Search

Background: Rotavirus (RV) is one of the most important causes of diarrhea in the calf and human neonates. Rotaviruses are divided into nine different serogroups, of which group A is more important compared to other groups. Objective: This study was performed because of the lack of information about the importance and prevalence of bovine rotaviruses B (RVB) and C (RVC) and human genotypes of rotavirus A (RVA) in the bovine population in Iran. Phylogenetic analysis of VP6 of bovine RVA was the second aim of the present study. Materials and Methods: A total of 581 stool specimens were collected from diarrheic calves of 14 provinces and were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and 485 of them were investigated by PAGE electrophoresis to determine the frequency of three rotaviruses A (RVA), B (RVB), and C (RVC). The presence of human G and P genotypes in Iranian bovine population was also evaluated using semi-nested multiplex RT-PCR. Results: RVA was detected by RT-PCR (VP6 gene detection) in 16.2% (94/581) and by PAGE in 22.16% (108/485) and no positive cases of RVB and RVC were confirmed by polyacrylamide gel electrophoresis (PAGE). This study showed that non-A RV groups (B and C) have little role in calf diarrhea in Iran. The results of the phylogenetic study of VP6 sequences of rotaviruses A identified in this study showed that they all belonged to genotype I2 and were classified into three different branches. Specimen isolated in Zanjan showed the highest difference (maximum identity of 94%) with other sequences and clustered along with the Japanese strain, R22. Human G and P genotypes were not found in the studied samples. Conclusion: The results showed that non-A rotaviruses and human genotypes of RVA are of little importance in calf rotavirus diarrhea in Iran. Also, there is the first phylogenetic study of rotavirus A VP6 protein in Iran.

Panleukopenia is a viral enteritis of high mortality rate in Feline population. It is caused by a small single-stranded DNA virus, belonging to the Family Parvoviridae, called Feline Panleukopenia Virus (FPV). Parvoviruses are capable of adapting to new hosts and can simply mutate like RNA viruses, resulting in the emergence of different strains with antigenic changes. Unlike the original Canine Parvovirus (CPV) type 2, the antigenic variants (CPV-2a, CPV-2b, CPV-2c) earned the ability to replicate in cat hosts either with clinical signs or without any symptoms. Cats may be able to shed Parvoviruses for a long time and can serve as an important reservoir. Co-infection with FPV and CPV-2 variants has been also reported. These characteristics make the cat an important host for Parvoviruses. Since no molecular study has reported on Parvoviruses infected cats in Iran, a decision was made to study these strains of Parvovirus. Twenty-eight fecal samples were collected from kittens under 6 months of age, both male and female and different breeds which had leukemia. PCR was performed on each sample and 26 samples tested positive for Parvoviruses. Randomly, the PCR products of eight samples were sequenced and their phylogenetic tree was generated. Analysis of the results revealed that all eight samples are FPV and it appears that FPV is more clonal than the CPV-2 variants in Iran. Amino acid residue 232 from the VP2 gene of FPVs has been changed from isoleucine to valine, which has only been reported in CPV-2 and its variants. It seems that some FPVs of Iran are changing to the CPV-2 variants. Hence, it is expected that in the future, the CPV-2 variants will be observed in cats.

Mammalian T7 polymerase-based cytoplasmic expression systems are common tool for molecular studies. The majority of these systems include the internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV). To carry out a cap-independent translation process, this type of IRES might require the expression of an extensive array of host factors, what is a disadvantage. Other IRESes might be less dependent on the host cell factors, but their biology is characterized to a lesser degree. Here, we compare the translational efficiencies of bovine viral diarrhea virus (BVDV) IRES with that of ECMV. Both IRESes were tested in reporter vectors containing the T7 promoter, an IRES of choice and the coding sequence of the enhanced green fluorescent protein (EGFP). To provide for the expression of T7 RNA polymerase, the corresponding gene was isolated from Escherichia coli and inserted into pCDNA3.1-Hygro(+). After co-transfection of the T7 RNA polymerase encoding vector with either of the two IRES-containing reporter vectors into T7 baby hamster kidney (T7-BHK), human embryonic kidney (HEK) 293T, chinese hamster ovary (CHO) and HeLa cells, the translational efficiency of the reporter construct was studied by fluorescence microscopy and flow cytometry. In T7-BHK, HEK 293T and HeLa cells the translational efficiency of BVDV IRES was two to three times higher than that of EMCV IRES. In CHO cells, BVDV IRES and EMCV IRES were equally efficient. An analysis of the secondary structure of respective mRNAs showed that their ΔG values were–544.00 and–469.40 kcal/mol for EMCV IRES and BVDV IRES harboring molecules, respectively. As EMCV IRES-containing mRNA is more stable, it is evident that other, still unidentified factors should be held responsible for the enhanced translational efficiency of BDVD IRES. Taken together, our results indicate the potential of BVDV IRES as a replacement for EMCV IRES, which is now commonly used for T7 polymerase driven cytoplasmic expression of genes of interest or virus cDNA rescue experiments.

