Biosorption of copper, nickel, and manganese as well as the production of metal nanoparticles by Bacillus species isolated from soils contaminated with electronic wastes.

Improper electronic waste management in the world especially in developing countries such as Iran has resulted in environmental pollution. Copper, nickel, and manganese are from the most concerned soil contaminating heavy metals which found in many electronic devices that are not properly processed. The aim of this study was to investigate the biological removal of copper, nickel, and manganese by Bacillus species isolated from a landfill of electronic waste (Zainal Pass hills located in Isfahan, Iran) which is the and to produce nanoparticles from the studied metals by the isolated bacteria. The amounts of copper, nickel, and manganese in the soil was measured as 1.9 × 104 mg/kg, 0.011 × 104 mg/kg and 0.013 × 104 mg/kg, respectively based on ICP-OES analysis, which was significantly higher than normal (0.02 mg/kg, 0.05 mg/kg, and 2 mg/kg, respectively. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of metals on the bacterial isolates was determined. The biosorption of metals by the bacteria was evaluated by inductively coupled plasma optical emission spectroscopy (ICP-OES). The metal nanoparticles were synthetized utilizing the isolates in culture media containing the heavy metals with the concentrations to which the isolates had shown resistance. X ray diffraction (XRD), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDS) were used for the evaluation of the fabrication of the produced metal nanoparticles. Based on the findings of this study, a total of 15 bacterial isolates were obtained from the soil samples. The obtained MICs of copper, nickel, and manganese on the isolates were 40-300 mM, 4-10 mM, and 60-120 mM, respectively. The most resistant isolates to copper were FM1 and FM2 which were able to bio-remove 79.81% and 68.69% of the metal, respectively. FM4 and FM5 were respectively the most resistant isolate to nickel and manganese and were able to bio-remove 86.74% and 91.96% of the metals, respectively. FM1, FM2, FM4, and FM5 was molecularly identified as Bacillus cereus, Bacillus thuringiensis, Bacillus paramycoides, and Bacillus wiedmannii, respectively. The results of XRD, SEM and EDS showed conversion of the copper and manganese into spherical and oval nanoparticles with the approximate sizes of 20-40 nm. Due to the fact that the novel strains in this study showed high resistance to copper, nickel, and manganese and high adsorption of the metals, they can be used in the future, as suitable strains for the bio-removal of these metals from electronic and other industrial wastes.

Inactivation of Byssochlamys fulva during ohmic heating of tomato juice.

Ohmic heating is an emerging direct thermal technology, which uses electricity to heat food products volumetrically. Ohmic heating provides thermal and non-thermal effects like electropermeabilization to inactivate microorganisms. In this study, ohmic heating was used to inactivate Byssochlamys fulva in tomato juice. The main and interaction effects of initial pH (3.5 and 4.5) and voltage gradient (15 and 20 V/cm) were investigated on mold inactivation during ohmic heating at 88, 93, and 98 °C for 20, 10 and 5 min, respectively. The pH, acidity, total soluble solids, and Dvalue were compared. The results showed that pH and voltage gradient had significant effects on Dvalue and Zvalue (p < 0.05). In order to model the survival behavior of Byssochlamys fulva, due to the nonlinearity of the curves, Weibull model gave more accurate estimation compared to classical first-order model.

Bacteriophages PɸEn-CL and PɸEn-HO can eliminate MDR Enterobacter cloacae and Enterobacter hormaechei isolated from burn wound infections without toxicity for human skin cells.

