Background and aims: Mucorales are fungi belonging to the category of Zygomycetes, found much in nature. Culture-based methods for clinical samples are often negative, difficult and time-consuming and mainly identify isolates to the genus level, and sometimes only as Mucorales. Therefore, applying fast and accurate diagnosis methods such as molecular approaches seems necessary. This study aims at isolating Mucorales for determination of Rhizopus genus between the isolates using molecular methods. Methods: In this descriptive observational study, a total of 500 samples were collected from air and different surfaces and inoculated on Sabouraud Dextrose Agar supplemented with chloramphenicol. Then, the fungi belonging to Mucorales were identified and their pure culture was provided. DNA extraction was done using extraction kit and the chloroform method. After amplification, the samples belonging to Mucorales were identified by observing 830 bp bands. For enzymatic digestion, enzyme BmgB1 was applied for identification of Rhizopus species by formation of 593 and 235 bp segments. Results: One hundred pure colonies belonging to Mucorales were identified using molecular methods and after enzymatic digestion, 21 isolates were determined as Rhizopus species. The sequencing of PCR products and macroscopic and microscopic studies confirmed the existence of R. stolonifera, R. oryzae and R. caespitosus in the samples. Conclusion: Generally, developing a reliable method for determining Zygomycete species can be a useful tool for better understanding of the epidemiology of mucoromycosis.

Toxoplasma gondii, is an isosporean intracellular protozoa in warm blooded animals. Its importance relies on public health hazards and significant economical losses in man and some domestic and wild animals. Its life cycle develops in two hosts: Cat and other felids are definitive hosts which produce oocyts and man and many domestic and wild animals are the accidental intermediate hosts which harbor tissue cysts in their bodies. This parasite causes abortion in man and some herbivorous animals, too. Because the infection rate of toxoplasmosis in sheep aborted fetuses of Ardabil was over 75 %, by PCR method, we decided to use GRA6 gene of Toxoplasma in order to reveal the strains and their pathogenicity in this area. All 75 fetuses were tested by PCR-RFLP method. However, this gene was not amplified in none of the fetuses. Meanwhile, results of bioassay, following intraperitoneal injection of fetal brains to albino mice, revealed that no tissue cyst developed in mice brains. In conclusion, it is probable that the amount of Toxoplasma organisms was not sufficient for the gene amplification. In addition, it seems that the low number of the parasite as well as the presence of non-virulent murine strain of Toxoplasma, may not be the cause of sheep abortion in Ardabil area.

In this study 71 specimens were collected from the Haraz, Shirud and Gazafrud rivers in Mazandaran and Guilan Provinces. The mtDNA was extracted from fish fin using Phenol-Chloroform method. Genetic variation and probable population differentiation of Alburnus chalcoides were studied based on the mitochondrial cytochrome b gene. The specific primers were designed for A. chalcoides and the PCR experiments were performed. Nine restriction endonuclease enzymes were applied for RFLP analysis (BamHI, EcoRV, HaeIII, HincII, HinfI, PpuMI, RsaI, Tsp45I, and TaqI). PCR products (831 bp) and DNA digests were subjected to Agarose gel electrophoresis to separate fragments according to their molecular weight. A part from TagI the rest patterns were identified similar to all specimens from Gazafrud River. Regarding to this patterns, it can be concluded that polymorphism phenomena cannot be observed by above mentioned enzymes and cytochrome b gene, and there is no separate population of A. chalcoides in this study.

In this study 71 specimens were collected from the Haraz, Shirud and Gazafrud rivers in Mazandaran and Guilan Provinces. The mtDNA was extracted from fish fin using Phenol-Chloroform method.Genetic variation and probable population differentiation of Alburnus chalcoides were studied based on the mitochondrial cytochrome b gene. The specific primers were designed for A. chalcoides and the PCR experiments were performed. Nine restriction endonuclease enzymes were applied for RFLP analysis (BamHI, EcoRV, HaeIII, HincII, HinfI, PpuMI, RsaI, Tsp45I, and TaqI). PCR products (831 bp) and DNA digests were subjected to Agarose gel electrophoresis to separate fragments according to their molecular weight. A part from TagI the rest patterns were identified similar to all specimens from Gazafrud River.Regarding to this patterns, it can be concluded that polymorphism phenomena cannot be observed by above mentioned enzymes and cytochrome b gene, and there is no separate population of A. chalcoides in this study.

