فهرست مطالب هادی کیوانفر

During the past two decades, the highly pathogenic avian influenza H5N1 virus has received considerable attention due to its zoonotic and mutative features. Purpose of the leading study is molecular and phylogenetic characteristics of hemagglutinin (HA) gene of H5N8 strain identified in Meighan wetland of Arak city, Markazi province were investigated. For this purpose, samples were inoculated with embryonic eggs of 14-10 days and after extraction of allantoic fluid and RNA extraction and PCR and sequencing of genes, phylogenetic trees were drawn by Mega7 program and molecular properties including Cleavage site, Glycosylation site, Antigenic site, Receptor Binding site and HA gene-related mutations were investigated. Based on the analysis of the amino acid sequence of the HA genes, the cleavage site of the gene includes the PLREKRRKR / GLF polybasic amino acid motif, which is a characteristic of highly pathogenic influenza viruses. The HA gene of two viruses had T156A, S123P, S133A mutations associated with the increased mammalian sialic acid binding. Phylogenetic analysis of the HA gene of the virus studied in this study indicated the classification of this virus in the 2.3.4.4 b Clade. It seems that the introduction of these H5N8 HPAI strains in Iran probably occurred through the West Asia-East African flyway by wild migratory aquatic birds.

The most common cause of milk production loss in cattle and water buffalo is bovine ephemeral fever (BEF). Previous cases have been reported in Iran's south regions, with a low mortality rate. As a result, studying BEFV and identifying ideal epitopes for further developing diagnostic paths is important.

To investigate BEFV N protein epitopes, we collected samples, extracted and sequenced DNA, and then used the ExPaSy translate method to deduce the amino acid sequence. Various immunoinformatics techniques were used to analyze physical/chemical properties, secondary structure of protein sequences, membrane topology, antigenic property, and 3D structure. BCPRED and the DiscoTope server, respectively, predicted linear and discontinuous epitopes of BEFV N protein. Finally, the PatchDock server was used to dock peptides and antibodies.

Three linear epitopes and sixteen discontinuous epitopic positions were discovered. Furthermore, molecular docking between epitopes and low-binding-energy antibodies revealed that they have easy access to the immune system.

In this study, bioinformatics techniques were used to predict epitopes of the BEFV N protein for further developing BEFV diagnostic paths. Furthermore, experimental validation is needed for these epitopes.

Peste des Petits Ruminunts (PPR) is an acute and contagious viral disease with rapid infection and transmission in sheep and goats. Vaccination of livestock is the best solution to prevent the spread and control of the disease. The aim of this study was to evaluate the immune response derived from vaccination with six formulations of inactivated PPR vaccine in goats and sheep using a completely randomized design with 2 × 2 factorial arrangement. The attenuated PPR virus was amplified in a highly manipulated suspension lymphoid cell having a high passage potential, and then inactivated using two inactivators (4 mM bromoethylene imine [BEI] or 3% hydrogen peroxide [H2O2]). Thereafter each inactivated virus was formulated using three different helpers (aluminum hydroxide, aluminum phosphate, and a 50% mixture of each of these two adjuvants). Iimmunogenicity and safety of the formulated candidate vaccines were evaluated by respective two doses of 0.1 and 1 (10-fold doses) ml of the vaccines administrated into 38 sheep and 28 goats. Each dose was subcutaneously administrated on 0 and 21 days of the experiment and post vaccination sera antibody titer against PPR virus was measured on days 0, 14, 21, 35, 42 and 60. The results showed that the mean antibody titer against PPR virus was not influenced neither by inactivator nor adjuvant up to 21- and 60-days post vaccination (P> 0.05). Also, no significant interaction was observed between different inactivators and adjuvants (P> 0.05). The appropriate antibody titers observed in different experimental groups indicated a good immunization of the formulated vaccines; however, their efficacy remained to be investigated using challenge test. Also, due to the lack of carcinogenicity, cheapness and environmental compatibility reported for H2O2, it is nominated as an inactivator in production inactivated PPR vaccine.

