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Background/aim The aim of this study was to investigate the prevalence of virulence genes as well as patterns of antibiotic resistance in cystitis and pyelonephritis uropathogenic Escherichia coli (UPEC) isolates. Materials and methods Two hundred UPEC isolates were collected from hospitalized patients with pyelonephritis (n = 50) and cystitis (n = 150) in Shafa Hospital in Iran. Antimicrobial susceptibility and ESBL production were determined with confirmatory tests. Polymerase chain reaction assay was performed to determine the prevalence of virulence genes in UPEC strains. Results Of a total 200 UPEC isolates, the highest and lowest resistance rates to antibiotics were for cephalexin (74%) and nitrofurantoin (9%), respectively. Of these isolates, 72 (36%) and 128 (64%) strains were ESBL-positive and ESBL-negative, respectively. The frequency of fimH, papC, and hly was 64%, 38%, and 12%, respectively. The most commonly identified virulence gene in ESBL-positive and ESBL-negative strains was fimH 46 (23%) and 86 (43%), respectively. The hlyA gene was more prevalent among patients with pyelonephritis than cystitis. Conclusion The frequency of virulence genes was not significantly different between pyelonephritis and cystitis UPEC strains in the studied patients, but the prevalence rates of hlyA and papC genes were higher among UPEC strains isolated from inpatients compared to outpatients; hence, they could be considered as useful targets for prophylactic interventions.

Background/aim: Clostridium difficile is a frequent cause of nosocomial infections and has become a major public health concern in developed nations. In the present study, the prevalence and antimicrobial susceptibility pattern of toxigenic C. difficile strains isolated in Iran were investigated.Materials and methods: Between June 2016 and May 2017, 2947 inpatient fecal samples were taken from symptomatic adult hospitalized patients in different units of 32 care facilities in Tehran, Iran. C. difficile strains were identified by microbiological/biochemical methods. Susceptibility to 20 antimicrobials was measured by E-test method. Toxin-specific immunoassays and cytotoxicity assays were used to determine in vitro toxin production.Results: Out of 2947 fecal samples, 538 (18.25%) C. difficile isolates were obtained among those with suspected CDI. In E-test method, all C. difficile isolates were susceptible to fidaxomicin, vancomycin, amoxicillin/clavulanate, and meropenem and were resistant to penicillin G. The prevalence of multidrug resistant C. difficile was 69.33% (373/538). Among 538 C. difficile, 147 (27.32%), 169 (31.41%), and 222 (41.26%) isolates were TcdA+/TcdB+, TcdA-/TcdB+, and TcdA-/TcdB-, respectively.Conclusion: The results evidently support the hypothesis of a probable role of toxigenic strains of C. difficile in developing gastrointestinal complaints in patients with diarrhea.

There are challenges regarding increased global rates of microbial resistance and the emergence of new mechanisms that result in microorganisms becoming resistant to antimicrobial drugs. Fosfomycin is a broad-spectrum bactericidal antibiotic effective against Gram-negative and certain Gram-positive bacteria, such as Staphylococci, that interfere with cell wall synthesis. During the last 40 years, fosfomycin has been evaluated in a wide range of applications and fields. Although numerous studies have been done in this area, there remains limited information regarding the prevalence of resistance. Therefore, in this review, we focus on the available data concerning the mechanisms and increasing resistance regarding fosfomycin.

INTRODUCTION: The Helicobacter pylori infection and cytokine-mediated inflammatory responses play significant roles in the pathogenesis of gastric cancer (GC). This study was performed to determine the association between the risk of GC and genetic polymorphisms in interleukin (IL)-1β and tumor necrosis factor-alpha (TNF-α). METHODS: The polymorphisms of IL-1β and TNF-α genes were analyzed by polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) in 290 patients who underwent endoscopy. Infection with H. pylori was diagnosed by histological analysis, rapid urease test, and PCR of gastric biopsy samples. Quantitative real-time PCR was performed to determine the relative mRNA expression levels. RESULTS: No significant difference was detected in allele frequency and genotype of all studied polymorphisms between chronic gastritis (CG), GC and healthy individuals. IL-1β mRNA was down-regulated in both gastritis (relative quantification (RQ)=0.447) and the GC groups (RQ=0.151). In contrast, the expression of TNF-α was up-regulated in the GC group (RQ=2.817) compared to the gastritis group (RQ=0.861). CONCLUSIONS: The studied single-nucleotide polymorphisms are not risk factors for development of CG and GC. However, H. pylori infection causes a huge increase in the TNF-α expression in GC patients.

