Frequency of Antimicrobial-Resistant Genes in Salmonella enteritidis Isolated from Traditional and Industrial Iranian White Cheeses

Iranian white cheese is one of the most important kinds of cheese produced in large scale with high consumption in the country. This dairy product transmits bacterial pathogens like Salmonella spp. Antibiotic resistant Salmonella are widespread in the world. This study was performed to evaluate the frequency of antimicrobial-resistant Salmonella enteritidis and related genes isolated from traditional and industrial Iranian white cheeses. A total of 200 traditional and industrial Iranian white cheeses were collected within Chaharmahal Va Bakhtiari province (southwest Iran). After culturing on specific media using standard bacterial tests the Salmonella sp. was isolated. For specific detection of S. enteritidis from other Salmonella strains sefA gene was studied. Finally, the antibiotic susceptibility patterns were investigated. Results showed that 17 % of cheese samples were contaminated by Salmonella and 5.5 % of specimens by S. enteritidis. The frequencies of resistance genes including tetA, tetB, tetC, cat3, and floR in isolated S. enteritidis were 36.4, 54.5, 81.8, 54.5, and 36.4 %, respectively. All isolated S. enteritidis were susceptible to ciprofloxacin, cefotaxime, and ceftazidime (100 %). In addition, most of them were resistance to chloramphenicol (64 %) and susceptible to gentamicin (98 %). The Salmonella contamination was more frequent in traditional Iranian white cheeses (11.5 %) as compared to industrial (5.5 %) samples (p < 0.05). As compared to industrial samples, high level of resistant genes in Salmonella enteritidis isolated from traditional Iranian white cheeses were observed (p < 0.05). Therefore, traditional Iranian white cheeses are important source of Salmonella contamination in the country hence examination of dairy products for the presence of this pathogen is importan

Investigation of antibiotic resistance and frequency of Clostridium difficile tcdA and tcdB genes in feces of calves in Chaharmahal and Bakhtiari province

زمینه و هدف: کلستریدیوم دیفیسیل یک باسیل گرم مثبت و اسپورزا می باشد که قادر به ایجاد بیماری در انسان و حیوانات است. هدف از مطالعه حاضر بررسی فراوانی ژن های ویرولانس و مقاومت آنتی بیوتیکی کلستریدیوم دیفیسیل جدا شده از مدفوع گوساله ها در استان چهارمحال و بختیاری می باشد. روش بررسی: در این مطالعه مقطعی- توصیفی تعداد 150 نمونه مدفوع تازه گوساله جمع آوری و کلستریدیوم دیفیسیل به کمک روش های کشت باکتریایی جداسازی شد. DNA ژنومی باکتری ها با استفاده از کیت استخراج DNA، تخلیص و ژن های tcdA و tcdB با استفاده از روش Multiplex PCR شناسایی گردید. برای بررسی مقاومت دارویی از روش انتشار دیسک به روش Kirby-Bauer استفاده گردید. یافته ها: تعداد 90 نمونه (60) دارای کلستریدیوم دیفیسیل بوده که از این تعداد، 8 نمونه (8/8 ) دارای ژن tcdA و 16 نمونه (7/17) ژن tcdB داشتند. در آزمایش مقاومت آنتی بیوتیکی مشاهده شد که بیشترین میزان مقاومت، به ترتیب مربوط به آنتی بیوتیک های کلیندامایسن (100) و اریترومایسین (90) است و بیشترین حساسیت به ترتیب مربوط به سیپروفلوکساسین (50) و وانکومایسین (20) می باشد. نتیجه گیری: با توجه به نتایج مطالعه حاضر کلستریدیوم دیفیسیل دارای شیوع و مقاومت آنتی بیوتیکی بالا بوده که می بایست با اتخاذ روش های پیشگیری و درمانی مناسب و همچنین محدود کردن استفاده از داروهای ضد میکروبی در انسان و دام از توسعه ی این باکتری و به خصوص سویه های بیماریزا جلوگیری کرد

