Jundishapur Journal of Microbiology
Year:2014 | Volume:7 | Issue:6|Papers Count:17

Background: The protoscoleces of fertile hydatid cysts are considered as major risks in surgery and producing secondary cysts if rupture the cyst during operation and, cause infecting the dogs with adult worm if eaten by this animal. Bacterial infection of the hydatid fluid can lead to sterilization of the cyst.Objectives: The aim of this study was to determine the bacterial infection rate of livestock hydatid cysts in Hamedan, Iran, and test the isolated bacteria effects on viable protoscoleces, in vitro.Materials and Methods: A total of 5709 slaughtered livestock were inspected to detect the presence of hydatid cysts. The hydatid fluid of all cysts was cultured separately to isolate and identify the bacteria. The effect of isolated bacteria was tested on viable protoscoleces in culture tubes, in vitro. The culture tubes were observed and examined under a light microscope every two hours for 24 hours, and then, after 36 and 48 hours.Results: Infected cysts were found in 74% of animals in Hamedan (46% were calcified and the bacteria was isolated from 52%) and 62% in Borujerd. The isolated bacteria in the infected cysts were as follows: Escherichia coli, E. blattae, Klebsiella pnoumoniae, Proteus mirabilis, Enterobacter aerogenes, coagulase-positive and coagulase-negative Staphylococci, Pseudomonas aeruginosa and Edwardsiella tarda. The protoscoleces incubated with the isolated bacteria totally degenerated, but 55% of the protoscoleces in the control groups were intact and viable even after one week.Conclusions: This study indicated a high percentage of cysts bacterial infections in two provinces of Iran. The common isolated bacteria were E. coli and Klebsiella. The isolated bacteria degenerated the protoscoleces during short-time incubation, in vitro.

Background: Rotavirus (RV) is a major cause of gastroenteritis in infants and children and is one of the most severe public health problems. Rotaviruses outer layer contains two proteins including VP4 and VP7. These proteins are necessary for host-cell binding and penetration. TLP (triple layer virus particle) of RV is a complete infectious virion that binds to the target cells and internalized at the cytoplasm. The DLP (double layer virus particle) is a non-infectious particle that is formed through exclusion of the outer layer proteins including VP4 and VP7. These DLPs are the transcriptionally active forms of rotavirus.Objectives: The aim of this study was to transfer DLP of RV into cytoplasm of MA104 cells by Lipofectamine and to analyze their replication.Materials and Methods: Initially, rotavirus was purified by CsCl discontinuous gradient and DLP was separated from TLP based on density differences. For confirmation, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of the proteins were conducted Then the purified DLP of RV was transferred into MA104 cells using Lipofectamine.Results: We attempt to avoid the attachment and entry of the rotavirus by using Lipofectamine to mediate the delivery of viral particles directly into the cytoplasm. DLP was endocytosed into the cytoplasm following treatment by Lipofectamine and then replicated in cytoplasm.Conclusions: Therefore the non-infectious DLPs were became infectious if introduced into the cytoplasm of permissive and cancerous cells, without passing attachment and entry process.

Background: Determination of b-D-Glucan (BDG) in the serum aids to diagnose the invasive fungal infections. The current study evaluated the diagnostic potential value of BDG assay in monitoring the disease in experimental systemic candidiasis in a rat model. The results can provide a useful preliminary data to improve this approach in developing countries.Objectives: The present study aimed to evaluate b-D-Glucan assay in diagnosis and monitoring the systemic candidiasis in a rat model.Materials and Methods: Twenty one rats were infected with 106 Candida albicans blastospore per rat. Twelve rats were considered as the negative controls (six immunocompromised rats without infection and six intact rats). During a week, every 24 hours the BDG sera level was determined by both Fungitell and Wako kits. To confirm the systemic infection in each rat, the suspensions of their internal organs were cultivated on agar plates and the number of colony forming units (CFU) of C. albicans was counted.Results: All the infected rats were positive with BDG tests. An increasing level of BDG was observed during early days after injection. The cutoff value for discrimination of BDG positive sera was obtained from the negative sera by the Fungitell kit. The sensitivity, specificity, positive and negative predictive values assessed for the Fungitell kit were 95%, 66.6%, 90.47% and 80%, respectively. These criteria for those of Wako were 90%, 83.3%, 94.7% and 71.4%, respectively.Conclusions: While BDG assay seems to be a sensitive and specific adjunctive tool to diagnose and monitor the experimental systemic candidiasis, it seems that measuring the positive cutoff value in different laboratory conditions is necessary for favorable establishment of these tests.

