Isolation of fecal Escherichia coli from fresh strawberries (fragaria ananassa) in Iran.

Background and Objectives: Microbial safety of the fresh fruits is a global concern. The production of strawberry (Fragaria ananassa) in Iran has grown 4.5% from 2016 to 2019. Little information is available about the microbiological quality of strawberry produced in Iran. The objective of this investigation was to assess the fecal Escherichia coli (FEC) contamination of fresh strawberries of south of Kerman province where is the centers of production and supply of strawberries to other parts of Iran. Materials and Methods: In this cross-sectional descriptive study, the FEC of a total number of 109 strawberry samples, which were collected from green-houses of strawberry of Kerman province during three months, were enumerated using the most probable number (MPN) as described by Iranian National Standards Organization, protocol No.2946, and interpreted using the latest statute released by Iran Veterinary Organization, Executive order number 2946, released in 2011. Results: MPN of FEC counted in strawberries ranged between <0.3 (n=37) and >110 (n=19) per gram (g) having a mean, mode, and median value of 250.3, <0.3, and 9.4 MPN/g, respectively. More than one-fourth of the samples (28.44%) were polluted with FEC at a level of >100 MPN/g. Conclusion: Our findings may be resulted from the sanitary quality of the farm and strawberries of the study area, which indicated that the microbial safety of the strawberries in this survey was not satisfactory, alarming public health.

Determination of frequency, multiple antibiotic resistance index and resistotype of Salmonella spp. in chicken meat collected from southeast of Iran.

BACKGROUND: Zoonotic food-borne pathogens such as Salmonella spp., which can be hosted by some raw foods, play a crucial role in ranking the public health of a country OBJECTIVES: The present study was conducted to assess the frequency, antibiotic resistance pattern and index of multiple antibiotic resistance (MAR) of Salmonella spp. in chicken meat METHODS: A cross-sectional survey was conducted from October 2017 to March 2018. One-hundred and fifty chicken meat samples were collected from meat stores in Zahedan, southeast of Iran and screened for contamination with Salmonella spp. using the polymerase chain reaction assay targeting the inv-A gene. Antimicrobial susceptibility testing was performed against 11 commonly prescribed antimicrobial agents in the veterinary treatment to calculate the MAR index RESULTS: The contamination rate was 2.7% (4/150). The antimicrobial resistance rate was 100% (n = 4) against penicillin, tylosin, tetracycline, erythromycin and tiamulin, 50% (n = 2) against trimethoprim/sulfamethoxazole, difloxacin and lincomycin/spectinomycin and 25% (n = 1) against flumequine and florfenicol. All isolates were sensitive to fosfomycin. Interestingly, all isolates (n = 4) exhibited different MAR patterns. Furthermore, the MAR index ranged from 0.45 to 0.81 CONCLUSIONS: In addition to the MAR index, which indicated that the isolate originated from a source where antibiotics were used to a great degree and/or in large amounts, the results showed that the chicken meat hosted resistant strains of Salmonella spp. in the study area. Overall, the findings indicated an important public health problem. To reduce this alarming signal, the poultry industry should implement control measures in the study area.

Association of the Interleukin-27 Gene Expression and Hepatitis B Virus Infection in Liver Transplanted Patients.

