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Background: In recent decades, because of increasing prevalence of immunocompromised disease, non-tuberculous mycobacteria that have been known already as environmental bacteria presented as an important pathogens. In this study, the prevalence of NTM in Isfahan city water samples was determined bydifferent methods including culture methods with biochemical tests. Materials and Methods: Eighty-five water samples were collected from different sources in Isfahan, Iran. The mycobacterial isolates were identified by the growth characteristics, pigment production, semi-quantitative catalase test, Tween 80 hydrolysis, arylsulfatase test (3 and 14 days), heat-stable catalase (pH 7, 68°C), pyrazin amidase (4 and 7 days), urease, nitrate reduction test, and colony morphology. Results: Chlorine concentrations of the samples ranged from 0 to 1.8 mg/l. Twenty-one culturable species including M. fortuitum, 23.8% (5 cases); M.smegmatis, 14.3% (3 cases); M. chelonae like organisms, M. terrae complex, M. gordonae and M. mucogenicum, 9.5% (2 cases for each species); M. avium complex, M.phlei, M. xenopi, M. fallax, and M. flavescenc 4.8% (1 case for each species) were identified. Conclusion: The results of this study showed the incidence of different species of NTM in this geographical region in Iran. Studies show that the prevalence of immunocompromised disease is increasing in the community and it needs different treatment management strategy; therefore, the results of this study can be useful in this strategy.

Pseudomonas aeruginosa has different antibiotic resistance pathways, such as broad-spectrum lactamases and metallo-β-lactamases (MBL), penicillin-binding protein (PBP) alteration, and active efflux pumps. Polymerase chain reaction (PCR) and sequencing methods were applied for double-locus sequence typing (DLST) and New Delhi metallo-β-lactamase (NDM) typing. We deduced the evolutionary pathways for DLST and NDM genes of P. aeruginosa using phylogenetic network. Among the analyzed isolates, 62.50% of the P. aeruginosa isolates were phenotypically carbapenem resistance (CARBR) isolates. Characterization of isolates revealed that the prevalence of bla NDM , bla VIM , bla IMP , undetermined carbapenemase, and MexAB-OprM were 27.5%, 2%, 2.5%, 12.5%, and 15%, respectively. The three largest clusters found were DLST t20-105, DLST t32-39, and DLST t32-52. The network phylogenic tree revealed that DLST t26-46 was a hypothetical ancestor for other DLSTs, and NDM-1 was as a hypothetical ancestor for NDMs. The combination of the NDM and DLST phylogenic trees revealed that DLST t32-39 and DLST tN2-N3 with NDM-4 potentially derived from DLST t26-46 along with NDM-1. Similarly, DLST t5-91 with NDM-5 diversified from DLST tN2-N3 with NDM-4. This is the first study in which DLST and NDM evolutionary routes were performed to investigate the origin of P. aeruginosa isolates. Our study showed that the utilization of medical equipment common to two centers, staff members common to two centers, limitations in treatment options, and prescription of unnecessary high levels of meropenem are the main agents that generate new types of resistant bacteria and spread resistance among hospitals.

Background: Enterococci may develop resistance to linezolid through chromosomal mutations that involve specific linezolid resistance genes, such as cfr , optrA , and poxtA . The objective of this study was to evaluate the antibiotic susceptibility of enterococcal isolates and identify cfr , optrA , and poxtA genes in MDR isolates., Materials and Methods: Enterococcal isolates were collected from various clinical specimens at Al-Zahra, Amin, and Khorshid Hospitals in Isfahan. The Enterococcus isolates were identified as belonging to the E. faecalis and E. faecium species by using specific gene (D alanine D alanine ligase ddl ) sets in PCR. To detect cfr , optrA , and poxtA genes among the species, a multiplex-PCR assay was performed., Results: Out of 175 isolates, E. faecalis predominated 129/175 (73.7%). Furthermore, the prevalence of vancomycin-resistant Enterococci (VRE) and linezolid-resistant Enterococci (LRE) was 29.7% and 4%, respectively. The overall prevalence of MDR was 91.1%, 68.9%, and 66.6% of E. faecium , E. faecalis , and other Enterococcus spp ., respectively. Interestingly, the frequency of optrA (71.4%) in E. faecium and poxtA and crf (42.8%) in E. faecalis were detected among LRE species. A statistically significant relationship ( P < 0.05) was found between the presence of the three genes and the occurrence of LRE., Conclusion: This is the first study to report the detection of linezolid resistance genes ( cfr , optrA , and poxtA ) in clinical Enterococcus spp . isolates from Iran, conducted at Isfahan University of Medical Sciences hospitals. The emergence of enterococcal strains that resist linezolid is concerning as it can lead to the spread of resistant strains among patients, resulting in treatment failure., Competing Interests: There are no conflicts of interest., (Copyright: © 2023 Advanced Biomedical Research.)

