b c 60 40 20 0 R (T1) A (T2) R+A (T3) C (T4) Treatments Survival rate of Rotab (T1), Antibiotic (T2), Rotab+ Antibiotic (T3) 17 and Control -No drug (T4) are 93.11± 2.98,84.48 ±1.73,94.25 ± 2.63 and 72.99 ± 6.06 respectively with stocking density 58 fry/hapa The bars with different superscript are significantly different (P<0.05) Water Quality parameters in experiment 2 Parameters Pond-1 Pond-2 Pond-3 Temperature C 33.17±1.33a 32.77±1.29a 32.38±1.29a DO mg/L 5.15±1.60a 5.28±1.34a 4.31±1.53a pH 7.24±0.47a 7.23±0.35a 7.31±0.26a NH3mg/L 0.05±0.01a 0.15±0.08a 0.06±0.01a NO2 mg/L 0.03±0.01a 0.03±0.01a 0.03±0.01a No parameter showed significant difference among ponds ; Values 18 with same superscript in a same row are not significantly different (P<0.05) Parasites density Mean number of parasite per fish 6 5 a a a b 4 c 3 b 2 1 0 a a Initial Stock Control Sample group 19 a Rotab treated Nematode Cestode Acanthocephala Healthy , infected dead & moribund fishes 20 Parasites in experimental fishes 21 Discussions Carnivore, surface feeder (Sampath, 1979) Cannibalism obvious, (Qin, 1996) reduced by size grading and satiation feeding Fed as protein requirement,35-40% (Samantaray et al.,1997) and at 5% of body wt./day for juveniles for optimum growth (Qin, 1996,1997) Stocking density was appropriate (75-400/m2 Wee, 1982, Aonanong, 2008) Fry survival low, 7-24%, if stocked immediately in pond after collection from wild then survival is only 13-15% (Boonyarapatlin, 1985) 22 Discussions 3 major groups of pathogens found in snakehead Bacteria: Aeromonas sp. Flavobacterium sp., Pseudomonas sp., Haemophilus sp. External parasites: Protozoa and Fungi, monogeneans Internal parasites: Helminthes- cestodes, nematodes and Acanthocephalus (Ref: Boonyaratpalin ,1985, Areerat ,1981, Nash ,1988, Chinabut, 1983, Kaewpaitoon, 1992) Nematodes (Camallanus spp. and Spinitectus spp), cestodes (Senga spp) and acanthocephalans (Acanthocephalus spp., Gorgorhynchus spp. and Pallisentis spp.) have been found in our experimental fishes The average nematode and acanthocephalan number of initial stock (4.52/fish &5.32/fish) didn’t show significant difference with control treatment (4.24/fish and 4.24/fish) but showed significant reduction in Rotab treated (1.84/fish and 2.88/fish) fish sample. 23 Discussions In first experiment, Higher survival in Rotab +OTC combination treatment Rotab alone and in combination with OTC showed better survival than formalin and control treatments Formalin found non-effective to improve SR as formalin treatment had no variation with control indicate no significant presence of external pathogen The potentiality of endoparasites and bacterial pathogens assumed Second experiment was conducted to confirm the previous result Higher survival in Rotab + OTC treatment. Rotab alone and OTC+ Rotab combination has significant variation with control No significant difference (P > 0.05) of survival rate Rotab (93.11 ± 2.98) and combination of Rotab + OTC (94.25 ± 2.63) treatments. This confirms that Rotab is effective and the use of Rotab as a feed additive is sufficient to maintain relatively high survival during the weaning 24 Discussions In china, OTC used in feed as 50-80 mg/kg body wt./day for 4-6 days as “non-pollution fishery drug” and not different among species (Wang,2004) OTC depletion occur in fish tissue (Meinertz,2001) though it may present for 4-6 weeks in tissues after administration (Heijden 1995) Microbial control has greater influence on the survival and growth of fish larvae during the transitional feeding stage and addition of OTC in feed improves survival (Battaglene, 2006). OTC in food improved survival during weaning and caused less grey gut disease in striped trumpeter fish (Battaglene, 2006). OTC is effective and recommended against some major bacterial pathogens of snakehead (FDA,1992, Boonyaratpalin, 1985, Chinabut,1990 etc) 25 Discussions Some Thai farmers use Rotab (Aonanong, 2008 ) and OTC during weaning to reduce mortality but no scientific literature available Rotab is Anthelmintic for veterinary and key ingredient is Piperazine which has efficacy to reduce internal helminths and acanthocephalans (Santamarina,1991, Tojo,1998) Piperazine