Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970120150401Identification of Avian Salmonella Isolates by PCR-RFLP Analysis of a fliC Gene Fragmentشناسایی سالمونلا های جدا شده از ماکیان بر اساس قطعه ژنومی fliC به روش PCR-RFLP1610395610.7508/ari.2015.01.001ENS. Moradi BidhendiF. AlaeiP. Khaki0000-0001-8839-1023R. GhaderiJournal Article20160112The genus of Salmonella is very polymorphic and comprised of a number of genetically closely related serotypes. It is one of the emerging pathogen in food-borne disease which is often found in contaminated chicken eggs. Salmonella enterica is considered one of the major pathogens in public health worldwide. A total of 31 Salmonella isolates identified by specific antisera, which included Salmonella enteritidis (51.6%), Salmonella typhimurium (25.8%), Salmonella infantis (19.4%) and Salmonella colindale (3.2%). DNA was extracted using phenol- chloroform- isoamylalchol method. All the isolates showed fliC gene (1500bp) by using specific primers. PCR products were subjected to digestion using HhaI restriction endonuclease. PCR- RFLP results showed 3 patterns between all isolates. Our research gained in this study demonstrated that using HhaI restriction endonuclease could differentiate Salmonella enteritidis and Salmonella colindale but there is similarity between pattern of Salmonella typhimurium and Salmonella infantis.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970120150401PCR-RELP for detecting of Theileria annulata infection in cattle and Hyalomma species in Kermanshah Province, Iranمطالعه مولکولی PCR-RELP برای شناسایی آلودگی تیلریا آنولاتا در گاو و گونه های هیالوما در استان کرمانشاه71210395710.7508/ari.2015.01.002ENS. SohrabiM. YakhchaliO. GhashghaeiJournal Article20160112Bovine theileriosis is important disease in tropical and subtropical areas with great economic losses in livestock husbandry in Iran. The aim of study was to assess the prevalence of Theileria annulata infection in cattle and Hyalomma species of Kermanshah Province, Iran. A number of 138 blood samples were randomly taken from examined cattle. The genomic DNA was extracted and PCR was performed to specifically amplify a 721-bp-long fragment of the 30 Kilo Dalton major merozoite surface antigen (30 KDa msa) of T. annulata. The amplified products were digested with TaqI, Rasl, and AluI restriction enzymes. Overall prevalence was 9.44% (13/138) with lymphadnopathy (1.17%) and pale mucosal membrane (1.9%) in Holstein cattle agedRazi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970120150401Molecular characterization of Theileria parva parasites from South Sudan using the PCR-RFLP approach on antigen genesشناسایی مولکولی انگل تیلریا پاروا در جنوب سودان با استفاده از آزمون PCR-RFLP بر روی ژن های مولد آنتی ژن132010395810.7508/ari.2015.01.003ENW.L. MarcellinoA.M. El HusseinD.A. SalihD. BerkvensD. GeysenJournal Article20160112 In an attempt to characterize Theileria parva parasites circulating in South Sudan cattle , polymerase chain reaction (PCR)-based assays were carried out using four single copy encoding antigen genes p104, PIM, p150 and p67 in addition to one microsatellite MS321. A total of 20 bovine DNA samples from two locations in South Sudan were included in this study, in addition to two references strains, Muguga and Katete. A total of nine alleles were identified for the polymorphic immunodominant molecule (PIM) locus using restriction fragment length polymorphism (RFLP), against 2 alleles each for the p150 and p104 loci, respectively. This confirms that differences in the polymorphic PIM genes alone could be used to characterize subdivisions in the T. parva populations in the field. On the other hand, 4 alleles were identified for the MS321 locus and 2 alleles for the p67 locus. The data indicate that the studied parasites are genetically closely related, mainly cattle derived and genetically quite distinct from the Muguga strain.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970120150401Isolation and identification of Mycoplasma agalactiae by culture and polymerase chain reaction (PCR) from affected sheep to Contagious agalactia of Khuzestan province, Iranجداسازی و شناسایی مایکوپلاسما آگالاکتیه با استفاده از روش کشت و PCR از گوسفندان مبتلا به آگالاکسی مسری در استان خوزستان212710395910.7508/ari.2015.01.004ENA. AshtariS.A. PourbakhshSh. GhaemmaghamiR. LooniA.R. PooladgarA. Ali ShirudiJournal Article20160112Mycoplasma agalactiae (M. agalactiae) is one of the main causes of contagious agalactia, an infectious syndrome of sheep and goats in Khuzestan province –southwest of Iran that is characterized by mastitis and subsequent failure of milk production, arthritis, abortion and keratoconjunctivitis. This study was carried out to isolation and