Background: This study aimed to compare the antimicrobial effects of green tea, microwaving, cold boiled water, and chlorhexidine (CHX) on Streptococcus mutans and Candida albicans on silicone pacifiers., Materials and Methods: In this in vitro experimental study, 60 equal-size samples of silicone pacifiers were cut, ultraviolet sterilized, and randomly divided into two groups ( n = 30) for immersion in 0.5 McFarland standard suspension of S. mutans and C. albicans . The samples in each group were then randomly divided into five subgroups ( n = 6) for disinfection with 0.12% CHX, cold boiled water, green tea, microwaving for 7 min, and distilled water. The sample suspensions were cultured on blood agar (for S. mutans ) and Sabouraud dextrose agar (for C. albicans ) and incubated. The number of colonies was counted after 24 and 48 h. Data were analyzed using the Kruskal-Wallis and Mann-Whitney tests ( P < 0.05)., Results: At 24 and 48 h, the S. mutans colony count was the lowest in CHX and green tea subgroups followed by microwave, cold boiled water, and distilled water subgroups ( P < 0.05)., Conclusion: CHX and green tea can significantly decrease the S. mutans and C. albicans colony count on silicone pacifiers., Competing Interests: The authors of this manuscript declare that they have no conflicts of interest, real or perceived, financial or nonfinancial in this article., (Copyright: © 2022 Dental Research Journal.)

Introduction: Early childhood caries is a kind of caries occurring in deciduous teeth. Bacteria are among the main factors. Antibacterial agents such as fluoride are used in both prevention and treatment, but their application in children faces limitations such as fluorosis. Therefore, novel methods of caries prevention among the children are mainly focused on the use of fluoride-free active ingredients. In this comparative study, antibacterial effects of gels containing propolis and aloe vera, fluoride, xylitol, and CPP-ACP were investigated., Methods: This is an in vitro study. By plate well technique, plates containing gels were created in the culture medium of Streptococcus mutans and Lactobacillus , and their antibacterial impacts were evaluated by measuring the inhibition zone after 24, 48, and 72 hours. Then, different concentrations of each gel were evaluated in the same way for the antibacterial properties. For each sample, this process was iterated 3 times, where the average was declared as the final number. The collected data were entered in SPSS 24., Results: In both bacteria, propolis gel and aloe vera had the highest zone of inhibition, followed by fluoride and xylitol in the second and third places, respectively. Different concentrations of gels are significantly different in terms of antimicrobial effect ( P value ≤ 0/05). The antimicrobial effect of propolis and aloe vera gel was kept up to the concentration of 1/16. As the bacterial and gel contact time is prolonged, the antibacterial effect of different gels increases, but the difference is not statistically significant ( P value = 0.109). CPP-ACP gel had no antimicrobial effect at any concentration., Conclusion: Propolis and aloe vera gel had a greater antimicrobial effect than other gels, where such effect was observed in low concentrations. CPP-ACP gel had no antimicrobial properties., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2021 Maryam Hajiahmadi et al.)

New Delhi metallo-β-lactamase variants and different types of metallo-β-lactamases have attracted enormous consideration for hydrolyzing almost all β-lactam antibiotics, which leads to multi drug resistance bacteria. Metallo-β-lactamases genes have disseminated in hospitals and all parts of the world and became a public health concern. There is no inhibitor for New Delhi metallo-β-lactamase-1 and other metallo-β-lactamases classes, so metallo-β-lactamases inhibitor drugs became an urgent need. In this study, multi-steps virtual screening was done over the NPASS database with 35,032 natural compounds. At first Captopril was extracted from 4EXS PDB code and use as a template for the first structural screening and 500 compounds obtained as hit compounds by molecular docking. Then the best ligand, i.e. NPC120633 was used as templet and 800 similar compounds were obtained. As a final point, ten compounds i.e. NPC171932, NPC100251, NPC18185, NPC98583, NPC112380, NPC471403, NPC471404, NPC472454, NPC473010 and NPC300657 had proper docking scores, and a 50 ns molecular dynamics simulation was performed for calculation binding free energy of each compound with New Delhi metallo-β-lactamase. Protein sequence alignment, 3D conformational alignment, pharmacophore modeling on all New Delhi metallo-β-lactamase variants and all types of metallo-β-lactamases were done. Quantum chemical perspective based on the fragment molecular orbital (FMO) method was performed to discover conserved and crucial residues in the catalytic activity of metallo-β-lactamases. These residues had similar 3D coordinates of spatial location in the 3D conformational alignment. So it is posibble that all types of metallo-β-lactamases can inhibit by these ten compounds. Therefore, these compounds were proper to mostly inhibit all metallo-β-lactamases in experimental studies.