Transfusion transmissible infections (TTIs) have been a public health challenge for the accessibility, quality and safety of blood transfusion. The present study aimed to consider the prevalence and the trends of hepatitis B virus (HBV), hepatitis C virus (HCV), Human T-cell leukemia virus type 1 (HTLV-1), human immunodeficiency virus (HIV) and syphilis across the ten years among retrospective blood donors. A retrospective investigation of blood donors' data covering the period from 22 May 2009 to 22 May 2019 was done. Data was accumulated and analyzed from Blood Transfusion Center records, pertaining to all donors who were screened for various TTIs using respective immunological techniques. Out of the 682,171 screened donors in the 2009-2019 study period, 2470 (0.36 %) were infected with at least one infectious agent. The overall prevalence of HBV, HCV, HTLV-1, HIV and syphilis were 1700 (0.25 %), 184 (0.027 %), 335 (0.05 %), 4 (0.0.05 %) and 247 (0.036 %), respectively. The study showed male dominated donor pool (96.79 %) with higher prevalence (0.34 %) of TTIs compared to female donors (0.02 %) with 3.21 % population. Despite the low prevalence of TTIs in our study, HBV, HCV, syphilis and HIV have remained a big threat to safe blood transfusion in Iran. Strict adherence to selection criteria, algorithm of donor screening, use of highly sensitive and specific methods for detection of TTIs, regular consultation and health education programs, prevention and sanitization strategies to reduce the risk of TTIs are recommended to reduce the risk of TTIs and ensure the safety of blood transfusion for recipient.

Background: It is of great importance to quickly and accurately detect Brucella abortus and Brucella melitensis from clinical and non-clinical samples because of their high prevalence and high risk in causing brucellosis, a life-threating infectious disease affecting both humans and animals. Methods: The current study describes a new method for the detection of brucellosis in clinical samples using real-time polymerase chain reaction (PCR) and high-resolution melt (HRM) curve analysis. This study was conducted on 70 human and 55 animal isolates with more than 1/80 serum antibody titers. Additionally, the accuracy and specificity of the methods were compared. Results: The mean range [cycles threshold±standard deviation (CT±SD)] for the amplified samples was 15.39–25.15 by real-time PCR. The melting peak range (°C) ±SD for B. abortus and B. melitensis was 90.10±0.4 and 89.70±0.4, respectively, and 10 was reported on peak height. Conclusions: The results of HRM analysis can be used for species differentiation and bacterial genotyping according to nucleotide polymorphism. This molecular method could help in diagnosing Brucella quickly and precisely. Quick recognition of Brucella species could decrease its prevalence among humans and animals and mitigate economic loss.

It is known that type 2 diabetes (T2D) in humans could be linked to the composition of gut microbiota. The aim of this study was to evaluate three faecal bacterial species, including Bacteroides fragilis, Bifidobacterium longum and Faecalibacterium prausnitzii in patients with T2D. This case control study included 18 patients with T2D and 18 matched persons without diabetes. The concentrations of B. fragilis, B. longum and F. prausnitzii were determined by quantitative Real-Time PCR. Quantitative PCR analysis revealed that the gut bacterial composition in patients with T2D was partially different from that in the healthy individuals. Faecalibacterium prausnitzii was significantly lower in patients with T2D (P-value = 0.038). Bacteroides fragilis was under-represented in the microbiota of the group with diabetes, but its difference between two groups was not significant (P-value = 0.38). No difference was observed for B. longum community between the both groups (P-value = 0.99). Characterization of specific species of intestinal microbiota shows some compositional changes in patients with T2D. The results may be valuable for developing strategies to control type 2 diabetes by modifying the intestinal microbiota. Long-term studies with emphasis on other bacterial groups are suggested to clarify the association of T2D with gut microbiota.

Acinetobacter baumannii is an important opportunistic pathogen that causes major public health concern especially in hospitalized patients due to the acquisition of multidrug resistance (MDR). The aim of this study was to systematically review published data about the prevalence rate of MDR-A. baumannii (MDR-AB) from different parts of Iran and provide an overall relative frequency (RF) using meta-analysis. All available national and international databanks were searched to find published studies up to June 2016. Quality of studies was assessed by STROB and PRISMA forms. Because of the significant heterogeneity observed, random effects model was used to combine the results. STATA SE version 11.2 was used for statistical analysis. Out of the 9646 results, 37 suitable articles were extracted according to inclusion and exlusion criteria. The pooled prevalence of MDR-AB was estimated 72% annually. Relative frequency of MDR-AB in different studies varied from 22.8 to 100%. Since the prevalence of MDR-AB is higher than many other countries, measures should be taken to keep the emergence and transmission of these strains to a minimum.

Aim: Hepatitis E is a form of acute viral hepatitis, which is caused by hepatitis E virus (HEV). The study objective was to evaluate the prevalence of HEV to provide an overall relative frequency in Iran. Methods: In this study, databases such as PubMed and Scopus were searched using the following keywords according to MeSH, “Hepatitis E virus”, “Epidemiology”, “Enzyme-linked immunosorbent assay (ELISA)” and “Iran”. Then studies which had reported prevalence of HEV were selected. Results: We selected 36 researches out of 9246 for our study. From all the samples, the pooled estimated prevalence of HEV positive was 9.7%. It should be noted that HEV infection's relative frequency varied from 1.1% in Tehran to 46.1% in Ahvaz. Conclusion: HEV is common in Iran, although comparing to some neighboring countries, its prevalence is lower.