identification of M. agalactiae with culture and polymerase chain reaction (PCR) method from sheep in Khuzestan province, Iran. A total of 91 samples were collected from milk secretion, eye, ear, nose and joint exudates of sheep. All samples were cultured in PPLO broth supplemented for isolation of M. agalaciae. Extraction of the DNA of bacteria was done by phenol/chloroform method and the PCR assay was applied for detection of Mycoplasma genus in 163bp fragment of 16S rRNA gene and M. agalactiae in 375bp fragment of lipoprotein gene from culture as same as in clinical samples. Out of the 91 samples, 34(37.36%) cultures were shown positive and typical Mycoplasma colonies in PPLO agar culture diagnostic method and 47(51.65%) were scored positive by Mycoplasma genus PCR, 8(8.79%) of the samples were scored positive by using M. agalactiae PCR as diagnostic method. Out of the 91 samples, 26 samples were shown both positive in the culture and PCR, 5 samples were shown both positive in the culture, MPCR and MAPCR. 15 samples were negative in the culture and positive in PCR whereas only 3 samples were positive in culture and negative in PCR. The results showed that the more isolations of M. agalactiae were taken from eye and less in ear and nose samples. M. agalactiae was one of the main factors of contagious agalactia that was detected for the first time from sheep in Khuzestan province.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970120150401The anti-tumor efficacy of nanoparticulate form of ICD-85 versus free formکارآیی و ایمنی فرم نانو ذرات ICD-85 به عنوان یک ترکیب ضد سرطانی در مقایسه با فرم آزاد آن293510396010.7508/ari.2015.01.005ENS. SoheilyA. SarzaemS. MoradhaseliA. Zare MirakabadiH. MorovatiJournal Article20160112Biodegradable polymeric nanoparticles (NPs) have been intensively studied as a possible way to enhance anti-tumor efficacy while reducing side effects. ICD-85, derived from the venom of two separate species of venomous animals, has been shown to exhibit anti-cancer activity. In this report polymer based sodium alginate nanoparticles of ICD-85 was used to enhance its therapeutic effects and reduce its side effects. The inhibitory effect was evaluated by MTT assay. The necrotic effect was assessed using LDH assay. The induction of apoptosis was analyzed by caspase-8 colorimetric assay kit. Cytotoxicity assay in HeLa cells demonstrated enhanced efficacy of ICD-85 loaded NPs compared to the free ICD-85. The IC50 values obtained in HeLa cells after 48 h, for free ICD-85 and ICD-85 loaded NPs were 26±2.9μg ml-1 and 18±2.5μg ml-1, respectively. While it was observed that free ICD-85 exhibits mild cytotoxicity towards normal MRC-5 cells (IC50>60µg ml-1), ICD-85 loaded NPs was found to have higher efficacy in anti-proliferative activity on HeLa cells in vitro without any significant cytotoxic effect on normal MRC-5 cells. The apoptosis-induction mechanism by both form of ICD-85 on HeLa cells was found to be through activation of caspase-8 with approximately 2 fold greater of ICD-85 loaded NPs as compared to free ICD-85. Our work reveals that although ICD-85 in free form is relatively selective to inhibit the growth of cancer cells via apoptosis as compared to normal cells, but nanoparticulate form increases its selectivity towards cancer cells.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970120150401Stability Study of Iriba Brucellosis Full-dose and Reduced-dose Vaccine Produced by Razi Institute in Iranمطالعه پایداری واکسن بروسلوز سویه ایریبا با دز کامل و کاهیده تولید موسسه رازی در ایران374410396110.7508/ari.2015.01.006ENA. EmadiA.M. BehrozikhahS. AlamianS. Soleimani0000-0002-3914-2909E. HasanniaS. DostdariJournal Article20160112 Stability study of biological products plays an important role for determination of product changes in maintenance period and ensuring of safety and efficacy of vaccines. In this research, accelerated and long-term stability study performed for six batches of full and reduced-dose cattle Iriba strain brucellosis vaccine that manufactured by Razi vaccine and serum research institute as a new vaccine. After sampling, the vaccines were tested for accelerated stability, after four days storage at 22 0C and tested intervals in three months until 24 months for long-term stability after storage at 2-8 0C. The result indicated all batches of vaccines in accelerated stability met the specification recommended by OIE 2012 and the mean loss of activity for full-dose was 16.68, 18.87 and 17.79 % and for reduced-dose was 38.85, 36.06 and 34.98 %. In long term stability, the quality control tests including colony forming unit , purity, dissociation and physicochemical tests in all samples until 24 months, met the specification recommended by OIE 2012. The full-dose vaccines showed a mean loss of activity of 30.73, 25.53 and 32.45 % and the reduced-dose vaccines showed 63.51, 58.60 and 60.83 %. The mean increasing of moisture content was, 187.85, 214.13 and 160.77 % for full-dose and 142.35, 110.23 and 164.47 % for reduced-dose. So, the results of this research indicated in spite of moisture content increasing in second year, the brucellosis vaccines with this strain are stable at least 24months if the cold chain considered properly but the best expiry date for the vaccine is one year.