Jundishapur Journal of Microbiology
Volume:7 Issue: 5, May 2014

Traditional medicine/complementary alternative medicine may suggest new ideas to modern medicine in order to face new challenges however these concepts should be acknowledged based on experimental studies. We aimed to study the effects of onion (Allium cepa) juice on the normal flora of conjunctiva and eye lids, and to follow the histopathology changes of conjunctiva in an animal study. Twenty-four rabbits were randomly classified into three equal groups. Groups 1, 2 and 3 received fresh red onion juice, as an eye drop, into the right eye twice daily for; one week, one month, and two months, respectively. Microbiological sampling by sterile swabs was performed before and after the intervention. Cultural characteristics, including the growth rate and the kind of organisms, are reported. At the end of the study, pathological samples were collected from the inferior fornix. After the intervention, the number of positive cultures in the samples, collected from both the conjunctiva and eyelid, had decreased. Group 3 demonstrated the lowest amount of growth after the administration of the onion juice and the bacterial isolation rates from each organism had decreased. All pathological samples revealed some degree of inflammation. There was no evidence of metaplasia or dysplasia. There was no significant difference between the growth rates of organisms in the experimental groups using statistical analysis. According to our experiment, onion has an inhibitory effect on the growth of normal eye flora; although the duration of onion juice instillation did not show any significant effect on the group results. Hence, this finding is an initiating point for further investigations into the antimicrobial properties of this herb to treat common eye infections, including conjunctivitis and blepharitis.

Many studies have been done on the epidemiology of Hepatitis E on general population, but the data among patients with end stage renal disease (ESRD) are few and give conflicting results. The aim of this study was to investigate the prevalence of hepatitis E virus (HEV) infection and its relationship in ESRD patients undergoing maintenance hemodialysis (HD).Patients and This cross-sectional study was carried out on ESRD patients treated with HD in Imam Khomeini Hospital, Ahvaz city, Southwest of Iran. Blood sampling of patients was collected immediately before the dialysis session and the serum were evaluated for anti-HEV IgG titers by enzyme-linked immunosorbent assays. The statistical package for social sciences (SPSS) version 15 software was used for data analysis. Out of 47 ESRD patients, 27 were male (57.4%) and 20 were female (42.6%), with mean age of 55.27 ± 8.1 years. The prevalence of anti-HEV antibody was 10.6 % (five patients, four male and one female). The mean age of HEV positive and negative patients were 58 ± 5.52 and 53.82 ± 15.55 years, respectively without any significant difference (P = 0.058). There also was no significant association between HEV and gender (P = 0.28). The mean time of HD in HEV positive and negative patients were 1224.2 and 1168.5 days, respectively with no significant association (P = 0.88). In addition, there also was no association between HEV and HCV (P = 0.61). According to the present study, the prevalence of anti-HEV IgG antibody was 10.63 % among chronic HD patients and there was no association between HEV, age, gender, duration of HD and HCV antibody titer.

Polyvinyl alcohol (PVA) is one of the well-known polymers, which has been used in numerous biomedical applications because of its good biocompatibility. Due to problems made by the therapeutics already used for leishmaniasis, the aim of this study was to evaluate the effect of PVA containing artemether in treating cutaneous leishmaniasis in BALB/c mice. Aqueous solution of PVA was prepared by mixing with Double Distilled Water. After preparation of PVA, 4.33 mg of each drug (main drug artemether and control drug 14% glucantime) was added to 100 g of prepared PVA-honey solution. The solution was incubated at 37°C and the release of artemether was evaluated by measuring absorbance at 260 nm wave length. In this study for treatment of mice lesion, we used PVA containing artemether and glucantime and this method was compared with ointment treatment. Mean diameters of lesions in mice treated with artemether were smaller than the control group and the differences were significant (P < 0.05). The mean lesion size of mice treated with PVA containing artemether in comparison with the group treated with ointment of artemether were smaller and the differences were significant (P < 0.05). PVA containing artemether is a new method for treatment of cutaneous leishmaniasis and according to the obtained results, artemether is an appropriate and effective drug, especially when used with PVA as a lesion dressing; thus we suggest that this method can be applied for the treatment of cutaneous leishmaniasis.

