Impact of polymorphisms in blaZ, blaR1 and blaI genes and their relationship with ß-lactam resistance in S. aureus strains isolated from bovine mastitis.

There is not a consensus between the presence of the genotypic resistance marker gene and the phenotypic resistance to ß-lactams in Staphylococcus aureus, which means, positive S. aureus blaZ isolates demonstrating sensitivity to ß-lactams. The present study aimed to characterize the blaZ, blaR1 and blaI genes, identify and evaluate single nucleotide polymorphisms (SNPs) and their relationship with ß-lactam resistance in samples of Staphylococcus aureus obtained from cases of bovine mastitis. Five isolates (two resistant and three sensitive to oxacillin) of Staphylococcus aureus with detected production of beta-lactamase, previously evaluated as containing the blaZ gene and negative for the mecA and mecC genes, had the bla operon completely sequenced. Impacts on the protein sequence due to the detected polymorphisms were evaluated by modeling the proteins encoded by the blaZ, blaR1 and blaI genes using a three-dimensional model structure obtained from the Protein Data Bank (PDB) database. Fifteen SNPs were detected in the blaZ gene, 30 in the blaR1 gene and three in the blaI gene. These SNPs caused alterations in amino acid sites. Deleterious mutations were detected in the blaZ gene (E146G, P218S, Y221C) and the blaR1 gene (K481E). Molecular docking analysis revealed that polymorphisms in the blaZ gene may explain the phenotypic sensitivity in isolates that contain the resistance marker gene. Although sensitive and resistant isolates encode beta-lactamase, these proteins are functionally altered due to a change in the binding site with the antibiotic.

Resistance and virulence in Staphylococcus aureus by whole-genome sequencing: a comparative approach in blaZ-positive isolates.

Mastitis caused by Staphylococcus aureus is a worldwide problem in dairy farms, in part because of the pathogenicity of the bacteria, biofilm formation, and mechanisms of antimicrobial resistance that make the disease difficult to diagnose and treat, which is typically done with the use of beta-lactam antibiotics. The aim of the present study was to determine the virulence and resistance factors of S. aureus isolates from subclinical mastitis, blaZ + /mecA - /mecC - , resistant and sensitive to oxacillin. All isolates were classified as CC97 by MLST analysis, a clonal complex well adapted to the mammary gland and although STAU23 and STAU73 were resistant to oxacillin while STAU32 and STAU78 were sensitive, the genomic analysis identified only the blaZ operon corresponding to resistance to beta-lactams. However, the presence of the sdrC gene was revealed exclusively in resistant isolates, an important adhesin in the colonization process that potentiates pathogenicity in S. aureus. In addition, resistance islands (REIs) were identified in these isolates, suggesting more conserved REIs. In the analysis of SNPs throughout the genome, mutations were found in the trmB and smpB genes of the resistant isolates and in the murD and rimM genes of the sensitive isolates. This study highlights the potential benefit of genome-wide characterization tools to identify molecular mechanisms of S. aureus in bovine mastitis.

Assessment of clinical characteristics and viral load in individuals infected by Delta and Omicron variants of SARS-CoV-2.

In late 2021, a new variant of SARS-CoV-2 called Omicron emerged, replacing Delta worldwide. Although it has been associated with a lower risk of hospitalization and severe forms of COVID-19, there is little evidence of its relationship with specific symptoms and viral load. The aim of this study was to verify the relationship between Delta and Omicron variants of concern, viral load, and the occurrence of symptoms in individuals with COVID-19. Nasopharyngeal swab samples were collected and sequenced from patients with COVID-19 from the Northeast Region of Brazil between August 2021 and March 2022. The results showed a gradual replacement of the Delta variant by the Omicron variant during the study period. A total of 316 samples (157 Delta and 159 Omicron) were included. There was a higher prevalence of symptoms in Delta-infected individuals, such as coryza, olfactory and taste disturbances, headache, and myalgia. There was no association between viral load and the variants analyzed. The results reported here contribute to the understanding of the symptoms associated with the Delta and Omicron variants in individuals affected by COVID-19.