Effect of Partial Replacement of Fish Meal by Bacillus sp-Fermented Soybean Meal on Growth Performance, Immunity, Hepatopancreas Microbiota and Disease Resistance in Pacific White Shrimp (Litopenaeus vannamei)

Main Article Content

Kanokwan Cherdkeattipol
Niti Chuchird
Duenrut Chonudomkul
Wichien Yongmanitchai
Phongchate Pichitkul


This study examined the effect of substituting a diet containing 20% fish meal (FM) and 10% soy bean meal (SBM) with Bacillus-fermented soybean meal (FSBM) on growth, immune response, microbial community in the hepatopancreas, and disease prevention in Pacific white shrimp. Shrimp were fed diets with four levels of FSBM (0%, 15%, 20% and 25%) to replace FM and SBM. After 60 days of the feeding trial, no difference was found among treatments for specific growth rate (1.62-1.66 g), survival rate (88.08-93.64 %), feed conversion ratio (FCR) (1.31-1.41), or protein efficiency ratio (88.08-93.46). However, shrimp fed 25% FSBM showed a significant improvement in total hemocyte count (THC), phagocytic activity, and phenoloxidase activity (PO) compared with other groups. In the hepatopancreatic microbiota, we identified six phyla, of which Bacteroidetes was the most dominant phylum in the three groups fed with FSBM. In contrast, Firmicutes was dominant in the control group. After a challenge test with Vibrio parahaemolyticus, shrimp fed 25% FSBM had a significantly higher average survival rate compared with other experimental groups infected with V. parahaemolyticus. The histopathology of the hepatopancreas of shrimp from this group showed fewer signs of bacterial infection than other groups infected with V. parahaemolyticus. This study indicates that FSBM at the concentration of 25% can enhance immune response and tolerance to pathogenic V. parahaemolyticus in the Pacific white shrimp.


Download data is not yet available.

Article Details



1. Amaya, E.A., D.A. Davis and D.B Rouse. 2007. Replacement of fish meal in practical diets for the Pacific white shrimp (Litopenaeus vannamei) reared under pond conditions. Aquaculture 272: 466‒476.

2. Bell, T.A. and D.V. Lightner. 1998. A Handbook of Normal Penaeid Shrimp Histology. World Aquaculture Society, Baton Rouge, Los Angeles. 114 pp.

3. Cheng, W., C.S. Chiu, Y.K. Guu, S.T. Tsai and C.H. Liu. 2013. Expression of recombinant phytase of Bacillus subtilis E20 in Escherichia coli HMS174 and improving the growth performance of white shrimp, Litopenaeus vannamei, juveniles by using phytase-pretreated soybean meal-containing diet. Aquaculture Nutrition 19: 117‒127.

4. Chen, J.N., X.Q. Li, H.B. Xu, W.T. Sun and X.J. Leng. 2017. Substitute of soy protein concentrate for fish meal in diets of white shrimp (Litopenaeus vannamei Boone). Aquaculture International 25(3): 1303‒1315.

5. Decamp, O., D.W. Moriarty and P. Lavens. 2008. Probiotics for shrimp larviculture: review of field data from Asia and Latin America. Aquaculture Research 39: 334‒338.

6. Ding, Z., Y. Zhang, J. Ye, Z. Du and Y. Kong. 2015. An evaluation of replacing fish meal with fermented soybean meal in the diet of Macrobrachium nipponense: Growth, nonspecific immunity, and resistance to Aeromonas hydrophila. Fish and Shellfish Immunology 44: 295‒301.

7. Edgar, R.C., B.J. Haas, J.C. Clemente, C. Quince and R. Knight. 2011. UCHIME improves sensitivity and speed of chimera detection. Bioinformatics 27(16): 2194‒2200.

8. Fan, P., P. Liu, P. Song, X. Chen and X. Ma. 2017. Moderate dietary protein restriction alters the composition of gut microbiota and improves ileal barrier function in adult pig model. Scientific Reports 7: 43412. DOI: 10.1038/srep43412.

9. Farzanfar, A. 2006. The use of probiotics in shrimp aquaculture. FEMS Immunology and Medical Microbiology 48: 149‒158.

