Background: Wide use of biocidal agents such as benzalkonium chloride (BCC) and chlorhexidine digluconate (CHX) in hospitals and non-hospital environments, has raised concerns over the emergence of non-susceptible strains. Efflux pumps are of known main mechanisms in biocide tolerance which have been rarely addressed in enterococci - members of gut microbiota which can cause serious problems particularly in hospitalized patients. The purpose of this study was to investigate the susceptibility of enterococci from different sources (clinical and fecal isolates) toward BCC and CHX, and its correlation with efflux associated genes. Also, possible link between biocide tolerance and antibiotic resistance was examined. Materials and Methods: One hundred and four enterococcus isolates including clinical (n = 54) and fecal isolates (n = 50) were studied for susceptibility toward BCC, CHX, ciprofloxacin, gentamicin and vancomycin. Twelve efflux associated genes were investigated by polymerase chain reaction assay. Results: In clinical isolates, reduced susceptibility to CHX and resistance to gentamicin and ciprofloxacin were significantly higher than fecal isolates. Vancomycin resistance was associated with increasing minimum inhibitory concentration of CHX. Among all investigated genes, only three ones, efrA, efrB and emeA were detected which were significantly associated with reduced susceptibility to CHX and were more frequent among clinical isolates. Also, high level resistance to gentamicin was significantly associated with the presence of efrA/B as well as with reduced susceptibility to CHX. Conclusion As expected, reduced susceptibility to CHX, was significantly higher in clinical isolates. However, the presence of a vancomycin-resistant enterococci among fecal isolates of healthy people which showed resistance/tolerance to studied antimicrobial agents, was unexpected and highlights the need to investigate other non-hospital environments to avoid dissemination of antimicrobial resistance. Correlation between reduced susceptibility to CHX and high level resistance to gentamicin, substantiates monitoring of biocide tolerance particularly in the healthcare settings to control the establishment of antimicrobial resistant strains.

BACKGROUND: In many developing countries, shigellosis is endemic and also occurs in epidemics and treatment of multidrug-resistant (MDR) isolates are important. The aims of this study were to determine the antimicrobial susceptibility, prevalence of class 1 and 2 integrons and the clonal relatedness of isolates. MATERIALS AND METHODS: Antimicrobial susceptibility tests were performed by disc diffusion method. Polymerase chain reaction (PCR)-sequencing technique was employed for detection and characterization of integrons. The genetic relatedness was evaluated by using enterobacterial repetitive intergenic consensus (ERIC) PCR. RESULTS: There was a high percentage of resistance to trimethoprim-sulfamethoxazole (TMP/SMX) (93.7%), ampicillin (AMP) (87.3%), streptomycin (STR) (84.5%) and tetracycline (TET) (78.9%). Multidrug resistant phenotype was seen in 95.1% of total isolates. Most common MDR profile was TMP/SMX/STR/AMP resistant pattern. Among the 142 Shigella spp. analyzed in this study, 28 isolates were positive for class 1 integron with two types of gene cassette arrays (dfrA17/aadA5 = 31.7% and dfrA7 = 3.8%). The class 2 integron was more frequently detected among the isolates (94.7%) with dfrA1/sat1/aadA1 (69.4%) and dfrA1/sat1 (30.6%) gene cassettes. ERIC-PCR results showed 6, 5, 4 and 3 main genotypes among S. flexneri, S. sonnei, S. boydii and S. dysenteriae isolates, respectively. CONCLUSIONS: Our findings revealed that multidrug resistant Shigella species with high prevalence of class 2 integron were very common in Iran. In addition, ERIC-PCR patterns showed limited variety of clones are responsible for shigellosis in the region of the study.
Ampicillin ; Clone Cells ; Consensus ; Developing Countries ; Diffusion ; Dysentery, Bacillary ; Genotype ; Integrons ; Iran* ; Methods ; Phenotype ; Polymerase Chain Reaction ; Prevalence ; Shigella* ; Streptomycin ; Tetracycline ; Trimethoprim, Sulfamethoxazole Drug Combination

