ORIGINAL_ARTICLE Abstract Background and Aims: The Hepatitis C Virus (HCV) is essentially hepatotropic but virus compartment has also been found in other important extra hepatic sites. Detection of HCV RNA in extra hepatic reservoirs such as peripheral blood mononuclear cells (PBMCs) is important for determining of disease progression and effective treatment. The present study aimed to determine of different HCV genotypes in plasma and PBMCs specimens from patients of Yazd province of Iran. Materials and Methods: Blood samples of 50 patients with established HCV referred to Shahid Sadoughi Hospital were collected. These patients had positive anti-HCV and positive plasma HCV RNA. After RNA extraction from plasma and a pellet of approximately 3-5× 106 PBMCs, CDNA synthesis was done and stuck in -70°C. Then HCV genotyping by using of restriction fragment length polymorphisms (RFLPs) method was carried out. Finally, Data analysis was done by SPSS software. Results: The mean age of subjects infected with HCV 38/2±14/6 (ages 21-66) who were the majority of those between ages lower 30 years. 3a and 1a genotype distribution in plasma was significantly higher in those aged over 40 than other age that this difference in genotype distribution at the age of 40 is significant (P value> 0.04) but in PBMCs was not significant (p=0.25). 1a and 3a genotype distribution in plasma and PBMCs in between 40-30 ages showed a lower frequency than other ages. In none of the samples from patients with genotype 2 and 1b genotypes but only 3a and 1a such as mixed infections was detected. Typically, mixing virus infection in 8 patients (16%) in plasma and in 16 samples (32%) was identified in PBMCs. Conclusions: It suggested that plasma subtyping as the target genotype for considering of antiviral therapy is essential, but may be result to the goal of therapy. HCV genotyping in PBMCs samples along with plasma specimen might be beneficial. Therefore, determine of HCV genotype in PBMCs in before therapy because detection of occult infection is useful http://journal.isv.org.ir/article-1-218-en.pdf 2016-09-07 1 7 10.21859/isv.9.2.1 Hepatitis C virus (HCV) HCV genotypes PBMCs Polymerase Chain Reaction Majid Shahidokht a.ahmadi.tums.ac@gmail.com 1 Department of Pathology, Yazd University of Medical Science, Yazd, Iran AUTHOR Mansour Moghimi ahvasmehjani@gmail.com 2 Department of Pathology, Yazd University of Medical Science, Yazd, Iran AUTHOR abbas ahmadi vasmehjani a.vasmehjani@gmail.com 3 Infectious and Tropical Disease Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran AUTHOR Masoud Doosti sajadyaghubi@yahoo.com 4 Infectious and Tropical Disease Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran AUTHOR

ORIGINAL_ARTICLE Abstract Background and Aims: Influenza A(H5N1) viruses  circulating in animals might evolve and acquire the ability to spread from  human to human and thus start a pandemic. Hemagglutinin (HA) has been shown to play a major role in binding of influenza virus to its target cell and the main neutralizing antibody responses elicit against this region. Recent studies have shown that glycosylation of HA is not necessary for its immunogenicity. Bacillus subtilis has been identified as a free endotoxin host for high expression and secretion of heterologous proteins with immunological activity. This bacterium is not capable of supporting glycosylation process. However, it could be an appropriate host to produce new recombinant HA1 for vaccine research purposes. In this study we constructed a recombinant B. subtilis that was able to express and secrete HA1 protein into cytoplasmic and extracellular medium. Materials and Methods: HA1 gene was amplified and cloned into pGEM® 5Zf(–) vector. It was then subcloned into shuttle vector PHT43 and transferred to E.coli for propagation. Accuracy of PHT43-HA1 construct was confirmed by sequencing and restriction map. The recombinant plasmid was extracted from E.coli and used to transform of B.subtilis by electroporation. Following IPTG induction, the total cell protein and the protein secreted into media were analysed through a time course using SDS-PAGE. Results: The accuracy of  PHT43-HA1 construct was confirmed by sequencing and enzymatic digestion analysis. SDS-PAGE results showed that the recombinant HA1 protein was successfully expressed and secreted into medium. Conclusion: The HA1 protein produced here could be considered and evaluated as a protective antigen which its immunogenicity potential needs to be assessed in animal models along with proper control groups. http://journal.isv.org.ir/article-1-180-en.pdf 2016-09-05 8 13 10.21859/isv.9.2.8 Bacillus subtilis hemagglutinin influenza H5N1 Moein Aliakbari m.aliakbari90@yahoo.com 1 Molecular Genetics Department, Research Centre of Biosciences and Biotechnology, Malek Ashtar University of Technology, Tehran, Iran AUTHOR Farida Behzadian fbehzadian@yahoo.com 2 Molecular Genetics Department, Research Centre of Biosciences and Biotechnology, Malek Ashtar University of Technology, Tehran, Iran AUTHOR Ali Asghar Deldar msc_students@yahoo.com 3 Molecular Genetics Department, Research Centre of Biosciences and Biotechnology, Malek Ashtar University of Technology, Tehran, Iran AUTHOR