This study investigated carbapenem resistance among Acinetobacter baumannii isolated from respiratory specimens. Epidemiological relationship of the isolates was also evaluated. In this study, 81 respiratory specimens of A. baumannii from AL Zahra Hospital were confirmed by phenotypic and genotypic methods. Antimicrobial susceptibility was performed by disc diffusion method. Carbapenem resistance genes were identified by PCR. The isolates were typed by RAPD-PCR and multilocus sequence typing (MLST) methods. All isolates were resistant to imipenem and 80 isolates to meropenem. Frequency of oxacillinase genes was as follows: blaOXA-23 gene was positive in 74 (91.3%), blaOXA-24 gene in 50 (61.7%) and blaOXA-58 was not found in any isolates. On the other hand 22 (27.2%) isolates contained blaIMP-1, 3 (3.7%) isolates contained blaIMP-2 gene, 5 (6.2%) isolates contained blaVIM-1, 4 (5%) isolates had blaVIM-2 and none of the isolates had blaSIM-1 gene. RAPD-PCR typing identified 16 different patterns, with one pattern being the most frequent one in 26 isolates. In MLST 6 different sequence types were identified, the most predominant being ST2 belonging to clonal complex 2. The results of this study showed high resistance to carbapenems as well as high abundance of oxacillinase genes.

Integrons are mobilizable platforms-DNA elements with impacts on moving antibiotic resistance genes among bacteria and capable of spreading multi-drug resistance (MDR) in pathogens. Methicillin-resistant Staphylococcus aureus (MRSA) strains are the main cause of community-acquired and nosocomial infections with high mortality and morbidity rates worldwide. This work is mainly aimed at calculating the frequency of Type I, II, and III integrons within multi-drug resistance and Methicillin-resistant S. aureus Isolates in Iran. In this cross-sectional study, 230 clinical isolates of S. aureus were gathered from patients of educational hospitals in the provinces of Iran. These isolates were verified utilizing particular biochemical examinations and then assessed for antibiotic susceptibility through disk diffusion technique and standard procedures were done. Genomic and plasmid DNA of all isolates were extracted using Extraction Kit and PCR assay was used for the detection of Type I, II and III integrons genes. Out of the 230 S. aureus isolates, 136 (59.1%) isolates were MRSA and 141 (61.3%) isolates exhibited the MDR pattern. PCR and sequencing showed that 57 (24.8%) of tested isolates carry Type I integron. Among the isolates investigated, MRSA and MDR isolates showed frequencies of 56.1% and 57.9%, respectively. Type II and III integrons were found in none of 230 isolates. The IntI I gene was present in approximately one-quarter of this study isolates. The great prevalence rate of MDR and MRSA isolates and concurrently the existence of Type I integron among those isolates have been considered an important concern in medical society.

Background: Fluoroquinolone resistant Escherichia coli isolates have become an important challenge in healthcare settings in Iran. In this study, we have determined Fluoroquinolone resistant E. coli isolates (from both outpatients and inpatients) and evaluated mutations of gyrA and parC within the quinolone resistance-determining regions (QRDR) of these clinical isolates., Materials and Methods: Clinical isolates were recovered from the urine sample of patients with urinary tract infections admitted at Alzahra hospital, Iran, between September and February 2013. We assessed antimicrobial susceptibility of all isolates and determined mutations in QRDR of gyrA and parC genes from 13 fluoroquinolone-resistant isolates by DNA sequencing., Results: A total of 135 E. coli strains were obtained from 135 patients (91 outpatients and 44 inpatients). The resistance rate of fluoroquinolones (Ciprofloxacin, Norfloxacin and Ofloxacin) among our strains was 45.2%. Two E. coli isolates were shown just a single mutation, but other isolates possessed 2-5 mutations in gyrA and parC genes. Mutations in the QRDR regions of gyrA were at positions Ser83 and Asp87 and parC at positions Ser80, Glu84, Gly78., Conclusions: Ciprofloxacin is the most common antimicrobial agent used for treating urinary tract infections (UTIs) in healthcare settings in Iran. Accumulation of different substitutions in the QRDR regions of gyrA and parC confers high-level resistance of fluoroquinolones in clinical isolates.

Following publication of the original article [1], an error was reported in the tables due to a typesetting mistake. Table 3 is presented as a duplicate of Table 2. In this Correction, the correct presentation of Table 3 is shown.

The objective of this study was to assess the prevalence, antibiogram, and related genes of carbapenem-resistant Klebsiella pneumoniae (CRKP) among hospital environment samples. A total of 250 samples were taken from different surfaces and medical devices of three hospitals in Isfahan, Iran. All samples were cultured and K. pneumoniae strains were identified by conventional microbiological methods and polymerase chain reaction (PCR). Antibiogram of isolates was performed by disk diffusion method and production of carbapenemases and metallo-β-lactamases (MBLs) was confirmed using modified Hodge test and E-test, respectively. Molecular detection of the related genes was carried out by PCR. Overall, 37 (14.8%) K. pneumoniae strains were isolated, of which 34 (91.9%) strains were resistant to carbapenems. Twenty-eight (82.4%) isolates were positive for carbapenemases and seven (20.6%) isolates were phenotypically MBL producers. The results of PCR showed that the prevalence of bla OXA-48 , bla NDM , bla IMP , bla SHV , bla CTX-M , bla TEM , and class 1 integron among CRKP isolates was 70.6%, 52.9%, 2.9%, 100%, 82.4%, 55.9%, and 76.5%, respectively. However, bla KPC , bla GES , bla IMI , bla VIM , and class 2 integron were not detected in any of the isolates. This study showed that the environment of our hospitals is contaminated with CRKP and it emphasizes the importance of using standard methods for infection control.

