m. r .seyfi abad shapouri

Newcastle disease, as one of the most important diseases of the poultry industry, has many economic losses. The aim of this study was to investigate the role of Newcastle disease virus in respiratory syndromes of broiler flocks in Khuzestan and Isfahan provinces. For this purpose, tissue samples (trachea, lung, and cecal tonsils) were taken from ten dead birds of 20 broiler farms with respiratory symptoms in different regions of these two provinces. Samples were inoculated into 9 to 11 days old embryonated chicken eggs. The RT-PCR was performed to detect Newcastle disease virus on allantoic fluid. Out of 20 studied flocks, the virus was isolated and identified in nine flocks. The 330 base pair nucleotide sequence in the F gene virus cleavage site was examined in 9 isolates. The results of RT-PCR test showed that Newcastle disease virus was isolated from respiratory syndromes in 50% of broiler flocks in Khuzestan province and 40% of broiler flocks in Isfahan province. Of the 9 isolates, 3 were in genotype II and 6 were in genotype VIId of class II Newcastle disease virus, and isolates of both genotypes were present in both provinces. The variety of VIId isolates found in Khuzestan province was more than Isfahan province. Isolates belonging to genotype II in this study were similar to Pakistani, Chinese and Nigerian strains, and other isolates were similar to previous Iranian strains.

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.

Viral haemorrhagic septicaemia (VHS) is one of the most important viral diseases in rainbow trout that has caused great losses to the Iranian rainbow trout aquaculture industry in few recent years. In the early autumn of 2015, in order to identify the possible causes of loss, farmed fish were sampled from the epicenter where massive mortalities and economic losses had occurred. Three samples were collected from relevant tissues (i.e. liver, kidney, spleen, heart, and brain) of the rainbow trout with clinical signs of VHS disease. The samples were stored in 70% ethanol, Eagle’s minimum essential medium (EMEM) and 10% formalin to perform RT-PCR, cell-culture and histopathology examinations, respectively. A virus was isolated using fish cell lines inoculated with a homogenate of the internal organs. Sequencing analysis of truncated N gene confirmed the existence of VHS virus. RT-PCR and histopathology tests indicated the suitability and simplicity of the test as a paraclinical diagnostic method for VHS virus in the region. Haemorrhage, necrosis and other changes in histological sections of the diseased fish demonstrated pathological changes identical with the typical VHS pathology in target organ of the virus (i.e. heart, spleen, liver and kidney). To our knowledge, this is one the first studies that fully focused on the identification of VHSV from farmed trout in Iran by using histopathology examinations.

Canine parvovirus type 2 (CPV-2) is one of the most common viruses responsible for acute hemorrhagic enteritis in dogs. A rapid and accurate diagnosis of CPV-2 infection is especially important in kennels in order to isolate infected dogs. The aim of the present study was to compare two laboratory tests i. e.، Polymerase Change Reaction (PCR) and Immunochromatography assay (ICA) most commonly used for the diagnosis of canine parvovirus infection in companion dogs. Fecal samples were collected from fifty five dogs (50=hemorrhagic diarrheic and 5= healthy) between 2011 and 2012 in Ahvaz district، southwest of Iran. The studied dogs were divided into two age groups (<6 months، and>6 months)، four different breeds (Terriers، German shepherds، Doberman pinschers and Mixed) and based on environment into two groups (open and close) also. All samples were tested by ICA and PCR methods and the results were analyzed by using Kappa test، Mc Nemar and Chi-square analysis. ICA and PCR were able to detect CPV-2 antigen or nucleic acid in 33 and 50 of the hemorrhagic diarrheic samples، respectively. Samples of healthy dogs were negative by both tests. Although sensitivity of ICA compared with PCR method was determined to be 66% (PCR more sensitive than ICA)، nevertheless statistical analysis showed that the difference between two techniques were not significant (P>0. 05). Kappa test was obtained 0. 38 between two techniques. CBC showed that most infected dogs had leucopenia، lymphopenia and neutropenia also (82%; 41 out of 50 samples). Obtained results of this survey showed that accurate standardization of laboratory tests is required to provide veterinarian with an effective tool for a precise etiological diagnosis of hemorrhagic gastroenteritis due to CPV infection. Although Immunochromatography is a simple and quick method for screening of fecal samples of dogs suspected of CPV infection، but PCR is more sensitive and reliable than ICA. Moreover، the subtypes of the virus determined by PCR test after verifying parvovirus. In this test 48 samples were CPV-2b and another 2 samples were CPV-2a. Our results showed that CPV-2b was the predominant subtype.

The molecular detection of avian infectious bronchitis virus by use ofRTPCR and multiplex nested PCR in Fars province of Iran was investigated. Detection has been done on tracheal swab s:!mnles of 30 broiler tlocks in age of 7-8 weeks. Flocks were selected from nIU~' broilers rising regions orthe province. Detection was performed by primers specifie for Massachusett, 4/91 and 0274 serotypes. In this study 16 samples were positive for IBV 4/91 typeand 1 for Massachuset type. One swab sample showed a mixed infection of those serotypes. 0274 serotype has not been detected in this study. This is the first report of the presence and a prevalence of IBV type 4/91 in Fars province.