ORIGINAL_ARTICLE Abstract Background and Aims: Multiple sclerosis (MS) is defined by the presence of inflammatory demyelinating plaques in the central nervous system (CNS). It has been indicated that human herpes virus 6 (HHV-6) may play role in pathogenesis of MS. The aim of this study was to determine the presence of HHV-6 DNA and anti HHV-6 IgG in MS patients and controls. Materials and Methods: Blood samples from 59 patients with MS and 59 healthy controls were collected. DNA extraction and real-time PCR based on (Syber Green) were done in the peripheral blood mononuclear cells (PBMCs) and in plasma specimens. Anti-HHV-6 IgG was measured by ELISA technique. Demographic and clinical data were collected and entered in SPSS 16 and analyzed. Results: HHV-6 DNA was found in the plasma sample of 1 (1.7%) of 59 MS patients. There was no HHV-6 DNA in PBMCs of the MS patients and controls and in plasma of control group. Anti- HHV-6 IgG antibody was present in 84.7% of MS patients and 82.1% of control group. Conclusion: There was no significant relation based on presence of HHV-6 DNA and antibody between MS patients and controls in north of Iran. http://journal.isv.org.ir/article-1-184-en.pdf 2016-09-05 14 19 10.21859/isv.9.2.14 HHV-6 Multiple sclerosis Real-Time PCR ELISA Akramsadat Ahmadi akramsadat.ahmadi@yahoo.com 1 Department of Microbiology, Golestan University of Medical Sciences, Gorgan, Iran AUTHOR Mohammad Hasan Naeimi Dr_naeimii50@yahoo.com 2 Department of Neurology, Sayyad Hospital, Golestan University of Medical Sciences, Gorgan, Iran AUTHOR Naeme Javid naeme_javid@yahoo.com 3 Department of Microbiology, Golestan University of Medical Sciences, Gorgan, Iran AUTHOR Abdolvahab Moradi abmoradi@yahoo.com 4 Department of Microbiology, Golestan University of Medical Sciences, Gorgan, Iran AUTHOR Mohammad Ali Vakili mavakili@yahoo.com 5 Health Management and Social Development Research Center, Golestan University of Medical Sciences, Gorgan, Iran AUTHOR alijan Tabarraei alijant@yahoo.com 6 Infectious Diseases Research Center, Golestan University of Medical Sciences, Gorgan, IR Iran AUTHOR

ORIGINAL_ARTICLE Abstract Background and Aims: Akabane disease causes epidemics of abortions, stillbirths and congenital malformations manifested as arthrogryposis and hydranencephaly or microanencephaly in sheep, goats and cattle. Akabane virus replicates in arthropods and is transmitted by either mosquitoes or midges. Outbreaks of the Akabane virus have been reported from many countries in Southeast Asia, Middle East, Africa and Australia. The aim of this study was to evaluate the prevalence of Akabane virus infection and correlation of this infection with host and environmental determinants in sheep in Khouzestan province, the Southwest of Iran. Materials and Methods: In this study, serum samples of 360 sheep were randomly collected from 6 cities of Khouzestan province and were examined by ELISA assay. Results: Seroprevalence of Akabane virus infection was 39.72% (95% CI: 34.67-44.78%). Statistical analysis showed history of recently abortion and breed of sheep are significantly associated with infection (p<0.05) but sex and age of sheep are not significantly associated with infection (p>0.05). Conclusion: The results of the present study confirm that the Akabane virus infection exists in Khouzestan province, Iran. Considering the local weather conditions and the facility of vector-borne transmission, the health authorities should take measures to prevent and control the infection. http://journal.isv.org.ir/article-1-198-en.pdf 2016-09-05 20 25 10.21859/isv.9.2.20 Akabane virus Epidemiology Prevalence Serology Iran Mohammad Reza Ahi pourmahdim@scu.ac.ir 1 Department of Food Hygiene, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran AUTHOR mahdi Pourmahdi Borujeni reza2dlb@yahoo.com 2 Department of Food Hygiene, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran AUTHOR Mohammad Rahim Haji Hajikolaei mhajih@scu.ac.ir 3 Department of Clinical Science, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran AUTHOR Masoud Reza Seifi Abad Shapouri masoudrs@scu.ac.ir 4 Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran AUTHOR

