Distribution of extracellular enzyme-producing bacteria in the digestive tracts of 4 brackish water fish species

Occurrence and distribution of enzyme-producing bacteria in the proximal (PI), middle (MI), and distal (DI) segments of the gastrointestinal tracts of 4 brackish water teleosts (Scatophagus argus, Terapon jarbua, Mystus gulio, and Etroplus suratensis) have been investigated. Data were presented as log viable counts g-1 intestine (LVC). The heterotrophic bacterial population had the highest occurrence in the DI regions of all fish species studied except M. gulio. Proteolytic and amylolytic bacteria had the highest occurrence in the DI of M. gulio (LVC = 5.50 and 5.93, respectively), while cellulolytic and lipolytic populations exhibited highest occurrences in the DI regions of T. jarbua (LVC = 6.33) and S. argus (LVC = 5.78), respectively. Out of the 81 bacterial isolates, the most promising 3 isolates were determined through quantitative enzyme assay and studied through 16S rRNA gene sequence analysis for identification. Both the strains SA2.2 isolated from S. argus and TJ2.3 isolated from T. jarbua showed high similarity to different strains of Brevibacillus parabrevis, while another strain, MG4.2, isolated from M. gulio, was similar to Bacillus licheniformis. The NCBI GenBank accession numbers of the 16S rRNA gene sequences for isolates SA2.2, TJ2.3, and MG4.2 were KF377322, KF377324, and KF377323, respectively. The present study might offer scope for further research to evaluate prospects for application of the gut-associated extracellular enzyme-producing bacteria in brackish water aquaculture.

Distribution of extracellular enzyme-producing bacteria in the digestive tracts of 4 brackish water fish species

Occurrence and distribution of enzyme-producing bacteria in the proximal (PI), middle (MI), and distal (DI) segments of the gastrointestinal tracts of 4 brackish water teleosts (Scatophagus argus, Terapon jarbua, Mystus gulio, and Etroplus suratensis) have been investigated. Data were presented as log viable counts g-1 intestine (LVC). The heterotrophic bacterial population had the highest occurrence in the DI regions of all fish species studied except M. gulio. Proteolytic and amylolytic bacteria had the highest occurrence in the DI of M. gulio (LVC = 5.50 and 5.93, respectively), while cellulolytic and lipolytic populations exhibited highest occurrences in the DI regions of T. jarbua (LVC = 6.33) and S. argus (LVC = 5.78), respectively. Out of the 81 bacterial isolates, the most promising 3 isolates were determined through quantitative enzyme assay and studied through 16S rRNA gene sequence analysis for identification. Both the strains SA2.2 isolated from S. argus and TJ2.3 isolated from T. jarbua showed high similarity to different strains of Brevibacillus parabrevis, while another strain, MG4.2, isolated from M. gulio, was similar to Bacillus licheniformis. The NCBI GenBank accession numbers of the 16S rRNA gene sequences for isolates SA2.2, TJ2.3, and MG4.2 were KF377322, KF377324, and KF377323, respectively. The present study might offer scope for further research to evaluate prospects for application of the gut-associated extracellular enzyme-producing bacteria in brackish water aquaculture.

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  • Asfie, M., Yoshijima, T. and Sugita, H. 2003. Characterization of the goldfish fecal microflora by the fluorescent in situ hybridization method. Fisheries Sci. 69: 21–26.
  • Askarian, F., Kousha, A., Salma, W. and Ringİ, E. 2011. The effect of lactic acid bacteria administration on growth, digestive enzymes activity and gut microbiota in Persian sturgeon (Acipenser persicus) and beluga (Huso huso) fry. Aquacult. Nutr. 17(5): 488–497.
  • Askarian, F., Zhou, Z., Olsen, R.E., Sperstad, S. and Ringİ, E. 20 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 326–329: 1–8. Austin, B. 2002. The bacterial microflora of fish. Sci. World J. 2: 558–572.
  • Bairagi, A., Ghosh, K.S., Sen, S.K. and Ray, A.K. 2002. Enzyme producing bacterial flora isolated from fish digestive tracts. Aquacult. Int. 10: 109–121.
  • Bairagi, A., Sarkar Ghosh, K., Sen, S.K. and Ray, A.K. 2004. Evaluation of nutritive value of Leucaena leucocephala leaf meal inoculated with fish intestinal bacteria Bacillus subtilis and Bacillus circulans in formulated diets for rohu, Labeo rohita (Hamilton) fingerlings. Aquacult. Res. 35: 436–446.
  • Belchior, S.G. and Vacca, G. 2006. Fish protein hydrolysis by a psychrotrophic marine bacterium isolated from the gut of hake
  • (Merluccius hubbsi). Can. J. Microbiol. 52: 1266–1271.