Urinary tract infection (UTI) is one of the most common infections in humans. It is primarily caused by uropathogenic Escherichia coli (UPEC), which has a high multidrug resistance (MDR). In consideration of the prevalence of MDR-UPEC strains, the aims of the present study were to systematically review the published data about the prevalence rate of MDR-UPEC from different parts of Iran and to establish the overall relative frequency (RF) of these strains in Iran. We searched several databases including PubMed, ISI Web of Science, Scopus, Google Scholar, IranMedex, and Iranian Scientific Information Database by using the following keywords: "Escherichia coli", "multidrug resistant", "MDR", "urinary tract infections", "UTI", "uropathogenic". and "Iran". Articles or abstracts that reported the prevalence of MDR-UPEC were included in this review. We found 15 articles suitable for inclusion in this study. A pooled estimation of 10,247 UPEC strains showed that 49.4% (95% confidence interval = 48.0-50.7%) of the stranis were MDR positive. The RF of MDR-UPEC in different studies varied from 10.5% to 79.2% in the Kashan and Hamedan provinces, respectively. According to the results of the present study, the RF of MDR-UPEC in Iran is high. Thus, measures should be taken to keep the emergence and transmission of these strains to a minimum.

Successful treatment of cancer remains a challenge, due to the unique pathophysiology of solid tumors, and the predictable emergence of resistance. Traditional methods for cancer therapy including radiotherapy, chemotherapy, and immunotherapy all have their own limitations. A novel approach is bacteriotherapy, either used alone, or in combination with conventional methods, has shown a positive effect on regression of tumors and inhibition of metastasis. Bacteria‐assisted tumor‐targeted therapy used as therapeutic/gene/drug delivery vehicles has great promise in the treatment of tumors. The use of bacteria only, or in combination with conventional methods was found to be effective in some experimental models of cancer (tumor regression and increased survival rate). In this article, we reviewed the major advantages, challenges, and prospective directions for combinations of bacteria with conventional methods for tumor therapy.

Background/aim Clostridium difficile is a frequent cause of nosocomial infections and has become a major public health concern in developed nations. In the present study, the prevalence and antimicrobial susceptibility pattern of toxigenic C. difficile strains isolated in Iran were investigated. Materials and methods Between June 2016 and May 2017, 2947 inpatient fecal samples were taken from symptomatic adult hospitalized patients in different units of 32 care facilities in Tehran, Iran. C. difficile strains were identified by microbiological/biochemical methods. Susceptibility to 20 antimicrobials was measured by E-test method. Toxin-specific immunoassays and cytotoxicity assays were used to determine in vitro toxin production. Results Out of 2947 fecal samples, 538 (18.25%) C. difficile isolates were obtained among those with suspected CDI. In E-test method, all C. difficile isolates were susceptible to fidaxomicin, vancomycin, amoxicillin/clavulanate, and meropenem and were resistant to penicillin G. The prevalence of multidrug resistant C. difficile was 69.33% (373/538). Among 538 C. difficile, 147 (27.32%), 169 (31.41%), and 222 (41.26%) isolates were TcdA+/TcdB+, TcdA-/TcdB+, and TcdA-/TcdB-, respectively. Conclusion The results evidently support the hypothesis of a probable role of toxigenic strains of C. difficile in developing gastrointestinal complaints in patients with diarrhea.

Background and objectives:blaSHV, blaTEM and blaVEB are a group of Extended-Spectrum Beta-Lactamase enzymes (ESBLs) which are able to hydrolyze Penicillins and some cephalosporin antibiotics. The present study evaluated the frequency of ESBL genes blaSHV, blaTEM and blaVEB in Acinetobacter baumannii strains isolated from nosocomial infections to outline the importance of these genes in antibiotic resistance. Methods: One hundred Acinetobacter baumannii strains were isolated from different nosocomial infections. After antibiotic resistance evaluation with the Kirby-Bauer disc-diffusion method, the Minimum Inhibitory Concentration (MIC) of Ciprofloxacin was measured using the E-test method. Then, the ESBL producing strains were identified employing Combined Disk Methods. Finally, all isolates were evaluated with the Polymerase Chain Reaction (PCR) technique to detect the ESBL genes of interest. Results: Out of 100 Acinetobacter baumannii isolates, 59% were ESBL positive according to the phenotypic method. The PCR assay could not detect the blaSHV and blaVEB genes in the studied isolates, but the presence of blaTEM gene was demonstrated in 42% of the strains. Conclusion: The high resistance to most antibiotics, the high prevalence of ESBLs-producing strains and also a high prevalence of blaTEM gene in A. baumannii strains found in the current study gives cause for major concern about nosocomial infections in Iran because of the treatment complexity of these strains. Our results highlight the need for infection control measures to prevent the spread of resistant isolates, especially in hospitals., Hintergrund:blaSHV, blaTEM und blaVEB gehören zur Gruppe der Extended-Spectrum Beta-Lactamase Enzyme (ESBLs) codierenden Gene, die Penicilline und einige Cephalosporine hydrolysieren können. In der vorliegenden Studie sollte die Häufigkeit dieser ESBL Gene bei Acinetobacter baumannii Stämmen, isoliert aus nosokomialen Infektionen, untersucht werden. Methode: 100 Acinetobacter baumannii Stämme wurden bei verschiedenen nosokomialen Infektionen isoliert. Nach der Resistenzbestimmung gemäß der Agardiffusionsmethode nach Kirby Bauer wurde die Minimale Hemmkonzentration (MHK) von Ciprofloxacin mit der E-Test Methode bestimmt. Anschließend wurden die ESBL bildenden Stämme mit der Combination Disk-Methode identifiziert und abschließend die Gene mittels PCD detektiert. Ergebnisse: 59% der Isolate waren phänotypisch ESVL positiv. Mittels PCR konnte nur das blaTEM Gen detektiert werden. Schlussfolgerung: Die Resistenz gegen die meisten Antibiotika, die hohe Prävalenz von ESBL Bildnern und des blaTEM Gens bei Acinetobacter baumannii Stämmen ist besorgniserregend in Bezug auf die Behandlung nosokomialer Infektionen im Iran. Die Ergebnisse unterstreichen die Notwendigkeit von Infektionskontrollmaßnahmen zur Verhinderung der Ausbreitung resistenter Stämme vor allem in Krankenhäusern., GMS Hygiene and Infection Control; 14:Doc02

