Diseases in aquatic organisms, including fish, are a major concern in aquaculture production. In this present investigation, we have evaluated the beneficial effects of dietary Bacillus amyloliquefaciens CCF7 (GenBank Acc. No. KP256501) supplementation in rohu (Labeo rohita) challenged by a pathogenic strain of Aeromonas hydrophila MTCC 1739. Four experimental diets were formulated: control diet (no probiotics) and three experimental diets (different concentrations of probiotic candidate B. amyloliquefaciens CCF7 at 10 (T1), 10 (T2), 10 (T3) CFU/g). Further, we have divided the feeding trial into pre-challenge (70 days) and post-challenge (28 days) periods and various immune parameters (serum protein, globulin, albumin, lysozyme, and IgM), and stress parameters (malondialdehyde, catalase, and superoxide dismutase) were examined during both the periods. Throughout the entire experiment, control group was fed with probiotic free basal diet, while the treatment groups received probiotic supplemented diets (PSD). After challenge test, serum aspartate transaminase (AST), serum alanine transaminase (ALT) activity, and liver malondialdehyde level have increased significantly in control groups; however, level of these parameters were considerably lower in fish fed with PSD. In contrast, liver catalase and superoxide dismutase activities and serum globulin concentration was significantly higher in the group fed with T3 diet followed by T2. Furthermore, an elevated level of serum IgM and higher activity of serum lysozyme was also recorded in PSD fed groups, especially for T3 group which confirmed the probiotic efficiency of the bacterium B. amyloliquefaciens CCF7. We strongly believe that B. amyloliquefaciens CCF7 will be a good probiotic candidate in aquaculture industries.

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In this study, a total of 121 bacterial strains were isolated from the gastrointestinal tract of four teleostean species, namely striped snakehead (Channa striatus), striped dwarf catfish (Mystus vittatus), orangefin labeo (Labeo calbasu) and mrigal carp (Cirrhinus mrigala), among which 8 isolates showed promising antibacterial activity against four potential fish pathogens, Aeromonas hydrophila, Aeromonas salmonicida, Aeromonas sobria and Pseudomonas fluorescens and were non-hemolytic. The isolates were further screened in response to fish bile tolerance and extracellular digestive enzyme activity. Two bacterial strains MVF1 and MVH7 showed highest tolerance and extracellular enzymes activities, and selected for further studies. Antagonistic activity of these two isolates was further confirmed by in vitro growth inhibition assay against four selected fish pathogens in liquid medium. Finally, these two bacterial strains MVF1 and MVH7 were selected as potential probiotic candidates and thus identification by partial 16S rRNA gene sequence analysis. The bacterial isolates MVF1 and MVH7 were identified as two strains of Bacillus sp.

A b s t r a c tScanning electron microscopy (SEM) was used to define the location of epithelium-associated bacteria in the gastrointestinal (GI) tract of two Indian air-breathing fish, the climbing perch (Anabas testudineus) and walking catfish (Clarias batrachus). The SEM examination revealed substantial numbers of rod shaped bacterial cells associated with the microvillus brush borders of enterocytes in proximal (PI) and distal regions (DI) of the GI tract of both the fish species. Ten (two each from the PI and DI of climbing perch and three each from the PI and DI of walking catfish) isolated bacterial strains were evaluated for extracellular protease, amylase and cellulase production quantitatively. All the bacterial strains exhibited high cellulolytic activity compared to amylolytic and proteolytic activites. Only two strains, CBH6 and CBH7, isolated from the DI of walking catfish exhibited high proteolytic activity. Maximum cellulase activity was exhibited by the strain, CBF2, isolated from the PI of climbing perch. Six most promising enzyme-producing adherent bacterial strains were identified by 16S rDNA gene sequence analysis. The strain ATH1 (isolated from climbing perch) showed high similarity to Bacillus amyloliquefaciens whereas, the remaining five strains (isolated from walking catfish) were most closely related to Bacillus licheniformis. K e y w o r d s: 16S rDNA, air-breathing fish, enzyme production identification, GI tract bacteria identification, SEM