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Screening of Autochthonous Intestinal Microbiota as Candidate Probiotics Isolated from Four Freshwater Teleosts


Affiliations
1 Fisheries Laboratory, Department of Zoology, Visva-Bharati University, Santiniketan 731 235, India
2 Department of Zoology, University of Burdwan, Golap Bag, Burdwan 713 104, India
 

In this study altogether 109 autochthonous gut bacteria were screened from 4 fish species (Labeo rohita, Labeo bata, Catla catla and Puntius javanicus), of which 13 isolates displayed antagonism to 4 fish potent pathogens, namely Aeromonas salmonicida, Aeromonas hydrophila, Pseudomonas fluorescens and Aeromonas sobria. Eight promising isolates were further evaluated for extracellular enzyme production, non-hemolytic activity, bile tolerance and identified by 16S rRNA sequencing. Strains CCF7 (identified as Bacillus sp.), CCH9 and PJH1 (identified as two strains of Bacillus amyloliquefaciens) exhibited high score in antagonism assay and fulfilled other probiotic criteria, including safety aspects. However, application of these probiotics in aquaculture industries requires in vivo experiments and other information like immune modulating efficiency and binding ability on gut.

Keywords

Antagonism, Bacillus Species, Fish Gut Bacteria, Athogen, Probiotics.
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  • Subasinghe, R., Soto, D. and Jia, J., Global aquaculture and its role in sustainable development. Rev. Aquacult., 2009, 1, 2–9.

  • Cruz, P. M., Ibanez, A. L., Hermosillo, O. A. M. and Saad, H. C. R., Use of probiotics in aquaculture. ISRN Microbiol., 2012, 2012, 1–13.

  • Sugita, H., Shibuya, K., Shimooka, H. and Deguchi, Y., Antibacterial abilities of intestinal bacteria in freshwater cultured fish. Aquaculture, 1996, 145, 195–203.

  • Austin, B. and Austin, D. A., Characteristics of the diseases. In Bacterial Fish Pathogens: Disease in Farmed and Wild Fish, Springer-Praxis, Goldalming, UK, 2007, pp. 24–45.

  • Defoirdt, T., Sorgeloos, P. and Bossier, P., Alternatives to antibiotics for the control of bacterial disease in aquaculture. Curr. Opin. Microbiol., 2011, 14, 251–258.

  • Bøgwald, J. and Dalmo, R. A., Gastrointestinal pathogenesis in aquatic animals. In Aquaculture Nutrition: Gut Health, Probiotics and Prebiotics (eds Merrifield, D. L. and Ringø, E.), WileyBlackwell Publishing, Oxford, UK, 2014, pp. 53–74.

  • Cabello, F. C., Godfrey, H. P., Tomova, A., Ivanova, L., Dolz, H., Millanao, A. and Buschmann, A. H., Antimicrobial use in aquaculture re‐examined: its relevance to antimicrobial resistance and to animal and human health. Environ. Microbiol., 2013, 15, 1917–1942.

  • Pham, D. K., Chu, J., Do, N. T., Brose, F., Degand, G., Delahaut, P. and Wertheim, H. F., Monitoring antibiotic use and residue in freshwater aquaculture for domestic use in Vietnam. EcoHealth, 2015, 1–10.

  • Romero, J., Feijoó, C. G. and Navarrete, P., Antibiotics in aquaculture – use, abuse and alternatives. In Health and Environment in Aquaculture (ed. Carvalho, E.), InTech, Rijeka, Croatia, 2012, pp. 159–198.

  • Van Hai, N., Research findings from the use of probiotics in tilapia aquaculture: a review. Fish Shellfish Immunol., 2015, 45, 592–597.

  • Verschuere, L., Rombaut, G., Sorgeloos, P. and Verstraete, W., Probiotic bacteria as biological control agents in aquaculture. Microbiol. Mol. Biol. Rev., 2000, 64, 655–671.

  • Gatesoupe, F. J., The use of probiotics in aquaculture. Aquaculture, 1999, 180, 147–165.

  • Reddy, J., Probiotics in aquaculture: importance, influence and future perspectives. Int. J. Bioassays, 2015, 4, 3710–3718.

  • Joborn, A., Olsson, J. C., Westerdahl, A., Conway, P. L. and Kjelleberg, S., Colonization in the fish intestinal tract and production of inhibitory substances in intestinal mucus and faecal extracts by Carnobacterium sp. strain Kl. J. Fish Dis., 1997, 20, 383–392.

  • Sun, Y. Z., Yang, H. L., Ma, R. L. and Lin, W. Y., Probiotic applications of two dominant gut Bacillus strains with antagonistic activity improved the growth performance and immune responses of grouper Epinephelus coioides. Fish Shellfish Immunol., 2010, 29, 803–809.