Background: At present, limited clinical data is available regarding survival rates of patients co-infected with human immunodeficiency virus (HIV) /tuberculosis (TB) in developing countries.Objectives: The present study aimed to evaluate the effect of HIV infection on the survival chances of active TB adults who disclosed their symptoms of TB in this part of Iran.Patients and Methods: The records and data of 807 patients only infected with TB and 21 co-infected patients with HIV/TB, who were admitted to primary health care units in Iran, were evaluated. Their survival time was analyzed using the Kaplan-Meier Estimator, Log-rank test and SPSS version 16.Results: Cox regression analysis showed that co-infection with HIV significantly affects the survival rate of TB patients so that the rate of death was 20.7 (8.1-53) times more than TB infected patients alone. Also, married patients with tuberculosis were 2.7 times more at risk of death than single subjects. We also confirmed that in HIV/TB positive patients, married individuals were more prone to death than single subjects (P value<0.001).Conclusions: Our results denote the need to progress diagnostic and preventive measures in this part of Iran.

Background: The brown-banded cockroach Supella longipalpa (F.) as a mechanical vector of pathogens and source of allergens has recently become widespread in the city of Ahvaz, southwestern Iran.Objectives: This research was done to evaluate the efficacy of a dust-formulation of Metarhizium anisopliae isolate IRAN 437C, as a common entomopathogenous fungus, against S. longipalpa.Materials and Methods: Conidia dust-formulations of M. anisopliae were prepared in proportions of 1%, 5%, 10%, 25%, 50% and 100% with bad wheat flour as the carrier. Cockroaches were exposed to surfaces treated with 1.5 mg/cm2 of the formulations under laboratory and semi-field conditions.Results: Cockroach mortality rates increased and survival times (ST50) decreased with an increased proportion of conidia from 1% to 100% but records taken for mortality and survival time from proportions of 25%, 50% and 100% were not significantly different. The mortality rates reached 100% and 90-100% in adults and nymphs, respectively on the seventh day. The lowest ST50 was related to the proportion of 100% (3 days). Probit analysis indicated LD50 and LD90 values of 1.7×106 and 1.7×107 conidia/cm2 for adults and these values changed to 4.5×106 and 2.9 × 107 for third and fourth instar nymphs at three days post exposure. Proportion of 25% caused mortality rates of 87%, 81% and 73% in adult, adult & nymph and nymph populations, respectively at four days after exposure under room conditions.Conclusions: Conidia dust-formulation of M. anospliae isolate IRAN 437C could present a promising alternative to control the brown-banded cockroach.

Background: Cutaneous leishmaniasis is a health problem in the world. Lesions should be treated on cosmetically or functionally important sites, such as the face and hands. Cantharidin is a terpenoid compound produced naturally by beetles of Meloidae and Oedemeridae families.Objectives: The current study aimed to investigate the effect of cantharidin on Cutaneous Leishmaniasis (CL) lesions and IFN-ϒ and IL-4 patterns in infected BALB/c mice.Materials and Methods: Infected BALB/c mice were divided into five groups as: untreated (control group), eucerin-treated and 0.05%, 0.1% and 0.5% cantharidin-treated. Lesions diameter was measured by Vernier caliper every three days for four weeks. Cytokines levels were measured by enzyme-linked immunosorbent assay (ELISA) using U-CyTech kit.Results: The results indicated that treatment with cantharidin exacerbates lesions compared with the controls, except for 0.05% cantharidin dose that restrained lesion growth significantly. Interferon gamma level in cantharidin-treated groups was significantly less than that of the control group. But interlukin-4 level was similar among the groups.Conclusions: The current study results indicated that high doses of cantharidin exacerbates leishmaniasis lesion, but low dose of cantharidin inhibits lesion growth.

