Effect of pH and salinity on pathogenicity of Flavobacterium columnare and Myxobacterium sp. in Indian cat fish, Clarias batrachus (Linn) and Heteropneustes fossilis (Bloch).

Flavobacterium columnare (FC) and Myxobacterium sp. recorded persistently associated in fish hatchery and culture system of Himalayan and Sub - Himalayan regions were found to be pathogenic. The pH and salinity played a significant role on the pathogenicity of these potent pathogens in case of Clarias batrachus and Heteropneustes fossilis. LD50 value of FC was 104.5 CFU in both the fishes and those of Myxobacterium sp it was 106 CFU ml-1 fish-1. Fish challenged with F. columnare and Myxobacterium sp. (@ 0.2 ml fish-1) individually consisting 105-6cfu ml-1 exhibited explicit symptoms of columnaris disease and marked with ulceration and saddle back lesion on the dorsal side of body. Maximum reisolation of inoculated bacteria was recorded at pH 7.0 and 7.5 and at 0.0-0.5 (F. columnare) and 0.0-1.0% (Myxobacterium sp.) salinity. Foregoing results elucidated that F. columnare was more sensitive to salinity in comparison to Myxobacterium sp. and their pathogenicity significantly (p<0.05) depends on the salinity and pH that might be one of the physical factors to control their proliferation.

Lytic bacteriophages specific to Flavobacterium columnare rescue catfish, Clarias batrachus (Linn.) from columnaris disease.

This investigation was aimed to find out appropriate strategy against antibiotic resistant bacterial fish pathogen, F. columnare. This pathogen was found persistently associated with fishes causing columnaris disease and ensuing mass mortality in hatchery and culture system of Sub - Himalayan region. Nine lytic F. columnare phages (FCP1 – FCP9) specific to its fifteen isolates were isolated from the water and bottom sediments of various geo-climatic regions of North India. The F. columnare phage FCP1 (made of hexagonal head and non contractile long tail belonging to family Podovariedae, a member of DNA virus) exhibited broader host range to lyse 9 out of 15 isolates of F. columnare. Therapeutic ability of FCP1 phage was assessed in C. batrachus inoculated intramuscularly (im) with virulent bacterial isolate FC8 and post inoculated (PI) with FCP1phage (@ 108 : 106 :: cfu : pfu) through intramuscular (im), immersion (bath) and oral (phage impregnated feed) treatment. Significant (p<0.001) reduction (less than 10-3 cfu ml-1) in host bacterium in the sera, gill, liver and kidney of challenged fishes was noted after 6 hr of phage treatment. Quantum of phage played a significant role in bringing down bacterial population as in the sera of dose 1 (@ 4.55 X 106 pfu ml-1) and dose 2 (@ 9.15 X 106 pfu ml-1) treated fishes mean log10 cfu value reduced by 3 logs (58.39%) and 5 logs (73.77%) at 96 hr, respectively. Phage treatment led to disappearance of gross symptoms, negative bacteriological test, detectable phage and 100% survival in experimentally infected C. batrachus. Result of this study provides evidence of profound lytic impact of FCP1 phage and represents its interesting therapeutic importance against antibiotic resistant F. columnare.