The Role of Prophage ϕSa3 in the Adaption of Staphylococcus aureus ST398 Sublineages from Human to Animal Hosts.

Staphylococcus aureus sequence type (ST) 398 is a lineage affecting both humans and livestock worldwide. However, the mechanisms underlying its clonal evolution are still not clearly elucidated. We applied whole-genome sequencing (WGS) typing to 45 S. aureus strains from China and Canada between 2005 and 2014, in order to gain insight into their evolutionary pathway. Based on WGS phylogenetic analysis, 42 isolates were assigned to the human-associated clade (I/II-GOI) and 3 isolates to livestock-associated clade (IIa). Phylogeny of ϕSa3 sequences revealed five phage groups (Groups 1-5), with Group 1 carrying ϕSa3-Group 1 (ϕSa3-G1), Group 2 carrying ϕSa3-G2, Group 3 carrying ϕSa3-G3, Group 4 carrying ϕSa3-G4 and Group 5 lacking ϕSa3. ϕSa3-G1 was only found in strains that accounted for the most ancestral human clade I, while ϕSa3-G2, ϕSa3-G3 and ϕSa3-G4 were found restricted to sublineages within clade II-GOI. Some isolates of clade II-GOI were also found to be ϕSa3-negative or resistant to methicillin which are unusual characteristics for human-adapted isolates. This study demonstrated a strong association between phylogenetic grouping and phage type, suggesting an important role of ϕSa3 prophage in the evolution of human-adapted ST398 subclones. In addition, our results suggest that this subclone slowly began to adapt to animal hosts by losing ϕSa3 and acquiring methicillin resistance, which was observed in some strains of human-associated clade II-GOI, an intermediate human to livestock transmission clade.

Genetic diversity and methicillin resistance of Staphylococcus aureus originating from buffaloes with mastitis in Iran.

The aims of the present study were to investigate the genetic diversity and methicillin resistance in S. aureus isolates recovered from mastitis-affected buffaloes. Five hundred seventy-eight milk samples were obtained from buffaloes with mastitis in three provinces, Iran. Ninety-one of the 578 tested samples contained S. aureus (15.74%), in two cases were methicillin resistant S. aureus (MRSA). Isolates were typed by spa typing, followed by MLST on some representative isolates and SCCmec typing for MRSA strains. The presence of genes encoding Panton-Valentine leukocidin (PVL) was also tested by PCR. Eight spa types were identified, with t3576 (n = 18), t7311 (n = 18) and t937 (n = 17) were the most common, followed by t304 (n = 11), t7308 (n = 9), t521 (n = 7), t267 (n = 6), and t527 (n = 5). MLST revealed four different sequence types (STs) including ST97 (related to t521 and t527 spa types), ST352 (related to t267), ST291 (related to t304 and t937) and ST522 (related to t7338, t7311 and t3576). Two MRSA were identified as t304-ST291-SCCmecIV and t7311-ST522-SCCmecIV. No PVL-positive S. aureus were found. A significant difference in geographical distribution of genotypes was observed, with some types being prevalent in all studied provinces (P < 0.001). The results demonstrated genetic diversity among the S. aureus strains involved in mastitis in buffaloes. This study also provides evidence of the presence of MRSA belonging to genotypes which have been earlier reported in human infections, emphasizing the need for their epidemiological monitoring.

Antimicrobial effect of Licochalcone A and Epigallocatechin-3-gallate against Salmonella Typhimurium isolated from poultry flocks.

BACKGROUND AND OBJECTIVES: Salmonellosis due to multi-drug resistant Salmonella Typhimurium with biofilm formation ability is a serious public health threat worldwide. Studies have shown that medicinal plants inhibit the growth of bacterial species. The present study aimed at determining antibiotic resistance pattern and biofilm formation ability of S. Typhimurium isolated from poultry flocks. Moreover, the antibacterial activity of Licochalcone A (LAA) and Epigallocatechin-3-gallate (EGCG) against the studied isolates were investigated in this study. MATERIALS AND METHODS: Antibiotic susceptibility testing of S. Typhimurium RITCC1730 and 23 clinical isolates of S. Typhimurium against 8 antibiotics was performed using standard Kirby-Bauer disc diffusion method. The extent of biofilm formation was measured by Microtiter dish biofilm formation assay. Antimicrobials activities of LAA and EGCG were determined by MIC and MBC assays using microdilution method. RESULTS: The highest antimicrobial resistance was detected against chloramphenicol (52.17%), followed by furazolidone (26.08%), and trimethoprim/sulfamethoxazole (21.73%). All isolates were sensitive to ciprofloxacin (100%), followed by gentamicin, imipenem (95.65%), and cefixime (91.30%). Most of the isolates (78.26%) were able to produce weak biofilm. LAA and EGCG inhibited the growth of S. Typhimurium at the MIC levels of 62.5∼1000 and 1.56∼400 µg/mL, respectively. The MBC value of LAA was >1000 µg/mL, while the corresponding value of EGCG varied from 100 to 800 µg/mL. CONCLUSION: S. Typhimurium isolates revealed a multiple antibiotic resistance with biofilm production ability. As a result, EGCG, and to a lesser extent, LAA displayed potential antibacterial activity against S. Typhimurium and could be considered as useful compounds for the development of antibacterial agents against salmonellosis.

Use of 5-Enolpyruvylshikmate-3-Phosphate Synthase Encoding Gene for Typing of Staphylococcus aureus Isolated from Skin and Urinary Tract Infections of Human.

OBJECTIVE(S): Staphylococcus aureus is both a successful human commensal and a major pathogen. In this study we investigated the genetic diversity of 26 S. aureus isolates recovered from human skin and urinary tract infections. MATERIALS AND METHODS: Typing procedure for the studied S. aureus isolates was performed based on PCR amplification of the aroA gene, which encodes the enzyme 5-enolpyruvylshikmate-3-phosphate synthase (EPSPS) that involves in aromatic amino acid biosynthesis, and restriction fragment length polymorphism (RFLP) analysis of the product. RESULTS: All S. aureus isolates produced a single PCR amplification product of 1,153 bp. Digestion of the PCR products with the TaqI endonuclease revealed four different aroA gene patterns designated as A, B, N and H according to the nomenclature system of previous studies. In general, 80.77% of the studied isolates displayed type N, 7.69% were type B, 7.69% were type H and 3.85% displayed type A. CONCLUSION: Divergent aroA types were detected among S. aureus isolates from skin and urinary tract infections. The results showed that urinary tract infections were contaminated by S. aureus isolates with identical banding patterns (A), while isolates recovered from skin infections had different aroA types. This study also indicates that aroA genotypes vary not only from region to region, but also in individual hosts within a region.