The prevalence of multidrug-resistant (MDR) strains has caused serious problems in the treatment of burn infections. MDR Enterobactercloacae and Enterobacterhormaechei have been defined as the causative agents of nosocomial infections in burn patients. In this situation, examination of phages side effects on human cell lines before any investigation on human or animal that can provide beneficial information about the safety of isolated phages. The aim of this study was to isolate and identify the specific bacteriophages on MDR E. cloacae and E. hormaechei isolated from burn wounds and to analyze the efficacy, cell viability and cell cytotoxicity of phages on A-375 and HFSF-PI cell lines by MTT (3-(4, 5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide) colorimetric assay and lactate dehydrogenase (LDH) release assay. Phages were isolated from urban sewage Isfahan, Iran. Enterobactercloacae strain Iau-EC100 (GenBank accession number: MZ314381) and E. hormaechei strain Iau-EHO100 (GenBank accession number: MZ348826) were sensitive to the isolated phages. Transmission electron microscopy (TEM) results revealed that PɸEn-CL and PɸEn-HO that were described had the morphologies of Myovirus and Inovirus, respectively. Overall, MTT and LDH assays showed moderate to excellent correlation in the evaluation of cytotoxicity of isolated phages. The results of MTT and LDH assays showed that, phages PɸEn-CL and PɸEn-HO had no significant toxicity effect on A375 and HFSF-PI 3 cells. Phage PɸEn-HO had a better efficacy on the two tested cell lines than other phage. Our results indicated that, there were significant differences between the two cytotoxicity assays in phage treatment compared to control.

Efficacious antibacterial potency of novel bacteriophages against ESBL-producing Klebsiella pneumoniae isolated from burn wound infections.

BACKGROUND AND OBJECTIVES: Prevalence of extended spectrum ß-lactamase (ESBL) leads to the development of antibiotic resistance and mortality in burn patients. One of the alternative strategies for controlling ESBL bacterial infections is clinical trials of bacteriophage therapy. The aim of this study was to isolate and characterize specific bacteriophages against ESBL-producing Klebsiella pneumoniae in patients with burn ulcers. MATERIALS AND METHODS: Clinical samples were isolated from the hospitalized patient in burn medical centers, Iran. Biochemical screenings and 16S rRNA gene sequencing were determined. The phages were isolated from municipal sewerage treatment plants, Isfahan, Iran. TEM and FESEM, adsorption velocity, growth curve, host range, and the viability of the phage particles as well as proteomics and enzyme digestion patterns were examined. RESULTS: The results showed that Klebsiella pneumoniae Iaufa_lad2 (GenBank accession number: MW836954) was confirmed as an ESBL-producing strain using combined disk method. This bacterium showed significant sensitivity to three phages including PɸBw-Kp1, PɸBw-Kp2, and PɸBw-Kp3. Morphological characterization demonstrated that the phage PɸBw-Kp3 to the Siphoviridae family (lambda-like phages) and both phages PɸBw-Kp1 and ɸBw-Kp2 to the Podoviridae family (T1-like phages). The isolated bacteriophages had a large burst size, thermal and pH viability and efficient adsorption rate to the host cells. CONCLUSION: In present study, the efficacy of bacteriophages against ESBL pathogenic bacterium promises a remarkable achievement for phage therapy. It seems that, these isolated bacteriophages, in the form of phage cocktails, had a strong antibacterial impacts and a broad-spectrum strategy against ESBL-producing Klebsiella pneumoniae isolated from burn ulcers.

Isolation, characterization, and effectiveness of bacteriophage Pɸ-Bw-Ab against XDR Acinetobacter baumannii isolated from nosocomial burn wound infection.

OBJECTIVES: With emergence of drug resistance, novel approaches such as phage therapy for treatment of bacterial infections have received significant attention. The purpose of this study was to isolate and identify effective bacteriophages on extremely drug-resistant (XDR) bacteria isolated from burn wounds. MATERIALS AND METHODS: Pathogenic bacteria were isolated from hospitalized patient wounds in specialized burn hospitals in Iran, and their identification was performed based on biochemical testing and sequencing of the gene encoding 16S rRNA. Bacteriophages were isolated from municipal sewage, Isfahan, Iran. The phage morphology was observed by TEM. After detection of the host range, adsorption rate, and one-step growth curve, the phage proteomics pattern and restriction enzyme digestion pattern were analyzed. RESULTS: All isolates of bacteria were highly resistant to antibiotics. Among isolates, Acinetobacter baumannii strain IAU_FAL101 (GenBank accession number: MW845680), which was an XDR bacterium, showed significant sensitivity to phage Pɸ-Bw-Ab. TEM determined the phage belongs to Siphoviridae. They had double-stranded DNA. This phage showed the highest antibacterial effect at 15 °C and pH 7. Analysis of the restriction enzyme digestion pattern showed Pɸ-Bw-Ab phage was sensitive to most of the used enzymes and based on SDS-PAGE, protein profiles were revealed 43 to 90 kDa. CONCLUSION: Considering the potential ability of the isolated phage, it had an antibacterial impact on other used bacterial spp and also strong antibacterial effects on XDR A. baumannii. Also, it had long latency and low burst size. This phage can be a suitable candidate for phage therapy.

