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    Akabane virus infection in Eastern Mediterranean Region in Turkey: Culicoides (Diptera: Ceratopogonidae) as a possible vector
    (Springer, 2021) Dagalp, Seval Bilge; Dik, Bilal; Dogan, Firat; Farzani, Touraj Aligholipour; Ataseven, Veysel Soydal; Acar, Gulizar; Sahinkesen, Ilker
    Akabane virus (AKAV), which causes Akabane disease, is an arthropod-borne virus (arbovirus) transmitted by Culicoides biting midges and mosquitoes. AKAV is an important pathogen that causes abortion and congenital anomalies in ruminants. In this study, we determined the prevalence of AKAV infection and identified possible viral vectors in Turkey's Eastern Mediterranean region. The presence and prevalence of AKAV infection were assessed using serological and virological methods. Serologically, the prevalence of AKAV antibodies in cattle, sheep and goats were 44.74% (400/894), 22.90% (60/262) and 14.52% (63/434), respectively, while the total prevalence was 32.89% (523/1590). AKAV-specific nucleic acid amplicons were obtained by real-time RT-PCR from 1.13% (9/799) and 1.74% (5/288) of the cattle and sheep tested, respectively. No goats were positive for AKAV RNA. Overall, AKAV-specific nucleic acid amplicons were detected in 0.87% (14/1604) of the sampled ruminants. In addition, specimens of the assumed vector, Culicoides, were caught using light traps and identified. Ten Culicoides species were detected in the area, of which Culicoides schultzei complex was the dominant species although 32 specimens could not be identified at the species level. These were defined as Culicoides spp. AKAV nucleic acid was detected in C. schultzei, Culicoides longipennis and Culicoides circumscriptus. Phylogenetic analysis indicated two different AKAV genogroups (genogroups Ib and genogroups II) while potential AKAV vectors in this region are C. schultzei complex, C. longipennis and C. circumscriptus.
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    Detection of genotype 1 bovine leukemia virus from a C.schultzei pool: Do Culicoides spp. have a role on the transmission of bovine leukemia virus?
    (Elsevier, 2020) Dogan, Firat; Dagalp, Seval Bilge; Dik, Bilal; Farzani, Touraj Aligholipour; Alkan, Feray
    Bovine leukemia virus (BLV) is known as the etiological agent of Enzootic bovine leukosis (EBL), which is the most common neoplastic disease of cattle. While the major route of virus transmission is believed to be iatrogenic, BLV proviral DNA has been identified in biological materials, including nasal secretions, saliva, milk, colostrum, and semen, and in several insect species, including horses flies. However, insects' role in the natural transmission of BLV has not been clearly demonstrated. This study assessed the possible role of midges - Culicoides spp. - in BLV transmission. BLVs were genetically characterized and BLV infection seroprevelance was determined in 224 cattle sampled from 27 different small family herds in five different districts in Hatay province, southern Turkey. Out of the 25 Culicoides spp. pools, one (4.0%; 1/25) was a C.schultzei pool while 2.67% (6/224) of the sampled cattle were positive for BLV nucleic acid. The seroprevalance rates for the sampled herds and all sampled cattle were 7.40% (2/27) and 1.33% (3/224), respectively. According to the phylogenetic analysis, the sequences of the BLVs from the cattle (n = 6) and the one BLV-positive C.schultzei pool clustered on genotype 1 (G1) BLVs. Although these results do not reveal the exact role of Culicoides spp. or other midges flies in BLV transmission, the simultaneous presence of same substitions in BLVs from both cattle and a C.schultzei pool is noteworthy. Further studies on the env gene and other BLV gene regions detected from cattle and C.schultzei pools are ongoing to understand the possible epidemiological relationship between cattle and flies.