Objectives: The role of integrons in the transfer of antibiotic resistance is one of the important issues, therefore, this study is aimed to investigate antibiotic resistance pattern and prevalence of class 1 and 2 integrons in P. aeruginosa isolated., Results: Out of 72 confirmed P. aeruginosa isolates, 50% were from ICU patients. Antibacterial susceptibility pattern showed that isolates were most resistant to ceftazidime (76.4%) and colistin was the most effective antibiotic (100%) and molecular analysis of class I and II integrons showed 55.5% and 29.1% of isolates were positive, respectively and the proportions of MDR isolates were significantly higher among integron-positive isolates with 73.6% compared to negative isolates with 22.9%. Our results showed that there was a correlation among class 1 and 2 integrons with MDR P. aeruginosa isolates. According to the importance of integrons in acquisition and dissemination of antibiotics resistance genes, the performance of antibiotic surveillance programs and investigating the role of integrons is recommended to control the spreading of antibiotics resistance genes.

Nocardiae are Gram-positive, filamentous, aerobic, relatively slow-growing, and weakly acid-fast bacteria which cause nocardiosis in humans. We describe a 53-year-old patient with chronic bronchitis referred to Al-Zahra Hospital, Isfahan. A bronchial washing sample was taken from the patient. A Nocardia -like microorganism was detected in microscopic evaluation. Based on the phenotypic and 16S rRNA gene sequencing, the isolate was identified as Nocardia thailandica . The patient was treated with trimethoprim-sulfamethoxazole and linezolid. This is the first report of the isolation of Nocardia thailandica from Iran.

The aim of this study was to investigate the prevalence of carbapenem-resistant Klebsiella pneumoniae (CRKP) and the most common types of carbapenemases, metallo-beta-lactamases (MBLs), and extended-spectrum beta-lactamases (ESBLs) among CRKP isolates in a tertiary hospital in Isfahan, Iran. Eighty non-repetitive clinical isolates of K. pneumoniae were obtained from different clinical specimens. Antibiotic resistance pattern of isolates was determined by disk diffusion method and production of carbapenemases and MBLs was confirmed using modified Hodge test and E-test, respectively. Molecular detection of the antibiotic resistance genes was performed using PCR. Fifty-one (63.8%) isolates have decreased susceptibility to carbapenems, of which 46 (90.2%) isolates were as carbapenemase producer and four (7.8%) isolates were positive for MBLs, phenotypically. The results of PCR showed that the prevalence of bla OXA-48 , bla NDM , bla SHV , bla CTX-M , and bla TEM genes among CRKP isolates were 90.2%, 15.7%, 98%, 96.1%, and 90.2%, respectively. No isolates carrying the bla KPC , bla GES , bla IMI , bla VIM , and bla IMP genes were detected. This study showed that the production of OXA-48 is one of the main mechanisms of resistance to carbapenems in CRKP isolates in Isfahan. In addition, the dissemination of NDM-producing CRKP isolates is a potential risk for the health care system of this area in the near future.

Introduction: The emergence of Metallo-beta-lactamase (MBL)-producing Acinetobacter baumannii has become a global concern in nosocomial infections. The aim of this study is to determine the prevalence of MBL producing genes among clinical isolates of A. baumannii from hospitalized patients., Methods: This study was performed from October 2015 to October 2016 at three teaching hospitals located in Isfahan, Iran. Totally, 100 A-baumannii isolates were collected from clinical specimens and identified as A-baumannii using standard microbiological methods. Antimicrobial susceptibility test was determined by disc diffusion method according to the CLSI. Furthermore, the determination of bla IMP-1 , bla IMP-2 , bla VIM-1 , bla VIM-2 and bla SIM-1 was detected by PCR., Results: Totally, Sixty-eight percent (68%) of isolates of A. baumannii were recovered from tracheal aspirate. According to the antibiotic susceptibility pattern, the highest level of resistance was against ciprofloxacin (99%), while among tested antibiotics amikacin (10%) was found to be the most effective. 21%, 4%, 7% and 6% isolates carried bla IMP-1 , bla IMP-2 , bla VIM-1 and bla VIM-2 genes, respectively. Also, bla SIM-1 was not detected in any of the isolates., Conclusion: The results of this study showed high rate of the MBL producing A-baumannii isolates in our region and displayed that MBLs producing A-baumannii strains are emerging threats to ICUs.

