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Öğe An assessment of bovine herpes virus 4 as a causative agent n abortions and neonatal death(Aosis, 2020) Dagalp, Seval B.; Babaoglu, Ali R.; Dogan, Firat; Farzani, Touraj A.; Alkan, FerayNumerous viruses, including bovine viral diarrhoea virus (BVDV), bovine herpes virus 1 (BoHV-1) and bovine herpes virus 4 (BoHV-4), and other pathogens are the most common causes of reproductive disorders and are responsible for huge economic losses in livestock production. This study investigates the aetiological role of BoHV-4 in fertility problems such as abortions, stillbirth and birth with unviable calves. Retrospective samples from 38 animals, including 17 aborting cows, 17 aborted foetuses, three stillborn calves and one unviable newborn calf were analysed. The BoHV-4 genome was detected in 25 (65.7%) animals by polymerase chain reaction. In 14 of these infected animals, we detected co-infection with BVDV, while the co-presence of BoHV-1 was also detected in one animal. In addition to the high prevalence of BoHV-4 genome in materials related to fertility problems, isolation of Bo H V-4 from the brain of one stillborn calf indicated a causal link between BoHV-4 and fertility problems, such as abortion, stillbirths or birth with unviable calves.Öğe Assessment of replication of bovine herpesvirus type 4 in human glioblastoma and breast cancer cells as a potential oncolytic virus(Springer, 2021) Aligholipour Farzani, Touraj; Bilge Dagalp, Seval; Ozkul, Aykut; Gurdal, Hakan; Dogan, Firat; Alkan, FerayOncolytic viruses have been extensively used in cancer treatment due to their tropism, selective replication only in tumor cells, and possible synergic interaction with other therapeutics. Different researchers have demonstrated that bovine herpesvirus 4 (BoHV-4), a member of the gammaherpesviridae family, has oncolytic potential in some human-origin cancer cell lines like glioma through the selective replication strategy. Using four apoptosis detection methods, namely MTT, LDH, TUNEL, and Annexin V assays, we evaluated the apoptotic effect of BoHV-4 Movar33/63 reference strain along with a recombinant BoHV-4 expressing EGFP in U87 MG cells (human glioblastoma cell line), MDA MB-231 (human breast cancer cell line), and MCF10a (non-tumorigenic human mammary epithelial cell line). Our findings indicate that this virus can replicate and induce apoptosis in these cell lines and hinder in vitro proliferation in a dose-dependent manner. In conclusion, BoHV-4 has in vitro potential as a novel oncolytic virus in human cancer therapy. However, its replication potential in the MCF10a cells as a non-tumorigenic human mammary epithelial cell line is a concern in using this virus in cancer therapy, at least against human mammary tumors. Further studies must therefore be conducted to examine the specific apoptotic pathways induced by this virus to move on to further experiments.Öğe 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, FerayBovine 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.Öğe Determination of presence of equid alpha and gammaherpesvirus infections in foals with respiratory distress(2018) Dağalp, Seval Bilge; Babaoğlu, Ali Rıza; Ataseven, Veysel Soydal; Karapınar, Zeynep; Timurkan, Mehmet Özkan; Doğan, Fırat; Özkul, Aykut; Alkan, FerayBu çalışmanın amacı, özel bir pansiyon harasında (geçici konaklama yeri) bulunan taylarda bir solunum sistemi hastalığı salgınında equid herpesvirusların varlığını araştırmak ve equid alpha ve gammaherpesvirusların olası etkileşimlerini ortaya koymaktır. Bu amaçla, solunum sistemi hastalığı bulgulu 28 taydan nazal swap (n=21), lökosit örnekleri (n=28) ve ölen bir taydan alınan doku örnekleri, equid herpesvirus-1, -4, -2 ve -5 yönünden glikoprotein B (gB) genini tespit etmeyi hedefleyen multipleks nested PCR ile test edildi. Örneklenen 29 hayvanın %3.4'ü (1/29), %58.6'sı (17/29), %58.6'sı (17/29) ve %75.9'u (22/29) sırasıyla EHV-1, EHV-4, EHV-2 ve EHV-5 yönünden pozitif olarak