Person: YEŞİLBAĞ, KADİR
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YEŞİLBAĞ
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KADİR
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Publication Molecular characterization and comparison of diagnostic methods for bovine respiratory viruses (BPIV-3, BRSV, BVBV, and BoHV-1) in field samples in northwestern Turkey(Springer, 2021-01-06) Toker, Eda Baldan; Yeşilbağ, Kadir; TOKER, EDA BALDAN; YEŞİLBAĞ, KADİR; Bursa Uludağ Üniversitesi/Veteriner Fakültesi/Viroloji Anabilim Dalı; 0000-0003-2468-3945; ABE-9974-2020; ABE-7662-2020The aim of this study was to evaluate the compatibility among virus isolation (VI), ELISA, and PCR for diagnosis of the major viral agents (BPIV-3, BRSV, BVDV, and BoHV-1) responsible for BRD in the field samples. For that purpose, a total of 193 samples (133 nasal swabs and 60 lung tissue samples) from cattle with respiratory signs in northwestern Turkey were examined. For VI, all the samples were inoculated at least 3 blind passages onto MDBK cell culture. In addition, the samples were tested by hemadsorption assay and RT-PCR for BPIV-3; nested RT-PCR for BRSV; immunoperoxidase monolayer assay, antigen-ELISA, and RT-PCR for BVDV; and antigen-ELISA and PCR for BoHV-1. The detected 1 (0.52%) BPIV-3 isolate was found to be in the genotype BPIV-3c. No BRSV isolate could be obtained, while 5 (2.59%) samples were evaluated positive in nested-RT PCR. The presence of BVDV antigen in 10 (5.18%) samples and the BVDV genome in 5 (2.59%) samples were detected, while non-cytopathogenic BVDV isolates were obtained only in 2 (1.04%) samples. The detected BVDV strains fell into the genetic clusters of BVDV-1a, -1f, and -1l. For detection of BoHV-1, although viral isolation and Ag-ELISA results were negative, presence of BoHV-1.1 genome was detected in 2 (1.04%) samples. By the results of VI, ELISA, and PCRs, 10.88% (21/193) of samples were found positive for the evaluated viruses. Depending on the obtained data, combined uses of the diagnostic methods were evaluated to be more reliable for routine diagnosis of bovine respiratory viruses.Publication Ivermectin also inhibits the replication of bovine respiratory viruses (BRSV, BPIV-3, BoHV-1, BCoV and BVDV) in vitro(Elsevier, 2021-03-05) Yeşilbağ, Kadir; Toker, Eda Baldan; Ateş, Özer; YEŞİLBAĞ, KADİR; TOKER, EDA BALDAN; Ateş, Özer; Bursa Uludağ Üniversitesi/Veteriner Fakültesi/Viroloji Anabilim Dalı.; 0000-0003-2468-3945; 0000-0001-7676-9033; ABE-9974-2020; AAH-3493-2021; GJX-0664-2022Bovine respiratory disease (BRD) complex is an important viral infection that causes huge economic losses in cattle herds worldwide. However, there is no directly effective antiviral drug application against respiratory viral pathogens; generally, the metaphylactic antibacterial drug applications are used for BRD. Ivermectin (IVM) is currently used as a broad-spectrum anti-parasitic agent both for veterinary and human medicine on some occasions. Moreover, since it is identified as an inhibitor for importin ?/?-mediated nuclear localization signal (NLS), IVM is also reported to have antiviral potential against several RNA and DNA viruses. Since therapeutic use of IVM in COVID-19 cases has recently been postulated, the potential antiviral activity of IVM against bovine respiratory viruses including BRSV, BPIV-3, BoHV-1, BCoV and BVDV are evaluated in this study. For these purposes, virus titration assay was used to evaluate titers in viral harvest from infected cells treated with noncytotoxic IVM concentrations (1, 2.5 and 5 ?M) and compared to titers from non-treated infected cells. This study indicated that IVM inhibits the replication of BCoV, BVDV, BRSV, BPIV-3 and BoHV-1 in a dose-dependent manner in vitro as well as number of extracellular infectious virions. In addition, it was demonstrated that IVM has no clear effect on the attachment and penetration steps of the replication of the studied viruses. Finally, this study shows for the first time