In order to study the diversity of putative lactic acid bacteria (LAB) in freshwater fish, 76 strains of LAB were isolated from intestines and identified by phenotypic tests and 16S rDNA gene sequencing. Phenotypic characterization of the isolates allowed the identification of 18 clusters at 78% similarity level by Hierarchical cluster analysis. Functional evenness index (E value) a measure of phenotypic diversity, was found to be quite high (0.7 approximately) in most of the samples. 16S rDNA gene sequencing identified the isolates as different strains of Lactobacillus plantarum, Lactobacillus pentosus, Lactobacillus fermentum, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus brevis, Lactobacillus reuteri, Lactobacillus salivarius, Pediococcus pentosaceus, Pediococcus acidilactici, Weissella paramesenteroides, Weissella cibaria, Enterococcus faecium and Enterococcus durans. Lactobacillus plantarum was found to be the dominating strain and LAB occurred more frequently in hindgut. Most of the strains showed good survival in acid and bile tolerance tests and antimicrobial activity against fish pathogen Aeromonas hydrophila. Different bacteriocin producing genes were detected in several strains by PCR. Selective strains with probiotic attributes could be potential candidates for freshwater aquaculture practices
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Abed, T.A. 2013. Evaluation of methods for the extraction and purification of DNA of cultured Lactobacillus colony isolated from dairy products. International Journal of Applied Microbiology and Biotechnology Research, 1: 20-25.
Allameh, S.K., Yusoff, F.M., Daud, H.M., Ringo, E., Ideris, A. and Saad. C.R. 2013. Characterization of a probiotic Lactobacillus fermentum isolated from snakehead, Channa striatus, stomach. Journal of the World Aquaculture Society, 44: 835-844. doi: 10.1111/jwas.12075
Ayo Olalusi, C.I., Adeiga, A., Edah, B. and Akintunde, G.B. 2012. Isolation, culture and identification of Lactobacillus species from the gut of Sphyraena sphyraena, Chrysisthysnigro digitatus and Clarias gariepinus for use as probiotics in fish feeds. Journal of Food Processing and Technology, 3: 135. doi: 10.4172/2157-7110.S1.009
Balcazar, J.L, de Blas, I., Ruiz-Zarzuela, I., Vendrell, D., Girones, O. and Muzquiz, J.L. 2007. Enhancementof the immune response and protection induced by probiotic lactic acid bacteria against furunculosis in rainbow trout (Oncorhynchus mykiss ). FEMS Immunology and Medical Microbiology, 51: 185- 193. doi: 10.1111/j.1574-695X.2007.00294.x
Balcazar, J.L., Vendrell, D., de Blas, I., Ruiz-Zarzuela, I., Muzquiz, J.L. and Girones, O. 2008. Characterization of probiotic properties of lactic acid bacteria isolated from intestinal microbiota of fish. Aquaculture, 278: 188-191. doi: 10.1016/j.aquaculture.2008.03.014
Barrett, E., Hayes, M., O Connor, P., Gardiner, G., Fitzgerald, G.F., Stanton, C., Paul Ross, R. and Hill. C. 2007. Salivaricin P: one of a family of two component anti-listerial bacteriocins produced by intestinal isolates of Lactobacillus salivarius. Applied and Environmental Microbiology, 73: 3719-3723. doi: 10.1128/AEM.00666-06
Beck, B.R., Kim, D., Jeon, J., Lee, S.M., Kim, H.K., Kim, O.J., Lee, J.I., Suh, B.S., Do, H.K., Lee, K.H., Holzapfel, W.H., Hwang, J.Y., Kwon, M.G. and Song, S.K. 2015. The effects of combined dietary probiotics Lactococcus lactis BFE920 and Lactobacillus plantarum FGL0001 on innate immunity and disease resistance in olive flounder (Paralicthys olivaceus). Fish and Shellfish Immunology, 42: 177-183. doi: 10.1016/j.fsi.2014.10.035
Belhadj, H., Harzallah, D., Bouamra, D., Khennouf, S., Dahamna, S. and Ghadbane, M. 2014. Phenotypic and genotypic characterization of some lactic acid bacteria isolated from bee pollen: a preliminary study. Bioscience Microbiota Food and Health, 33: 11-23. doi: 10.12938/bmfh.33.11
Ben Omar, N., Abriouel, H., Keleke, S., Sanchez Valenzuela, A., Martinez-Canamero, M., Lucas Lopez, R., Ortega, E and Galvez. A. 2008.