BACKGROUND: In many developing countries, shigellosis is endemic and also occurs in epidemics and treatment of multidrug-resistant (MDR) isolates are important. The aims of this study were to determine the antimicrobial susceptibility, prevalence of class 1 and 2 integrons and the clonal relatedness of isolates. MATERIALS AND METHODS: Antimicrobial susceptibility tests were performed by disc diffusion method. Polymerase chain reaction (PCR)-sequencing technique was employed for detection and characterization of integrons. The genetic relatedness was evaluated by using enterobacterial repetitive intergenic consensus (ERIC) PCR. RESULTS: There was a high percentage of resistance to trimethoprim-sulfamethoxazole (TMP/SMX) (93.7%), ampicillin (AMP) (87.3%), streptomycin (STR) (84.5%) and tetracycline (TET) (78.9%). Multidrug resistant phenotype was seen in 95.1% of total isolates. Most common MDR profile was TMP/SMX/STR/AMP resistant pattern. Among the 142 Shigella spp. analyzed in this study, 28 isolates were positive for class 1 integron with two types of gene cassette arrays (dfrA17/aadA5 = 31.7% and dfrA7 = 3.8%). The class 2 integron was more frequently detected among the isolates (94.7%) with dfrA1/sat1/aadA1 (69.4%) and dfrA1/sat1 (30.6%) gene cassettes. ERIC-PCR results showed 6, 5, 4 and 3 main genotypes among S. flexneri, S. sonnei, S. boydii and S. dysenteriae isolates, respectively. CONCLUSIONS: Our findings revealed that multidrug resistant Shigella species with high prevalence of class 2 integron were very common in Iran. In addition, ERIC-PCR patterns showed limited variety of clones are responsible for shigellosis in the region of the study.
Ampicillin ; Clone Cells ; Consensus ; Developing Countries ; Diffusion ; Dysentery, Bacillary ; Genotype ; Integrons ; Iran* ; Methods ; Phenotype ; Polymerase Chain Reaction ; Prevalence ; Shigella* ; Streptomycin ; Tetracycline ; Trimethoprim, Sulfamethoxazole Drug Combination

Background: Acinetobacter baumannii is one of the most important hospital pathogenic bacteria that cause infectious diseases. The present study aimed to determine the frequency of carbapenem resistance genes in association with transposable elements and molecular typing of carbapenem-resistant A. baumannii bacteria collected from patients in Shiraz, Iran. Materials and Methods: A total of 170 carbapenem-resistant A. baumannii isolates were obtained from different clinical specimens in two hospitals. The minimum inhibitory concentrations (MIC) of imipenem were determined and the prevalence of OXA Carbapenemases, Metalloβ-lactamases genes, insertion sequences (IS) elements, and transposons were evaluated by the polymerase chain reaction (PCR) method. Finally, molecular typing of the isolates was performed by the Enterobacterial Repetitive Intergenic Consensus-PCR method. Results: The MICs ranged from 16 to 1,024 µg/mL for imipenem-resistant A. baumannii isolates. Out of the 170 carbapenem resistant A. baumannii isolates, bla OXA-24-like (94, 55.3%) followed by bla OXA-23-like (71, 41.7%) were predominant. In addition, A. baumannii isolates carried bla VIM (71, 41.7%), bla GES (32, 18.8%), bla SPM (4, 2.3%), and bla KPC (1, 0.6%). Moreover, ISAba1 (94.2%) and Tn2009 (39.2%) were the most frequent transposable elements. Furthermore, (71, 44.0%) and (161, 94.7%) of the ISAba1 of the isolates were associated with bla OXA-23 and bla OXA-51 genes, respectively. Besides (3, 1.7%), (1, 0.6%) and (5, 2.9%) of bla OXA-23 were associated with IS18, ISAba4, and ISAba2, respectively. Considering an 80.0% cut off, clusters and four singletons were detected. Conclusion According to the results, transposable elements played an important role in the development of resistance genes and resistance to carbapenems. The results also indicated carbapenem-resistant A. baumannii bacteria as a public health concern.