International Journal of Molecular and Clinical Microbiology
Volume:9 Issue: 1, Winter and Spring 2019

The incidence of anal squamous cell carcinoma has increased in the last few decades. Anal cancer has been associated with persistent infection with high-risk human papillomavirus (HR-HPV) types. However, only a few data reflect the status of anal HPV infection in women with cervical neoplastic and paraneoplastic lesions. The objective of the present study was to investigate the distribution of HPV genotypes and abnormal anal cytology in sexually active women with cervical disease from Bogota, Colombia. We therefore performed anal cytology for type-specific HPV identification in 134 anal samples of sexually active women from Bogota, Colombia, presenting cervical disease; using a commercial molecular technique (Linear Array®; Roche, Molecular Systems, USA). Results of anal cytology were normal in 93.3% of samples, while the remaining 5.2% was classified as atypical squamous cells of undetermined significance (ASC-US), 0.75% high-grade anal intraepithelial neoplasia (LGAIN-II/III,) and 0.75% low-grade AIN ( (HGAIN-I). The analysis for viral infection in the 134 anal samples showed HPV in 61.5% of the studied population. In general, higher HPV infection values were seen in more compromised anal lesions. The most prevalent viral genotypes were HPV-16 (27.7%), -6 (25.30%), -58 (15.70%), -18 (9.60%) and -53 (9.60%). Multiple infections were more common than single ones, and HR-HPV genotypes were present in 69.80% of positive samples. Updating data concerning the distribution of genotypes that infect the anal mucosa may contribute to the implementation of strategies focused on reducing the rate of anal cancer morbidity and mortality.

This study was undertaken to select and analyze epitopes of Corynebacterium diphtheriae toxin using online and offline computerized analysis methods, after that affinity of MHC epitopes to HLA-DRB1*0101 was analyzed by Hex-protein protein software. The B cell epitopes were predicted via Immune Epitope Database (IEDB) and MHC class II epitopes were predicted by Vaxign softwarer. A physicochemical analysis of candidate vaccine revealed that the designed vaccine has a molecular weight of 59.062 kD. The estimated half-life of candidate vaccine was found to be greater than 30 hours (mammalian reticulocytes, in vitro), greater than 20 hours (yeast, in vivo), and greater than 10 hours (Escherichia coli, in vivo). The instability index (II) is computed to be 31.31 (<40), the aliphatic index was found to be 65.80, and the vaccine was considered stable. The grand average of hydropathicity was -0.580, therefore, the vaccine is a hydrophilic protein and probably it interacts with molecules of water. The result of allertop, toxinpred showed that the vaccine is non-allergenic and non-tixic. According to the obtained data from protparam and pepcalc, our designed vaccine is soluble and no transmembrane helix was projected, hence no expression difficulties are anticipated in the development of the vaccine. The membrane helices value of vaccine was 33.64%. The result of protein protein docking analysis showed that maximum affinity of candidate vaccine to HLA-DRB1*0101 with the score of -636.85. The result of this study showed that the candidate vaccine can be stimulate HLA-DRB1*0101.

A large amount of feather is produced each year in the poultry industries which is buried in landfill or burned in the power plant generator. Feathers are purified keratinous proteins that can be degraded by bacteria and fungi. The aim of this study was to isolate and identify keratinase producing bacteria from the feather waste dumping sites around Gorgan district, Golestan province, Iran. Soil samples were collected from six poultry farms and slaughterhouses. The keratinolytic bacteria were isolated and identified based on feather and keratin degradation in feather meal broth and keratin agar. Isolates that showed more keratinolytic activity were identified using morphological, biochemical, and molecular tests. Then, the effect of pH and temperature was examined on the growth of the isolates. Based on the results, keratinase-producing isolates belonged to the Bacillus genus, among the isolates, the most keratinolytic activity was related to Bacillus megaterium strain SR7 and it showed the best growth at pH 6.8 and 37°C in feather meal broth and keratin agar. The results of the present study were consistent with the results of many similar studies and found that Bacillus strains are important producers of keratinase enzyme and therefore promising organisms for the management of chicken feather wastes through efficient biodegradation.