Bacteriocin-producing Lactobacillus strains isolated from poto poto, a Congolese fermented maize product, and genetic fingerprinting of their plantaricin operons. International Journal of Food Microbiology, 127: 18-25. doi: 10.1016/j.ijfoodmicro.2008.05.037
Boukhemis, M., Djeghri Hocine, B., Tahar, A. and Amrane. A. 2009. Phenotypic characterization of Lactobacillus strains isolated from different biotopes. African Journal of Biotechnology, 8: 5011-5020. doi: 10.5897/AJB2009.000-9439
Buntin, N., Chanthachum, S. and Hongpattarakere, T. 2008. Screening of lactic acid bacteria from gastrointestinal tracts of marine fish for their potential use as probiotics. Songklanakarin Jounal of Science and Technology, 30: 141-148.
Butprom, S., Phumkhachorn, P. and Rattanachaikunsopon, P. 2013. Effect of Lactobacillus plantarum C014 on innate immune response and disease resistance against Aeromonas hydrophila in hybrid catfish. The Scientific World Journal, doi: 10.1155/2013/392523
Cadieux, P., Wind, A., Sommer, P., Schaefer, L., Crowley, K., Britton, R.A. and Reid, G. 2008. Evaluation of reuterin production in urogenital probiotic Lactobacillus reuteri RC-14. Applied and Environmental Microbiology, 74:4645-4649. doi: 10.1128/AEM.00139-08
Carnevali, O., de Vivo, L., Sulpizio, R., Gioacchini, G., Olivotto, I., Silvi, S. and Cresc, A. 2006. Growth improvement by probiotic in European sea bass juveniles (Dicentrarchus labrax, L.), with particular attention to IGF-1, myostatin and cortisol gene expression. Aquaculture, 258: 430-438. doi:10.1016/j.aquaculture.2006.04.025
Ceapa, C., Lambert, J., van Limpt, K., Wels, M., Smokvina, T., Knol, J. and Kleerebezem, M. 2015. Correlation of Lactobacillus rhamnosus genotypes and carbohydrate utilization signatures determined by phenotype profiling. Applied and Environmental Microbiology, 81: 5458-5470.
Dash, G., Raman, R.P., Prasad, K.P., Makesh, M., Pradeep, M.A. and Sen, S. 2014. Evaluation of Lactobacillus plantarum as feed supplement on host associated microflora, growth, feed efficiency, carcass biochemical composition and immune response of giant freshwater prawn, Macrobrachium rosenbergii (de Man, 1879). Aquaculture, 432: 225-236. doi: 10.1016/j.aquaculture.2014.05.011 de Man, J.C., Rogosa, M. and Sharpe, M.E. 1960. A medium for the cultivation of lactobacilli. Journal of Applied Bacteriology, 23: 130-135. doi: 10.1111/j.1365-2672.1960.tb00188
Denev, S., Staykov, Y., Moutafchieva, R. and Beev, G. 2009. Microbial ecology of the gastrointestinal tract of fish and the potential application of probiotics and prebiotics in finfish aquaculture. International Aquatic Research, 1: 1-29.
