The first serological detection of West Nile virus infection among residents living in northern Turkey.

BACKGROUND & OBJECTIVES: West Nile virus (WNV) is transmitted by a mosquito-borne virus whose natural reservoir is birds. Humans and horses are considered accidental hosts. Even if the vast majority of WNV infections in humans have asymptomatic or mild disease settings, serious neurological disorders with lethal outcomes can also be observed in around 1% of the cases. We aimed to serologically investigate the presence of WNV in humans living in Black sea of Turkey, and to obtain epidemiological data that will contribute to the implementation of public health policies to control and prevent potentially other life-threatening arboviral infections. METHODS: In the current study, a total of 416 human sera were collected from native patients of Samsun and its boroughs attending Samsun Training and Research Hospital; these sera were tested for WNV with pooling method, using anti-IgM and IgG ELISA commercial kits. All pools that were found positive for both IgM and IgG were individually retested for the detection of positive WNV sera. After that, all positive samples were tested using real-time PCR to detect the presence of WNV-RNA particles. RESULTS: Total seropositivity rates of WNV in terms of IgM and IgG were found as 0.96% and 0.72%, respectively. No presence of WNV-RNA could be detected in positive samples. INTERPRETATION & CONCLUSION: According to the data, further studies should be conducted to better understand the epidemiological dynamics of WNV in Turkey. It is recommended that other antigenically related flaviviruses which can give cross-reaction with WNV should also be investigated.

First isolation and molecular characterization of canine parvovirus-type 2b (CPV-2b) from red foxes (Vulpes vulpes) living in the wild habitat of Turkey.

BACKGROUND: The canine parvovirus, with its many variants, is responsible for a pivotal and common viral infection affecting millions of dogs and other carnivore species worldwide, particularly the wild ones, which are considered as the main reservoir hosts. To that end, this study investigated the presence of canine parvovirus (CPV) in red foxes (Vulpes vulpes) living in wild habitats of several regions of Turkey. METHODS: We randomly collected 630 archival fox stool specimens from rural areas of 22 provinces and used real-time PCR to detect CPV. RESULTS: Two of the 630 (0.3%) stool samples were positive for CPV-DNA, named Tr-Fox/128(Aydin) and Tr-Fox/159(Manisa). We attempted to isolate the virus in a MDCK cell line, and cytopathic effects were observed four days post-inoculation. Three regions corresponding to the CPV capsid protein VP2 gene from extracted DNA of positive samples were amplified by conventional PCR, and the products were visualised, purified, and Sanger sequenced. Three overlapping DNA raw sequence fragments, were read, assembled, and aligned to obtain approximately 1.5 kb-long regions that cover most of the VP2 gene, then deposited in GenBank. After comparing the isolates with parvovirus sequences data of domestic and wild carnivores by BLAST processing, our isolates' similarity rate with each other was 99.40%, with base differences in 9 nucleotide positions. They were classified as 2b variant closely related to isolates from dogs in Turkey, Egypt, Iraq, Italy, Thailand, and China. CONCLUSION: This study presents evidence of interspecies transmission of CPV, of which there are no reports on prevalence in wildlife carnivores of our country. Identification of CPV in red foxes threatens local and hunting dogs, which may contract the infection or disseminate it to other wild animal species or vice-versa.

A one-year extensive molecular survey on SARS-CoV-2 in companion animals of Turkey shows a lack of evidence for viral circulation in pet dogs and cats.

Current evidence have now demonstrated that SARS-CoV-2 infects a wide array of mammalian animals; however, the full range of hosts and the viral circulation in companion animals remains to be clarified. In this context, as no such evidenced cases have been reported from Turkey, we aimed to screen for SARS-CoV-2 nucleic acid in housed dogs and cats clinically evaluated for respiratory symptoms and reared in different locations of Samsun province in the black sea region of Turkey from July 2020 to July 2021. Nasal swabs were collected from a total of 415 pets (65 cats and 350 dogs) aged between 1 and 9 years old. All the specimens were tested for SARS-CoV-2 RNA presence by real-time RT-PCR targeting two genomic regions of SARS-CoV-2, but none showed positive results. Our findings suggest that SARS-CoV-2 does not circulate in local pets and is not responsible for respiratory symptoms. However, further comprehensive molecular and serological surveys are required to have a better picture of the zoonotic, reverse zoonotic and pathogenic consequences of the ongoing COVID-19 pandemic in Turkey.

The Border Disease virus (BDV) prevalence and genetic typing in ruminant flocks in Turkey.