The p53 codon 72 polymorphism and association to prostate cancer in Iranian patients

Tp53 is an important tumor suppressor gene, which induces cell growth arrest or apoptosis when subjected to cytotoxic stimuli. Association has been reported between various cancers and p53 codon 72 polymorphism. Our objective was to investigate the possible association between p53 at codon 72 for Arg/Arg, Arg/Pro and Pro/Pro allele polymorphisms in blood samples from 187 prostate cancer patients and 185 controls in southwest Iran by nested-polymerase chain reaction (PCR) of p53 exon 4 and digestion with BstUI restriction enzyme and the DNA fragments were then resolved by electrophoresis in 2% agarose gel. The frequencies of Arg/Arg, Arg/Pro and Pro/Pro genotypes were 39.57% (74/187), 52.41% (98/187) and 8.02% (15/187), respectively, in the cases with prostate cancer, and 27.03% (50/185), 60% (111/185) and 12.97% (24/185), respectively in the healthy controls. Statistically, analyzed and combined results showed there was a significant difference in the frequency of the Arg/Arg genotype and Arg allele between prostate cancer cases and control (p&gt;0.001). These findings suggest that p53 Arg/Arg genotype could be a risk factor for the development of prostate cancer among patients in southwest Iran.Key words: Prostate cancer, suppressor gene (p53) codon 72, polymorphism, Iran

Molecular identification of Ornithobacterium rhinotracheale in turkeys in Isfahan province of Iran

Ornithobacterium rhinotracheale (ORT) is a gram negative, pleomorphic, rod shaped and non-motile bacterial pathogen mostly known to cause respiratory tract infections in turkeys. Ornithobacteriosis is an infectious disease of avian species that has been reported in almost all countries around the world. The objective of this study was to determine the prevalence of ORT in turkeys in Isfahan province of Iran. DNA was extracted from 375 collected tracheal swabs and lungs samples and amplified by ORT 16S rRNA gene specific primers using the PCR technique. ORT DNA was detected in 75 samples (19.93%) of broiler turkeys in Isfahan province of Iran. The results of this study demonstrated the widespread of ORT in broiler turkeys and confirmed that infection with ORT have a high prevalence in Iran.Key words: Ornithobacterium rhinotracheale (ORT), turkey, polymerase chain reaction, Iran

Kappa-casein gene polymorphism in Holstein and Iranian native cattle by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP)

Caseins amount to nearly 80% of the protein output in cow milk. Caseins are biologically important proteins and they are also a raw material for the cheese making industry. The aim of this study was to identify kappa-casein genotype in Holstein and Iranian native cattle. DNA was extracted from 457 blood samples of 247 Holstein and 210 native cattle for identification and genotyping of kappa-casein gene by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) assay using HindIII and TaqI restriction endonucleases. The PCR product of the specific primer K-F and K-R gives the 379 bp specific band. Digestion of 379 bp fragment by restriction endonuclease HindIII generated two fragments of 156 and 223 bp. Result of the cut with this enzyme indicate there genotypes AA, AB and BB in the samples. Also, the amplified DNA (379 bp) from the samples remained undigested by TaqI restriction enzyme. These findings suggest that BB genotype could be a good factor for increase of fat and protein content of milk.Key words: Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP), kappa-casein gene, genotyping, Holstein, native cattle

Occupational risk factors among Iranian farmworkers: a review of the available evidence.

