Methicillin-Resistant Staphylococcus haemolyticus Displaying Reduced Susceptibility to Vancomycin and High Biofilm-Forming Ability.

BACKGROUND: Staphylococcus haemolyticus is one of the most frequently coagulasenegative staphylococci isolated from healthcare-associated infections, mainly those related to implanted medical devices. OBJECTIVES: This study aimed to determine the antimicrobial susceptibility profile and biofilm forming capacity of S. haemolyticus isolated from bloodstream infections. METHODS: A total of 40 S. haemolyticus isolates were characterized according to their genetic relatedness by repetitive element sequence based-PCR (REP-PCR), antimicrobial susceptibility profile, SCCmec typing, ability to form biofilm on abiotic surface and occurrence of putative genes related to biofilm formation. RESULTS: One S. haemolyticus was susceptible to all antimicrobials. The other isolates (n=39) were resistant to cefoxitin; and among them 34 (87.2%) harbored the mecA gene into the SCCmec type I (5.9%), type III (29.4%), type IV (5.9%) and type V (20.6%); and 38.2% isolates were designated as NT. Apart from cefoxitin, 94.9% of the isolates were resistant to at least four antimicrobial classes, and 32.5% displayed minimal inhibitory concentration (MIC) values higher than 4.0 µg/mL for vancomycin. All isolates formed biofilm on polystyrene surface and were classified as strong biofilm-producers, except for one isolate. All isolates were negative for icaA gene, and the prevalence of the other genes was as follows: atl, 100%; fbp, 92.5%; aap, 90.0%; and bap, 20.0%. CONCLUSION: This study reports a high prevalence of methicillin-resistant S. haemolyticus displaying decreased susceptibility to vancomycin with the ability to form strong biofilms on abiotic surface. The results support the importance of controlling the adequate use of antimicrobials for the treatment of staphylococcal infections.

Nasal Carriage by Staphylococcus aureus among Healthcare Workers and Students Attending a University Hospital in Southern Brazil: Prevalence, Phenotypic, and Molecular Characteristics.

BACKGROUND: Staphylococcus aureus can asymptomatically colonize the human anterior nares and skin, and nasal colonization by this bacterium represents a potential risk for development of invasive infections. The aim of this study was to determine the prevalence of S. aureus nasal carriage among healthcare workers and students attending a university hospital and to characterize the isolates phenotypically and molecularly. METHODS: A cross-sectional study was performed with 324 volunteers. Cultures from nasal samples were obtained and S. aureus isolates were characterized according to their antimicrobial susceptibility profile and four virulence factors-encoding genes. MRSA isolates were characterized regarding their oxacillin/cefoxitin susceptibility, SCCmec, and REP-PCR types. Potential risks for S. aureus and MRSA carriage were analyzed. RESULTS: Of 324 nasal samples, 42.9% were identified as S. aureus, of which 28.8% were MRSA. S. aureus carriers were significantly higher in males and students (OR = 2.898, 95%CI 1.553-5.410); however, no variables were associated with MRSA carriage. All isolates were susceptible to vancomycin and the highest rate of resistance was observed for penicillin (90.6%). All isolates harbored the coa gene, and 97.8%, the icaA gene; 15.8% and 6.5% were positive for tst and lukS-PV/lukF-PV genes, respectively. Among MRSA isolates, 45% carried the mecA gene but were phenotypically susceptible to oxacillin/cefoxitin; two harbored the tst and none had lukS-PV/lukF-PV genes. All MRSAs were distributed into six SCCmec types and type I (62.5%) was the most frequent. REP-PCR typing identified four main clusters among MRSA isolates. CONCLUSION: High prevalence of healthcare workers and students were identified as nasal carriers of S. aureus exhibiting different antimicrobial resistance profiles, including mecA-positive oxacillin-susceptible S. aureus (OS-MRSA) and the presence of virulence-encoding genes. Both cohorts may represent potential sources for the emergence of a successful S. aureus strain highly adapted to the hospital environment.

Comparison of HRM analysis and three REP-PCR genomic fingerprint methods for rapid typing of MRSA at a Brazilian hospital.

INTRODUCTION: Infections caused by multidrug-resistant bacteria are increasingly common and represent a serious problem for public health. Staphylococcus aureus is one of the major agents of infections, and methicillin-resistant S. aureus (MRSA) has spread worldwide. The aim of this study was to phenotypically and genotypically characterize 55 MRSAs isolated in the University Hospital of Londrina, Paraná, Brazil, during 2010. METHODOLOGY: Bacterial isolates were characterized based on their antimicrobial susceptibility profile, biofilm production capacity, and staphylococcal chromosome cassette mec (SCCmec) type. Determination of clonal groups was performed by polymerase chain reaction using the RW3A, JB1, and BOX A1R primers and high-resolution melting (HRM) analysis. RESULTS: The majority of isolates harbored SCCmec type II. SCCmec III, characteristic of the Brazilian endemic clone, was observed in four strains. Only two isolates harbored SCCmec type IV, which is common in community-acquired MRSA strains. Most isolates also showed resistance to more than four of the tested antimicrobials, and 30 isolates exhibited the ability to produce biofilm. DNA polymorphism analysis showed a higher discriminatory power for the JB1 primer, but RW3A revealed several clonal groups of MRSA with similar genotypic and phenotypic characteristics. HRM analysis showed eight different sequence types. CONCLUSIONS: These results are important for epidemiological studies involving MRSA infections.

Molecular and phenotypic characteristics of methicillin-resistant Staphylococcus aureus isolated from hospitalized patients.

INTRODUCTION: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the leading causes of infections acquired in both community and hospital settings. In this study, MRSA isolated from different sources of hospitalized patients was characterized by molecular and phenotypic methods. METHODOLOGY: A total of 123 S. aureus isolates were characterized according to their genetic relatedness by repetitive element sequence based-PCR (REP-PCR), in vitro antimicrobial susceptibility profile, SCCmec typing and presence of seven virulence factor-encoding genes. RESULTS: REP-PCR fingerprinting showed low relatedness between the isolates, and the predominance of one specific lineage or clonal group was not observed. All isolates were susceptible to teicoplanin and linezolide. All isolates were resistant to cefoxitin and penicillin, and the majority were also resistant to one or more other antimicrobials. Fifty isolates (41.7%) were intermediately resistant to vancomycin. Most isolates harbored SCCmec type II (53.7%), followed by type I (22.8%), type IV (8.1%) and type III (1.6%). All isolates harbored at least two virulence factor-encoding genes, and the prevalence was as follows: coa, 100%; icaA, 100%; hla, 13.0%; hlb, 91.1%, hld, 91.1%; lukS-PV and lukF-PV, 2.4%; and tst, 34.1%. A positive association with the presence of hla and SCCmec type II, and tst and SCCmec type I was observed. CONCLUSION: This study showed the high virulence potential of multidrug-resistant MRSA circulating in a teaching hospital. A high prevalence of MRSA showing intermediate vancomycin resistance was also observed, indicating the urgent need to improve strategies for controlling the use of antimicrobials for appropriate management of S. aureus infections.