effect of feed supplementation formulated from ...

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EFFECT OF FEED SUPPLEMENTATION FORMULATED FROM DIFFERENT PLANT PROTEINS SOURCES ON THE GROWTH PERFORMANCE OF CIRRHINUS MRIGALA AND CYPRINUS CARPIO Shahid Mahboob Department of Zoology, College of Science, King Saud University, P.O. Box 2455, Riyadh-11451, Saudi Arabia Email: [email protected] ABSTRACT Freshwater fish culture practices warranted a demand for efficient, balanced and economical fish feed. The aim of this study was to evaluate the effect diet composed of plant proteins as a replacement for fish meal for Cirrhinus mrigala, and Cyprinus carpio and to use the findings to prepare a cost effective and efficient diet for freshwater fish species. The plant protein feed (35 % crude protein) was composed of five different ingredients. The feed was fed at the rate of 0.15 g nitrogen per 100 g of wet fish weight daily for one year. Fish were fed twice per day by dusting and control pond devoid of any feed. C. mrigala and C. carpio performed in a curvilinear manner at different water temperatures. The weight gain in C. carpio was better with the feed supplementation compared to C. mrigala. In C. mrigala and C. carpio weight gain at the completion of the study, expressed as a percentage of the weight over control pond computed as 273.25 and 329.40 percent, respectively with the effect of feed prepared from plant sources by replacing the fish meal. KEYWORDS: Feed; Plant protein; condition factor; Growth; Weight INTRODUCTION Fish is today considered as the most promising inexpensive alternative source of animal protein. Fish offers a relatively high amount of essential amino acids, minerals and fatty acids as compared to livestock (Huisman, 1989). The share in the total world food production of animal origin is some 12%, the production ratio of fish to meat being 0.6 (Huisman, 1989). The role of fish in Pakistan, like many developing countries, is more important than this ratio suggests. Since it is more and more realized that capture fishery resources are not unlimited, emphasis has been given to the enhancement of intensive fish farming in order to bridge the increasing gap between demand for and supply of fish. Fortunately, Pakistan is endowed with vast expanse of inland fisheries resources which at present are not being adequately exploited due to lack of research and other technical facilities for the commercial fish farmer. The demand for seafood continues to increase, and aquaculture production has filled the shortfall associated with static wild fish landings (Fish Stat Plus, 2010). In fact, in 2012

aquaculture production is expected to exceed capture fisheries as a source of finfish products for consumption (FAO, 2010). Aquaculture production is expected to increase further and this will require higher production of aqua feeds. The inclusion of plant-protein sources in aqua feeds has increased due to the limited amount and increasing cost of fishmeal available for production of animal feeds (e.g., Gatlin et al., 2007; Glencross et al., 2005; Naylor et al., 2009). One of the greatest challenges for the aquafeed industry is to reduce fishmeal levels in feed further and increase the amount of plant protein and ingredient diversity in the diets of carnivorous fishes. Among commonly used feed ingredients, fish meal is considered to be the best ingredients, due to its compatibility with the protein requirement of fish (Alam et al., 1996). Many different plants-protein sources have been examined including plant-protein meals and plantprotein concentrates (Lim et al., 2008). According to Bhosale et al. (2010) increased use of plant protein supplements in fish feed can reduce the cost of fish meal. The research has focused on utilizing less expensive and readily available resources to replace fish meal, without reducing the nutritional quality of feed (Mahboob and Sheri, 1998). The purpose of this study was to test plant protein ingredients as replacements for fishmeal in diets for Cirrhinus mrigala, and Cyprinus carpio and to use these findings to develop and practice a feed formulated from plant sources to enhance production of these fish species. MATERIALS AND METHODS Four newly dug earthen fish ponds of dimensions 15m X 8m X 2.5m (length X width X depth) were used for this investigation. All the ponds were sundried for a period of fifteen days. In order to ensure disinfection of these ponds, liming was done with CaO at the rate of 2.40 kg/pond (200 kg/ha as suggested by Hora and Pillay, 1962) with dusting method. The inlets of all these ponds were properly fixed with screen to avoid the entry of any intruder into or exit of fish from the ponds. All the ponds were filled with unchlorinated tube well water up to the level of 2.0 m and this level was maintained throughout the experimental period so as to have a pond volume of 240.00 m3 for fish cultivation. A total of 84 four months old fingerlings of Cirrhinus mrigala (weight 1.68±0.02 g; fork length 51.68±0.06 mm) and Cyprinus carpio (weight