bulundu. Özellikle söz konusu viruslar tarafından oluşturulan tekli enfeksiyonlardan çok, EHV-1,-2 ve -5 arasında çoklu enfeksiyonların varlığı (%75.9; 22/29) dikkat çekiciydi. Filogenetik analiz sonuçları yerel equid gammaherpesvirusların, EHV-4 suşlarının aksine genetik olarak büyük farklılık gösterdiğini ortaya koyduÖğe Development of a BoHV-4 viral vector expressing tgD of BoHV-1 and evaluation of its immunogenicity in mouse model(Springer, 2021) Bilge-Dagalp, Seval; Farzani, Touraj Aligholipour; Dogan, Firat; Yoldar, Zeynep Akkutay; Ozkul, Aykut; Alkan, Feray; Donofrio, GaetanoIn recent years, Bovine herpesvirus 4 (BoHV-4) has emerged as an attractive gene delivery viral vector, mainly for vaccination purposes in the veterinary field. In the present study, a new infectious clone of the BoHV-4 genome carrying a bacterial artificial chromosome vector (BoHV-4-BAC) was developed by homologous recombination in mammalian cell culture and bacterial systems, and exploited to express a truncated form of glycoprotein D (tgD) of Bovine herpesvirus 1 (BoHV-1) (BoHV-4-tgD increment TK) as a vaccine candidate. This construct's immunogenicity was compared to a DNA vector expressing the same antigen (pC-tgD) in a BALB/c mouse model. After the mice were immunized, total and specific antibody responses, cytokine responses, total splenocyte cells proliferation/cytotoxicity, and virus neutralization assays were conducted to analyze the immune response elicited by both constructs. Mice from both vaccine groups developed significant humoral and cellular immune responses after a booster dose regime was conducted on day 28 post-injection. In almost all immunological assays, BoHV-4-tgD Delta TK induced as high an immune response as pC-tgD. In both vaccine constructs, neutralizing antibodies were a significant determining factor in protection against BoHV-1, even after the first injection. We conclude that a BoHV-4-based viral vector offers an effective immunization strategy as an alternative to DNA-based immunization platforms, at least to combat BoHV-1.Öğe 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, FerayBovine 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.Öğe The investigation of the presence of bovine herpesvirus type 4 (BoHV-4) in cows with metritis in a dairy herd(2010) Bilge Dağalp, Seval; Güngör, Elvin; Demir, Ayşe Başak; Muz, Dilek Pınar; Yılmaz, Volkan; Oğuzoğlu, Tuba Çiğdem; Ataseven, Veysel Soydal; Alkan, FerayTo investigate the occurrence of bovine herpesvirus type 4 (BoHV-4) infection in a dairy herd including cows with metritis, various methods were used for the diagnosis BoHV-4 in vaginal discharge samples. Blood serum samples were obtained from 368 cows for serodiagnosis of BoHV-4 infection. Of 368 serum samples, 256 (69.6%) were found positive for BoHV- 4 specific antibodies. For the virological study, vaginal discharge samples of 55 cows with metritis were used and tested for BoHV-4 using polymerase chain reaction (PCR), virus isolation and immunofluorescence techniques. Twenty-nine percent (16/55) of the vaginal discharge samples obtained from cows with metritis were found positive for BoHV-4 DNA by PCR. In this study, the presence of BoHV-4 was determined in cows with metritis in post-partum period in a dairy herd.Öğe 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, AykutThe 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.Öğe Molecular characterization of Bovine virus diarrhea viruses species 2 (BVDV-2) from cattle in Turkey(Springer, 2010) Oguzoglu, Tuba Cigdem; Muz, Dilek; Yilmaz, Volkan; Alkan, Feray; Akca, Yilmaz; Burgu, IbrahimFive BVDV species 2 (BVDV-2) isolates were detected from cattle in Turkey. Phylogenetic analysis was performed based on the 5' untranslated regions (UTRs) and E2 coding gene regions, respectively. The isolates were closely related to BVDV-2a strains from North America and Canada used as references. This is the first report of the detection of BVDV-2 in naturally infected Turkish cattle. It is important to consider BVDV-2 for planning future BVDV control and vaccination programs in Turkey.