The aim of this study was to characterize virulence determinants and antibiotic resistance profiles in enterococci obtained from various clinical sources in the northwest of Iran. A total of 160 enterococcal clinical isolates from various wards of University Teaching Hospitals were collected and specified by biochemical test, from September 2014 to July 2015. Identification of enterococci was confirmed by multiplex PCR in the genus and species level. Antibiotic resistance properties and virulence determinants were examined by phenotypic and molecular methods. Of 160 enterococcal isolates, 125 (78.12%) and 35 (21.88%) isolates were identified as Enterococcus faecalis and Enterococcus faecium, respectively. The most common antibiotic nonsusceptible pattern observed was resistance toward rifampicin [n = 122 (76.25%)] followed by erythromycin [n = 117 (73.12%)]. Among all isolates, gelE [n = 140 (87.5%)], cpd [n = 137 (85.6%)], and asa1 [n = 118 (73.8%)] were the most prevalent virulence genes studied. Thirty isolates (11 E. faecalis, 19 E. faecium) were found to be resistant to vancomycin, with minimum inhibitory concentration of ≥256 μg/ml. Twenty-seven isolates carried the vanA gene, whereas none of the isolates carried vanB. E. faecalis had a considerable ability to show virulence genes and drug resistance. Emergence of antibiotic-resistant enterococci and the high prevalence of virulence traits in our study could be regarded as an alarming situation.

BACKGROUND: Enterococci are naturally found in the gastrointestinal (GI) tract of animals and humans, as well as animal-derived foods and vegetables. We here aimed to determine the prevalence, antibiotic resistance, and virulence determinants of E. faecium and E. faecalis in traditional cheese in the North-west of Iran.MATERIALS AND METHODS: Fifty specimens of popular traditional cheese from dairy stores of Urmia and Tabriz, Iran, were collected. Identification of the genus and species of enterococci was done using molecular and phenotypic techniques.RESULTS: Forty-eight (96 %) of 50 traditional cheese samples were harboring Enterococcus spp, including Enterococcus faecalis (n= 40; 83.33 %) and E. faecium (n= 8; 16.67 %). The prevalence of enterococci ranged from 1.1×105 to 9.7×104 CFU/g, and 1.1×103 to 9.8×103 CFU/g in Urmia and Tabriz samples, respectively. Rifampicin resistance (n= 38; 79.2 %) was the most common pattern observed in the susceptibility test, which was followed by quinupristin/dalfopristin (n= 33; 68.75 %). Among E. faecalis isolates, cpd (100 %), ace (92.5 %) and gelE (87.5 %), and among E. faecium isolates, gelE (100 %) and asa1 (75 %) were found to have the most common virulence genes.CONCLUSION: E. faecalis was the predominant species, displaying more virulence determinants. It also had high antibiotic resistance, as compared to E. faecium. The enterococci identified here commonly expressed virulence and antibiotic resistance determinants. So, it is required to improve the maintenance and production quality of traditional cheese to avoid enterococci contamination.

This study was conducted to investigate the phenotypic and genotypic characteristics of vancomycin-resistant Enterococcus faecalis and Enterococcus faecium. Antibiotic resistance and virulence genes in the aforementioned resistant isolates were studied using the epsilometer (E)-test and polymerase chain reaction (PCR). These isolates were subjected to typing by pulsed-field gel electrophoresis (PFGE). Thirty vancomycin-resistant enterococci (VRE; 18.75%) were isolated from a total of 160 various clinical specimens cultured for any bacterial growth. Of these, 11 (36.7%) isolates were identified as E. faecalis and 19 (63.3%) as E. faecium. Minimum inhibitory concentrations (MICs) of vancomycin, teicoplanin, and three alternative therapeutic options (linezolid, daptomycin, and quinupristin/dalfopristin) were determined using the E-test. Multiplex PCR was done for confirming species, identification of the resistant genotypes, and the detection of the virulence genes. Finally, the clonal relationship of all VRE strains was studied by PFGE. All VRE strains showed vancomycin MIC ≥256 μg/mL, and 27 (90%) isolates carried the vanA gene, whereas none of the isolates carried vanB. The most common resistance antibiotic pattern observed was toward rifampicin (n = 30 [100%]). Among all virulence genes studied, gelE (n = 28 [93.33%]) was found as the most prevalent virulent gene. VRE isolates exhibited 90%, 46.67%, 100%, and 66.67% resistance to teicoplanin, linezolid, quinupristin/dalfopristin, and daptomycin, respectively. Molecular typing demonstrated 16 PFGE types of VRE isolates (A-P). Although vanA was carried by most of the isolates, PFGE displayed small clonal dissemination among VR E. faecium and VR E. faecalis species.

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e247 the circulating HIV-1 subtypes in our country. Moreover, it serves as a useful tool to determine the prevalence and distribution of HIV-1 subtypes and segregation of these subtypes among the different risk groups in Yangon (Lower Myanmar). Forty plasma samples were collected from 3 different risk groups: 10 from heterosexual male, 10 from heterosexual female, 12 from commercial sex workers (CSWs) and 8 from injecting drug users (IDUs) during 2000-2003. The viral RNA from plasma samples of 40 Western-blott confirmed HIVinfected individuals were extracted. These purified viral RNAs were then amplified by Reverse Transcription Polymerase Chain Reaction (RT-PCR) and subsequently by nested (second round) PCR with the primers for gag (p24) region. The reference plasmid DNAs were also amplified by nested PCR with the primers for gag (p24) region. Out of 40 samples, only 22 samples were amplified by gag (p24) PCR allowing subsequent subtype determination. HIV-1 subtype of 22 samples were determined by HIV -1 Gag (p24) HMA. Of the 22 samples, 9 (40.9%) were subtype C, 5 (22.7%) were subtype B' (Thai B cluster within Subtype B), 2 (9.1%) were subtype E (CRF 01-AE), 2 (9.1%) were subtype A and 4 (18.2%) were untypable. Sequencing is normally recommended for these untypeable samples. From this study, HIV-1 gag subtypes C was newly discovered in Yangon (Lower Myanmar) samples for the first time, but previously found in Mandalay (Central Myanmar) since 2000. Subtype C was most commonly found (9 out of 22) in our study and was more predominant in the heterosexual group. In fact, this study is the pioneer ones in Myanmar to apply the HMA technique in our own laboratory setting. The gag subtype A and E was detected only among persons presumably infected sexually and the subtype B' and C were found in both IDUs and persons with sexual exposure. If possible, further confirmation of these subtypes by sequencing should be done. Out of 40 samples non-amplifiable 18 samples were subsequently amplified with second primer set for gag (p 17) region. Only 4 samples amplified but not classified into subtypes due to the limitation of facilities (lack of the corresponding reference plasmids for gag (p 17) region). So, transfer of HIV-1 gag/env HMA technology to the developing countries contributes to more accurate estimations of the prevalence of HIV-1 subtypes and intersubtype recombinants and segregation of these subtypes in different risk groups in those parts of the world where the technology for sequencing is not readily available.