OBJECTIVES: Hepatitis B viral infection is among the most common causes of cirrhosis and hepatocellular carcinoma and a frequent viral indication for liver transplant. Cytokine-mediated immunity plays a critical role in introducing and promoting hepatitis B virus outcomes and in graft microenvironment. Interleukin 27 is a heterodimeric cytokine and a member of interleukin-6/interleukin-12 family. Interleukin-27 shows a broad range of pro- and antiinflammatory properties and plays a determining role during immune responses in combating hepatitis B virus. Therefore, in this study, the possible association between expressions of interleukin-27 gene with hepatitis B virus infection was evaluated in liver transplant patients. MATERIALS AND METHODS: In a cross-sectional study from liver transplant patients with the risk of hepatitis B virus infection who admitted to Namazi Hospital affiliated to Shiraz University of Medical Sciences, 50 patients were selected and subgrouped to 25 hepatitis B virus-infected and 25 noninfected ones between years 2011 and 2013. The 25 healthy controls also were enrolled in this study. The presence of hepatitis B virus infection was assessed using polymerase chain reaction and enzyme-linked immunosorbent assay protocols in liver transplant patients. In addition, the interleukin-27 gene expression level was analyzed using an in-house-SYBER Green real time polymerase chain reaction method. The rate of interleukin-27 gene expression level was statistically analyzed in studied patient groups and controls using the Livak (2-▵▵CT) method. RESULTS: The expression level of interleukin-27 gene was increased 10.27- and 2.36-fold in hepatitis B virus-infected and uninfected liver transplanted patients compared with healthy controls. CONCLUSION: Hepatitis B virus infection can lead to overexpression of interleukin-27 gene in liver transplant patients compared with uninfected ones and controls. However, further studies are needed to characterize the effective antihepatitis B virus effects of interleukin-27 in liver transplant patients.

Mutation Screening of Exons 7 and 13 of the TMC1 Gene in Autosomal Recessive Non-syndromic Hearing Loss (ARNSHL) in Iran.

BACKGROUND: Non-syndromic hearing loss (NSHL) is the most common birth defect and occurs in approximately 1/1,000 newborns. NSHL is a heterogeneous trait and can arise due to both genetic and environmental factors. Mutations of the transmembrane channel-like 1 (TMC1) gene cause non-syndromic deafness in humans and mice. OBJECTIVES: The aim of the present study was to investigate the association of TMC1 gene mutations of the locus DFNB7/11 in exons 7 and 13 in a cohort of 100 patients with hearing loss in Iran using polymerase chain reaction-single-stranded conformation polymorphism (PCR-SSCP), heteroduplex analysis (HA), and DNA sequencing. PATIENTS AND METHODS: In this experimental study, the blood samples of 100 NSHL patients were collected from 10 provinces in Iran. These patients had a mean age of 16.5 ± 2.01 years and 74.15% of their parents had consanguinity. DNA was extracted from specimens and mutations of exons 7 and 13 of the TMC1 gene were investigated using PCR-SSCP. All samples were checked via HA reaction and suspected specimens with shift bands were subjected to DNA sequencing for investigation of any gene variation. RESULTS: In this study, no mutation was found in the two exons of TMC1 gene. It was concluded from these results that mutations of the TMC1 gene's special exons 7 and 13 have a low contribution in patients and are not great of clinical importance in these Iranian provinces. CONCLUSIONS: More studies are needed to investigate the relationship between other parts of this gene with hearing loss in different populations through the country. More research could clarify the role of this gene and its relation with deafness and provide essential information for the prevention and management of auditory disorders caused by genetic factors in the Iranian population.

Detection of pap, sfa, afa, foc, and fim Adhesin-Encoding Operons in Uropathogenic Escherichia coli Isolates Collected From Patients With Urinary Tract Infection.

BACKGROUND: Uropathogenic Escherichia coli (UPEC) with its virulence factors is the most prevalent cause of urinary tract infection (UTI). OBJECTIVES: This study aimed to determine the occurrence of fim, pap, sfa, and afa genes among 100 UPEC isolates collected from patients diagnosed with UTI. MATERIALS AND METHODS: A total of 100 UPEC isolates were obtained from urine samples of patients with UTI. The prevalence of 5 virulence genes encoding type 1 fimbriae (fimH), pili associated with pyelonephritis (pap), S and F1C fimbriae (sfa and foc) and afimbrial adhesins (afa) were determined through PCR method. We also investigated the phylogenetic background of all isolates. In addition, the distribution of adhesin-encoding operons between the phylogroups was assessed. RESULTS: The prevalence of genes encoding for fimbrial adhesive systems was 95% for fim, 57% for pap, 16% for foc, and 81% for sfa. The operons encoding for afa afimbrial adhesins were identified in 12% of isolates. The various combinations of detected genes were designated as virulence patterns. The fim gene, which occurred in strains from all phylogenetic groups (A, B1, B2, and D) was evaluated and no significant differences were found among these groups. Conversely, significant differences were observed in relation to pap, afa, foc, and sfa operons. CONCLUSIONS: These results indicate that the PCR method is a powerful genotypic assay for the detection of adhesin-encoding operons. Thus, this assay can be recommended for clinical use to detect virulent urinary E. coli strains, as well as epidemiological studies.