Background: Probiotic microorganisms are selected based on their long history of use as well as their lack of side effects. Nowadays, the consumption of probiotic products is growing intensively in developing countries. Researchers who work in the food industry and research centers pay more attention to the identification of new probiotic bacteria with better performance characteristics as well as investigation of their performance because these findings can be very effective in promoting sale and consumption of these products.Objectives: Hence, this study was performed following these objectives: isolating indigenous lactic acid bacteria from traditional dairy products in Markazi Province, screening strains with probiotic characteristics, identifying strains and performing microbial collection of probiotic strains with indigenous potential.Materials and Methods: In this study, the samples were screened from traditional dairy products, such as fresh yogurt, curd, Tarhana and Ghareghoroot of Markazi Province villages. Samples were enriched in de Man, Rogosa and Sharpe (MRS) broth, and different strains were isolated and purified from this culture on MRS agar medium. Isolated strains were investigated by microscopic observations, considering the following factors: catalase capability, resistance to acid and bile, bile salt hydrolysis and antibiotic susceptibility pattern.Results: Nineteen Gram-positive and catalase-negative strains belonging to the Lactobacillus genus were isolated from the above-mentioned diary samples. Seven strains were resistant to acid and bile in which acid resistance was between 21.08% and 122.33% and bile resistance was between 94.08% and 152.93%, respectively. All isolated strains were susceptible to different antibiotics and a small percentage had the ability to hydrolyze Sodium Taurocholate.Conclusions: There are many of different species of Lactobacillus probiotics in traditional dairy products of the Markazi province, based on the findings of this study. It is recommended for researchers to isolate these strains and investigate their probiotic characteristics in order to reproduce them for use in food production as well as for medical treatment.

Background: The lesions induced by Helicobacter pylori in a candidate animal model should always be examined thoroughly. The resemblance of these lesions to those observed in humans can indicate whether the usage of this model will contribute to the understanding of the various pathogenic mechanisms involved in the development of human H. pylori-associated diseases.Objectives: The aim of this study was to perform a histopathological and bacteriological evaluation of gastric lesions based on H. pylori and Helicobacter-like organisms (HLOs) in cats.Materials and Methods: The present study was carried out on 28 cat’s (13 male and 15 female cases) gastric mucosae, which were tested by bacteriological and histopathological methods. Biochemical tests such as catalase, oxidase and urease were utilized in addition to Gram and Giemsa staining.Results: This research demonstrated that solely one case of H. pylori was isolated by gastric mucosal culture. Microscopically, the infected stomachs by HLOs comprised a mild to severe diffuse lymphoplasmacytic infiltration into the subglandular and gastric mucosa. Lymphoid follicles were also marked, particularly within pyloric tissues and mostly in displaced mucosal glands. For 75% of the gastritis cases, both HLOs and rapid urease tests were positive, whereas 83% of cases were more than one-year-old with gastritis. Furthermore, 75% of cats indicated gastritis, though 25% encompassed no gastritis; hence 20% had negative results for the rapid urease test and 25% for the Giemsa staining test. Such results may indicate that cats without gastritis were considered as free of HLOs pathogenic bacteria.Conclusions: These results suggest that most cases of gastritis were located in the antral region. Additionally, the isolation of H. pylori from domestic cats raises the possibility of zoonotic characteristics for the slightly pathogen; therefore transmission occurs from cats to human and vice versa.

