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Streptococcus anginosus (milleri) is a diverse group of gram positive bacteria. Molecular methods to establish relationship between strains are poorly developed. Therefore, main tool to study genetic variability is restriction fragment length polymorphism combined with pulsed field gel electrophoresis (RFLP-PFGE). In this communication, we present optimized protocol for S. anginosus PFGE analysis.
The postsegregational killing system of pSM19035 plasmid consists of the proteins Zeta and Epsilon, a toxin and an antidote, respectively. Zeta mutants were isolated with the use of Bacillus subtilis strain with the zeta gene under control of an inducible promoter integrated into the chromosome. Results of mutant analysis point to the amino terminal part of the Zeta protein as being responsible for the toxicity.
Streptococcus anginosus, Streptococcus intermedius and Streptococcus constellatus form a group of related streptococcal species, namely the Streptococcus Anginosus Group (SAG). The group, previously called “milleri” had been rarely described until 1980/1990 as source of infections. Nowadays SAG bacteria are often described as pathogens causing predominantly purulent infections. The number of infections is highly underestimated, as SAG strains are often classified in the microbiology laboratory as less virulent “viridans streptococci”. Epidemiological situation regarding SAG infections in Poland has been unrecognized, therefore we performed a retrospective analysis of strains isolated between 1996 and 2012. Strains suspected of belonging to SAG were re-identified using an automated biochemical approach (Vitek2) and MALDI-TOF MS. We performed first analysis of antibiotic resistance among SAG strains isolated in Poland using automated methods (Vitek2), disk diffusion tests and E-Tests. We also performed PCR detection of resistance determinants in antibiotic resistant strains. Clonal structure of analyzed strains was evaluated with PFGE and MLVF methods. All three species are difficult to distinguish using automated diagnostic methods and the same is true for automated MIC evaluation. Our analysis revealed SAG strains are rarely isolated in Poland, predominantly from purulent infections. All isolates are very diverse on the genomic level as estimated by PFGE and MLVF analyses. All analyzed strains are sensitive to penicillin, a substantial group of strains is resistant to macrolides and the majority of strains are resistant to tetracycline.
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