The protective role of Kombucha extract on the normal intestinal microflora, high-cholesterol diet caused hypercholesterolemia, and histological structures changes in New Zealand white rabbits.

OBJECTIVE: The aim of the current study was to investigate the effect of Kombucha extract (tea) on the normal intestinal microflora and histological structures in rabbits. MATERIALS AND METHODS: This study was a descriptive-analytical investigation. Thirty-two male New Zealand rabbits were randomly divided into 4 groups as follows: Normal diet (I), high-cholesterol diet (II), normal diet plus Kombucha extract (II), and high-cholesterol diet plus Kombucha extract (IV). Microbial cultures were taken from feces of rabbits before and after the applied treatments. The rabbits' blood was collected from the heart to determine the level of cholesterol, glucose and iron in the blood. Aorta and coronary heart microtome cut samples were prepared for detection of histological changes. RESULTS: Rabbit stool cultures before treatment with Kombucha extract included Enterobacter aerogenes, Providencia rettgeri, Proteus mirabilis, Pseudomonas aeruginosa and Klebsiella oxytoca. However, Escherichia coli, Enterobacter aerogenes, Pseudomonas aeruginosa, klebsiella pneumoniae and Hafnia alvei were found in stool cultures after treatment with Kombucha extract. Group IV had significantly lower blood cholesterol levels. Animals that received Kombucha extract only had lower fasting blood sugar (FBS) levels. Healthy rabbits that received Kombucha extract only and group (IV) showed a significant increase in iron (Fe) levels and a significant decrease in total iron binding capacity (TIBC) levels. In both groups III and IV, the right and left coronary arteries were completely normal and no lesions were observed in the intima. CONCLUSION: The results of this study showed minor changes in the intestinal microflora of rabbits after treatment with Kombucha extract and positive effects of this tea on some risk factors (hypercholesterolemia, arteriosclerosis, and FBS).

The Effect of Environmental Stresses on lipL32 Gene Expression in Pathogenic Leptospira spp. through Real-Time PCR.

Leptospirosis is a worldwide infectious and zoonotic disease. The incidence of this disease is high in temperate regions, especially in northern Iran. The aim of this study was to investigate the effects of temperature, pH, and Phyllanthus amarus plant extract on the lipL32 gene expression in pathogenic Leptospira spp. Fifty water samples were collected. Culture and PCR technique were used to isolate and identify the bacterium and the presence of the lipL32 gene. The samples were exposed to different temperatures and pH levels for one day and the Ph. amarus plant extract at different concentrations for one and seven days. RNA was extracted, and cDNA synthesis was performed for all the samples. All cDNAs were evaluated by the real-time PCR (SYBR green) technique. Out of the 50 samples, ten samples (20%), using PCR were determined to contain the pathogenic Leptospira. Fold change of the expression of the lipL32 gene associated with stresses was as follows: temperature stress of 40°C, 35°C, and 25°C reduced the lipL32 gene expression in all three isolates, especially in the isolates type 1. The pH stress, i.e., pH values equal to 8 or 9 reduced the gene expression in three types of isolates, and pH = 6 stress increases the lipL32 gene expression in the isolates of type 1. Ph. amarus plant extract stress reduced the mentioned gene expression only in isolates of type 2. Temperature and pH stresses could lead to differences in the expression level and cause the lipL32 gene expression decrease in three pathogenic isolates. The MIC results showed anti-leptospiral effect of Ph. amarus plant extract.

The role of the bla KPC gene in antimicrobial resistance of Klebsiella pneumoniae.