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WWW.AFINIDAD.ORG 18.49±0.10 g; total length 81.32±0.0.07 mm was stocked in each of the ponds with the stocking density of 2.87 m3/fish (Javed, 1988). The interspecies ratios were 50:50. Preparation of feed: The plant protein feed (35 % crude protein) was formulated five different ingredients, sesamean oil cake (SOC), maize gluten meal (MGM), cottonseed meal (decardicated; CDM), wheat bran (WB), rice polish (RP; Table 1). SOC, MGM, CDM, WB and RP (Table 1) were combined and vitamin and mineral mix to produce fish feed that contained digestible protein levels similar to menhaden fishmeal (Select grade). The rationale behind the protein-blend approach was that in commercial feed formulations, reducing fishmeal levels is best accomplished by combining several alternate protein sources to approximate the amino acid profile of fishmeal. Supplementing protein blends with amino acids further improve the nutritional profiles of blends in comparison with that of fishmeal (Cheng et al., 2003). Minerals shown to be important when feeding fishmeal-free diets were supplemented to each of the three blends (Barrows et al., 2010). Fish feed was prepared using commercial manufacturing technology. The feed was stored in plastic lined paper bags at room temperature until fed. The field trial was conducted with three replicates and one control pond. The feed was fed at the rate of 0.15 g nitrogen per 100 g of wet fish weight daily for one year. Each pond contained 84 fish and feed fed to three replicate ponds, arranged in a completely randomized design. Fish were fed twice a day by dusting. However, control pond remained without any additives. Table 1: Composition of feed constituent (35 C.P) used to prepare test diet in Cirrhinus mrigala and Cyprinus carpio feed Ingredients Seasamean oil cake Maize gluten meal (30 %) Cotton seed meal (decardicated) Wheat bran Rice polish Vitamins and mineral mix Crude protein

Percentage 32.0 20.0 40.0 3.5 3.5 1.0 35.o

Growth Studies The cultured fish stock was sampled, randomly, with three repeats on every 16th day (designated as fortnight) by using nylon drag nets from each of the ponds during the 12-months trial. The

morphometric characteristics of fish viz., Wet body weight, wet fork length and wet total length were measured and recorded to monitor growth back into their respective ponds. The sample size for each fish species remained 7 as determined by the following formula: n = t2s2/d2 t = Value of t from the normal probability table against α = 0.05 s2 = The variance among units in the population from previous work (Javed, 1988) d = Desired margin of error in the estimate (one percent) Statistical analysis The data were subjected to statistical analysis by using Minitab software. The differences among treatments were tested using ANOVA followed by the DMR test. RESULTS AND DISCUSSIONS The final average weight after trial 12 moths in C .mrigala and C. capio in treated and control ponds were recorded as 482.57± 2.87, 156.84 g ± 1.22 and 1566.77 ± 5.44 and 475.63 g ± 4.28, respectively (Table 2). In C. mrigala a gradual increase in weight was observed from the start of the experiment to the end of the 24th fortnight (December). The controlled fish exhibited the maximum increase in weight during the 16th fortnight with the increment of 17.80 g, closely followed by the increment of 14.36 g during 17th fortnight. C. carpio showed a gradual increase in weight from the start of the experiment upto 8th fortnight and showed a maximum increase of 194.48 g during 8th fortnight, closely followed by the 7th during which fish showed an increase of 140.57 g. After 8th fortnight gradual decrease in growth rate of fish was recorded upto 15th fortnight. After 15th fortnight, once again abrupt change in the growth rate of fish observed during 16th and 17thfortnights with the increments of 76.42 and 54.78 g, respectively (Table 2). It was observed that C. mrigala and C. carpio performed in a curvilinear manner at different water temperatures. C. carpio responded more efficiently to the feed compared to C. mrigala. In C. mrigala and C. carpio weight gain at the completion of the study, expressed as a percentage of the weight over control pond computed as 273.25 and 329.40 percent, respectively with the effect of feed prepared from plant sources by replacing the fish meal.