Genetic diversity has provided Plasmodium falciparum with the potential capacity of avoiding the immune response, and possibly supported the natural selection of drug or vaccine-resistant parasites. Merozoite surface protein-3 (MSP-3) has been used to develop vaccines and investigate the genetic diversity regarding P. falciparum malaria in Iran. The main goal of this study was to analyze the polymorphic antigen MSP-3 genes across southeast of Iran among four different districts, to identify the differences in the allele frequency and genetic diversity. Nested polymerase chain reaction amplification was used to determine polymorphisms of N-terminal region of the MSP-3 gene. A total of 85 microscopically positive P. falciparum infected individuals from southeast of Iran were included in this study. Of the 85 confirmed P. falciparum samples obtained from four different districts, 72 were successfully scored for MSP-3.The MSP-3 allele classes (K1 and 3D7 types) showed comparable prevalence in all districts. Overall frequencies of K1 and 3D7 allele classes were 94.5 % for both. Since no study has yet looked at the extent of P. falciparum MSP-3 in this geographic region, these data can be helpful to support development of a vaccine based on MSP-3 against malaria. There should be a comparative analysis in different seasonal peaks to indicate the allelic polymorphism of MSP-3 over a period.

Staphylococcus aureus, the major virulence factor of hospital and community acquired infections, secretes numerous exotoxins (super antigens), which may affect immunological and inflammatory status in psoriatic skin lesion. This study is designed to compare the S. aureus super antigens level in sera of psoriatic patients with normal cases (nevus).Patients and A case control study was performed in dermatology ward of Rasoul Hospital in Tehran, IR Iran (2008 - 2010). Staphylococcal super antigens (Entrotoxin A, B, D and TSST1) were measured in serum of 41 psoriatic patients and 28 normal persons (Nevus) by ELISA. Chi square values (CI 95%, P value < 0.05) were calculated for all categorical variables. In this study 63.4% (26) of cases were male, 36.6% (15) were female. Age ranged from 4 months to 64 years old, with a mean age of 33.7 ± 15.4 years. Type of skin disease in cases: 20% (8) were inflicted by the Gutate form of the disease; 59% (23) with chronic plaque psoriasis (CPP), 7.7% (3) with erythrodermic and 12.8% (5) had other types of the disease (plaque, pustular, inverse). TSST (toxic shock syndrome toxin) was detected in 47% (20/41) of cases and in 6% (1/28) of the controls with a significant difference. (P value = 0.000) Entrotoxins (A, B, D) were detected in the sera of 48.8% (21/41) of cases; and only 6 %(1/21) of controls, showed significant differences (P value = 0.000) positive TSST was more common in spring, and correlates with CPP type of psoriasis, but not related to patient`s gender and age. In this study, S. aureus were 25 times more in psoriatic patients. Super antigens should be first detected in the serum samples; if negative, the skin lesions should be examined by PCR especially in chronic types of disease. Adding the antibiotics against S. aureus to other conventional treatments might be helpful. It has a more important and significant role in children with acute infection.

Cervical cancer is the second most common cancer in women worldwide. Recent studies show that human papillomavirus (HPV) DNA is present in all cervical carcinomas and in some cervicitis cases, with some geographical variation in viral subtypes. Therefore determination of the presence of HPV in the general population of each region can help reveal the role of these viruses in tumors. This study aimed to estimate the frequency of infection with HPV in cervicitis, cervical adenocarcinoma, intraepithelial neoplasia and squamus cell carcinoma samples from the Isfahan Province, Iran.Patients and One hundred and twenty two formalin fixed paraffin embedded tissue samples of crevicitis cases and different cervix tumors including cervical intraepithelial neoplasia (CIN) (I, II, III), squamus cell carcinoma (SCC) and adenocarcinoma were collected from histopathological files of Al-Zahra Hospital in Isfahan. Data about histopathological changes were collected by reexamination of the hematoxylin and eosin stained sections. DNA was extracted and subjected to Nested PCR using consensus primers, MY09/MY11 and GP5+/GP6+, designed for amplification of a conserved region of the genome coding for L1 protein. In total 74.5% of the tested samples were positive for HPV. Amongst the tested tumors 8 out of 20 (40%) of CIN (I, II, III), 5 out of 21 (23.8%) of adenocarcinoma cases and 78 out of 79 chronic cervicitis cases were positive for HPV. The rate of different carcinomas and also the rate of HPV infection in each case were lower than other reports from different countries. This could be correlated with the social behavior of women in the area, where they mostly have only one partner throughout their life, and also the rate of smoking behavior of women in the studied population. On the other hand the rate of HPV infection in chronic cervicitis cases was much higher than cases reported by previous studies. This necessitates more attention to the role of human papillomaviruses in the their induction in the studied area.