is effective and recommended against nematodes also effective as antiprotozoal in fish (Quintela,2003, Parama,2004), so protozoan also might be treated with Rotab in our study Non-toxic to aquatic organisms and environment, no potential for bioaccumulation, biomagnification or secondary poisoning (Noga,2000, Fairfield,2000,Youlande,2005) (CSTEE, 2004, EU report,2005) 26 Discussions Low quality trash fish increase disease incidence (Wee,1982),pelleted feed is better due to ensured quality (Kaewpaitoon, 1992, Aononong,2008,Wee,1982) Potential pathogen source are trash fish as fishes fed marine trash fish found to be infected with internal parasites (Chinabut,1990, Limsuwan, 1983, Kaewpaitoon, 1992) and wild source before collection Body symptoms indicate intestinal disease and infestations (Chinabut,1990, Areerat,1981,Limsuwan, 1983, Kaewpaitoon, 1992) In general,2nd experiment showed the better survival -better parents and source with better immunity, season The 1st experiment conducted in cooler months but 2nd experiment in 27 summer. Colder temperatures increase disease susceptibility and reduce growth (Kaewpaitoon,1992, Vivekanandan,1977) Water quality didn’t show much fluctuation in each experimental period and was within tolerable range (Ng, 1990, Qin, 1997, Courtnenay,2004, Rath, 2000) and it doesn’t cause heavy mortality in snakehead but may affect the growth and production (Wee,1982) Conclusions Rotab alone or in combination with OTC found to be effective and 28 showed higher survival rate but no variation between them Fishes were infected in wild before collection. Nematode and acanthocephalan density showed reduction in Rotab treated sample Formalin treatment found to be ineffective to improve survival Both Rotab and OTC used as approved dose so no human health and environment risk Rotab used by some farmers is effective and the use of Rotab as a feed additive is sufficient to maintain relatively high survival during the weaning phase of wild caught snakehead fry to artificial dry feeds. Finally can be said that, medications with Rotab and OTC together along with the size grading can improve the survival rate of snakehead fry during weaning to pelleted feed and these drugs, used by some farmers is found to be effective to achieve better survival rate. Recommendations Genetic study of fry Study of the source of fry Determine and Use of pathogen species specific drugs and dose The quality and shelf life assessment of the fed trash fish The microbiological study of the fed trash fish Impact assessment of the use of these drugs on other spp Effect of these drugs on the physiology of the fishes need to be assessed 29 Few References Battaglene, S.C., Morehead, D.T., Cobcroft, J.M., Nichols, P.D., Brown, M.R., Carson, 30 J. (2006).Combined effects of feeding enriched rotifers and antibiotic addition on performance of striped trumpeter (Latris lineata) larvae. Aquaculture 251: 456– 471. Boonyaratpalin, M., McCoy, E. W., Chittapalapong, T, (1985) Snakehead Culture and its Socio-Economics in Thailand, National Inland Fisheries Institute Kasetsart University and NACA Head Office, Bangkok, Thailand Kaewpitoon, K. (1992). Utilization of septage-raised tilapia (Oreochromis niloticus) as feed for snakehead (Channa striata). AIT Dissertation. 212pp. Marimuthu, K and Haniffa, M. A., (2007). Embryonic and larval development of the Striped Snakehead Channa striatus. Taiwania, 52(1):84-92. Nash, G., R.J. Roberts, S. Chinabut, S. Areerat and C. Limsuwan. 1988. Emaciation of pond-cultured snakehead, Channa striatus (Fowler). J. Fish. Dis., 11: 215-224. Noga, E.J (2000). Fish Disease : Diagnosis and Treatment. Wiley –Balckwell publishers. Book (367 pages). 2nd edition, page 292. Qin, J., Fast, A. W. (1996). Size and feed dependent cannibalism with juvenile snakehead Channa striatus. Aquaculture 144: 313-320 Samantaray, K., Mohanty, S.S. (1997) Interactions of dietary levels of protein and energy on fingerling snakehead, Channa striatus. Aquaculture. 156: 241-249 Wee, K.L (1982). The biology and culture of snakeheads. In: J.F. Muir and R.J. Roberts (edit) Recent advances in aquaculture. Westview press, Boulder Co. pp180-211. Thank You 31