Table 2: Fortnightly average increase in body weight (g) of Cirrhinus mrigala and and feed supplemented pond

Fortnight

Cirrhinus mrigala Feed Supplemented Control pond pond Av. Wt Av. Wt Inc. (g) Inc. (g) (g) (g)

Cyprinus carpio in control

Cyprinus carpio Feed supplemented Control pond Av. Wt Av. Wt Inc. (g) Inc. (g) (g) (g)

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WWW.AFINIDAD.ORG 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

2.13 3.32 4.85 7.84 12.16 17.38 24.87 33.46 44.65 65.89 92.53 125.36 162.79 203.97 241.46 275.12 305.59 333.65 356.18 376.49 395.33 410.95 421.28 428.57

0.45 1.19 1.53 2.99 4.32 5.22 7.49 8.59 11.19 21.24 26.64 32.83 37.43 41.18 37.49 33.66 30.47 28.06 22.53 20.31 18.84 15.62 10.33 7.29

1.84 2.51 3.39 4.43 4.92 7.65 9.72 12.89 16.08 21.74 28.26 37.87 48.21 60.81 73.06 90.86 105.22 118.03 129.10 138.68 145.92 150.39 154.16 156.84

0.16 0.67 0.88 1.04 1.49 1.73 2.07 3.17 3.19 5.66 6.52 9.61 10.34 11.40 12.25 17.80 14.36 12.81 11.07 9.58 7.24 4.47 3.77 2.80

38.13 88.27 140.54 208.83 320.96 440.09 580.66 775.14 880.85 980.41 1049.29 113.59 1173.16 1230.65 1271.89 1348.31 1403.09 1434.68 1463.22 1489.73 1513.12 1535.02 1554.37 1566.77

19.64 50.14 52.27 68.29 112.13 119.13 140.57 194.48 105.71 99.56 68.88 64.30 59.57 57.49 41.24 76.42 54.78 31.59 28.54 26.51 23.39 21.90 19.35 12.40

22.61 30.52 40.18 50.32 63.75 103.28 147.84 194.69 227.15 258.01 286.43 312.94 335.67 356.87 376.11 394.29 411.45 416.78 430.99 443.44 454.31 463.34 470.27 475.63

4.22 7.91 9.66 10.14 13.43 39.53 44.56 46.85 32.46 30.86 28.42 26.51 22.73 21.20 19.24 18.18 17.16 5.33 24.21 12.45 10.87 9.03 6.93 5.36

Av. wt. = average weight; Inc. = increase The experiment was started with the initial average fork lengths of 51.68 ± 0.06 and 81.32 ± 0.07 mm of C. mrigala and C. carpio, respectively (Table 3). At the end of experiment C. mrigala and C. carpio attained the average fork lengths of 297.57 ± 3.21 and 208.32 ± 2.34 mm; 426.47 ± 4.51 and 262.48 ± 3.52 in mm treated and control ponds, respectively (Table 3). The maximum increase in fork length in C. mrigala was recorded during the 11th fortnight as 25.45 closely followed by 20.69 mm during 10th fortnights, respectively. As regards C. carpio it showed a maximum increase in fork lengths in the ponds supplemented with test diet 48.17 mm during 2nd followed by 39.52 mm during 1st, respectively. The growth rate, in the beginning of the experiment in this study was probably due to the fact both fish species are bottom feeders. In this study, best growth of carps was achieved within a temperature range of 20.53-30.41 °C. The present results do not support the findings of Barthelmes and Jahnichen (1978), who reported list growth of silver carp

