Effect of different levels of Lactobacillus casei on growth performance and digestive enzymes activity of Shirbot (Barbus gryprus)


1 Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

2 Department of Biochemistry and Molecular Biology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

3 Khouzestan Fisheries Organization, Ahvaz, Iran


BACKGROUND: Lactic acid bacteria are the most common class of bacteria used in aquaculture as probiotic. ObjectiveS: In this study the effects of various levels of Lactobacillus casei on the growth performance and digestive enzymes activity of juvenile Shirbot were evaluated. Methods: Four hundred-eighty juvenile Shirbot weighing 40 g were divided randomly in four treatments (in triplicate). Treatments of A, B and C were fed with 5×106 CFU g-1, 5×107 CFU g-1, 5×108 CFU g-1, respectively for 60 days. Control group was fed with free-probiotic diet. After the period, treatments were fed with free-probiotic diet for 15 days. Growth indices and digestive enzymes were examined on days 0, 30, 60 and 75. Results: In the Treatment B, Specific Growth  Ratio, Daily Weight Growth and Relative Growth Rate, after 30 days from the beginning of experiment improved considerably which, compared to control group had significant difference (p<0.05). Activity of chymotrypsin enzyme in treatment of group B after 30 days and similarly, trypsine in treatment of group C after 30 and 60 days, were increased significantly compared to control group (p<0.05). In the other points of sampling and enzymes, considerable difference was not seen (p>0.05). Conclusions: Results showed that 5×107 CFUg-1 of Lactobacillus casei for 30 days and 5×108 CFUg-1 for 60 days, are the best doses of probiotic.