OBJECTIVE To study on antibiotic susceptibility and identify coagulase-negative Staphylococcus (CoNS) species based on tuf gene sequencing from keratitis followed by using soft contact lenses in Isfahan, Iran, 2013. METHODS This study examined 77 keratitis cases. The samples were cultured and the isolation of CoNS was done by phenotypic tests, and in vitro sensitivity testing was done by Kirby-Bauer disk diffusion susceptibility method. RESULTS Thirty-eight of isolates were conveniently identified as CoNS. In this study, 27 (71.1%), 21 (55.3%), and 16 (42.1%) were resistant to penicillin, erythromycin, and tetracycline, respectively. One hundred percent of isolates were sensitive to gentamicin, and 36 (94.7%) and 33 (86.8%) of isolates were sensitive to chloramphenicol and ciprofloxacin, respectively. Also, resistances to cefoxitin were 7 (18.4%). Analysis of tuf gene proved to be discriminative and sensitive in which all the isolates were identified with 99.0% similarity to reference strains, and Staphylococcus epidermidis had the highest prevalence among other species. CONCLUSIONS Results of this study showed that CoNS are the most common agents causing contact lens-associated microbial keratitis, and the tuf gene sequencing analysis is a reliable method for distinguishing CoNS species. Also gentamycin, chloramphenicol, and ciprofloxacin are more effective than the other antibacterial agents against these types of bacteria.

Recent studies indicate that inflammatory reactions leading to the development of type 2 diabetes mellitus (T2DM) may also contribute to variations in the composition of the intestinal microbiota, suggesting a relation between T2DM and bacterial residents in the intestinal tract. This case-control study was designed to evaluate the composition of the gut microbiota dominant bacterial groups in patients with T2DM compared to the healthy people. A total of 36 adult subjects (18 patients diagnosed with T2DM and 18 healthy persons) were included in the study. The intestinal microbiota composition was investigated by quantitative real-time polymerase chain reaction (qPCR) method using bacterial 16S rRNA gene. The quantities of two groups of bacteria were meaningfully different among T2DM patients and healthy individuals. While, the level of Lactobacillus was significantly higher in the patients with T2DM (P value < 0.001), Bifidobacterium was significantly more frequent in the healthy people (P value < 0.001). The quantities of Prevotella (P value = 0.0.08) and Fusobacterium (P value = 0.99) genera in faecal samples were not significantly different between the two groups. The significant alterations in dominant faecal bacterial genera found in T2DM patients participating in the current study highlight the link between T2DM disease and compositional variation in intestinal flora. These findings may be valuable for developing approaches to control T2DM by modifying the gut microbiota. More investigations with focus on various taxonomic levels (family, genus and species) of bacteria are necessary to clarify the exact relevance of changes in the gut microbial communities with the progression of T2DM disorder.

Urinary tract infection(UTI) caused by Gram-negative bacteria is the second most common infectious presentation in community medical practice. Approximately 150 million people are diagnosed with UTI each year worldwide. Drug resistance in Gram-negative uropathogens is a major global concern which can lead to poor clinical outcomes including treatment failure, development of bacteremia, requirement for intravenous therapy, hospitalization, and extended length of hospital stay. The mechanisms of drug resistance in these bacteria are important due to they are often not identified by routine susceptibility tests and have an exceptional potential for outbreaks. Treatment of UTIs depends on the access to effective drugs, which is now threatened by antibiotic resistant Gram-negative uropathogens. Although several effective antibiotics with activity against highly resistant Gram-negatives are available, there is not a unique antibiotic with activity against the high variety of resistance. Therefore, antimicrobial susceptibility tests, correlation between clinicians and laboratories, development of more rapid diagnostic methods, and continuous monitoring of drug resistance are urgent priorities. In this review, we will discuss about the current global status of drug-resistant Gram-negative uropathogens and their mechanisms of drug resistance to provide new insights into their treatment options.

