دو ماهنامه میکروب شناسی پزشکی ایران
سال هفتم شماره 4 (زمستان 1392)

The uropathogenic Escherichia coli as a most common cause of urinary tract infections can transmit to the reproductive system. Their various virulence factors cause adhesion, invasion and damage to reproductive epithelium. This study was carried out in order to detection of virulence factors of uropathogenic Escherichia coli isolates from infertile women high vaginal swabs. Totally, 70 and 30 high vaginal swabs were taken from infertile and fertile women, respectively. All swabs were cultured and the positive samples were studied for the presence of uropathogenic virulence genes using PCR method. Thirteen out of seventy swabs samples were positive for E. coli (18.57%). There were no E. coli bacteria in fertile women. Eight of the positive samples were from infertile women with the history of urinary tract infections (61.53%). The most commonly detected virulence genes were papGI (84.61%), set-1 (76.92%), papGII (61.53%), papGIII (53.84%) and sen (53.84%). The high vaginal Escherichia coli harbored certain virulence genes of uropathogenic Escherichia coli strains. The urinary tract infections should be treated well to diminish its upstream transfer into vagina. Some more investigation should be perform for identifying the epidemiological aspects of uropathogenic Escherichia coli in high vaginal part of infertile women.

The ability of adherence to the host cell surface, present of sidrophore receptors and α-hemolysin (hlyA) are common virulence attributes produced by Escherichia coli that enhances virulence in a number of clinical infections. It has been revealed that E. coli strains that infect uro-genital tracts have different virulence factors. This study was aimed to determine the prevalence of genes encoding virulence factors in cervico-vaginal pathogenic E. coli isolated from women attending Gynecology clinics in Zabol-Iran. In this study, 380 cervico-vaginal swabs were obtained from patients with genital tract infections referred to Gynecology clinics in Zabol-Iran during the period (from January to July 2013). A total 132 E. coli isolates were confirmed by conventional biochemical tests. DNA was extracted from all isolates by the boiling method and then DNA was used to determine the presence and prevalence of selected virulence genes including hlyA, iroN, iucD and fimH. The phylo-groups of isolates were determined by detection of yjaA and chuA genes and fragment TspE4.C2. In 132 samples, the frequency of hlyA, iroN, iucD and fimH genes were 71, 37, 31 and 17% respectively. Four major phylogenetic lineages (A, B1, B2, and D) were found in 7%, 19%, 62% and 14% of isolates, respectively. Isolates present in group B2 showed highest presence of virulence genes. This results show that the iroN, iucD, and fimH genes were the most virulent genes of E. coli isolates obtained from patients with uro-genital tract infection. These findings can be valuable in etiology of cervico-vaginal infections (CVIs), CVI administration and management and success of treatment strategies.

Salmonellae infection is one of the most important foodborne diseases. Antimicrobial drug resistance is increasing among Salmonella spp. and causes significant therapeutic problems in the treatment of diseases caused by this organisms. The aim of this study was to investigate the prevalence of class 1 integrons in Salmonella enterica and their relationship with antimicrobial resistance in clinical isolates from Iran during 2007-2009. Salmonella spp. strains were isolated from several hospitals in Tehran, Iran. The isolates were identified by standard biochemical tests and agglutination using specific antisera. The susceptibility of the isolates was determined according to the CLSI guidelines. Class 1 integrons were detected by PCR. Statistical comparisons of the frequency of resistance between integron-positive and integron-negative were made using Pearson χ2 test or the Fisher’s exact test. A P≤0.05 was intended as the level of statistical significance. Class 1 integrons were detected in 39.1% of the strains. Integron-positive isolates represented seven different Salmonella enterica serotypes. All Salmonella isolates carrying class 1 integrons showed multiple drug resistance. Our findings showed that integrons class 1 were widely distributed among Salmonella enterica isolates. Surveillance and monitoring of antimicrobial drug resistance, including screening for integrons as likely indicators of drug resistance and acquisition of new resistance traits, are necessary steps in planning effective strategies for containing this phenomenon within foodborne infection organisms.

PprI is one of newly gene identified in Deinococcus radiodurans and plays a critical role in DNA repair and protection against ultraviolet radiation stress. The aim of this study was to clone, express and characterize PprI gene in Escherichia coli. The PprI gene of D. radiodurans was constructed in pGEM-B1 vector and the cloned gene was subcloned into pET21a expression vector. The pET21a-PprI was expressed in E. coli (Origami strain). Expression of recombinant PprI was confirmed by SDS-PAGE gel electrophoresis and western blotting. In addition, the UV-C radiation resistance of recombinant E. coli was determined. The chimeric pET21a plasmid containing the C -terminal fusion of His-tag with PprI gene was successfully constructed and the medium condition for the expression was optimized. In addition, transformed E. coli had higher resistance to radiation than the original strain. The results of this study indicated that PprI protein could be a potential candidate to be considered as a radioresistant protein. However, the UV-C radioresistance potency of recombinant PprI should be analyzed in prokaryotic and eukaryotic systems

