Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970320151001Molecular characterization of the lipL41 gene of Leptospira interrogans vaccinal serovars in Iranآنالیز مولکولی ژن lipL41 در سرووارهای واکسینال لپتوسپیرا اینتروگانس در ایران14515010397310.7508/ari.2015.03.001ENM.S. SoltaniP. Khaki0000-0001-8839-1023S. Moradi BidhendiM.H. ShahhosseinyJournal Article20160112Leptospirosis caused by infection with pathogenic leptospires, which is the most prevalent zoonotic disease in the world. The outer membrane proteins (OMPs) of pathogenic leptospires such as LipL41 play a crucial role in pathogenesis of this disease. Therefore a major challenge to develop an effective vaccine against leptospirosis is application of basic research on the OMPs of leptospires to improve vaccine development. The aim of this study was cloning and analyzing of the lipL41 gene from vaccinal serovars of leptospires in Iran, in order to identify genetic conservation of this gene. Three vaccinal serovars of Leptospira were used in this study. The lipL41 gene of these serovars were amplified and cloned in the pTZ57R/T vector. The recombinant clones were confirmed by colony-PCR and sequencing. The sequenced genes were analyzed for their homology between them and other submitted sequences in Genbank database using the BLAST and MegAlign program. PCR amplification of the lipL41 gene resulted in the 1065 bp gene product in vaccinal serovars tested. In our study, nucleotide sequencing results showed high similarity (>94%) within the leptospiral vaccinal serovars. The genetic conservation of the lipL41gene among different serovars of Leptospira indicated the capacity of utilization of this gene for development of recombinant vaccine against leptospirosis.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970320151001Genotyping of Pasteurella multocida ovine and bovine isolates from Iran based on PCR-RFLP of ompH geneتیپ بندی جدایه های پاستورلا مولتوسیدای ایرانی بر اساس ژن کد کننده OmpH15115610397410.7508/ari.2015.03.002ENA. GhanizadehPasteurella Research Laboratory, Razi Vaccine and Serum Research Institute, Karaj, IranA.R. JabbariPasteurella Research Laboratory, Razi Vaccine and Serum Research Institute, Karaj, IranJ. ShayeghDepartment of Veterinary Microbiology, Faculty of Veterinary, Shabestar branch, Islamic Azad
University, Shabestar, IranA. SanchuliPasteurella Research Laboratory, Razi Vaccine and Serum Research Institute, Karaj, IranR. BanihashemiPasteurella Research Laboratory, Razi Vaccine and Serum Research Institute, Karaj, IranJournal Article20160112Pasteurella multocida (P. multocida), A Gram-negative facultative anaerobic bacterium, is a causative animal pathogen in porcine atrophic rhinitis and avian fowl cholera. The outer membrane of Gram-negative bacteria contains of many different protein in very high copy numbers. One of the major outer membrane, the H proteins have functional as high immunogenicity and antigenicity. In this study to increase information about epidemiology of ovine and bovine P. multocida, the 24 isolates from sheep and nine isolates from cattle were investigated by PCR-RFLP analysis of the ompH gene. In all 33 isolates, digestion of the amplified fragment of ompH gene by using EcoRI, cfoI and HindIII produced 3, 5 and 3 different restriction patterns respectively. Sixteen RFLP patterns were found among 33 investigated P.multocida isolates. This study showed that, the PCR RFLP based on ompH gene is potentially a useful method for typing of P. multocida isolates from sheep and cattle. The RFLP patterns of this gene exhibited extensive restriction site heterogeneity, which may be particularly suitable for fingerprinting of P. multocida isolates. Considering ompH protein as a protective immunogenic moiety of P. multocida, the results of this study showed a heterogenic bacteria and this means the possibility to produce a multivalent vaccine to be protective against diseases caused by this organism in sheep and cattle in Iran.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970320151001Peptide based polyclonal antibody production against bovine rotavirus non structure protein4 (NSP4)تولید آنتی بادی پلی کلونال بر اساس پپتید علیه پروتئین غیر ساختاری NSP4) 4) روتا ویروس گاوی15716110397510.7508/ari.2015.03.003ENH. RazaviA. TeimooriE. SaberfarH. Soleimanjahi0000-0003-1931-7801Z. GoodarziJournal Article20160112The rotavirus nonstructural protein 4 (NSP4), is a multi functional protein that play key role in both viral morphogenesis and cytopathic effect associated with cell death. However, the complete biological effect of NSP4 remains to be clarified. Since to obtain further knowledge about this protein there is a need for <br />recognizing antibody and there is no commercial antibody against this protein, this study was designed to produce polyclonal antibodies against a synthetic immunogenic peptide of RF rotavirus NSP4 protein in order to be used as diagnostic and research tool. In the present study a peptide sequences corresponding to <br />NSP4 114-135 conjugated to Bovine Serum Albumin (BSA) by glutaraldehyde through a single-step coupling protocol. The conjugated peptides were extensively dialyzed and injected to New Zealand white rabbits. The NSP4 114-135 peptide-specific antiserum was confirmed by both IF and Western blot analysis. <br />The result indicated successful production of polyclonal antibody raised against native NSP4 protein. In conclusion, since NSP4 is a toxic protein for the cells, it is impracticable to express full length of NSP4 protein in expression systems. Therefore, the introducing immunogenic peptide in appropriate animal may <br />be the best approach to produce its specific antibody.