Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343967120120601Development of PCR method for diagnosing of honey bee American Foulbrood diseaseراه اندازی روش واکنش زنجیرهای پلیمراز برای تشخیص بیماری لوک آمریکایی زنبور عسل1510388010.22092/ari.2016.103880ENM. MoharramiH. ModirroustaM. TorkamanJournal Article20160112American foulbrood (AFB) disease is caused by the sporeforming bacterium Paenibacillus larvae larvae. Traditional diagnosis is based on culture technique is time and laboratory work consuming. In this study with standard strain, PCR was developed by specific primers and PCR products were electrophoresed on 0.8 % agarose gel. The PCR primers were selected on the basis of the 16S rRNA gene and amplify a 700-bp amplicon. Detection limits were determined for suspensions of bacteria and spores and also honey and larvae experimentally contaminated. The lowest number of bacteria and spores that were able to detect were respectively 28, 33, 330 and 243 cfu /ml. This PCR technique can be used to identification of the presence of Paenibacillus larvae larvae spores in honey samples, brood samples or on the colonies that grow on medium.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343967120120601A study on Haemophilus influenzae type b growth rate and capsule production in different mediaبررسی میزان رشد و تولید کپسول هموفیلوس آنفلوانزا در محیطهای مختلف71210388110.22092/ari.2016.103881ENM. TorabiA. HaadiE. AsliM. AminianF. EsmailyM. AfsharA. HatamiJournal Article20160112In the present study, seven isolates of Haemophilus influenzae type b were cultured in four different media to compare growth rate and capsule production. Four liquid media namely brain heart infusion broth (BHI), trypticase soy broth (TSB), Mueller Hinton Broth (MHB) and Gonococci broth (GC) with added supplements (1% hemoglobin, 1% Isovitalex) were used. The growth was measured by colony counting (CFU/ml) using serial dilution. Four of the isolates showed the highest growth rate with the average of 1013 CFU/ml on BHI broth while TSB had the second highest growth of more than 1010 CFU/ml in an 18-hour culture at 37 ºC culture. In the next step, the amount of capsular polysaccharide (CPS-b) antigen which is made of Polyribosyl ribitol phosphate (PRP) was assessed all isolates by two methods: modified Indirect Sandwich Enzyme-linked immunosorbent assay (ELISA) and Bial method to select the isolate producing the highest amount of PRP. The antisera used in sandwich ELISA were prepared by immunization of Rabbit and Rat. The maximal amount of PRP was produced by isolate Hib (5s) with the amount of 321 mg/lit.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343967120120601Experimental studies of pathogenecity of Chicken Infectious Anaemia Virus (3 isolates) in Iranمطالعه تجربی پاتوژنیسیته 3 جدایه کم خونی عفونی طیور در ایران131910388210.22092/ari.2016.103882ENM.A. BahmaninejadA. TavasolyR. ToroghiH. MarjanmehrA. ShoushtariA. EzziJournal Article20160112To evaluate pathogenicity of 3 Chicken Anemia Virus isolates-CV1, CV2 and CV3 respectively - the current experiment carried out. After tittering of the viruses, 30 one day old SPF chicks were grouped and intramuscularly inoculated with each isolate in a separate group. Two other groups as positive and control groups were inoculate with a live vaccine virus and normal saline respectively. During the study the birds in first three groups clinically showed raffling feathers, depression and body weight reduction. After 18 days they were weighed, bled and euthanized. Three birds were found dead during the experiment (one in each tested group). Antibodies against CAV in the serum of all 3 groups were detected while no antibody found in negative control group. Hematocrit Values of the three tested groups were under the normal. Grossly the thymus and bursa tissues were severely atrophied. Bone marrow became yellowish and pale. In histopathological study we found aplasia and devoid of erythroid and myeloid cells replacing by fat cells in bone marrow. Severe atrophy and depletion in thymus ,bursa of Fabricius and bone marrow tissues statistically were analyzed and showed that the significance difference in comparing with control groups ( pRazi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343967120120601Immunogenicity of commercial, formaldehyde and binary ethylenimine inactivated Newcastle disease virus vaccines in specific pathogen free chickensبررسی میزان ایمنیزایی واکسنهای نیوکاسل غیر فعال شده به صورت تجاری، فرمالینه و بیناری اتیلن آمین در جوجه های SPF212510388310.22092/ari.2016.103883ENS. Sadigh-EteghadI. KhaliliH. BabaeiR. ToroghiN. RazmaraiiL. FroghyJournal