BACKGROUND AND OBJECTIVES: Klebsiella pneumoniae isolates that produce K. pneumoniae carbapenemase (KPC) have become a grave concern for the treatment of infections. KPC-producing strains are not only able to hydrolyze carbapenems but are also resistant to a variety of ß-lactam and non-ß-lactam antibiotics. The present study evaluated the prevalence of bla KPC in K. pneumoniae infections and determined the antimicrobial susceptibility of the isolates. MATERIALS AND METHODS: The K. pneumoniae isolates were identified by biochemical tests and confirmed by genotyping. The modified Hodge test (MHT) was performed to detect carbapenemases, and antimicrobial susceptibility was determined for all isolates by the disc diffusion method. Also, for MHT-positive isolates, supposed to carbapenemases isolates, broth microdilution method was used to measure the minimum inhibitory concentrations (MICs) of meropenem and colistin. RESULTS: The bla KPC genotypic evaluation revealed that only 5 of 96 isolates carried bla KPC genes. Antimicrobial pattern showed that isolates carrying bla KPC were resistant to cefepime, ticarcillin/tazobactam, and aztreonam discs. Also, results of broth microdilution method showed that KPC-producing K. pneumoniae was resistant to meropenem and colistin, according to the CLSI and EUCAST. CONCLUSION: In this study nearly half the isolates showed carbapenemase activity as shown by MHT results, but only few of them were carrying bla KPC. Thus bla KPC gene is not the main cause of resistance spread to carbapenems in Isfahan, Iran.

Evaluation of Modified Hodge Test as a Non-molecular Assay for Accurate Detection of KPC-producing Klebsiella pneumoniae.

Klebsiella pneumoniae carbapenemase (KPC) have become a major therapeutic challenge because of its increasingly fast dissemination throughout the world. Accurate detection of KPC is essential for optimal treatment. The Clinical and Laboratory Standards Institutes (CLSI) for fast detection of KPC producers currently recommend Modified Hodge Test (MHT) and Carba NP test. MHT can directly detect carbapenemase production in Enterobacteriaceae isolates. The current study was conducted to evaluate the capacity of MHT with two carbapenem disks for accurate detection of KPC. MHT was performed according to guidelines of CLSI to identify isolates with carbapenem resistance. In doing so, two substrates of MHT were assigned into two groups for examination: meropenem and ertapenem groups. A total of 96 non-repetitive clinical isolates of Klebsiella pneumoniae were tested. The presence of the bla KPC gene in each MHT-positive isolate was examined by PCR. A total of 54 isolates exhibited reduced susceptibility or resistance to carbapenems. Sensitivity of MHT with two carbapenem disks was similar. Specificity of the MHT with meropenem disk was 64% and with ertapenem disk was 53%. Detection of KPC by MHT with meropenem disk was found to be more effective than with ertapenem disk. Based on our results, the presence of KPC does not in itself influence the categorization of resistance. Therefore, the use of MHT with ertapenem disk for the rapid detection of KPC among K. pneumoniae for infection control should not be recommended.

An in vitro study of antileishmanial effect of Portulaca oleracea extract.

BACKGROUND & OBJECTIVES: Leishmaniasis is caused by protozoa of Leishmania genus and is considered as a zoonotic disease. It is a major public health problem worldwide, with high endemicity in developing countries like Iran. Various chemical drugs are used for leishmaniasis treatment, but their side-effects and the emergence of drug resistance have led to look for new effective compounds. The aim of this study was to introduce purslane (Portulaca oleracea) as a traditional and medicinal herb which might act as a valuable source for designing new pharmaceutical drug/lead against Leishmania sp. METHODS: This study was conducted in the laboratory of Seddigheh Tahereh Infectious Disease Research Center, Isfahan, Iran during the spring of 2015. The essence from the purslane plant was prepared through water distillation and the alcoholic extract was prepared through maceration method. The essence was dried, and diluted with DMSO (5%). Leishmania major promastigotes were cultured in 25 ΁ 2΀C temperature in the stationary phase of RPMI-1640 medium, enriched with 10% fetal calf serum and penicillin-streptomycin to yield higher quantity. The biological activity of herb essence was evaluated on L. major promastigotes and compared to glucantime reference drug using methylthiazole tetrazolium (MTT) colorometric assay. The optical density absorbance was measured with Eliza reader set, and the IC50 value was calculated at different time intervals. All tests were repeated thrice. Results were analyzed by using Tukey test and t-test. RESULTS: The IC50 values after 48 h, for glucantime against standard parasite promastigotes and clinical strains were equal to 12 and 19 mg/ml, respectively, whereas for purslane herb leaves and stems essence; it was equal to 360 and 680 mg/ml, respectively. Although, the glucantime pharmaceutical drug was more efficient compared to the investigated herb essence, the essense had significant effect on L. major promastigotes with increasing density (p <0.05). The ingredients of the herb leaves and stem essence were-Phytol, squalene, palmitic acid, ethyl- linoleate, ferulic acid, linolenic acid, scopoletin, linoleic acid, rhein, apigenin, and bergapten. INTERPRETATION & CONCLUSION: The study showed that essence of purslane has considerable antileishmanial effects and can stop the growth of parasites in the laboratory compared to glucantime. More experiments are necessary to investigate its effect on Leishmania parasite in animal model.