Background: Current diagnosis of Helicobacter pylori infection by biopsy-based tests requires invasive sampling. Non-invasive methods such as the H. pylori stool-antigen (HpSA) test may be the best alternative for diagnosis of active infection. However, due to the presence of antigenic-diversity among the strains, various commercial tests have shown some discrepancies in different geographical-areas.Objectives: This study evaluates a homemade HpSA kit developed by using the H. pylori antigens from Iranian-isolates for detection of H. pylori in the stool of infected patients.Patients and Methods: Based on the endoscopic features and/or a rapid-urease test (RUT), 30 child and 50 adult patients, were recruited. From these candidates, three biopsies for RUT, culture and histology, and a stool-sample, were obtained. Patients were considered as H. pylori-positive if culture alone or RUT plus histology were found to be positive. Presence of H. pylori antigens in their stools was detected by the homemade HpSA test and an imported HpSA kit (Immundiagnostik, Germany).Results: Using the biopsy-based tests with RUT, histology and culture, 53% (16.30) of children were diagnosed as H. pylori–positive while using the imported kit 57% and the homemade kit 50% of the candidates showed positive results. Also by the biopsy-based tests, 54% of the adults were diagnosed as H. pylori-positive while by the homemade kit 56% showed positive results. Considering the biopsy-based tests as the gold standard, sensitivity and specificity for the imported kit was 94% and 86%, respectively, while the mean sensitivity and specificity for the homemade kit was 96% and 98%, respectively.Conclusions: The homemade kit, compared with the imported kit and biopsy-proven tests may be a valid and reliable method for determining the presence of H. pylori infection in Iran.

Background: Tatumella ptyseos is a rod-shaped, Gram-negative, facultative, and anaerobic bacteria categorized in the Enterobacteriaceae family. It is a rare food-borne opportunistic pathogen which causes neonatal sepsis, bacteremia, and urinary tract infections. T. ptyseos has been also cultured from various food sources around the world.Objectives: It is difficult to determine the source of the infection in the patients (especially newborns) due to low information about the epidemiology of T. ptyseos. The current study aimed to investigate the isolation, identification and antimicrobial susceptibility pattern of T. ptyseos strains from the consumed powdered infant formula milk (PIF) in hospital neonatal intensive care unit (NICU).Materials and Methods: A total of 125 powdered infant formula milk (PIF) samples were purchased from drug stores from June 2011 to March 2012. T. ptyseos was isolated according to food and drug administration (FDA) method. For final confirmation, biochemical testes embedded in API-20E system were used. Drug susceptibility test was performed using the disc diffusion method, according to clinical and laboratory standard institute (CLSI) recommendations.Results: Results of the study showed that, out of 125 samples, T. ptyseos was isolated from four (3/2%) PIF samples. All isolated strains (100%) were resistant to ampicillin, carbenicillin, cotrimoxazole and amoxicillin.Conclusions: The present study was the first report on the isolation and identification of T. ptyseos from PIF in Iran. T. ptyseos are frequently present in various kinds of foods; therefore, further investigation on these samples is required. It is necessary to track the T. ptyseos in a wide variety of foods and individuals especially in immunocompromised people such as human immunodeficiency virus (HIV) -positive patients to reveal the possible routes of transmission of this pathogen to humans. In addition, molecular studies are required to determine the genetic relationship between T. ptyseos strains isolated from different sources.

Background: Protein A is the virulence factors of Staphylococcus aureus rolling in its pathogenesis, and its gene is used for typing. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) with different enzymes has been used for this action.Objectives: In this study, we used Bsp143I enzyme for digestion of the gene, coding protein A (spa gene) in S. aureus. The bacteria were isolated from patients and healthy carriers in Gorgan, north of Iran.Patients and Methods: DNAs of 128 S. aureus subjects (53 from healthy carriers and 75 from patients) were extracted and amplified using specific primers of the spa gene. The product was digested by Bsp143I enzyme and its pattern was assessed by gel electrophoresis.Results: There were seven spa types among the tested S. aureus samples, among which six types differed in the repeated X region of the spa gene, but the seventh type had a deletion on one of BSP143I restriction sites. The frequency of spa types among isolated S. aureus samples as well as healthy carriers was six and five, respectively. S. aureus isolated from wounds showed the most diverse spa types (five) among clinical samples. Types 1, 2 and 4 were observed in all clinical samples, while only one case of type 3 was identified among patients, whereas this type constituted over 32% of the isolates among carriers. We found seven and four spa types among methicillin-resistant S. aureus (MRSA) and methicillin-sensitive S. aureus (MSSA) isolates, respectively.Conclusions: Our results showed that typing the spa gene using PCR-RFLP using Bsp143I was an acceptable method for typing S. aureus. Furthermore, this survey showed that the types in healthy carriers and MSSA were more variable than patient and MRSA isolates, respectively. We used the Bsp143I enzyme, which was not used in any previous studies on the spa gene. The results of this study suggested that we can use PCR-RFLP of spa gene by Bsp143I for molecular typing and sequencing of S. aureus, instead of relatively expensive methods. This method is relatively rapid and inexpensive, and can be accomplished in centers with conventional molecular facilities.

