CAN THO UNIVERSITY COLLEGE OF AQUACULTURE AND FISHERIES COMPARING DIFFERENT METHODS TO DETERMINE DIGESTIBILITY OF SNAKEHEAD (Channa Striata) JUVENILE BY NGUYEN THI THU NGAN A thesis submitted in partial fulfillment of the requirements for the degree of Bachelor of Science in Aquaculture Can Tho, January 16, 2013 CAN THO UNIVERSITY COLLEGE OF AQUACULTURE AND FISHERIES COMPARING DIFFERENT METHODS TO DETERMINE DIGESTIBILITY OF SNAKEHEAD (Channa Striata) JUVENILE BY NGUYEN THI THU NGAN A thesis submitted in partial fulfillment of the requirements for the degree of Bachelor of Science in Aquaculture Supervisor Ass Prof Dr TRAN THI THANH HIEN Can Tho, January 16, 2013 i Acknowledgements I wish to express my deep appreciation and sincere gratitude to my advisor Ass.Prof Dr Tran Thi Thanh Hien for her constant guidance and also good advises in during the time I did these experiments and wrote thesis The second thing I want to send nice messages to Mss Ngo Minh Dung and Mss Tran Le Cam Tu who transfer to me all of cultural professional knowledge when I cultured snakehead in wet lab or analyzed in the nutritional laboratory Thanks to students of advanced aquaculture course 34, Aquaculture A1 course 35, and Fisheries management course 35 who was enthusiastic to help me finish this thesis Finally, thanks my family and also relatives make all of good things from finance to spirit help me strong during time I did it ii Abstract In this study, a series of experiment was done to compare different fecal-collection methods and digestibility of some ingredients of snakehead A reference diet was fed hours interval on 24 hours to find down what time was appropriate for collecting feces in the next experiment and defined dry matter and protein digestibility of snakehead by settling technique No significant effect between groups of digestibility through 24 hours and time space from eight and ten hours was appropriate for applying three different methods (settling, dissection, and stripping) Stripping was ineffective in this case Dissection method was also assessed Dry matter and protein digestibility of this method were lower than those on settling method Therefore, settling was chosen as an appropriate method to determine dry matter and protein digestibility of test ingredient mixing with a control diet following 30:70 on ratio Ingredients included fishmeal, soybean meal, meat bone meal and blood meal The results showed that the diet which was made from fish meal was the best protein digestibility percentage (93.63±0.62), and the lowest one was meat bone meal (88.93 ± 0.58%) Soybean meal and blood meal were similar, 91.27 ± 0.58%, and 91.23±0.93%, respectively In digestibility of ingredients, fish meal was also the best one with 85.75± 1.90% on value, following by soybean meal (69.73 ± 2.3%), and lowest one being meat bone meal (52.28 ± 1.73%) In conclusion, some ingredients with animal protein sources were digested better than plant origin, except meat bone meal iii Table of Contents CHAPTER I INTRODUCTION 1.1 Background 1.2 Research objectives 1.3 Research contents CHAPTER II LITERTURE REVIEW 2.1 Characteristics of snakehead 2.1.1 Classification 2.1.2 Biology characteristics 2.1.3 Nutrition characteristics 2.2 Study on on fecal collection and digestibility of some ingredients for fish CHAPTER III RESEARCH METHODOLOGY 3.1 Experimental site and period 3.2 Experimental fish 3.3 Experimental facilities 3.3.1 Facilities 3.3.2 Experimental feed 3.4 Digestible series of experiments 3.4.1 Experiment 1: Detection appropriate time for collecting feces 3.4.2 Experiment 2: Determination of appropriate fecal collection method for digestibility of snakehead 3.4.2 Experiment 3: Determination of digestibility of feed ingredients for snakehead 12 3.5 Sampling and data analysis methods 14 iv CHAPTER IV RESULTS AND DISCUSSION 16 4.1 Environmental conditions 16 4.2 Comparison of fecal quantities and ADC settlement periods 17 4.3 Comparing three fecal- collection methods 18 4.4 Diets digestibility 19 4.5 Ingredients digestibility 19 CHAPTER V CONCLUSION AND RECOMMENDATION 22 5.1 Conclusion 22 5.2 Recommendation 22 Reference