(Hypophthalmichthvs molitrix) at water temperatures ranging from 18.00 to 22.00 °C. It is evident from the above results that growth is dependent upon food consumption and surplus energy after meeting the energy requirements of basal metabolism and activity of the fish. Feeding activity and food requirements and consumption patterns are greatly influenced by temperature. It appears net efficiency was more at these temperatures. Varghese et al. (1980) reported that silver carp (Hypophthalmicthys molitrix), grass carp (Ctenopharyngodon Idella) male common carp (Cyprinus Carpio), rohu (Labeo rohita) and mrigal (Cirrhina mrigala), responded to artificial feed comprising of rice bran: oil cake 1:1 at 2% body weight, were in accordance with the results of the present study. It was interesting to note that although both C. mrigala and C. carpio is bottom feeder but performed better with test diet which contradicted the results of Chakarabarty et al. (1976).

Table 3: Fortnightly average increase in fork length (mm) of Cirrhinus mrigala and Cyprinus carpio in control and feed supplemented pond

Fortnight 1 2 3 4 5 6 7 8 9 10 11 12 13 14

Cirrhinus mrigala Feed Supplemented Control pond pond AV. F. L Inc. AV. F.L Inc. (mm) (mm) (mm) (mm) 57.02 5.34 54.83 3.21 60.15 3.13 59.16 4.33 63.27 3.12 63.29 4.13 70.39 7.12 68.35 5.06 79.46 9.07 74.92 6.57 90.54 11.06 81.72 7.20 109.78 19.24 89.12 7.40 125.22 15.44 98.86 9.74 142.99 17.77 108.05 9.19 163.68 20.69 113.87 5.82 188.13 25.45 119.23 5.36 201.52 13.39 132.38 13.15 216.98 15.46 145.49 13.11 232.27 15.29 158.05 12.56

Cyprinus carpio Feed supplemented Control pond AV. F.L Inc. AV. F.L Inc. (mm) (mm) (mm) (mm) 120.84 39.52 87.88 6.56 169.01 48.17 101.42 13.54 198.19 29.18 123.53 22.11 215.38 17.19 125.15 1.62 241.95 26.57 136.73 11.58 275.29 33.34 162.02 25.29 310.68 35.39 189.21 27.19 332.44 21.76 204.65 15.44 340.17 7.73 209.99 5.34 359.75 19.58 225.46 15.47 364.86 5.11 234.81 9.35 368.09 3.23 239.56 4.75 373.33 5.24 243.12 3.56 384.99 11.66 246.74 3.63

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WWW.AFINIDAD.ORG 15 16 17 18 19 20 21 22 23 24

