Jundishapur Journal of Microbiology
Volume:8 Issue: 4, Apr 2015

Aminoglycosides are a group of antibiotics that have been widely used in the treatment of life-threatening infections of Gram-negative bacteria.. This study aimed to evaluate the frequency of aminoglycoside resistance genes in Enterococcus and Salmonella strains isolated from clinical samples by PCR.. In this study, 140 and 79 isolates of Enterococcus and Salmonella were collected, respectively. After phenotypic biochemical confirmation, 117 and 77 isolates were identified as Enterococcus and Salmonella, respectively. After the biochemical identification of the isolates, antibiotic susceptibility for screening of resistance was done using the Kirby-Bauer method for gentamicin, amikacin, kanamycin, tobramycin and netilmycin. DNA was extracted from resistant strains and the presence of acc (3)-Ia, aac (3′)-Ib, acc (6)-IIa, 16SrRNA methylase genes (armA and rat) was detected by PCR amplification using special primers and positive controls.. Enterococcus isolates have the highest prevalence of resistance to both kanamycin and amikacin (68.4%), and Salmonella isolates have the highest prevalence of resistance against kanamycin (6.9%). Ninety-three and 26 isolates of Enterococcus and Salmonella at least were resistant against one of the aminoglycosides, respectively. Moreover, 72.04%, 66.7%, and 36.6% of the resistant strains of Enterococcus had the aac (3′)-Ia, aac (3′)-IIa, and acc (6′)-Ib genes, respectively. None of the Salmonella isolates have the studied aminoglycoside genes.. Our results indicate that acetylation genes have an important role in aminoglycoside resistance of the Enterococcus isolates from clinical samples. Moreover, Salmonella strains indicate very low level of aminoglycoside resistance, and aminoglycoside resistance genes were not found in Salmonella isolates. These results indicate that other resistance mechanisms, including efflux pumps have an important role in aminoglycoside resistance of Salmonella.

Enterococcal species have emerged as important pathogens in Iran as well as throughout the world. With the increased use of vancomycin, Vancomycin-Resistant Enterococci (VRE) has become an important nosocomial pathogen.. The aim of the present study was to determine the incidence and antimicrobial susceptibility pattern of VRE and also to determine the most important genes that cause resistance to vancomycin in clinical samples in Arak, Iran.. In total, 200 enterococci samples were collected from clinical specimens of Arak hospitals. Enterococcal species were identified using standard biochemical tests. Antibiotic susceptibility was tested by the Clinical and Laboratory Standards Institute (CLSI) disk diffusion. Minimum Inhibitory Concentration (MICs) was determined by broth micro dilution. All of the VRE isolates were examined by PCR to detect the presence of VRE genes.. Disk diffusion agar showed that 96 strains (48%) were resistant to gentamicin, 89 (44.5%) to ciprofloxacin, 127 (63.5%) to erythromycin, 142 (71%) to tetracycline, 11 (5.5%) to teicoplanin, 32 (16%) to vancomycin, none to linezolid and 96 (48%) to co-trimoxazole. The MICs of the resistant isolates were as follows; 88 strains had MIC ≥ 32 μg/mL to vancomycin and 59 strains had MIC ≥ 32 μg/mL to teicoplanin. Molecular studies revealed that 59.09% of VRE contained VanA genes and 7.95% of VRE contained the VanB genes. None of the strains had vanC1 and vanC2/3 gene.. According to the results of this study, rates of vancomycin-resistance in enterococci, in Iran like other parts of the world, is increasing. Therefore accurate methods are required for identifying strains that possess resistance genes because many cases of hospital infections are caused by these strains..