Analyzing pmrA and pmrB genes in Acinetobacter baumannii resistant to colistin in Shahid Rajai Shiraz, Iran Hospital by PCR: First report in Iran.

Acinetobacter baumanni is known as a worldwide emerging nosocomial infections and it is classified as one of the six dangerous microorganisms by Diseases Society of America. Multi drug-resistant strains of A. baumannii have been reported in recent decades, which may be a result of the high use of antimicrobial agents. Colistin is the last form of treatment against this organism. The presence of pmrA and pmrB genes in A. baumannii causes the resistance of this organism against Colistin. This cross-sectional study was performed on 100 samples of A. baumannii isolated from ulcer, urinary, respiratory, blood of patients admitted to the intensive care unit of Shahid Rajai Shiraz hospital within a 12-month period. The diagnosis was performed by microscopic and biochemical testing using microgen kits. Determining Colistin resistance was carried out by Diffusion Disc, Colistin antibiotic disc of MAST- England and E-test. The analysis of genes pmrA and pmrB genes was done by PCR. 100 A. baumannii samples were diagnosed out of which using diffusion disk 94 cases were sensitive to Colistin and 6 cases were resistant to it. The E-test results in resistant samples presented an MIC equal to 64 micrograms per milliliter. The PCR results in sensitive and resistant to Colistin samples presented the existence of pmrA and pmrB genes. The results indicated the presence of pmrA and pmrB genes that are the main reason of A. baumannii resistance against the last line of treatment of this organism to Colistin.

Antileishmanial Activity of Date (Phoenix dactylifera L) Fruit and Pit Extracts In Vitro.

Leishmaniasis is considered as a major public health problem worldwide. Current drugs in treatment of leishmaniasis have some limitations; thus, the current study was aimed to assess the methanolic extracts of pit and fruit of Phoenix dactylifera against Leishmania major promastigotes. L major promastigotes were cultured in RPMI 1640 and incubated at 25°C ± 1°C for 24, 48, and 72 hours. For obtaining the IC50 (half maximal inhibitory concentration) value, MTT assay was employed. Furthermore, promastigotes were examined in terms of morphology under light microscope. About 48 hours after treatment, IC50s were estimated 23 µg/mL and 500 mg/mL for methanolic extracts of pit and fruit of P dactylifera, respectively. Both extracts exhibited a dose and time-dependent antileishmanial activity against L major parasites. Also, some visible morphological changes were seen. This finding revealed both date fruit and pit, are effective against L major promastigotes. Further studies should be designed in future based on apoptosis induction in vitro and in vivo.

Atherosclerosis: process, indicators, risk factors and new hopes.

BACKGROUND: Atherosclerosis is the major cause of morbidities and mortalities worldwide. In this study we aimed to review the mechanism of atherosclerosis and its risk factors, focusing on new findings in atherosclerosis markers and its risk factors. Furthermore, the role of antioxidants and medicinal herbs in atherosclerosis and endothelial damage has been discussed and a list of important medicinal plants effective in the treatment and prevention of hyperlipidemia and atherosclerosis is presented. METHODS: The recently published papers about atherosclerosis pathogenesis and herbal medicines effective in the treatment and prevention of hyperlipidemia and atherosclerosis were searched. RESULTS: Inflammation has a crucial role in pathogenesis of atherosclerosis. The disease is accompanied by excessive fibrosis of the intima, fatty plaques formation, proliferation of smooth muscle cells, and migration of a group of cells such as monocytes, T cells, and platelets which are formed in response to inflammation. The oxidation of low density lipoprotein (LDL) to Ox-LDL indicates the first step of atherosclerosis in cardiovascular diseases. Malondialdehyde factor shows the level of lipoperoxidation and is a sign of increased oxidative pressure and cardiovascular diseases. In special pathological conditions such as severe hypercholesterolemia, peroxynitrite concentration increases and atherosclerosis and vascular damage are intensified. Medicinal plants have shown to be capable of interacting these or other pathogenesis factors to prevent atherosclerosis. CONCLUSIONS: The pathogenesis factors involved in atherosclerosis have recently been cleared and the discovery of these factors has brought about new hopes for better prevention and treatment of atherosclerosis.