Background: Trichostrongylus is an intestinal parasite that is highly prevalent in humans and livestock worldwide. There is limited information about the prevalence and epidemiology of Trichostrongylus species among the infected livestock in Mazandaran Province, northern Iran. This study aimed to identify Trichostrongylus spp. among small ruminants using morphometric and molecular methods. Materials and Methods: Small intestinal organs of sheep and goats, slaughtered in Mazandaran Province, were examined for infectivity with Trichostrongylus parasites. Primary species identification was conducted based on the morphological characterization of the male worms. The internal transcribed spacer (ITS) II regions of the ribosomal DNA of the worm tissues were amplified using the polymerase chain reaction (PCR) assay and then the product was subjected to sequencing. Subsequently, the PCR products of the ITS II region were subjected to digestion by HinfI and DraI restriction enzymes using the PCR-restriction fragment length polymorphism (RFLP). Results: Of 180 samples, 98 (54. 44%) were confirmed positive for Trichostrongylus based on the conventional PCR. The digestion of the PCR products with HinfI and DraI facilitated the identification of three Trichostrongylus species, namely Trichostrongylus colubriformis (35%, 90. 81%), Trichostrongylus axei (4%, 4. 08%), and Trichostrongylus vitrinus (5%, 5. 1%). Both morphometric and RFLP techniques resulted in the differentiation of the three Trichostrongylus species. Conclusion: The present study was the 1st attempt in the last 30 years for the identification of Trichostrongylus species in small ruminants in Mazandaran Province. The findings of this study can be helpful for epidemiological and ecological studies, the establishment of effective control programs, and the management of gastrointestinal parasites in Mazandaran Province.

Within the course of this 18-month long work spanning 2010 and 2011, a total of eight human Mycobacterium bovis isolates collected from 1, 320 tuberculosissuspected patients, along with 68 bovine isolates were subjected to PCR-RFLP of pncA gene to assess their resistance to Pyrazynamide. All the isolates were subsequently spoligotyped to better understand their potential epidemiological links.As results showed, only 0.6% of the patients were infected with M. bovis, i.e. a considerably smaller rate compared to previous reports from Iran. Besides, all the isolates proved to be resistant to pyrazynamide. Consulting the SPOLDB4 spoligotyping databank, two patterns, namely ST595 and ST694, were detected among human isolates. Moreover, 12 patterns were found among the bovine isolates. ST595 was the single spoligotype shared between human and bovine hosts. For health officials, these observations indicate that in the Iranian environment M. bovis continues to produce a health risk to human.

Background: In 2009, 3 genome-wide association studies implicated IL28B single-nucleotide polymorphisms (SNPs) as the strongest genetic pretreatment predictor of sustained virological response (SVR) in hepatitis C infection. Recently, the American Association for the Study of Liver Diseases (AASLD) and the European Association for the Study of the Liver (EASL) included IL28B testing in their guidelines.Objectives: The main aim of this study was to develop and validate a simple, rapid, and inexpensive polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for genotyping of common IL28B polymorphisms (rs12979860 and rs8099917).Patients and Methods: Two methods were developed to genotype common IL28B polymorphisms: 1) PCR-sequencing as a reference method and 2) PCR-RFLP as a rapid and inexpensive method. Both polymorphisms were genotyped in 104 Iranian hepatitis C patients by both methods simultaneously. To validate the PCR-RFLP method, the PCR-RFLP genotyping results should be 100% concordant with the PCR-sequencing results.Results: Genotyping of rs12979860 and rs8099917 by PCR-RFLP was concordant with PCR-sequencing in 104 (100%) individuals. The analytical sensitivity and specificity of the PCR-RFLP method for genotyping of both SNPs are 100%. Among these 104 patients with chronic hepatitis C, the frequency of the rs12979860 CC, CT and TT genotypes were 40.4%, 47.1% and 12.5% and the frequency of the rs8099917 TT, GT and GG genotypes were 59.6%, 35.6% and 4.8%, respectively. Also, three IL28B haplotypes (rs12979860-rs8099917) were found among our patients including C-T, T-G and T-T with 63.9%, 22.6% and 13.5% frequency, respectively. C-G haplotype was absent in all of our patients.Conclusions: We have developed a validated, fast, and simple PCR-RFLP method for genotyping of common IL28B SNPs that is more cost-effective than sequencing.

