Jundishapur Journal of Microbiology
Volume:8 Issue: 6, Jun 2015

Food-borne pathogens are among the most significant problems in maintaining the health of people. Many probiotics have been widely reported to alleviate and protect against gastrointestinal infections through antibacterial secretion. However, the majority of them cannot always play antagonistic roles under gut conditions. Probiotic bacteria of human origin must possess other protective mechanisms to survive, out-compete intestinal flora and to successfully establish in their new host at a significant level. Probiotic characteristics of Lactic Acid Bacteria (LAB) and bifidobacteria isolated from the feces of Thai infants were primarily investigated in terms of gastric acid and bile resistances, antibacterial activity and mucin adhesion ability. Antagonistic interaction through secretion of antibacterial compounds and competitive exclusion against food-borne pathogens were also evaluated. Culturable LAB and bifidobacteria were isolated from feces of Thai infants. Their ability to withstand gastric acid and bile were then evaluated. Acid and bile salt tolerant LAB and bifidobacteria were identified. They were then further assessed according to their antagonistic interactions through antibacterial secretion, mucin adhesion and competitive mucin adhesion against various food-borne pathogenic bacteria. Gastric acid and bile tolerant LAB and bifidobacteria isolated from healthy infant feces were identified and selected according to their antagonistic interaction against various food-borne pathogenic bacteria. These antagonistic probiotics included four strains of Lactobacillus rhamnosus, two strains of L. casei, five strains of L. plantarum, two strains of Bifidobacterium longum subsp. longum and three strains of B. bifidum. All strains of the selected LAB inhibited all pathogenic bacteria tested through antibacterial secretion, while bifidobacteria showed high level of competitive exclusion against the pathogenic bacteria. These human-derived LAB and bifidobacteria exhibited different mechanisms involved in pathogenic inhibition. Therefore a combination of these probiotic strains could be a great promise and possibility for the development of probiotic products to effectively prevent and control food-borne infection in humans.

Dermatophytes are a group of keratinophilic fungi worldwide, which can infect the skin, hair and nails of humans and animals. This genus includes several species that present different features of dermatophytosis. Although, laboratory diagnosis of dermatophytes is based on direct microscopy, biochemical tests and culture, these manners are expensive, time consuming and need skilled staff. Therefore, molecular methods like PCR-RFLP are the beneficial tools for identification, which are rapid and sensitive. Thus, dermatophyte species are able to generate characteristic band patterns on agarose gel electrophoresis using PCR-RFLP technique, which leads to successful identification at the species level within a 5-hour period. The purpose of this study was to study inter- and intraspecific genomic variations for identification of clinically important dermatophyte species obtained from clinical specimens in Isfahan, Iran using PCR-RFLP. From March 2011 to August 2012, 135 clinical isolates were collected from infected patients at Isfahan, Iran. ITS1-5.8S-ITS2 region of rDNA was amplified using universal fungal primers. Subsequently, amplified products were digested by the MvaI restriction enzyme. Using discriminating band profiles on agarose gel, dermatophyte species were identified. However, DNA sequencing was used for unidentifiable strains. The specimens were obtained from skin scrapings (70.3%), nail (24.4%) and hair (5.1%) clippings. Most patients were between 21 - 30 years and the ratio of male to female was 93/42. Trichophyton interdigitale was the commonest isolate (52.5%) in our findings, followed by Epidermophyton floccosum (24.4%), T. rubrum (16.2%), Microsporum canis (2.2%), T. erinacei (1.4%), T. violaceum (1.4%), T. tonsurans (0.7%) and M. gypseum (0.7%) based on PCR-RFLP. Combination of traditional methods and molecular techniques considerably improves identification of dermatophytes in the species level in clinical laboratories, which can lead to properly antifungal therapy and successful management of infections. However, restriction and specificity and sensitivity should be lowered and increased, respectively, to be useful for a wide variety of clinical applications..

