Evaluation of the efficacy of a new oil-based adjuvant ISA 61 VG FMD vaccine as a potential vaccine for cattle

Foot-and-mouth disease is an important viral disease of cloven-hoofed animals. Inactivated whole particle virus vaccines are still widely used in prophylactic vaccination campaigns. The choice of adjuvant is a very important factor in enhancing immune responses and the efficacy of inactivated vaccines. Montanide ISA 61 VG is a new ready-to-use mineral oil-based adjuvant developed by SEPPIC Inc. [SEPPIC, France] with high-potential immune responses needed for clinical protection against FMD infection. In this study, we compared the efficacy of two FMD vaccines either formulated with the new oil-based adjuvant ISA 61 VG and saponin, or with aluminum hydroxide gel and saponin. Both vaccines contained the same antigen payloads of O2010/IR. Two groups of 15 naive cattle received a single vaccination with different doses [full dose, 1/3 dose and 1/9 dose] to calculate their PD50 [50% protective dose] after being challenged with the homologous virulent virus. The mean neutralizing antibody titer was determined at 0, 7, 14 and 21 days after vaccination, measured by a micro neutralization test. The new vaccine improved humoral immune responses by 19%, while inducing a higher geometric mean. The titer for neutralizing antibodies was 2.91 log10 compared to the alum-gel based adjuvant vaccine which was 2.44 log10 [P-value=0.1782]. The new vaccine showed a PD50 value of 10.05 as compared to a PD50 value of 4.171, respectively. According to the results, the FMD vaccine formulated with the new oil adjuvant, ISA 61 VG, shows potential as an alternative vaccine for routine and emergency vaccinations in the FMD enzootic region

[Molecular typing of group Abovine rotavirus of calves in the[provinces of Tehran, Alborz and Qazvin]]

BACKGROUND: Group Abovine rotavirus [BRV-A] is one of the most important causes of gastroenteritis and diarrhea in newborn calves

OBJECTIVES: Major types of BRV-A in Tehran, Alborz and Qazvin were detected in this study

METHODS:Atotal 125 fecal samples of calves showing clinical signs of diarrhea were collected from 26 industrial dairy farms located in the provinces of Tehran, Alborz and Qazvin, during two years

RESULTS: BRV-A was detected in 39.2 % [49/125] of total samples using a commercial ELISA kit. Thirty five positive samples were analyzed by seminested multiplex RT-PCR for Pand G genotyping. G10 was the most prevalent genotype, accounting for 57.1% of samples, G6 accounted for 22.9% of samples and in 5.7% of samples [2/35], mixed infection of both genotypes G6 and G10 were detected. Also, the detected P types were P[11] and P[5], accounting for 71.4% and 14.2%, respectively. In our study, none of the genotypes G8 and P[1] were detected. The incidence of genotype combinations corresponded to the B223-like strains [G10P[11]], UK-like strains [G6P[5]] and KN4-like strains [G6P[11]] were 51.4%,14.3% and 8.6%, respectively. Mixed infections G6/G10P[11] were detected in 2.85% of all samples analyzed with RT-PCR

CONCLUSIONS: To the best of our knowledge, this is the first report about the determination of Pgenotypes of BRV-Aand distribution of the most common BRV-Astrains circulating in Iran

Our study also indicated that the incidence of the G genotypes of BRV-Ain the provinces of Tehran, Alborz and Qazvin, which is one of the greatest husbandry centers in Iran, has changed in the past years. Furthermore, this finding could be valuable in rotavirus vaccine design

[Molecular epidemiology of foot-and-mouth disease virus in ruminant during 2005-2006 in Iran]

Foot and mouth disease [FMD] is a highly contagious viral disease of ruminant which causes fever and postule on mouth, hoof and teat. The main purpose of this study was to determine and charachtrize isolated FMD virus from Iran between 2005-2006, and to compare it with vaccine virus strains. After preparation of samples, serological test for typing of virus was performed. In order to virus isolation. the samples were inoculated to IBRS2 cell, RT-PCR and PCR were used for sequencing. Two dimentional virus neutralization test was carried out for detecting of immunological relationship [r value] between the field isolate and virus presented in vaccine. Detected strains were as follows: 241 samples of type AO5IR, 125 of type A87IR, 79 of type 0.3 of type Asia and 714 negative out of 1162 samples. Average r-values of type A, 0, Asia field virus with vaccine strains were 50-92% and 97%, respectively. Phylogenic tree was designed according to the nucleic acid sequencing data. There is not strong relationship between field viruses of type A and vaccine viruses. However a strong relationship was shown for type 0 and Asia ones with vaccine virus strains

