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Culturable autochthonous gut bacteria in rohu, Labeo rohita. In vitro growth inhibition against pathogenic Aeromonas spp., stability in gut, bio-safety and identification by 16S rRNA gene sequencing

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Abstract

Autochthonous endosymbiotic gut bacteria antagonistic against pathogenic Aeromonas spp. have been evaluated in rohu, Labeo rohita for characterization of putative probiotics. Four promising pathogen inhibitory bacteria (23 strains out of 225 isolates showed antagonism) were selected by double layer assay, following which inhibition pattern was examined through in vitro growth curve and statistical analyses. Cell free supernatant (CFS) of the selected gut bacteria significantly inhibited the growth of pathogenic aeromonads. While, CFS of strain LR3FG26 was the most efficient among them. Selected strains were γ haemolytic and susceptible to most of the common antibiotics that demonstrated their likely non-pathogenic and eco-friendly nature. Additionally, selected bacteria produced different exo-enzymes (digestive and antinutritional factors degrading), could grow better in skin mucus than intestinal mucus (exceptionally, LR3FG26 grew better in skin mucus) and tolerated diluted bile juice (2–20%). 16S rRNA partial gene sequence analyses and Blast search in NCBI GenBank unveiled that the strains LR1FG1, LR2HG13, LR3FG26 and LR3HG4 were similar with the type strains of Bacillus methylotrophicus (NR116240), Bacillus amyloliquefaciens (NR117946), Pseudomonas fluorescens (NR113647) and Bacillus licheniformis (NR118996), respectively. Application of these symbiotic pathogen-inhibitory bacteria in pathogen challenge studies is required to appraise their probiotic effects in vivo.

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Acknowledgements

The first author is obliged to the DST-INSPIRE programme for the research fellowship. The authors are grateful to Head of Department of Zoology, The University of Burdwan, West Bengal, India; The University Grants Commission (UGC-SAP-DRS programme), New Delhi, India and The Department of Science and Technology (FIST programme), New Delhi, India for research support. The authors are grateful to Dr. G. Aditya for rendering help in statistical analyses of data.

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Authors and Affiliations

  1. Aquaculture Laboratory, Department of Zoology, The University of Burdwan, Golapbag, Burdwan, West Bengal, 713104, India

    Anjan Mukherjee, Dipanjan Dutta, Sudeshna Banerjee & Koushik Ghosh

  2. Norwegian College of Fishery Science, Faculty of Biosciences, Fisheries and Economics, UiT The Arctic University of Norway, -9037, Tromsø, NO, Norway

    Einar Ringø

  3. Department of Arctic and Marine Biology, Faculty of Bioscience, Fisheries and Economics, UiT The Arctic University of Norway, -9037, Breivika, NO, Norway

    Eva Marie Breines & Ellinor Hareide

  4. Parasitology Laboratory, Department of Zoology, The University of Burdwan, Burdwan, West Bengal, 713104, India

    Goutam Chandra

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  1. Anjan Mukherjee
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  2. Dipanjan Dutta
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  3. Sudeshna Banerjee
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Mukherjee, A., Dutta, D., Banerjee, S. et al. Culturable autochthonous gut bacteria in rohu, Labeo rohita. In vitro growth inhibition against pathogenic Aeromonas spp., stability in gut, bio-safety and identification by 16S rRNA gene sequencing. Symbiosis 73, 165–177 (2017). https://doi.org/10.1007/s13199-017-0474-7

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