The study of Nickel Resistant Bacteria (NiRB) isolated from wastewaters polluted with different industrial sources.

BACKGROUND: Pollution due to the heavy metals is a problem that may have negative consequences on the hydrosphere. One of the best procedures in removing the toxic metals from the environment is using metal resistant bacteria. RESULTS: In the present study eight nickel resistant bacteria were isolated from industrial wastewaters. Three of them were selected as the most resistant based on their Maximum tolerable concentration (8, 16 and 24 mM Ni2+). Their identification was done according to morphological, biochemical characteristics and 16SrDNA gene sequencing and they were identified as Cupriavidus sp ATHA3, Klebsiella oxytoca ATHA6 and Methylobacterium sp ATHA7. The accession numbers assigned to ATHA3, ATHA6 and ATHA7 strains are JX120152, JX196648 and JX457333 respectively. The Growth rate of the most resistant isolate, Klebsiella oxytoca strain ATHA6, in the presence of Ni2+ and the reduction in Ni2+ concentration was revealed that K oxytoca ATHA6 could decrease 83 mg/mL of nickel from the medium after 3 days. CONCLUSION: It can be concluded that the identified Ni resistant bacteria could be valuable for the bioremediation of Ni polluted waste water and sewage.

Effect of altitude on some blood factors and its stability after leaving the altitude.

The underlying mechanisms of altitude training are still a matter of controversial discussion. The aim of this study was to compare the hemoglobin concentration, red blood cell count and volume between normal and high altitude situations and their persistence after returning back from higher altitudes. The study population included male students of Ardal Branch, Islamic Azad University. Twelve apparently healthy individual with high level of physical activity, mean age of 22.6 ± 1.50 years were selected through purposive and available sampling method. In this study, blood samples were collected at different time and altitudes in order to compare the changes of Red Blood Cell (RBC), Mean Cell Hemoglobin (MCH), Mean Corpuscular Hemoglobin Concentration (MCHC) and Mean Cell Volume (MCV). The first blood sampling was conducted at the altitude of 1830 m. The subsequent blood samplings were conducted 48 and 72 h after reaching the altitude of 4000 m and 24, 48 and 72 h after returning back to the altitude of 1830 m. The statistical method used in this study was repeated measurement ANOVA. Red Blood Cell (RBC) changes between onset of climbing to 1830 m and 24, 48 and 2 h after sojourn at 1830 m height (after returning from 4000 m altitude) was significant. Mean Cell Hemoglobin (MCH) showed no significant change in any of the altitudes. MCHC changes between onset of moving toward altitude 1830 meters and 24, 48 and 72 h after sojourn at 1830 m height (after returning from 4000 m altitude) was also significant in addition, MCHC showed a significant difference between 24 h staying at 1830 m altitude with 48 and 72 h staying at 4000 m altitude. Mean Cell Volume (MCV) showed no significant difference between 48 and 72 h staying at 4000 m altitude and also between 24, 48 and 72 h staying at 1830 m altitude; however, there was a significant difference between onset of moving toward 1830 m altitude with 24, 48 and 72 h staying at 1830 m altitude and also 48 and 72 h staying at 4000 m altitude. The results showed that being in altitude has significant effect on RBC and MCHC.

Prevalence of the genital tract bacterial infections after vaginal reconstructive surgery.