Streptomyces clavuligerus is one of the most important strain that produce clavulanic acid that wildly used in combination of strong but sensitive to β-lactamase antibiotics in clinics. The cas2 is one of the important genes in the biosynthesis pathway of clavulanic acid. The recombinant construct pMTcas2 which contain cas2 gene is obtained from Isfahan University. Recombinant plasmid extracts from streptomyces lividans and confirm by enzyme digestion. The streptomyces clavuligerus protoplast was prepared and transformation was done by using polyethylene glycol. Transformation was confirmed by plasmid extraction and PCR using cas2 specific primers. Finally, bioassay method was used to survey the effect of extra copy of cas2 on clavulanic acid production. Plasmid extraction was initially carried out and the structure of plasmid was confirmed by digestion. The typical white colony was seen on protoplast recovery culture containing thiostrepton antibiotic and gray spores were detected after one week. Plasmid extraction was done from transformed strain and transformation was confirmed by PCR. The results of the bioassay show that amplification of the cas2 gene in multicopy plasmids resulted in a 4.1 fold increase in clavulanic acid production. The bioassay was done and the diameters of zone of inhibition in control and sample were compared. The results of the bioassay show that amplification of the cas2 gene in multicopy plasmids resulted in a 4.1 fold increase in clavulanic acid production. Overproduction of clavulanic acid decreases the cost of its dependent drug production.

Protease are the most important industrial enzymes. The %60 of industrial enzymes sale belongs to proteases. Alkaline proteases have many applications in industry. The main purposes of this study was isolation and identification of alkaline protease producer yeasts and investigation of different conditions effects on producers enzyme production isolation. Method and material: The wastewater samples of several food manufacturies in Isfahan were collected. For screening of the best alkaline protease the enzyme assay, Lowry method was performed, and the effects of different culture conditions on the production of alkaline proteases were determined. The DNA extraction and PCR were performed. Among the isolates strains, Candida viswanathii was selected for further processing. The maximum production of enzyme was obtained in a culture medium containing %0.5 NaNO3, initial pH 8.0 and aeration speed of 200 rpm after 72 hours incubation at 30 °C. Due to the high demand for industrial enzymes in the Country and the high activity of alkaline proteases produced by strain. It seems that the native strain can achieve high production of alkaline proteases.These native strains could be resulted in the independence of our country in industrial enzymes production.

Dermatophytosis is a common contagious disease caused by fungi known as dermatophytes. Dermatophytes belong to a group of organisms that are able to break down the keratin in tissues such as the epidermis, hair and nails. In this survey we examined fungal contaminations of Iranian coins and paper bills obtained from tinea patients. A total of 600 papers and 100 coins were examined. Of the 600 paper bills tested, 400 were obtained from tinea patients,100 were from general population, and 100 were new un-touched bills. The paper bills were cultured on Scc medium (1 gr/lit)containing cycloheximide according to aseptic condition. One Scc plate was used for each bill. The coins were also cultured on the same Scc media by both direct contact as well as scotch tape method. 6 dermatophytes (1.6%) were obtained from the 400 paper bills from tinea patients). Of these six, 2 were E.floccosum and 4 were T.mentagrophytis. The money of tinea patients, are able to transfer the disease to general population but there is not enough sensitivity, to take money from these patients. So we must take appropriate strategies to prevent these contaminations.

Urinary tract infection (UTI) is a common infection. Having enough knowledge about the etiology of UTIs and also antibiotic resistant pattern of E.coli as the commonest uropahtogen helps the physicians to deal with these kinds of infections. The aim of this study is to determine the prevalence of uropathogens originated by bacteria and antibiotic resistance pat-tern of E.coli as the most common cause of urinary tract infections. From 2010/09/23 to 2011/09/23 all medical case records of all patients diagnosed with UTI at Shahid Faghihi hospital (Shiraz, Iran), Shahid Motahari hospital (Marvdasht, Iran) and a private pathology laboratory (Saadat-Shahr, Iran) were analyzed in order to assess the prevalence of all bacterial uropathogens and the direct effect of seasonal changes on it. To determine the antibiotic resistance pattern of E.coli, from 2011/07/19 to 2011/08/30, in a cross-sectional study, 100 E.coli out of 146 gram-negative bacteria were detected by biochemical and gram stain tests. Antibiotic resistance tests were done using CLSI criteria. Results and The prevalence of staphylococcus and Acinetobacter in both sexes and the prevalence of Streptococcus, Klebsiella and Diphtheroids among females were altered by seasonal changes. The resistance rates detected were 1% to Amikacin, 98% to Tetracycline, 15% to Gentamicin, 72% to co-trimoxazole, 56% to Ciprofloxacin, 66% to Nalidixic acid, 17% to Nitrofurantoin. In addition to the patients’ gender and the region of study, seasonal changes fol-lowed by thermal and humidity changes, is another significant factor which influences the etiolo-gy of UTIs. Also antibiotic resistance pattern would be different even in neighbor cities.