Although studies focused mainly on the identification of periopathogenic bacteria, recent reports have suggested that various herpes viruses may also be involved in the occurrence and progression of different forms of periodontal diseases. This study aimed to compare the prevalence and load of Epstein-Barr Virus (EBV) and Human Cytomegalovirus (HCMV) in subgingival tissue specimens between chronic periodontitis and healthy sites.Patients and A total of 60 samples from the systematically healthy patients with chronic periodontitis participated in this study (mean age, 35 ± 7). Clinical periodontal evaluation included the plaque index (PI) (Loe and Silness), bleeding on probing (BOP) (O’Leary), bleeding index, periodontal pocket depth (PPD) and clinical attachment level measurement. Tissue specimens harvested from > 6 mm periodontal pockets and from ≤ 3 mm sulcus depth in a quadrant of the same patient using periodontal curettes. Moreover, the unstimulated whole saliva was gathered as a shedding medium. A Taq-man Real-Time Polymerase Chain Reaction assay was used to identify genomic copies of periodontal HCMV and EBV. Data were analyzed by the Wilcoxon-signed ranks and Friedman tests using the SPSS 16 software. Out of 60 samples of subgingival tissues taken from the patients with chronic periodontitis, EBV count was the highest in saliva and the least in the tissue sample with PD < 3 mm (P < 0.05). The highest HCMV count was in saliva and tissue samples with PD > 6 mm (P < 0.05). According to the results of this study, quantification of HCMV and EBV observed in this study is high in periodontal tissue samples of severe chronic periodontitis.

Acinetobacter calcoaceticus baumannii (ACB) complex are Gram-negative opportunistic bacteria with low virulence properties. Their resistance to antibiotics has become a matter of concern in hospital infections. The present study aimed to determine the prevalence and antimicrobial susceptibility of ACB isolates collected from the Nemazee hospital of Shiraz. In addition, Pulsed Field Gel Electrophoresis (PFGE) was used to determine the genetic patterns of these strains.Patients and In this cross-sectional study, 93 strains of ACB complex were isolated from patients of Nemazee hospital, Shiraz, Iran. The antibiotic susceptibility patterns of the isolates to the following 15 antibiotics were determined: gentamicin, ticarcillin, ceftazidime, co-trimoxazole, imipenem, piperacillin tazobactam, amikacin, aztreonam, sulbactam, meropenem, tobramycin, cefotaxime, ceftriaxone, colistin, polymyxin B. Pulsed Field Gel Electrophoresis was used to determine the clonal relationship of these strains. Most of the isolates were found to be resistant to cefotaxime, co-trimoxazole, ceftriaxone, aztreonam, ceftazidime and ticarcillin (90%), and the least resistance was observed to colistin and polymyxin B. Among the 93 tested samples, 35 antimicrobial susceptibility patterns and 47 PFGE patterns were obtained. High resistance to antibiotics was observed among the strains of ACB complex and the least resistance was towards colistin and polymyxin B, indicating that these antibiotics could be effective for treatment, in case there is no other choice. Using PFGE, the similarity between some strains of Acinetobacter was determined, which indicated epidemics in different parts of the hospital; such epidemics can in turn lead to increased incidence of Acinetobacter infections.

Asthma is a chronic inflammatory air-way disease with increasing prevalence rate during the recent years. There are studies about the relationship between asthma and infectious diseases, including the association between asthma and Helicobacter pylori. According to the latest studies, there is an epidemiological correlation between asthma prevalence and prevalence of H. pylori. The aim of this research was to study the correlation between H. pylori and asthma by biopsy in five to eighteen year-old children who had undergone endoscopy at Shahid Beheshti Hospital.Patients and Three hundred children (5 to 18 years old) undergoing endoscopy owing to gastro-intestinal problems at Shahid Beheshti Hospital were observed for childhood asthma using the Gina 2010 questionnaire which included 24 questions with “yes” and “no” answers to identify asthmatic patients with five positive answers. Next, the patients were referred to an allergy and asthma specialist for clinical examinations, spirometry and post bronchodilator test (Post BD). Among 138 H. pylori positive patients, eight cases (5.8 %) were asthmatic while of the 162 H. pylori negative patients 28 (17.3%) were asthmatic. This difference was statistically significant (P Value = 0.002). The correlation between H. pylori and asthma was studied after controlling the confounding variables including, gender, age and family history. The results obtained for the above-mentioned variables were significant (P Values of 0.004, 0.005 and 0.002, and Odd-Ratio Mantel Haenszel (ORMH) of 3.38, 3.24 and 4.06, respectively). Our findings showed that there is an inverse correlation between H. pylori and asthma. Performing more studies with larger sample sizes is necessary to confirm these results.

