Development of real-time reverse transcriptase-polymerase chain reaction (rt RT-PCR) targeting four genes of peste des petits ruminants virus

In this study, one-step real-time quantitative reverse transcription PCR (qRT-PCR) was developed for the first time and evaluated for detection of peste des petits ruminants virus (PPRV) in field samples obtained from distinct geographical areas of Turkey. Primers and probes targeting four PPRV genes (namely L, M, N and P) were designed based on full genome sequence (AJ849636) of the local isolate (Tu00). The detection limits of the assays were found to be 7 copies/mu L RNA for L, 6 copies/mu L RNA for P, 10 copies/mu L RNA for M and 700 copies/mu L RNA for N, respectively. Besides, the detection ratio for PPRV in 45 field samples was 100% (45) for L, 88.9% (40) for M, 77.8% (35) for P and 22.3% (10) for N, respectively. In conclution, one-step real-time quantitative reverse transcription PCR (qRT-PCR) assay reported here for L gene design provides rapid, specific and sensitive detection of PPRV in tissue samples obtained from field cases

The first detection and molecular characterization of porcine reproductive and respiratory syndrome virus (PRRSV) in Turkey

Porcine Reproductive and Respiratory Syndrome (PRRS) is an important disease that causes severe economic losses in pig industry. PRRSV has two genotypes named as a European (EU) and North American (US). PRRSV appears globally a variety of countries including Canada, USA, South Korea, Germany, Spain, UK, Denmark and Greece. Furthermore, US genotype has been detected serologically in France, Germany and USA in wild boars. So far, no comprehensive information has been generated in Turkey in terms of presence and/or epidemiology of the disease. The aim of this study was to determine presence of PRRSV at initial step and subsequently provide information on possible genotypes that might have occurred in domestic pig and wild boars. The study was carried out on 86 nasal swaps from two different farms and 12 lungs tissue samples from wild boars. A total of 71.4% of samples were found PRRSV- positive by one-step RT-PCR. Prevalence of the virus in Farms 1 and 2 and wild boars was determined as 76.9%, 61.9% and 58%, respectively. Nucleotide sequence analyses performed on ORF 7 of the genome showed that 96.5% and 98.2%, nucleotide homologies in Izmir and Mersin, respectively. In addition, phylogenetic analysis showed that all Turkish PPRSVs was located in the lineage of US like PRRS viruses. This is the first report of PRRSV infection between domestic pigs and wild boars in Turkey

Phylogenetic analysis of black queen cell virus and deformed wing virus in honeybee colonies infected by mites in Van, Eastern Turkey

This study aimed to determine the presence and prevalence of viral and parasitic infections causing high rates of colony loss in honey bee colonies in Van province, eastern Turkey. Twenty-six different apiaries were collected from five counties in Van province. These samples were tested by Reverse-Transcriptase PCR (RT-PCR) for acute bee paralysis virus (ABPV), chronic bee paralysis virus (CBPV), black queen cell virus (BQCV) and deformed wing virus (DWV). Selected positives were sequenced, phylogenetically analyzed and investigated in terms of Varroa. DWV and BQCV were identified in 69.23% (18/26) and 88.46% (23/26) of the bees respectively whereas ABPV and CBPV were not detected in the sampled apiaries. Results of the phylogenetic analysis of DWV and BQCV sequences showed 94-100% similarity to DWV and BQCV isolates obtained from Genbank. Prevalence of varroasis was 89% (23/26) in Van. The obtained samples were identified as Varroa destructor by morphological investigation. The study showed that viral and parasitic agents commonly infect honeybees in Van province, with high prevalence rates for BQCV and DWV. There was also a high degree of conservation of DWV and BQCV sequences distinct from DWV and BQCV isolates from other geographical regions. These findings, including current prevalence and phylogenetic analysis data for DWV, BQCV and varroazis in honeybees, are useful for future studies

Pathology and Phylogenetic Analysis of Capripoxvirus in Naturally Infected Sheep Sheeppox Virus

Sheeppox and goatpox is a contagious viral disease of sheep and goats characterized by fever, generalized papules or nodules in skin and mucosal surface. DNA virus belonging to the genus capripoxvirus. The disease with high mortality and morbidity causes significant economic losses in small ruminants. In this study, 20 tissues from 8 sheep which were considered having natural sheeppox based on the macroscopic and histopathologic evaluation were further investigated by immunohistochemical (iNOS, SP-A, HSP-70), and Polymerase Chain Reaction (PCR) method. Papules, typically pox lesions, noticeable on the skin surfaces were examined macroscopically. Immunohistochemically; iNOS and SP-A were most intensely stained whereas HSP-70 was low stained. PCR method was used for the detection of A29L gene of capripoxvirus. Positive samples obtained from sheep were used for molecular characterization. A phylogenetic analysis was performed using sequence of the partial A29L gene and by comparing with reference sheeppox viruses isolates obtained from Gene Bank. The results of the sequence analysis were similar among themselves, they were found different (99-100% identity) from the other sheeppox viruses around the world. This study provides firstly phylogenetic analysis of sheeppoxviruses from Van province in Turkey. (C) 2016 PVJ. All rights reserve