  • Askarian, F., Kousha, A., Salma, W. and Ringø, E., The effect of lactic acid bacteria administration on growth, digestive enzyme activity and gut microbiota in Persian sturgeon (Acipenser persicus) and beluga (Huso huso) fry. Aquacult. Nutr., 2011, 17, 488–497.

  • Talpur, A. D., Memon, A. J., Khan, M. I., Ikhwanuddin, M., Daniel, M. M. D. and Abol-Munafi, A. B., Inhibition of pathogens by lactic acid bacteria and application as water additive multi isolates in early stages larviculture of P. pelagicus (Linnaeus, 1758). J. Anim. Plant Sci., 2012, 22, 54–64.

  • Ambas, I., Suriawan, A. and Fotedar, R., Immunological responses of customised probiotics-fed marron, Cherax tenuimanus (Smith 1912) when challenged with Vibrio mimicus. Fish Shellfish Immunol., 2013, 35, 262–270.

  • Mukherjee, A. and Ghosh, K., Antagonism against fish pathogens by cellular components and verification of probiotic properties in autochthonous bacteria isolated from the gut of an Indian major carp, Catla catla (Hamilton). Aquacult. Res., 2014, 47, 1–13.

  • Muthukumar, P. and Kandeepan, C., Isolation, identification and characterization of probiotic organisms from intestine of freshwater fishes. Int. J. Curr. Microbiol. Appl. Sci., 2015, 4, 607–616.

  • Bairagi, A., Ghosh, K., Sen, S. K. and Ray, A. K., Enzyme producing bacterial flora isolated from fish digestive tracts. Aquacult. Int., 2002, 10, 109–121.

  • Banerjee, G., Ray, A. K., Askarian, F. and Ringø, E., Characterisation and identification of enzyme-producing autochthonous bacteria from the gastrointestinal tract of two Indian air-breathing fish. Beneficial. Microb., 2013, 4, 277–284.

  • Ramkumar, R., Ravi, M., Jayaseelan, C., Rahuman, A. A., Anandhi, M., Rajthilak, C. and Perumal, P., Description of Providencia vermicola isolated from diseased Indian major carp, Labeo rohita (Hamilton, 1822). Aquaculture, 2014, 420, 193–197.

  • Dopazo, C. P., Lemos, M. L., Lodeiros, C., Bolinches, J., Barja, J. L. and Toranzo, A. E., Inhibitory activity of antibiotic producing marine bacteria against fish pathogens. J. Appl. Bacteriol., 1988, 65, 97–101.

  • Sangiliyandi, G. and Gunasekeran, P., Extracellular lipase producing Bacillus licheniformis from an oil mill refinery effluent. Indian J. Microbiol., 1996, 36, 109–110.

  • Bernfeld, P., Amylase [alpha] and [beta]. In Methods in Enzymology (eds Kolowick, S. P. and Kaplan, N. O.), Academic Press, New York, USA, 1955, pp. 149–150.

  • Walter, H. E., Methods of Enzymatic Analysis, Verlag Chemie, Weinheim, Germany, 1984, p. 238.

  • Denison, D. A. and Koehn, R. D., Cellulase activity of Poronia oedipus. Mycologia, 1977, 69, 592–601.

  • Bier, M., Lipases. In Methods in Enzymology (eds Kolowick, S. P. and Kaplan, N. O.), Academic Press, New York, USA, 1955, pp. 627–642.

  • Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Randall, R. J., Protein measurement with the folin phenol reagent. J. Biol. Chem., 1951, 93, 265–275.

  • Nikoskelainen, S., Salminen, S., Bylund, G. and Ouwehand, A., Characterization of the properties of human and dairy-derived probiotics for prevention of infectious diseases in fish. Appl. Environ. Microbiol., 2001, 67, 2430–2435.

  • Nayak, S. K. and Mukherjee, S. C., Screening of gastrointestinal bacteria of Indian major carps for a candidate probiotic species for aquaculture practices. Aquacult. Res., 2011, 42, 1034–1041.

  • Ringø, E., The ability of carnobacteria isolated from fish intestine to inhibit growth of fish pathogenic bacteria: a screening study. Aquacult. Res., 2008, 39, 171–180.

  • Vine, N. G., Leukes, W. D. and Kaiser, H., Probiotics in marine larviculture. FEMS Microbiol. Rev., 2006, 30, 404–427.

  • Ray, A. K., Roy, T., Mondal, S. and Ringø, E., Identification of gut-associated amylase, cellulase and protease-producing bacteria in three species of Indian major carps. Aquacult. Res., 2010, 41, 1462–1469.