Treatment of Pseudomonas aeruginosa PAO-1 infections through immunological means has been proved to be efficient and protective. The purpose of this study was to produce a conjugate vaccine composed of detoxified lipopolysaccharide (D-LPS) P. aeruginosa and diphtheria toxoid (DT). Firstly, LPS was purified and characterized from P. aeruginosa PAO1 and then detoxified. D-LPS was covalently coupled to DT as a carrier protein via amidation method with adipic acid dihydrazide (ADH) as a spacer molecule and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDAC) as a linker. The molar ratio of LPS to DT in the prepared conjugate was 3:1. The immunogenicity of D-LPS-DT conjugate vaccine in mice model was evaluated as well. The conjugate was devoid of endotoxin activity and 0.125 U/mL of D-LPS was acceptable for immunization. D-LPS-DT conjugate was nonpyrogenic for rabbits and nontoxic for mice. Mice immunization with D-LPS-DT conjugate vaccine elicited the fourfold higher IgG antibody compared to D-LPS. Anti-LPS IgG antibody was predominantly IgG1 subclass and then IgG3, IgG2a and IgG2b, respectively. Vaccine based on the conjugation of P. aeruginosa PAO-1 LPS with DT increased anti-LPS antibodies and had a significant potential to protect against Pseudomonas infections.

Human T-cell lymphotropic virus types Ι and ΙΙ (HTLV-Ι and HTLV-II) are deltaretroviruses which may cause leukemia, lymphoma and HTLV-associated myelopathy/tropical spastic paraparesis (HAM/TSP). In addition, HTLV-1 may be related to thalassemia and hemophilia cases after blood transfusion. The aim of this study was evaluation of the prevalence of HTLVs in patients with hematological disorders (leukemia, thalassemia, lymphoma and hemophilia).Patients and This cross-sectional study was conducted during April to October 2012. A total of 101 serum samples were collected from patients and were stored at -20ºC. DNA was extracted from serum by an extraction kit. The extracted DNA was tested by polymerase chain reaction (PCR) for detection of HTLV-Ι and HTLV-II pol and tax gene sequences, respectively. Samples were collected from 67 (66.33%), 20 (19.80%), 4 (3.96%), and 10 (9.90%) patients with thalassemia, leukemia, lymphoma and hemophilia, respectively. One thalassemia sample was HTLV-Ι positive, but none of the samples contained the genome of HTLV-II. The prevalence of HTLV-Ι in this study in patients with hematological disorders was 0.99%. The prevalence of HTLV-Ι in hematological disorders was similar to that of other parts of Iran. The present study revealed that HTLV-Ι screening should be performed before blood transfusion to reduce the risk of virus transmission in patients with hematological disorders. More study should be performed to detect these viruses in blood donors.

Glucose is the preferred carbon and energy source in most organisms and plays an active role in the regulation of many biological processes. However, an excess of glucose leads to such undesirable conditions as diabetes and age-related diseases. Since Schizosaccharomyces pombe homologous of many human genes, it offers several advantages for the investigation of the molecular mechanisms underlying human disease and aging studies. We have identified two glucose-repression-resistant mutants (ird5 and ird11) of S. pombe. We aimed to investigate the possible relationship between lifespan extension and oxidative stress response induced by exposure to hydrogen peroxide alongside the trehalose accumulation level by using the two S. pombe mutants (i.e. ird5 and ird11), which are repressed by glucose and are resistant to oxidative stress. We employed trehalose accumulation measurement and colony-forming unit (CFU) counting using the ird mutants in exponential and stationary phases and compared them to the wild type grown in repressed, de-repressed, and stressed conditions to clarify the possible relationship between glucose signaling, oxidative stress response, and lifespan in S. pombe. The lifespan of the ird5 mutant was significantly longer that of either the ird11 mutant or the wild type cells. Under repressed condition, the trehalose content was increased remarkably on the 3rd day of the study in the ird11 mutant and the wild type. Under de-repressed condition, the level of intracellular trehalose was notably increased on the 3rd day in ird11. Under stressed condition, the trehalose level in ird11 was increased on the 3rd day as a pattern similar to that observed in the wild type. Our results demonstrated no significant correlation between the ird5 lifespan and the trehalose concentration. Likewise, the correlation between lifespan extension, trehalose accumulation, and cellular resistance to hydrogen peroxide was not significant.