Background: L-tryptophan is an important ingredient in medicines, especially in neuromedicines such as antidepressants. Many commercial processes employ various microorganisms with high tryptophan synthase activity to produce L-tryptophan from indole and L-serine, but these processes are very costly due to the costs of precursors, especially L-serine.Objectives: For this reason, we studied the ability to use processed Iranian cane and beet molasses as L-serine sources for L-tryptophan production, which enables us to reach a cost-effective process.Materials and Methods: Whole cells of Escherichia coli ATCC 11303 were induced for L-tryptophan synthase by addition of indole to the growth medium and bacterial cells harvested from the growth medium were used as biocatalysts in the production medium. Conditions of the production medium were optimized and Iranian cane and beet molasses were processed by solvent extraction with ethanol and n-butanol and used as L-serine sources of the production medium. Amount of L-tryptophan and theoretical yield of L-tryptophan production were determined by High Performance Liquid Chromatography and by a colorimetrical method on the basis of the remaining indole assay, respectively.Results: L-tryptophan production increased by 15 folds, when indole was used as an inducer. L-tryptophan was produced from processed Iranian beet molasses in satisfactory amounts (0.53 mM) and no exogenous pyridoxal phosphate was required as a cofactor under our experimental conditions.Conclusions: The obtained results proved that Iranian beet molasses include significant amounts of L-serine that makes them a suitable substitution for L-serine. Findings of the present study give impetus to use of Iranian beet molasses for cost-effective L-Trp production in the amino acid industry.

Background: Whooping cough was considered as one of the major causes of childhood morbidity and mortality worldwide. Resistant isolates of Bordetella pertussis to macrolides in some countries have been recently reported.Objectives: Recent reports on macrolide-resistant B. pertussis isolates and lack of evidence for such resistance in clinical isolates of the Iranian patients led the authors of the current study to study antibiotic susceptibility of the collected isolates in the country. Susceptibility of the B. pertussis isolates to three antibiotics was studied. Relatedness of the strains recovered in this research was also examined.Materials and Methods: The antibacterial activities of erythromycin, azithromycin, and clarithromycin antibiotics against the recovered isolates of 779 nasopharyngeal swabs were examined using MIC (Minimum Inhibitory Concentration) method. Relationship of the strains was characterized by Pulsed-field Gel Electrophoresis (PFGE).Results: Among the specimens, 11 cases (1.4%) were culture-positive. Among these isolates, only two isolates had high MIC values for erythromycin and clarithromycin. Pulsed-field gel electrophoresis analysis of the isolates revealed 6 PFGE profiles (A-F) among which three and two isolates had the same patterns in profiles A and B, respectively.Conclusions: Azithromycin can be a good drug of choice to treat patients infected by B. pertussis in Iran. Clonal relationship of the isolates showed that the same B. pertussis strains were isolated from different patients in Iran.

Background: Toxoplasma gondii is a zoonotic parasite, which is assumed to have cosmopolitan distribution.Objectives: Adopting a cross-sectional study design the current research aimed to determine the occurrence of the parasite in cattle, camels and sheep in Isfahan and Chaharmahal va Bakhtiary provinces of Iran.Materials and Methods: Animals in the field and those brought for slaughter at abattoirs were included. Blood samples were randomly collected from animals and investigated by polymerase chain reaction (PCR).Results: T. gondii infections were detected in 0.00%, 6.60% and 17.9% of the sample cattle (n=155), camels (n=122) and sheep (n=95) respectively. Sheep were more frequently affected in Chaharmahal va Bakhtiary (33.33%) compared to Isfahan (8.47%) (P=0.005, 95%; CI=6.88-43.35). No statistically significant difference was observed in infection prevalence between camels and sheep; and between the different sex categories in both camels and sheep.Conclusions: Evidence of T. gondii occurrence in sheep and camels was provided in the provinces under study. There is a need to investigate the potential risk factors of zoonotic infections. Furthermore, animal health and production losses caused by the parasite; and associated zoonotic implications in the area under study need to be explored.