that IVM can inhibit infection of BRD-related viral agents namely BCoV, BPIV-3, BVDV, BRSV and BoHV-1 at the concentrations of 2.5 and 5 ?M. Consequently, IVM, which is licensed for antiparasitic indications, also deserves to be evaluated as a broad-spectrum antiviral in BRD cases caused by viral pathogens.Publication Persistent BVD virus infections in offspring from imported heifers(Springer, 2019-02-01) Alpay, Gizem; Toker, Eda Baldan; Yeşilbağ, Kadir; Alpay, Gizem; TOKER, EDA BALDAN; YEŞİLBAĞ, KADİR; Bursa Uludağ Üniversitesi/Veteriner Fakültesi/Viroloji Anabilim Dalı.; 0000-0002-3411-081X; 0000-0003-2468-3945; ABE-9974-2020; AAH-3917-2021; ABE-7662-2020The aim of this study was to investigate the possible risk of bovine viral diarrhea virus transport from imported live animals. For this purpose, two different groups of animals were sampled in this study. Group 1 consisted of pregnant heifers; group 2 consisted of male beef cattle imported during 2011-2012 and 2015, respectively. Blood samples were tested for pestivirus antigen using a commercial BVDV antigen ELISA. All the pregnant heifers were negative, but 9 out of 412 offspring and 5 of the 332 male cattle were BVDV antigen positive. Virus isolation and also investigation by RT-PCR were carried out by using 14 ELISA-positive samples. At the end of three blind passages, eight non-cytopathogenic isolates were obtained by indirect immunoperoxidase monolayer assay, which were also RT-PCR positive using panpesti-virus primers. After discriminative RT-PCR, all the isolates that were identified as BVDV-1 and 5UTR-based analysis demonstrated the existence of BVDV-1b (n=4), BVDV-1f (n=2), BVDV-1l (n=1), and BVDV-1r (n=1) subgenotypes. There was no BVDV subgroup that is newly introduced into the country. However, detection of persistent infection in calves born from imported animals demonstrates the risk of BVDV virus introduction by imported animals into the receiving country. Viral strains from persistently infected animals were characterized as BVDV-1b, which is predominant subgroup in the country where animals are imported. These results highlight a possible problem for the areas where a BVDV control program is currently ongoing. Additionally, sequences obtained in this study also showed that there are two distinct branches identified in BVDV-1l.Publication In vitro antiviral activity of thymbra spicata L. extract on bovine respiratory viruses (BCoV, BPIV-3, BRSV, BVDH and BoHV-1)(Oxford Univ Press, 2021-12-21) Toker, Eda Baldan; Yeşilbağ, Kadir; TOKER, EDA BALDAN; YEŞİLBAĞ, KADİR; Bursa Uludağ Üniversitesi/Veteriner Fakültesi/Viroloji Anabilim Dalı; 0000-0003-2468-3945; ABE-9974-2020; GJX-0664-2022Aims Viral pathogens are the primary agents in bovine respiratory disease cases, and there is no direct effective antiviral drug application. Thymbra is a genus of oregano commonly found in Turkey. The primary component (34.9%) of the extract obtained from Thymbra spicata L. is the carvacrol which is used in traditional medicine. This study evaluates the potential antiviral activity and inactivation efficiency of T. spicata L. extract against bovine respiratory viruses, including BCoV, BPIV-3, BRSV, BVDV and BoHV-1. Methods and Results To evaluate its effect on viral replication, viral titres were taken from infected cells treated with non-cytotoxic T. spicata L. extract concentrations (0.75% and 1.5%, 1.32 and 2.64 mu g/ml of carvacrol as active ingredient, respectively) and compared to non-treated infected cells. The viruses were treated directly with 1.5% T. spicata L. extract, and the viral titres were evaluated at certain time points to determine the efficiency of direct inactivation. The number of infectious virions for BCoV, BPIV-3, BRSV, BVDV and BoHV-1 treated with 1.5% T. spicata L. extract were decreased by 99.44%, 100.0%, 94.38%, 99.97% and 99.87%, respectively.T. spicata L. extract strongly inhibits the replication of mentioned viruses in a dose-dependent manner in vitro. In