Is There a Real Relationship between the Presence of Helicobacter pylori in Dental Plaque and Gastric Infection? A Genotyping and Restriction Fragment Length Polymorphism Study on Patient Specimens with Dyspepsia in Southwest Iran.

Background: The oral cavity can act as an extra gastric reservoir for H. pylori, and the presence of the bacteria in the oral cavity is associated with a higher risk of dental caries development. This study aimed to determine the genotype and evaluate the association between the presence of H. pylori in dental plaque and gastric biopsy specimens in dyspeptic patients in Ahvaz, Southwest Iran. Methods: In this study, 106 patients with recruited dyspeptic complaints were selected, and from each patient, two gastric antral biopsy specimens and two dental plagues were examined. The presence of H. pylori was identified by the rapid urease test (RUT) and the amplification of ureAB and 16S rRNA genes. Also, to verify a hypothetical mouth-to-stomach infection route, the enzymatic digestions of three genes of cagA, vacA, and ureAB in H. pylori strains isolated from dental plaques and stomach samples were compared for each same case. Results: H. pylori was found in the stomach of 52.8% (56/106) and the dental plaques of 17.9% (19/106) of the studied cases. On the other hand, H. pylori was recognized in the stomach of all 19 cases with oral colonization. Following a combination of restriction fragment lengths 21 polymorphism (RFLP) patterns of these three known genes on stomach and dental plague samples, 14 and 11 unique patterns were seen, respectively. However, for all H. pylori-positive cases (19), the comparison of RLFP patterns of these genes in dental plaque and gastric biopsy specimens was different for the same case. Conclusions: In this study, it seems that there is no significant association between the presence of H. pylori in dental plaque and the stomach of the same case.

Prevalence of Chlamydia trachomatis, Ureaplasma parvum and Mycoplasma genitalium in Infertile Couples and the Effect on Semen Parameters.

Background: Chlamydia trachomatis, Ureaplasma parvum, and Mycoplasma genitalium are common sexually transmitted microorganisms. Our study aimed to determine the prevalence of C. trachomatis, U. parvum, and M. genitalium in infertile and fertile couples and the effect of these microorganisms on semen parameters. Materials and Methods: In this case-control study, samples were collected from 50 infertile couples and 50 fertile couples and were subjected to the routine semen analysis and Polymerase chain reaction (PCR). Results: C. trachomatis and U. parvum were detected in 5 (10%) and 6 (12%) of semen samples from infertile men. Also, out of 50 endocervical swabs from the infertile women, C. trachomatis and M. genitalium were detected in 7(14%) and 4 (8%) of swab specimens, respectively. In the control groups, all of the semen samples and endocervical swabs were negative. Also, in the group of infertile patients infected with C. trachomatis and U. parvum, sperm motility was lower than uninfected infertile men. Conclusions: The results of this study showed that C. trachomatis, U. parvum, and M. genitalium are widespread among the infertile couples in Khuzestan Province (Southwest of Iran). Also, our results showed that these infections can decrease the quality of semen. For the prevention of the consequences of these infections, we suggest a screening program for infertile couples.

Loop mediated isothermal amplification of Clostridioides difficile isolates in gastrointestinal patients.

This study investigated the prevalence of Clostridioides difficile by culture, multiplex polymerase chain reaction (M-PCR), and loop mediated isothermal amplification (LAMP) in patients with suspected C. difficile infections (CDIs). Also, the results of three methods were compared. All stool specimens collected from CDI suspected patients were cultured on selective C. difficile cycloserine-cefoxitin fructose agar and incubated in an anaerobic jar up to 7 days. The bacterial isolates were identified using standard tests. Multiplex-PCR (M-PCR) was performed for detection of tcdA, tcdB, and tpi genes. The LAMP assay was performed to detect the tcdB gene of C. difficile. C. difficile was isolated from 20.0% (n = 10/50) of samples by culture. M-PCR showed that 34.0% (n = 17/50) of the specimens were positive for C. difficile based on the presence of tpi gene. Out of the 17 C. difficile, 13 strains (76.0%) were positive for tcdB gene using M-PCR. However, the LAMP assay showed that 30.0% (15/50) of specimens were positive for the presence of tcdB gene. M-PCR and LAMP methods showed 100.0% sensitivity compared to the culture method. However, the specificity of the LAMP (87.5%) was relatively higher than the M-PCR (82.5%) compared to the culture. Based on the results of this study, the prevalence of toxigenic C. difficile strains was high in suspected CDI patients. So, the differentiation between toxigenic and non-toxigenic strains is necessary. Our data showed that the LAMP assay is a good method for direct detection of toxigenic C. difficile strains from stool specimens.