Background: Extended-spectrum b-lactamase (ESBL) production is the major resistance mechanism to b-lactam antibiotics in Enterobacteriaceae. In addition, emergence of plasmid-mediated quinolone resistance (PMQR) in ESBL-producing isolates has become a global threat for treatment of these infections.Objectives: We investigated the association between ESBL production and quinolone resistance in urinary isolates of K. pneumoniae.Patients and Methods: A total of 196 urinary isolates of K. pneumoniae were collected from Imam Hussein Hospital in Tehran during a four year period (2008-2012). Antibiotic susceptibility was determined by disc diffusion and ESBL production was screened using the phenotypic confirmatory test (PCT).Results: All isolates were susceptible to imipenem. Resistance to piperacillin and cefotaxime were 66.3% and 50.5%, respectively. Resistance to ceftazidime, amoxiclave, aztreonam, ceftriaxone, cefepime, nitrofurantoin, gentamicin, ciprofloxacin, nalidixic acid, ofloxacin, norfloxacin, levofloxacin, amikacin and pipracilin/tazobactam were less than 50%. ESBL production was detected in 92 isolates (46.9%) of which, 61.9% were resistant to nalidixic acid and 65.2% to ciprofloxacin. Multidrug-resistance was observed in 96.7% of ESBL producers.Conclusions: Our results showed coexistence of ESBL and quinolone resistance in the majority of the uropathogenic K. pneumoniae test isolates suggesting that care should be taken for the choice of antibiotic therapy.

Background: Streptomyces avermitilis, belonging to Actinomycetes, is specialized for production of avermectin, used as an anthelmintic and insecticidal agent. It is mostly found in soil and its isolation is very crucial for medically important avermectin production.Objectives: In the present study, 10 bacterial isolates lacking antimicrobial activities were isolated from the soil samples collected from different areas of Lahore, Pakistan.Materials and Methods: Three distinctive localities of Lahore were opted for soil assortment to isolate S. avermitilis. About 50 isolates of Streptomyces species were attained through selective prescreening procedures. All of these isolates were studied for production of the secondary metabolite, avermectin. Different test like soluble pigment color and melanin formation were used for identification. Biochemical characterizations of those isolates closely resembling the control in morphological characteristics, soluble pigment color and melanin formation tests were performed.Results: The 10 selected isolates were identified as the avermectin-producing strain by fermentation and characterized on ISP2 medium for aerial and reverse side mycelia color, soluble pigment color and melanin formation, in comparison with S. avermitilis DSM 41445. The best avermectin-producing isolate S1-C (10.15 mg/L) showed similar result as S. avermitilis DSM 41445, when subjected for culture characteristics analysis in different media along with biochemical characterization.Conclusions: From the results, it was concluded that agricultural lands around Pakistan Council of Scientific and Industrial Research (PCSIR) Campus Lahore were rich sources of industrially important Streptomyces, especially S. avermitilis.

Background: Occult hepatitis B infected (OBI) patients cannot eradicate hepatitis B virus (HBV) -DNA from their liver and peripheral blood, completely.Objectives: The main aim of this study was to investigate the rate of HLA-A2 expression on peripheral blood mononuclear cells (PBMCs) of patients with OBI.Materials and Methods: In this experimental study, intensity of HLA-A2 was measured on the PBMCs of 57 OBI patients and 100 HBsAg-/anti-HBc+/HBV-DNA samples were enrolled as controls; measurements were performed using the flow cytometry technique.Results: Flow cytometric analysis indicated that 19 (33.3%) OBI patients and 28 (28%) controls expressed HLA-A2 antigen on their PBMCs. There was no significant difference between the two groups regarding the rate of individuals expressing HLA-A2 antigen. Statistical analyses showed that the intensity of HLA-A2 expression significantly decreased in OBI patients (3.58±0.1) in comparison to healthy controls (4.21±0.25; P<0.001).Conclusions: According to these results it can be concluded that decreased intensity of HLA-A2 on the PBMCs of OBI patients may lead to resistance of HBV in the patients.