Introduction: Methicillin-resistant Staphylococcus aureus (MRSA) presents a profound hazard to public health. MRSA colonizing skin, mucous membranes, and the anterior nares without clinical symptoms is termed "colonizing MRSA". Upon manifestation of clinical symptoms, it is termed "infectious MRSA". Here, we characterize and differentiate colonizing and infectious MRSA, and analyze the phenotypic-genotypic and antibiotic susceptibility correlations. Methodology: Clinical MRSA isolates were recovered from intensive care units (ICUs) of two major Egyptian hospitals and their biofilm formation ability was tested. Antibiograms against 16 antibiotics were determined, in addition to the minimum inhibitory concentrations (MICs) of vancomycin and linezolid. The entire collection was typed by enterobacterial repetitive intergenic consensus (ERIC)-PCR, as well as multilocus sequence typing (MLST). Representative resistance and virulence genes were detected by PCR amplification. Results: Forty-nine isolates were confirmed as MRSA, of which 30 isolates were infectious and 19 were colonizing. Versatile resistance patterns were observed in both groups of isolates. We report a higher tendency for biofilm-formation and borderline minimum inhibitory concentrations among infectious isolates. A Positive antibiotic correlation was observed between susceptibility to protein synthesis inhibitors and cell wall inhibitors. Positive correlations were observed between isolation site and rifampicin resistance: nasal samples were enriched in rifampicin-resistant isolates, while urine and blood samples were enriched in susceptible ones. Furthermore, biofilm formation ability was slightly associated with amikacin resistance, and an association between teicoplanin resistance and the presence of the Panton-Valentine leukocidin gene was the only significant phenotype-genotype correlation observed. Finally, ERIC typing and MLST had congruent results. Conclusion: Linezolid and vancomycin are still the most convenient choice for MRSA treatment. ERIC PCR and MLST show promising typing combination that could be easily used periodically for tracking the genotypic changes of MRSA, especially within the healthcare facilities. Several correlations were established between groups of antibiotics and the genotypes/phenotypes of the selected isolates.
Staphylococcus aureus is a Gram-positive pathogen that is capable of infecting almost every organ in the human body. Alarmingly, the rapid emergence of methicillin-resistant S. aureus strains (MRSA) jeopardizes the available treatment options. Herein, we propose sustainable, low-cost production of recombinant lysostaphin (rLST), which is a native bacteriocin destroying the staphylococcal cell wall through its endopeptidase activity. We combined the use of E. coli BL21(DE3)/pET15b, factorial design, and simple Ni-NTA affinity chromatography to optimize rLST production. The enzyme yield was up to 50 mg/L culture, surpassing reported systems. Our rLST demonstrated superlative biofilm combating ability by inhibiting staphylococcal biofilms formation and detachment of already formed biofilms, compared to vancomycin and linezolid. Furthermore, we aimed at developing a novel rLST topical formula targeting staphylococcal skin infections. The phase inversion composition (PIC) method fulfilled this aim with its simple preparatory steps and affordable components. LST nano-emulgel (LNEG) was able to extend active LST release up to 8 h and cure skin infections in a murine skin model. We are introducing a rapid, convenient rLST production platform with an outcome of pure, active rLST incorporated into an effective LNEG formula with scaling-up potential to satisfy the needs of both research and therapeutic purposes.
Cholera is a devastating diarrheal disease that accounts for more than 10% of children's lives worldwide, but its treatment is hampered by a rise in antibiotic resistance. One promising alternative to antibiotic therapy is the use of bacteriophages to treat antibiotic-resistant cholera infections, and control Vibrio cholera in clinical cases and in the environment, respectively. Here, we report four novel, closely related environmental myoviruses, VP4, VP6, VP18, and VP24, which we isolated from two environmental toxigenic Vibrio cholerae strains from river Kuja and Usenge beach in Kenya. High-throughput sequencing followed by bioinformatics analysis indicated that the genomes of the four bacteriophages have closely related sequences, with sizes of 148,180 bp, 148,181 bp, 148,179 bp, and 148,179 bp, and a G + C content of 36.4%. The four genomes carry the phoH gene, which is overrepresented in marine cyanophages. The isolated phages displayed a lytic activity against 15 environmental, as well as one clinical, Vibrio cholerae strains. Thus, these novel lytic vibriophages represent potential biocontrol candidates for water decontamination against pathogenic Vibrio cholerae and ought to be considered for future studies of phage therapy.
Twenty-three compounds of novel phenylthiazole derivatives were synthesized utilizing the Suzuki coupling reaction. Antibacterial activity in an in vivo model against MRSA USA300 was evaluated, as it reduced the burden of MRSA USA300 in skin-infected mice.
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