The analysis of correlation between IL-12 gene expression and hepatitis B virus in the affected patients.

Hepatitis B viral (HBV) infection, which is one of the global public health concerns, is among the most common causes of cirrhosis and hepatocellular carcinoma. It was proposed that cytokine-mediated immunity plays a critical role in determining the outcomes of hepatitis B virus infection. Interleukin 12 (IL-12) is a heterodimeric cytokine that shows a broad range of immunoregulatory properties during immune responses and combats host invading pathogens. The main purpose of this study was to investigate the possible association between expression levels of IL-12 gene with HBV infection in patients with HBV infection. This clinical study was performed on 30 HBV patients and 30 healthy controls. SYBR Green Real-time PCR was performed to examine the expression level of IL-12 gene in HBV patients. Then, the rate of expression was calculated using the Livak ([Formula: see text] ) method. ΔCT of samples in the two groups were compared using t test method. PCR was also used for HBV-DNA evidence. The results of our study demonstrated that the difference in mean of IL-12 gene expression between healthy subjects and HBV patients is statistically significant.

Microbial Susceptibility and Plasmid Profiles of Methicillin-Resistant Staphylococcus aureus and Methicillin-Susceptible S. aureus.

BACKGROUND: Today, significant increase in the prevalence and emergence of methicillin-resistant Staphylococcus aureus (MRSA) is a serious public health concern and is likely to have a dramatic negative impact on many current medical practices. Therefore, identification of MRSA strains is important for both clinical and epidemiological implications. OBJECTIVES: The present study was carried out to determine the frequency of methicillin resistant; antibiotic susceptibility and plasmid profiles of S. aureus recovered from different types of clinical samples of patients in Zabol, Iran. MATERIAL AND METHODS: Clinical samples from 500 outpatient and hospitalized patients were tested for S. aureus. The susceptibility of 106 S. aureus to 11 antibiotics was evaluated by the disk diffusion method and Etest oxacillin strips. The presence of mecA gene was investigated by polymerase chain reaction (PCR). The plasmid profile patterns of all isolates were determined by a modified alkaline lysis method. RESULTS: A total of 67 (63.20%) strains were found to be MRSA isolates. Most of MRSA isolates showed high level of resistance to ampicillin, erythromycin, nalidixic acid, penicillin, and tetracycline. Twenty-six percent of MRSA isolates showed high level of resistance to oxacillin (minimum inhibitory concentration [MIC] ≥ 256 µg/mL). mecA gene was detected among 62 MRSA isolates. Totally, 75 isolates of both strains harbored plasmid. CONCLUSIONS: Resistance to oxacillin and other antibiotics was high, and most of the isolates were found to be multi-drug resistance (MDR). Plasmid analysis of representative S. aureus isolates also demonstrates the presence of a wide range of plasmid sizes, with no consistent relationship between plasmid profiles and resistance phenotypes. Regular surveillance of hospital infections and monitoring of their antibiotic sensitivity patterns are required to reduce MRSA prevalence. High prevalence and multi-drug resistance of MRSA isolates in southeast of Iran could be considered as an urgent warning for public health.

Cervico-vaginopathogenic Escherichia coli in Iran: Serogroup distributions, virulence factors and antimicrobial resistance properties.