Purpose. Bovine leukemia virus (BLV) infects cattle worldwide, imposing an economic impact on the dairy cattle industry. The purpose of this study was to evaluate the molecular epidemiology of BLV in Iran. Methodology. Blood samples taken from 280 cows aged over 2 years old from 13 provinces of Iran were used for leukocyte count and blocking ELISA. Genomic DNA was extracted from the peripheral blood leukocytes of BLV-infected samples and fetal lamb kidney cells to perform PCR of partial env, rex and tax genes and long-terminal-repeat region. The PCR products were sequenced, the phylogenetic tree of each gene was constructed, and nucleotide and amino acid sequence pair distances were calculated. Results. The frequency of BLV infection was 32.8 % among animals and was 80 % among provinces. In BLV seropositive animals, the rate of persistent lymphocytosis was 36.9 %. The constructed phylogenetic trees showed the presence of two BLV genotypes (1 and 4) in Iranian strains. As previous studies, our results showed that the env gene was more variable than previously thought, the Rex protein could withstand more amino acid changes compared to the Tax protein, and no significant differences were observed in average changes of the nucleotide of these genes between clinical stages. Conclusions. Our data indicates an increase in the frequency of this infection in Iran. This is the first study report of the presence of BLV genotype 4 in Iranian farms. These findings may have an important role in the control and prevention of BLV infection in Iran and other countries.

Background Breast cancer is one of the most common cancers in the world particularly among Iranian women. Bovine leukemia virus (BLV) is an enzootic, exogenous, and oncogenic retrovirus that causes B-cell leukosis in 1–5% of infected cattle. The current study aimed at evaluating the correlation between BLV infection and breast cancer in an Iranian population. Materials and techniques A total of 400 samples including 200 breast cancer-suspected tissue samples and 200 blood samples of women without breast cancer, were collected from July 2017 to October 2018 from women referred to two general hospitals in Qom Province, Iran. The nested PCR technique was performed to determine the presence of tax and gag gene of BLV in the collected samples. Results Out of 200 breast cancer-suspected tissue samples, 172 samples were malignant in terms of pathology. Other samples were reported as non-malignant and non-tumor. Based on nested PCR technique, tax and gag genes of BLV were detected in 30% and 8% of breast cancer-suspected tissue samples, respectively. The frequency of BLV in blood samples collected from women without breast cancer was 16.5% (33/200). Conclusion It seems that human breast cancer and BLV infection in cattle could be associated using nested PCR technique.

Purpose: The direct transmission of avian influenza viruses to human and increasing drug resisted strains posing new threats for public health. Therefore, development of efficient vaccines is needed to generate protective and persistent immunity to the viruses. Methods: Three motifs of Mx protein sequence in human, mouse and poultry located in interferon induced (GTP ase) domain were candidate as biologic adjuvant for enhancing the immune responses against influenza virus. Chimera proteins composed with the conserved HA2 subunit of influenza virus and the Mx motifs named HA2/Mx were modeled and evaluated by in silico analysis includes bioinformatics algorithms in order to explore biological characteristics of these peptides. Results: Amongst the predicted models, HA2/Mx1 peptide showed the better results following protein structures prediction, antigenic epitopes determination and model quality evaluation. Comparative homology modeling was performed with Swiss Model and the model was validated using ProSA. Epitope predictions revealed the construct could induce both B and T cell epitopes that expect a high immune response. Conclusion: Taken together, these data indicate that the HA2/Mx1 chimera peptide can be potentiated for developing an adjuvant-fused influenza vaccine capable of stimulating effective immune response.