list 22 Appendix 25 v List of tables Table 3.1: Chemical composition of experimental ingredients Table 3.2: Table for predicted fecal-collection time 11 Table 3.3 Pellet – feed from mixing reference diet and ingredients 13 Table 4.1: Environmental condition of experiment 16 Table 4.2: The ADCs for three methods 18 Table 4.3: Diet digestibility of reference and treatment 19 Table 4.4: Digestibility of some ingredients 20 vi List of figures Figure 3.1: Settling system Figure 3.2 Stripping method 10 Figure 3.3 The structure of an intestine 10 Figure 3.4 Pellet feed after mixing 12 Figure 4.1: Dry weight and ADCs of snakehead on two hours interval 17 vii List of abbreviations ADCs Apparent digestibility coefficients FM Fishmeal SBM Soybean meal BM Blood meal MBM Meat bone meal RD Reference diet AIA Acid insoluble ash viii % A’: % nutrition in feed (dry weight) %B’: % nutrition in feces (dry weight)  Digestibility in ingredients ADCingredients = (ADCT – 0.7 x ADCR)/ 0.3 ADCR: % control diet digestibility (R diet) ADCT: % control diet digestibility (T diet)  Nutrition apparent digestibility of ingredients ADC Nu-treat: apparent digestibility coefficient of nutritional treatment ADC Nu-ref: apparent digestibility coefficient of nutritional reference N diet-ref: Nutrition of referent diet (%) N ingredient: Nutrition of ingredient (%) c Data analysis The criteria of these experiments (digestibility, chemical component of feed and feces) were analyzed mean, standard deviation by Excel 2010 and SPSS 16.0 and comparing difference between means among treatments basing on one way ANOVA with DUNCAN test, significantly, p < 0.05 15 CHAPTER IV RESULTS AND DISCUSSION 4.1 Environmental conditions During the daytime, pH didn’t change much between morning and afternoon time, varying from 7.8 to 7.95 That results made a good condition for fish growth because the pH level in culture tank was good for culturing snakehead from 6.5 to 9.0 (Truong Quoc Phu, 2003) Table 4.1: Environmental condition of experiment Temperature (0C) Dissolved oxygen (ppm) pH Morning 28.32±0.61 4.28±0.21 7.80±0.22 Afternoon 29.76±0.92 5.09±0.51 7.95±0.17 Values are means of triplicate group and presented as mean ± SD The table showed that the temperature in the afternoon higher than in the morning, but it didn’t fluctuate highly with average value 28.32oC – 29.76oC When increasing in temperature, enzyme was excreted more and went up in metabolism; evacuation rate would be faster than normal Which temperature level was appropriate for fish, it depended on kind of species The value from 25oC to 320C was considered good condition, so that system was appropriate for fish growth That systems were applied circulate system so the amount of oxygen didn’t change much between morning and afternoon time, changing from 4.28 -5.09 ppm 16 4.2 Comparison of fecal quantities and ADC settlement periods Figure 4.1: Dry weight and ADCs of snakehead on two hours interval The column chart showed that the dry weight was determined the highest value at eighth hours for feces collected (4.11g) and the lowest at 24 hours (0.57g) The value of apparent digestibility coefficient of 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24 hours were respectively 70.15%, 72.07%, 72.75%, 73.26%, 71.91%, 72.07%, 71.99%, 72.22%, 72.45%, 70.93%, 71.26%, 70.41% However, the highest amounts of dry weight of feces were collected at 8hours and the apparent digestibility coefficient was not significant differences between groups of each separate hours interval Therefore, after hours, settling, dissection and stripping methods were applied to collect feces of fish A research conducted by Hien et al (2009) on Tra catfish (Pangasinodon hypophthalamus), twenty fish (84g) was cultured on tanks (170L) and fed with a reference diet mixing with 1% of chromic oxide The feces also collected by settlement every hours The result showed that the fecal quantities collected at 14 hours after feeding higher compared with the other time The value of ADCs of feces collected of each separate two hours interval was no significant differences This result showed that 17 the appropriate fecal