Genomic comparisons and phylogenetic analysis of mastitis‑related staphylococci with a focus on adhesion, bioflm, and related regulatory genes.

Mastitis is a common and costly disease on dairy farms, commonly caused by Staphylococcus spp. though the various species are associated with different clinical outcomes. In the current study, we performed genomic analyses to determine the prevalence of adhesion, biofilm, and related regulatory genes in 478 staphylococcal species isolated from clinical and subclinical mastitis cases deposited in public databases. The most prevalent adhesin genes (ebpS, atl, pls, sasH and sasF) were found in both clinical and subclinical isolates. However, the ebpS gene was absent in subclinical isolates of Staphylococcus arlettae, S. succinus, S. sciuri, S. equorun, S. galinarum, and S. saprophyticus. In contrast, the coa, eap, emp, efb, and vWbp genes were present more frequently in clinical (vs. subclincal) mastitis isolates and were highly correlated with the presence of the biofim operon (icaABCD) and its transcriptional regulator, icaR. Co-phylogenetic analyses suggested that many of these adhesins, biofilm, and associated regulatory genes could have been horizontally disseminated between clinical and subclinical isolates. Our results further suggest that several adhesins, biofilm, and related regulatory genes, which have been overlooked in previous studies, may be of use for virulence profiling of mastitis-related Staphylococcus strains or as potential targets for vaccine development

Association between coa gene and enterotoxin gene in S. aureus from dairy cattle in Brazil.

ABSTRACT - Staphylococcus aureus is an important agent in bovine mastitis, and some specific virulence factors may be implicated in this disease. Therefore, this study aimed to investigate the importance of the presence of coagulase, superantigens, genotypic and phenotypic resistance, and pulsotypes in 65 S. aureus isolates from bovine clinical and subclinical mastitis in the Southeast of Brazil. A high correlation was observed between the genes coa and see, as well as between the sei and the see and seh. High resistance rates were observed for penicillin (95.4%), tetracycline (89.2%), cefoxitin (86.1%), oxacillin (84.6%), erythromycin (84.6%), clindamycin (84.6%), chloramphenicol (81.5%), ceftriaxone (80.0%), and ampicillin (80.0%). Analysis of antimicrobial resistance profiles showed that 89.2% of isolates were multi-drug-resistant. No mecA-positive S. aureus isolates were detected. It was observed that seven isolates were resistant to all the β-lactam tested while being susceptible to cefoxitin, which could be indicative of borderline methicillin resistance in S. aureus. High genetic diversity with no specific virulence profile being predominant was observed. Thus, this study observed a high correlation between the coa and enterotoxins genes, and demonstrates that there is no predominant pulsotype causing intramammary infection and that there is a high rate of antibiotic resistance in S. aureus isolates from dairy farms in the southeast regions of Brazil

Allosteric modulation of pyrophosphatase activity of rat osseous plate alkaline phosphatase by magnesium ions

Pyrophosphatase activity of rat osseous plate alkaline phosphatase was studied at different concentrations of calcium and magnesium ions. with the aim of characterizing the modulation of enzyme activity by these metals. In the absence of metal ions, the enzyme hydrolysed pyrophosphate following Michaelian kinetics with a specific activity of 36.7 U/mg and K-0.5 = 88 mu M. In the presence of low concentrations (0.1 mM) of magnesium (or calcium) ions, the enzyme also exhibited Michaclian kinetics for the hydrolysis of pyrophosphate, but a significant increase in specific activity (123 U/mg) was observed. K-m values remained almost unchanged. Quite different behavior occurred in the presence of 2 mM magnesium (or calcium) ions. In addition to low-affinity sites (K-0.5 = 40 and 90 mu M, for magnesium and calcium, respectively), high-affinity sites were also observed with K-0.5 values 100-fold lower. The high-affinity sites observed in the presence of calcium ions represented about 10% of those observed for magnesium ions. This was correlated with the fact that only magnesium ions triggered conformational changes yielding a fully active enzyme. These results suggested that the enzyme could hydrolyse pyrophosphate, even at physiological concentrations (4 mu M), since magnesium concentrations are high enough to trigger conformational changes increasing the enzyme activity. A model, suggesting the involvement of magnesium ions in the hydrolysis of pyrophosphate by rat osseous plate alkaline phosphatase is proposed. (C) 1998 Published by Elsevier B.V. Ltd. All rights reserved