addition, T. spicata L. extract shared direct inactivation efficiency on the mentioned viruses in a time-dependent manner. Conclusion This study shows the antiviral efficiency of T. spicata L. on BRD-related viral agents for the first time. The oregano species T. spicata and its main component, carvacrol, may have a potential for antiviral activity in the alternative treatment of respiratory viral diseases in cattle. Significance and Impact of the Study Given the similarity of replication strategies, obtained data suggest the possible efficiency of T. spicata L. on human respiratory viruses.Publication Failure in dry period vaccination strategy for bovine viral diarrhea virus(Elsevier, 2020-07-10) Toker, Eda Baldan; Aytoğu, Gizem; Kadiroğlu, Berfin; Ateş, Özer; Yeşilbağ, Kadir; TOKER, EDA BALDAN; AYTOĞU, GİZEM; KADİROĞLU, BERFİN; Ateş, Özer; YEŞİLBAĞ, KADİR; Bursa Uludağ Üniversitesi/Veteriner Fakültesi/Viroloji Anabilim Dalı.; 0000-0003-2468-3945; 0000-0001-7676-9033; AAH-3493-2021; AAH-3875-2021; ABE-7662-2020; ABE-9974-2020; ELC-2891-2022Bovine viral diarrhea is a common disease of cattle and has significant impact on animal welfare worldwide. There are fundamental approaches i.e. elimination of persistently infected animals, vaccination and biosecurity measures for effective control and eradication of BVD virus (BVDV). By this study, the presence of persistent infection with divergent BVDV subgenotype in the calves in a dairy herd having regular vaccination program was investigated. In the herd, vaccinated with a killed whole virion trivalent vaccine (composed of BVDV-1a) during the dry period of the cows, abortion cases were existed in the late autumn 2019. During herd screening by BVDV antigen-ELISA, 2 out of 300 dams were detected positive. Following, by ear notch-based BVDV antigen-ELISA, 30 calves were detected positive. Confirmation of persistent BVDV infection was performed 3 weeks later by testing with antigen-ELISA, where 8 of 9 selected newborn calves were positive for the second time. The entire antigen-ELISA positive samples were subjected to virus isolation on MDBK cell culture and identified as non-cytopathogenic pestiviruses by indirect immunoperoxidase assay. Presence of pestivirus RNA was detected in the 8 isolates by panpestivirus RT-PCR. Analysis of the 5'UTR regions revealed that BVDV-1 r circulate in the herd. Results of this study lead to questioning the efficiency of dry period vaccination strategy against BVDV. But otherwise, vaccination with BVDV-1a can be inefficient for complete protection against BVDV-1 r. Therefore, serological relationship between mentioned subgenotypes or protection by current vaccines against latest field isolates needs to be investigated before development of new BVDV vaccine candidates.Publication Neutralizing antibody titers against field strains of bovine viral diarrhea virus after vaccination with three commercial vaccines(Tübitak Bilimsel ve Teknolojik Araştırma Kurumu, 2019-01-01) Alpay, Gizem; Yeşilbağ, Kadir; Alpay, Gizem; YEŞİLBAĞ, KADİR; Uludağ Üniversitesi/Veteriner Fakültesi/Viroloji Anabilim Dalı; 0000-0002-3411-081X; 0000-0003-1793-6879; AAH-3917-2021; ABE-7662-2020Modified killed vaccines against bovine viral diarrhea virus (BVDV) are used worldwide. In the present study, the cross-neutralization antibody responses against field strains from seven subgenotypes of BVDV-1 and one subgenotype of BVDV-2 were evaluated using sera obtained by three inactivated commercial vaccines. One vaccine contained both BVDV-1 and BVDV-2 strains, while the others contained only BVDV-1a. Three vaccine groups, each containing five calves, were vaccinated two times with 30-day intervals. The antibody titers were evaluated by virus neutralization assay. The monovalent vaccine induced the highest antibody titers. Significant levels of neutralizing antibody titers were maintained up to the last sampling time. Although one vaccine contained the BVDV-2 strain, the lowest antibody titers were detected against