Detection of OqxAB Efflux Pumps, a Multidrug-Resistant Agent in Bacterial Infection in Patients Referring to Teaching Hospitals in Ahvaz, Southwest of Iran.

Antibiotic resistance mechanisms in Enterobacteriaceae are causative agents of global health problems. Bacterial infections due to multidrug resistance (MDR) may be mediated by the overexpression of efflux pumps. In this study, we investigated the prevalence of oqxA and oqxB genes as two encoding agents of efflux pumps and the determination of antibiotic resistance rate in clinical isolates of Enterobacteriaceae. In this study, 100 Enterobacteriaceae isolates collected from different clinical specimens of infectious patients, such as wounds, urine, blood, discharge, and abscesses except stool, were examined. Identification of the isolates was performed using standard biochemical tests such as TSI, citrate, urea, lysine, SIM, MR-VP, and gas production. The antimicrobial susceptibility test was carried out by the Kirby-Bauer disk diffusion method according to CLSI guidelines, and finally, the oqxA and oqxB genes were detected by the PCR method. Among 100 Enterobacteriaceae isolates, Escherichia coli and Enterobacter gergoviae were the most common isolates with 71% and 20%, respectively. Also, the lowest isolates belonged to Enterobacter cloacae (3%) and Klebsiella pneumoniae (1%). Out of 100 Enterobacteriaceae isolates, 37 isolates (37%) were positive for at least one of oqxA or oqxB genes, while both of these genes were detected among 12% of them. oqxAB genes were detected in 8 cases of 20 (40%) Enterobacter gergoviae and 4 cases of 71 (5.7%) E. coli isolates. The antimicrobial susceptibility test showed that all isolates (100%) were susceptible to imipenem, while the maximum resistance to piperacillin, ceftriaxone, and cefotaxime were 69%, 55%, and 55%, respectively. Also, the results of this study showed that antibiotic resistance in Enterobacteriaceae isolates caused by oqxAB genes is increasing among patients in Iran. Therefore, identification of resistant isolates and antibiotic monitoring programs are essential to prevent the spread of MDR isolates.

Occurrence of the Legionella species in the respiratory samples of patients with pneumonia symptoms from Ahvaz, Iran; first detection of Legionella cherrii.

BACKGROUND: This study aimed to investigate the occurrence of Legionella species in the respiratory samples of patients with pneumonia symptoms from Ahvaz, Iran by culture and the real-time PCR of 23S-5S rRNA gene spacer region. METHODS AND RESULTS: A total of 123 clinical respiratory samples including 63 pleural aspirates, 57 bronchoalveolar lavage (BAL), and 3 sputum were collected from 65 males and 58 females with pneumonia symptoms. All samples were cultured on the Modified Wadowsky-Yee (MWY) agar. The Legionella species was identified by routine bacteriological tests. The presence of the 16S-23S rRNA spacer region gene was investigated by real-time PCR. The Legionella species were differentiated by sequencing of 16S-23S rRNA gene. A total of 2 (1.6%) BAL specimens were positive for Legionella species by culture method. No Legionella spp. were identified in pleural aspirates and sputum samples by the culture method. Using real-time PCR, 9 (7.3%) samples including 6 BAL, 1 sputum, and 2 pleural aspirates were positive for legionella species. These species were detected in 3 (5.2%) females and 6 males (9.2%). The results of sequencing showed that eight species were L. pneumophila while one was L. cherrii. Also, the 2 isolates that were identified by culture method, were confirmed as L. pneumophila by sequencing. CONCLUSIONS: The results showed that using the real-time PCR has a more efficacy for detecting of Legionella species in respiratory samples. Also, L. pneumophila was the most prevalent species circulating in the southwest region of Iran. So, periodic monitoring programs is recommended to prevent epidemics due to this bacterium.

Antimicrobial resistance patterns and their encoding genes among clinical isolates of Acinetobacter baumannii in Ahvaz, Southwest Iran.

