Necessity of Mineral Supplement to Fish Meal Based Red Sea Bream Feed*1

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1 SUISANZOSHOKU 46(4), (1998) Necessity of Mineral Supplement to Fish Meal Based Red Sea Bream Feed*1 Shuichi SATOH*2, Ryotaro ISHIDA*2, Toshio TAKEUCHI*2, Takeshi WATANABE*2, and Tadahisa SEIKAI*3 (Accepted September 9, 1998) Abstract: A feeding experiment was conducted to investigate whether it was necessary to supplement minerals to fish meal based feed for red sea bream or not. Phosphorus (P), magnesium (Mg), zinc (Zn), or manganese (Mn) was excluded respectively from the complete mineral mixture. The mineral mixtures were formulated to the fish meal based experimental diet. The experimental diets were fed to the red sea bream (2.4 g on average) for nine weeks. The growth was lowest in the fish fed the diet without P supplementation, the growth of the fish fed the diet without Zn supplement was significantly lower than the control group. However, influence of excluding Mg or Mn supplementation on the growth was very small. Zn content in vertebrae was lowest in the fish on the diet without Zn supplement, and Mn content in vertebrae was also lowest in the fish on the diet without Mn supplement. When red sea bream were fed the diet without P, Mg, Mn, or Zn, their growth and mineral composition in carcass were influenced. The results of the experiment suggested that availability of minerals contained in fish meal for red sea bream was not high, and it was necessary to supply minerals even to fish meal based feed. Key words: Diet; Red sea bream; Fish meal; Mineral supplement Recent investigations on mineral nutrition in fresh water fish such as rainbow trout, common carp, and tilapia have demonstrated that the availability of minerals contained in fish meal varies amongst fish species1-3). The optimum supplemental levels vary according to the kind of minerals and the digestive systems of fish species. It was reported that juvenile red sea bream ( g), a major aquaculture species in Japan, requires only phosphorus (P) and iron (Fe) at the levels 6.8 mg/g and 150 ƒêg/g, respectively4,5), even in semi-purified casein based diet and that trace element supplementation to the casein diet is not essential when aluminum (Al), zinc (Zn), manganese (Mn), copper (Cu), cobalt (Co), and iodine (I) exist at levels exceeding 2 mg/kg, 24 mg/kg, 18 mg/kg, 5 mg/kg, 4 mg/kg, and 0.1 mg/kg, respectively6). Excluding the supplementation of some minerals which are normally added to fish meal based feed of red sea bream did not produce any negative effects in the case of fish weighing 45g*4. Therefore, necessary it was considered that it may not be to supply minerals to red sea bream feed which contained fish meal as the main component. Satoh et al. 7) reported that mineral contents of whole body of rainbow trout becomes hardly change after body weight reached 2 g, and recommended to conduct the feeding experiment on smaller than 2 g. Thus, this study was con- *1 Mineral Nutrition in Fish-XXXIṾ *2 Department of Aquatic Biosciences, Tokyo University of Fisheries, Konan, Minato, Tokyo , Japan. *3 Fisheries Experimental Station, Kyoto University, Maizuru, Kyoto 625, Japan. *4 M. Furuichi, S. Kosho, S. Matsui, and C. Kitajima: Necessity of mineral supplement to marine fish feed. Abstract of Annual Meeting of Japanese Society of Fisheries Science, p. 533, 1994.