Farming is one of the most important components of most economies. No comprehensive picture exists of the health status of Iranian farmers and the work-related hazards that affect them. We aimed to determine the gaps in the current knowledge regarding the occupational health of Iranian farmworkers. Electronic databases including Medline, Web of Science, Scopus, and Embase, as well as national databases including the Scientific Information Database, MagIran, and Barakat Knowledge System, were searched for articles published through March 2017. All epidemiologic studies regarding the occupational health of farmworkers in Iran were reviewed, regardless of their design, language, time of publication, and location. Of the 86 retrieved articles, 39 studies were ultimately analyzed. Most studies were conducted in Fars, Kerman, and Mazandaran provinces. According to the results of this review, chemical, physical, and biological hazards, along with work-related injuries, may be the main factors threatening the health of farmworkers. The unsafe use of pesticides was related to male infertility, eye and digestive complications, pesticide poisoning, pesticide absorption, hematological changes, non-Hodgkin lymphoma, and multiple myeloma. Chemical hazards (e.g., the unsafe use of pesticides), physical hazards, injuries, and biological hazards (e.g., work-related infectious diseases) threaten the health of Iranian farmworkers. Moreover, farmworkers lack adequate knowledge about the occupational hazards they face and the relevant risk factors

Gene cloning and evaluation of the Acinetobacter baumannii nlpD gene expression in human dermal fibroblast cells using RT-PCR

Background: Acinetobacter baumannii is one of the highly antibiotic-resistant bacteria in the world. This bacterium is a cause of endemic and epidemic nosocomial infections and despite many efforts, there is still no effective vaccine agains it. NlpD is one of the important antigenic agents that stimulate the immune system. So, the aim of this study was to examine gene cloning and expression of the nlpD gene of A. baumannii in human dermal fibroblast (HDF) cells. Materials and Methods: In this experimental study, the nlpD gene was amplified from A. baumannii genome using polymerase chain reaction (PCR). Then, the nlpD gene was cloned and sub-cloned in pTZ57R/T and pIRES2-EGFP vectors, respectively. Confirmation of gene cloning was performed by PCR, restriction endonuclease and sequencing methods. The final pIRES2-EGFP-nlpD recombinant vector was transformed into HDF cells using electroporation and the expression of target gene was evaluated by RT-PCR. Results: In this study, the 831 bp nlpD gene of A. baumannii was amplified successfully. Also, the results of the study showed that the recombinant pIRES2-EGFP-nlpD final construct was produced. Observation of the 831 bp band on agarose gel in transformed cells compared to control cells confirmed the nlpD gene expression in HDF cells. Conclusion: The final construct that generates in this study can express the nlpD gene of A. baumannii in eukaryotic cells. Successful expression of the target gene can be used as a new recombinant vaccine in animal model. The pIRES2-EGFP-nlpD recombinant vector has also the potential as a gene vaccine for future research

Molecular assessment of clarithromycin resistant Helicobacter pylori strains using rapid and accurate PCR-RFLP method in gastric specimens in Iran

Currently, a seven-day, triple-drug regimen has been recommended as one of the first-line therapies for Helicobacter pylori management in which clarithromycin is a key component. Development of clarithromycin resistance leads to the long term assessment of the efficacy of clarithromycin in the triple-drug regimen. The aim of this study was to rapidly and directly assess clarithromycin resistance point mutations on gastric biopsy specimens by using PCR-RFLP method. Biopsy samples were obtained over a 6-months period of 2009, from 200 dyspeptic patients referred to Shahrekord University of Medical Sciences, Iran. Initially, rapid urease test was performed and then DNA was isolated from each tissue and used for molecular analysis such as PCR (for H. pylori diagnostic) and PCR-RFLP (for Cla resistance determination). RUT and PCR results showed that 164 (82%) of the patients were H. pylori-positive. Resistance was evaluated in 164 samples by using enzymes BsaI and MboII. Thirty nine (39) (23/78%) clarithromycin-resistant strains were detected which were identified as 15 (9.15%) A2143G, 15 (9.15%) A2142G and 9 (5.49%) mix strains. The results showed that PCR-RFLP method had a high accuracy to detect A2142G and A2143G mutations associated with resistance to clarithromycin in the minimum possible time.Key words: Helicobacter pylori, clarithromycin resistance, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)