Characterization, screening, and application of bacteria with probiotic properties isolated from the gut of Labeo calbasu (Hamilton)

  • 1 Department of Zoology, 136119, Kurukshetra, India


Labeo calbasu is not reared in aquaculture because of the unavailability of proper feed and the risk of infection. The ability of probiotics to promote growth and immunological defense barriers in the gut has led to the use of autochthonous probiotic bacteria. Hence, the present study was conducted to screen potential probiotic bacteria from the gut of L. calbasu and to supplement the same to study the effect on growth and immunity. Strains with probiotic properties were assayed for exogenous enzyme production and identified based on phylogenetic analysis and nucleotide homology. Four of nine isolates showed promising antibacterial activity against Aeromonas hydrophila. The strains survived in relatively low pH and high bile concentrations and were investigated for hydrophobicity, autoaggregation, and coaggregation. Based on the positive results of this study, the LC1 and LC4 strains (analysed with 16S rRNA gene sequencing and showing similarity with Aneurinibacillus aneurinilyticus and Stenotrophomonas sp.) were further incorporated into diets to explore their probiotic effects on the growth performance and immunity of the fish. Administering the probiotics elevated growth parameters and benefitted the immune system by increasing the phagocytic index and decreasing the mortality rate with exposure to A. hydrophila.

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  • American Public Health Association 1998 – Standard methods for the examination of water and waste water – 20th edn. American Public Health Association, New York.

  • Bairagi A., Sarkarghosh K., Sen S.K., Ray A.K. 2002 – Duck-weed (Lemna polyrrhiza) leaf meal as source of feedstuff in formulated diets for rohu (Labeo rohita) fingerlings after formulation with a fish intestinal bacterium – Bioresour. Technol. 85: 17-23.

  • Balcazar J.L., de Blas I., Ruiz-Zarzuela I., Cunningham D., Vendrell D., Muzquiz J.L. 2006 – The role of probiotics in aquaculture – Vet. Microbiol. 114: 173-186.

  • Balcazar J.L., Blas D., Zarzuela I.R., Vendrell D., Girones O., Muzquiz J.L. 2007a – Sequencing of variable regions of the 16S rRNA gene for identification of lactic acid bacteria isolated from the intestinal microbiota of healthy salmonids – Comp. Immunol. Microbiol. Infect. Dis. 30: 111-118.

  • Balcazar J.L., de Blas I., Ruiz-Zarzuela I., Vendrell D., Girones O., Muzquiz J.L. 2007b – Enhancement of the immune response and protection induced by probiotic lactic acid bacteria against furunculosis in rainbow trout (Oncorhynchus mykiss) – FEMS Immunol. Med. Mic. 51: 185-193.

  • Bandyopadhyay P. 2004 – Improvement of growth, nutritional quality and immunity of Indian major carps through probiotic supplement – Ph.D. thesis, Vidyasagar University, Midnapore-721102, West Bengal. India.

  • Barry T., Powell R., Gannon F.A. 1990 – A general method for the development of DNA probes for microorganisms – Nat. Biotechnol. 8: 233-236

  • Bergey’s Manual of Systematics of Archaea and Bacteria 2015 – Bergey’s Manual Trust – John Wiley and Sons.

  • Berwick V., Cheek L., Ball J. 2004 – Statistics review 12: Survival analysis – Crit. Care 8: 389-394.

  • Bhatnagar A., Dhillon O. 2017 – Evaluation of optimum protein requirement and cost effective eco-friendly source for Labeo calbasu (Hamilton, 1822) – J. Fish. Aquat. Sci. 12: 273-283.

  • Bhatnagar A., Lamba R. 2015 – Antimicrobial ability and growth promoting effects of feed supplemented with probiotic bacterium isolated from gut microflora of Cirrhinus mrigala – J. Integr. Agric. 14: 583-592.

  • Bhatnagar A., Lamba R. 2017 – Molecular characterization and dosage application of autochthonous potential probiotic bacteria in Cirrhinus mrigala – J Fisheries 11(2): 46-56.