  • Dutta, D. and Ghosh, K., Screening of extracellular enzymeproducing and pathogen inhibitory gut bacteria as putative probiotics in mrigal, Cirrhinus mrigala (Hamilton, 1822). Int. J. Fish. Aquat. Stud., 2015, 2, 310–318.

  • Thankappan, B., Ramesh, D., Ramkumar, S., Natarajaseenivasan, K. and Anbarasu, K., Characterization of Bacillus spp. from the gastrointestinal tract of Labeo rohita – towards to identify novel probiotics against fish pathogens. Appl. Biochem. Biotechnol., 2015, 175, 340–353.

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

  • Dutta, D., Banerjee, S., Mukherjee, A. and Ghosh, K., Selection and probiotic characterization of exoenzyme-producing bacteria isolated from the gut of Catla catla (Actinopterygii : Cypriniformes : Cyprinidae). Acta Ichthyol. Piscat., 2015, 45, 373–384.

  • Ramesh, D., Vinothkanna, A., Rai, A. K. and Vignesh, V. S., Isolation of potential probiotic Bacillus spp. and assessment of their subcellular components to induce immune responses in Labeo rohita against Aeromonas hydrophila. Fish Shellfish Immunol., 2015, 45, 268–276.

  • Ouwehand, A., Vankerckhoven, V., Goossens, H., Huys, G., Swings, J., Vancanneyt, M. and Lähteenmäki, A., The safety of probiotics in foods in Europe and its legislation. In Probiotics in Food Safety and Human Health (eds Goktepe, I., Juneja, V. K. and Ahmedna, M.), CRC Press, 2005, pp. 405–429.

  • Loh, J. Y., Lim, Y. Y., Harmin, S. A. and Ting, A. S. Y., In vitro assessment on intestinal microflora from commonly farmed fishes for control of the fish pathogen Edwardsiella tarda. Turk. J. Vet. Anim. Sci., 2014, 38, 257–263.

  • Askarian, F., Zhou, Z., Olsen, R. E., Sperstad, S. and Ringø, E., Culturable autochthonous gut bacteria in Atlantic salmon (Salmo salar L.) fed diets with or without chitin. Characterization by 16S rRNA gene sequencing, ability to produce enzymes and in vitro growth inhibition of four fish pathogens. Aquaculture, 2012, 326, 1–8.

  • Mondal, S., Roy, T. and Ray, A. K., Characterization and identification of enzyme‐producing bacteria isolated from the digestive tract of bata, Labeo bata. J. World Aquacult. Soc., 2010, 41, 369–377.

  • Cao, H., He, S., Wei, R., Diong, M. and Lu, L., Bacillus amyloliquefaciens G1: a potential antagonistic bacterium against eel-pathogenic Aeromonas hydrophila. J. Evid. Based Complement. Altern. Med., 2011, 2011, 1–7.


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  • Screening of Autochthonous Intestinal Microbiota as Candidate Probiotics Isolated from Four Freshwater Teleosts

Abstract Views: 388  |  PDF Views: 126

Authors

Ankita Nandi
Fisheries Laboratory, Department of Zoology, Visva-Bharati University, Santiniketan 731 235, India
Goutam Banerjee
Fisheries Laboratory, Department of Zoology, Visva-Bharati University, Santiniketan 731 235, India
Suhas Kumar Dan
Fisheries Laboratory, Department of Zoology, Visva-Bharati University, Santiniketan 731 235, India
Pinki Ghosh
Fisheries Laboratory, Department of Zoology, Visva-Bharati University, Santiniketan 731 235, India
Koushik Ghosh
Department of Zoology, University of Burdwan, Golap Bag, Burdwan 713 104, India
Arun Kumar Ray
Fisheries Laboratory, Department of Zoology, Visva-Bharati University, Santiniketan 731 235, India

Abstract


In this study altogether 109 autochthonous gut bacteria were screened from 4 fish species (Labeo rohita, Labeo bata, Catla catla and Puntius javanicus), of which 13 isolates displayed antagonism to 4 fish potent pathogens, namely Aeromonas salmonicida, Aeromonas hydrophila, Pseudomonas fluorescens and Aeromonas sobria. Eight promising isolates were further evaluated for extracellular enzyme production, non-hemolytic activity, bile tolerance and identified by 16S rRNA sequencing. Strains CCF7 (identified as Bacillus sp.), CCH9 and PJH1 (identified as two strains of Bacillus amyloliquefaciens) exhibited high score in antagonism assay and fulfilled other probiotic criteria, including safety aspects. However, application of these probiotics in aquaculture industries requires in vivo experiments and other information like immune modulating efficiency and binding ability on gut.

Keywords


Antagonism, Bacillus Species, Fish Gut Bacteria, Athogen, Probiotics.

References





DOI: https://doi.org/10.18520/cs%2Fv113%2Fi04%2F767-773