Acinetobacter baumannii is one of the most important bacterial species with the ability to produce OXA-type carbapenemases. We aimed to evaluate the prevalence of OXA-type carbapenemases among clinical isolates of A. baumannii in three major hospitals of Isfahan. In this cross-sectional descriptive study, 153 non-repeated strains of A. baumannii were isolated from various clinical samples of hospitalized patients in Al-Zahra, Imam Mousa Kazem, and Shariati hospitals from October 2015 to October 2016. Antimicrobial susceptibility testing for imipenem, meropenem, ertapenem, cefepime, ceftazidime, ceftriaxone, piperacillin-tazobactam, gentamicin, amikacin, ciprofloxacin, and tetracycline was performed using the disk diffusion method. In order to identify bla-oxa genes, a multiplex polymerase chain reaction was used. The resistance rates in A. baumannii isolates to beta-lactam antibiotics including imipenem, ertapenem, meropenem, cefepime, ceftazidime, ceftriaxone, and piperacillin/tazobactam were 100%, 100%, 99.3%, 97.4%, 96.7%, 97.4%, and 98.6%, respectively. PCR assay showed the presence of bla-oxa genes in all isolates. The bla-oxa-51 gene was recognized in all (100%) isolates, 90.8% and 62.1% of isolates possessed the bla-oxa-23 and bla-oxa-24 genes, respectively, while the bla-oxa-58 gene was not detected in any of the isolates. Also, 56.2% of isolates had both the bla-oxa-23 and bla-oxa-24 genes simultaneously. We found that the prevalence of OXA-type carbapenemases among carbapenem-resistant A. baumannii isolates is high in Isfahan, with OXA-23 being the major carbapenemase mechanism responsible for the resistance phenotype.

Background: Extended-spectrum β-lactamase (ESBL)-producing is a significant resistant mechanism to β-lactams in Enterobacteriaceae , especially in Klebsiella pneumoniae . The main objectives of this study were to genetically characterize urinary clinical isolates of K. pneumoniae through the investigating of blaTEM , blaCTX-M and using molecular typing by Enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) method. We also determined the frequency of antibiotic resistance of K. pneumoniae strains to characterize the β-lactamases included., Materials and Methods: A cross-sectional study was carried out to evaluate 98 strains of K. pneumoniae isolated from urine culture of outpatients referred to Al-Zahra Hospital, Isfahan, Iran. Antibiotic susceptibility testing was performed using Kirby-Bauer's method. Screening of ESBLs was carried out using double-disk screening test. PCR technique was performed to detect TEM and CTX-M genes. The total DNA of each strain was tested by ERIC-PCR., Results: In 98 K. pneumoniae studied clinical isolates, 25.5% were ESBL producing and 44.9% multidrug-resistant (MDR). From 25 ESBL isolates, 23 (92%) cases showed MDR phenotype. In ESBL producing isolates, 23 (92%) were blaCTX-M and 19 (76%) blaTEM positive. The antimicrobial drug susceptibilities of ESBL isolates indicated high resistant rates for cefotaxime and ceftazidime. All 25 ESBL producing isolates were resistant to cefotaxime. Complex patterns of fingerprints isolates showed that 36% of the isolates were belonged to the cluster no 5., Conclusion: This study revealed high antimicrobial resistance rates among ESBL isolates which can lead to various health difficulties. Epidemiological data collection from patients is recommended to develop the strategies to manage antibiotic resistance., Competing Interests: There are no conflicts of interest.

Objectives: This study aimed to determine the prevalence of class I, II, and III integrons among clinical Acinetobacter baumannii isolates collected from hospitalized patients., Methods: This cross-sectional study was conducted at two teaching hospitals in Isfahan, Iran, from October 2015 to October 2016. A total of 147 non-duplicate A. baumannii isolates were collected from clinical specimens and identified as A. baumannii using standard microbiological methods and confirmed by genotyping. Antimicrobial susceptibility was determined using disc diffusion method, and the presence of integron genes was performed using the polymerase chain reaction., Results: Out of 147 confirmed A. baumannii isolates, 97.3% of isolates were extensive drug-resistant (XDR) and 2.7% were multidrug-resistant (MDR). Class I and II integrons were detected in 63.9% and 78.2% of the A. baumannii , respectively. Class III integron was not detected in any of the isolates., Conclusion: Our results show a high prevalence of classes I and II integrons which may play a key role in the acquisition of MDR and XDR phenotype among A. baumannii isolates in our region. Therefore, use of appropriate infection control in clinical settings and implementation of treatment strategies is necessary for our hospitals.

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.

Background and Objectives: Urinary tract infection (UTI) is one of the most frequent infectious diseases and can occur in all age groups. Escherichia coli is the main cause of this infection. Multiple resistances to antimicrobial agents are increasing quickly in E. coli isolates and may complicate therapeutic strategies for UTI. The aim of this study was to determine the antibiotic resistance pattern and the multidrug-resistance (MDR) phenotypes in uropathogenic E. coli (UPEC)., Materials and Methods: A total of 135 UPEC isolates were collected from both outpatients (91 isolates) and inpatients (44 isolates) between September, 2012 and February, 2013. In order to determine the MDR among UPEC isolates, we have tested 15 antimicrobial agents and antibiotic susceptibility was done by Kirby-Bauer disk diffusion method., Results: The percentage of MDR isolates (resistant to at least three drug classes such as aminoglycosides, fluoroquinolones, penicillins, cephalosporins, or carbapenems) was 68% in the inpatients and 61% in the outpatients. Antibiotic resistance to ampicillin, ceftazidim, nalidixic acid, and trimethoprim/sulfamethoxazole were higher than 50%. Amikacin, nitrofurantoin, and gentamicin showed markedly greater activity (89.1%, 85.9%, and 82.4% sensitivity, respectively) than other antimicrobial agents. Resistance to meropenem did show either in outpatients or in inpatients., Interpretation and Conclusions: The high prevalence of drug resistance among UTI patients calls for continuous monitoring of the incidence of drug resistance for appropriate empiric selection of antibiotic therapy. Empirical treatment of UTIs should be relied on susceptibility patterns from local studies.