  • Bhatnagar A., Raparia S. 2014 – Optimum dietary inclusion level of Bacillus coagulans for growth and digestibility improvement for Catla catla (Hamilton) – Int. J. Curr. Res. Rev. 6(7): 1–10.

  • Bhatnagar A., Raparia S., Kumari S. 2012 – Isolation and Influence of Bacillus coagulans CC1 on growth performance and digestive enzyme activities of Catla catla –J. Nat. Sci. Sustainable Technol. 6: 237-253

  • Carnevali O., de Vivo L., Sulpizio R., Gioacchini G., Olivotto I., Silvi S., Cresci A. 2006 – Growth improvement by probiotic in European sea brass juveniles (Dicentrachus labrax, L.), with particular attention to IGF-1, myostatin and cortiosol gene expression – Aquaculture 258: 430-438

  • Conway P.L., Gorbach S.L., Goldin B.R. 1987 – Survival of lactic acid bacteria in the human stomach and adhesion to intestinal cells – J. Dairy Sci. 70: 1-12.

  • Das D., Chatterjee S.N., Dangar T.K. 2016 – Characterization and mosquitocidal potential of the soil bacteria Aneurinibacillus aneurinilyticus isolated from Burdwan, West Bengal, India – Proc. Natl. Acad. Sci. India Sect. B Biol. Sci. 86: 707-713.

  • Del Re B., Sgorbati D., Miglioli M., Palenzona D. 2000 – Adhesion, autoaggregation and hydrophobicity of 13 strains of Bifidobacterium longum – Lett. Appl. Microbiol. 31: 438-442.

  • Duncan D.B. 1955 – Multiple range and Multiple F Tests – Biometrics 11: 1-42.

  • Ellis A.E. 1988 – Fish Vaccination – Academic Press, London, 84 p.

  • Garg S.K., Bhatnagar A., Kalla A., Johal M.S. 2002 – Experimental Ichthyology – CBS Publications and Distributors, New Delhi.

  • Ghosh K., Sen S.K., Ray A.K. 2002 – Characterization of Bacilli isolated from gut of rohu, Labeo rohita, fingerlings and its significance in digestion – J. Appl. Aquacult. 12: 33-42.

  • Ghosh S., Sinha A., Sahu C. 2007 – Isolation of putative probionts from the intestines of Indian major carps – Isr. J. Aquacult. – Bamidgeh 59: 127-132.

  • Gilliland S.E., Staley T.E., Bush L.J. 1984 – Importance of bile tolerance of Lactobacillus acidophilus used as a dietary adjunct – J. Dairy Sci. 67: 3045-3051.

  • Giri S.S., Sukumaran V., Oviya M. 2013 – Potential probiotic Lactobacillus plantarum VSG-3 improves the growth, immunity, and disease resistance of tropical freshwater fish, Labeo rohita – Fish Shellfish Immunol. 34: 660-666.

  • Handley P.S., Harty D.W.S., Wyatt J.E., Brown C.R., Doran J.P., Gibbs A.C.C. 1987 – A comparison of the adhesion, coaggregation and cell-surface hydrophobicity properties of fibrillar and fimbriate strains of Streptococcus salivarius – J Gen. Microbiol. 133: 3207-3217.

  • Holt G.J., Krieg N.R., Sneath P.H.A., Staley J.T., Williams S.T. 1994 – Bergey’s manual of determinative bacteriology – 9th edn. Philadelphia, PA: Lippincott Williams and Wilkins.

  • Hossain M.M., Uddin M.N., Islam M.N., Chakraborty S.C., Kamal M. 1999 – Study on the intestinal bacteria of Labeo rohita (Ham.) – Bangladesh J. Fish. Res. 3: 63-66.

  • Kos B., Šuškoviæ J., Vukoviæ S., Šimpraga M., Frece J., Matošiæ S. 2003 – Adhesion and aggregation ability of probiotic strain Lactobacillus acidophilus M92 – J. Appl. Microbiol. 94: 981-987

  • Lyon W.J., Glatz B.A. 1993 – Isolation and purification of propionicin PLG-1, a bacteriocin produced by a strain of Propiobacterium theonii – Appl. Environ. Microbiol. 59: 83-87.