The most important causal agent of border disease (BD), border disease virus (BDV), is a Pestivirus in the family Flaviviridae. In attention to the lack of molecular documented data on the prevalence of BDV in Iran, especially in East Azerbaijan Province, northwest of Iran, this research was designed to determine the prevalence of BDV in suspected ewes that have an antecedent of reproductive failure. Eighty-eight suspected ewes in the breed of Ghezel, Mogan, and mix (Ghezel × Mogan) in five various districts of East Azerbaijan Province (Azershahr, Lyqvan, Osku, Sofian, and Varzqan) were selected. Razi bovine kidney (RBK) cell lines were grown, and the buffy coat layer was separated and inoculated to the monolayer cell culture. All cell cultures were inspected for cytopathic effects after 5–7 days. BDV genomes were being checked using nested reverse transcription polymerase chain reaction method. Ten samples (10/88, 11.36 %) were positive for the genome of BDV. The prevalence of BDV infection was found to vary between different districts. The highest percentage of infected sheep was in animals at the 3–4.5-year-old range (15.38 %). There was no statistically significant difference in the prevalence among different age categories. The highest BDV infection rate was observed in full blood ewes in Mogan and Ghezel. Cytopathic effects of the virus in the culture on RBK cell line were not observed at all. This study proved that the infection of BDV is present in sheep in the East Azerbaijan, northwest of Iran, and cause some reproductive failure in the flocks. The survey is the first molecular evidence of the occurrence of the disease in the country.

H9N2 avian influenza viruses (AIVs) have been recorded in Eurasian for several years. Since 2004-2005, the disease has become endemic in Iraq, causing serious economic losses in the poultry industry. The hemagglutinin (HA) and neuraminidase (NA), two out of eight protein-coding genes, play an important role during the early stage of infection and hinder virus assembling. Little is known about the genetic information of the H9N2 viruses currently circulating in Iraq; thus, gene sequences of six AIVS of the H9N2 subtype have been detected and analyzed in the period of 2014-2015 from different outbreaks of broiler flocks in five provinces situated in the middle and southern parts of Iraq. Genetic comparison of the partial sequences of HA gene indicated that all Iraqi viruses are related to each other and could be divided into two subgroups. Viruses of the first and the second subgroups demonstrated a high similar identity with Pakistani and Iranian viruses, respectively. The nucleotide sequences of the NA protein of the all studied Iraqi viruses were very similar (95.2-100% identity), and shared high nucleotide sequence identity with Iranian, Pakistani, and Lebanese strains. All six recent viruses possessed histidine, alanine, and leucine at positions 183, 190, and 226, respectively, which are the key residues in receptor-binding sites. The Iraqi viruses were closely related to viruses of G1-like lineage isolated from poultry flocks of Iran and Pakistan, suggesting that possible epidemiological links could be derived from a common origin. Further investigations are required and should include the viral isolation and full-length molecular characterization of H9N2 AIVs in this area.

Bovine viral diarrhea virus (BVDV) is an economically important animal pathogen affecting cattle. Despite the use of vaccination, test and slaughter practices, BVD remains a serious problem of cattle breeding. This study was conducted in order to develop a cell line that expresses some of BVDV sub-replicons. BVDV-NADL NS3 and 5’UTR were cloned in pWPI-linker B lentiviral plasmid at the upstream of EGFP gene. Consequently, lentiviral vectors containing BVDV-NS3 and BVDV-5’UTR were produced by using the second-generation lentiviral packaging system. By these lentivectors, MDBK cells expressing BVDV-5’UTR and BVDV-NS3 partial fragments were prepared. The efficiency of the infection was evaluated by fluorescence microscopy, western blotting, and RT-PCR. The results indicated that the development of MDBK cell line expressing these transgenes provides a very sensitive antiviral drug screening system for anti-bovine viral diarrhea virus (BVDV) therapies.

Upper respiratory tract diseases (URTD) are common clinical problem in cats worldwide. Feline calicivirus (FCV) and feline herpesvirus type 1 (FHV-1) are the main primary pathogens. Feline immunodeficiency virus (FIV) and Feline leukemia virus (FeLV) are also among the most common infectious diseases of cats which suppress the immunity. Oropharyngeal and conjunctival swabs and blood samples were taken from 16 cats with clinical signs of URTD and 26 clinically healthy cats. PCR and RT-PCR were used to detect FHV/FIV or FCV/FeLV infections, respectively. Feline calicivirus was detected in all cats with URTD and 87.00% and 93.00% of them were positive for FIV and FeLV, respectively. Feline herpesvirus rate of infection was 43.00% in sick cats. In clinically normal cats, prevalence rates of FCV and FHV were about 50.00%, but FIV and FeLV rates (42.00% and 65.00% respectively) were higher compared to other studies. Stomatitis was observed in 50.00% of cats with URTD. The main causative agent of corneal ulcers is FHV-1, but in 50.00% of cats with corneal ulcers, FCV was detected alone. It seems new variants of Caliciviruses are the main causative agents to attack uncommon tissues like cornea, although retroviral infections may be in the background of these various signs. The high retroviral prevalence may be due to existence of large population of stray cats. This is the first molecular study of FeLV and FCV in Iran and seems that FCV and FHV prevalence rates in FIV or FeLV infected cats is more than other non-infected ones.