  • Bernfeld, P. 1955. Amylase. In: Methods in Enzymology (Eds. S.P. Colowick and N.O. Kaplan). Academic Press, New York, NY, USA, pp. 149–158.
  • Beveridge, M.C.M., Sikdar, P.K., Frerichs, G.N. and Millar, S. 1991.
  • The ingestion of bacteria in suspension by the common carp Cyprinus carpio L. J. Fish Biol. 39: 825–831. Bier, M. 1955. Methods in Enzymology. Vol. I. Academic Press, New York, NY, USA, pp. 627–642.
  • Clements, K.D. 1997. Fermentation and gastrointestinal microorganisms in fishes. In: Gastrointestinal Microbiology.
  • Vol I. Gastrointestinal Ecosystems and Fermentations (Eds. R.I. Mackie and B.A. White). Chapman and Hall, New York, NY, USA, pp. 156–198. Creach, P.V. 1963. The proteolytic enzymes of poisons. In: Nutrition in Poecilothermes (Ed. CNRS). CNRS, Paris, France, pp. 375– 4
  • Dan, S.K. and Ray, A.K. 2013. Characterization and identification of phytase-producing bacteria isolated from the gastrointestinal tract of four freshwater teleosts. Ann. Microbiol. DOI: 10.1007/ s13213-013-0664-3.
  • Danulat, E. and Kausch, H. 1984. Chitinase activity in the digestive tract of cod, Gadus morhua (L.). J. Fish Biol. 24: 125–133.
  • Das, K.M. and Tripathi, S.D. 1991. Studies on the digestive enzymes of grass carp, Ctenopharyngodon idella (V). Aquaculture 92: 21–
  • De, D., Ghoshal, T.K. and Ananda Raja, R. 2012. Characterization of enzyme-producing bacteria isolated from the gut of Asian seabass, Lates calcarifer and milkfish, Chanos chanos and their application for nutrient enrichment of feed ingredients. Aquacult. Res. DOI: 10.1111/are.12099.
  • Denison, D.A. and Koehn, R.D. 1977. Cellulase activity of Poronia oedipus. Mycologia 69: 592–601.
  • Esakkiraj, P., Immanuel, G., Sowmya, S.M., Iyapparaj, P. and Palavesam, A. 2009. Evaluation of protease-producing ability of fish gut isolate Bacillus cereus for aqua feed. Food Bioprocess Technol. 2(4): 383–390.
  • Finegold, S.M., Sutter, V.L. and Mathisen, G.E. 1983. Normal indigenous intestinal flora. In: Human Intestinal Microflora in Health and Disease (Ed. D.J. Hentgens). Academic Press, New York, NY, USA, pp. 3–31.
  • Ghosh, K., Roy, M., Kar, N. and Ringİ, E. 2010. Gastrointestinal bacteria in rohu, Labeo rohita (Actinopterygii: Cypriniformes: Cyprinidae): scanning electron microscopy and bacteriological study. Acta Ichthyol. Piscatoria 40(2): 129–135.
  • Ghosh, K., Sen, S.K. and Ray, A.K. 2002a. Characterization of bacilli isolated from the gut of rohu, Labeo rohita, fingerlings and its significance in digestion. J. Appl. Aquacult. 12(3): 33–42.
  • Ghosh K., Sen, S.K. and Ray, A.K. 2002b. Growth and survival of rohu, Labeo rohita (Hamilton) spawn fed diets supplemented with fish intestinal microflora. Acta Ichthyol. Piscatoria 32(1): 83–
  • Ghosh, K., Sen, S.K. and Ray, A.K. 2003. Supplementation of an isolated fish gut bacterium, Bacillus circulans, in formulated diets for rohu, Labeo rohita, fingerlings. Israeli J. Aquacult. 55: 13–
  • Ghosh, K., Sen, S.K. and Ray, A.K. 2004a. Growth and survival of rohu, Labeo rohita (Hamilton, 1822) spawn fed diets fermented with intestinal bacterium, Bacillus circulans. Acta Ichthyol. Piscatoria 34(2): 155–165.
  • Ghosh, K., Sen, S.K. and Ray, A.K. 2004b. Supplementation of Lactobacillus acidophilus in compound diets for Labeo rohita fingerlings. Indian J. Fish. 51(4): 521–526.
  • Horsley, R.W. 1977. A review of the bacterial flora of teleosts and elasmobranches, including methods for its analysis. J. Fish Biol. 10: 529–553.
  • Itoi, S., Okamura, T., Koyama, Y. and Sugita, H. 2006. Chitinolytic bacteria in the intestinal tract of Japanese coastal fishes. Can. J. Microbiol. 52: 1158–1163.