field strains from BVDV-2, BVDV-1b, and BVDV-1r. This study indicated that cross-protective immune responses with the most common international BVDV vaccine strains need to be evaluated with challenge experiments against field strains for efficient protection.Publication Characterization of bovine rotavirus isolates from diarrheic calves in Turkiye(Springer, 2023-01-23) Ateş, Özer; Yeşilbağ, Kadir; Ateş, Özer; YEŞİLBAĞ, KADİR; Bursa Uludağ Üniversitesi/Veteriner Fakültesi/Viroloji Anabilim Dalı; 0000-0001-7676-9033; AAH-3493-2021; GJX-0664-2022BackgroundNeonatal calf diarrhea, which is the most common cause in calf deaths, leads to significant economic losses in dairy farming around the world. Diarrhea develops due to infectious and non-infectious reasons. Group A Rotaviruses (RVA) are the leading and predisposing factor for acute neonatal gastroenteritis.Methods and resultsIn this study, 20 diarrheic fecal samples were collected from one farm in Balikesir province of Turkey. During virus isolation, a total of 2 stool samples were detected to produce cytopathogenic effects in MA-104 cell line. The two samples (RV-36, RV-38) were tested positive with antigen ELISA kits detecting RVA antigens. In order to detect the presence of rotavirus viral nucleic acid in cell supernatants, VP6 gene region-specific RT-PCR test was performed and the samples RV-36 and RV-38 were positive for RVA viral nucleic acid. By RT-PCR using genotype specific primers, both the isolates RV-36 and RV-38 formed amplicons compatible with G10 and P[11] genotypes of RVA. RVA nucleic acids segments were also visualized by poliacrilamide gel electrophoresis (PAGE) method. The phylogenetic tree constructed according to the VP6 gene region showed that these isolates were in the Rotavirus A group and in the I2 cluster same as other bovine and some human RVA isolates.ConclusionSuccesful isolation of RVA G10P[11] was echieved in the cattle farm. As rotaviruses play the most important role in the etiology of diarrhea in newborn calves respected genotype G10P[11] should be considered in selection of the vaccines applied to the dams. Those isolates can be further evaluated as vaccine candidate.Publication Pathogenicity assessment of a bovine viral diarrhea virus type 1l (bvdv-1l) strain in experimentally infected calves(Elsevier, 2023-12-27) Kadiroğlu, Berfin; Ateş, Özer; Aytoğu, Gizem; AYTOĞU, GİZEM; Yeşilbağ, Kadir; YEŞİLBAĞ, KADİR; Toker, Eda Baldan; Yaşar, Mevlüt; TOKER, EDA BALDAN; Bursa Uludağ Üniversitesi/Veteriner Fakültesi.; 0000-0003-2468-3945; AAH-3493-2021Bovine viral diarrhea is a widespread and economically important viral disease for livestock which can cause clinically diverse manifestations. The number of established BVDV subgenotypes has increased, not only the serological relationships of recently described subgenotypes but virulence and pathogenic characteristics have not yet been mostly elaborated. The dominant BVDV subgenotype in Turkiye was elaborated to be BVDV-1l, that involves more than half of field strains and there is no scientific data to identify the pathogenicity of this strain so far. This study investigated the pathogenicity of a selected field strain (TR-72) from subgenotype BVDV-1l. Experimental infection was implemented by intranasal inoculation of the strain TR-72 (10 x105.5) to four young calves which were previously not vaccinated and were free both for BVDV antibodies and antigens. Clinical changes as well as blood parameters, body temperature, and viremia were monitored for 14 days. Only mild clinical signs associated with respiratory signs of BVDV infection were observed. Detected clinical signs included nasal discharge, conjunctivitis, cough, fatigue, high rectal temperature reaching 40.7 celcius, and white blood cell counts depression started from the 2nd day and 40.4% decreased between the 12th and 14th days post-infection (poi). The presence of viremia was investigated by virus isolation, RT-PCR, and real-time RT-PCR from blood samples. The