This study aims to update current data regarding Border Disease in sheep and goats, determine the first prevalence of BDV in cattle and identify its circulated genotype in Turkey. For this purpose, 100 sheep, 20 goats and 193 cattle aborted fetuses sent for diagnosis to Samsun Veterinary Control Institute between 2015 and 2017 were analyzed in terms of pestivirus by Ag­ELISA, BDV by Real­Time test (RT­PCR) and Conventional RT­PCR test. The rate of pestivirus positive animals was found at 50.26% (97/193) in cattle, 58% (58/100) in sheep and 55% (11/20) in goats by the pestivirus Ag­ELISA test. Total of 58 Ag­ELISA positive sheep were tested by Real­Time RT­PCR and conventional RT­PCR tests. End of the tests, one sheep sample (1.72%) was found BDV positive by Real­Time RT­PCR test and three sheep (5.17%) and one cattle (1.03%) samples were detected as BDV positive by conventional RT­PCR test. BDV positivity was not detected in goats in this research. All samples that were found positive by conventional RT­PCR test and Real­Time RT­PCR test were genotyped by phylogenetic sequence analysis, and obtained results showed that BDV­3 and BDV­7 genotypes of BDV in sheep and BVDV­1 genotype in cattle circulated in the investigated area. The sequence analysis results revealed that conventional RT­PCR and Real­Time RT­PCR tests detected genotype BDV­3, while genotype BDV­7 was only detected by conventional RT­PCR test in sheep abortion materials. Additionally, it was found that one bovine specimen was BDV positive by conventional PCR, but the same sample was identified as BVDV­1 at sequence analysis. The obtained data of this study showed that new probes should be designed using our local strains for BDV diagnosis by Real­Time RT­PCR assay, and cattle must be sampled for BDV screening, and PCR tests results should always be confirmed by sequence analysis.

Serological study on the presence of some alpha-herpesviruses in goats of northern Anatolia, Turkey.

The aim of this study was to investigate the presence of caprine herpes virus-1 (CpHV-1) and bovine herpes virus-1 (BoHV-1) in 269 goat sera collected from small-scale family farms located in six provinces within the Black Sea region of northern Turkey. The overall seropositivity for alpha-herpesvirus in the native goats was found as 19.33% using BoHV-1 glycoprotein B (gB)-blocking enzyme-linked immunosorbent assay (ELISA). Additionally, the seroprevalence of BoHV-1 was determined in 5.20% of the goats using virus neutralization test. To distinguish between CpHV-1 and BoHV-1, the combinations of gB/gE-blocking ELISA tests were performed. Of tested samples, 15.24% were CpHV-1 seropositive; whereas, 4.09% were BoHV-1 seropositive. The results indicated that CpHV-1 is in circulation among local goats of northern Turkey. Considering the close relationship between BoHV-1 and CpHV-1, the transmission of BoHV-1 via goats may also be one of the predisposing factors involving in the spread of virus among the surrounding cattle.

Sentinel serosurveillance of backyard hens proved West Nile virus circulation in the western provinces of Turkey.

West Nile virus (WNV) is a mosquito-borne virus of a re-emergence importance with a wide range of vertebrate hosts. Granted, it causes asymptomatic infection, but fatal cases and neurologic disorders were also recorded, especially in humans, horses and some exposed birds. The virus is globally spread and birds are considered an amplifying and reservoir host of WNV, helping to spread the disease due to their close contact with main hosts. In this study, we aimed to detect the presence of antibodies against WNV in backyard hens that were reared in the western Anatolian part of Turkey. A total of 480 chicken sera were randomly collected from six provinces in the west of Turkey (Mugla, Izmir, Aydin, Afyonkarahisar, Kutahya and Manisa) with 80 samples from each province (40 in spring and 40 in fall seasons). They were tested by using a competitive ELISA method to identify the specific avian antibodies of IgG that produced against the WNV envelope proteins (pr-E). Twelve of 480 (2.5%) sera were found seropositive, three of these positive sera were detected from the Izmir province (3.75%) collected in the spring session and the other nine positive sera were detected from the Mugla province (11.25%) collected in the fall session. Both of these provinces are located seaside and have suitable climate conditions for vectors of infection. The results indicated that WNV infection is in circulation in these provinces, and that may put the other susceptible vertebrates under risk of infection.

A serosurvey for bovine respirovirus 3 in Turkish domestic ruminants: The first comparison study of A and C genotypes.