A total of 125 Russian Red Pied cows were genotyped for the prolactim-related gene. The PRL-RsaI genotypes were analysed using the Polymerase chain reaction -restriction fragment length polymorphism (PCRRFLP) method. In this breed, the frequencies of alleles were as follows; A= 0.794 and B= 0.206. The frequencies of AA, AB and BB genotypes were 0.598, 0.392 and 0.01; respectively. Results showed that: BB genotype had higher milk yield than AA and AB individuals (P< 0.05). BB genotype showed higher milk fat yield than AA and AB individuals (P< 0.05). With respect to milk fat content (%), the AB genotype had higher levels than the AA and BB individuals (P< 0.05). No differences between the cows of different PRL-RsaI genotypes were found in terms of milk fat yield and milk protein concentration. The results showed that the highest milk and milk fat yields were obtained by cows with the genotype PRL-RsaI BB. The results presented here demonstrate that the prolactin gene may be considered as a marker for dairy traits in cattle.

Background and Aim: Nowadays, the importance of pathogenicity of non-tuberculosis mycobacteria is well known. Generally, this group, in addition to the respiratory system can cause lymph nodes, skin, soft tissue and bone disorders. Identification of Mycobacterium by culture and biochemical tests may take several weeks and may not be useful for definitive diagnosis. PCR-RFLP (PRA) technique of the hsp65 gene using HaeIII and BstEII enzymes is a precise method for species differentiation, in comparison to phenotypic methods. It is a quick and inexpensive method for detection of mycobacterial species. This study aimed to assess the prevalence of non-tuberculous mycobacteria isolated from the patients referring to tuberculosis center (TB) of Kashan University of Medical Sciences.Material and Method: The study included 106 patients who had been referring to TB Center of Kashan University of Medical Sciences, from 1391 to 1394. The samples were tested by biochemical diagnostic tests. At the same time identification of the strains was made by use of PRA. Amplification of 441-bp fragment was performed by PRA for detection of hsp65 gene.The PCR products were digested with HaeIII and BsteII enzymes and analysis was performed on the basis of electrophoresis.Results: Molecular analysis showed non-tuberculosis mycobacteria in 4 (8.3%) sputum samples, i.e. one positive sample (o.9 %) for every one of the following strains: M. abscessus, M. senegalense, M. fortuitum and M.kansasii.Conclusion: The results of this study showed that some cases of tuberculosis in Kashan are due to non-tuberculosis mycobacteria. Also use of PRA analysis of hsp65 gene for clinical specimens is a rapid and useful tool for identification of species of mycobacterium which is helpful for early diagnosis, treatment and control of tuberculosis.

Although traditional selection for phenotypic values of broiler chickens has made significant improvements in growth rates and meat yields during the past half century, poultry geneticists face additional challenges today, and this is mainly because of negative correlations between production and fitness traits.Accompanying selection for rapid growth, meat-type birds exhibit an increase in physiological disorders. Production performance and fitness traits are negatively correlated in the chicken. Multi-trait selection to improve fitness traits and increase production performance simultaneously is difficult to achieve by using only direct phenotype selection. Molecular marker-assisted selection may be required to increase selection efficiency and make further improvements in production performance. The current study was designed to investigate the associations of AFABP gene polymorphism on chicken growth and body composition traits. So, using PCR-RFLP method, genomic DNAs were extracted from 400 chickens from four different commercial broiler lines. The Taq1 restriction site in exon 1 of A-FABP gene was studied. In addition, the results of PCR-RFLP were verified by DNA sequencing. The comparison of least square means of different genotypes indicated that polymorphism in A-FABP gene was not significant associations (P>0.05) with growth and body composition traits. This research suggests that A-FABP gene could not be a candidate gene that can have an effect on growth and body composition traits in Iranian commercial broiler lines.