INTRODUCTION: Infections caused by β-lactamase-producing gram-negative bacteria, such as Klebsiella pneumoniae, are increasing globally with high morbidity and mortality. The aim of the current study was to determine antimicrobial susceptibility patterns and the prevalence of antibiotic resistance genes (β-lactamase and integron genes) using multiplex PCR. METHODS One-hundred K. pneumoniae isolates were collected from different clinical samples. Antibiotic susceptibility testing was performed with thirteen different antibiotics. Multiplex-PCR was used to detect β-lactamase (bla TEM, bla CTX-M, bla SHV , bla VEB, bla PER, bla GES, bla VIM, bla IMP, bla OXA, and bla KPC) and integron genes (int I, int II, and int III). RESULTS: The highest and lowest rate of resistance was exhibited against amikacin (93%) and imipenem (8%), respectively. The frequency of β-lactamase-positive K. pneumoniae was 37%, and the prevalence of the bla TEM, bla CTX-M, bla SHV , bla VEB, bla PER, bla GES, bla VIM, bla IMP, bla OXA, and bla KPC genes was 38%, 24%, 19%, 12%, 6%, 11%, 33%, 0%, 28%, and 23%, respectively. Of the 100 isolates, eight (8%) were positive for class I integrons; however, class II and III integrons were not detected in any of the strains. CONCLUSIONS: These results indicate co-carriage of a number of β-lactamase genes and antibiotic resistance integrons on the same plasmids harboring multi-drug resistance genes. It seems that these properties help to decrease treatment complications due to resistant bacterial infections by rapid detection, infection-control programs and prevention of transmission of drug resistance.

Brucellosis is still one of the most challenging issues for health and the economy in many developing countries such as Iran. Considering the high prevalence of brucellosis, the aim of the current study was to systematically review published data about the annual incidence rate of this infection from different parts of Iran and provide an overall relative frequency (RF) for Iran using meta-analysis. We searched several databases including PubMed, ISI Web of Science, Scopus, google scholar, IranMedex and Iranian Scientific Information Database (SID) by using the following keywords: “Brucella”, “Brucellosis”, “Malta fever”, “Mediterranean fever”, “undulant fever”, “zoonosis” and “Iran” in Title/Abstract/Keywords fields. Articles/Abstracts, which used clinical specimens and reported the incidence of brucellosis, were included in this review. Quality of studies was assessed by STROB and PRISMA forms. All statistical analyses were performed using STATA 11.0 (STATA Corp, College Station, TX) and P-values under 0.05 were considered statistically significant. Out of the 8326 results, we found 34 articles suitable, according to inclusion and exlusion criteria, for inclusion in this systematic review and meta-analysis. The pooled incidence of brucellosis was estimated 0.001% (95% confidence interval (CI) = 0.0005–0.0015%) annually. Relative frequency of brucellosis in different studies varied from 7.0/100000 to 276.41/100000 in Qom and Kermanshah provinces, respectively. This systematic-review and meta-analysis study showed that the highest incidences of brucellosis are occurred in west and northwest regions of Iran. Totally, the incidence of the disease in Iran is in the high range.

Brucella is zoonotic pathogen that induces abortion and sterility in domestic mammals and chronic infections in humans called Malta fever. It is a facultative intracellular potential pathogen with high infectivity. The virulence of Brucella is dependent upon its potential virulence factors such as enzymes and cell envelope associated virulence genes. The aim of this study was to investigate the Brucella virulence factors among strains isolated from humans and animals in different parts of Iran. Seventy eight strains of Brucella species isolated from suspected human and animal cases from several provinces of Iran during 2015–2016 and identified by phenotypic and molecular methods. The multiplex-PCR (M-PCR) assay was performed in order to detect the ure, wbkA, omp19, mviN, manA and perA genes by using gene specific primers. Out of 78 isolates of Brucella spp., 57 (73%) and 21 (27%) isolates were detected as B. melitensis and B. abortus, respectively, by molecular method. The relative frequency of virulence genes ure, wbkA, omp19, mviN, manA and perA were 74.4%, 89.7%, 93.6%, 94.9%, 100% and 92.3%, respectively. Our results indicate that the most of Brucella strains isolated from this region possess high percent of virulence factor genes (ure, wbkA, omp19, mviN, manA and perA) in their genome. So, each step of infection can be mediated by a number of virulence factors and each strain may have a unique combination of these factors that affected the rate of bacterial pathogenesis.

Brucellosis is one of the most common zoonotic diseases of animal and human beings. This study aimed to differentiate the Brucella spp. and determines the patterns of biovars by using repetitive element palindromic (REP)-PCR and PCR restriction fragment length polymorphism (RFLP) methods. A total of 100 blood specimens suspected of harbouring brucellosis were collected. Conventional culture methods and multiplex PCR were used for the detection of Brucella genus and species; and REP-PCR was used for Brucella spp. differentiation and polymorphisms sequence analysis. In addition, to identify the biovar patterns of REP-PCR, PCR-RFLP was used. Eighty-three samples were identified as harbouring Brucella spp. by the implementation of multiplex PCR, 72 of which were detected as Brucella melitensis and 11 as B. abortus. Also, through analysing the results of PCR-RFLP, it was found that of 72 B. melitensis samples, 69 were B. melitensis biovar 1 and three species were from other biovars. In addition, the obtained patterns for all of the B. abortus samples were from biovars 3, 5, 6 and 9. This study also optimized a test for the detection of Brucella biovar with the REP-PCR method such that Brucella spp. and biovars could be separated in the shortest possible time. Keywords: Brucella abortus, B. melitensis, Polymorphisms, REP-PCR, PCR-RFLP