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970120150401Detection of Mycoplasma capricolum capricolum from goats of Qom province, Iranشناسایی و ردیابی مایکوپلاسما کاپری کولوم کاپری کولوم از بز های استان قم455010396210.7508/ari.2015.01.007ENS.M. BaraniM.A. BayatzadehA. AshtariA.R. AbtinS.A. PourbakhshE. AsliJournal Article20160112 Mycoplasma capricolum subsp capricolum (Mcc) is one of the etiological agents of contagious agalactia(C.A) in goats which can cause significant economic losses. The aim of this study was to detect Mcc from goats of Qom province in Iran. A total of 111 samples were collected from suspected goats to C.A and cultured in PPLO broth supplemented for Mcc isolation. The bacteria DNAs were extracted from clinical samples and the PCR assay was performed to detect Mycoplasma genus, cluster Mycoides and Mcc from culture. Out of the 111 samples, 33(29.7%) sample cultures were shown positive and typical Mycoplasma colonies in PPLO agar culture method, 53 (47.7%) samples were scored positive by Mycoplasma genus PCR, 8 (7%) of the samples were scored positive by using myciodes cluster PCR and finally 2(1.8%) of the samples were scored positive by using Mcc PCR method. The result of this study showed that Mcc was detected for the first time from Qom province in Iran. Therefore, Qom could be one of the geographical distribution and efficient factors of Mcc in Iranian goats. The lung samples and lymph nodes also could be significant samples for detection of Mcc.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970120150401Detection of Brucella melitensis and Brucella abortus strains using a single-stage PCR methodتشخیص تفریقی بروسلا ملیتنسیس و بروسلا آبورتوس با استفاده از آزمون زنجیره ای پلیمراز تک مرحله ای515510396310.7508/ari.2015.01.008ENN. Nayeri FasaeiA. EtemadiM. EsmaelizadT. Zahraei Salehi0000-0002-5665-5757K. AghaiipoorS. AlamianJournal Article20160112 Brucella melitensis and Brucella abortus are of the most important causes of brucellosis, an infectious disease which is transmitted either directly or indirectly including consuming unpasteurized dairy products. Both strains are considered endemic in Iran. Common diagnostic methods such as bacteriologic cultures are difficult and time consuming regarding the bacteria. The aim of this study was to suggest a single-stage PCR method using a pair of primers to detect both B. melitensis and B. abortus. The primers were named UF1 and UR1 and the results showed that the final size of PCR products were 84 bp and 99 bp for B. melitensis and B. abortus, respectively. Therefore the method could be useful for rapid detection of B. melitensis and B. abortus simultaneously. Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343970120150401A rare case of cutaneous leiomyosarcoma in budgerigar (Melopsittacus undulatus)گزارش یک مورد نادر لیومیوسارکوم جلدی در مرغ عشق576010396410.7508/ari.2015.01.009ENE. KhaksarS.M. NassiriM. Zamani-AhmadmahmudiA.A. SolatiJournal Article20160112 Leiomyosarcoma in birds is relatively rare. This tumor as a muscle neoplasm was reported in captive and free ranging birds. Smooth muscle cells may develop to the leiomyosracoma, but splenic smooth muscle trabeculae is most common site of the tumor growth. Although budgerigars have an incidence of neoplastic diseases but smooth muscle tumors were rarely reported in this species. To the best of our knowledge, present case is the first report of cutaneous leiomyosarcoma in budgerigar. The bird was referred with a history of growing mass in subcutaneous tissue of abdomen. First bird was suspected to the egg impaction, but necropsy confirmed a firm, creamy structure that suspected to the tumor mass. No invasion was observed to the other organs. After excision of the mass, routine histopathologic evaluation and immunohisochemistry investogation were performed for desmin and smooth muscle actin (SMA). Histopathologic examination revealed spindle-shaped cell with cigar-shaped, round, or oval nuclei with cytologic criteria of malignancy including marked pleomorphism and high mitotic activity. These findings were consistent with immunohistochemistry profile of our case and thus confirmed as cutanous liomyosarcoma.