The emergence of antimicrobial resistant strains of Escherichia coli has raised considerable interest in understanding the diversity and epidemiology of E. coli infections in humans. Virulence factors of E. coli determine the specific infections caused by this microorganism. This study aimed to determine the prevalence of eight E. coli virulence factors and their association with antimicrobial resistance in bacteria isolated from patients with urinary tract infections (UTI).Patients and One thousand patients with UTI were enrolled in this cross-sectional study. Antimicrobial susceptibility was examined by disc diffusion method according to CLSI guidelines. After DNA extraction, the materials were probed by PCR for eight virulence factors genes, namely fimH, hly, iucC, ibeA, sfa/foc, neuC, papC, and afa genes. The frequency of virulence factors papC, afa, sfa/foc, fimH, hly, neuC, ibeA, and iucC were 53.3%, 51.7%, 53.3%, 56.7%, 23.3%, 31.7%, 20%, and 73.3%, respectively. In addition, there was a high degree resistance to cotrimoxazole and nalidixic acid while a high degree of susceptibility to nitrofurantoin was detected. There was a statistically significant association between fimH gene and resistance to ciprofloxacin (P = 0.006), nalidixic acid (P = 0.025), and cotrimoxazole (P = 0.02). Such associations were found between ibeA gene and amikacin (P = 0.02) and cotrimoxazole (P = 0.02) as well as between afa gene and gentamycin (P = 0.05). The results showed that E. coli isolated from patients with UTI had eight virulence factors with high frequencies. Moreover, these results alleged a direct connection between virulence factors and antimicrobial resistance in E. coli.

Mycobacterium tuberculosis genotyping can effectively improve tuberculosis (TB) control programs by controlling disease transmission. Pulsed field gel electrophoresis (PFGE) is a particularly powerful tool for determination of clonal identity of bacteria providing information for understanding and controlling the spread of disease.

The aim of present study was to investigate the genetic diversity of M. tuberculosis strains in Khuzestan province by the PFGE technique.Patients and

In total, 80 M. tuberculosis positive cultures were obtained from tuberculosis patients. PFGE was performed on 60 PCR-confirmed isolates by using DraI and XbaI restriction enzymes according to standard protocols. Plugs containing digested DNA were then loaded on agarose gels and run using contour-clamped homogenous electric fields.

Fifty distinct DNA banding patterns were obtained by digestion of DNA with DraI and 38 DNA banding patterns by digestion with XbaI restriction enzymes. The patterns comprised of 17 different clusters in which cluster I was the major one, containing six strains. Three clusters contained three strains each and the 13 remaining clusters comprised of two strains each. Digestion with DraI yielded 15-20 DNA fragments with 50-485 kb size, while digestion by XbaI produced DNA fragments with a size smaller than 50-242 kb.

Despite the ability of PFGE for study of genetic diversity of many mycobacterial species and it being considered as a robust and useful tool, in this study we only found a 15% epidemiological relationship amongst the isolates. Thus, for higher discrimination of genotypic clusters among M. tuberculosis clinical isolates, the application of more sophisticated complementary techniques is required.

Acute toxoplasmosis may lead to congenital toxoplasmosis with fetal complications outcome during pregnancy. Anti- Toxoplasma gondii antibody seroprevalence is unclear in pregnant women of south of Khuzestan province, since limited data about T. gondii seroepidemiology has been published in pregnant women of this area (Abadan, Shadegan, Khoramshar). The aim of this study was to clarify the status of T. gondii seroprevalence in pregnant women of south of Khuzestan province. In this cross-sectional study, 501 full-term pregnant women were included. This study was carried out in Taleghani teaching hospital for six 6 months from May to October 2011. Informed consents signed by the patients were obtained. Blood IgG and IgM were measured using ELISA technique. The data was analyzed by SPSS 13 (Chicago, IL, USA). Chi-square test was used for comparison. The participants’ age range was 15 to 45 years (average: 27.4 ± 13). Of the 501 pregnant women, 70.65 % (n = 354) were seronegative for T. gondii IgG and IgM antibodies. There were statistical relationships between IgG seroprevalence and age, as well as IgG seroprevalence and cat holding. There was high percentage of seronegative (70.65 %) IgG and IgM antibodies in full-term pregnant women. They were susceptible to acute toxoplasmosis; thus, prenatal screening was recommend in our province after cost-beneficial analyses.