10. Hamidoghli, A., M.S. Djaballah, S. Maamri, A. Hamdi, I. Souffi N.W. Farris and S.C. Bai. 2020. Effect of different dietary plant protein sources as fishmeal replacers in juvenile white leg shrimp, Litopenaeus vannamei. Fisheries and Aquatic Sciences 23: 2‒6.

11. Hong, K.J., C.H. Lee and S.W. Kim. 2004. Asperagillus oryzae GB-107 fermentation improves nutritional quality of food soybeans and feed soybean meals. Journal of Medicinal Food 7: 430‒435.

12. Itami, T., Y. Takahashi, E. Tsuchihira, H. Igasu and M. Kondo. 1994. Enhancement of discase resistance of Kuruma Prawn Penaeus japonicas and increase in phagocytic activity of prawn hemocytes after oral administration of β-1,3-glucan (Schizophyllan). Proceedings of the 3th Asian Fisheries Forum Singapore 1992: 375‒378.

13. Kiers, J.L., A.E.A. Van laeken, F.M. Rombouts and M.J.R. Nout. 2000. In vitro digestibility of Bacillus fermented soya bean. International Journal of Food Microbiology 60: 163‒169.

14. Kim, M., E. Min, J. Kim, J. Koo and J. Kang. 2015. Growth performance and immunological and antioxidant status of Chinese shrimp, Fennerpenaeus chinensis reared in bio-floc culture system using probiotic. Fish and Shellfish Immunology 47: 141‒146.

15. Krummenauera, K., L. Poerscha, L.A. Romanob, R. Gabriele, P. Encarnação and W. Wasielesky. 2014. The effect of probiotics in a Litopenaeus vannamei biofloc culture system infected with Vibrio parahaemolyticus. Journal of Applied Aquaculture 26: 370‒379.

16. Lin, Y.H. and J.J. Mui. 2017. Comparison of dietary inclusion of commercial and fermented soybean meal on oxidative status and non-specific immune responses in white shrimp, Litopenaeus vannamei. Fish and Shellfish Immunology 63: 208‒212.

17. Liu, C.H., Y.L. Shiu and J.L. Hsu. 2012. Purification and characterization of trypsin from the pyloric ceca of orange-spotted grouper, Epinephelus coioides. Fish Physiology and Biochemistry 38(3): 837‒848.

18. Miao, S., J. Zhu, C. Zhao, L. Sun, X. Zhang and G. Chen. 2017. Effect of C/N ratio control combined with probiotics on the immune response, disease resistance, intestinal microbiota and morphology of giant freshwater prawn (Macrobrachium rosenbergii). Aquaculture 476: 125‒133.

19. Nayak, S., C. Limsuwan, N. Chuchird and S. Pungpang. 2012. A study on the effect of Bacillus spp. to control the pathogenic bacteria in aquaculture. Kasetsart University Fisheries Research Bulletin 36(2): 1‒12.

20. Nguyen, N.V., L. Hoang, T.V. Khanh, P.D. Hai and L.T Hung. 2017. Utilization of fermented soybean meal for fishmeal substitution in diets of Pacific white shrimp (Litopenaeus vannamei). Aquaculture Nutrition 24: 1092‒1100.

21. Nonwachai, T., W. Purivirojkul, C. Limsuwan, N. Chuchird, M. Velasco and A.K. Dhar. 2010. Growth, nonspecific immune characteristics, and survival upon challenge with Vibrio harveyi in Pacific white shrimp (Litopenaeus vannamei) raised on diets containing algal meal. Fish and Shellfish Immunology 29(2): 298‒304.

22. Rengpipat, S., S. Rukpratanporn, S. Piyatiratitivorakul and P. Mennasaveta. 2000. Immunity enhancement in Black tiger shrimp (Penaeus monodon) by a probiont bacterium (Bacillus S11). Aquaculture 191: 271‒288.

23. Rungrassamee, W., A. Klanchui, S. Maibunkaew, S. Chaiyapechara and P. Jiravanichpaisal. 2014. Characterization of intestinal bacteria in wild and domesticated adult black tiger shrimp (Penaeus monodon). PLoS ONE 9(3): e91853. DOI: 10.1371/journal.pone.0091853.

24. Rognes, T., T. Flouri, B. Nichols, C. Quince and F. Mahé. 2016. A versatile open source tool for metagenomics. PeerJ Computer Science 4: e2584. DOI: 10.7717/peerj.2584.