collection time depended on kinds of species and chemical composition of the diet 4.3 Comparing three fecal- collection methods Although all of three methods which were settling, dissection, stripping applied to snakehead, stripping was recognized not as a feasible method because of lack of feces for analysis in spite of doing three times There was difficult to collected adequate feces because of the structure of snakehead’s gut However, the settling and dissection were applied with enough feces for analyzing chemical composition (table 4.2) Table 4.2: The ADCs for three methods Fecal-collection methods Settling Dissection Stripping Apparent digestibility coefficient Dry matter Protein a 70.81 ± 1.81 89.84± 0.96a 21.04± 1.29b 41.11±1.43b - Values are means of triplicate group and presented as mean ± SD Values in the same columns having the same superscript letters are not significantly different (p>0.05) The value of dry matter ADCs calculated from sample or feces collected using settlement were 70.81% which was highly significant difference, comparing with those collected using dissection method was 21.04% Digestibility coefficients for protein were significant higher settlement than dissection method, respectively, 89.84% and 41.11% The ADCs of the reference calculated from sample collected using dissection method was lower those using settlement because there was mixture enzymes, feces and indigestible feed in the fish’s gut Stripping was ineffective method with snakehead but with a search on Gulf of Mexico sturgeon (Acipenser oxyrinchus desotoi) of Jesus A Venero et al (2006), stripping and dissection were applied in this experiment to clarify which were best apparent digestibility coefficients (ADCs) of dry matter, energy and crude protein Hand stripping resulted in ADC values of 58%, 69%, and 85% and dissection was not better 54%, 64%, 18 and 78%, respectively Stripping was the best effective method because of value in ADC and not kills all of fish There had many researches which were about different methods on different species Maybe this method was the best with this species but not in another Therefore, settling method was considerably the best one in this case because it showed high result in dry matter and protein apparent digestibility coefficients and didn’t kill all of experimental fish 4.4 Diets digestibility Table 4.3: Diet digestibility of reference and treatment Name ADC dry matter (%) ADCprotein (%) 72.54 ± 1.47b 91.69±0.68b Fishmeal 76.5±1.65c 93.63±0.72c Soya bean meal 71.7±0.77b 91.27±0.72b Meat bone meal 66.46±1.55a 88.72±0.58a Blood meal 71.49±2.23b 91.23±0.93b Control diet Values are means of triplicate group and presented as mean ± SD Values in the same columns having the same superscript letters are not significantly different (p>0.05) The table showed that all of value about dry matter and crude protein apparent digestibility coefficient was significant (p0.05) Protein digestibility was calculated and compared with ingredients; meat bone meal (83.46%) was ingredient which had the lowest digestibility Moreover, this ingredient made from by-product cattle which there are high mineral and low methionine on chemical composition when processing feed The highest one was still fishmeal with 96.82% on value In this case, there had a changing a little bit between groups of two others Blood meal (90.69%) was higher than soybean meal (90.35%) but was not significant difference (p>0.05) on ADCs for protein of ingredients Because snakehead was carnivore fish, this kind of fish have the tendency digest animal protein source more 20 efficient than plan protein origin although both of ingredients was low essential amino acid (methionine) In another case, an experiment was conducted by S.M Hussain et al (2011) determining the apparent digestibility coefficient of Labeo rohita fingerlings with fifteen fish per tank and 12g on weight There were three ingredients (fishmeal, meat meal, and blood meal), following 70:30 ratio and using chromic oxide likely an indigestible marker The result showed that the crude protein digestibility of blood meal (71.9% ±1.98%) lower than fish