Lumpy skin disease (LSD) is a highly contagious disease of cattle and buffaloes that cause considerable economic losses to the livestock industry in endemic regions. Although the virus neutralization index (VNI) is the gold standard test for the detection of LSD antibodies, the use of this test in seroepidemiology studies and serosurveillance is laborious and time-consuming. Accordingly, for serodiagnosis of capripox diseases, ELISA assay has been developed, but the lack of information about its performance and efficiency is one of the limiting factors in the use of this test in seroepidemiological studies. In this study, the diagnosis agreement between a recently developed ELISA based on P32 recombinant protein and the VNI method, as a gold standard for serological diagnosis of Capripox virus, was evaluated using 95 sera samples including 25 negative control, 14 positive control, and 56 suspected cattle sera. Result showed that all the negative control sera assessed by VNI and ELISA had no antibody titers against LSD. All the 14 (100%) positive control sera had VNI values ≥1.5 and were considered as the positive sera; however, by ELISA method, 13 samples (93%) were diagnosed as the positive. Among the 56 samples collected from LSD-suspected cattle, 16 samples (29%) were detected as the positive by VNI, whereas 13 samples (23%) were detected as the positive sera by ELISA method. There was no statistically significant difference between the percentages of negative or positive samples assessed by ELISA or VNI method (P>0.05), associated with the Kappa index value of 0.71 showing the substantial agreement. These findings indicating an acceptable agreement and performance of the developed ELISA system based on P32 recombinant protein in comparison to VNI method for diagnosing antibody against LSD virus in cattle.

Newcastle disease virus has been investigated in several studies as an oncolytic virus to treat human tumors. However, precise mechanism of tumor cell killing by this virus is unknown. The aim of this study was isolation of Newcastle disease virus from industrial poultry and examination of its ability to induce expression of the extrinsic and the intrinsic pathways of apoptosis-related genes in human breast carcinoma cells. Newcastle disease virus was isolated from industrial poultry by culture in embryonated chicken eggs and then identified by haemagglutination test and RT-PCR technique. MCF-7 breast carcinoma cells were infected with the isolated virus in vitro. After RNA extraction and cDNA synthesis, gene expression levels of BAX, BAK, caspases-3, -8, and -9 were assessed in the virus-infected tumor cells using real-time RT-PCR technique. PCR data analysis showed that gene expression levels of BAX and caspase-9 were significantly higher in virus-infected tumor cells than that in control tumor cells. Gene expression levels of BAK and caspases-3 and -8 were not significantly altered in virus-infected tumor cells in comparison with control tumor cells. These results suggest that Newcastle disease virus isolated from industrial poultry can trigger the intrinsic pathway of apoptosis in MCF-7 tumor cells.

Rural poultry like commercial poultries are susceptible to most of infectious diseases. In addition, by increasing the density of poultry farming, the probability of disease occurrence has been increased. Among the most important diseases, Newcastle disease has most of importance. Newcastle disease is endemic in Iran, and causes incidence of outbreaks among commercial and rural poultries, every year. The present study is conducted with the objective of figuring out the prevalence status and virus circulation among rural poultries of Northern provinces of Iran. In the study, 70 villages in 3 provinces (20 villages in Mazandaran, 20 villages in Golestan and 30 villages in Gilan Province) and a total of 1374 birds (600 birds in Mazandaran, 400 birds in Golestan, 374 birds in Gilan province) were sampled. A village considered as epidemiological unit. In the study, birds of 67 villages (96%) were found positive ( presence of antibodies against NDV) including Golestan Province, 28 villages (93.3%), Mazandaran Province, 19 villages (95%) and 20 villages of Gilan province (100%) Moreover, out of 1374 birds, 616 (45%) of them were seropositive against NDV. According to the results of this study, the rate of titer is very high in both levels of villages and level of birds. Such high rate of titer is indicative of continuous exposures of rural poultry of the mentioned provinces to Newcastle virus and high virus circulation rate of these viruses in the studied provinces.