International Journal of Molecular and Clinical Microbiology
Volume:7 Issue: 1, Winter and Spring 2017

We examined 170 formalin-fixed paraffin-embedded samples from esophageal squamous cell carcinoma patients. All subjects live in Mazandaran province, a region with high incidence rate of esophageal cancer and have become known as the “Asian Esophageal Cancer Belt”. Samples were tested for HPV-DNA by MY09/11 and Gp5ﾯ general primers using nested PCR. Of the 170 ESCC samples, 86 (50.6%) were male and 84 (49.4%) were female. The mean age of the subjects was 66.5±11.1 and ranged from 35 to 91 years. Totally, HPV-DNA was detected in 62 (36.5%) of the esophageal squamous cell carcinoma samples by HPV L1 consensus primers. Considering the location of esophagus specimens, of 62 positive samples, 16 (25.8%) samples were in the upper third, 28 (45.2%) in the middle third, and 18(29.0%) in the lower third. The current study showed a relatively substantial prevalence of HPV infection in esophageal squamous cell carcinoma samples in Mazandaran province.

The aim of this study was to investigate the prevalence of virulence and Cytolethal Distending Toxin (CDT) genes in the Campylobacter isolates from intestinal contents and gall bladders of broilers and, to evalute their cytotoxic effects on HeLa cell cultures. These genes play important roles in bacterial adherence to intestinal mucosa, flagella-mediated motility, invasive capability and the ability to produce toxins in Campylobacter pathogenesis. A total of 121 Campylobacter isolates (106 C. jejuni, 11 C. coli, 2 C. lanienae, and 2 C. lari) were used in this study. The frequency of virulence genes in all the isolates were detected in different proportions ranging from 34-93% using Polymerase Chain Reaction (PCR) assay. Cytolethal Distending Toxin A (CDTA), Cytolethal Distending Toxin B (CDTB) and Cytolethal Distending Toxin C (CDTC) genes were found in 66.1%, 65.3% and 66.9% of the Campylobacter isolates tested, respectively (P> 0.05). Of the 19 isolates, only two (one C. jejuni, one C. coli) showed morphological changes such as cell swelling, expansion, growth, and cell shape change in HeLa cell cultures. CDT and virulence genes were detected at low frequencies in C. jejuni, C. coli and C. lari isolates that were obtained from clinically healthy broilers.
Although valuable information was attained about the pathogenicity of C. lanienae, additional studies using animal models are necessary for clarification.

Background/ Aflatoxin M1 (AFM1) is a metabolite of the aflatoxin B1 found in the liver of livestock, as a result of feeding livestock with contaminated food. Certain species of Aspergillus are responsible for producing aflatoxin. The present study was performed to evaluate the measurement of AFM1, by ELISA method, in milk samples collected from dairy farms of Shahre Ghods, Shahriar (Tehran, Iran).
Material and In this study, during autumn (2016), 82 samples of milk from 41 dairy farms in Shahre Ghods, Shahriar provinces, were randomly selected and assessed AFM1 contamination using the ELISA. On centrifugation of milk, the supernatant including the milk fats was separated and the pellet lacking the milk fat was analyzed through competitive ELISA test, and the amount of aflatoxin was determined.
The results obtained from ELISA assay revealed that 90% of samples were contaminated by AFM1 to a measurable amount and only 8 samples (9.7%) crossed the Iran Standard Level of aflatoxin contamination (50 ng/l).
Milk and dairy products may be contaminated, and since AFM1 is a serious form of threat for human health and is potentially dangerous, it is essential to constantly assess the livestock feed for aflatoxin contamination to minimize or eliminate its amount in milk or dairy products.

Various chemical drugs have been used for leishmaniasis treatment, but their side effects and drug resistance have led to look for new effective compounds. Crataegus microphylla the traditional and medicinal herb is a valuable source of new Pharmaceutical agents. The extract were prepared The extract obtained by maceration method, and diluted with 5% DMSO. Leishmania major promastigotes were cultured RPMI- 1640, enriched with 10% fetal calf serum and Penicillin- Streptomycin.Then the biological activity of herb extract and drug susceptibility was evaluated on L.major promastigotes compared to Glucantime ( Sb III) drug using MTT colorometry. The optical density was measured with Eliza reader set, and the IC50 value was calculated. In this study, we used the GC / mass. IC50 of Glucantime ( Sb III) was 616.18 μg/ml, and alcoholic extracts of Crataegus microphylla 1094 μg/ml. Although Glucantime was more effective than plant extracts, all extracts had profound effects on promastigotes of L.major. The studied herb extract had considerable antileishmanial effects compared to Glucantime ( Sb III) In vitro, the necessity of conducting more experiments to investigate its effect on the parasite in animal model is also appreciated.