Article20160112Newcastle disease (ND) is one of the most important diseases that affect birds; the epizootic nature of the disease has caused severe economic losses in the poultry industry worldwide. In this experiment ND virus (NDV) was inactivated by two different chemicals binary ethylenimine (BEI) and formaldehyde. Formaldehyde was used at 0.1%, while BEI was used at concentrations of 1 to 4 mM. NDV inactivation with BEI was done in various incubation temperatures and periods and the best result (30 °C, 4 mM BEI and 21 hrs treatment) used as an experimental vaccine. Prepared inactivated NDV vaccines and a commercial vaccine were tested for their efficiency in generating humoral immune response in different groups of specific pathogen free (SPF) chicks. Test groups received 0.2 ml formaldehyde inactivated NDV (NDVF), BEI inactivated NDV (NDVEI) and Razi institute produced NDV vaccine (NDVR) subcutaneously respectively. HI Log 2 total mean titer of NDVEI group (8.42 ± 0.12) were significantly higher than NDVF (7.64 ± 0.16) and NDVR (7.86 ± 0.11) groups (pRazi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343967120120601Stability Study of Razi Trivalent and Monovalent Oral Poliomyelitis Vaccineمطالعه پایداری واکسن خوراکی فلج اطفال سهگانه و تک واحدی موسسه رازی273410388410.22092/ari.2016.103884ENS. Soleimani0000-0002-3914-2909N. BordbarJournal Article20160112Stability studies play a critical role in assuring product quality at all points in the vaccine life cycle. These studies used to determine vaccine expiry date and vaccine efficacy. Accelerated stability and long term stability study performed for three batches of trivalent oral poliomyelitis vaccine (OPV) and three batches of monovalent OPV (type1) manufactured by Razi institute. After sampling, the samples tested intervals in months for long term stability and after 48h incubation in 37 0C for accelerated stability. All of quantitative and qualitative control tests including Potency, Sterility, Mycoplasma detection and Physicochemical testes performed in each period. Potency test for trivalent OPV (tOPV) in three batches of vaccines until 26 months and in two batches, until 27 months after production, met the specification. Potency test for monovalent OPV (mOPV) in all batches of vaccines until 18 month after production met the specification. Sterility, mycoplasma and physicochemical testes on these samples for mOPV and tOPV until expiry date and after that, passed. All batches of vaccines in accelerated stability met the specification after incubation and the reduction titer of vaccines was less than 0.5(-LogCCID50/dose). Results of this research indicated razi polio vaccines are stable for 24 months if the cold chain considered properly.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343967120120601A Case-Control Study of Association between Diarrhea in Newborn Calves and Infection with Rotavirus and Coronavirus in Some Industrial Dairy Herds of Mashhad Aarea, Iran in 2008مطالعه مورد – شاهدی ارتباط بین اسهال گوساله های نوزاد با عفونت روتا ویروسی و کرونا ویروسی در تعدادی از گله های شیری صنعتی اطراف مشهد، ایران354110388510.22092/ari.2016.103885ENE.A. Afshari SafaviG.R. MohammadiM. RadA. NaghibiJournal Article20160112A 1:1 matched case-control study of calves under 1 month of age was carried out by weekly visits to 7 dairy farms in Mashhad from May 2008 to October 2008. Fecal samples were collected from a total of 112 calves with clinical signs of diarrhea and from 112 matched animals without clinical signs of diarrhea as assessed by a scoring system. The samples were investigated for the presence of Rotavirus and Coronavirus by a commercial antigen capture ELISA test. Rotavirus antigen was detected in 29.5% and 17% of diarrheic and non-diarrheic calves, respectively and Coronavirus antigen was detected in 2.7% and 1.8% of diarrheic and non-diarrheic calves, respectively. Among diarrheic calves Rotavirus was the most common in the third week of life and Coronavirus was detected only in some cases of second week of life. The excretion of Rotavirus in the feces of scouring calves was significantly higher than in non-diarrheic calves (P value =0.03, odds ratio = 1.9 (1.05 – 3.76)), but there was not any significant association between shedding of Coronavirus in the feces and diarrhea (P value =0.66, odds ratio = 1.4 (0.24 –9.05)). These results indicate that in these industrial dairy farms in Mashhad, infections by Rotavirus can be considered as an important cause for newborn calf diarrhea.