Background Taxonomic and phylogenetic studies of Mycobacterium species have been based around the 16sRNA gene for many years. However, due to the high strain similarity between species in the Mycobacterium genus (94.3% - 100%), defining a valid phylogenetic tree is difficult; consequently, its use in estimating the boundaries between species is limited. The sequence of the rpoB gene makes it an appropriate gene for phylogenetic analysis, especially in bacteria with limited variation. Objectives In the present study, a 360bp sequence of rpoB was used for precise classification of Mycobacterium strains isolated in Isfahan, Iran. Materials and methods From February to October 2013, 57 clinical and environmental isolates were collected, subcultured, and identified by phenotypic methods. After DNA extraction, a 360bp fragment was PCR-amplified and sequenced. The phylogenetic tree was constructed based on consensus sequence data, using MEGA5 software. Results Slow and fast-growing groups of the Mycobacterium strains were clearly differentiated based on the constructed tree of 56 common Mycobacterium isolates. Each species with a unique title in the tree was identified; in total, 13 nods with a bootstrap value of over 50% were supported. Among the slow-growing group was Mycobacterium kansasii, with M. tuberculosis in a cluster with a bootstrap value of 98% and M. gordonae in another cluster with a bootstrap value of 90%. In the fast-growing group, one cluster with a bootstrap value of 89% was defined, including all fast-growing members present in this study. Conclusions The results suggest that only the application of the rpoB gene sequence is sufficient for taxonomic categorization and definition of a new Mycobacterium species, due to its high resolution power and proper variation in its sequence (85% - 100%); the resulting tree has high validity.

Urinary tract infection (UTI) is one of the most common infections in humans. It is primarily caused by uropathogenic Escherichia coli (UPEC), which has a high multidrug resistance (MDR). In consideration of the prevalence of MDR-UPEC strains, the aims of the present study were to systematically review the published data about the prevalence rate of MDR-UPEC from different parts of Iran and to establish the overall relative frequency (RF) of these strains in Iran. We searched several databases including PubMed, ISI Web of Science, Scopus, Google Scholar, IranMedex, and Iranian Scientific Information Database by using the following keywords: "Escherichia coli", "multidrug resistant", "MDR", "urinary tract infections", "UTI", "uropathogenic". and "Iran". Articles or abstracts that reported the prevalence of MDR-UPEC were included in this review. We found 15 articles suitable for inclusion in this study. A pooled estimation of 10,247 UPEC strains showed that 49.4% (95% confidence interval = 48.0-50.7%) of the stranis were MDR positive. The RF of MDR-UPEC in different studies varied from 10.5% to 79.2% in the Kashan and Hamedan provinces, respectively. According to the results of the present study, the RF of MDR-UPEC in Iran is high. Thus, measures should be taken to keep the emergence and transmission of these strains to a minimum.

Background: Pseudomonas aeruginosa is a well-known opportunistic pathogen, which affects hospitalized patients in different wards due to its natural resistance to drugs. Objectives: The purpose of the current study was to determine the antibiotic susceptibility profiles and genetic relatedness in P. aeruginosa isolated from patients admitted to a referral hospital in Isfahan, Iran. Materials and Methods: Out of 150 analyzed samples, 54 P. aeruginosa isolates were recovered and were subjected to antibiotic resistance patterns and genetic diversity determination by Kirby-Bauer’s disk diffusion method and RAPD-PCR, respectively. Results: The highest percentage of resistance was observed against ceftazidime and imipenem with 30 (55.6%) isolates; meanwhile all isolates were sensitive to polymyxin B. Twenty-eight (51.8%) isolates revealed resistance to all applied antibiotics. RAPD-PCR (Random Amplified Polymorphic DNA- Polymerase Chain Reaction) results showed 54 unique genotypes, which were divided into 39 clusters. Conclusions: Although different source of P. aeruginosa may involve in patient colonization, genetically related strains were isolated from different wards and or the same ward of the hospital. Our results pointed to the restriction of currently used antibiotics in studied hospital. We hope that our results cast light on the control and transmission of the infection in the investigated hospital.

Background: Pseudomonas aeruginosa is an opportunistic human pathogen, which causes serious problems especially in people who have immunodeficiency. Recently, metallo-β-lactamase (MBLs) resistance in this bacterium has led to some difficulties in treating bacterial infections. The blaSPM-1 is one of the MBL gene families, which induces resistance to the carbapenem class antibiotics; this gene has not been previously assessed in Iran. Objectives: Detection and quantification of blaSPM-1- metallo-β-lactamase gene among resistant Pseudomonas aeruginosa strains (imipenem), isolated from patients in Isfahan hospitals. Patients and Methods: A total of 180 samples were isolated from various nosocomial infections. These isolates were identified as Pseudomonas aeruginosa by using biochemical tests. In order to determine their bacterial drug resistance-pattern the Kirby-Bauer disk diffusion method was utilized. Presence of MBLs in imipenem isolates was detected using the combine disk technique (IMP-EDTA). Similarly, an E-test on Mueller-Hinton agar was used to determine the minimal inhibitory concentration (MIC) of imipenem isolates. The imipenem isolates were then subjected to polymerase chain reaction (PCR) to detect the blaSPM-1 gene. Data were analyzed using the SPSS software (version 16, SPSS Inc., Chicago, IL, USA). Results: In total, 96 isolates of Pseudomonas aeruginosa were collected. Of all isolates, 34 (35.41%) were found to be imipenem-resistant P. aeruginosa. The MIC levels in all imipenem-resistant strains were MIC ≥ 32 μg/mL. Thirteen (38.23%) of the imipenem-resistant P. aeruginosa isolates were MBL positive. None of the isolates carried the blaSPM-1 gene, as indicated by the PCR assay. Conclusions: The rate of imipenem resistance due to MBL has increased dramatically. Early detection and infection-control practices are the best antimicrobial strategy for this organism.

