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 characterization and phylogenetic study based on matrix gene of avian influenza viruses [H9N2] in Iran during 1998-2008]

Background: H9N2 avian influenza viruses [AIV] A have become panzootic in Eurasia over the last decade and have caused several human infections in Iran since 1998 and inactivated vaccine has been used in chickens to control the disease

Objectives: The purpose of this study was to analyze H9N2 viruses that have infected broiler in Tehran Province, Iran between 1998 and 2008 based on Matrix gene

Methods: The complete coding region of Matrix [M] gene from 8 of H9N2 subtype isolated from chicken flocks in Tehran Province during 1998-2007 was amplified and sequenced

Results: Sequence analysis and phylogenetic studies of H9N2 viruses on the basis of data of viruses in this study and other selected strains available in the GenBank were conducted and determined variations among these sequences at different levels. Sequence analysis revealed a large number of similar substitution mutations and close evolutionary relation among sequences of M gene. Phylogenetic analysis showed that all our isolates belonged to the G1-like sublineage. In this study, it was determined that Iran's isolates have been in two separate branches and have the most similarity with Pakistan, United Arab Emirate and occupied Palestine's isolates

Conclusions: The available evidence indicates that M genes of H9N2 circulating in Iran during the past years were not well conserved. Our finding emphasizes the importance of reinforcing AIV surveillance

Phylogenetic study based on the phosphoprotein gene of Iranian Newcastle disease viruses [NDV] isolates, 2010 -2012

Newcastle disease virus [NDV] is the causative agent of the Newcastle disease [ND], a highly contagious disease in birds that causes significant economic losses to the poultry industry worldwide. ND is endemic in Iran and outbreaks are reported regularly in commercial poultry flocks and different species of birds. The current study was carried out to characterize NDV based on phosphorprotein [P] gene from recent outbreaks in Iran, 2010-2012. The P gene fragment of NDV isolates of five chickens, 1 ostrich, and 1 Pigeon paramyxovirus-1 was obtained by RT-PCR and sequenced. Phylogenetic analysis of sequences revealed that chicken and ostrich NDV isolates were closely related and placed in the genotype VII and Pigeon Paramyxovirus-1 was located in the genotype V. This is the first report of Phosphoprotein gene sequences of NDV strains isolated in Iran. This study will help us to understand the epidemiology and molecular characteristics of Newcastle disease virus in Iran

Construction of a recombinant vector for site-directed mutagenesis in Salmonella typhimurium

Among all common techniques in site directed mutagenesis, Lambda Red recombinase system has been widely used to knock out chromosomal genes in bacteria. In this method, there is always the risk of DNA Linear digestion by host's restriction enzymes that leads to the low frequency of recombination. To overcome this, we constructed a recombinant vector to disrupt phoP gene in Salmonella typhimurium. The SOEing PCR method and restriction enzymes were used to construct the vector. The resulting plasmid, pTAAZ92, contains a Kanamycin cassette with two long homologous arms flanking of the phoP gene. After electrotransformation of the pTAAZ92 into the Salmonella typhimurium, the phoP gene is replaced by the Kanamycin cassette through homologous recombination. According to the high homology of the phoP gene in many of Salmonella species the pTAAZ92 can be used to disrupt the phoP gene in most of these species

[ molecular and clinical survey of feline herpesvirus-1 and feline calicivirus prevalence in non-vaccinated cats in Tehran, Iran]

Despite the widespread use of vaccines against feline herpesvirus type1 [FHV_1] and feline calicivirus [FCV], clinical signs are still seen in cat population. The aim of this study was to determine the prevalence of FHV-1 and FCV in non vaccinated clinically healthy cats and cats with URTD. Oropharyngeal and conjunctival swabs were taken from 16 cats with clinical signs of URTD and 26 clinically healthy cats. Reverse transcription polymerase chain reaction [RT-PCR] and polymerase chain reaction [PCR] were performed to diagnose FCV and FHV-1 infections. In cats with URTD, the prevalence rate of FCV [100%] was higher than FHV-1[43%] but in clinically normal cats the prevalence rate of both viruses was about 50%. Clinical signs in cats with FCV was more various than FHV. Also, the prevalence of both viruses co-infection was 30% and half of them showed clinical disease. The results indicated higher rate of both viruses infection especially FCV in domestic cats in Tehran in comparison with other regions. Stomatitis was seen in 50% of cats with URTD. In 50% of cats with corneal ulcers, contrary to our expectations, FCV was detected not FHV-1. It seems that new variants of calicivirus are more virulent and are able to damage other tissues like cornea and conjunctiva. FCV tends to produce more clinical signs than FHV. Also, infection with new variants of FCV and FHV-1 in healthy cats and cats with URTD is much higher than other regions of the world. Therefore, a revision of vaccines and vaccination program, especially for FCV has become a matter of necessity just like other countries

Distribution of virulence associated genes in isolated Escherichia coli from avian colibacillosis

Colibacillosis is one of the most prevalent diseases in the world that causes multimillion-dollar annual losses. In order to evaluate molecular epidemiology of some virulence associated factors in Escherichia coli, isolated from poultry, the presence of iut A, iss, hlyF, omp T, iro N, afa, sfa [S] and pap G [II] were investigated by multiplex PCR assay. Two hundred thirty four Escherichia coli isolated from avian colibacillosis [APEC] and fifty four fecal E. coli isolates from the feces of apparently healthy birds [AFEC] were investigated for presence of some virulence associated genes by two panel of multiplex PCR. Statistical analysis was performed using x[2] test, the p-value was

Development and ELISA-based detection of anti-M2e IgY antibodies using an encoding plasmid for M2e-Hsp70 c-terminal gene

The use of IgYs in a variety of methods in different areas of research, diagnostics, medical application and biotechnology should be considered widely. Development of antibodies against extra cellular domain of influenza M2 [M2e] protein in egg yolk of laying hens. A Fusion construct harboring C-terminal of bovine heat shock protein 70 [Hsp70] and influenza M2e coding genes was injected to laying hens. Serum and egg yolk antibodies were screened for the presence of anti-M2e antibodies by indirect enzyme-linked immunosorbent assay [ELISA]. Anti-M2e antibodies were detected in egg yolks and sera of injected hens from 13 and 7 days post injection [PI], with the peak titer detected on 41 and 35 days PI, respectively. Anti-M2e IgY titers could be an index for expression potential of pcDNA3.1-M2e-HspCterminal construct in laying hens. This construct could be considered as a promising tool in production of anti-M2e polyclonal, monospecific IgY antibodies. Such anti-M2e antibodies could be exploited for influenza diagnostic and therapeutic measures

[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