Staphylococcus aureus is one of the most common causes of nosocomial infections and its resistance to antibiotics is a global concern. Lysostaphin is an antimicrobial agent belonging to a major class of antimicrobial peptides and proteins known as the bacteriocins. It exhibits a high degree of anti-staphylococcal bacteriolytic activity. In this study, high level of recombinant mature lysostaphin in Escherichia coli was produced by using pET32a expression vector. The S. simulans gene encoding lysostaphin was extracted, amplified by polymerase chain reaction (PCR), and sub-cloned in prokaryotic expression vector pET32a. E. coli BL21 (DE3) plysS were transformed with pET32a-lys and gene expression was induced by IPTG. The expressed protein was purified by affinity-chromatography using (Ni-NTA) resin. PCR and sequencing results confirmed the successful cloning of the target gene into the vector. The expression of protein was induced by IPTG and high concentration of the recombinant protein was obtained via the purification process by affinity-chromatography.Our data showed that the recombinant mature lysostaphin protein produced by pET32a vector in E. coli system was very efficient.

There is a possibility for the presence of Helicobacter pylori in vegetables due to their close contact with polluted water, soil and feces.This study was carried out to detect the presence of H. pylori in vegetables and salads in Iran. In total, 460 vegetable and salad samples were collected and transferred immediately to the laboratory. All samples were cultured and tested for the presence of H. pylori using the Polymerase Chain Reaction technique. The results showed that 44 of 460 samples (9.56%) were positive for H. pylori using the culture method. The Polymerase Chain Reaction technique showed that 50 of 460 samples (10.86%) were positive for H. pylori. Un-washed leek, traditional salad, un-washed basil and un-washed lettuce were the most commonly contaminated samples. The presence of the bacteria in various vegetables was statistically significant (P < 0.05). Vegetables are a new source of H. pylori and accurate washing of vegetables improves such contaminations.

Infection with non-typhoid Salmonella (NTS) is one of the most important health problems all over the world. Antimicrobial drug resistance is increasing among Salmonella infantis species. The aim of this study was to investigate the frequency of presence of class 1 integrons in S. infantis species as well as its association with drug resistance. This cross-sectional study was performed on 50 S. infantis isolated strains, collected from chicken samples between 2009-2011. These strains were identified by standard biochemical tests and serology. Antibiotic susceptibility profiles and minimum inhibitory concentration determination for 14 antibacterial agents were performed using micro dilution and disk diffusion methods. The detection of class 1 integron was performed by the PCR method. The demographic and microbiological data for the integron positive and negative isolates were compared by SPSS software. Eighteen out of 50 (36%) of isolated S. infantis species had intl gene. The isolated bacteria were sensitive to cefotaxime and ciprofloxacin (100%). Also isolates were resistant to nalidixic acid, tetracycline and streptomycin. All isolate with class 1 integron were multidrug resistant. The result of this study showed that due to increased level of drug resistance in S. infantis and the presence of class 1 integron in these strains, resistance can be transferred to other food borne pathogens.

Biofilm formation of Staphylococcus epidermidis is a major etiological factor of inducing device-related infections. The ability of biofilm formation by the S. epidermidis was assessed in vitro on two brands of foldable (hydrophilic) and two brands of rigid (hydrophobic) intraocular lens materials in order to investigate the role of lens material in postoperative endophthalmitis. To ensure reproducibility of biofilm formation on intraocular lenses, two strains of S. epidermidis and three quantification methods were performed. The S. epidermidis strains, DSMZ3270 (biofilm-producer) and ATCC12228 (non-biofilm-producer) were applied. Organisms were cultivated on disks of different brands of foldable hydrophilic Intra Ocular Lens (IOL) made of acrylic (Didar, Iran; (A) and Omni, India; (B)), and rigid hydrophobic IOL made of polymethyl methacrylate (PMMA; Didar, Iran; (C) and Hexavision, France; (D)). Biofilms were stained with crystal violet (CV) dye, which is an index of biofilm formation. The bacterial population was counted after biofilm homogenization. Scanning electron microscopy (SEM) was performed to examine the extent of biofilm formation. Adherence of DSMZ3270 strain on both types of foldable and rigid IOLs, was significantly more than ATCC12228 (P < 0.001-0.05 and, P < 0.01-0.05, respectively). The bacterial populations between the lenses were significantly different (P < 0.05). Subsequent studies demonstrated significant differences between brands of foldable and PMMA IOLs. According to statistical analyses the incubation time influenced the biofilm formation on both types of IOLs which meant that by increasing incubation time, the biofilm formation increased. According to the SEM pictures, biofilm seems to be lysed at 72 hours. These data demonstrated that the attachment of bacteria to hydrophilic acrylic IOLs was more than hydrophobic PMMA ones independent of the brand. According to these results the bacterial strain might have more hydrophilic properties. Augmenting the biomass of biofilm by passing of time demonstrated the key role of time in biofilm formation on the IOL surfaces. The differences between IOL brands in the biofilm formation indicated the influence of design parameters for IOLs.