meal (80.20 ±1.27%) and meat meal (79.92 ±2.4%) A research on tropical catfish (Mystus nemurus) was undertaken by M.S.Khan (1994) using a test ingredients and reference diet following 30:70 ratio Some common ingredients included on this research, namely, fishmeal, soybean meal, rice bran, copra meal, maize, and chicken viscera Dry matter and protein were digested fairly well on result The dry matter ADC for fishmeal was 77.88% while that for soybean meal was only 95.55%, similarly with protein digestibility for fishmeal 97.8% and soybean meal was just 86% 21 CHAPTER V CONCLUSIONs AND RECOMMENDATION 5.1 Conclusion After feeding hours, it was the suitable time for collecting feces on comparing three methods experiment Settling was the best method for research digestibility of snakehead (Channa striata) Fish could digest protein in feed as well as all range of ingredients, fish meal which was 93.63%, was the highest value and lowest one which was about 88.72%, was meat bone meal Dry matter and protein digestibility in ingredients which were really high, over 69% accept meat bone meal (52.28%) on dry matter and those for protein digestibility was over 90 %, also accept meat bone meal (83.46%) 5.2 Recommendation Research on different kinds of ingredient determined which sources was cheap cost, good digestibility, and availability Reference list Amirkolaie, A.K., S.A EI- Shafai, E-H Ending, J.W.Schrama and J.A.I Verreth 2005 Comparison of fecal collection method with high and low quality diets regarding digestibility and feces characteristics measurements in Nile tilapia Aquaculture Research, 36:578-145 Degani, G., Viola, S & Yehuda, Y (1997) Apparent digestibility coefficient of protein sources for carp, Cyprinus carpio L Aquaculture Research., 28, 23- 28 22 Duong Nhut Long.2003 Giáo trình nuôi thủy sản nước College of Aquaculture and fishery, Can Tho University Fagbenro and Davies, 2001 Use of soybean flour (dehulled, solvent-extracted soybean) as a fishmeal substitude in practical diets for Africa catfish, Clarias gariepnius( Burchell 1822): growth, feed utilization and digestibility Journal of Applied Ichthology, 17: p.64 Glencross, B.D., Hawkins, W.E., Evans, D., McCafferty, P., Dods, K., Maas, R &Sipsas, S (2005) Evaluation of the digestible value of lupin and soybean protein concentrates and isolates when fed to rainbow trout (Oncorhynchus mykiss) using either stripping or settlement fecal collection methods Aquaculture, 245, 211–220 Hemre, G.I., Ø Karlsen, A.M Jensen and G Rosenlund 2003 Digestibility of dry matter, protein, starch, and lipid by cod, Gadus morhua: comparison of sampling methods, Aquaculture, 225: 225-232 Hertrampf, J.W and F Piedad- Pascual 2000 Handbook on ingredients for aquaculture feeds Kluwer Academic Publicsher, Boston, London, 537pp http://bayeranimal.com.vn/doctor/technical/73-ky-thuat-nuoi-ca-loc-thuong-pham.html K Samantary, S.S Monhanty 1997 Interactions of dietary levels of protein and energy on fingerling snakehead (Channa striata) Aquaculture 156, from 141 to 147 Jesus A Veneroa, Richard D Milesb & Frank A Chapmana, 2006.Validation of a Fecal Collection Method for Determination of Apparent Digestibility Coefficients of Diets in Gulf of Mexico Sturgeon Aquaculture Nutrition, p.90-94 Khan, M.S (1994) Apparent digestibility coefficients for common feed ingredients in formulated diets for tropical catfish, Mysyus nerumus (Cuvier & Valenciennes) Aquatic Fisheries Management 25, 167-174 Kim,.J.D, S.M Tibbetts, J.E Milley and S.P.Lall 2006 Effect of the incorporation level of herring meal into test diet on apparent digestibility coefficients for protein and energy by juvenile haddock, Melanogrammus aelefinus L.Aquaculture 23 NRC (National Research Council) 1993 Nutrient Requirements of Fish National Academy Press, Washington, D.C., USA.69 pp Peng Li,.XC Wang, R.W.Hardy and D.M.Gatlin.2004 Nutritional value of fisheries bycatch and by-product meals in the diet of red drum (Sciaenops acellatus) Aquaculture,236: 485-496 Sales, J and P.J.Britz 2001 Evaluation of different markers to determine apparent nutrient digestibility coefficients of feed ingredients