Proteus mirabilis is an important uropathogen that causes complicated Urinary Tract Infection (UTI) and induces diarrhea in infants. This study aimed to investigate P. mirabilis infection in newly weaned infant rhesus monkeys (Macaca mulatta) and ferrets (Mustela putorius furo) with diarrhea. Stool samples were collected from 74 rhesus monkeys and 12 ferrets with diarrhea. Proteus mirabilis was isolated from the samples through Polymerase Chain Reaction. The isolated P. mirabilis was subjected to antimicrobial susceptibility tests. Seven (7/74, 9.5%) and four (4/12, 30%) P. mirabilis strains were detected in the stool samples collected from the monkeys and ferrets, respectively. Sequence analyses showed that the isolated P. mirabilis was closely related to P. mirabilis strain HI4320, which was isolated from the urine of a patient with a long-term indwelling urinary catheter. In addition, the isolates demonstrated multidrug resistance. Rhesus monkeys and ferrets are susceptible to P. mirabilis, making them useful as animal models for future studies on the mechanism of P. mirabilis-induced UTI and its corresponding treatment.

Methicillin-resistant Staphylococcus aureus (MRSA) continues to be a major cause of nosocomial infections. Methicillin resistance in S. aureus is caused by the acquisition of the mecA gene, located on a mobile genetic element called the staphylococcal cassette chromosome (SCC). The aim of this study was to evaluate the presence of the predominant SCCmec type present among clinical isolates. This cross-sectional study was performed on a total of 146 MRSA isolates obtained from clinical specimens between 2012 and 2013 from two major hospitals in Shiraz, Southwest of Iran. Antibiotic susceptibility profiles were determined by the disc diffusion method according to the guidelines of The Clinical and Laboratory Standards Institute. Bacterial DNA was extracted using the small-scale phenol-chloroform extraction method and was employed as polymerase chain reaction (PCR) templates for the assigned current SCCmec types. The assigned SCCmec types by PCR revealed the SCCmec type I as the predominant type with 86 (58.9%) samples, followed by the SCCmec type II with 29 (19.9%), type III with 16 (11.0%), and type IV with 12 (8.2%) samples, respectively. The SCCmec type I MRSA isolates were significantly recovered from blood (80%) and sputum (67.2%). The results of antibacterial susceptibility tests for the MRSA isolates showed that all of those carrying the SCCmec type I and II had significantly greater resistance rates to Gentamicin and Rifampin than the isolates containing the SCCmec type III. Also, a significant difference was detected for susceptibility to Co-trimoxazole between the SCCmec type I and II MRSA isolates and the SCCmec type III, which was more resistant. The frequency of the isolates containing type I in the current study can indicate an emergence of this SCCmec type in the studied medical centers.

Coagulase-negative staphylococci (CoNS) are opportunistic pathogens. Methicillin resistance is common in CoNS and may play an important role as reservoir of staphylococcal cassette chromosome mec (SCCmec) for Staphylococcus aureus. The aim of this study was to determine molecular characteristics of nasal carriage of methicillin-resistant coagulase negative staphylococci among students. MR-CoNS from both nares of students were collected. Resistance to methicillin was determined by cefoxitin (30μg) disk diffusion test. SCCmec typing was performed using multiplex PCR by mec complex classes and ccr genes. Antimicrobial susceptibility profiles were determined on Mueller-Hinton agar according to CLSI. A total of 600 consecutive students were enrolled in this study; 430 of whom (71.7%) had CoNS. Seventy-two MR-CoNS strains, 21 (29.2%) S. lugdunensis, 17 (23.6%) S. haemolyticus, 17 (23.6%) S. saprophyticus, 9 (12.5%) S. epidermidis and 8 (11.1%) S. schleiferi were isolated. MR-CoNS rate in nasal carriage was 16.7%. All strains were susceptible to vancomycin. Forty-eight (66.7%) had a single SCCmec type including types I (n = 5), II (n = 4), III (n = 7), IV (n = 19) and V (n = 13), whereas 5 (6.9%) had two types including III + IV (n = 2), III + V (n = 1) and IV + V (n = 2). Nineteen strains (26.4%) were non-typeable for their SCCmec and ccr. Types IV and V SCCmec were associated with S. lugdunensis and S. haemolyticus, respectively. SCCmec types IV and V were prevalent in MR-CoNS and few isolates could harbor more than one type.