Due to frequent childbirth, heavy lifting and the structure of the lives of rural women in Shahrekord region, Iran, cystocele and rectocele are of the main medical problems of the women in this area and for its correction, vaginal reconstructive surgery is needed which causes infection. The purpose of this study was to identify the bacteria causing infection after vaginal reconstructive surgery and performing antibiogram to help these patients for faster recovery. Patients enrolled this study were 92 who had undergone previous vaginal reconstructive surgery and now had infection. After examination, the group of patients taking antibiotics (n = 26) were excluded and the remaining 66 completed the study questionnaire. A gynecologist performed sampling; related tests (aerobic and anaerobic culture using an anaerobic culture gas pack jar and type A which provides absolute anaerobic conditions) were performed; antimicrobial susceptibility testing using Disk Diffusion Method was carried out; and the results were recorded. All the positive samples were polymicrobial. Gardnerella vaginalis in 20 cases (31%), peptostreptococci and anaerobic cocci in 9 cases (13.6%), staphylococcus aureus in 8 cases (9.1%), bacteroides and fusobacterium in 7 cases (10.6%), streptococcus group B in 4 cases (6%), yeast cells in 11 cases (16.6%) and Trichomonas vaginalis in wet mount of 4 (6%) existed. Anaerobic bacteria showed 85% sensitivity to clindamycin, 82% to chloramphenicol, 85% sensitivity to ceftizoxime and 45% to penicillin. Facultative anaerobic bacteria showed a sensitivity rate of 90% to ceftizoxime, chloramphenicol and cephalothin. According to our findings, the rate of vaginal bacterial infection in women with vaginal reconstructive surgery has increased; from which, infections with anaerobic bacteria origins have increased dramatically. We recommend antibiotic prophylaxis prior to genital reconstructive surgeries.

Assessing epidemiology of cutaneous leishmaniasis in Isfahan, Iran.

BACKGROUND & OBJECTIVES: Leishmaniasis has an annual incidence of 0.5-1.5 million new cases and is endemic in 88 countries throughout the world. About 90% of cases of cutaneous leishmaniasis (CL) are reported from seven countries including Iran. Evidence suggests the increased annual incidence of this disease in Iran. Intracellular protozoan parasite, Leishmania, is an obligatory parasite. Sandflies transfer infectious forms of the parasite or its metacyclic promastigotes to its vertebrate hosts such as humans by biting. In order to review the epidemiology of CL in Isfahan, Iran, factors such as incidence, disease causes, geographic features, age, and sex distribution, nationality, and occupation of patients, and the clinical spectrum of disease were evaluated. METHODS: During the study, 1315 patients with CL, who referred to the Dermatology and Leishmaniasis Research Center at Isfahan, were evaluated. RESULTS: The highest prevalence of CL was observed in fall (54%) and in northern areas of Isfahan (60.9%). Although CL was prevalent in both men and women, it had higher incidence in men (61.8%). The majority of patients (31.2%) aged 21-30 yr old. Most lesions were nodule-shaped (36.5%) and in upper extremities (48.3%) particularly in men (32.4%). While 81.2% of the subjects were Iranian, others were Afghani or with other nationalities. Most patients had multiple lesions on their bodies and 141 individuals (10.7%) had a previous history of disease. Among all occupations, the highest prevalence of CL was detected in students (18.1%). The response to treatment with compounds of meglumine antimoniate (glucantime) was better than other treatments. INTERPRETATION & CONCLUSION: Unfortunately, the results showed that the prevalence of CL has been increasing annually in some provinces of Iran, especially in Isfahan Province. Nevertheless, further studies are required to determine the vectors, reservoirs, and species of disease and to design appropriate strategies to control the disease.

Comparison of the effects of silver nanoparticles on pathogenic bacteria resistant to beta-lactam antibiotics (ESBLs) as a prokaryote model and Wistar rats as a eukaryote model.