Background: Polycystic Ovary Syndrome (PCOS) is the most common endocrine disorder in 4% - 6% of women in the reproductive age and is a common cause of infertility. Even though the number of investigations is scarce, studies show that Helicobacter pylori infection may influence reproduction. Objectives: The purpose of this study was to determine and compare the levels of H. pylori specific antibodies IgA, IgG and anti-CagA at both PCOS and non-PCOS women with their spouses using the serological test. Patients and Methods: In this cross-sectional study, 127 women with their spouses (age range, 30 - 60 years) were selected. These patient were referred to infertility center of Shariati Hospital in Tehran, Iran, with a diagnostic criteria of PCOS based on Androgen Excess Society (AES). The specific antibodies of IgA, IgG and anti-CagA were measured using the commercial Enzyme-Linked Immunosorbent Assay (ELISA) kit. Results: The positive titers of H. pylori antibodies IgA, IgG and anti-CagA in the PCOS group were 45 (35%), 79 (62%) and 77 (60.5%), respectively, while in non-PCOS group were 38 (30%), 76 (60%) and 50 (39.5%), respectively. The sera positive for IgA, IgG and anti-CagA antibodies in spouses of the non-PCOS group were 38 (30%), 84 (66%) and 79 (62%) respectively, but in spouses of the PCOS group were 51 (40%), 83 (66%) and 48 (38%), respectively. The results showed that H. pylori infection probably did not affect infertility or reproduction. Conclusions: Findings of this study demonstrate no significant difference between levels of H. pylori specific antibodies of IgA, IgG, anti-CagA and the presence of PCOS disorders, and also indicate that serologic testing is a sensitive method for the detection of H. pylori antibodies. The high prevalence of H. pylori positive antibody levels in both PCOS and non-PCOS patients can be probably associated with the high frequency of H. pylori infection.

known to be highly resistant to current antibiotics due to both intrinsic and acquired resistance mechanisms within the bacteria. Objectives: This study aimed to detect and determine drug resistance patterns in different P. aeruginosa strains isolated from clinical specimens in hospitals of Isfahan, Iran. Materials and Methods: In this descriptive cross-sectional study, approximately 255 different bacterial isolates were gathered from samples of blood, sputum, urine, wounds, burns, respiratory systems, catheter, eye and peritoneum from the major hospitals of Isfahan, Iran, in 2012. Biochemical tests revealed that 106 of these isolates were P. aeruginosa. The resistance rate of these bacterial strains to different antibiotics was then assessed by antibiogram (kirby-bauer method). Data were analyzed using SPSS (version 16, SPSS Inc., Chicago, IL, USA). Results: Resistance rates of the isolates to various antibiotics were obtained. It was found that cefepime and cefotaxime had the highest resistance rates (100%). However, the resistance rates were also high for the drugs imipenem (58.5%), meropenem (58.5%), ceftazidime (89.6%), aztreonm (96.2%), ciprofloxacin (77.4%) and gentamicin (66%). Moreover, the lowest resistance rate was observed for amikacin (43.4%). Conclusions: The prevalence of P. aeruginosa strains with multiple-drug resistance was very high amongst the clinical samples in the major hospitals of Isfahan. This is quite a worrisome problem and makes the controlling of the strains more difficult.

Background: The aerobic Actinomycetes are a large group of soil-indwelling bacteria that are distributed in world-wide. These Gram-positive bacteria are most commonly associated with opportunistic infections in both immunocompromised and immunocompetent hosts. Materials and Methods: In this study, three phenotypic and deoxyribonucleic acid (DNA) extraction methods for isolation and identification of Nocardia genus were compared. Samples were taken in five different locations of Isfahan′s suburb from hospitals area, parks, agricultural lands, gardens, arid lands with different soil temperature and pH. Results: In this study, showed that slip-buried-method was better than two other phenotypic methods; 14 out of 70 soil samples (20%) were positive for Nocardia spp. DNA of positive samples were extracted with three techniques and DNA extraction by microwave technique was better than others. This technique was confirmed with observation of DNA bands on 1% agarose gel. Conclusions: These bacteria are important in immune deficient patients such as cancer patients, transplant recipients, tuberculosis; acquired immunodeficiency syndrome etc., Their affluence is unsteady in different zones of the world. In this study, among the three phenotypic methods for the isolation of Nocardia slip-buried method was better than other methods. Among DNA extraction techniques, DNA extraction by microwave method would be selective method for DNA extraction of Nocardia spp. compared with others techniques.