25. Sanjukta, S., A.K. Rai, A. Muhammed, K. Jeyaram and N.C. Talukdar. 2015. Enhancement of antioxidant properties of two soybean varieties of Sikkim Himalayan region by proteolytic Bacillus subtilis fermentation. Journal of Functional Food 14: 650‒658.

26. Shao, J., Z. Wei, H. Siyin and C. Yang. 2018. Partial replacement of fish meal by fermented soybean meal in diets for juvenile white shrimp (Litopenaeus vannamei). Aquaculture Nutrition 25: 145‒153.

27. Song, Y.S., J. Frias, C.M. Villaluenga, C.V. Valdeverde and E.G. de Mejia. 2008. Immunoreactivity reduction of soybean meal by fermentation, effect on amino acid composition and antigenicity of commercial soy products. Food Chemistry 108: 571‒581.

28. Sookying, D., D.A. Davis and F. Soller. 2013. A review of the development and application of soybean-based diets for Pacific white shrimp (Litopenaeus vannamei). Aquaculture Nutrition 19: 441‒448.

29. Tacon, A.G.J. and M. Metian. 2008. Global overview on the use of fish meal and fish oil in industrially compounded aquafeeds. Aquaculture 285: 146‒158.

30. Tanrattanapitak, N. and S. Pairohakul. 2018. Bacterial community in gut contents of the sea urchin Diadema setosum (Leske, 1778) and the ambient sediments from Sichang Island using metagenomics approaches. International Journal of Science 15(1): 117‒125.

31. Vaseeharan, B. and P. Ramasamy. 2003. Control of pathogenic Vibrio spp. by Bacillus subtilis BT23, a possible probiotic treatment for black tiger shrimp Penaeus monodon. Applied Microbiology 36(2): 83‒87.

32. Wangsoontorn, S., N. Chuchird, I. Wudtisin and A. Crook. 2018. The effect of substituting fish meal with fermented soybean meal on the growth performance and immune parameters of Pacific white shrimp (Litopenaeus vannamei). Journal of Fisheries and Environment 42(2): 32‒40.

33. Wu, S., G. Wang, E.R. Angert, W. Wang, W. Li and H. Zou. 2012. Composition, diversity and origin of the bacterial community in grass carp intestine. PLoS ONE 7(2): e30440. DOI: 10.1371/journal.pone.0030440.

34. Wu, S., T.H. Gao, Y.Z. Zheng, W.W. Wang and Y.Y. Cheng. 2010. Microbial diversity of intestinal contents and mucus in yellow catfish (Pelteobagrus fulvidraco). Aquaculture 303: 1‒7.

35. Yamamoto, T., Y. Iwashita, H. Matsunari, T. Sugita, H. Furuita, A. Akimoto, K. Okamatsu and N. Suzuki. 2010. Influence of fermentation conditions for soybean meal in a non-fish meal diet on the growth performance and physiological condition of rainbow trout Oncorhynchus mykii. Aquaculture 309: 173‒180.

36. Yatip, P., D.N.C. Teja, T.W. Flegel and C. Soowannayan. 2018. Extract from the fermented soybean product natto inhibits vibrio biofilm formation and reduces shrimp mortality from Vibrio harveyi infection. Fish and Shellfish Immunology 72: 348‒355.

37. Yao, W., C. Zhang, X. Li, M. He, J. Wang and X. Leng. 2019. The replacement of fish meal with fermented soya bean meal or soya bean meal in the diet of Pacific white shrimp (Litopenaeus vannamei). Aquaculture Research 51: 2400‒2409.

38. Zhanga, M., Y. SunaKe, C. Yua, Z. Zhoub, L. Chena and Z. Du. 2014. Characterization of the intestinal microbiota in Pacific white shrimp (Litopenaeus vannamei)fed diets with different lipid sources. Aquaculture 434: 449‒455.

39. Zokaeifar, H., J.L. Balcazar, C.R. Saad, M.S. Kamarudin, K. Sijam, A. Arshad and N. Nejat. 2012. Effect of Bacillus subtilis on the growth performance, digestive enzyme activity, immune gene expression and disease resistance of white shrimp (Litopenaeus vannamei) Fish and Shellfish Immunology 33: 683‒689.