Acinetobacter baumannii is one of the most important organisms in nosocomial infections. Antibiotic resistance in this bacterium causes many problems in treating patients. This study aimed to investigate antibiotic resistance patterns and resistance-related, genes in clinical isolates of Acinetobacter baumannii. This descriptive study was conducted on 124 isolates of Acinetobacter baumannii collected from clinical samples in two teaching hospitals in Ahvaz. The antibiotic resistance pattern was determined by disk diffusion. The presence of genes coding for antibiotic resistance was determined using the polymerase chain reaction method. Out of 124 isolates, the highest rate of resistance was observed for rifampin (96.8%). The resistance rate for imipenem, meropenem, colistin, and polymyxin-B were 78.2%, 73.4%, 0.8% and 0.8%, respectively. The distribution of qnrA, qnrB, qnrS, Tet A, TetB, and Sul1genes were 52.6%, 0%, 3.2%, 93.5% 69.2%, and 6.42%, respectively. High prevalence of tetA, tetB, and qnrA genes among Acinetobacter baumannii isolated strains in this study indicate the important role of these genes in multidrug resistance in this bacteria. • Acinetobacter baumannii is an important human pathogen that has attracted the attention of many researchers Antibiotic resistance in this bacterium causes many problems in treating patients. • The resistance rate for imipenem, meropenem, colistin, and polymyxin-B were 78.2%, 73.4%, 0.8% and 0.8%, respectively. The distribution of qnrA, qnrB, qnrS, Tet A, TetB, and Sul1genes were 52.6%, 0%, 3.2%, 93.5% 69.2%, and 6.42%, respectively.

Rough-type and loss of the LPS due to lpx genes deletions are associated with colistin resistance in multidrug-resistant clinical Escherichia coli isolates not harbouring mcr genes.

The emergence of multidrug-resistant Escherichia coli has become a great challenge in treating nosocomial infections. The polymyxin antibiotic colistin is used as a 'last-line' therapy for such strains, but resistance to colistin is increasingly emerging all over the world. In this study, we investigated lipopolysaccharides (LPS) of colistin-resistant isolates and examined mutations in lpx genes in strains not harbouring mcr genes. We examined 351 clinical E. coli isolates with 38 showing reduced susceptibility to colistin. These isolates were collected from different clinical specimens including blood, urine, and wounds, but no stool. After confirmation of the isolates via a BD Phoenix-100 system (Becton Dickinson, USA), we performed antimicrobial susceptibility tests to characterize the resistance pattern of these isolates to different classes of antibiotics, using the disk diffusion test. The Minimum Inhibitory Concentration (MIC) of colistin was determined using E-test strips. The presence of mobile colistin resistance (mcr-1 and mcr-2) genes was tested for all isolates. LPS (including lipid A) were extracted from all isolates and associated lpx genes analyzed by PCR and sequencing. Among the 38 clinical E. coli isolates with reduced susceptibility to colistin, 52% were resistant to colistin. The MICs of colistin ranged from 0.5 µg/ml to ˃256 µg/ml. Within the 20 colistin-resistant strains, six isolates carried the mcr-1 gene, but not mcr-2. Heterologous expression of the mcr-1 gene in susceptible E. coli DH5α increased the MIC of colistin by eight-fold. The remaining 14 isolates, were negative for both mcr genes. Six isolates were further negative for LPS production and five showed rough LPS phenotypes. Here we present evidence that loss of LPS or lipid A-deficiency can lead to colistin-resistance in clinical E. coli isolates not harbouring mcr genes.

Characterization of SCCmec, Spa Types and Multidrug Resistant of Methicillin-Resistant Staphylococcus aureus Isolates in Ahvaz, Iran.

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most pathogens associated with health care. Molecular typing methods are vital for outbreak investigations of MRSA. The aim of this study was characterization of SCCmec, spa types and multidrug resistant of methicillin-resistant Staphylococcus aureus isolates in Ahvaz, Iran. METHODS: A total of 50 MRSA isolates were determined by using the phenotypic method and mecA gene. Antibiotic resistance profile and SCCmec types were screened using disc diffusion method and PCR, respectively. For spa typing of MRSA isolates, two molecular typing methods including the PCR-sequencing and high-resolution melting (HRM) analysis were used. RESULTS: In the present study, the highest sensitivity of MRSA was to vancomycin and linezolid and the lowest to clindamycin. In the MRSA isolates, 22% were XDR and 78% were MDR. SCCmec type III was found commonly among MRSA. Based on PCR-sequencing and HRM results, 10 different spa types were identified. The spa types t037 and t030 were the most common in this study. CONCLUSION: This study emphasizes the spa variation among MRSA isolates, which may be considered as an important criterion when treating staphylococcal infections. Accurate and early detection of MDR, XDR, or even PDR MRSA isolates strains must be commenced by all clinical microbiology laboratories to reduce the menace of antimicrobial resistance.