for South African abalone (Haliotis midae L.) Aquaculture, 202: 113- 129 S M Hussain, M Afzal, M Salim, A Javid, T A A Khichi, M Hussain and S A Raza 2011.Apparent digestibility of fish meal, blood meal and meat meal on Labeo rohita fingerlings The journal of animal and plant sciences.page 807-81 T.T.T.Hien, N.T.Phuong, T.L.C.Tu & B.Glencross (2009) Handbook of assessment of methods for the determination of digestibility of feed ingredient for Tra catfish (Pangasinodon hypothalamus) Tran Thi Thanh Hien, Nguyen Anh Tuan (2009) Dinh dưỡng thức ăn thủy sản College of Aquaculture and fishery, Can Tho University Umesh, N.R.,K Dathathri, M.C.Nandeesha, B Gangadhar and T.J Varghese 1994 Digestibility of dry matter and protein for Spirulina platensis by common carp (Cyprinusncarpio) with a note on time of fece collection in digestibility estimation In De Silva, S.S (ed) Fish nutrition Research in Asia Proceeding of the Fifth Asian Fish Nutrition Workshop Asian Fish Sos.Spec.Publ Manila, Philippines, Asian Fisheries Society, pp.81-84 Usmani, N., Jafri, A.K & Khan, M.A (2003) Nutrient digestibility studies in Heteropneustes fossilis (Bloch), Clarias batrachus (Linnaeus) and C garienpinus( Burchell) Aquaculture Research.,34,1247-1253 Vala1zquez, M and F.J.Martinez 2005 Design and testing of feces- collecting device for fish digestibility studies using demand or automatic feeding Aquaculture, 33:126-134 24 Vandenberg, G.W and J.D.L Noüe 2001 Apparent digestibility comparison in rainbow trout (Oncorhynchus mykiss) assessed using three methods of feces collection and three digestibility markers Aquaculture Nutrition, 7:237-245 Ward, D.A., C.G Carter and A.T Townsend 2005 The use of yttrium oxide and the effect of fecal-collecting timing for determining the apparent digestibility of minerals and trace elements in Atlantic salmon (Salmon salar, L) feeds Aquaculture nutrition, p49 Weathrup, R.N and K.J Mc Cracken 1998 Comparison of estimates of digestibility of two diets for rainbow trout(Oncorhynchus mykiss) ( Walbaurn), using two markers and two methods for feces collection Aquaculture research, p.527 Appendix Appendix 1: ADCs for dry matter and protein in diet Dry matter ADCs in diets Duncan Treat 25 N Subset for alpha = 0.05 ment Dry matter ADCs in diet Duncan Treat ment Subset for alpha = 0.05 N 2 3 66.4600 71.4900 71.6933 72.5400 66.4600 71.4900 3 71.6933 72.5400 Sig 76.5033 1.000 452 1.000 Means for groups in homogeneous subsets are displayed 76.503 Sig 1.000 452 1.000 Appendix 2: ADCs for dry matter and Means for groups in homogeneous subsets protein in ingredients are displayed ProteinADCs Duncan DrymatterADCs Duncan Treat ment Sig Subset for alpha = 0.05 N 3 52.2772 69.0302 69.7258 71.4900 85.7512 1.000 576 1.000 Means for groups in homogeneous subsets are displayed Trea tme nt N Sig Subset for alpha = 0.05 83.4623 90.3464 90.6892 91.2233 1.000 588 96.8191 1.000 Means for groups in homogeneous subsets are displayed Appendix 3: ADCs of feces collected hours interval Duncan 26 Treatment Subset for alpha = 0.05 N 3 70.0467 12 70.3800 70.3800 11 70.9433 70.9433 70.9433 10 71.2667 71.2667 71.2667 71.8900 71.8900 71.8900 71.8967 71.8967 71.8967 71.9967 71.9967 71.9967 72.0467 72.0467 72.0467 72.1067 72.1067 72.1067 72.4800 72.4800 3 72.7567 72.7567 Sig 73.2433 087 051 27 059 Appendix 4: chemical composition of experimental ingredients Treatments Moisture Protein Lipid FM 86.46 63.22 5.25 FM 85.33 63.47 4.29 FM 85.11 62.36 4.13 SBM 92.48 46.51 3.15 SBM 92.59 46.98 2.89 SBM 92.45 48.09 2.64 MBM 83.52 52.77 9.47 MBM 83.60 54.44 10.24 MBM 83.79 51.75 10.29 BM 93.94 89.51 0.34 BM 93.81 92.41 0.19 BM 93.39 92.60 0.53 28 Appendix 5: Chemical composition of feed Treatments Moisture Protein Lipid Cr2O3 HRA FM 82.05 51.4 5.89 0.24 2.59 FM 82.08 51.4 5.61 0.24 1.90 FM 81.80 51.4 5.32 0.24 2.48 SBM 86.21 46.5 4.38 0.26 4.48 SBM 85.73 46.5 4.54 0.26 2.02 SBM 85.89 46.5 4.56 0.26 2.02 MBM 85.71 41.7 6.60 0.76 3.08 MBM 85.79 41.7 5.62 0.76 4.27 MBM 85.89 41.7 5.68 0.76 3.85 BM 88.00 60.3 2.56 0.33 2.16 BM 87.77 60.3 2.56 0.33 2.16 BM 88.06 60.3 2.56 0.33 2.16 RD 87.32 44.2 5.40 1.00 2.36 RD 87.45 44.2 4.37 1.00 2.13 RD 88.07 44.2 5.36 1.00 2.52 29 [...]