2 536 S. Satoh, R. Ishida, T. Takeuchi, T. Watanabe, and T. Seikai ducted to investigate whether mineral supplement was necessary to fish meal based feed for very small red sea bream. The juveniles of red sea bream were fed the fish meal based diet supplemented with or without minerals. Experimental Diets Materials and Methods The formulations of the experimental diets are shown in Table 1. White fish meal was used as a sole protein source. Diet 1 was a control supplemented with a complete mineral mixture which fulfils mineral requirements of fresh water fish, such as rainbow trout and carp at a level of 5%. Diets 2 to 5 were supplemented with the mineral mixtures in which Zn, Mn, magnesium (Mg), or P was excluded, respectively. The other contents of the diets 2 to 5 were the same as that of the diet 1(control diet). The mineral compositions of the experimental diets are shown in Table 2. Crude protein and lipid contents were the same among the experimental Table 1. Composition of the experimental diets used for red sea bream (%) diets, which were 45% and 20%, respectively. Crude ash content was lowest in the diet (diet 5) without phosphates supplementation. Calcium (Ca), P, potassium (K), and sodium (Na) contents were also lowest in diet 5, but the contents were similar in the other diets. Mg, Zn, and Mn contents were lower in the diet without Mg, Zn or Mn supplementation, respectively, and in the supplemented groups the contents of these elements were similar to each other. The quantum of P derived from fish meal in diet 5 seemed to satisfy the P requirements of red sea bream5). Feeding Experiment Juvenile red sea bream Pagrus major were used as the experimental animals. Red sea bream larvae were transported from the Energy Research Institute of Kansai Denryoku K. K. to the Fisheries Experimental Station of Kyoto University and kept on a commercial red sea bream feed until the feeding experiment. The juvenile red sea bream (2.4 g on average) were divided into five tanks of 25 fish each, and the diets shown in Table 2 were fed to satiety for nine weeks. Water temperature ranged 20 to 24 Ž. Water was supplied at the rate of 0.8 to 1.0 l/m. At the termination of feeding experiment, all the fish were weighed individually, and kept in a freezer until analyses of proximate and mineral compositions in whole body and vertebrae. Preparation of samples and analytical conditions for proximate and mineral compositions were the same as those described *Same as the pre vious paper7). previously7). Final body weight of the fish in each tank was subjected to one-way analysis of variance and the least significant difference test to determine difference in means (p<0.05)8). Table 2. Mineral composition of the experimental diets used for red sea bream

3 Necessity of Mineral Supplement to Red Sea Bream Feed 537 Table 3. Effect of excluding certain mineral supplementation from fish meal based diet on the growth performance of red sea bream * 1 g gro wth/g feed. * 2 Values within a column followed by the same superscript letter are not significantly different (p>0.05). Table 4. Effect of excluding certain mineral supplementation from fish meal based diet on the proximate and mineral compositions of whole body of red sea bream Feed efficiency showed a trend similar to Fig. 1. Influence of excluding certain mineral supplementation from fish meal based diet on the growth of red sea bream. Results The results of feeding experiment are shown in Table 3 and Fig. 1. Growth became lower from the third week in the fish fed the diet without P or Zn supplement compared to the rest, and at termination it was lowest in the fish on the P unsupplemented diet, followed by the Zn unsupplemented diet, being significantly lower than the control group. However, the growth of the group fed the diet without Mn supplement was equivalent to the control group. growth. Survival was not influenced by dietary treatments during the feeding period. External symptoms of mineral deficiency observed in rainbow trout and common carp such as eye lens cataracts and short body dwarfism were not recognized in the current study. Proximate and mineral compositions of whole body are shown in Table 4. Moisture content was slightly higher in the fish fed the diet without P or Mn supplementation than that of the control group. Lipid content was also lower in the fish fed the diet without Mg or Mn supplementation than that of the control group. Ash content was markedly lower in the fish on the diet without P supplementation than the other groups. There were no marked differences in Ca, Na, K, Mg, and Fe contents of whole body amongst any dietary groups. Zn and Mn contents were

