The fish disease epidemiology is urgent to be investigated for the surveillance and prevention. The diseased fish showed splenomegaly with diffusion of white nodules and microscopical granulomatous formation. It is important to develop a method of pathogens isolated from clinical samples with serial dilution method and disc diffusion method. Representative colonies were selected from diseased cobia on BHIA plate and were inoculated onto MacConkey agar, TCBS agar, and blood agar. The cage-culture of the different bacterial groups detected in the survey of bacteria isolated from THOD, HDSB, EMD with serial dilution method. 119 from 128 isolated strains were Gram¡¦s negative (93%), including pathogenic Vibrio spp. 57% (73/128) in THOD. 54 from 90 isolated strains were Gram¡¦s negative (60%), including pathogenic Vibrio spp. 12.2% (11/90) in HDSB. 61 from 104 isolated strains were Gram¡¦s negative (59%), including pathogenic Vibrio spp. 70.2% (77/90) in EMD. In different times diseased grouper, 104 from 139 isolated strains were Gram¡¦s negative (75%), including pathogenic Vibrio spp. 88% (123/139), in 2003. While 24 from 44 isolated strains were Gram¡¦s negative (55%), including pathogenic Vibrio spp. 73% (32/44), in 2004. 66 from 75 isolated strains were Gram¡¦s negative (88%), including pathogenic Vibrio spp. 97% (73/75) by disc dilution method in EMD. 9 from 31 isolated strains were Gram¡¦s negative (30%), including pathogenic Vibrio spp. 26% (8/31), by disc dilution method of grouper in PCG. A DGGE (denaturing gradient gel electrophoresis) technique can identify six groups of bacteria from cobia, and J6, R13, T29 have similarity 100%. Quantity One Version 4.5 (Bio-Rad) can identify six groups of bacteria from diffusion methods that F group diluted the bacterial strain from serial dilution method. B group and E group diluted the bacterial strain from disc diffusion method. Higher resistance rates of the different bacterial strains isolated from cobia and were £]-lactam and susceptible were observed in quinolones.

Antibiotic resistance is a global health threat and consequently, there is a need to understand the mechanisms driving its emergence. Here, we hypothesize that genes and mutations under positive selection may contribute to antibiotic resistance. We explored wastewater E. coli, whose genomes are highly diverse. We subjected 92 genomes to a statistical analysis for positively selected genes. We obtained 75 genes under positive selection and explored their potential for antibiotic resistance. We found that eight genes have functions relating to antibiotic resistance, such as biofilm formation, membrane permeability, and bacterial persistence. Finally, we correlated the presence/absence of non-synonymous mutations in positively selected sites of the genes with a function in resistance against 20 most prescribed antibiotics. We identified mutations associated with antibiotic resistance in two genes: the porin ompC and the bacterial persistence gene hipA. These mutations are located at the surface of the proteins and may hence have a direct effect on structure and function. For hipA, we hypothesize that the mutations influence its interaction with hipB and that they enhance the capacity for dormancy as a strategy to evade antibiotics. Overall, genomic data and positive selection analyses uncover novel insights into mechanisms driving antibiotic resistance.