Bovine parainfluenza virus-3 (BPIV-3), also known as bovine respirovirus 3, causes serious respiratory infection in ungulates, often involving other pathogens, such as viruses, bacteria and mycoplasmas. In this study, we evaluated antibody titers against virus genotypes A (BPIV-3a) and C (BPIV-3c). We conducted a serological survey and comparison analysis of archived serum samples from small and large ruminants reared in four Turkish provinces. A total of 1,307 samples, consisting of sheep (n = 444), cattle (n = 402), water buffalo (n = 261) and goat (n = 200) sera, were randomly selected from stock samples collected between 2015 and 2019 and screened by standard virus neutralisation assay. We found that 49.9% (653/1307) of all samples were positive for neutralising antibody titers. Goats had the highest titer, with total seropositivity of 63% (126/200), followed in descending order by cattle, sheep and water buffalo at 56.2% (226/402), 32.2% (143/444) and 26% (68/261) total seropositivity, respectively. BPIV-3c had the highest neutralising antibody rate at 34.3% (448/1307), whereas BPIV-3a had a 24.3% (317/1307) seropositivity rate. Neutralising antibody titers for positive samples ranged between 1/4 and 1/512 per the SN50 test. Seropositivity rates ranged from a low of 8.9% to a high of 18.3%. Our study was the first to compare antibody seroprevalence for two BPIV-3 genotypes in small and large domestic ruminants, which were shown to be more commonly exposed to BPIV-3c than BPIV-3a. This finding could have significant implications as current vaccines mainly use the BPIV-3a genotype. Further research can determine if current vaccines protect against different BPIV-3 virus genotypes.

Marek's disease virus in vaccinated poultry flocks in Turkey: its first isolation with molecular characterization.

Marek's disease (MD) is an important disease of avian species and a potential threat to the poultry industry worldwide. In this study, 16 dead commercial chickens from flocks with suspected MD were necropsied immediately after death. Pathological findings were compatible with MD, and gallid alphaherpesvirus 2 was identified in PCR of spleen samples. Virus isolation was performed in primary cell culture, and partial sequencing of the meq gene of the isolate revealed >99% nucleotide sequence identity to virulent and very virulent plus strains from a number of European countries, placing it in the same subclade of clade III as two virulent Italian strains and a very virulent plus Polish strain as well as virulent strains of geese and ducks. The data reported here indicate that a virulent strain of Marek's disease virus is circulating in Turkey and has not been stopped by the current national vaccination programme.

Circulation of Indigenous Bovine Respiratory Syncytial Virus Strains in Turkish Cattle: The First Isolation and Molecular Characterization.

Bovine respiratory disease (BRD) is a huge economic burden on the livestock industries of countries worldwide. Bovine respiratory syncytial virus (BRSV) is one of the most important pathogens that contributes to BRD. In this study, we report the identification and first isolation, with molecular characterization, of a new BRSV strain from lung specimens of three beef cows in Turkey that died from respiratory distress. After the screening of lung tissues for BRD-associated viruses using a multiscreen antigen-ELISA, a BRSV antigen was detected. This was then confirmed by real-time RT-PCR specific for BRSV. Following confirmation, virus isolation was conducted in MDBK cell cultures and clear CPE, including syncytia compatible with BRSV, were detected. RT-nested PCR, using F gene-specific primers, was performed on the cultured isolates, and the products were sequenced and deposited to Genbank with accession numbers MT179304, MT024766, and MT0244767. Phylogenetic analysis of these sequences indicated that the cattle were infected with BRSV from subgroup III and were closely related to previously identified American and Turkish strains, but contained some amino acid and nucleotide differences. This research paves the way for further studies on the molecular characteristics of natural BRSV isolates, including full genome analysis and disease pathogenesis, and also contributes to the development of robust national strategies against this virus.

A Study on the Identification of Five Arboviruses from Hematophagous Mosquitoes and Midges Captured in Some Parts of Northern Turkey.

BACKGROUND: Whether zoonotic or not, arboviral infections are continuing to be a major threat to human health as well as the livestock industry all around the world. This project presented the results of the identification study on five arboviruses, including West Nile virus (WNV), Bovine ephemeral fever virus, Akabane virus, Bluetongue virus, and Epizootic hemorrhagic disease virus, in mosquitos and midges from eight provinces of the Black Sea Region. METHODS: During 2011 and 2012, 3193 mosquitoes were captured around natural streams, rivers, lakes, and ponds using dry-baited miniature light-traps. Identification studies were concluded by employing molecular methods. RESULTS: According to the morphological identification, blood-sucking mosquitoes and biting-midges belonged to Aedes (44.69%), Anopheles (28.34%), Culex (22.14%) and Culicoides (4.83%) species. Overall, 146 pools were made up of captured mosquitos and midges. None of the five viruses were directly identified by mosquitoes. CONCLUSION: Mosquitoes and midges have got a crucial role in the transmission of arboviruses. The risk of occurrence for the investigated arboviruses will continue depending upon many factors including the presence of these viruses in Turkey and its neighboring countries, uncontrolled livestock movements, global warming and climate changes.