Molecular detection of Nosema spp. and black queen-cell virus in honeybees in Van Province, Turkey

This study was planned to determine the prevalences of the Nosema spp. and the black queen-cell virus (BQCV) among honeybees (Apis mellifera) raised in the province of Van by PCR and to determine the molecular characteristics of the determined isolates. A total of 260 adult worker bees from 26 colonies at 5 apiary locations belonging to the province of Van in April and May 2015 were collected for this reason. Samples were examined microscopically. In the case of positivity, spore identification was done by multiplex PCR. Reverse transcription/PCR analysis (RT/PCR) was carried out for the BQCV analysis. At the end of the microscopic examination, Nosema spp. spores were detected in 8 out of 26 colonies (32.5%). The result of multiplex-PCR revealed Nosema ceranae positivity in all of the samples, but no Nosema apis was determined. As a result of the RT/PCR tests of the samples BQCV was detected in 23 (88.5%) of the total 26 colonies. This study is the first to investigate Nosema spp. and BQCV with the PCR technique in bees raised in the province of Van

Molecular and restriction fragment length polymorphism analysis of canine parvovirus 2 (CPV-2) in dogs in southeast Anatolia, Turkey

Canine parvovirus-2 (CPV-2) is the aetiological agent of an infectious viral disease of dogs, characterised by diarrhoea and vomiting. Mutations of the CPV-2 genome have generated new variants circulating worldwide. This article reports the molecular analysis of CPV-2 variants collected in the dog population in southeast Anatolia, Turkey. Twenty blood samples previously taken for the laboratory diagnosis of dogs with suspected parvovirus were screened for CPV-2 by polymerase chain reaction (PCR). Of the 20 samples, 18 tested positive for CPV-2. Partial VP2 gene sequencing and restriction fragment length polymorphism (RFLP) analysis revealed CPV-2a (n = 1), CPV-2b (n = 16) and CPV-2c (n = 1) variants. Phylogenetic analysis based on the partial length VP2 gene showed that CPV-2b (n = 15) variants showed sequences clustering separately in the phylogenetic tree. The CPV-2c sample was phylogenetically related to Chinese strains and Indonesia strain, whereas the CPV-2a sample was phylogenetically related to the Portuguese strain. These results, which are the first to demonstrate the presence of CPV-2c in the dog population of southeast Anatolia, Turkey, indicate that CPV-2a/2b/2c variants co-exist in Turkey's dog population

Relationship Between Crimean-Congo Hemorrhagic Fever Virus Strains Circulating in Iran and Turkey: Possibilities for Transborder Transmission

Crimean-Congo hemorrhagic fever (CCHF) is an important zoonotic viral disease that is asymptomatic in infected livestock, but poses a serious threat to humans. The high fatality rate may be due to phylogenetic variations in the virus, transmission routes, and a lack of an efficient surveillance system for the disease. The geographical features of the eastern and southeastern borders of Turkey may facilitate transmission of viruses between countries of the region. Therefore in this study we focused on the genetic relationship between Turkish and Iranian CCHF viruses based on their S-segment sequences. The research was performed on a total of 104 blood samples from small ruminants reared in southwest Iran. The results of phylogenetic analysis showed that Iranian CCHF virus isolates were closely related to human-originating Turkish Group II viruses from a European lineage reported previously

Molecular epidemiology of Crimean-Congo hemorrhagic fever virus in Turkey: Occurrence of local topotype

The goal of this study was to investigate the molecular epidemiology of Crimean-Congo hemorrhagic fever virus (CCHFV) in Turkey. The study was performed on a total of 48 confirmed human CCHF cases from 2006 to 2008. The majority of the CCHF viral strains in Turkey were found to belong to the European lineage. Local CCHF viral strains are grouped into two main clusters, which can be further divided into two sub-groups. We also identified an AP92-like virus causing clinical disease in Corum (a mid-Anatolian province). Phylogenetic analysis revealed that the most recent CCHFV infections were caused by intrinsic (or native) CCHF viral strains, which we identified as the local topotype. Comparison of deduced amino acid sequences of S-segment RNAs indicated that the local topotype was derived from viruses of previous years, most likely by a low rate recombination. No genetic differences, based on S- and M-segment RNA sequences, were found between human and tick viral isolates. This data suggest that replication of CCHFV in the tick vector, whether Rhiphicephalus spp. or Hyalomma spp., has no effect on the viral genomic structure. (C) 2010 Elsevier B.V. All rights reserved

A Snapshot Avian Surveillance Reveals West Nile Virus and Evidence of Wild Birds Participating in Toscana Virus Circulation

Introduction: Birds are involved in the epidemiology of several vector-borne viruses, as amplification hosts for viruses, dissemination vehicles for the vectors, and sources of emerging strains in cross-species transmission. Turkey provides diverse habitats for a variety of wild birds and is located along major bird migration routes. This study was undertaken to provide a cross-sectional screening of avian specimens for a spectrum of vector-borne viruses