247.44 250.11 264.33 273.72 279.85 285.06 289.23 294.58 296.25 297.57

15.17 2.67 14.22 8.39 6.13 5.21 4.17 5.35 1.67 1.32

160.74 171.52 179.82 186.11 192.67 197.31 201.41 204.96 206.55 208.32

2.69 10.78 8.30 6.29 6.56 4.64 4.10 3.55 1.59 1.77

388.62 400.21 408.49 412.32 415.54 417.83 420.13 423.68 425.41 426.47

3.63 11.59 8.28 3.83 3.12 2.39 2.30 3.35 1.73 1.06

248.23 250.62 252.92 253.42 256.71 256.71 258.37 259.86 261.17 262.48

1.48 2.39 2.30 1.50 1.12 1.12 1.66 1.49 1.31 1.21

A.V. F.L= Average fork length; Inc.= increase Length-weight Relationship and Condition Factor Length-weight relationship is of great importance in fisheries. This relationship of fish has often been studied biologically. Length is considered as an independent variable while weight as a dependent one. The main objective of the length - weight relationship is to determine the variation from the expected weight of fish or a group of a fish to express the condition of fish in numerical terms (degree of well-being, relative robustness, plumpness or fatness). This relationship is called the coefficient of condition or condition factor. It was calculated from length-weight data for comparison of the condition of fish under the influence of various treatments. According to LeCren (1951) the length-weight relationship would first be calculated as the logarithmic formula: log W = log c + n log L The length-weight equations were fitted to the mean values obtained from the weights (g) and fork length of sample fishes of each species at each fortnight. The values of c and n were obtained

through the computer. From these values calculated weights were determined for a known mean fork lengths for which the mean observed weight were also known. In the present study the relative condition coefficient (Kn) which is independent of the units of measurements has been obtained by the formula: Kn = W / w Where “W” is observed weight and “w” is the calculated weight of fish. The regression equations for fork length-weight relationships of fish species is presented in Table 4. The high values of “r” for regression equations at each treatment level indicated reasonable precision of these equations for each fish species. Lengthweight indicates that both the fish species did not deviate from the general trend of such relationship feed supplementation. However, the differences were remained statistically significant at fortnight levels, indicating that fish deviated from the general trend of its length-weight relationship with age and season.

Table 4: Fork Length –Weight relationship for C. mrigala and C. carpio in feed supplemented and control pond Fish Species

Treatment

Regression Equation

r

C. mrigala Feed Supplementation log W= -4.62+2.929 log F.L 0.996 (0.056) Control log W= -5.44+3.302 log F.L 0.999 (0.028) C. carpio Feed Supplementation log W= -4.70+3.003 log F. L 0.999 (0.033) Control log W= -4.37+2.899 log F. L 0.996 (0.059) Values within brackets are standard errors; W: fish weight; F.L: fork length

After one year experimental period all the ponds were harvested for final fish yield. Total production for C. mrigala with feed supplementation and without any additives (control) ponds were calculated 8085.70 and 28786.81 g, respectively. Fish meal is one of the most expensive ingredients in prepared fish diets. Fish nutritionists have tried to use less expensive plant protein sources to partially or totally replace fish meal. The effects of plant protein meals and concentrates on the growth of rainbow trout have been reported by various workers (Gatlin et al., 2007; Lee et al., 2006; Lim et al., 2008; Mahboob and Sheri, 1997). Finding alternative protein sources to replace fish meal in fish feed is important if the growth of the

aquaculture industry is to be sustained (Francis et al., 2001; Tacon, 2003). It has been observed in this study that both the fishes performed significantly better in terms of increase in the weight and length compared to control. The better conversion efficiency feed into fish flesh in C. mrigala and C. carpio is due to their feed habit. Both the fish species are surface feeder (Mahboob and Sheri, 2002) CONCLUSION The results from the current study growth trial exhibit that C. mrigala and C. carpio have the potential to plant based diet efficiently and substantially enhanced the growth of these fish