Ophthalmic pterygium is a common benign lesion of unknown origin and the pathogenesis might be vision-threatening. This problem is often associated with exposure to solar light. Recent evidence suggests that potentially oncogenic viruses such as human papillomavirus and Epstein-Barr virus may be involved in the pathogenesis of pterygia. Expression of specific adenovirus genes such as E1A and E1B, which potentially have many functions, may contribute to their oncogenic activity as well as relevance to cellular immortalization.. For the first time, we aimed to investigate involvement of adenoviruses in pterygium formation..Patients and Fifty tissue specimens of pterygium from patients undergoing pterygium surgery (as cases), 50 conjunctival swab samples from the same patients and 10 conjunctival biopsy specimens from individuals without pterygium such as patients undergoing cataract surgery (as controls) were analyzed for evidence of adenovirus infection with polymerase chain reaction using specific primers chosen from the moderately conserved region of the hexon gene. Furthermore, β-globin primers were used to access the quality of extracted DNA. Data was analyzed using SPSS (version 16) software.. Of 50 patients, 20 were men and 30 women with mean age of 61.1 ± 16.9 years ranged between 22 and 85 years. All samples of pterygia had positive results for adenoviruses DNA with polymerase chain reaction, but none of the negative control groups displayed adenoviruses. The pterygium group and the control groups were β-globin positive. Direct sequencing of PCR products confirmed Adenovirus infection.. Adenoviruses might act as a possible cause of pterygium formation and other factors could play a synergistic role in the development. However, further larger studies are required to confirm this hypothesis..

Antibiotic resistance is widespread among diarrheagenic Escherichia coli in developing countries, where the overuse of antibiotics is common. Information regarding β-lactamases, especially Extended-Spectrum β-Lactamases (ESBLs) in diarrheagenic pathogens should be considered in clinical management when an optimal treatment is needed.. The main objective of this study was to investigate the prevalence of blaCTX-M, blaSHV and blaTEM β-lactamase genes among enteropathogenic E. coli (EPEC) isolates in Tehran, Iran.. Stool specimens were collected from children with diarrhea during a 17-month period from 2011 to 2013. Routine biochemical tests were performed for identification of E. coli isolates. The isolates were further examined by PCR for the presence of eae, stx1, stx2 and bfp genes. EPEC isolates have been screened for different β-lactamase genes. Genotyping EPEC isolates harboring blaCTX-M15 gene was performed through Multi-Locus VNTR Analysis (MLVA).. Of 42 EPEC, eight isolates carried the blaCTX-M1. None of the isolates carried blaCTX-M2 and blaCTX-M9. The blaCTX-M15 variant was identified in all of blaCTX-M1-positive isolates. Furthermore, blaSHV and blaTEM genes were detected in 40.5% (n = 17) and 19% (n = 8) of all EPEC isolates, respectively. No significant association was observed between the existence of bfp gene and presence of those β-lactamase genes (P > 0.05). MLVA analysis revealed high genetic diversity among blaCTX-M15-positive isolates.. Our study emphasized the increasing role of ESBL genes, especially blaCTX-M15 in EPEC isolates..

Lipases (acylglycerol acylhydrolase, E. C. 3. 1. 1. 3) are widely distributed among microorganisms, animals and plants, catalyzing the hydrolysis of triglycerides to free fatty acids and glycerol. Their commercial application includes pharmaceutical, chemical, and paper industries.. This study aimed to isolate and screen lipolytic fungi from coastal waters of the southern Caspian Sea by Internal Transcribed Spacer-Polymerase Chain Reaction (ITS-PCR), and to optimize their lipolytic activity, pH and temperature. The ITS regions possess a high variation among taxonomically distinct fungal species and even within species.. All fungal were tested to determine their lipolytic activity by the Tributyrin agar plate assay. After DNA extraction, lipase-producing fungi were identified via ITS-PCR of rDNA region with ITS1 and ITS4 primers.. Four fungal species were isolated from water samples of the Caspian Sea (north of Iran) between February and June 2011. The nucleotide sequences reported for three of these isolates have been assigned accession numbers from NCBI Gene Bank database. Among these species, Cladosporium langeronii showed maximum lipolytic activity (34 U/mL) and maximum clear zone formation (6 mm) on the Tributyrin agar plates. The optimum pH and temperature for activity were 8.0 and 35°C, respectively.. The findings of this study indicated that these isolates were plant pathogenic fungi, which entered seawater from the environment, and were likely to have a suitable lipase activity on plant oils..