BACKGROUND: Antimicrobial resistance in hospital pathogens is an important concern. It can cause longer hospital stays, increase costs, and contribute to increased mortality and morbidity in hospitalized patients. The aim of this study was to categorize and identify gram-negative bacilli capable of ESBLs production and to study the effect of MIC silver nanoparticles on bacteria strains and then study them in Wistar rats. MATERIAL AND METHODS: A total of 186 clinical samples in 3 hospital of Isfahan city was studied during 8 months. The ESBL assay was performed by disk diffusion method. Minimum inhibitory concentration (MIC) values were determined by agar dilution method. Additionally, ESBLs production was examined by using the standard ESBL disc and DDT (double disk approximation test) procedures. Student's T-test and ANOVA were used for statistical analysis of the data. The ESBL-producing bacteria were then subjected to minimum concentrations of silver nanoparticles and then examined in Wistar rats. RESULTS: Of the 186 patients studied, 140 (75.3%) had gram-negative bacilli containing ESBL and 46 (24.7%) had gram-negative bacilli without ESBL and the most prevalent bacteria was identified as Klebsiella pneumonia, with especially strong resistance to cefotaxime. All of these bacteria were sensitive to the silver nanoparticle solution with density of 100 ppm, but the 4 nm size did not show any significant difference from control group Wistar rats at 6 months. CONCLUSIONS: The results seem to indicate a direct correlation between silver nanoparticle solution concentration and the diameter of growth zone for ESBL-producing bacteria. Assays in our study were in vitro; if use of silver nanoparticle particles in vivo proves to be with adverse effects, it could be a valuable alternative to antibiotics.

Controllable synthesis of ZnO nanoparticles and their morphology-dependent antibacterial and optical properties.

ZnO materials with different morphologies have been synthesized via a simple solvothermal method using different solvents without any catalysts, templates or surfactants. The ZnO samples are employed in the inactivation of gram-negative Escherichia coli and gram-positive Staphylococcus aureus in MilliQ water. The photocatalytic activities of samples to degrade an azo dye, Acid Orange 74 (CI 18745), were also tested. XRD data showed that single-phase ZnO with the wurtzite crystal structure but different growth habits were obtained in the different solvents. SEM imaging illustrated that ZnO with flower-like, rod-like, and spherical shape were produced when water, 1-hexanol, and ethylene glycol were used as the solvent, respectively. The optical properties of the as-prepared ZnO materials were investigated by UV-vis absorption and photoluminescence spectra. The antibacterial efficiencies were affected by the physiological status of the bacterial cells, different morphologies and crystal growth habits, particle size and optical properties of ZnO samples. Results indicate that ZnO flower-like showed significantly higher photocatalytic inactivation than ZnO rod- and sphere-like against E. coli compared with S. aureus. It was found that the antibacterial activity of ZnO increased with decreasing crystallite size. The inactivation efficiencies for both organisms under light conditions were higher than under dark conditions. The obtained results were discussed according to the morphologies, optical and structural properties of ZnO powders as key parameters in photocatalytic and antibacterial activity.

Genetic analysis of clinical isolates of Leishmania major from Isfahan, Iran.

BACKGROUND & OBJECTIVES: Leishmaniasis is a geographically widespread severe disease which includes visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL). There are 350 million people at risk in over 80 countries. In the Old World, CL is usually caused by Leishmania major, L. tropica, and L. aetiopica complex of which 90% of cases occur in Afghanistan, Algeria, Iran, Iraq, Saudi Arabia, Syria, Brazil and Peru. Recently, Eslami et al (2011) reported a novel TRYP6 gene encoding tryparedoxin peroxidase from an Iranian L. major strain exhibiting homology with the related gene in a divergent genus of Kinetoplastida, the Crithidia. This prompted us to analyze the mentioned gene in 100 isolates obtained from patients with suspected CL. Consequently, we analyzed internal transcribed spacer 1 (ITS1) region, RNA polymerase II largest subunit (RPOIILS) and the mitochondrial DNA polymerase beta (DPOLB). METHODS: After obtaining samples from 100 patients, DNA extraction was performed and TRYP6 was analyzed using conventional PCR. All samples harbouring TRYP6 with smaller size (555 bp) were analysed based on three other regions: ITS1, RPOIILS and DPOLB genes. RESULTS: Results showed that 10% of the isolates have the same character as observed in our previous study. The ITS1-RFLP-PCR of this 10% isolates showed their similarity to the one from Crithidia fasciculata. RNA polymerase II largest subunit (RPOIILS) showed genetic diversity but the mitochondrial DNA polymerase beta (DPOLB) did not show any genetic diversity. CONCLUSION: This study might also help in solving the problems concerning Leishmaniasis outbreaks currently reported in Iran and some other endemic regions of the world.