Background: Helicobacter pylori infection is highly prevalent in the developing countries. It causes gastritis, peptic ulcer disease, and gastrocarcinoma. Treatment with drugs and antibiotics is problematic due to the following reasons: cost, resistance to antibiotics, prolonged treatment and using multiple drugs. Catalase is highly conserved among the Helicobacter species and is important to the survival of the organism. It is expressed in high amounts and is exposed to the surface of this bacterium; therefore it represents a suitable candidate vaccine antigen., Objectives: A suitable approach in H. pylori vaccinology is the administration of epitope based vaccines. Therefore the responses of T-cells (IFN-γ and IL-4 production) against the catalase of H. pylori were determined. Then the quality of the immune responses against intact catalase and three epitopes of catalase were compared., Materials and Methods: In this study, a composition of three epitopes of the H. pylori catalase was selected based on Propred software. The effect of catalase epitopes on T-cells were assayed and immune responses identified., Results: The results of IFN-γ, IL-4 production against antigens, epitopes, and recombinant catalase by T-cells were compared for better understanding of epitope efficiency., Conclusions: The current research demonstrated that epitope sequence stimulates cellular immune responses effectively. In addition, increased safety and potency as well as a reduction in time and cost were advantages of this method. Authors are going to use this sequence as a suitable vaccine candidate for further research on animal models and humans in future.

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 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.

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: Multidrug resistance Pseudomonas aeruginosa (MDR-P. aeruginosa) is a worldwide threat for public health. Hyperexpression of efflux pump systems (MexAB-OprM and MexCD-OprJ), which is a well-known mechanisms for MDR emerging, is controlled by regulatory genes, mexR and nfxB, respectively. The aim of this study was to evaluate point mutations in mexR and nfxB genes in MDR- P. aeruginosa isolated from wound infections., Materials and Methods: A total of 34 P. aeruginosa cultures obtained from wound infections were analyzed. Among them eight isolates identified as MDR-P. aeruginosa and were subjected to determination of mutations in mexR and nfxB genes., Results: We detected eight-point mutations in mexR and 12-point mutations in nfxB. The most common mutations were common G327-A (eight isolates), G384-A (eight isolates), G411-A (eight isolates). Mutations in A371-C and A372-C were the predominant substitution which was seen in nfxB. Amino acid substitutions were also found at position 124 and 126 for NfxB and MexR, respectively., Conclusions: P. aeruginosa isolates with mutation in efflux pump regulatory genes such as mexR and nfxB could be a main factor contributed to antibiotic resistance and must be considered in antibiotic treatment.

Background: Helicobacter pylori antimicrobial resistance is an important factor responsible for treatment failure. The purpose of this study was evaluating the prevalence of point mutations in clarithromycin-resistant clinical isolates of H. pylori in Isfahan city of Iran., Materials and Methods: Thirty isolates of H. pylori from 130 biopsy specimens were isolated by culture and confirmed by biochemical and PCR tests. The MIC of clarithromycin antibiotic for 30 clinical isolates of H. pylori was determined by E-test method. The point mutations in the 288 bp of 23S rRNA gene of H. pylori were investigated in four clarithromycin-resistant clinical isolates by PCR followed by sequencing., Results: Among 30 isolates of H. pylori, 4 cases were resistant to clarithromycin. One point mutation was found at position T2243C in the 23S rRNA gene in all resistance isolates., Conclusions: In our study, H. pylori resistance to clarithromycin associated with point mutation at position 2243 (T2243C).

Background: Helicobacter pylori (H. pylori) is a spiral Gram negative bacteria that can transform to the coccoid form in adverse conditions., Objectives: The aim of this study was to determine the in vitro morphological and bactericidal effects of metronidazole, amoxicillin and clarithromycin on H. pylori., Materials and Methods: The standard strain 26695 of H. pylori was cultured on Brucella agar (BA) and the minimum inhibitory concentrations (MICs) of three antibiotics were determined by E-test method. The bacteria were exposed to antibiotics at 1/2 MIC, MIC and 2X MIC concentrations in Brucella broth (BB). Induced coccoid forms were confirmed by Gram staining and light microscopy. The viability of cells as well as the susceptibility of viable coccoids to antibiotics were examined using the flow cytometry method., Results: All of the three antibiotics at sub-MIC induced coccoid forms. The highest rates of coccoids (> 90%) were induced at 0.008 μg/mL concentration (1/2 MIC) of amoxicillin, 72 hours postexposure. Metronidazole and clarithromycin with 1/2 MIC (0.5 and 0.125 µg/mL respectively) induced lower rates of coccoid forms (60% and 40% respectively). Potent bactericidal effects on coccoids were observed with Metronidazole at 2X MIC and clarithromycin at MIC (0.25 µg/mL) (80 - 90%). Amoxicillin with MIC and 2X MIC had no bactericidal effect on coccoid forms., Conclusions: Despite the good in vitro bactericidal effect of amoxicillin on spiral forms of H. pylori, this antibiotic has little effect on induced coccoids that may develop after the inappropriate in vivo antibacterial treatment. Hence, for successful therapy, it is essential not only to eradicate the spiral forms, but to eliminate the viable coccoids.

