Microbes evolve rapidly by modifying their genomes through mutations or through the horizontal acquisition of mobile genetic elements (MGEs) linked with fitness traits such as antimicrobial resistance (AMR), virulence, and metabolic functions. We conducted a multicentric study in India and collected different clinical samples for decoding the genome sequences of bacterial pathogens associated with sepsis, urinary tract infections, and respiratory infections to understand the functional potency associated with AMR and its dynamics. Genomic analysis identified several acquired AMR genes (ARGs) that have a pathogen-specific signature. We observed that bla CTX-M-15 , bla CMY-42 , bla NDM-5 , and aadA (2) were prevalent in Escherichia coli , and bla TEM-1B , bla OXA-232 , bla NDM-1 , rmtB , and rmtC were dominant in Klebsiella pneumoniae . In contrast, Pseudomonas aeruginosa and Acinetobacter baumannii harbored bla VEB , bla VIM-2 , aph( 3’), strA/B , bla OXA-23 , aph (3′) variants, and amrA , respectively. Regardless of the type of ARG, the MGEs linked with ARGs were also pathogen-specific. The sequence type of these pathogens was identified as high-risk international clones, with only a few lineages being predominant and region-specific. Whole-cell proteome analysis of extensively drug-resistant K. pneumoniae , A. baumannii, E. coli, and P. aeruginosa strains revealed differential abundances of resistance-associated proteins in the presence and absence of different classes of antibiotics. The pathogen-specific resistance signatures and differential abundance of AMR-associated proteins identified in this study should add value to AMR diagnostics and the choice of appropriate drug combinations for successful antimicrobial therapy.
Nanoparticles show the multidisciplinary versatile utility and are gaining the prime place in various fields, such as medicine, electronics, pharmaceuticals, electrical designing, cosmetics, food industries, and agriculture, due to their small size and large surface to volume ratio. Biogenic or green synthesis methods are environmentally friendly, economically feasible, rapid, free of organic solvents, and reliable over conventional methods. Plant extracts are of incredible potential in the biosynthesis of metal nanoparticles owing to their bountiful availability, stabilizing, and reducing ability. In the present study, the aqueous leaf extract of Buchanania lanzan Spreng was mixed with 0.5 mM silver nitrate and incubated at 70°C for 1 h and synthesized a good quantity of AgNPs. The synthesized AgNPs were characterized using UV-visible spectroscopy, X-ray diffractometry (XRD), dynamic light scattering (DLS), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). The maximum absorption of UV-visible spectra was obtained in the range of 420–430 nm. Furthermore, SEM and TEM results inferred that the size of the particles were 23–62 nm, spherical, crystalline, uniformly distributed, and negatively charged with the zeta potential of −27.6 mV. In addition, the antifungal activities of the AgNPs were evaluated against two phytopathogenic fungi Rhizoctonia solani and Fusarium oxysporum f. sp. lycopersici in vitro using poison food techniques on PDA media. The maximum rate of mycelia inhibition was found in 150 ppm concentration of AgNPs against both phytopathogenic fungi.
Vibrio cholerae O1 and O139 isolates deploy cholera toxin (CT) and toxin-coregulated pilus (TCP) to cause the diarrhoeal disease cholera. The ctxAB and tcpA genes encoding CT and TCP are part of two acquired genetic elements, the CTX phage and Vibrio pathogenicity island-1 (VPI-1), respectively. ToxR and ToxT proteins are the key regulators of virulence genes of V. cholerae O1 and O139. V. cholerae isolates belonging to serogroups other than O1/O139, called non-O1/non-O139, are usually devoid of virulence-related elements and are non-pathogenic. Here, we have analysed the available whole genome sequence of an environmental toxigenic V. cholerae non-O1/non-O139 strain, VCE232, carrying the CTX phage and VPI-1. Extensive bioinformatics and phylogenetic analyses indicated high similarity of the VCE232 genome sequence with the genome of V. cholerae O1 strains, including organization of the VPI-1 locus, ctxAB, tcpA and toxT genes, and promoters. We established that the VCE232 strain produces an optimal amount of CT at 30 °C under AKI conditions. To investigate the role of ToxT and ToxR in the regulation of virulence factors, we constructed ΔtoxT, ΔtoxR and ΔtoxTΔtoxR deletion mutants of VCE232. Extensive genetic analyses of these mutants indicated that the toxT and toxR genes of VCE232 are crucial for CT and TCP production. However, unlike O1 isolates, the presence of either toxT or toxR gene is sufficient for optimal CT production in VCE232. In addition, the VCE232 ΔtoxR mutant showed differential regulation of the major outer membrane proteins, OmpT and OmpU. This is the first attempt to explore the regulation of expression of major virulence genes and regulators in an environmental toxigenic V. cholerae non-O1/non-O139 strain.
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