Di Cagno, R., Minervini, G., Sgarbi, E., Lazzi, C., Bernini, V., Neviani, E. and Gobbetti, M. 2010. Comparison of phenotypic (Biolog System) and genotypic (random amplified polymorphic DNA-polymerase chain reaction, RAPD-PCR, and amplified fragment length polymorphism, AFLP) methods for typing Lactobacillus plantarum isolates from raw vegetables and fruits. International Journal of Food Microbiology, 143: 246-253. doi: 10.1016/j.ijfoodmicro.2010.08.018
Diep, D. B., Havarstein, L.S. and Nes, I.F. 1996. Characterization of the locus responsible for the bacteriocin production in Lactobacillus plantarum C11. Journal of Bacteriology, 178: 4472-4483. du Toit, M., Franz, C.M., Dicks, L.M. and Holzapfel, W.H. 2000. Preliminary characterization of bacteriocins produced by Enterococcus faecium and Enterococcus faecalis isolated from pig faeces. Journal of Applied Microbiology, 88: 482-494. doi: 10.1046/j.1365- 2672.2000.00986.x
Duangjitcharoen, Y., Kantachote, D., Prasitpuripreecha, C., Peerajan, S. and Chaiyasut. C. 2014. Selection and characterization of probiotic lactic acid bacteria with heterocyclic amine binding and nitrosamine degradation properties. Journal of Applied Pharmaceutical Science, 4: 014-023. doi: 10.7324/JAPS.2014.40703
Ghosh, S., Ringo, E., Selvam, A.D.G., Rahiman, K.M.M., Sathyan, N., Nifty, J. and Hatha, A.A.M. 2014. Gut associated lactic acid bacteria isolated from the estuarine fish Mugil cephalus: molecular diversity and antibacterial activities against pathogens. International Journal of Aquaculture, 4: 1-11. doi: 10.5376/ija.2014.04.0001
Giri, S.S., Sukumaran, V., Sen, S.S. and Jena, P.K. 2014. Effects of dietary supplementation of potential probiotic Bacillus subtilis VSG1singularly or in combination with Lactobacillus plantarum VSG3 or/and Pseudomonas aeruginosa VSG2 on the growth, immunity and disease resistance of Labeo rohita. Aquaculture Nutrition, 20: 163-171. doi: 10.1111/anu.12062
Hagi, T. and Hoshino, T. 2009. Screening and characterization of potential probiotic lactic acid bacteria from cultured common carp intestine. Bioscience Biotechnology and Biochemistry, 73: 1479-1483. doi: 10.1271/bbb. 80746
Hammes, W. P., Weiss, N. and Holzapfel, W.H. 1992. The genera Lactobacillus and Carnobacterium. In: A. Balows, H.G. Truper, M. Dworkn, W. Harder and K.H. Schleifer (Ed.), Prokaryotes, a Handbook for the biology of bacteria: Ecophysiology, Isolation, Identification, Application, vol. 2. Springer-Verlag, New York, 1508-1534.
Hickling, C. F. 1966. On the feeding process in white Amur, Ctenopharyngodon idella. Journal of Zoology, 148: 408-419. doi: 10.1111/j.1469- 7998.1966.tb02960.x
Huidrom, S., Deka, M. and Deblakshmi, R. 2012. Identification of lactic acid bacteria from the faeces of infant. Recent Research in Science and Technology, 4: 54-56.
Hurtado. A., Ben Othman, N., Chammem, N., Hamdi, M., Ferrer, S., Reguant, C., Bordons, A. and Rozes, N. 2011. Characterization of Lactobacillus isolates from fermented olives and their bacteriocin gene profiles. Food Microbiology, 28: 1514-1518. doi:10.1016/j.fm.2011.07.010
Jankauskiene, R. 2002. Defence mechanisms in fish: frequency of the genus Lactobacillus bacteria in the intestinal tract microflora of carps. Biologija, 2: 13- 17.
Kandler, O, and Weiss, N. 1986. In: P.H.A. Sneath, N.S. Mair, M.E. Sharpe and J.G. Holt (Ed.), Bergey’s Manual of Systematic Bacteriology. Vol 2, Bathimore: Williams and Wilkins (1994), 1209-1234.
Karimi Torshizi, M. A., Rahimi, S., Mojgani, N., Esmaeilkhanian, S. and Grimes, J.L. 2008. Screening of indigenous strains of lactic acid bacteria for development of a probiotic for poultry. Asian– Australasian Journal of Animal Sciences, 21: 1495- 1500.1151. doi:10.1111/j.1365-2672.2005.02546.x
Trousselier, M. and Legendre, P. 1981. A functional evenness index for microbial ecology. Microbial Ecology, 7: 283-296. doi: 10.1007/BF02341423
Xiraphi, N., Georgalaki, M., Rantsiou, K., Cocolin, L., Tsakalidou, E. and Drosinos, E.H. 2008. Purification and characterization of a bacteriocin produced by Leuconostoc mesenteroides E131. Meat Science, 80: 194-203. doi: 10.1016/j.meatsci.2007.11.020
Yu, J., Gao, W., Qing, M., Sun, Z., Wang, W., Liu, W., Pan, L., Sun, T., Wang, H., Bai, N. and Zhang, H. 2012. Identification and characterization of lactic acid bacteria isolated from traditional pickles in Sichuan, China. Journal of General and Applied Microbiology, 58:163-172. doi. 10.2323/jgam.58.163