A total of 154 samples of marine (n=51) and freshwater fish (n=103) were obtained from fish markets in Elazig Province of eastern Turkey. These samples were tested for Campylobacter, Listeria and Salmonella using culturing and biochemical methods. Campylobacter failed to be detected in any freshwater or marine fish samples. Listeria was detected in 22 and 14 of gill and skin samples from freshwater fish, respectively. L. innocua was isolated at a higher prevalence (14.6%) than L. ivanovii (5.8%) and L. monocytogenes (1%) from the gill samples of freshwater fish. In skin samples, L. innocua was detected at higher prevalence (9.7%) than L. ivanovii (2.9%) and L. welshimeri (1%). However, two (1.9%) of the intestine samples of freshwater fish were found to be positive for L. innocua. In addition, L. monocytogenes isolate yielded a positive band by PCR. Listeria murrayi was the most commonly isolated species with a prevalence of 9.8% and 5.9% from the skin and gill samples of marine fish, respectively. However, the lowest prevalence of L. innocua was found (3.9%) from skin samples of marine fish only, but none of the intestine samples of marine fish were tested positive for Listeria spp. L. monocytogenes was not isolated in any marine fish samples.
• The results indicate that fish can carry a pathogenic Listeria species. However, Campylobacter and Salmonella were not detected in marine fish samples suggests that fish pose no or little risk to the human population in Elazig Province in eastern Turkey.

This contribution reports an ecological benevolent route for the fabrication of copper oxide nanoparticles (CuONPs) using Leucaena leucocephala L. leaves extracts at room temperature. Phytochemical screening of the fresh aqueous leaves extract showed the presence of tannins, saponins, coumarins, flavonoids, cardial glycosides, steroids, phenols, carbohydrates and amino acids. Copper oxide particles such prepared are in Nano scale and their morphology and size are characterized using field emission scanning electron microscopy, energy-dispersive X-ray spectroscopy, transmission electron microscopy, X-ray diffraction, Fourier transform Infra-red spectroscopy, Brunauer-Emmett-Teller, Barrett-Joyner-Halenda and Photoluminescence analysis. Furthermore, CuO-NPs evinced remarkable antimicrobial, antimalarial and antimycobacterial activity against diverse human pathogens.

In this study, the production of aflatoxin B1 (AFB1) and cyclopiazonic acid (CPA) was investigated in toxigenic and non-toxigenic Aspergillus flavus with respect to expression of aflR, veA and laeA genes that are involved to toxins production. A. flavus strains were cultured in YES broth at 28 °C for 4 days and the presence of (AFB1) and (CPA) was confirmed and measured by TLC and HPLC. The expression of aflR, veA and laeA was compared in toxigenic and non-toxigenic strains after cDNA preparation by Real-Time PCR. The results showed that the highest concentrations of AFB1 and CPA were 9450.56 and 403.85µg/g fungal dry weight, respectively. A. flavus isolates based on the ability for producing mycotoxins were divided into 4 groups including, AFB1 and CPA producer (9450.56 and 377.52 µg/g; chemotype I), AFB1 producer (2024.80 µg/g; chemotype II), CPA producer (403.85 µg/g; chemotype III), and non-producer (chemotype IV). The results of the analysis of aflR, veA and laeA gene expression between toxigenic and non-toxigenic A. flavus isolates did not show any significant correlation between the expression of these genes and AFB1 and CPA production among the tested strains in our study. Since, the incidence of AFB1 and CPA producing Aspergillus in environmental pollution is a potential threat to public health, finding the role of related genes of the biosynthetic pathway of aflatoxin and CPA in determining the mycotoxins producing A. flavus is recommended in a larger population for different geographic locations.