Botrytis cinerea, a haploid Euascomycete fungus that infects numerous crops, has been used as a model system for studying molecular phytopathology. Botrytis cinerea adopts various modes of infection, which are mediated by a number of pathogenicity and virulence-related genes. Many of these genes have not been reported previously.. This study aimed to investigate development and pathogenicity-related genes between a novel nonpathogenic mutant and the Wild Type (WT) in B. cinerea.. Digital Gene Expression (DGE) tag profiling can reveal novel genes that may be involved in development and pathogenicity of plant pathogen. A large volume of B. cinerea tag-seq was generated to identify differential expressed genes by the Illumina DGE tag profiling technology.. A total of 4,182,944 and 4,182,021 clean tags were obtained from the WT and a nonpathogenic mutant stain (BCt89), respectively, and 10,410 differentially expressed genes were identified. In addition, 84 genes were expressed in the WT only while 34 genes were expressed in the mutant only. A total of 664 differentially expressed genes were involved in 91 Kyoto Encyclopedia of Genes and Genome pathways, including signaling and metabolic pathways.. Expression levels of 1,426 genes were significantly up-regulated in the mutant compared to WT. Furthermore, 301 genes were down-regulated with False Discovery Rates (FDR) of < 0.001 and absolute value of log2 Ratio of ≥ 1..

One of the major clinical problems regarding Pseudomonas aeruginosa is attributed to metallo-beta-lactamases (MBL). This group of enzymes is a subset of beta lactamases which belong to group B of Ambler classification and cause hydrolysis of carbapenems. Based on epidemiological studies conducted worldwide, it is proved that prevalence of genes coding MBLs in P. aeruginosa species are different in various geographic zones and even in various hospitals. Therefore, according to the clinical importance of organisms generating MBLs, it is necessary to identify and control these bacteria in hospitals for therapeutic purposes.. The current study aimed to investigate the Metallo-beta-Lactamase VIM-1, SPM-1, and IMP-1 genes among clinical P. aeruginosa species isolated in Zahedan, Iran.. The current study investigated the presence of MBL through phenotypic and genotypic methods and also the pattern of antibiotic resistance in P. aeruginosa species isolated in hospitals. The Minimum Inhibitory Concentration (MIC) against imipeneme was measured for 191 P. aeruginosa species isolated from Zahedan hospitals after identification through biochemical methods and determination of the antibiotic resistance pattern. Strains with MIC > 4 µg/mL were studied by phenotypic and genotypic methods.. The rate of resistance against imipeneme was 5.7% and after carrying out the phenotypic experiments, nine species were identified as of MBL producer. Seven species were confirmed by Polymerase Chain Reaction (PCR) method. Gene VIM-1 was the predominant gene among the positive (antibiotic resistant) species.. The study results showed that MBL genes were present in some of the species isolated from Zahedan hospitals. Regarding the importance of MBL producer bacteria in hospitals, quick identification and evaluation of these clinical species can be considered as an important and basic step for treatment and control of pseudomonad infections..

The periplasmic overexpression of recombinant human interferon beta (rhIFN-β)-1b using a synthetic gene in Escherichia coli BL21 (DE3) was optimized in shake flasks using Response Surface Methodology (RSM) based on the Box-Behnken Design (BBD).. This study aimed to predict and develop the optimal fermentation conditions for periplasmic expression of rhIFN-β-1b in shake flasks whilst keeping the acetate excretion as the lowest amount and exploit the best results condition for rhIFN-β in a bench top bioreactor.. The process variables studied were the concentration of glucose as carbon source, cell density prior the induction (OD 600 nm) and induction temperature. Ultimately, a three-factor three-level BBD was employed during the optimization process. The rhIFN-β production and the acetate excretion served as the evaluated responses.. The proposed optimum fermentation condition consisted of 7.81 g L-1 glucose, OD 600 nm prior induction 1.66 and induction temperature of 30.27°C. The model prediction of 0.267 g L-1 of rhIFN-β and 0.961 g L-1 of acetate at the optimum conditions was verified experimentally as 0.255 g L-1 and 0.981 g L-1 of acetate. This agreement between the predicted and observed values confirmed the precision of the applied method to predict the optimum conditions.. It can be concluded that the RSM is an effective method for the optimization of recombinant protein expression using synthetic genes in E. coli..