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343967120120601Large scale production of Blackleg vaccine by fermenter and enriched culture medium in Iranتولید انبوه واکسن شاربن علامتی به وسیله فرمانتور و محیط کشت غنی شده در ایران434910388610.22092/ari.2016.103886ENA.R. JabbariR. Pilehchian Langroudi0000.0003.0143.0566M. Moosawi ShoshtariJournal Article20160112In all biological systems growth is defined as increase of chemical compounds. Bacteria can achieve to balanced growth if they are growing in a medium, which are completely adapted to it. Clostridium chauvoei, (Clostridium feseri) is an anaerobic, spore forming, motile, and polymorph bacteria, which its size varies from 0.5-1 to 3-8 micron and could be observed as individual bacterium, diplo, and rarely streptococcus. Blackleg is a fatal disease of young cattle. It produces an acute local infection, and the resulting blood poisoning leads to rapid death. Clostridium chauvoei and, less frequently, Clostridium septicum are the most commonly responsible organisms. Vaccination is the only effective means for controlling of blackleg disease. Several kinds of vaccine are available commercially. It is 4 decades that blackleg vaccine is produced in Razi institute and because of enhanced demand of country, decision was made to improve the production procedure of this vaccine using large-scale fermenter, so the aim of this study was adaptation of Clostridium chauvoei to growth and proliferation in fermenter for preparation of a potent vaccine. Accordingly attempts were made to prepare and formulate the ingredients in order to obtain high yield of Clostridium chauvoei in culture medium by fermenter. All experiments were done in two sets: A-growth in glass bottles using conventional culture medium and B-growth in fermenter using conventional culture medium similar to A and also enriched culture medium. Results showed high yield of Clostridium chauvoei suspension in fermenter after 10 hours, using enriched culture medium (more than 1,480,000,000 organisms/ml), but no significant changes was obtained in glass bottles conditions comparing to the fermenter conditions. The safety and potency of the prepared vaccine was determined in sheep and guinea pigs according to British pharmacopoeia (veterinary) with satisfactory results. Since this research has been successfully done in Razi research institute, so the mono valent inactivated blackleg vaccine, using the enriched culture medium is currently producing by fermenter and is used for immunization of cattles in Iran.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343967120120601Detection of cryoglobulins in serum of Caspian miniature horseمشاهده کرایوگلوبولین در سرم اسبچه خزر515510388710.22092/ari.2016.103887ENJ. JavanbakhtM. KhosraviM. Zamani-AhmadmahmudiN. AtyabiI. DilmaghanianJournal Article20160112Blood samples were collected from 20 healthy miniature Caspian horses at 37 °C. Isolation of cryoglobulin was performed based on a standard method in present study. Harvested sera were kept at 4 °C for two hours and then examined for cryoglubolin. Four serum samples containing precipitate Suspicious of containing cryoglobulin were selected. Subsequently serum protein electrophoresis was performed on normal serum samples and also on four serum samples containing precipitates using an automated electrophoresis system on cellulose acetate strips. In addition Ig isotypes detection (IgG, IgM and IgA) was performed on both precipitates and serum containing precipitates using single radio immunediffusion method (SRID). A narrow-based peak on gamma region of precipitate acetate cellulose strips was observed. Precipitate concentrations were strikingly higher than normal concentration of serum immuneglobulins. It can be suggested that cryoglobulin concentration in a proportion of Caspian miniature horse is higher than other equides may be in relation with animal susceptibility to neoplasias such as lymphoma and leukemia.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343967120120601Virulence assessment of a Neospora caninum isolate for inbred C57BL/6 mouseارزیابی حدت جدایه ای از نئوسپورا کنینوم برای موش اینبرد C57BL/6576110388810.22092/ari.2016.103888ENM.M. NamavariM. SayariM. KargarM. LotfiM. KamalzadehJournal Article20160112Neospora caninum (N. caninum) is an apicomplexan parasite and causes abortion and congenital neosporosis in cattle worldwide. In this study, we evaluate the virulence of a N. caninum isolate on mouse strain C57BL/6. Six groups of five mice C57BL/6 were intraperitoneally inoculated with 1 × l07, 1.5 × l07, 2 × l07, 3 × l07 and 4 × l07 tachyzoites and a control inoculum of DMEM, respectively. Clinical signs and mortality rate were recorded and confirmed by histopathological findings and molecular method. The results of this study indicated that LD50 was 2.5 × l07 tachyzoites of N. caninum per C57BL/6 mouse. This can be used as a lethal challenge model in vaccine development studies.