Although, many genes are involved in pathogenesis of Campylobacter, racR gene considered the main gene for Campylobacter pathogenicity in humans. The purpose of this study was determined the prevalence of recR gene in Campylobacter spp. isolated from domestic animals and water.
To perform the study 392 fecal and water samples were collected from poultry(182), cow(141), sheep and goat(41) and water(28). All samples were subjected for isolation of Campylobacter spp. using prêt KB method and the presumptive isolates authenticated by DNA sequencing of 16srRNA genes. Finally, Campylobacter isolates assessed for detection of racR gene. The results obtained indicated that 50 strains of Campylobacter spp. were isolated. High isolate frequency (37/50) was for Poultry and low frequency (2/50) was for sheep and goat. Of All isolates thirty six strains were identified as Campylobacter jejuni and the rest (14 isolates) was Campylobacter coli. The genome of 30(83.3%) C.jejuni and 14(100%) C.coli were contained racR gene. Hence based on foregoing evidence racR gene existed approximately in all isolates. Therefore, Campylobacter strains isolated from different sources could be considered infectious agent for human. It is because phenotypical character inducted by racR gene (colonization and heat tolerance) helps them to survive and cause infection.

Shigella is a major cause of dysentery across the world. Appropriate antibiotic treatment of shigellosis depends on resistance patterns. The present study was conducted to identify Shigella species and their antibiotic resistance patterns among dysenteric patients in Rezaei Hospital of Damghan. Isolation of Shigella species was conducted by specific culture medium and biochemical tests. The Shigella species were determined by specific antiserum with agglutination on slide. Then, susceptibility to different antibiotics, i. e. nalidixic acid, ciprofloxacin, ampicillin, tetracycline, co-trimoxazole and ceftriaxone, was tested. The antibiotic susceptibility tests were carried out using the Kirby-Bauer standard method on Mueller-Hinton agar.In this study, 29 Shigella species were found in 91 stool samples of the patients. Determination of Shigella spp. by specific antiserum showed S. flexneri (group B) in 13 cases, S. dysenteriae (group A) in 10 cases, and S. sonnei (group D) in 6 cases, while no case of S. boydii (group C) was found. The antibiotic resistance tests indicated that resistance to co-trimoxazole, tetracycline, ampicillin, nalidixic acid, ciprofloxacin and ceftriaxone was 75.8%, 65.5%, 55.1%, 6.8%, 3.4% and 0% respectively. According to lower resistance to ciprofloxacin and ceftriaxone, it seems that the fluoroquinolone antibiotic, as the first choice, and the third-generation cephalosporin, as the second choice, were suitable for treatment of shigellosis, but regarding the multidrug-resistance likelihood and antibiotic resistance patterns variation in Shigella strains, it is recommended to perform the organism susceptibility test to the antibiotic before treatment.

In recent years, scientists and researchers from all over the world pay much attention to carotenoid pigments as a substitute for antibiotics and synthetic chemical compounds with antioxidant property and the plants, various microorganisms including bacteria and algae are also capable of producing carotenoid pigments. In this research, the production of antimicrobial and antioxidant carotenoid pigment in native strain of Rhodococcus bacterium was studied and evaluated for the first time in Iran. The activation and growth of the native Rhodococcus rhodochrous isolate was done using cultivation on different culture media including Triptic soy agar, BHI agar, ISP5, Bennett's Agar and LB Agar at 30 ° C. Pigment extraction was carried out with different organic solvents and carotenoid pigment components were analyzed using GC-MS method. Antimicrobial activity of the pigment was examined using Agar well diffusion method and its antioxidant property by DPPH(2,2-diphenyl-1-Picrilhydrazil) method was examined and analyzed by comparing to antioxidant BHT. The best media for growth and pigment production were determined TSA and TSB. The antimicrobial activity of carotenoid pigment was obtained against to staphylococcus aureus and Candida aibicans with 21mm and 35mm in diameter of inhibition zone respectively. Antioxidant activity of pigment was determined at the concentration of 8 mg/ml (75.59181 %).GC-MS analyses of extracted pigment showed compounds with the highest peak area. The results of this research indicated that native Rhodococcus rhodochrous strain as a non-pathogenic soil bacterium can be candidate biological source with antimicrobial and antioxidant activity for various applications in farmaucidal and biotechnological products.