Hepatitis C Virus (HCV) infection is diagnosed by antibody and RNA based methods. Patients with anti-HCV sample rate/cutoff rate (S/CO) ratios > 1 are reported as anti-HCV positive. RNA based methods are introduced to confirm positivity in seropositive samples.. The current study aimed to assess relationship between S/CO rates and HCV-RNA levels in the laboratory to identify HCV viremia in patients with a positive anti-HCV..Patients and All serum samples were assayed for anti-HCV by ELISA method. A total of 265 anti-HCV positive patients were tested for HCV-RNA testing by quantitative method using Artus HCV RG Real-time Polymerase Chain Reaction (RT- PCR) kit. Statistical analysis was done by SPSS version 16.. Of the 265 patients with HCV infection, 204 (77%) were male and the mean age was 43.53 ± 13.17 years, ranging 1 - 81 years. No correlation was found between S/CO ratios and HCV-RNA levels. There was significant difference in S/CO ratio between viremic and non-viremic subjects. The sensitivity, specificity, negative predictive value, and positive predictive value were 100%, 81.4%, 100%, and 77.2%, respectively in the S/CO ratio of 2.7.. The present study indicated that anti-HCV S/Co ratio is useful to predict non-viremic patients. A cut-off value of 2.7 can determine the usefulness of HCV-RNA testing. Patients with S/CO < 2.7 are not viremic; therefore, HCV-RNA testing is not recommended. It is suggested that laboratories report S/CO ratio along with anti-HCV results to manage HCV infection better, especially in countries that quantitative HCV testing is expensive or not available..

Tuberculosis (TB) is the world''s second most common infectious disease after Human Immunodeficiency Virus Infection/Acquired Immunodeficiency Syndrome (HIV/AID) and the most frequent cause of mortality especially in developing countries. T regulatory (Treg) cells, which have suppressive activity and express forkhead winged-helix family transcriptional repressor p3 (FoxP3), suppress the immune responses against pathogens such as Mycobacterium tuberculosis. There are controversial results regarding the role of FoxP3 expressing cells in the blood of patients with TB.. The aim of this study was to evaluate the frequency CD4+ CD25+ Treg cells, and FoxP3 and Cytotoxic T Lymphocyte Antigen 4 (CTLA-4) gene expressions in peripheral blood of patients with tuberculosis and patients with positive tuberculin skin test before and after Peripheral Blood Mononuclear Cells (PBMCs) activation with Purified Protein Derivative (PPD)..Patients and In this cross-sectional study, Peripheral Mononuclear Cells (PBMCs) were isolated from peripheral blood of 29 patients with newly diagnosed pulmonary TB and 19 patients with positive tuberculin skin test. The PBMCs were activated with PPD for 72 hours. Activated cells were harvested, RNA was extracted and cDNA was synthesized. A real-time Taqman method was designed and optimized for evaluation of Foxp3 gene expression and SYBR Green method was used and optimized for evaluation of CTLA-4 gene expression. A flow cytometry analysis was used to evaluate the frequency of CD4+ CD25+ Foxp3+ regulatory T cells in both groups.. There was no significant difference in the frequency of CD4+ CD25+ FoxP3+ regulatory T cells between the two groups. Expression of FoxP3 and CTLA-4 in peripheral blood of patients with newly diagnosed TB was significantly lower than the control group after and before activation with PPD.. The expression of FoxP3 and CTLA-4 in PBMCs of patients with newly diagnosed TB was low, which might suggest that Treg cells may be sequestered in the lungs..