Isolation and Identification of Legionella spp. in environmental water sources based on macrophage infectivity potentiator (mip) gene sequencing in southwest Iran.

Legionella species are widespread in natural water sources and man-made aqueous environments, as well as fresh-water. The present study was conducted owing to the lack of research regarding the prevalence of Legionella spp in the water sources of Ahvaz city in southwest Iran. In this study the macrophage infectivity potentiator (mip) gene sequencing was used for identification of various Legionella species isolated from different water sources. In this study, 144 water samples were collected and inoculated on the buffered charcoal-yeast extract (BCYE) agar and modified Wadowsky-Yee (MWY) medium. The DNA was extracted from positive cultures. The Legionella species were confirmed by amplifying a 654 bp fragment of the 16S rRNA gene. The mip gene of all isolates were amplified by PCR and purified for sequencing. The mip gene sequences were analyzed by jPHYDIT software version 1. The results showed a 13.9% (20/144) prevalence of Legionella spp. in water sources of Ahvaz city, southwest Iran. Analyzing of the mip gene sequences showed, out of 20 Legionella isolates, 13 isolates (54.1%) were positive for L. pneumophila, 5 isolates (20.8%) were positive for L. worsleinsis, one isolates for each one of L. dumoffi and L. fairfieldensis, (4.1%). According to our research, the occurrence of Legionella spp in water sources could be a hazard for the health systems especially in the hospitals. The regular monitoring of these water sources by health planners may therefore be useful for decreasing the risk for Legionella spp. infections.

First investigation of the presence of SPATE genes in Shigella species isolated from children with diarrhea infection in Ahvaz, southwest Iran.

Background: SPATE (serine protease autotransporters of enterobacteriaceae) genes are considered as a group of the main virulence factors of Shigella species This study aimed to investigate for the first time the distribution of SPATE genes among Shigella spp. isolated from children with diarrhea infection in Ahvaz, Iran. Methodology: In this study, a total of 74 Shigella isolates were collected between August 2016 and June 2017 from feces of children with diarrhea and identified by biochemical and molecular methods for Shigella species. The frequency distribution of the SPATE genes, including pic, pet, sat, sigA and sepA, was evaluated using PCR. The genetic relationship of all isolates was evaluated by enterobacterial repetitive intergenic consensus-PCR. Results: The most common species of Shigella was S. flexneri, followed by S. sonnei and S. boydii. In total, 95.94% of Shigella isolates had at least one of the SPATE genes. The presence of pic, pet, sat, sigA and sepA genes was confirmed among 35.13%, 27%, 47.29%, 58.1% and 39.18% of Shigella isolates, respectively. Of these SPATE genes, the sat and sigA genes were recognized as the most common autotransporters among S. flexneri and S. sonnei isolates, respectively. Also, either S. flexneri or S. sonnei isolates belonging to a same clone type had similar SPATE genes profile. Conclusion: Our results revealed that the high distribution of SPATE genes among Shigella isolates in our region. Hence, this study highlights a need for epidemiological programs to monitor the distribution of SPATE genes locally for prevention from further dissemination of the Shigella isolates harboring them.

The first report of emerging mobilized colistin-resistance (mcr) genes and ERIC-PCR typing in Escherichia coli and Klebsiella pneumoniae clinical isolates in southwest Iran.

Background: The emergence of the plasmid-mediated mcr colistin-resistance gene in bacteria poses a potential threat for treatment of patients, especially when hospitalized. The aims of this study were to search for the presence of mcr-1 and mcr-2 genes among colistin-resistant Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae) isolates from clinical specimens and to determine the fingerprint of strains by enterobacterial repetitive intergenic consensus sequences PCR (ERIC-PCR) method. Methods: In this study, 712 nonduplicate Enterobacteriaceae isolates from clinical specimens were examined. All of the isolates were subcultured on suitable media, and the isolated colonies were identified by standard biochemical tests. Antimicrobial susceptibility test on 7 antibiotics was performed by disk diffusion method, and minimal inhibitory concentration (MIC) of isolates to colistin was determined by the E-test method. These isolates were typed by ERIC-PCR method, and the presence of mcr-1 and mcr-2 genes was investigated by PCR method. Results: Out of 712 nonduplicate Enterobacteriaceae, 470 isolates, including 351 (74.7%) E. coli and 119 (25.3%) K. pneumoniae, were detected. The results of antibiogram tests showed that most of the isolates (81.3%) were resistant to ceftazidime; however, the most susceptibility among of E. coli and K. pneumoniae isolates was observed (81.5%) to colistin. The typing results by ERIC-PCR method showed 36 and 23 fingerprint patterns for colistin-resistant E. coli and K. pneumoniae strains, respectively. Among 64 (13.6%) colistin-phenotypically-resistant Enterobacteriaceae, 8 isolates (1.7%) had mcr-1 gene. These 8 isolates were attributed to E. coli and K. pneumoniae with 6 and 2 isolates, respectively. Whereas no isolates carrying the mcr-2 gene was found. These colistin-resistant isolates displayed colistin MIC values >2 µg/ml in the antibiotic concentration by E-test method. Conclusion: Spreading of Enterobacteriaceae strains harboring plasmid-mediated mcr could fail the colistin-included therapy regimen as the last line of treatment against multidrug-resistant bacterial infections.

