Complete genome and plasmid sequence of a novel Bacillus sp. BD59S, a parasporal protein synthesizing bacterium.

Parasporal crystalline inclusion proteins of some Bacillus spp. are of paramount importance due to their insecticidal, nematocidal, and cancer cell killing capabilities. Here, we present a brief report of the complete genome sequence of Bacillus sp. BD59S, a bacterium that produced HeLa cell-killing parasporal crystalline inclusion proteins. From genome sequencing and assembly, we found that the bacterium has one circular chromosome and two large plasmids, pBTBD59S1 and pBTBD59S2. The size of the chromosome is 5283,933 bp with a 35.4% GC content, consisting of 5938 genes and 5550 protein-coding sequences (CDSs), 25 complete rRNAs (5S, 16S, 23S), 98 tRNAs, 5 ncRNAs, 260 pseudo-genes, and 356 subsystems. Complete plasmid sequence of pBTBD59S1 comprises a total size of 162,149 bp with 33.4% GC content, 192 CDSs, and 13 subsystems. The other plasmid pBTBD59S2, is 199,209 bp long with 32.9% GC content, 179 CDSs, and 11 subsystems. Analyses by NCBI microbial genome BLAST, phylogenetic genome tree, and BLAST ring image generator (BRIG) revealed that BD59S belongs to Bacillus cereus group, and is more close to B. thuringiensis. Further, the strain possesses 57.04 kDa and 54.42 kDa Cry protein-coding genes, which show significant similarities with cancer cell-killing parasporin proteins of B. thuringiensis strains.

Partial purification and characterization of serine protease produced through fermentation of organic municipal solid wastes by Serratia marcescens A3 and Pseudomonas putida A2.

Proteolytic bacteria isolated from municipal solid wastes (MSW) were identified as Serratia marcescens A3 and Pseudomonas putida A2 based on 16S rDNA sequencing. Protease produced through fermentation of organic MSW by these bacteria under some optimized physicochemical parameters was partially purified and characterized. The estimated molecular mass of the partially purified protease from S. marcescens and P. putida was approximately 25 and 38 kDa, respectively. Protease from both sources showed low Km 0.3 and 0.5 mg ml-1 and high Vmax 333 and 500 µmole min-1 at 40 °C, and thermodynamics analysis suggested formation of ordered enzyme-substrate (E-S) complexes. The activation energy (Ea) and temperature quotient (Q10) of protease from S. marcescens and P. putida were 16.2 and 19.9 kJ/mol, and 1.4 and 1.3 at temperature range from 20 to 40 °C, respectively. Protease of the both bacterial isolates was serine and cysteine type. The protease retained approximately 97% of activity in the presence of sodium dodecyl sulphate. It was observed that the purified protease of S. marcescens could remove blood stains from white cotton cloth and degrade chicken flesh remarkably. Our study revealed that organic MSW can be used as raw materials for bacterial protease production and the protease produced by S. marcescens A3 might be potential for applications.

Vibrio cholerae O1 with Reduced Susceptibility to Ciprofloxacin and Azithromycin Isolated from a Rural Coastal Area of Bangladesh.

Cholera outbreaks occur each year in the remote coastal areas of Bangladesh and epidemiological surveillance and routine monitoring of cholera in these areas is challenging. In this study, a total of 97 Vibrio cholerae O1 isolates from Mathbaria, Bangladesh, collected during 2010 and 2014 were analyzed for phenotypic and genotypic traits, including antimicrobial susceptibility. Of the 97 isolates, 95 possessed CTX-phage mediated genes, ctxA, ace, and zot, and two lacked the cholera toxin gene, ctxA. Also both CTX+ and CTX-V. cholerae O1 isolated in this study carried rtxC, tcpAET, and hlyA. The classical cholera toxin gene, ctxB1, was detected in 87 isolates, while eight had ctxB7. Of 95 CTX+V. cholerae O1, 90 contained rstRET and 5 had rstRCL. All isolates, except two, contained SXT related integrase intSXT. Resistance to penicillin, streptomycin, nalidixic acid, sulfamethoxazole-trimethoprim, erythromycin, and tetracycline varied between the years of study period. Most importantly, 93% of the V. cholerae O1 were multidrug resistant. Six different resistance profiles were observed, with resistance to streptomycin, nalidixic acid, tetracycline, and sulfamethoxazole-trimethoprim predominant every year. Ciprofloxacin and azithromycin MIC were 0.003-0.75 and 0.19-2.00 µg/ml, respectively, indicating reduced susceptibility to these antibiotics. Sixteen of the V. cholerae O1 isolates showed higher MIC for azithromycin (≥0.5 µg/ml) and were further examined for 10 macrolide resistance genes, erm(A), erm(B), erm(C), ere(A), ere(B), mph(A), mph(B), mph(D), mef(A), and msr(A) with none testing positive for the macrolide resistance genes.