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WWW.AFINIDAD.ORG species. C. carpio performed better as compared to C. mrigala. The findings of the present study suggest that fish meal can be replaced easily with alternative plant protein sources like cottonseed meal, maize gluten meal and sesamean oil cake. ACKNOWLEDGEMENTS This project was supported by Deanship of Scientific Research, College of Science, Research Center, King Saud University, Saudi Arabia. REFRENCES Alam AK, Maughan E, Matter WJ. Growth response of indigenous and exotic carp species to different protein sources in pelleted feeds. Aqua Res 1996;27:673-679. Barrows FT, Gaylord TG, Sealey WM, Porter L, Smith CE. Supplementation of plant-based diets for rainbow trout, Oncorhynchus mykiss with macrominerals and inositol. Aquaculture Nutrition 2010;16:654–661. Barthelmes D, Jahnichen H. Diet selection and feed intake of silver carp of 3 or 4 summers. Zeitschrift fur die Binnenfischerei der DDR 1978;25:331-334. Bhosale SV, Bhilave MP, Nadaf SB. Formulation of Fish Feed using Ingredients from Plant Sources. Res J Agri Sci 2010;1:284-287. Chakrabarty PC, Basak PK, Sarkar G, Ghosh G. Use of a new artificial feed in carp seed raising. In: Symposium on Inland Aquaculture 1979;pp:110. Cheng ZJ, Hardy RW, Usry JL. Effects of lysine supplementation in plant protein-based diets on the performance of rainbow trout (Oncorhynchus mykiss) and apparent digestibility coefficients of nutrients. Aquaculture 2003;215:255–265. FAO. The State of World Fisheries and Aquaculture. Rome 2010;pp:218. Francis G, Makkar HPS, Becker K. Antinutritional factors present in plant-derived alternate fish feed ingredients and their effects in fish. Aquaculture 2001;199:197-227. Gatlin DM, Barrows FT, Brown P, Dabrowski K, Gaylord TG, Hardy RW, Herman E, HU G, Krogdahl A, Nelson R, Overturf K, Rust M, Sealey W, Skonberg D, Souza EJ, Stone D, Wilson R, Wurztel E. Expanding the utilization of sustainable plant products in aquafeeds: a review. Aquaculture Research 2007;38:551–579. Glencross BD, Carter CG, Duijster N, Evans DR, Dods K, Mccafferty P, Hawkins WE, Maas R, Sipsas S. A comparison of the digestibility of a range of lupin and soybean protein products when fed to either Atlantic salmon (Salmo salar) or rainbow trout (Oncorhynchus mykiss). Aquaculture 2004;237:333–346. Hora SL, Pillay TVR. Handbook on Fish Culture in the Indo-Pacific region. FAO Fish Biol 1962;(14):204. Huisman EA, Klei Breteler JGP, Vismans MM, Kanis E. Retention of energy, protein, fat and ash in growing carp (Cyprinus carpio) under different

feeding and temperature regimens. In: Halver IE, Tiews K (Eds.), Finfish Nutrition and Fish Feed Technology. Vol. I. Heeneman Veriagsgesells Chaft, Berlin 1979;1:175-188. Javed M. Growth performance and meat quality of major carps as influenced by pond fertilization and feed supplementation. Ph.D. Thesis, Agri. Univ., Faisalabad 1988. Lee KJ, Rinchard J, Dabrowski K, Babiak I, Ottobre JS, Christensen JF. Long term effects of dietary cottonseed meal on growth and reproduction performance of rainbow trout: three-year study. Anim Feed Sci Tech 2006;126:93–106. Lim C, Webster CD, Lee CS. Alternative protein sources in aquaculture diets. The Haworth Press, New York, NY 2008;pp:571-281. Mahboob S, Sheri AN. Growth performance of major, common and some Chinese carps as influenced by pond fertilization and feed supplementation in composite culture system. J Aqua Trop 1997;12:201-207. Mahboob S, Sheri AN. Influence of fertilizers and artificial feed on the total dressing losses, meat: bone ratios, specific gravity and density of fish and regression studies of dry weight of planktonic biomass on physico-chemical parameters. J Aqu Trop 2002;17:43-58. Naylor R, Hardy R, Bureau D, Chiu A, Elliott M, Farrell A, Forster I, Gatlin D, Goldberg R, Hua K, Nichols P. Feeding aquaculture in an era of finite resources. Proc Nat Acad Sci 2009;106l: 15103– 15110 Varghese TJ, Singit GS, Keshavanath P, Reddy PK, Vasudevappa C. Composite fish culture a case study. Mysore J Agri Sci 1980;14(2):232-236.

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