Abd El-Rhman, A.M., Khattab, Y.A., Shalaby, A.M. (2009) Micrococcus luteus and Pseudomonas spcies as probiotics for promoting the growth performance and health of Nile tilapia, Oreochromis niloticus. Fish Shellfish Immunol. 27: 175-180.
Al-Dohail, M.A., Hashim, R., Aliyu-Paiko, M. (2009) Effects of the probiotic, Lactobacillus acidophilus, on the growth performance, haematology parameters and immunoglobulin concentrationin African Catfish (Clarias gariepinus, Burchell 1822) fingerling. Aquac Res. 40: 1642-1652.
Aly, S.M., Ahmed, Y., Ghareeb, A., Mohamed, M.F. (2008) Studies on Bacillus subtilis and  Lactobacillus acidophilus, as potential probiotics, on the immune response and resistance of  Tilapia nilotica (Oreochromis niloticus) to challenge infections. Fish Shellfish Immunol. 25: 128-136.
Balcazar, J.L., Blas, I., Ruiz-Zarzuela, I., Cunningham, D., Vandrell, D., Muzquiz J.L. (2006) Review: The role of probiotics in aquaculture. Vet Microbiol. 114: 173-186.
Bernfeld, P. (1955) Amylases α and β. In: Methods in Enzymology. Colowick, P., Kaplan, N.O. (eds.). (1st ed.) Academic press, New York, USA. p. 149-157.
Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 72: 248-254.
Cahu, C.L., Zambonino Infante, J.L., Quazuguel, P., Le Gall, M.M. (1999) Protein hydrolysate vs. fish meal in compound diets for 10-day old sea bass Dicentrarchus labrax larvae. Aquaculture. 171: 109-11.
Chang, A.S.C., Hashim, R., Chow-Yang, L., Ali, A.B. (2002) Partial characterization and activities of proteases from the digestive tract of discus fish, Symphysodon aeguifasciata. Aquaculture. 203: 321-333.
Denev, S., Staykov, Y., Moutafchieva, R., Beev, G. (2009) Microbial ecology of the gastrointestinal tract of  fish and the potential application of probiotics and prebiotics in finfish aquaculture. Int Aquat  Res. 1: 1-29.
Erlanger, B.F., Kokowsky, N., Cohen, W. (1961) The preparation and properties of two new chromogenic substrates of trypsin. Arch Biochem Biophys  95: 271-278.
Giri, S.S., Sukumaran, V., Oviya, M. (2013) Potential probiotic lactobacillus plantarum VSG3 improves the growth, immunity, and disease resistance of tropical freshwater fish, labeo rohita. Fish Shellfish Immunol. 34: 660-666.
Gomez-Gil, B., Roque , A., Turnbull, J.F. (2000) The use and selection of  probiotic bacteria for use in the culture of  larvala quatic organisms. Aquaculture. 191: 259-270.
Hosseinifar, S.H., Mirvaghefi, A.R., Majazi Amiri, B., Khoshbavar Rostami, H.A., PourAmini, M., Darvish Bastami, K. (2010) The probiotic effects of Saccharomyces cerevisiae var. ellipsoideus on growth factors, survival, body composition and intestinal microbiota of Beluga juvenile (Huso huso). Iran Sci Fish J. 9: 55-66.
Hummel, B.C. (1959) A modified spectrophotometric determinations of chymotrypsin, trypsin and thrombin. Can J Biochem Physiol. 37: 1393-1399.
Kahkesh, F.B., Yooneszadeh Feshalami, M., Amiri, A., Nickpey, M. (2010) Effect of ovaprim, ovatid, HCG, LHRH-a2, LHRH-a2 + CPE and carp Pituitary in Benny (Barbus sharpeyi) artificial breeding. Glob Vet. 5: 209-214.
Kuz’mina, V., Shekovtsova, N., Bolobonina, V. (2010) Activity dynamics of proteinases and glycosidases of fish chyme with exposure in fresh and brackish water. Biol Bull. 37: 605-611.
Lara-Flores, M., Olivera-Novoa, M. ,Guzma´n-Me´ndez, B., Lo´pez-Madrid, W. (2003) Use of the bacteria Streptococcus faecium and Lactobacillus acidophilus, and the yeast Saccharomyces cerevisiae as growth promoters in Nile tilapia (Oreochromis niloticus). Aquaculture. 216: 193-201.
Lara-Flores, M., Olivera-Castillo, L., Olivera-Novoa, M.A. (2010) Effect of the inclusion of a bacterial mix (Streptococcus faecium and Lactobacillus acidophilus) and the yeast (Saccharomyces cerevisiae) on growth, feed utilization and intestinal enzymatic activity of Nile tilapia (Oreochromis niloticus). Int J Fish Aquac. 2: 93-101.
Lily, D.M., Stillwell, R.H. (1965) Probiotics: growth promoting factors produced by microorganisms. Science. 147: 747-748.
Makridis, P., Bergh, Ø., Skjermo, J., Vadstein, O. (2001) Addition of bacteria bioencapsulated in Artemia metanauplii to a rearing system for halibut larvae . Aquac Int. 9: 225- 235.
McCracken, V.J, Gaskins, H.R. (1999) Probiotics and the immune systems. In: Probiotics: A Critical Review. Tannock, G.W. (ed.). (1st ed.) Horizon Scientific Press, Norwich, England. p. 85-112.