Loop-mediated isothermal amplification for detection of Legionella pneumophila in respiratory specimens of hospitalized patients in Ahvaz, southwest Iran.

BACKGROUND: Legionnaires' disease is an important public health problem that can cause substantial mortality and morbidity. Legionnaires' disease-risk estimation may be compromised by uncertainties in Legionella-detection methods. The aim of this study was the detection of L. pneumophila in respiratory specimens of hospitalized patients with respiratory symptoms by culture, PCR, and loop-mediated isothermal amplification (LAMP) methods. METHODS: Sputum and bronchoalveolar lavage samples were obtained from patients with pneumonia admitted to teaching hospitals in Ahvaz, Iran from June 2016 to March 2017. Isolation of Legionella spp. was done by culturing the samples directly onto buffered charcoal-yeast extract and modified Wadowsky-Yee agar medium. Then, PCR and LAMP assays were performed for detection of L. pneumophila via its mip gene in respiratory specimens. RESULTS: A total of 100 respiratory specimens were collected. Our results showed that 1% of the samples were culture positive for Legionella spp., and 3% and 7% of samples were positive for L. pneumophila using the mip gene on PCR and LAMP assays, respectively. CONCLUSION: Legionnaires' disease should be considered in the diagnosis of pulmonary infectious diseases. Also, the LAMP assay is a faster method with higher sensitivity and specificity than conventional methods, such as PCR and culture, for laboratory diagnosis of Legionnaires' disease.

Prevalence and antimicrobial resistance of Shigella species isolated from diarrheal patients in Ahvaz, southwest Iran.

INTRODUCTION: Shigellosis is a significant global human health problem, and Shigella is in charge of almost 165 million cases of this disease annually, of whom 163 million cases are in developing countries and 1.5 million cases are in developed countries. The main aims of the current survey were to identify Shigella spp. isolated from diarrheal patients by conventional biochemical tests, determine the antimicrobial susceptibility profiles by disk diffusion method, and detect the ipaH gene using the PCR assay. METHODS: The bacterial isolates were identified as Shigella spp. by microbiological tests and were serogrouped by the slide agglutination test. Antimicrobial susceptibility testing was performed using the disk diffusion method. PCR was performed to detect the ipaH gene. RESULTS: The Shigella strains were isolated from 522 patients with various diarrhea, including bloody diarrhea (3%), mucoid plus bloody diarrhea (1.9%), mucoid diarrhea (3.2%), and watery diarrhea (3.2%). Overall, 69 (13.2%) isolates were positive for Shigella spp., of which 34 (49.3%) serotypes were identified as Shigella flexneri, 22 (31.9%) serotypes were identified as Shigella sonnei, 9 (13%) serotypes were identified as Shigella boydii, and 4 (5.8%) serotypes were identified as Shigella dysenteriae. Antibiotic susceptibility tests revealed that the highest resistance percentage was related to ampicillin (82%) and trimethoprim-sulfamethoxazole (77%), and ciprofloxacin and ceftriaxone were the best antibiotics against Shigella isolates. CONCLUSION: We concluded that Shigella spp. can be considered as an etiological agent of diarrhea in southwest Iran. Since the drug resistance pattern of Shigella differs geographically and over time within a country, continuous and regular surveillance program is necessary.

Prevalence of enterotoxin-encoding genes among diverse Shigella strains isolated from patients with diarrhea, southwest Iran.