Antibacterial activity against gastrointestinal bacteria, tolerance to acidic conditions and bile salts are the main indicators of probiotics. The aim of this study was isolation of lactic acid bacteria with probiotic potential of bovine colostrum. Colostrum samples were cultured on selective culture. Identification of lactic acid bacteria carried out by biochemical tests of fermentation of sugars, growth at 10 and 45°C and 6.5% salt. Antagonistic effect of culture supernatant of isolates against native isolates S. aureus, E. coli, P. aeruginosa, B. cereus and standard strains E. coli, P. aeruginosa, S. dysentriae, S. aureus, B. cereus and E. faecalis carried out by well method and tolerance of isolates to acidic conditions and bile salts were evaluated. L. plantarum was the most species of isolated lactic acid bacteria (36.32%). Culture supernatant of L. plantarum more than other isolates was showed antibacterial activity. None of the isolates were able to grow at pH 2, but L. plantarum, L. casei, L. acidophilus, S. thermophilus and E. faecalis were able to tolerate pH of 3. Also, L. plantarum, L. casei, E. faecalis and E. faecium were able to grow in presence of 0.3 and 1% bile salt. Colostrum as a traditional dairy food can be considered as a very suitable reservoir for isolation of lactic acid bacteria with probiotic potential. Due to significant antibacterial activity of L. plantarum and the ability to grow in acidic condition and various concentrations of bile salts, probiotic potential of this isolate was evaluated as desirable.

Stored nuts in the markets are naturally infected by different fungal species such as Aspergillus. The present study was carried out to evaluate the occurrence of toxicogenic A. flavus strains on nuts in Iraq. A total of 112 nuts samples including hazelnuts, pistachio, peanut, and walnut with typical symptoms of dark and green discolored lesions on kernels were collected from various markets in Baghdad. Strains of Aspergillus spp. isolated from nuts seeds and their morphological characterization was based on MEA, PDA, and CYA. The identification of A. flavus isolates were confirmed molecularly using primers T1/T2. A total of 25 fungal isolates belonged to Aspergillus species that were identified as A. niger (10), A. flavus (10), and A. japonicus (5). In molecular analysis, sequences of partial β-tubulin gene were blasted in GenBank to confirm morphological identification of A. flavus isolates. Aflatoxins (AFs) contamination of ten infected samples with A. flavus was evaluated using ELISA method. Natural occurrence of AFs could be detected in all tested samples, ranging from 6.50–74.48 μg/kg. Our results completed the previously data about genetic potential of AFs production of A. flavus strains in Iraq and revealed in infected nuts also can be one of the great concern in Iraq.

This clinical trial study was performed on 144 patients suspect to have otitis externa between 2017 and 2018 in the ENT clinic of Ayatollah Rouhani hospital in Babol. For the treatment on the same day, the patients were categorized into two groups randomly. In the first group, after suction clearance, ciprofloxacin powder and betamethasone ointment were used, while in the second group after suction clearance, ciprofloxacin drop and betamethasone drop were used in the ear canal. All patients reevaluated after one week. After preparation of the results of culture of the samples, 83 individuals including 44 in the first group (ciprofloxacin powder and betamethasone ointment) and 39 subjects in the second group (ciprofloxacin drop and betamethasone drop) had bacterial otitis externa. The mean intensity of pain in the first and second groups was 2.61±2.13 and 2.8.00, respectively which showed a significant difference (Pvalue<0.001). The itching was 38 (86.4%) and 6 (15.4%) cases in the first and second groups, respectively, which showed a significant difference (Pvalue<0.001). The discharge in the first and second groups was 10 (22.7%) and 1 (2.6%) respectively which again had a significant difference (Pvalue=0.007). The extent of diminished hearing in the first and second groups was 17 (38.6%) and 0 (0%), respectively, indicating a significant difference (Pvalue<0.001). Finally, inflammation in the first and second groups was 20 (45.5%) and 0 (0%), showing a significant difference (Pvalue<0.001). This study indicated use of ciprofloxacin powder and betamethasone ointment is less effective than applying ciprofloxacin drop and betamethasone drop.

Bronchopneumonia caused by Mannheimia haemolytica and Pasteurella multocida, in sheep and goats often occurs in livestock that are under stresses such as transportation, milk catching, changing diets, or being with other animals from other farms. In this study, from June to November 2018, 230 goats at a slaughterhouse in Rasht were examined for pulmonary lesions. Of the total 230 samples, 82 cases (35.6%) had one or more macroscopic lesions. Mannheimia haemolytica, Pasteurella multocida and Bibersteinia trehalosi were detected in 0.9%, 0.45% and 0.45% of positive samples, respectively. In two cases (0.9%), bronchiectasis was observed in the lower anterior region. In the microscopic view of the tissue sections, with the exception of one case that had typical symptoms of acute interstitial pneumonia and oxidative phase observed, 81 other cases (35.2%) showed typical symptoms of enzootic pneumonia and, in one case, focal necroses were observed. The inflammation of the pleura was not seen in goats less 6 months of age, but was observed in 44% of samples 6-12 months and 95% of samples more than 12 months. Based on this study that was conducted for the first time in the north of Iran, enzootic pneumonia and inflammation of the pleura are the most important problems of goats over one year and Mannheimia haemolytica can be one of the potential causes of pneumonia.