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970320151001Development and Cytogenetic Characterization of a Continuous Bovine Kidney Cell Line (IRKHBK) and Evaluation its Susceptibility to some Virusesتولید و تعیین مشخصات ژنتیکی تیره یاخته ای مداوم کلیه گوساله(IRKHBK) و ارزیابی حساسیت آن نسبت به برخی از ویروس ها16316910397610.7508/ari.2015.03.004ENS. MasoudiJournal Article20160112In this syudy a continuous bovine kidney cell line derived from a primary bovine kidney cells was established for the first time in Iran. The cells were originating from two-day-old normal male calf of Holstein breed. The cell cultures were continuously passaged following complete proliferation of primary <br />cells. The specific properties or characteristics of the cell were defined using cytogenetic and tumorigenicity analysis. An increasing in cell proliferation was observed at 30th passage. Subsequently chromosomes analysis was shown the first chromosomal adhesion. In karyotyping a decrease in number of the cell¢s <br />chromosomes (n=59) was detected compared to the normal bovine cells chromosome count (2n=30). The cell obtained unlimited proliferation capacity from 70th passage and was identified as infinite cells at passage 90th. The continuous cell line named Iran Razi Khedmati Bovine Kidney (IRKHBK) and was <br />deposited in National Cell Bank of Iran (NCBI), Pasteur Institute. Susceptibility of the IRKHBK cell line for isolation and replication processes of bovine herpesvirus-1 (BHV-1) and bovine virus diarrhea-mucosal disease (BVD-MD) were also evaluated. The results showed that this cell is more susceptible to the viruses compared to primary bovine kidney cells. According to our results, IRKHBK cell is recommended for routine assays of viruses as a substitution for primary bovine kidney cells.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970320151001Study on phenotypic characteristics of Salmonella gallinarum and Sallmonella pullorum isolates based on biochemical and antimicrobial susceptibility tests in Iranمطالعه خصوصیات فنوتیپی جدایه های سالمونلا گالیناروم و سالمونلا پلوروم بر اساس آزمایشات بیوشیمیایی و تعیین حساسیت ضد میکروبی در ایران17117710397710.7508/ari.2015.03.005ENP. S.KhakiS. Moradi BidhendiN. CheraghchiJournal Article20160112Salmonellosis is a very important disease of avian species because of its huge economic impact, worldwide distribution and difficulty posed in its control. Fowl typhoid and pullorum disease, is caused by Salmonella enterica subsp enterica serovar Gallinarum biovar Gallinarum and Pullorum. The purpose of this study was <br />to investigate the biochemical characteristics and antimicrobial susceptibility of Salmonella gallinarum and Salmonella pullorum. A total of 13 Salmonella isolates, identified by biochemical tests and specific antisera including Salmonella gallinarum (n=10) and Salmonella pullorum (n=3). All were found to be susceptible to gentamicin. Also 7 (53.8 %), 6 (46.1%) and 5 (38.4%) isolates were resistant to streptomycin, cephalexin and nalidixic acid respectively. Multidrug resistance to three or more antibiotics was observed in 6 (46.1%) isolates and overall 9 antibiotic resistance patterns were recorded. The results showed that poultries as a source of antimicrobial resistance could pose a serious risk to public health via food chain transfer. Hence more epidemiological surveillance programs and antibiotic susceptibility investigations are advised.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970320151001Detection of Shiga toxin-producing Escherichia coli (STEC) in faeces of healthy calves in Mashhad, Iranشناسایی اشریشیا کلی تولید کننده شیگاتوکسین (STEC) در مدفوع گوساله های به ظاهر سالم در شهرستان مشهد، ایران17918510397810.7508/ari.2015.03.006ENA. JamshidiM. RadT. Zeinali0000-0003-1596-1544Journal Article20160112The aim of this study was to identify virulent Shiga toxin-producing Escherichia coli (STEC) strains isolated from faecal samples of 100 clinically healthy calves. In the present study, a total of 100 Escherichia coli (E. coli) isolates from clinically healthy calves belonging to 6 different farms located in Khorasan Razavi province, Iran, were examined for presence of virulence genes characteristic for STEC strains. Duplex PCR assay was used in order to determine the presence of the stx1, and stx2 genes. The stx1 or stx2 positive isolates was later analyzed with primers specific for eaeA, hlyA genes. PCR assay was also conducted on these isolates for detection of O157: H7 serotype. Our results showed that 15 isolates (15%) were positive for stx1 gene, 19 isolates (19%) were positive for stx2 gene and 8 isolates (8%) were positive for both stx1, and stx2. Totally 26 isolates were positive for at least one of stx1or stx2 genes. Among 26 isolates, which were tested for the presence of eaeA and hlyA genes, 5 and 22 of them were positive for these genes, respectively. Four isolates were positive for both eaeA and hlyA genes. Among 26 isolates which were analyzed for genes coding O157 and H7 antigens, one and four isolates were positive for O157 and H7 antigen gene, respectively. None of the isolates were positive for both antigen genes. Faecal contamination due to poor hygiene is a risk factor in contamination of meat and milk products.