Shigella spp. are a major cause of bacillary dysentery, particularly among children in developing countries such as Iran. This study aimed to investigate the presence of two important Shigella enterotoxins (ShET-1 and ShET-2), encoded by the set and sen genes, respectively, by polymerase chain reaction (PCR) assay among Shigella species isolated from children affected by shigellosis in Ahvaz, southwest of Iran. In this cross-sectional study, from June 2016 to April 2017, altogether 117 Shigella isolates were collected from fecal specimens of children aged <15 years with diarrhea in Ahvaz, southwest Iran. All isolates were identified by standard microbiological and molecular methods. The presence of enterotoxin genes was determined by PCR. The most prevalent isolate was Shigella flexneri (47.9%), followed by Shigella sonnei (41%) and Shigella boydii (11.1%), respectively. Shigella dysenteriae was not detected in patients' samples. The frequencies of set1A, set1B, and sen genes were 5.1% (6/117), 15.4% (18/117), and 76.9% (90/117), respectively. This study provides initial background on the prevalence and distribution of the Shigella enterotoxin genes in Shigella isolates in southwest of Iran. In addition, this study revealed a high prevalence of sen enterotoxin gene in Shigella species.

Molecular detection of vanA and vanB genes among vancomycin-resistant enterococci in ICU-hospitalized patients in Ahvaz in southwest of Iran.

OBJECTIVE: Nosocomial infections due to vancomycin-resistant enterococci (VRE) are known as a source of spreading these bacteria. The aim of this prospective study was molecular detection of vanA and vanB genes among VRE isolated from patients admitted to intensive care units (ICUs) in Ahvaz in southwest of Iran. MATERIALS AND METHODS: Overall, 243 non-duplicate rectal swab specimens were collected from ICU-hospitalized patients in teaching hospitals affiliated to Ahvaz Jundishapur University of Medical Sciences, Iran. The specimens were inoculated on suitable culture media, and isolates were identified by standard biochemical tests. The susceptibility and resistance of enterococci to 10 antibiotics were determined based on the Clinical and Laboratory Standards Institute guidelines. Resistance to vancomycin was phenotypically detected by vancomycin screening test, and the vanA and vanB genes in vancomycin-resistant isolates were amplified by multiplex PCR method. RESULTS: Of 175 specimens containing enterococci, 129 (73.7%) isolates were detected as Enterococcus faecium and Enterococcus faecalis and 46 (26.3%) isolates as Enterococcus spp. The results of susceptibility test showed high rates of resistance to tetracycline, erythromycin, ciprofloxacin, and ampicillin. Moreover, based on this test, out of 129 Enterococcus isolates, 56 (43.4%) were resistant to vancomycin and teicoplanin. Also, among 59 vancomycin-resistant or semi-susceptible isolates, vanA gene was detected in 54 (91.5%) isolates, while none of the isolates had vanB gene. CONCLUSION: According to the results of this study, to prevent the spread of vancomycin-resistant Enterococcus strains, especially in nosocomial infections, the susceptibility of isolates should be determined before vancomycin prescription.

Dissemination of carbapenem-resistant Acinetobacter baumannii in patients with burn injuries.

BACKGROUND: Carbapenem-resistant Acinetobacter baumannii has emerged as an important cause of infection in burn patients. This study aimed to characterize the antimicrobial susceptibility pattern, determine the prevalence of oxacillinase and metallo-beta-lactamase (MBL) genes, and type the A. baumannii isolates obtained from burn patients. METHODS: During a 1-year period, a total of 40 nonduplicated isolates of A. baumannii were obtained from burn patients who were hospitalized in the Taleghani Burn Hospital in Ahvaz, in the southwest of Iran. Testing for antimicrobial susceptibility was carried out by disk diffusion and E-test. To screen MBL production, a double disk synergy and MBL E-test were performed. The presence of blaOXA-23-like, blaOXA-24-like, blaOXA-51-like and blaOXA-58-like, blaVIM, blaIMP and blaSPM, and blaNDM was sought by polymerase chain reaction (PCR). Repetitive extragenic palindromic sequence-based PCR was carried out for determination of isolates clonality. RESULTS: Overall, 92.5% of isolates were carbapenem-resistant. Polymyxin B, colistin, and ampicillin-sulbactam were the most effective agents in vitro, with a susceptibility rate of 100%, 97.5%, and 72.5%, respectively. According to the double disk synergy and E-test, 55.6% and 97.3% of isolates were MBL producers, respectively. Furthermore, 70% of isolates harbored blaOXA-23-like and 20% were positive for blaOXA-24-like. However, no encoding genes were detected for blaVIM, blaIMP and blaSPM, blaNDM, and blaOXA-58-like. Repetitive extragenic palindromic sequence-based PCR revealed that carbapenem-resistant isolates belonged to four clones, including A, B, C, and D; the predominant clones were B and C. CONCLUSION: The rate of carbapenem resistance was high, and it appeared that blaOXA-23-like and blaOXA-24-like contributed to the carbapenem resistance of A. baumannii isolates. This result suggests that the two predominant clones of A. baumannii were spread among burn patients. In order to prevent future dissemination of resistant isolates among burn patients, an effective infection control plan is necessary.

