Characterization of porcine Trueperella pyogenes by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), molecular typing and antimicrobial susceptibility profiling in Sao Paulo State.

Trueperella pyogenes can be found as a commensal or pathogenic bacterium among animals causing a variety of pyogenic infections in several species. The agent appears to act primarily as an opportunistic pathogen but may disseminate and produce metastatic abscesses accompanied or not by mastitis, metritis or pneumonia. In this study, 30 porcine T. pyogenes strains were identified by MALDI-TOF MS and 16S rRNA sequencing and further evaluated in relation to their resistance and genetic profiles through broth microdilution and single enzyme AFLP. All strains were susceptible to ß-lactams, florfenicol, gentamicin, spectinomycin and tiamulin. The highest resistance rates were observed for sulfonamides, tetracyclines and clindamycin. All isolates could be characterized by SE-AFLP presenting more than 80% of similarity, despite their distinct origins. Four genotypes were detected with the segregation of T. pyogenes ATCC 19411 from Brazilian T. pyogenes strains. No correlation between genotypes and isolates origins and susceptibility profile was observed.

Molecular and antimicrobial susceptibility profiling of Streptococcus dysgalactiae isolated from swine.

Streptococcus dysgalactiae subspecies equisimilis and dysgalactiae were isolated from swine clinical specimens. The subspecies equisimilis presented 2 clonal patterns with 85% genetic similarity, whereas subspecies dysgalactiae presented distinct band pattern with less than 80% similarity with equisimilis genotypes. Isolates presented high MIC values to tetracyclines, danofloxacin, spectinomycin, tiamulin, and clindamycin.

Profiling of Leptospira interrogans, L. santarosai, L. meyeri and L. borgpetersenii by SE-AFLP, PFGE and susceptibility testing--a continuous attempt at species and serovar differentiation.

Leptospirosis is a widespread systemic zoonosis, considered as reemerging in certain developing countries. Although the cross agglutinin absorption test is still considered the standard method for Leptospira identification, it presents several disadvantages. The aim of this study was to characterize Leptospira spp. isolated from various hosts by genotyping and broth microdilution susceptibility testing in an attempt to differentiate Leptospira species, serogroups and serovars. Forty-seven isolates were studied. They were previously serotyped, and species confirmation was performed by 16S rRNA sequencing. Single-enzyme amplified fragment length polymorphism (SE-AFLP) and pulsed-field gel electrophoresis (PFGE) analysis enabled the distinction of L. interrogans from L. santarosai, L. meyeri and L. borgpetersenii in two main clusters. Among L. interrogans, it was possible to differentiate into two new clusters the serogroup Icterohaemorrhagiae from the serogroups Canicola and Pomona. L. santarosai isolates presented higher genetic variation than the other species in both techniques. Interestingly, the minimum inhibitory concentration (MIC) cluster analysis also provided Leptospira serogroup differentiation. Further studies are necessary regarding serovar Bananal isolates, as they presented the highest MIC values for most of the antimicrobials tested. All studied techniques successfully distinguished Leptospira species and serogroups. Despite being library-dependent methods, these approaches are less labor intensive and more economically viable, particularly SE-AFLP, and can be implemented in most reference laboratories worldwide to enable faster Leptospira typing.