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970320151001Effects of climatic factors on prevalence of developmental stages of Fasciola gigantica infection in Lymnaea snails (Lymnaea auricularia var rufescens) in Bangladeshمقاله خارجی فاقد چکیده فارسی می باشد نویسندگان خارجی هستند18719410397910.7508/ari.2015.03.007ENK.M. IslamM.D. IslamS.M.A. RaufA. KhanM.K. HossainS. SarkarM. RahmanJournal Article20160112The present study was conducted to investigate the prevalence of developmental stages of Fasciola gigantica infection in Lymnaea snails and their populations in Sylhet region of Bangladesh. A total of 1865 Lymnaea snails were collected and examined, of which 56 (3%) were found to having infected with developmental stages of Fasciola gigantica. Only 4.08% infected of 515 snails were collected from Biswanath Upazilla followed by 3.16% of 443 from Beanibazar, 2.53% of 396 from Balaganj then 2.40% of 292 Jaintapur Upazilla and the lowest 1.83% of 219 from Sylhet Sadar. Month-wise data, the prevalence of snail infections was observed to be the highest in May (5.06%) and August (5.61%) and the lowest in March (0.74%) and February (0.68%). However there was no infection observed through November to January. Seasonal prevalence of the developmental stages of F. gigantica infection in Lymnaea snails was also studied. Highest prevalence (4.63%) was recorded during rainy season and lowest prevalence (0.76%) was recorded during winter season. The study revealed that the developmental stages of F. gigantica infection in snail populations decreases from November to January and increases from February to October and highest in August and September in Sylhet region of Bangladesh.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970320151001Molecular detection of the infection with Fasciola hepatica in field-collected snails of Galba truncatula and Lymnaea stagnalis from West Azarbaijan, Iranشناسایی مولکولی آلودگی حلزونهای گالباترونکاتولا و لیمنه استاگنالیس به فاسیولا هپاتیکا در استان آذربایجان غربی19520210398010.7508/ari.2015.03.008ENR. Malekzadeh-ViayehA. Imani BaranM. YakhchaliJournal Article20160112The liver fluke, Fasciola hepatica, is considered as the most common cause of fasciolosis in both domestic livestock and human. This study was carried out to detect the prevalence of the larval stages of F. hepatica in the snails Galba truncatula and Lymnaea stagnalis in West Azarbaijan, Iran. Snail collection was performed through searching 28 freshwater habitats from May to December 2010. Following the identification of the two snail species, polymerase chain reaction (PCR) was utilized to amplify the 28SrRNA gene of F. hepatica in the snails’ tissues. The amplified DNA fragment was subjected to restriction fragment length polymorphism (RFLP) analysis. According to the RFLP patterns, 16.6% of the examined G. truncatula and 1.1% of L. stagnalis were infected by F. hepatica. While there was not detected infection with larval stages of F. gigantica in any examined snails. The RFLP analysis of 28SrRNA gene was proven to be a useful tool for detection of the infection and its transmission by the intermediate hosts, and can help with the establishment of suitable control programs against fasciolosis in livestock and human in any region of interest.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970320151001Isolation of Ornithobacterium rhinotracheale from the brains of commercial broiler breeder chickens with meningitis and encephalitisجدا سازی اورنیتوباکتریوم رینو تراکئال از مغز مرغان مادر گوشتی با علائم مننژیت و آنسفالیت20320910398110.7508/ari.2015.03.009ENM. BananiM.H. HablolvaridR. MomayezA. NouriN. GhodsianA. AshtariS.G. MirzaeiJournal Article20160112Ornithobacterium rhinotracheale (ORT) has been identified as one of the respiratory bacterial pathogens in turkey and chicken flocks. Four live birds displaying severe torticollis were submitted from a 13-week-old commercial broiler breeder chicken flock located in Mazandaran province. These birds were suspected to <br />pasteurellosis by the farm veterinarian. No other marked gross lesion except emaciation was seen. Histopathologic examination of the brains showed mild to moderate meningeal vasculitis, perivascular cuffing with lymphocytes, degeneration and necrosis of purkinje cells in the cerebellum. Viral culture of the brains <br />especially for Newcastle disease and avian influenza viruses was negative. Bacterial culture of the brains onto the blood agar revealed pure growth of Ornithobacterium rhinotracheale. In this study molecular confirmation of ORT by using of a very specific polymerase chain reaction (PCR) was carried out. Amplification products of a 784 bp region of the 16S rRNA gene of ORT confirmed the bacterium identification. This is the first field case of ORT isolation from the brain of commercial chickens in Iran. These data suggest that this bacterium should be considered in differential diagnosis in cases of avian nervous signs. Further studies are necessary to confirm if ORT is a primary pathogen in such cases.