4 538 S. Satoh, R. Ishida, T. Takeuchi, T. watanabe, and T. Seikai lower in the fish fed the diets without Zn or Mn supplement respectively than the fish on the control diet. P content was higher in the fish on the diet without Mn or Mg supplements than that of the control group. Mineral compositions of vertebrae are shown in Table 5. There were no marked differences in P, Ca, and Na contents amongst the treatments. Zn content was lowest in the vertebrae fish fed the diet without of the Zn supplementation amongst the experimental groups, and was also lower in the fish on the diets without P or Mg than the control group. Mn content was much lower in the fish fed diet without Mn supplementation than the control group, and Mn content in the fish fed the diet without P or the Mg supplementation was slightly lower than the control. The Mn content of the fish on the control diet was much higher than that was reported for rainbow trout9). Discussion It was reported that supplementation with mg P/g diet was required by red sea bream when the fish were fed the casein based diets supplemented with graded levels of P from 0 to 9.1 mg/g by mono-basic sodium phosphate5). The fish meal based diet without P supplementation used in the current study contained 20 mg P/g derived from the fish meal satisfying the P requirement. However, these fish had low growth and feed efficiency. The supplementation with 8 mg P improved the growth and increased ash content. Ogino et al.10) reported that rainbow Table 5. Effect of excluding certain mineral supplementation from fish meal based diet on the mineral compositions of vertebrae of red sea bream trout was able to absorb 60-70% of P contained in fish meal, whereas common carp could absorb only 0-10% of P from fish meal. Yone and Toshima11) also reported that black sea bream, Acanthopagrus shlegeli, could utilize 30% of P contained in fish meal. Red sea bream is a species closely related to black sea bream, and seems to possess an absorption pattern similar to black sea bream. Recently, Shimeno and coworker12,13) reported that the requirement of yellowtail, Seriola quinqueradiata, for dietary P was 6.7 mg/g diet, which could attain by supplementing P at a level of 3.4 mg/g to fish meal based feed. They also suggested that yellowtail could utilize P contained in fish meal partially. As the red sea bream fed the fish meal based diet without P supplementation showed lower growth than that on the diet with P supplementation the current study, it seemed in that red sea bream could not absorb P in fish meal completely. Thus, it is necessary to supply P even to fish meal based diet. It was reported that tricalcium phosphate derived from fish meal inhibits the absorption of Zn and Mn in fresh water fish such as rainbow trout2,3). In the current study, the external Zn deficiency symptom such as eye lens cataracts and short body dwarfism was not induced by the diet without supplemental Zn. However, the fish fed the diet without Zn supplement lower growth, induced feed efficiency, and Zn contents in carcass. Thus, it is suggested that the availability of Zn in fish meal to red sea bream is not high, and it is necessary fish meal based diet for red sea bream. to supplement Zn to The diet without Mn supplement did not lower the growth of red sea bream, however it induced lower Mn contents in carcass. When the fish meal based diet without Mn supplement was fed to common carp, it lowered growth, decreased Mn and Zn contents, and induced short body dwarfism and eye lens cataractsl4). In current study, for sea bream fed the diet without Mn supplement, decreased, Mn content in carcass but Zn content was not influenced. Further, Mn content of red sea bream was higher than that of the rainbow trout and common carp. The external deficiency symptoms