Helicobacter pylori (H. Pylori) is an actively dividing spiral bacterium that changes to coccoid morphology under stressful environments. The infectivity of the coccoids is still controversial. The aim of this study was to determine the viability and expression of two important virulence genes (babA and cagE), in antibiotic-induced coccoid forms. Three strains of H. pylori, the standard 26695 and two clinical isolates (p1, p2) were converted to coccoid form by amoxicillin. Coccoids were identified according to Gram-staining and microscopic morphology. The viability of the cells was analyzed by flow cytometry. The expression of cagE and babA in coccoid forms were evaluated and compared to the spirals by quantitative PCR assay. The coccoid forms were developed after 72 h exposure of H. pylori to ½ MIC of amoxicillin, and the conversion form was completed (100 %) at 144 h in all of three isolates. Flow cytometry analyses showed that the majority of the induced coccoids (90-99.9 %) were viable. Expression of cagE and babA was seen in coccoids; however, in lower rate (cagE, ~3-fold and babA, ~10-fold) than these in spiral forms. Coccoid forms of two clinical isolates significantly expressed higher rate of cagE and babA than standard 26695 strain (P = 0.01). These results suggest that the induced coccoid form of H. pylori is not a passive entity but can actively infect the human by expression of the virulence genes for long time in stomach and probably play a role in chronic and severe disease.

Background: Throughout the world, bloodstream infections (BSIs) are associated with high rates of morbidity and mortality. Rapid pathogens identification is central significance for the outcome of the patient than culture techniques for microbial identification. To develop an end point multiplex PCR to identify a group of bacteria including Enterococcus spp., Pseudomons aeruginosa, Staphylococcus spp., Acinetobacter baumannii, 16S rDNA, and Drosophila Melanogaster were used as internal control (IC)., Materials and Methods: Design of primers was done using Mega4, Allel ID6, Oligo6 and Oligo analyzer softwares. Genetic targets for primer designing and identification of genus Enterococcus spp., Staphylococcus spp., and species of Acinetobacter baumannii, Pseudomons aeruginosa, included the rpoB, rpoB and gyrA, sss respectively. Then PCR and multiplex PCR were performed., Results: The intended specificity was obtained for the bacteria, which used in this study and there wasn't seen any unspecific amplification by the multiplex PCR. The test showed a sensitivity ranging from 1 to 100 target copies per reaction depending on the bacterial species., Conclusions: The presented multiplex PCR offers a rapid and accurate molecular diagnostic tool for simultaneous detection of some pathogenic microorganisms. The IC exists in the multiplex PCR accompanied by other primers in the system, can serve as a simple, cost- effective internal control for the multiplex PCR assay.

Introduction: Many studies have shown epidemiological links between strains isolated in tap water, and those isolated from patients. Molecular methods linked to PCR are more reliable and faster for identification of non- tuberculous mycobacteria (NTM). In this study molecular methods were used for identification and typing of NTM., Materials and Methods: Five hundred ml of 85 water samples was passed through 0.45 μm filters. The filters were transferred directly onto 7H10 Middle Brook solid media, containing 15% OADC. PCR for 16S rRNA was done and the PCR product (1500 bp) was sequenced. PRA of the hsp65 gene was investigated to identify the species of isolates. For evaluation of susceptibility of NTM to antimycobacterial agents, E-test method was used., Result: The genus of 26 isolated NTM was confirmed by 16s rRNA sequence based method. Nineteen isolates of Mycobacteria were differentiated using hsp65 genes PRA. The dominant isolates were M. fortuitum (26.7%), M. chelonae like organism (13.3%) and M. mucogenicum (13.3%). Seventy one percent of NTM species were resistant to isoniazid, 64% to rifampin, 57% to ethambutol, 35% to tetracycline, 14 % to azithromycin and 7.1 % to amikacin., Conclusion: The results showed that E-test method is not a proper technique for antimycobacterial assay because some NTM species are slow in growing and have no growth on Muller Hinton agar. Regarding the 16S rRNA sequence analysis, the identification of isolates was restricted to the genus level, because 99% similarity within 16S rRNA of two isolates may or may not determine the same species.

