Salmonellae display intricate evolutionary patterns comprising over 2500 serovars having diverse pathogenic profiles. The acquisition and/or exchange of various virulence factors influences the evolutionary framework. To gain insights into evolution of Salmonella in association with the CRISPR-Cas genes we performed phylogenetic surveillance across strains of 22 Salmonella serovars. The strains differed in their CRISPR1-leader and cas operon features assorting into two main clades, CRISPR1-STY/cas-STY and CRISPR1-STM/cas-STM, comprising majorly typhoidal and non-typhoidal Salmonella serovars respectively. Serovars of these two clades displayed better relatedness, concerning CRISPR1-leader and cas operon, across genera than between themselves. This signifies the acquisition of CRISPR1/Cas region could be through a horizontal gene transfer event owing to the presence of mobile genetic elements flanking CRISPR1 array. Comparison of CRISPR and cas phenograms with that of multilocus sequence typing (MLST) suggests differential evolution of CRISPR/Cas system. As opposed to broad-host-range, the host-specific serovars harbor fewer spacers. Mapping of protospacer sources suggested a partial correlation of spacer content with habitat diversity of the serovars. Some serovars like serovar Enteritidis and Typhimurium that inhabit similar environment/infect similar hosts hardly shared their protospacer sources.
Microbial exopolysaccharides (EPS) provide a broad range of applications. Thus, there is an increasing interest in the production, characterization, and use of EPS derived from various microorganisms. Extremophile polysaccharides have unique properties and applications due to its unique structures. The importance of exopolysaccharides synthesized by a new bacterial strain, Alkalibacillus sp. w3, was highlighted in this study. Alkalibacillus sp. w3, a haloalkalitolerant firmicute that was recovered from a salt lake, was optimized for EPS production, and its biological activities were studied. Exopolysaccharide synthesis was observed in Horikoshi I broth medium. The optimal culture conditions for achieving the highest exopolysaccharide production were a 7-day incubation period, pH 10, and 250 g/L of NaCl. The most effective carbon and nitrogen sources for EPS production were glucose and a combination of yeast extract and peptone. Additionally, Plackett-statistical Burman’s design showed that all factors tested had a favorable impact, with glucose having the greatest significance on the production of EPS. The model’s best predictions for culture conditions resulted in a two-fold improvement in EPS production compared to the original yield before optimization. The recovered EPS contained 65.13% carbohydrates, 30.89% proteins, and 3.98% lipids. Moreover, EPS produced by Alkalibacillus sp. w3 demonstrated anticancer activity against hepatocellular carcinoma (HepG2) and human colon carcinoma (HCT-116) cell lines, with IC50 values as low as 11.8 and 15.5 µg/mL, respectively, besides antibacterial activity against various Gram-positive, Gram-negative bacteria, and yeast. Based on these results, EPS made by Alkalibacillus sp. w3 has many useful properties, which make it suitable for use in the medical field.
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