Prevalence of biofilm formation and vancomycin-resistant genes among Enterococcus faecium isolated from clinical and environmental specimens in Lorestan hospitals.

BACKGROUND AND OBJECTIVES: The antibiotic resistance among Enterococcus faecium strains has increased worldwide. Additionally, biofilm-forming isolates of E. faecium play an important role in human infections. This study was conducted to investigate the prevalence of virulence and antibiotic resistance genes between biofilm-producing and non-biofilm-producing E. faecium strains. MATERIALS AND METHODS: In this study, 228 E. faecium isolates from clinical and environmental specimens were obtained from different wards of hospitals in Lorestan province (Iran). Then, the pattern of antibiotic resistance and minimum inhibitory concentration (MIC) against ß-lactams, glycopeptides, aminoglycosides and other common antibiotics was investigated using disk diffusion and agar dilution methods. Biofilm formation was investigated using polystyrene microtiter plates. PCR assay was conducted for antibiotic resistance and biofilm related genes. Pulse field gel electrophoresis (PFGE) was used to determine the clonal spread of isolates. RESULTS: Most of isolates (78%) were resistant to penicillin, but all were susceptible to linezolid and tigecycline. The biofilm-producing isolates were more resistant to ß-lactams, glycopeptides and aminoglycosides compared to non-biofilm-producing strains. In biofilm-producing isolates, pilA, pilB, efaAfm and esp were the dominant virulence genes and vanA and pbp5 genes were the dominant resistant genes. PFGE analysis exhibited a similar pattern between the clinical and environmental isolates, suggesting the presence of a common origin of the infection by E. faecium. CONCLUSION: The results of the antibiotic resistance, biofilm assay, and PFGE analysis suggest that there is a common clone of persistent and biofilm-producing strains of E. faecium, which could rapidly disseminate in patients and the environment.

Comparison of Biofilm Formation between Methicillin-Resistant and Methicillin-Susceptible Isolates of Staphylococcus aureus.

BACKGROUND: The aim of this study was to compare the biofilm formation and the prevalence of biofilm-associated genes between the isolates of methicillin-resistant (MRSA) and methicillin-susceptible (MSSA) Staphylococcus aureus. METHODS: In total, 209 S. aureus isolates were collected. The antibiotic susceptibility test was conducted using nine antibiotics according to the guidelines of Clinical and Laboratory Standards Institute. Phenotypic biofilm formation was performed with microtiter plate assay. The polymerase chain reaction was employed to detect icaA, icaD, icaB, icaC, clfA, clfB, fnbA, fnbB, fib, cna, eno, ebps, bbp, mecA, and SCCmec types as well as agr group genes with specific primers. RESULTS: Sixty-four (30.62%) isolates were resistant to methicillin, and 54 (83%) MRSA harbored SCCmec III. Furthermore, 122 (58.3%) isolates belonged to agr group I. Twenty-six (36.1%) MRSA and 42 (28.9%) MSSA isolates were strong biofilm producers (no significant difference). The prevalence of icaA, icaD, icaB, and icaC genes in MSSA isolates was 71, 41, 76, and 72%, respectively. The frequency of clfA, clfB, fnbA, fnbB, fib, cna, eno, ebps, and bbp in MSSA was 100, 100, 56, 46, 74, 54, 78, 11, and 1%, respectively. However, in MRSA isolates, the frequency was 97, 97, 64, 51, 76, 56, 79, and 12% with no track of bbp, respectively. CONCLUSION: Statistical difference between MSSA and MRSA regarding biofilm formation and the frequency of all biofilm-encoding genes was not significant. The majority of the S. aureus isolates harbored clfA, clfB, eno, fib, icaA, and icaD genes.

The Microbial Surface Components Recognizing Adhesive Matrix Molecules (MSCRAMMs) Genes among Clinical Isolates of Staphylococcus aureus from Hospitalized Children.

BACKGROUND: Isolates of Staphylococcus aureus express a myriad of adhesive surface proteins that play important role in colonization of the bacteria on nasal and skin surfaces, beginning the process of pathogenesis. The aim of this study was to screen several of the Microbial Surface Components Recognizing Adhesive Matrix Molecules (MSCRAMMs) genes among the isolate of S. aureus from hospitalized children. METHODS: A total of 22 S. aureus isolates were collected from hospitalized children in Tehran from 2012 to 2013. Detection of the mecA and several adhesive surface proteins genes including clfA, B (encoding clumping factors A, B); fnbA, B (encoding finronectin binding proteins A, B); fib (encoding fibrinogen binding protein); eno (encoding laminin binding protein); cna (encoding collagen binding protein); ebps (encoding elastin binding protein) and bbp (encoding bone sialo-protein binding protein), was performed by PCR. RESULTS: The clfAB genes were detected among all the isolates. The prevalence of fnbA, fnbB, fib, eno, cna, ebps and bbp was 63%, 6%, 50%, 59%, 82%, 63%, 9% and 0%, respectively. CONCLUSION: The high prevalence of these genes is important for future plans in vaccine designation. MRSA and MSSA isolates similarly can produce adhesive surface proteins for colonization.

Relationship between adhesin genes and biofilm formation in vancomycin-intermediate Staphylococcus aureus clinical isolates.

The adherence ability and biofilm production are the characteristic of enhanced virulence among isolates of vancomycin-intermediate Staphylococcus aureus (VISA) strains. Although biofilm-forming properties have been well demonstrated in S. aureus, they still remain unclear among the recently emerged types of VISA strains. The aim of this study was to investigate correlations between the distribution of genes encoding staphylococcal microbial surface components which recognise adhesive matrix molecules (MSCRAMMs), the surface protein A (Spa) types, MLST types and the ability of VISA strains to biofilm formation. Microtiter plate assay (Mtp) results showed that all eleven biofilm producer isolates were adherent at various levels. PCR experiments showed that nine MSCRAMM genes, clfA, clfB, fnbA and fib were detected in all of the strains, indicating a high prevalence. The prevalences of other MSCRAMMs and icaABCD genes were found to be variable and not equally distributed among the VISA strains. There was no direct correlation between the distribution of adhesion-related genes and biofilm formation, which indicates that the presence or absence of these genes cannot be employed as an indicator of the ability to biofilm formation. Isolates which belong to the same Spa and ST types showed similar adherence capacities in the Mtp assay, but significant differences were observed between different Spa types. The findings of this study, using quantitative methods, have shown that genotypically different strains of VISA have different capabilities to produce biofilms. This may be caused by a difference in the spa types of VISA isolates or due to their differences in the expression of MSCRAMM and icaABCD genes.