Escherichia coli (E. coli) play an important role in urogenital infection in pregnant and non-pregnant women. They are classified based on various types of O antigen, virulence properties, phylogenetic background and antimicrobial susceptibility pattern. E. coli strains that cause genitourinary infections possess several genes encoding urogenito-virulent factors. The aim of the present study was to determine the prevalence of eight known urovirulence factors and its association with serotypes, phylogenetic background and antimicrobial susceptibility pattern. One hundred thirty two cervico-vaginal E. coli (CVEC) isolates from pregnant and non-pregnant women (18-55 years old) were surveyed for adhesins (fimH, iha), haemolysins (hlyA), outer membrane protease (ompT) and siderophore receptors (irp2, iroN and iucD), cytotoxic necrotizing factor 1 (cnf1), and 12 selected O serotypes by multiplex-PCR. E. coli isolates were classified into intraspecies phylogenetic groups by PCR amplifications of phylogenetic markers. Antimicrobial susceptibility was determined by Kirby-Bauer's disc diffusion method. The most frequently found virulence factor-encoding gene, in descending order were: fimH (71%), irp2 (63%), followed by ompT (45%), iucD (37%), and iroN (31%) genes. The most prevalent serogroups for all E. coli isolates were O25, O15 and O6. There was an acceptable correlation between serotype and genotype in CVEC. The most isolated strains belonged to the phylogenetic group B2, harboring all tested virulence genes. Resistance to ampicillin was most frequently observed, followed by resistance to amikacin and cefazolin. The results suggest that E. coli isolates from different infection origins may have different characteristics. A better understanding of these differences may lead to further development of evidence-based clinical guidelines for the management of cervico-vaginal infection.

Study of VSX1 mutations in patients with keratoconus in southwest Iran using PCR-single-strand conformation polymorphism/heteroduplex analysis and sequencing method.

OBJECTIVE: Keratoconus (KC) is an eye disorder in which the cornea is swollen, thinned and deformed. Despite extensive studies, the pathophysiological processes and genetic etiology of KC are unknown. The disease incidence is approximately 1 in 2,000, and it is the most common cause of corneal transplantation in the USA. Many genes are involved in the disease, but evidence suggests a major role for VSX1 in the etiology of KC. This study aimed to determine the frequency of mutations in exons 2, 3 and 4 of the VSX1 gene in Chaharmahal va Bakhtiari province in the southwest of Iran. STUDY DESIGN: In this experimental study, mutations in 3 exons, namely exons 2, 3 and 4, of VSX1 were investigated in 50 patients with KC and 50 healthy control subjects. DNA was extracted using a standard phenol-chloroform method. PCR-single-strand conformational polymorphism/heteroduplex analysis was performed, followed by DNA sequencing to confirm the identified motility shifts. RESULTS: H244R mutations were found in 1 patient and also in 1 healthy control subject. Furthermore, 12 polymorphisms were identified in patients with KC and 7 in healthy control subjects [rs6138482 and c.546A>G (rs12480307)]. CONCLUSION: Our investigation showed that KC-related VSX1 mutations were found in a very small proportion of the studied patients from Iran. Further investigations on other genes are needed to clarify their roles in KC pathogenesis.

The role of diclofenac sodium in the dimorphic transition in Candida albicans.

Diclofenac sodium is a non-steroidal anti-inflammatory drug that inhibits filamentation in Candida albicans. Here we examined the effect of diclofenac sodium on hypha formation in C. albicans. The C. albicans cells were treated with various concentrations of diclofenac sodium (50, 100, 200 and 500microg/ml) and incubated at 37 degrees C for 2h. The characteristics of hypha formation were then assessed microscopically in both liquid and solid media. The results indicated that the effect of diclofenac sodium was dependent on the concentration of this compound, and preincubation with 500microg/ml diclofenac sodium completely inhibited hypha formation in both liquid and solid media. RT-qPCR analysis of RNA extracted from C. albicans indicated that the levels of expression of agglutinin-like sequence 3 (ALS3), RAS1, EFG1 mRNA, which are regulated by the cAMP-EFG1 pathway in C. albicans and three hypha-specific genes (ALS1, ECE1 and HWP1), were decreased in diclofenac sodium treated cells compared to the levels in controls. Our results also demonstrated that diclofenac sodium possesses potent anti yeast-hypha transition activity in vitro and it could be useful in combined therapy with conventional antifungal agents in the management of treatment of Candida albicans infections.