Study of effector antigens of IBR virus on humoralimmunity

The objective of this study was to determine the protein pattern and antigenic structure of BHV-1. Ten viral isolates from cows with clinical signs of respiratory infections were selected for this study and confirmed by SN test. All samples were cultured on cell cultures and purified by ultracentrifugation. Purified isolates were electerophorsed using SDS-PAGE method and immunoblotted. Viral antigens were detected using anti BHV-1 bovine antiserum and molecular weight of the bands were determined. All samples showed identical band number and molecular weight except for one sample which revealed an additional 120-125 KDa band. Also immunoblotting data resulted in identified of twelve bands [150, 130, 115, 97, 77, 70, 55, 45, 40, 38, 32 and 25 KDa] suggesting involvement of these antigens in evoking humoral immune response in affected cattle

[ survery on frequency of foot-and-mouth disease virus carriers of slaughtered cattle in Zyaran abattoir]

To determine frequency of FMDV carriers in slaughtered cattle at Zyaran abattoir. Cross-sectional study. Three hundred and one tonsils of clinically normal cattle. All of tonsils were tested for determination of FMDV genome using RT-PCR. Then, 30 samples were cultured on MDBK. 20 samples had positive findings in RT-PCR test. Based on sex, age and breed of cattle, RT-PCR results compared with Chi-square and Fisher's Exact Test. McNemar test was used for comparing of virus culture results with RT-PCR. Ninety nine samples were positive in RT-PCR. Relative frequency of FMDV genome presence in tonsils of clinically normal cattle had a significant difference based on sex and breed. The Sistani and Holstein bred had the highest and the lowest relative frequency, respectively samples on cell serotype O of culture were FmDV. One positive sample [Asia 1] Two positive sample[Asia1] was shown by ELISA. Total relative frequency of positive FMDV genome [32.9%] indicates the extensive exposure to FMDV. Breed variation among Sistani cattle [as a variety of Bos indicus] and other bred should be studied in a controlled condition

Molecular diagnosis of Marek's disease virus [MDV] in Iran

The purpose of this study was diagnosis of Marek's disease virus as one of the causative agents for visceral tumors in chickens using Polymerase Chain Reaction. Forty blood samples from the chickens without any clinical signs of Marek's disease and another 42 tumoral tissues from commercial chickens were collected. The whole DNA of the samples were extracted using a silica gel DNA extraction kit, then PCR test was performed using specific primers detecting 132bp tandem repeat and antigen A gene of MDV, finally electrophoresis of PCR products was done in Iko[3]/41 agarose gel. No positive results were obtained in blood samples for MDV and it's vaccinal strain, but about 47.6% of samples were positive. The tumoral tissues including liver, spleen, proventriculus, ovary, breast muscle and bursa of Fabricius. The vaccinal strain of MDV [Rispens] was not detected in any of examined blood samples, as the period of viremia for this virus is very short. Serotype 1 of Marek's disease virus was detected as a causative agent of tumors in the chicken farms of Iran as a first step in this study

[ study on cellular changes in inoculated cell cultures with Pestivirus, using direct immunoflurescence technique]

Objective: this study was carried out to find the progressive cellular changes developed in cell culture with cytopathic [cp] and noncytopathic [ncp] pestiviruses isolated from cow and sheep

Design: observational study

Procedure: following inoculating the R-BK cell cultures in Lighton tube with NADL strain of BVD, a cp strain of Border disease virus and a nocytopathic BVD, the cells were stained with FA and observed with IF microscope. In inoculated cultures with cp and ncp pestivirus obvious changes were present at the first day and multiple focal expression of viral antigens were seen

Results: in the second day in cp inoculated cultures, the morphological changes began and antigenic spread were more intensive. In the 3rd day the CPE foci were seen in different parts of the cultured cells with high concentration of antigen around the CPE sites. The cells started to detach and became rounds and finally in the 4th and 5th day, almost all cells were antigen positive and developed CPE. However, those cultures inoculated with noncytopattuc strain of BVD, in daily observation the antigenic spread was too slow and up to 6 days post inoculation no CPE were observed

[Detection of infected goats and sheep, with Cpseudo-tuberculosis via seroagglutination]

In this survey a total of 953 serum samples [sheep and goats] were collected from Ziaran and Varamin slqughter houses. The seroagglutination test was carried out with C.ovis antigen which was prepared in Microbiology Dept. A positive control serum was made on rabbit after several injection with C.ovis. The goats and sheeps with the titer higher than 1/64 were considered to be infected. The number of positive cases was 224 that is 23.5 percent of total received samples. Positive cases were found mostly in older animals [over 2 years] and in goats and rams perhaps because of butting. It has been suggested that the use of seroagglotination test may be helpful in detecting infected animals in contaminated area