An Investigation of Antibacterial Resistance Patterns Among Acinetobacter baumannii and Pseudomonas aeruginosa Isolates Collected from Intensive Care Units of a University-Affiliated Hospital in Ahvaz, Iran.

BACKGROUND: In recent decades, multidrug-resistant non-fermenting Gram-negative pathogens, particularly Acinetobacter baumannii and Pseudomonas aeruginosa, have been recognized as a major cause of healthcare-associated and nosocomial infections and outbreaks. OBJECTIVES: The aim of this study was to determine the prevalence and pattern of antibiotic resistance in A. baumannii and P. aeruginosa isolates collected from intensive care units (ICUs). METHODS: One hundred fifty-five clinical isolates, including 80 (51.6%) isolates of A. baumannii and 75 (48.4%) isolates of P. aeruginosa, from hospitalized patients in the ICUs of a teaching hospital in Ahvaz, Iran, were collected from January 1 to December 30, 2013. The organisms were identified with conventional bacteriological methods, and antimicrobial susceptibility testing was performed on all isolates in accordance with clinical laboratory and standards institute (CLSI) guidelines. RESULTS: The maximum resistance rates among A. baumannii isolates were observed for ciprofloxacin and trimethoprim-sulfamethoxazole (96.9% and 95.2%, respectively). For P. aeruginosa isolates, the maximum resistance rates were reported for ceftriaxone and trimethoprim-sulfamethoxazole (97.2% and 92.4%, respectively). CONCLUSIONS: The majority of A. baumannii and P. aeruginosa isolates were found to be resistant to commonly recommended antibiotics. Therefore, surveillance of antibiotic consumption and proper antibiotic administration guidelines are essential for preventing major outbreaks in the future.

Characterization of Oxacillinase and Metallo-ß-Lactamas Genes and Molecular Typing of Clinical Isolates of Acinetobacter baumannii in Ahvaz, South-West of Iran.

BACKGROUND: Carbapenem resistant Acinetobacter baumannii is an important nosocomial pathogen associated with a variety of infections. OBJECTIVES: The current study aimed to characterize the antimicrobial susceptibility, analyze the prevalence of oxacillinase and metallo-ß-lactamase (MBL) genes and molecular typing of clinical isolates of A. baumannii. MATERIALS AND METHODS: A total of 124 non-repetitive isolates of A. baumannii were collected from various clinical specimens in two teaching hospitals in Ahvaz, south-west of Iran. Antimicrobial susceptibility test was carried out by disk diffusion method. The minimum inhibitory concentrations (MICs) of imipenem, meropenem, colistin and tigecycline were determined using E-test. To screen for MBL production, double disk synergy (DDs) test and MBL E-test were performed. The presence of bla OXA-23-like, bla OXA-24-like, bla OXA-51-like, bla OXA-58-like, bla VIM, bla IMP and bla SPM genes was assessed by polymerase chain reaction (PCR). To identify clonal relatedness, all isolates were subjected to repetitive sequence-based PCR (rep-PCR). RESULTS: Based on disk diffusion results, the highest rate of resistance was observed in rifampin (96.8%). Colistin and polymyxin-B were the most effective agents in vitro. According to the MIC results, the rate of resistance to imipenem, meropenem, colistin and tigecycline were 78.2%, 73.4%, 0.8% and 0, respectively. Metallo-ß-lactamase production was positive in 42.3% and 79.4% of the isolates by DDs test and E-test, respectively. All isolates (100%) carried bla OXA-51-like gene. According to the results of multiplex PCR, bla OXA-23-like and bla OXA-24-like genes were detected in 85.6% and 6.2% of carbapenem resistant isolates, respectively. No bla OXA-58- like, bla VIM, bla IMP and bla SPM were detected. By rep-PCR, carbapenem resistant isolates were separated into six genotypes (A to F). Genotype A (30.9%) was the most prevalent (P value < 0.001). Genotypes B and C were found in 28.9% and 26.8% of the isolates, respectively. CONCLUSIONS: The rate of carbapenem resistant A. baumannii isolates were high in this study. Since, bla OXA-58-like or MBL genes were not detected, it seems that resistance to carbapenems is related to bla OXA-23-like and bla OXA-24-like. Moreover, bla OXA-23-like was the most prevalent oxacillinase (OXA) gene. Most of the isolates belonged to one of the four dominant genotypes indicating clonal dissemination in the hospitals under study. In order to control the spread of carbapenem-resistant A. baumannii, infection- control strategies are needed.