Objective: Sepsis is a systemic inflammatory response associated with high mortality rates in the clinical setting. A multiplex endpoint polymerase chain reaction (PCR) based assay for rapid detection of enterobacteriaceae involved in septicemia, which included Internal Control (IC) and 16S rDNA, is presented here. To develop a panel of primers for DNA fragments of 16S rDNA, enterobacteriaceae, IC, and evaluate analytical sensitivity and specificity of the test., Materials and Methods: Primers for amplification of enterobacteriaceae, IC, and16S rDNA were designed, and then PCR was performed. Minimal analytical sensitivity was determined by cloning and colony PCR, and specificity was tested on the basis of their respective standard strains. This study is a cross-sectional Model., Results: Our results showed the rpoB gene as the most promising target for detection of enterobacteriaceae by PCR amplification. Specificity and sensitivity of endpoint PCR were 100%, 100%, and 100%, and 10, 1, and 100 copies/reaction for enterobacteriaceae, IC, and 16S rDNA, respectively., Conclusion: The molecular panel presented offers the advantage of an easy, reliable, and cost-effective system when compared to other molecular detection methods. However, further evaluation is needed. Our assay holds promising for more rapid pathogens related in clinical sepsis.

Objectives: Staphylococcus aureus is a foremost source of numerous nosocomial and community acquired infections. Antibiotic therapy for vancomycin resistant S. aureus (VRSA) can not promise the eradication of infections. Since adhesion is the major route of infections, adhesin based vaccine could suppress S. aureus infections. Fibronectin binding protein A (FnBPA) and clumping factor A (ClfA) are major responsible adhesions involved in S. aureus infections, so they could be candidate vaccine molecules against an extensive range of infections. This project intended to express a new fusion protein construct and analysis of biological activity regarding binding activity., Materials and Methods: pfnbA- ClfA construct was transformed to Escherichia coli BL21 (DE3). Transformant E. coli were grown in LB broth and induced with IPTG and cellular extracts were separated on SDS-PAGE. RT-PCR was performed to verify expression. Binding activity of fusion protein was studied using human gingival fibroblast (HGF) cell line. D1-D3 protein from unpublished study was used as control., Results: The expected fusion protein fragment showed by SDS-PAGE. RT-PCR verified the existence of mRNA relating to expressed fusion protein. Binding activity of S. aureus decreased after treatment of HGF cells with fusion protein., Conclusion: In total, binding activity of fusion protein was approximately two fold lesser than D1-D3 protein. It is supposed that the fusion protein could not be attached to its ligand easily and would be more accessible to antigen presenting cells and consequently protective antibodies will be produced. This project is pending for in vivo infection study in animal model.

Objectives: Disposable soft contact lenses that are commonly used after laser refractive surgery are known to be colonized by bacteria and play a key role in Bacterial Keraitis (BK) pathogenesis. Coagulase-negative staphylococci (CoNS) have been found to be the most common pathogen involved in this postoperative infection. In this study a rapid and a simple assay was developed for studying attachment and accumulation of CoNS on soft contact lenses in vitro using [3H] thymidine., Methods: Thirty-five isolates of CoNS were obtained from 27 laser refractive surgery patients. Twenty-five of these thirty-five CoNS were isolated in multiple cultures. Ten CoNS were isolated in cultures from patients who underwent reoperation. The assay was optimized using a biofilm-producing strain, S. epidermidis RP62A, which was subcultured overnight at 37 degrees C on blood agar medium. Quantitative determination of biofilm production was tested. Presence of the genes icaADB and icaD was determined in all isolates. All isolates were biochemically analyzed using the Phene Plate (PhP) system modified for typing of CoNS. The CoNS isolates were further characterized to species level using ID32Staph.Mann-Whitney rank sum test and chi-square test were used to identify statistical differences in adherence, index, antibiotic susceptibility patterns, and biofilm production or presence of the ica operon between clinically significant isolates and non-postoperative BK isolates., Results: No differences in attachment and accumulation were found between isolates causing BK after laser refractive surgery and contaminant isolates. In addition, there were no differences in the distribution of the ica operon between the two groups, as determined by polymerase chain reaction. Nevertheless, the ability to produce biofilm was found to be present significantly more frequently among BK isolates than among non-postoperative BK isolates., Conclusions: This study shows that the method using radioactive thymidine to analyze adherence of CoNS to soft contact lenses enables detection of differences in the adherence patterns of individual isolates.

Background: Coagulase-negative staphylococci and diphtheroids are normal inhabitants of the outer surface of the human eye. These microorganisms serve as part of the defense mechanism of the ocular anatomy in preventing colonization and infection by pathogenic bacteria. Nevertheless, infections associated with contaminated solutions and cases became serious problems for people who wear soft contact lenses. The aim of this study is to isolate and identify aerobic bacteria, particularly, gram-negative species associated with the use of extended-wear soft contact lenses., Methods: Extended-wear contact lenses were collected, using aseptic technique, from the eyes of individuals after 30 days of extended wear (5-7 day intermittent periods) and were examined for adhered aerobic bacteria., Result: Coagulase-negative staphylococci were isolated from 74% of the lenses. Serratia marcescens was found at an incidence of 10% and Pseudomonas aeruginosa at an incidence of 6%., Conclusions: The presence of species of bacteria, including P. aeruginosa and S. marcescens, which have been associated with daily wear soft contact lenses, solutions, and cases also seem to be associated with extended-wear lenses.