5 Necessity of Mineral Supplement to Red Sea Bream Feed 539 were not observed in this study, however, it is recommended to supply Mn to red sea bream feed as well. As for Mg, significant influence was not observed in the growth of the fish fed the diet without Mg supplement. Sakamoto and Yone15) also reported that it was not necessary to supplement Mg to red sea bream diet when they were fed the semi-purified diet. However, Zn content was lowered in vertebrae of the fish fed the diet without Mg supplementation, similar to the results of rainbow trout9). Satoh et al.16) reported that rainbow trout required Mg supplement even for fish meal based feed at a level of 0.5 mg/g diet. Therefore, it is suggested that Mg supplement is necessary for red sea bream feed too. The fish meal based diet contains apparently sufficient amount of minerals derived from the ingredients, and those levels of minerals seemed to satisfy the mineral requirement of red sea bream. However, when red sea breams were fed the diet without P, Mg, Mn, or Zn, the growth and mineral content in whole body and vertebrae were influenced. Filk et al.17) reported that Ca is actively absorbed through gill epithelium in freshwater, whereas Ca is transported across the intestinal epithelium in seawater. There is no information available concerning the other minerals in seawater. If the minerals are transported as same as Ca, seawater fish may not absorb the minerals from ambient water. The results of the experiment demonstrated that the availability of minerals contained in fish meal to red sea bream was not sufficient enough, necessitating supplementation to improve performance of this fish, which is one of the major species cultivated in sea water. The optimum supplemental levels of minerals to fish meal based diet of red sea bream will be described soon. Acknowledgments We express here our sincere thanks to the Energy Research Institute of Kansai Denryoku K.K., which kindly provided juvenile red sea bream. Our thanks also extend to Dr. Viswanath Kiron, Tokyo University of Fisheries, who kindly read the manuscript and gave valuable suggestions. References 1) Watanabe, T.,S. Satoh, and T.Takeuchi (1988): Availability of minerals in fish meal to fish. Asian Fisheries Science, 1(2), ) Satoh, S. (1994): Studies on the availability of trace elements to fish. Nippon Suisan Gakkaishi, 60(2), ) Watanabe, T., V. Kiron, and S. Satoh (1997): Trace minerals in fish nutrition. Aquaculture, 151(1-4), ) Sakamoto, S. and Y. Yone (1978): Requirement of red sea bream for dietary iron-ii. Bull. Japan. Soc. Sci. Fish., 44(3), ) Sakamoto, S. and Y. Yone (1978): Effect of dietary phosphorus level on chemical composition of red sea bream. Bull. Japan. Soc. Sci. Fish., 44(3), ) Sakamoto, S. and Y. Yone (1979): Requirement of red sea bream for dietary trace elements. Bull. Japan. Soc. Sci. Fish., 44(12), ) Satoh, S., T. Takeuchi, and T. Watanabe (1987): Changes of mineral compositions in whole body of rainbow trout during growing stages. Nippon Suisan Gakkaishi, 53(2), ) Duncan, D. B. (1955): Multiple-range and multiple F tests. Biometrics, 11, ) Satoh, S., H. Yamamoto, T. Takeuchi, and T. Watanabe (1983): Effects on growth and mineral composition of rainbow trout of deletion of trace elements or magnesium from fish meal diet. Bull. Japan. Soc. Sci. Fish., 49(3), ) Ogino, C., L. Takeuchi, H. Takeda, and T. Watanabe (1979): Availability of dietary phosphorus in carp and rainbow trout. Bull. Japan. Soc. Sci. Fish., 45(12), ) Yone, Y. and N. Toshima (1979): The utilization of phosphorus in fish meal by carp and black sea bream. Bull. Japan. Soc. Sci. Fish., 45(6), ) Shimeno, S. (1991): Yellowtail, Seriola quinqueradiata, in ghandbook of Nutrient Requirements of Finfish h(ed. by R. P. Wilson), CRC Press, Boston, pp ) Shimeno, S., A. Hashimoto, and M. Ukawa (1994): Effects of supplemental levels of dietary monopotasium phosphate on growth, feed conversion, and body composition of fingerling yellowtail. Suisanzoshoku, 42(2), ) Satoh, S., H. Yamamoto, T. Takeuchi, and T. Watanabe (1983): Effects on growth and mineral composition of carp of deletion of trace elements or magnesium from fish meal diet. Bull. Japan. Soc. Sci. Fish., 49(3), ) Sakamoto, S. and Y. Yone (1979): Requirement of red sea bream for Mg. Bull. Japan. Soc. Sci. Fish., 45(1), ) Satoh, S., T. Takeuchi, and T. Watanabe (1991):

6 540 S.Satoh, R.Ishida, T.Takeuchi, T.Watanabe, and T.Seikai Availability of manganese and magnesium contained in white fish meal to rainbow trout Oncorhynchus mykiss. Nippon Suisan Gakkaishi, 57(1), ) Flik, G., P. M. Verbost, and S. E. Wendelaar Bonga (1995): Calcium transport processes in fishes, in gcellular and Molecular Approaches to Fish Ionic Reg-ulation. Fish Physiology, Vol.14 h(ed. by C. M. Wood T. and J. ShuA

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