Vulvovaginal Candidiasis (VVC) is a frequent, complex and cumbersome condition that can cause physical and psychological distress for the involved individual. Candida albicans was reported as the most common agent of VVC yet it seems that we are recently encountering changes in the pattern of Candida species in VVC.. In this study we assessed different species of Candida isolated from patients with VVC, residing in Sari, Iran..Patients and Two hundred and thirty-four patients with vulvovaginitis were enrolled in this study. Samples were collected by a wet swab. Each vaginal swab was examined microscopically and processed for fungal culture. The identification of Candida species was done by morphological and physiological methods such as culture on CHROMagar Candida media and sugar assimilation test with the HiCandida identification kit (HiMedia, Mumbai, India).. Out of 234 patients with vulvovaginitis, 66 (28.2%) patients showed VVC. Of these patients, 16 (24.2%) had recurrent VVC (RVVC). The age group of 20 - 29 year-olds had the highest frequency of VVC (48.5%). Erythema concomitant with itching (40.9%) was the most prevalent sign in VVC patients. Fifty-seven (86.4%) of the collected samples had positive results from both microscopic examination and culture. In total, 73 colonies of Candida spp. were isolated from 66 patients with VVC. The most common identified species of Candida were C. albicans (42.5%), C. glabrata (21.9%) and C. dubliniensis (16.4%). In patients with RVVC and patients without recurrence, C. albicans and non-albicans species of Candida were frequent species, respectively.. The results of our study showed that non-albicans species of Candida are more frequent than C. albicans in patients with VVC. This result is in line with some recent studies indicating that non-albicans species of Candida must be considered in gynecology clinics due to the reported azole resistance in these species..

The presence of certain oral pathogens at implant sites can hinder the osseointegration process. However, it is unclear how and by what microorganisms it happens.. This study investigated whether the presence of oral pathogens of Porphyromonas gingivalis and Prevotella intermedia individually, play a role in the failure of bone formation by determining the expression profiles of Transforming Growth Factor Beta (TGF-β/Bone Morphogenic Protein (BMP) and Toll-Like Receptor (TLR) pathways in challenged osteoblasts.. Cell viability of P. gingivalis and P. intermedia challenged osteoblasts were determined by WST assay. Changes in osteoblast morphology and inhibition of mineralization were observed by Scanning Electron Microscopy (SEM) and Von Kossa staining, respectively. Expression of TGF-β and TLR pathway genes on challenged cells were identified by RT profiler array. Both P. gingivalis and P. intermedia challenges resulted in reduced viability and mineralization of osteoblasts.. Viability was reduced to 56.8% (P. gingivalis) and 52.75% (P. intermedia) at 1000 multiplicity. Amongst 48 genes examined, expressions of BMPER, SMAD1, IL8 and NFRKB were found to be highly upregulated by both bacterial challenges (Fold Change > 4).. P. gingivalis and P. intermedia could play a role in implant failure by changing the expression profiles of genes related to bone formation and resorption..

High levels of multidrug resistance are usually associated with mobile genetic elements that encode specific resistance genes. Integrons are important genetic elements involved in spreading antibiotic multi-resistance. In special cases, large exogenous segments in bacterial genomes form genomic islands, and one of the functions of these genomic islands is antibiotic resistance. Due to geographical heterogeneity in antibiotic resistance pattern, it is mandatory to determine resistance patterns that are region-specific rather than generalized.. The objective of this study was to detect class 1 and 2 integrons in clinical isolates of Haemophilus influenzae..Patients and Antibiogram tests were carried out for twenty clinical isolates collected from different patients admitted to the Milad hospital. The PCR reactions were performed using universal primers specified for Int1 and Int2 genes attributed to class 1 and 2 integrons. Also amplification of integrase genes related to genomic islands was investigated by designing specific primers.. Of the twenty isolates, all (100%) were resistant to clindamycin, chloramphenicol and tetracycline, 95% to amoxicillin, 50% to ceftriaxone, 45% to ciprofloxacin and 5% to azithromycin. Also, all isolates (100%) were sensitive to trimethoprim/sulfamethoxazole. Class 1 and 2 integrons were not detected in any of the isolates; however the integrase gene attributed to genomic islands was identified in twelve isolates.. Antibiotic resistance gene cassettes may be carried on integron or other genetic elements. The purpose of this study was to detect integron or genomic islands involved in antibiotic resistance profile of the isolates of H. influenzae collected in this study..