Background Gastroenteritis is a remarkable hygiene problem worldwide. Bacteria and parasites can cause gastroenteritis-associated disorders. Objectives The aims of study were to survey the most common cause of gastroenteritis in patients referred to Imam Khomeini Hospital of Ilam, Iran. Patients and Methods This descriptive-analytical study was performed during 2012 to 2013. After collecting 2376 stool samples, standard biochemical and microbiological tests were performed. Susceptibility was tested by disc diffusion method agreeing with clinical and laboratory standards institute (CLSI) guidelines. The protozoa were detected by sediment wet-mount method. Results Of 2376 patients, 466 (19.6%) were contaminated with pathogenic bacteria or protozoa.The frequency of microorganisms isolated from the patients were 10.3%, 2.5%, 2.5%, 2.1%, 46.4%, 30.9%, 2.5% and 2.5% for enteropathogenic Escherichia coli (EPEC), Shigella dysenteriae, Klebsiella pneumonia, Yersinia enterocolitica, Entamoeba histolytica, Giardia lamblia, E. coli and Candida spp., respectively. Sensitivities to ciprofloxacin in E. coli and S. dysenteriae strains were 100% and 91.66%, respectively. Conclusions The results showed that some patients were probably contaminated with nonbacterial and nonparasitic agents. All the parasitic isolates were resistant to most antibiotics. Therefore determination of microbial isolates and antibiotic susceptibility is necessary before treatment procedures.

Background: Pseudomonas aeruginosa is an opportunistic human pathogen that causes serious problems, especially in people, who have immunodeficiency. In recent times, metallo-β-lactamase (MBLs) resistance in this bacterium has led to some difficulties in treating bacterial infections. The metallo-beta-lactamase family of genes, including blaVIM-1, is being reported with increasing frequency worldwide. The aim of this study is the detection of the metallo-β-lactamase gene blaVIM-1 in imipenem-resistant P. aeruginosa (IRPA) strains isolated from hospitalized patients. Materials and Methods: In this study, 106 P. aeruginosa samples were isolated from various nosocomial infections. The isolates were identified, tested for susceptibility to various antimicrobial agents by the Kirby-Bauer disk diffusion method, and all the imipenem-resistant isolates were screened for the presence of MBLs by using the combined disk (IMP-EDTA). The minimal inhibitory concentration (MIC) of imipenem was determined by E-test on the Mueller-Hinton agar. To detect the blaVIM-1 gene, the isolates were subjected to a polymerase chain reaction (PCR). Results: Of all the P. aeruginosa isolates, 62 (58.5%) were found to be imipenem-resistant P. aeruginosa (MIC ≥32 μg/ml). Twenty-six (42%) of the imipenem-resistant isolates were MBL positive. None of these isolates carried the blaVIM-1 gene using the PCR assay. Conclusion: The results demonstrated the serious therapeutic threat of the MBL-producing P. aeruginosa populations. The rate of imipenem resistance due to MBL was increased dramatically. Early detection and infection-control practices are the best antimicrobial strategies for this organism. None of MBL-producing isolates in this study carry the blaVIM-1 gene; therefore, another gene in the MBL family should be investigated.

Objective To evaluate antimicrobial resistance, invasion index and genetic profile in Listeria monocytogenes isolated from clinical samples. Methods At all, 170 clinical samples were collected from patients with spontaneous abortions hospitalized in Shariati hospital in Tehran during June 2010 to August 2013. Invasion index was determined using HeLa cells. The multiple-locus variable-number tandem-repeats analysis (MLVA) was used for evaluation of genetic relatedness. Results Out of 14 L. monocytogenes isolates, 4 (28.57%), 2 (14.28%), 0 (0%), 5 (35.71%) and 3 (21.42%) were isolated from placental tissue, urine, blood, vaginal and rectal swabs, respectively. High resistance to penicillin and multidrug resistant were found amongst isolates. The invasion index was in the range of 0.001–0.007. Seven different types were obtained by MLVA assay and type 2 and 3 with 4 strains were the most frequent type. Strains isolated from the vagina and the placenta of the same type were also more resistant to penicillin. Conclusions Since MLVA is a high-throughput screening method that is fairly inexpensive, easy to accomplish, rapid, and trustworthy, it is well suited to interlaboratory comparisons during epidemiological investigations. Also further studies of larger samples from a variety of sources such as food and animal specimens recommended comparing by MLVA method.

INTRODUCTION: This study aimed to determine the prevalence, and virulence factors of Listeria monocytogenes isolated from various samples by multiplex polymerase chain reaction (MPCR). METHODS: A total of 617 isolates were obtained and MPCR was employed for detection of the inlA, inlC, and inlJ genes. RESULTS: L. monocytogenes was detected in 46 (7.45%) of the 617 specimens. inlA, inlC, and inlJ were detected in 100%, 76.26%, and 71% isolates, respectively. CONCLUSIONS: This study validated MPCR in the analysis and rapid detection of L. monocytogenes. The role of the genes in pathogenesis of the strains can also be affirmed.