Multiple sclerosis (MS) is a demyelinating condition affecting the central nervous system. Although the cause of this condition is unknown, patients with MS seem to have genetic vulnerability to certain environmental factors such as infection that could trigger this condition. We conducted this study to determine whether MS risk increases following primary infection with Epstein-Barr virus (EBV) and also to investigate any association between MS and seropositivity to anti-EBNA-1 IgG, anti-EBV-CA IgG, and anti-EBV-EA.Patients and EBV infection was confirmed using the Enzyme-Linked Immunoassay in the patient (n = 46) and control (n = 46) groups via commercial assays (anti-EBNA-1 IgG, anti-EBV-CA IgG, and anti-EBV-EA kits). The data were analyzed by using three statistical tests (Pearson chi-square, Spearman rho correlation, and odds ratio). Seropositivity to anti-EBNA-1 IgG did not show a significant difference between the patient and control groups (92.9% and 88.4%, respectively), and nor was seropositivity to anti-EBV-CA IgG different between the two groups (95.2% and 95.3%, consequently). The anti-EBV-EA-D test was negative in all the patients and in 95.3% of the controls. Seropositivity to both anti-EBNA-1 and anti-EBV-CA indicating past infection did not show significant associations with the later development of MS (Pearson chi-square asymptotic significance [Asymp. Sig.] [2-sided] = 0.317, Spearman's rho correlation test Sig. [2-sided] = 0.689, odds ratio = 1.95). Seropositivity to both EBNA1- IgG and EBV-CA- IgG did not show a causal association with MS. The findings of this study suggest that EBV past infection could not be a causative factor in the development of MS and a protective factor against classic MS.

The treatment of onychomycosis is a challenge and infections are typically more severe and difficult to treat in toenails than in fingernails. The current study aimed to investigate the fungicidal effect of ultraviolet radiation on the growth of dermatophytes isolated from nails.Patients and Samples from patients with clinical manifestations of onychomycosis were inoculated onto Sabouraud dextrose agar and incubated at 30°C for 14 days. Isolated species were identified by specific laboratory examinations; UV-A, UV-B, and UV-C light were used to irradiate two strains of Trichophyton mentagrophytes and T. rubrum. Colony count, size and growth rate of the isolated fungi were evaluated under laboratory conditions. Trichophyton rubrum type 1 was less sensitive to UV-A and UV-C, and more sensitive to UV-B than type 2. T. mentagrophytes type 2 was slightly responsive to UV-A therapy, although no decrease in colony count was observed. Increased doses of UV-B and UV-C irradiation decreased the counts. The effect of radiation on colony size was dependent on the dose and type of irradiation. UV-A, UV-B, and UV-C light seem to be effective in decreasing colony growth of the most prevalent fungi, which caused onychomycosis in the current study samples. Further studies are needed to determine the efficacy of ultraviolet light therapy, identify possible side effects, and establish appropriate dosages for the antifungal effect of this therapy.

Candida spp. is the most common organisms involved in fungal infections in the high risk patients. It causes the greatest number of invasive candidiasis. Fluconazole is effective in treating mucosal candidiasis. However, resistance to fluconazole and other azoles antifungal drugs is an important clinical problem to treat candidiasis. Caspofungin is more effective against Candida species such as some azoles-resistant isolates.. The current study aimed to investigate the susceptibilities of clinical fluconazole-resistant and fluconazole - susceptible dose- dependent Candida species to caspofungin. In the Minimum Inhibitory Concentration (MIC) test, 207 Candida species and other yeasts isolated from Iranian patients (each isolated from a high-risk patient) were evaluated. The yeasts were differentiated by standard mycological methods, CHROM agar Candida, and verified by API20C.AUX. In vitro susceptibilities were determined using Broth Micro Dilution (BMD) method described in the Clinical Laboratory Standards Institute M27-A3. MICs were noted after 24 and 48 hours of incubation. The most frequently isolated species were Candida albicans (52.2%), C. glabrata (24.6%), followed by C. tropicalis (7.7%) and C. krusei (3.4%). MICs of caspofungin against 87% of C. albicans and 90% of C. glabrata and C. tropicalis isolates were 2 μg/mL and for C. krusei were 4 μg/mL, respectively. The results revealed that only 20 out of 207 isolates (9.7%) were non-sensitive to caspofungin. Caspofungin non-susceptible isolates were isolated from the patients with cancer, diabetes and AIDS; and not in the species isolated from patients with other underlying diseases. Caspofungin appears more effective in vitro against Iranian fluconazole-resistant Candida isolates and some other yeasts.