Öğe Porcine circovirus type 2 infection in Turkey(2011) Karaoğlu, Taner; Oğuzoğlu, Tuba Çiğdem; Ataseven, Veysel Soydal; Tan, Mehmet Tolga; Alkan, FerayPorcine circovirus is the smallest known DNA virus and is identified and characterized by two types: PCV type 1 (PCV 1) and PCV type 2 (PCV 2). Porcine circovirus type 1 (PCV 1) was first identified in 1974 and was recognized as a nondisease- causing agent that frequently occured in laboratory tissue cultures. As for PCV 2, it is defined as an antigenically and genomically different PCV which was commonly seen in swine populations and led to clinic disorders at the end of the 1990s. PCV2 infection in pigs can cause a wide variety of clinical signs and syndromes. This study aims to analyze the presence of PCV 2 related to different clinic cases and syndromes found on some pig farms. To this end, 86 nasal swab samples from two different pig farms and 12 lung tissue pieces from wild boars were collected to detect whether PCV 2 was present. In the study, 98 samples were used in total. The one step PCR technique was applied to the samples obtained. The samples were analyzed in terms of PCV presence; 38 of the samples were identified as PCV positive. Thirty-one of the 38 samples which were identified as PCV positive were identified as PCV 2 in the discriminant diagnosis, while 7 were evaluated as PCV 1.Öğe A teat papillomatosis case in a Damascus goat (Shami goat) in Hatay province, Turkey: a new putative papillomavirus?(Springer Wien, 2018) Dogan, Firat; Dorttas, Selvi Deniz; Bilge Dagalp, Seval; Ataseven, Veysel Soydal; Alkan, FerayPapillomaviruses (PVs) are epitheliotropic viruses that cause benign proliferative lesions in the skin (warts or papillomas) and mucous membranes of their natural hosts. Recently, new PVs have been found in many animal species. The most common current approach for identifying novel PV types is based on PCR, using various consensus or degenerated primer (broad-range primers), designed on the basis of the multiple alignment of nucleotide or amino acid sequences of a large number of different human papillomaviruses (HPV). PVs have been classified according to the sequence similarity of one of their capsid proteins, L1, without taking into account other regions of the genome and without considering the phenotypic characteristics of the viral infection. In this study, we performed molecular detection and typing of a PV in a goat with teat papillomatosis. Firstly, PCR was performed using the FAP59/FAP64 and MY09/MY11 primer pairs for the L1 gene region. The PV DNA was found to be positive only with the FAP59/FAP64 primer pair. PV DNA was then tested with three primer sets in four different combinations (L2Bf/FAP64, L2Bf/L1Br, FAP59/FAP64, L1Bf/LCRBr) for the gene region encoding the L1, L2 and LCR proteins. The goat teat papilloma sample was amplified using FAP59/FAP64 primers and two primer pairs (L2Bf/FAP64 and L2Bf/L1Br). We obtained products matching approximately 604 bp of the L1 region of the virus. PV DNA was used for typing using sequence analysis/PCR with some type-specific primers for bovids, caprids and cervids. The results of the sequence analysis suggested one new putative PV type with sequence identity ranging from 46.45 to 80.09% to other known papillomaviruses, including Capra hircus papillomavirus (ChPV-2), bovine papillomavirus (BPV) 6, 7, 10, 11 and 12, Rangifer tarandus papillomavirus 3 (RtPV-3) and BPV-7Z (Alpine wild ruminant papillomavirus; Cervus elaphus papillomavirus). We therefore propose that this is the first identification of a new putative type, MG523274 (HTY-goat-TR2016), in a goat with teat papillomatosis. It is essential to identify PV types in different animal species and investigate their prevalence/distribution and clinical consequences in order to develop appropriate prophylactic and/or therapeutic procedures and to determine the interspecies transmission potential and evolution of PVs.