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    Equine and bovine papillomaviruses from Turkish brood horses: a molecular identification and immunohistochemical study
    (Univ Zagreb Vet Faculty, 2019) Kanat, Ozgur; Ataseven, Veysel S.; Babaeski, Serdar; Derelli, Fatih; Kumas, Cihan; Dogan, Firat; Dagalp, Seval Bilge
    Papillomaviruses have an oncogenic nature, developing benign or malignant tumours in association with the proliferation of cutaneous or mucosal epithelia in humans and animals. The objective of this study was to investigate, the genoprevalence of bovine papillomavirus (BPV) types 1 and -2. and equine papillomavirus (EcPV) types 1. 2 and 3 in Turkish brood horses, with or without genital lesions and skin tumours. and confirmation of the equine sarcoids by several immunohistochemical markers. A total of 42 genital swabs and 6 skin tumours were collected from Thoroughbred stallions/mares in Turkey. Overall, both EcPV and BPV specific DNA amplicons sampled were detected in genital swabs from 38.1% of the brood horses tested by PCR. The prevalence of BPV-1, BPV-2 and EcPV-2 in the Turkish brood horses was 14.3%, 2.4% and 21.4%, respectively. Furthermore, there was no significant difference between the horses that appeared to be healthy (38.7%) and symptomatic horses (36.4%) (P>0.05). Out of the six samples obtained from the six horses that had skin tumours, BPV-1 was detected in five tissue samples and four blood samples, and only one skin tumour was found to be infected with EcPV-2. This study indicates that a high prevalence of EcPV-2 and BPV-1 was found in apparently healthy horses as well as in symptomatic cases. Additionally, the horses infected subclinically with papillomaviruses may play a significant role in the epizootiology of papillomavirus infections and lead to an increase in reproductive problems in brood horse populations.
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    Genetic variability of bovine herpesvirus type 4 (BoHV-4) field strains from Turkish cattle herds
    (Ist Zooprofilattico Sperimentale Abruzzo & Molise G Caporale-Izs A&M, 2021) Dagalp, Seval Bilge; Dogan, Firat; Babaoglu, Ali Riza; Farzani, Touraj Aligholipour; Alkan, Feray
    Bovine herpesvirus type 4 (BoHV-4) is a common virus in the world that is detected in clinically ill or in apparently healthy cattle.This study provides a molecular characterization of BoHV-4 strains from 24 cattle some showing respiratory and/or reproductive problems and some without any apparent clinical sign. This study also reported the growth properties of five BoHV-4 field isolates. The 24 sampled cattle came from 13 different herds in 10 provinces collected between 2007 and 2018. Phylogeneticanalysis using partially amplified nucleotide sequences of ORF8 genes coding glycoprotein B (n = 24) and ORF3 genes coding thymidine kinase (n = 9), demonstrated genetic variability among the BoHV-4 strains analysed. The partial gB gene sequences clustered in three different genotypes (genotype 1,11and111) were located within the genotype I cluster, such as Movar strain. The analysis of the five BoHV-4 strains isolated from vaginal swabs (n = 2), nasal swab (n = 1), and brain samples (n = 2) revealed no significant differences in their growth properties in MDBK cell culture.
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    Molecular and antigenic characterization of bovine herpesvirus type 1 (BoHV-1) strains from cattle with diverse clinical cases in Turkey
    (Springer, 2020) Dagalp, Seval Bilge; Farzani, Touraj Aligholipour; Dogan, Firat; Alkan, Feray; Ozkul, Aykut
    The present study reports the molecular and antigenic characterization of 13 bovine herpesvirus type 1 (BoHV-1) field viruses obtained from cattle with different clinical cases in Turkey between 1992 and 2017. We selected glycoprotein C (gC) of BoHV-1 as a target to detect and/or verify presence of the virus in suspect materials followed by virus isolation (VI) in MDBK cells. In seven out of 13 BoHV-1 positive samples, cytophatic effects (CPEs) were observed in MDBK cell cultures, although only four virus samples reached a sufficient titer to use in phylogenetic assay, restriction endonuclease analysis (REA), and virus neutralization test (VNT). According to the results of sequence analysis of the 13 BoHV-1 positive samples, nine BoHV-1 field viruses were determined as BoHV-1.1 and four as BoHV-1.2. Using REA, we demonstrated that two of our isolated viruses could be categorized as BoHV-1.1 while the other two isolates were BoHV-1.2 subtypes. Differences between the BoHV-1.1 and BoHV-1.2 isolates were also detected in the VNT results by assaying 125 suspected serum samples after testing with isolated (KY748023, KY748022, KY748020, and KY748021) and reference viruses (BoHV-1 Cooper and BoHV-5 Texas 89). These results are indicating the need to correctly identify BoHV-1 field isolates to better understand the epidemiology and pathogenesis of infection. In addition, it would be useful to identify the subtypes circulating in the specific geographical area while determining vaccination preferences.