ORIGINAL_ARTICLE Abstract Background and Aims: EBV is a human herpesvirus that infects more than 90% of the world’s population. Although, benign and asymptomatic in most cases, the infection can cause many nonmalignant and malignant disorders of lymphoid and epithelial origins. The objective of this study was detected in IM prevalence in Neyshabur, Northeast Iran from 2010-2014. Materials and Methods: This cross-sectional descriptive epidemiological survey was performed in Neyshabur between 2010 and 2014 to reveal the prevalence of infectious mononucleosis. A total of 114 patients were studied. Briefly, patients with a positive test for specific IgG and IgM were determined as positive cases. Results: the overall prevalence of IM was 14%. The mean age±SD for infectious mono test is 18.96± 15.79. the age groups of 0-10 and 21-30 years old, were the most positive cases in this period. In addition, 31-40 and upper 50 years were not positive cases. Male patients were significantly more positive and likewise, it was observed that the spring and summer seasons had significantly higher positive cases of IM. Among the five years of this study, it was a decreasing status from 2010 (23.1%) to 2014 (9.1%), although a slight fluctuation was occurring. Conclusions: the prevalence of IM was low in Neyshabur city. Moreover, children and male patients had relatively higher prevalence of the disease. Furthermore, it was observed a higher rate of IM in the spring and summer seasons. http://journal.isv.org.ir/article-1-175-en.pdf 2016-09-09 26 30 10.21859/isv.9.2.26 Epstein-Barr virus infectious mononucleosis Neyshabur city Iran M Salehi 1 Medical Diagnostic Laboratory of Neyshabour, Center of Medical, Pathological and Genetic Diagnostic Services, Iranian Academic Center for Education, Culture and Research (ACECR), Mashhad Branch, Mashhad, Iran AUTHOR S Kh Shokuhi-Mostafavi 2 Department of Microbiology, Tehran Medical Sciences Branch, Islamic Azad University, Tehran, Iran AUTHOR M Mirzaee 3 Department of Laboratory Sciences, Borujerd Branch, Islamic Azad University, Borujerd, Iran AUTHOR M Mobini 4 Young Researchers and Elite Clup, Islamic Azad University, Tonekabon Branch, Tonekabon, Iran AUTHOR M Gholami 5 Isfehan University, Isfehan, Iran AUTHOR Asghar Heydari M 6 Isfehan University, Isfehan, Iran AUTHOR

SHORT_COMMUNICATION Abstract Background and Aims: Infectious bronchitis virus (IBV) is the causative agent of avian infectious bronchitis, which is characterized by respiratory, reproductive, and renal signs. IBV is a highly variable virus with a large number of genotypes. S1 gene sequencing has been used for molecular epidemiological studies and genotypic characterization of IBV. To better understand the molecular epidemiology of IBV in Iran, we sequenced the S1 gene of IBV field isolates, a total of 40 tracheal and kidney tissue specimens from different commercial broiler flocks in the East of Iran were collected from 2015.  Materials and Methods: 15 IBV-positive samples were selected from among the total and were further characterized by sequencing the spike glycoprotein gene.   The isolates were confirmed by real-time polymerase chain reaction (PCR) and characterized by sequencing the spike glycoprotein gene. Results: Three genotypes were detected. The percentage of Variant 2 (IS/1494 like), 793/B, and QX genotypes was 66.7%, 26.7%, and 6.6% respectively. We reported the QX as the first time and Variant2 was the dominant genotype in this area. Conclusion: It is an updated and comprehensive study of genotyping of IBV and completes IBV puzzle in the East of Iran. http://journal.isv.org.ir/article-1-264-en.pdf 2017-02-08 31 35 10.21859/isv.9.2.31 Avian Infectious Bronchitis Iran Genotyping Spike A Ghalyanchi-Langeroudi 1 Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran AUTHOR V Karimi 2 Department of Avian Medicine, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran AUTHOR A Jannat 3 Department of Avian Medicine, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran AUTHOR M Hashemzadeh 4 Department of Research and Production of Poultry Viral Vaccine, Razi Vaccine and Serum Research Institute, Karaj, Iran AUTHOR MH Fallah 5 Department of Research and Production of Poultry Viral Vaccine, Razi Vaccine and Serum Research Institute, Karaj, Iran AUTHOR F Gholami 6 Department of Avian Medicine, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran AUTHOR MT Zabihi 7 Department of Avian Medicine, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran AUTHOR M Heidarzadeh 8 Iranian Veterinary Organization, Zahedan , Iran AUTHOR