Staphylococcus aureus strains were isolated from mastitic milk of cows with infected mammary glands. The animals were living in 12 different farms near Lublin, in Central-Eastern Poland. A biochemical identification method based on enzymatic assay was performed, followed by haemolytic and proteolytic tests. PCR-RFLP targeted on the gap gene allowed the genetic identification of strains at the species level and verified phenotypic identification results. A molecular typing method using triplex PCR was performed to recognize the genetic similarity of the analyzed strains. DNA microarray hybridization (StaphyType, Alere Technologies) was used for detection of antibiotic resistance and virulence associated markers. The results obtained indicate high genetic similarity in strains isolated from the same sites. High genetic similarities were also detected between strains isolated from cows from different farms of the same region. A slightly lower similarity was noted however, in strains from various regions indicating that the strains are herd specific and that the cow's infections caused by S. aureus were of a clonal character. In 21 representative isolates selected for DNA-microarray testing, only fosfomycin (fosB) and penicillin resistance markers (blaZ, blaI, blaR) were detected. The presence of genes coding for haemolysins
Genetic methods based on PCR-restriction fragment length polymorphism (RFLP) are widely used for microbial species determination. In this study, we present the application of saoC gene as an effective tool for species determination and within-species diversity analysis for Staphylococcus genus. The unique sequence diversity of saoC allows us to apply four restriction enzymes to obtain RFLP patterns, which appear highly distinctive even among closely related species as well as atypical isolates of environmental origin. Such patterns were successfully obtained for 26 species belonging to Staphylococcus genus. What is more, tracing polymorphisms detected by different restriction enzymes allowed for basic phylogeny analysis for Staphylococcus aureus, which is potentially applicable for other staphylococcal species.
Verotoxin-producing Escherichia coli (VTEC) are nowadays among the most important emerging group of food-borne pathogens (VTEC strains cause gastroenteritis that can be complicated by the hemorrhagic colitis or hemolytic uremic syndrome, HUS). Escherichia coli 026 producing verotoxin 2 was isolated and its identity confirmed by examination of phenotype and genotype; the strain was first described in Slovakia in association with the development of HUS in a 4-year-old girl.
The importance of nosocomial infections caused by coagulase-negative staphylococci is constantly growing. The threat primarily affects immunocompromised patients, the elderly and neonates, particularly after invasive surgery. The problem is fundamentally exacerbated by expanding antibacterial drug resistance. A case report is presented of an 86-year-old patient who underwent a ruptured abdominal aortic aneurysm surgery and developed septicaemia upon surgical wound infection. The causal agent was likely a carbapenem-resistant Klebsiella pneumoniae, however, daptomycin-resistant Staphylococcus pettenkoferi was identified in blood cultures in the absence of daptomycin treatment. To the authors' knowledge, the case study presented is the first published episode of daptomycin-resistant S. pettenkoferi strain.
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