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343967120120601Detection of Colicin genes by PCR in Escherichia coli isolated from cattle in Shiraz-Iranشناسائی ژن کولیسین بوسیله واکنش زنجیرهای پلیمراز در ایکولایهای جدا شده از گاو در شیراز، ایران636710388910.22092/ari.2016.103889ENM. HyatiM. KargarA. PourbakhshZ. ShiraziY. Tahamtan0000-0002-6566-4081M.M. NamvariM. Vesal ShiraziJournal Article20160112A variety of probiotic bacteria have been tested to control animal and foodborne pathogenic bacteria in livestock. The mechanism of inhibition of pathogenic bacteria for several of those probiotic microorganisms is mediated by the production of bacteriocins. Colicins are probably the group of bacteriocins that have been most thoroughly characterized. Colicins are antimicrobial proteins produced by one strains of Escherichia coli to suppress the growth of other strains of E.coli. The present study indicated the preparation of colicin from colicinogenic bacteria. A total of three hundred rectal and rumen swabs isolated from health and diarrheic calves located in Fars province feces. One hundred and fifteen strains were confirmed as E.coli by biochemical test. Polymerase chain reaction was used to determine the following genes encoding colicins. Nearly 100% of isolates were contained at least one gene of colicin. The frequency of several classes of colicin was determined. As a result the most detected gene was Ia Ib and the least detected gene was A.N.S4. Colicin should be tested to control animal and foodborne pathogenic bacteria in livestock.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343967120120601Antibody detection of feline infectious peritonitis virus (FIPV) in sera of companion cats in Ahvaz, south west of Iranجستجوی آنتی بادی ضد ویروس پریتونیت عفونی گربهها (FIPV) در سرم گربههای خانگی اهواز، در جنوب غرب ایران697410389010.22092/ari.2016.103890ENR. AvizehB. MosallanejadM.R. Seyfiabad ShapouriJournal Article20160112Feline infectious peritonitis virus (FIPV) is ubiquitous in domestic cats, especially in young cats and multi-cat environments. In the present study, a total of 248 companion cats of different ages were examined for serum antibody detection of FIPV by immunochromatography assay. The cats were selected from those referring to Veterinary Hospital of Ahvaz University, southwestern Iran from December 2006 to June 2009. Classification was made by age, sex, breed, region and season. The studied cats were divided based on age into three groups ( 3 years) and based on area into five regions (north, east, west, south and central). The results were analyzed by using Chi-square analysis and Fischer's exact test. Seventeen of 248 serum samples (6.85%) had antibody against feline infectious peritonitis virus. Prevalence was significantly higher in young kittens less than 6 months (9.72%; 7 out of 72) and mean-age cats 6 months – 3 years (9.28%; 9 out of 97) compared with above 3 years (1.27%; 1 out of 79) (P0.05). It is necessary to control cat population in these area particular young cats to reduce risk of infection transmission between them.Razi Vaccine & Serum Research InstituteArchives of Razi Institute0365-343967120120601First Report of Coccidiosis and Gizzard Erosion in a Zebra Finch (Taeniopygia guttata) of Iranاولین گزارش کوکسیدیوز و زخم سنگدان در یک فنچ گورخری(Taeniopygia guttata) در ایران757810389110.22092/ari.2016.103891ENH. AzarabadM. NouriM. MoiniJournal Article20160112Coccidiosis and gizzard erosion are rare conditions in cage bird. A male zebra finch was presented with a history of watery diarrhea, anorexia, ruffled feathers, weight loss, and lethargy and died finally. Gross necropsy revealed small areas of erosions and hemorrhages on the gizzard wall. The intestine was oedematous. The spleen appeared pale and small. The testes were asymmetric.Histologically, necrosis of mucosal layer with infiltration of inflammatory cells observed in cecum. Eimeria stages were detected in the enterocytes. In Gizzard, hemorrhage and ulceration of mucosal layer with infiltration of polymorphonuclear cells in to the underlying mucosa were seen. In hepatic tissue, mild focal necrosis with mononuclear cells infiltration was seen. The disease was diagnosed as coccidiosis and gizzard erosion.