... on snakehead (Channa striata) juvenile 1.2 Research objectives Detecting the appropriate fecal collection to determine the digestibility of some ingredients of snakehead juvenile in order to determine the protein digestibility of this species 1.3 Research contents Comparing different fecal collected methods to determine digestibility of snakehead (stripping, dissection and settlement) Comparing digestibility. .. on Tra catfish, determine the digestibility of some ingredients was significant for research nutrition of fish The results of research could help to find down the material, which fish could digest well to improve the efficient of culturing of fish Therefore, there were many kinds of research about digestibility, especially, the digestibility of material in different fish According to Tran Thi Thanh... According to Rainboth (1996), snakehead belongs to: Order: Perciformers Family: Channaidae (Ophiocephalidae) Genus: Channa (Ophiocephalus) Species: Channa striata Some sneakhead species lived in Mekong Delta: - Snakehead murrel (Channa striata) - Dwarf snakehead (Channa gachua) - Giant sneakhead (Channa micropeltes) - Splendid snakehead (Channa lucius ) 2 2.1.2 Biology characteristics According to Duong... terrestrial animal According to Tran Thi Thanh Hien (2004), the characteristic of marker is: (1) the speed of movement like as nutrient, (2) indigestibility and solubility in water, (3) no affecting digestibility and absorption of nutrients Because the inert markers do not affect digestibility and absorption of nutrients, the rate of marker in feed and feces is the digestibility of feed Some markers are... dissection and settlement) Comparing digestibility of the different rich protein ingredients of snakehead 1 CHAPTER II LITERTURE REVIEW 2.1 Characteristics of snakehead 2.1.1 Classification Channidea is the family of snakehead, including 2 genuses: one genus is Channa, lived in Asia and other is Parachana, lived mostly in Africa There are 30 species of snakehead today in the world, 5 species distributed in... According to Tran Thi Thanh Hien (2004), the amount of fiber in feed could highly affect to the digestibility of feed of aquatic animal Fiber could increase the speed of feed through digestive system so it could increase amount of intake feed and made slow growth of aquatic animal It is relevant with a research of Umesh et al (1994) about digestibility of protein and dry matter from Spirulina platensis... the other time The value of ADCs of feces collected of each separate two hours interval was no significant differences This result showed that 17 the appropriate fecal collection time depended on kinds of species and chemical composition of the diet 4.3 Comparing three fecal- collection methods Although all of three methods which were settling, dissection, stripping applied to snakehead, stripping was... Publicsher, Boston, London, 537pp http://bayeranimal.com.vn/doctor/technical/73-ky-thuat-nuoi-ca-loc-thuong-pham.html K Samantary, S.S Monhanty 1997 Interactions of dietary levels of protein and energy on fingerling snakehead (Channa striata) Aquaculture 156, from 141 to 147 Jesus A Veneroa, Richard D Milesb & Frank A Chapmana, 2006.Validation of a Fecal Collection Method for Determination of Apparent Digestibility. .. contributed to establish feed formula for aquatic animals (Glencross et al 2007) However, researches showed that the digestibility of feed ingredients was influenced by fecal- collection methods (Weatherup and Mc Cracken 1998; Vandenberg and de la Noue 2001; Glencross et al 2005) Therefore, the study aimed to detect the appropriate fecal collection methods and determine the protein digestibility of different. ..  Nutrition apparent digestibility of ingredients ADC Nu-treat: apparent digestibility coefficient of nutritional treatment ADC Nu-ref: apparent digestibility coefficient of nutritional reference N diet-ref: Nutrition of referent diet (%) N ingredient: Nutrition of ingredient (%) c Data analysis The criteria of these experiments (digestibility, chemical component of feed and feces) were analyzed mean,