Streptomyces are capable of producing secondary metabolites including antibiotics and the main environment of these microorganisms is soil. The purpose of this study was to isolate and evaluate Streptomyces producing antibacterial compounds from different parts of east of Gilan province and optimizing antibacterial compounds produced by them. In this study, after isolation and purification of Streptomyces, to investigate its antibacterial activity against tested pathogenic microorganisms including Micrococcus luteus PTCC 1408, Bacillus cereus PTCC 1154, Staphylococcus aureus PTCC 1189, Pseudomonas aeruginosa PTCC 1310, Salmonella typhi PTCC 1609 and Proteus mirabilis PTCC 1776 were investigated. Streptomyces was identified based on morpholigaical, biochemical, physiological, 16SrRNA gene sequencing and phylogenetic analysis. Optimization of various factors in the content of production of antibacterial compounds were investigated. In this study, 16SrRNA gene sequencing revealed that the isolate belonged to Streptomyces genus and with the highest similarity (95.70%) to Streptomyces malachitospinus indicating significant differences at species level and it can be introduced as a new species. This isolate showed significant activity against pathogenic microorganisms and in relation to optimizing the maximum production of optimal antibacterial compounds in culture medium, pH, temperature, carbon and nitrogen sources and incubation time were ISP2, 7, 28 0C, glycerol, yeast extract and 7 days, respectively.The results show that the soils of the eastern regions of Gilan province are a rich source of antimicrobial compounds, which due to the resistance of the pathogenic microorganisms to antimicrobial drugs, is essential for the production of antimicrobial compounds of natural origin.

Azole compounds have been a treatment option for Candida glabrata infections. However, azole resistance can occur through different mechanisms such as alterations in ERG11 (lanosterol 14α-demethylase). Aspirin (ASA), a non-steroidal anti-inflammatory drug, showed antimicrobial activity against Candida. The purpose of this study was to evaluate the synergistic effect of ASA as an anti-inflammatory drug with the fluconazole on the azole-resistant C. glabrata isolates. In the cross-sectional study, a total of 60 oral samples were collected from the school of dentistry, Tehran University of medical sciences. After confirmation of fungal isolates, template DNA was extracted and PCR for detection of Erg11 gene by specific primers was performed in all Fluconazole-resistant isolates. MIC for ASA was determined using broth dilution method in 96‐well plates. RNA was extracted and cDNA synthesized according to the Omniscript RT kit instructions. The effect of ASA in the treated and non-treated groups on ERG11 gene expression was determined by Real-Time PCR technique. Out of 60 collected samples, 12 (20%) C. glabrata were isolated. All of these isolates were resistant to fluconazole and carried ERG11 genes. Real-time PCR results showed that the combination of ASA with fluconazole reduced ERG11 gene expression. It is concluded that, treatment of candidal infections with ASA significantly reduced resistance in to azole compounds by down expression of ERG11 gene, suggesting that an NSAID might be useful for azole-resistance candidal infections.

The genus Malassezia consist of lipophilic yeasts are known to be as component of the normal microflora of human skin and many mammals and birds. The purpose of this study was determinate the distribution of Malassezia sp. on the horses skin in the in the north of Iran and identification of them according to horses’ sex ,age,breed and living geographically area. During the 15 month period, sampling was carried out using the scraping methods from different areas of skin; 256horse. A part of samples surveyed microscopically with methylene blue stain methods. All samples were inculcated onto sabouraud glucose agar media supplement with olive oil. The identification to lipid – dependence yeasts was based on the ability to use certain Tweens.The cremophore El assimilation test and splitting of Esculin were used as additional key characters. Other tests such as the catalase reaction and the morphological characteristics on SGA was administered. The prevalence of Malassezia spp. in the studied horses was 33.98%. M.pachydermatis was only isolated in 12 horse whereas lipid-dependent species isolated in 75 horse. The most frequently isolated Malassezia spp. was in groin with24.27%,and the least of it, dorsrum 15.54%.The most frequently isolated species were M. globosa (22.33%) followed by M. fur fur (18.45%) ,M. restricta (11.59%), M.pachydermatis (11.65%) ,M.obtusa (11.65%) , M. sympodialis (11.65%) and M.slooffiae (10.69%). Distribution of Malassezia species in the healthy body have been investigated using culture-based and biochemistry techniques, and variable results have been reported from different geographical regions