Survey of CTX-M Gene Frequency in Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae Isolates Using the Combination Disk and PCR Methods in Ahvaz, Iran.

BACKGROUND: A common mechanism of resistance to beta-lactam antibiotics is the production of beta-lactamase by Gram-negative bacteria. Recently, nonderivative extended-spectrum beta-lactamases (ESBLs) from the TEM and SHV enzymes, such as CTX-M, that were related to different geographical regions have been recognized. OBJECTIVES: The aim of this study was to determine the frequency of the CTX-M gene in ESBL-producing Enterobacteriaceae isolates in hospitalized patients in the teaching hospitals of Ahvaz, Iran. METHODS: Enterobacteriaceae isolates from clinical specimens (other than stool), such as wounds, blood, urine, trachea, discharge, and abscess, were collected and examined. All the isolates were identified using standard biochemical tests. The combination test was carried out based on CLSI criteria for the phenotypic detection of ESBL-producing isolates. After DNA extraction, the CTX-M and CTX-M-1 genes were amplified using PCR among phenotypically positive ESBL isolates. RESULTS: Among 240 Enterobacteriaceae isolates, Escherichia coli and Enterobacter were the most common isolates with 171 (71.3%) and 65 (27.1%), respectively. The combination test results also showed that 108 (45%) Enterobacteriaceae isolates were phenotypic ESBL producers, but 104 (96%) isolates were positive for the blaCTX-M gene and 99 (92%) were positive for the blaCTX-M-1 gene according to the PCR method. CONCLUSIONS: The results of this study phenotypically and genotypically confirmed the high frequency of ESBL-producing strains, such as the CTX-M and CTX-M-1 genes, among Enterobacteriaceae isolates in our region. Therefore, use of antibiotic susceptibility testing for the detection of ESBL isolates prior to the prescription of beta-lactam antibiotics is recommended. This could help prevent the spread of bacteria strains that are resistant to beta-lactam antibiotics.

Molecular detection of metallo-ß-lactamase genes, bla IMP-1, bla VIM-2 and bla SPM-1 in imipenem resistant Pseudomonas aeruginosa isolated from clinical specimens in teaching hospitals of Ahvaz, Iran.

BACKGROUND AND OBJECTIVES: Carbapenem resistant Pseudomonas aeruginosa is a serious cause of nosocomial infections. The main purpose of the study is to determine the prevalence rate of imipenem resistant Pseudomonas aeruginosa carrying metallo-ß-lactamase (MBL) genes. MATERIAL AND METHODS: 236 Pseudomonas aeruginosa isolates were collected from teaching hospitals of Ahvaz University of Medical Sciences during a period of 9 months in 2012. These strains were identified using conventional microbiological tests. The susceptibility of isolates to antibiotics were assessed using disk diffusion test. The IMP-EDTA combination disk phenotypic test was performed for detection of MBL producing strains. Finally, polymerase chain reaction (PCR) was performed to detect MBL genes, bla IMP-1, bla VIM-2 and bla SPM-1 in imipenem resistant strains. RESULTS: Out of 236 examined isolates, 122 isolates (51.4%) were resistant to imipenem. The IMP-EDTA combination test showed that among 122 imipenem resistant strains, 110 strains (90%) were phenotipically MBL producers. Additionally, the results of PCR method showed that 2 strains (1.6%) and 67strains (55%) of imipenem resistant Pseudomonas aeruginosa isolates contained bla VIM-2 and bla IMP-1 genes respectively. No SPM-1gene was found in the examined samples. CONCLUSION: Resistance of P. aeruginosa isolates to imipenem due to MBL enzymes is increasing in Ahavaz. Because of clinical significance of this kind of resistance, rapid detection of MBL producing strains and followed by appropriate treatment is necessary to prevent the spreading of these organisms.