Nikoskelainen, S., Ouwehand, A.C., Bylund, G., Salminen, S., Lilius, E.M. (2003) Immune enhancement in rainbow trout (Oncorhynchus mykiss) by  potential probiotic bacteria (Lactobacillus rhamnosus). Fish Shellfish Immunol. 15: 443-452.
Notash, Sh., Naeemi Kararoodi, M., Shahabzadeh, S.H., Fadaeifard, F. (2009) Study on the effect of different amount of probiotic protexin on weight gain, survival rate and FCR of rainbow trout. J Food Microbiol. 2: 33-40.
Ramos, M.A., Weber, B, Gonçalves, J.F, Santos, G.A., Rema, P., Ozório. R. (2013) Dietary probiotic supplementation modulated gut microbiota and improved growth of juvenile rainbow trout (Oncorhynchus mykiss). Comp Biochem Physiol., Part A Mol Integr Physiol. 166: 302-307.
Rungruangsak-Torrissen, K., Rustad, A., Sunde, J., Eiane, S.A., Jensen, H.B., Opstvedt, J., Nygrd, E., Samuelsen, T.A., Mundheim, H., Luzzana, U., Venturini, G. (2002) In vitro digestibility based on fish crude enzyme extract for prediction of feed quality in growth trials. J Sci Food Agric. 82: 644-654.
Salas-Leiton, E., Anguis, V., Martín-Antonio, B., Crespo, D.V., Planas, J., Infante, C., Cañavate, J.P., Manchado, M. (2010) Effects of stocking density and feed ration on growth and gene expression in the Senegalese sole (Solea senegalensis)  Potential effects on the immune response. Fish Shellfish Immunol. 28: 296-302.
Shyne Anand, P.S., Kohli, M.P.S., Dam, R., Sundaray,  J.K., Kumar, S., Sinha, A., Pailan, G.H., Sukham, M.K. (2013) Effect of dietary supplementation of periphyton on growth performance and digestive enzyme activities in Penaeus monodon. Aquaculture. 392-395: 59-68.
Skrodenytė-Arbačiauskienė, V., Sruoga, A., Butkauskas, D., Skrupskelis, K. (2008) Phylogenetic analysis of intestinal bacteria of freshwater salmon Salmo salar and sea trout  Salmo trutta trutta and diet. Fish Sci. 74: 1307-1314.
Soleimani, N., Hoseinifar, S.H., Merrifield, D.L., Barati, M., Abadi, Z.H. (2012) Dietary supplementation of fructooligosaccharide (FOS) improves the innate immune response, stress resistance, digestive enzyme activities and growthperformance of Caspian roach (Rutilus rutilus) fry. Fish Shellfish Immunol. 32: 316-321.
Sun, Y.Z., Yang, H.L., Ma, R.L., Lin, W.Y. (2011) Probiotic applications of two dominant gut Bacillus strains with antagonistic activity impoved the growth performance and immune responses of grouper Epinephelus coioides. Fish Shellfish Immunol. 29: 803-809.
Sun, Y.Z., Yang, H.L,. Ma, R.L., Song, K., Li, J.S. (2011) Effect of Lactococcus lactis and Enterococcus faecium on growth performance, digestive enzymes and immune response of grouper Epinephelus coioides. Aquac Nutr. 18: 281-289.
Suzer, C., Çoban, D., Kamaci, H.O., Saka, S., Firat, K., Otgucuoğlu, Ö., Küçüksari, H. (2008) Lactobacillus spp. bacteria as probiotics in gilthead sea bream (Sparus aurata, L.) larvae Effects on growth performance and digestive enzyme activities. Aquaculture. 280: 140-145.
Tovar, D.,  Zambonino, J., Cahu, C., Gatesoupe, F.J., Vázquez-Juárez, R., Lésel, R. (2002) Effect of live yeast incorporation in compound diet on digestive enzyme activity in sea bass (Dicentrarchus labrax) larvae. Aquaculture.  204: 113-123.  
Venkat, H.K., Sahu, N.P., Jain, K.K. (2004) Effect of feeding Lactobacillus-based probiotics on the gut microflora, growth and survival of postlarvae of Macrobrachium rosenbergii (de Man). Aquac Res. 35: 501-507.
Verschuere, L., Rombaut, G., Sorgeloos, P., Verstraete, W. (2000) Probiotic bacteria as biological control agents in aquaculture. Microbiol Mol Biol Rev. 64: 655-671.
Wang, Y.B. (2007) Effect of probiotics on growth performance and digestive enzyme activity of the shrimp Penaeus vannamei. Aquaculture. 269: 259-264.
Wang, Y.B., Tian, Z.Q., Yao, J.T.,  Li, W.F. (2008) Effect of probiotics, Enteroccus faecium, on tilapia (Oreochromis niloticus) growth performance and immune response. Aquaculture. 277: 203-207.
Wang, Y.B., Xu, Z. (2006) Effect of probiotics for common carp (Cyprinus carpio) based on growth performance and digestive enzyme activities. Anim Feed Sci Technol. 127: 283-292.
Worthington, C.C. (1991) Worthigton Enzyme Manual Related Biochemical. (3rd ed.) Freehold, New Jersy, USA.
Ziaei-Nejad, S., Rezaei, M.H., Takami, G.A., Lovett, D.L., Mirvaghefi, A.R., Shakouri, M. (2006) The effect of Bacillus spp. bacteria used as probiotics on digestive enzyme activity, survival and growth in the Indian Fenneropenaeus indicus. Aquaculture.  252: 516-524.