Ebola virus disease and the veterinary perspective.

Ebola virus disease (EVD) is a potentially fatal haemorrhagic disease of humans. The last and most serious outbreak of Ebola virus (EBOV) started in December 2013 in West Africa and also affected other continents. Animals such as fruit bats and non-human primates are potential sources of EBOV. This review highlights the clinical features of EVD in humans and animals and addresses the public health implications of EVD outbreaks from the veterinary perspective.

Molecular detection of pestiviruses in aborted foetuses from provinces in northern Turkey.

All pestiviruses are important veterinary pathogens causing economic losses in cattle, sheep and pigs. Besides the important economical losses, pestiviruses may compromise the normal immune response to other pathogens and increase the severity of other infections in sheep. In this study, aborted foetuses (cattle and sheep) in either coastal or inland Black Sea region of Turkey were surveyed for the presence of RNA from pestiviruses (bovine viral diarrhoea virus (BVDV), border disease virus (BDV)). The presence of BVDV RNA was found in 6 of 21 aborted calves (28.57%), although BDV RNA was detected in 14 of 21 aborted lambs (66.66%) by reverse transcriptase polymerase chain reaction. This study also investigates the distribution of viral RNA within the brain, liver and lung of aborted foetuses. The viral RNA positivity rates for the organs varied and were as follows: brain 40.47% and liver and lung 38.09%. The results revealed that pestiviruses are important abort pathogen in the provinces of northern Turkey.

The first investigation of west nile virus in horses using real time rt-PCR in middle black sea region in Turkey.

BACKGROUND: West Nile Virus (WNV) is a mosquito-borne disease that can cause fatal infection in mammals including humans, dogs, horses, birds and reptiles. Although West Nile Virus is an asymptomatic infection, especially it can cause neurologic disorders in humans and horses. The aim of this study was to the investigate virological presence of WNV in horses in the Black Sea Region of Turkey using real time RT-PCR (rRT-PCR). METHODS: Totally, 120 horse sera were collected equally from 4 provinces in Middle Black Sea Region of Turkey and investigated for WNV presence by Taqman based rRT-PCR. RESULTS: WNV nucleic acid was not detected in any horse serum sample. CONCLUSION: Although obtained result indicated no evidence of WNV-RNA in horses, Black Sea Region of Turkey is one of the suitable places for the WNV infection. For this reason, our research will continue for the determination of the viruses in vectors and susceptible animals such as horses, dogs, etc.

Maedi-visna virus infection in Karayaka and Amasya Herik breed sheep from provinces in northern Turkey.

Maedi-visna is an important virus infection of sheep having prolonged incubation period (slow disease) and reflecting two distinct forms clinically and pathologically. In this study, the presence of MVV was investigated serologically in 58 Amasya Herik sheep breed and 525 Karayaka sheep breed. Seropositivity rates in Amasya Herik sheep breed and Karayaka sheep breed were detected as 69.0% and 18.5%, respectively. MVV antibodies were found in 137 of 583 serum samples (23.5%). Positivity rates for the provinces varied and were as follows: Samsun 19.4%, Sinop 15.4%, Ordu 25.8%, Trabzon 26.7%, Rize 36.7%, Amasya 69.0% and Tokat 35.0%, however no antibody response was detected in all of the sheep in Giresun province.

Distribution of HSV-1 IgG antibodies by two methods comparing in Turkish atopic children.

Herpes Simplex Virus Type 1 (HSV-1) is a ubiquitous pathogen. Other than known diseases, HSV-1 may have an important role in the pathogenesis of atopy by causing immortality of th2 cells. From June 1st to July 31st 2006, seventy five blood samples were collected from atopic children referred to the allergy clinic of the hospital. The bloods samples were used to detect HSV-1 IgG antibodies by Enzyme-Linked Immunosorbent Assay and Virus Neutralization Test. HSV-1 IgG antibody seroprevalence in atopic children was found high, 62.6% by Enzyme-Linked Immunosorbent Assay and 57.3% by Virus Neutralization Test. Thus Virus Neutralization Test sensitivity was 92.15% and specificity was 100% regarding to the Enzyme-Linked Immunosorbent Assay technic. Although Enzyme-Linked Immunosorbent Assay was more sensitive than Virus Neutralization Test, there was no significant difference between two technics (p > 0.05) in detecting HSV-1 IgG antibodies in serum.