Many children become infected with Epstein-Barr virus (EBV) during their childhood. Since the clinical profile of EBV primary infection is challenging, it is important to use the best diagnostic clinical means. Detection of IgM against viral capsid antigen (VCA) by ELISA has been shown to be a reliable method.. This study was conducted to demonstrate the incidence of EBV primary infection, among suspected patients referred to Namazi hospital, Shiraz, Iran..Patients and The sample included 346 patients with an age range of 0 to 20 years (6.31 ± 4.66: 10.97 years). A volume of 5 mL of blood was collected from each case. The patients were divided to four age groups. The sera were tested for the presence of VCA-IgM by commercially available Anti-EBV-VCA ELISA kit.. The results indicated that 104 (30.0%) of the patients were EBV VCA IgM positive, with no significant difference in the incidence of EBV primary infection between males and females. However, the incidence of infection was significantly different between age group I (0 - 5 years) and III (11 - 15 years), and also between age group I (0 - 5 years) and IV (16 - 20 years) (P < 0.05).. Considering the results, accurate and on time diagnosis of EBV primary infection in both children and adolescents will help prevent unnecessary hospitalization, medication and incorrect medical decisions. In addition, this will decrease further treatment costs and related medical procedures..

Gradual development of a useful vaccine can be the main point in the control and eradication of Hepatitis C virus (HCV) infection. Hepatitis C Virus envelope glycoproteins are considered as the main HCV vaccine candidate.. In this study, the Pichia pastoris expression system was used to express a recombinant HCV CoreE1E2 protein, which consists of Core (269 nt-841nt) E1 (842 nt-1417nt) and E2 (1418 nt-2506nt).. By a codon optimization technique based on the P. pastoris expression system, we could increase the rate of recombinant proteins. Moreover, the purified protein can efficiently induce anti-CoreE1E2 antibodies in rabbits, and also by developing a homemade Enzyme-Linked ELISA kit we can detect antibody of HCV Iranian patients with genotype 1a.. In our study, the virus-like particle of rCoreE1E2 with 70 nm size, was shown by Electron microscopy and proved the self-assembly in vitro in a yeast expression system.. These findings of the present study indicate that the recombinant CoreE1E2 glycoprotein is effective in inducing neutralizing antibodies, and is an influential HCV vaccine candidate..

The association between hepatitis C virus (HCV) infection and oral lichen planus (OLP) has been the focus of many studies. Fifteen percent of HCV infections lead to sets of extrahepatic manifestations including lichen planus (LP). The prevalence of HCV is heavily influenced by geographical location.. This study aimed to evaluate the relationship between OLP and HCV infection in Mashhad, northeast of Iran.. Blood samples were taken from 134 OLP patients and 134 healthy controls (without OLP) to screen for anti-HCV by ELISA (third generation) and reverse transcription polymerase chain reaction (RT-PCR) for HCV-RNA.. Of the 134 OLP patients only three (2.23 %) had HCV infection where both anti-HCV and HCV-RNA were positive. All controls were negative for both anti-HCV and HCV-RNA (P = 0.082).. Our investigation illustrated that the prevalence of hepatitis C was higher among OLP patients compared to the control group. These findings are in line with previous results that reported a hepatitis C prevalence of 0.19% among the general population of Mashhad..