  • Mohapatra S., Chakraborty T., Prusty A.K., Das P., Paniprasad K., Mohanta K.N. 2012 – Use of different microbial probiotics in the diet of rohu, Labeo rohita fingerlings: effects on growth, nutrient digestibility and retention, digestive enzyme activities and intestinal microflora – Aquac. Nutr. 18: 1-11.

  • Nandi A., Dan S.K., Banerjee G., Ghosh P., Ghosh K., Ringo O., Ray A.K. 2017 – Probiotic potential of autochthonous bacteria isolated from the gastrointestinal tract of four freshwater teleosts – Probiotics Antimicro Prot. 9: 12-21.

  • Nayak S.K. 2010 – Role of gastrointestinal microbiota in fish – Aquacult. Res. 41: 1553-1726.

  • Nayak S.K., Mukherjee S.C. 2011 – Screening of gastrointestinal bacteria of Indian major carps – Aquacult. Res. 42: 1034-1041.

  • O’Sullivan D.J. 2001 – Screening of intestinal microflora for effective probiotic bacteria – J. Agric. Food Chem. 49: 1751-1760.

  • Park K.H., Jeong H.D. 1996 – Enhanced resistance against Edwardsiella tarda infection in tilapia (Oreochromis niloticus) by administration of protein-bound polysaccharide – Aquaculture 141: 135-143.

  • Pennacchia C., Ercolini D., Blaiotta G., Pepe O., Mauriello G., Villani F. 2004 – Selection of Lactobacillus strains from fermented sausages for their potential use as probiotics – Meat Sci. 67: 309-317.

  • Rani S., Garg S.K., Sabhlok V.P., Bhatnagar A. 2004 – Intestinal enzyme activity and enzyme producing microbial flora in relation to feeding behavior in some brackish water teleosts – J. Aquacult. 12: 55-68.

  • Ray A.K., Ghosh K., Ringo E. 2012 – Enzyme-producing bacteria isolated from fish gut – A review – Aquac. Nutr. 18: 465-492.

  • Rojas M., Conway P.L. 1996 – Colonization by lactobacilli of piglet small intestinal mucus – J. Appl. Bacteriol. 81: 474-480.

  • Sahoo P.K., Mukherjee S.C., Sahoo S.K. 1998 – Aeromonas hydrophila versus Edwardsiella tarda: a pathoanatomical study in Clarias batrachus – J. Aquacult. 6: 57-66.

  • Salvadori L., Gioia D.D., Fava F., Barberio C. 2006 – Degradation of low-ethoxylated nonylphenols by a Stenotrophomonas strain and development of new phylogenetic probes for Stenotrophomonas spp detection – Curr. Microbiol. 52: 13-20.

  • Sansawat A., Thirabunyanon M. 2009 – Anti-Aeromonas hydrophila activity and characterisation of novel probiotic strains of Bacillus subtilis isolated from the gastrointestinal tract of giant freshwater prawns – Maejo. Int. J. Sci. Technol. 3: 77-87.

  • Savage D.C. 1992 – Growth phase cellular hydrophobicity and adhesion in vitro of Lactobacilli colonizing the keratinizing gastric epithelium in the mouse – Appl. Environ. Microbiol. 58: 1992-1995.

  • Siwicki A.K., Anderson D.P., Rumsey G.L. 1994 – Dietary intake of immunostimulants by rainbow trout affects non-specific immunity and protection against furuncuclosis – Vet. Immunol. Immunopathol. 41: 125-139.

  • Vine N.G., Leukes W.D., Kaiser H., Daya S., Baxter J., Hecht T. 2004 – Competition for attachment of aquaculture candidate probiotic and pathogenic bacteria on fish intestinal mucus – J. Fish. Dis. 27: 319-326.

  • Zhu H., Qu F., Zhu L.H. 1993 – Isolation of genomic DNAs from plants, fungi and bacteria using benzyl chloride – Nucleic Acids Res. 21: 5279-5280.


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