In a world of nanotechnology, the first concern is the potential environmental impact of nanoparticles. An efficient way to estimate nanotoxicity is to monitor the responses of bacteria exposed to these particles. The current study explored the antimicrobial properties of nZVI (zero-valent Iron nanoparticles) on the Gram-negative bacterial systems Erwinia amylovora, Xanthomonas oryzae and the Gram-positive bacterial systems Bacillus cereus and Streptomyces spp. The genotoxicity potential of nZVI was also assayed. The toxicity of nZVI was tested by two different Growing bacteria in liquid (broth dilution) and agar media (challenge test) containing different nZVI concentrations for 24-72 hours. The genotoxicity of nZVI was assessed using the preincubation version of the Ames test. The lowest concentrations of nZVI that inhibited the visible growth (MIC) of E. amylovora, X. oryzae, B. cereus and Streptomyces spp. were 625, 550, 1250 and 1280 ppm, respectively. The minimum bactericidal concentration (MBC) for E. amylovora and X. oryzae were 10,000 and 5,000 ppm of nZVI, respectively. MBC was not observed for the Gram positive bacteria. No bacteriostatic and bactericidal effects were observed for oxidized nZVI. Mutant frequency did not increase according to the vehicle control at the concentrations assayed, indicating a lack of mutagenicity associated with nZVI. nZVI nanoparticles are not mutagenic at low concentrations, therefore they can be used without detrimental effects on soil bacteria.

Diagnosis and control programs for infectious diseases among immigrants are the most important aspects of epidemiological studies for both origin and destination countries. Data about hepatitis B virus (HBV) infection among the Afghan immigrants in Iran is limited. To the best of HBV treatment and prevention in Afghan immigrants in Iran, the present study was conducted to determine the virus DNA level, and the frequency of respective hepatitis B risk factors among the respective seropositive patients in Fars province, southern Iran.Patients and A total of 64 HBsAg positive Afghan immigrants including 47 (73.4%) men and 17 (26.6%) women, with ages ranging between 15 and 74 years (mean ± standard deviation: 37.69 ± 15.02 years) participated in this study. From those, whole blood sample were collected and DNAs were extracted from the sera and analyzed by TaqMan real-time PCR assay with a set of primers and probe amplified core protein region of HBV genome. HBV DNA was detected in a total of 51/64 (79.7 %) serum samples; 37 (72.5%) male and 14 (27.5%) female. The copy number of HBV DNA ranged from 5 × 102 to 8.49 × 108 copies/mL in the serum samples; median 3.8 × 104 copies/mL. Demographic data and risk factors were also evaluated. The comparison of viral loads between the age groups and sex indicated no significant correlation (P > 0.05). However, the serum HBV DNA level significantly decreased in the treated patient group (P = 0.03). There was no significant difference in medicine usage between the two sexes in the study population (P > 0.05). Considering the results, determining the HBV DNA load and evaluation of treatment response can help to reduce the costs of diagnosis and treatment procedures in such patients, as well as, decreasing the risk of HBV transmission in immigrant Afghan population. Moreover, HBV screening strategies in country border entrances among immigrant should be performed. Moreover, free vaccination and treatment programs, and improving the level of HBV knowledge among Afghan immigrants in Iran is highly recommended.

In contrast with Escherichia coli, the association of E. hermanii with urinary tract infections has not been described. In this case, E. hermanii was the sole isolate recovered from urine specimens of a pyelonephritis patient. The organism was found to be susceptible to piperacillin-tazobactam, ceftazidime, cefazolin, cefixime, aztreonam, gentamicin, tobramycin, imipenem, meropenem and amikacin, and resistant to amoxicillin. Antibiotic treatment was initiated with oral cefixime (400 mg every 24 hours). The symptoms were relieved within 72 hours after therapy. A urine sample was taken seven days after antibiotic therapy. E. hermanii was no longer isolated. The present case demonstrates that the uropathogenic E. hermanii clone can cause destruction of the kidneys. During asymptomatic bacteriuria or cystitis, the bacteria remain in the urinary tract. Even when pyelonephritis develops, inflammatory response of the host is still restricted to the urinary tract. These signs mean that uropathogenic E. hermanii may be not very virulent.