The immunomodulative effects of Lactic Acid Bacteria as probiotics have been already demonstrated.. The current study aimed to evaluate the effect of oral administration of Lactobacillus acidophilus on the immune responses and patterns of cytokine production in the BALB/c mice bearing breast cancer.. The current study used thirty inbred BALB/c mice, six- to eight-week-old; they were divided into two groups of 15 each. One group was used as control in each assay. The L. acidophilus (ATCC4356) used in the study was inoculated in MRS broth and cultivated overnight at 37°C under anaerobic conditions, then collected by centrifugation, and re-suspended in Phosphate-buffered Saline (PBS) media. After preparation of the proper amount of the suspension, it was orally administered to the mice via gavage and the control mice received an equal volume of PBS in the same manner.. The results showed that oral administration of L. acidophilus as a potent immunostimulator agent could motivate the proliferation of immune cells. Moreover, it could increase the production of IFN-γ and decrease the production of IL-4, known as Th2 cytokines, in the spleen cell culture. The results showed that the survival time of the L. acidophilus administered mice significantly increased in comparison to that of the control mice.. The current study findings suggested that L. acidophilus can promote immune responses with Th1 bias and may increase the antitumor response. Further, the consumption of this probiotic strain may help to manage the immune response in tumor condition, but more studies are needed to investigate the other mechanisms of this effect..

Adhesion protein E (PE) of Haemophilus influenzae is a 16 - 18 kDa protein with 160 amino acids which causes adhesion to epithelial cells and acts as a major factor in pathogenesis.. In this study, we performed cloning, expression and purification of PE as a candidate antigen for vaccine design upon further study.. At first, the pe gene of NTHi ATCC 49766 strain (483 bp) was amplified by PCR. Then, to sequence the resulted amplicon, it was cloned into TA vector (pTZ57R/T). In the next step, the sequenced gene was sub-cloned in pBAD/gIII A vector and transformed into competent Escherichia coli TOP10. For overexpression, the recombinant bacteria were grown in broth medium containing arabinose and the recombinant protein was purified using metal affinity chromatography (Ni-nitrilotriacetic acid) (Ni-NTA agarose). Finally, the protein was detected using sodium dodecyl sulfate polyacrylamide gel electrophores (SDS-PAG) and confirmed by western blotting.. The cloned gene was confirmed by PCR, restriction digestion and sequencing. The sequenced gene was searched for homology in GenBank and 99% similarity was found to the already deposited genes in GenBank. Then we obtained PE using Ni-NTA agarose with up to 7 mg/mL concentration.. The pe gene was successfully cloned and confirmed by sequencing. Finally, PE was obtained with high concentration. Due to high homology and similarity among the pe gene from NTHi ATCC 49766 and other NTHi strains in GenBank, we believe that the protein is a universal antigen to be used as a vaccine design candidate and further studies to evaluate its immunogenicity is underway..

Vancomycin-resistant enterococci (VRE) are important nosocomial pathogens and food chain has been considered as an assumed source for dissemination of VRE to human.. The presence of VRE isolates from food samples and typing of these isolates with Phene plate, a biochemical fingerprinting method, were investigated.. Thirty samples of meat, chicken and cheese were analyzed for VRE during 2010. Antibiotic susceptibility tests and minimum inhibitory concentration (MIC) were also examined. VRE isolates were typed with the Phene plate system (PhPlate), a biochemical fingerprinting method.. A total of 70 VRE isolates were obtained and identified as Enterococcus faecium by species-specific PCR. All the isolates carried vanA, while none of them harbored vanB. The VRE isolates included 35, 27, and 8 isolates from meat, chicken and cheese, respectively. Typing with the PhPlate revealed a diversity index of 0.78 for E. faecium, containing 10 common and four single types. The results of antibiotic susceptibility and MIC tests showed an increased resistance to ciprofloxacin, erythromycin, ampicillin and gentamicin, to which, 100%, 100%, 100%, and 95% of VRE isolates were resistant, respectively. Only 5% of the isolates were resistant to chloramphenicol and the MIC of the isolates for vancomycin and teicoplanin was ≥ 256 µg/mL and for gentamicin-resistant isolates it was 1024 µg/mL. Conventional and molecular identification tests exhibited that all the isolates were E. faecium carrying vanA. None of the isolates harbored vanB.. The results showed that enterococci are common contaminants in food. Indeed, this study indicates a high prevalence of multidrug-resistant enterococci in food of animal origin in Iran. Isolating some persisting enterococcal isolates revealed that continuous surveillance of antimicrobial resistance in enterococci from food is essential..

The Prevalence of Endocervical Chlamydia trachomatis Infection Among Young Females in Kashan، Iran