  • Jacob, M.B. and Gerstein, M.J. 1960. Handbook of Microbiology. D. Van Nostrand Co. Inc., Princeton, NJ, USA.
  • Kar, N., Roy, R.N., Sen, S.K. and Ghosh, K. 2008. Isolation and characterization of extracellular enzyme-producing bacilli in the digestive tracts of rohu, Labeo rohita (Hamilton) and murrel, Channa punctatus (Bloch). Asian Fish. Sci. 21(4): 421–434.
  • Khan, A. and Ghosh, K. 2012. Characterization and identification of gut-associated phytase-producing bacteria in some freshwater fish cultured in ponds. Acta Ichthyol. Piscatoria 42(1): 37–45.
  • Khan, A. and Ghosh, K. 2013. Evaluation of phytase production by fish gut bacterium, Bacillus subtilis for processing of Ipomea aquatica leaves as probable aquafeed ingredient. J. Aquat. Food Prod. Technol. 22(5): 508–519.
  • Khan, A., Mandal, S., Samanta, D., Chatterjee, S. and Ghosh, K. 2011.
  • Phytase-producing Rhodococcus sp. (MTCC 9508) from fish gut: a preliminary study. Proc. Zool. Soc. 64: 29–34.
  • Kinnear P.R. and Gray, C.D. 2000. SPSS for Windows Made Simple.
  • Release 10. Psychology Press, Sussex, UK. Lane, D.J. 1991. 16S/23S rRNA sequencing. In: Nucleic Acid Techniques in Bacterial Systematics (Eds. E. Stackebrandt and M. Goodfellow). Wiley, Chichester, UK, pp. 130–141.
  • Li, X., Chi, Z., Liu, Z., Yan, K. and Li, H. 2008. Phytase production by a marine yeast Kodamea ohmeri BG3. Applied Biochem. Biotechnol. 149: 183–193.
  • Li, X.Y., Liu, Z.Q. and Chi, Z.M. 2008. Production of phytase by a marine yeast Kodamaea ohmeri BG3 in an oats medium: optimization by response surface methodology. Bioresour. Technol. 99: 6386–6390.
  • Luczkovich, J.J. and Stellwag, E.J. 1993. Isolation of cellulolytic microbes from the intestinal tract of pinfish, Lagodon rhomboids: size-related changes in diet and microbial abundance. Mar. Biol. 116: 381–388.
  • MacDonald, N.L., Stark, J.R. and Austin, B. 1986. Bacterial microflora in the gastrointestinal tract of Dover sole (Solea solea L.), with emphasis on the possible role of bacteria in the nutrition of the host. FEMS Microbiol. Lett. 35: 107–111.
  • Mandal, S. and Ghosh, K. 2013. Isolation of tannase-producing microbiota from the gastrointestinal tracts of some freshwater fish. J. Appl. Ichthyol. 29: 145–153.
  • Mondal, S., Roy, T. and Ray, A.K. 2010. Characterization and identification of enzyme-producing bacteria isolated from the digestive tract of bata, Labeo bata. J. World Aquacult. Soc. 41: 369–377.
  • Mondal, S., Roy, T., Sen, S.K. and Ray, A.K. 2008. Distribution of enzyme-producing bacteria in the digestive tracts of some freshwater fish. Acta Ichthyol. Piscatoria 38: 1–8.
  • Rahmatullah, S.M. and Beveridge, M.C.M. 1993. Ingestion of bacteria in suspension by Indian major carps (Catla catla, Labeo rohita) and Chinese carps (Hypophthalmichthys molitrix, Aristichthys nobilis). Hydrobiologia 264: 79–84.
  • Ramachandran, S. and Ray, A.K. 2007. Nutritional evaluation of fermented black gram (Phaseolus mungo) seed meal in compound diets for rohu, Labeo rohita (Hamilton), fingerlings. J. Appl. Ichthyol. 23: 74–79.
  • Ramachandran, S., Bairagi, A. and Ray, A.K. 2005. Improvement of nutritive value of grass pea (Lathyrus sativus) seed meal in the formulated diets for rohu, Labeo rohita (Hamilton) fingerlings after fermentation with a fish gut bacterium. Bioresour. Technol. 96: 1465–1472.
  • Rani, S., Garg, S.K., Sabhlok, V.P. and Bhatnagar, A. 2004. Intestinal enzyme activity and enzyme-producing microbial flora in relation to feeding behaviour in some brackish water teleosts. J. Aquacult. 12: 55–68.
  • Ray, A.K., Ghosh, K. and Ringİ, E. 2012. Enzyme-producing bacteria isolated from fish gut: a review. Aquacult. Nutr. 18: 465–492.
  • Ray, A.K., Roy, T., Mondal, S. and Ringİ, E. 2010. Identification of gut-associated amylase, cellulase, and protease-producing bacteria in three species of Indian major carps. Aquacult. Res. 41:1462–1469.
  • Ringİ, E. 1993. The effect of chromic oxide (Cr 2 O 3 ) on aerobic bacterial populations associated with the epithelial mucosa of Arctic charr (Salvelinus alpinus L.). Can. J. Microbiol. 39: 1169–1173.
  • Ringİ, E. and Birkbeck, T.H. 1999. Intestinal microflora of fish larvae and fry. Aquacult. Res. 30: 73–93.
  • Ringİ, E., Olsen, R.E., Gifstad, T.O., Dalmo, R.A., Amlund, H., Hemre, G.I. and Bakke, A.M. 2010. Prebiotics in aquaculture: a review. Aquacult. Nutr. 16: 117–136.
  • Ringİ, E., Olsen, R.E., Mayhew, T.M. and Myklebust, R. 2003. Electron microscopy of the intestinal microflora of fish. Aquaculture 227: 395–415.
  • Roy, T., Mondal, S. and Ray, A.K. 2009. Phytase-producing bacteria in the digestive tracts of some freshwater fish. Aquacult. Res. 40: 344–353.
  • Saha, A. and Ray, A.K. 1998. Cellulase activity in rohu fingerlings. Aquacult. Int. 6: 281–291.
  • Saha, S. and Ray, A.K. 2011. Evaluation of nutritive value of water hyacinth (Eichhornia crassipes) leaf meal in compound diets for rohu, Labeo rohita (Hamilton, 1822) fingerlings after fermentation with two bacterial strains isolated from fish gut. Turkish J. Fish. Aqua. Sci. 11: 199–209.
  • Saha, S., Roy, R.N., Sen, S.K. and Ray, A.K. 2006. Characterization of cellulase-producing bacteria from the digestive tract of tilapia, Oreochromis mossambica (P) and grass carp, Ctenopharyngodon idella (V). Aquacult. Res. 37: 380–388.
  • Sakata, T. 1990. Microflora in the digestive tract of fish and shellfish.
  • In: Microbiology in Poecilotherms (Ed. R. Lesel). Elsevier, Amsterdam, the Netherlands, pp. 171–176. Sangiliyandi, G. and Gunasekaran, P. 1996. Extracellular lipaseproducing Bacillus licheniformis from an oil mill refinery effluent. Indian J. Microbiol. 36: 109–110.
  • Shcherbina, M.A. and Kazlawlene, O.P. 1971. The reaction of the medium and the rate of absorption of nutrients in the intestine of carp. J. Ichthyol. 11: 81–85.
  • Silva, F.C.P., Brito, M.F.G., Brito, L.M. and Nicoli, J.R. 2005. Composition and antagonistic activity of the indigenous intestinal microbiota of Prochilodus argenteus Agassiz. J. Fish Biol. 67: 1686–1698.
  • Sivasubramanian, K., Ravichandran, S. and Kavitha, R. 2012. Isolation and characterization of gut microbiota from some estuarine fishes. Marine Sci. 2(2): 1–6.
  • Smith, L.S. 1989. Digestive functions in teleost fishes. In: Fish Nutrition (Ed. J.E. Halver). Academic Press, San Diego, CA, USA, pp. 331–42.
  • Stavric, S. and Kornegay, E.T. 1995. Microbial probiotics for pigs and poultry. In: Biotechnology in Animal Feeds and Animal
  • Feeding (Eds. R.J. Wallace and A. Chuson). Wiley, New York, NY, USA, pp. 205–231. Stickney, R.R. and Shumway, S.E. 1974. Occurrence of cellulase activity in the stomachs of fishes. J. Fish Biol. 6: 779–790.
  • Sugita, H., Shibuya, K., Harada, H. and Deguchi, Y. 1997. Antibacterial abilities of intestinal microflora of the river fish. Fisheries Sci. 63: 378–383.
  • Teather, R.M. and Wood, P.J. 1982. Use of Congo red-polysaccharide interactions in enumeration and characterization of cellulolytic bacteria from the bovine rumen. Appl. Environ. Microbiol. 43: 777–780.
  • Trust, T.J. and Sparrow, R.A. 1974. The bacterial flora in the alimentary tract of fresh water salmonid fishes. Can. J. Microbiol. 20: 1219–1228.
  • Walter, H.E. 1984. Methods of Enzymatic Analysis. Verlag Chemie, Weinheim, Germany.
  • Zar, J.E. 1974. Biostatistical Analysis. Prentice Hall, NJ, USA.
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