efficiency of experimental infection was established not only by observed clinical signs but also by virus isolation from blood leukocytes between the 5th and 8th days poi., virus detection was obtained by realtime PCR between the 3rd - 13th days poi. Besides, the recorded mild clinical signs, high fever, long duration of viremia , and high decrease in blood parameters obtained in this study, it was shown that the noncytopathogenic BVDV-1l strain TR-72 has a moderate virulence in naive cattle.Publication First isolation and molecular characterization of pseudorabies virus detected in Turkey(Springer, 2022-01-15) AYTOĞU, GİZEM; YAVAŞ, ÖZKAN; KADİROĞLU, BERFİN; TOKER, EDA BALDAN; YEŞİLBAĞ, KADİR; Ateş, Özer; ÖZYİĞİT, MUSA ÖZGÜR; Bursa Uludağ Üniversitesi/Veteriner Fakültesi/Viroloji Anabilim Dalı.; Bursa Uludağ Üniversitesi/Veteriner Fakültesi/Patoloji Anabilim Dalı.; 0000-0003-2468-3945; 0000-0001-7676-9033; KHD-4075-2024; AAH-3493-2021; ABE-9974-2020Background Pigs are the main host species for the pseudorabies virus. It causes fatal encephalitis in many species, including humans. This article aims to report the first clinical case of pseudorabies as well as isolation and molecular characterization of the virus from a hunting dog in Bursa province, Turkey. Methods and results The dog shows clinical signs including pruritus and neurological signs such as stumbling and inability to stand up compatible with pseudorabies. The virus isolates were obtained from the supernatant of fresh tissue samples from the cerebellum, cornu ammonis, spleen, salivary gland, conjunctival swab, serum, and PBMC samples. The glycoprotein C region is targeted for viral DNA amplification. Pseudorabies virus genome detected both in fresh tissues and supernatants of third passage on Vero cells. The number of PCR positive samples was dramatically increased after cell culture inoculations. Genome sequencing of strain Bursa-10303, which was isolated from a non-endemic area, identified it to belong to clade A. Conclusions This study confirms the possible presence of pseudorabies infection in the wildlife reservoirs in Turkey. Future studies may clarify the importance of the infection in Turkey region, where there is no prevalent pig production.Publication Effect of size on biaxial flexural strength for cement-based materials by using a triangular plate method(Elsevier, 2022-01-05) TOKER, EDA BALDAN; YEŞİLBAĞ, KADİR; Yeşilbağ, Kadir; Ateş, Özer; 0000-0003-2468-3945; 0000-0001-7676-9033; ABE-9974-2020; AAH-3493-2021Capripoxvirus diseases are listed as reportable diseases by World Organization for Animal Health (OIE). Lumpy skin disease virus (LSDV) and sheeppox virus (SPPV), which can only be distinguished by molecular analysis, cause moderately, severe, or sometimes fatal infections in cattle and sheep. Even though vaccines are the most effective way to control the infection, their effectiveness may decrease in some cases. Therefore, it is significant to explore antiviral drugs against these diseases along with the vaccine. This study aimed to investigate the antiviral efficiency of ivermectin (IVM) at different stages of in vitro replication of LSDV and SPPV. For this purpose, viral titers (TCID50/mL) of the viruses not treated with IVM (0.0 mu M) and treated with non-cytotoxic concentrations of IVM (1.0 and 2.5 mu M) were compared during a nine-day (216 h) post-infection period by viral titration assay. At 2.5 mu M concentrations of IVM, the mean viral titer was significantly (P < 0.05) reduced by approximately three logs for the replication stage of LSDV and SPPV. To evaluate the antiviral activity of IVM against LSDV and SPPV by treatment at the virus attachment and penetration stages, the titers of the virus either untreated or treated with 2,5 mu M IVM were compared by virus titration assay. The number of infectious virions for LSDV and SPPV were decreased by 99.82% and 99.87% at the viral replication stage, 68.38% and 25.01% at the attachment stage, and 57.83% and 0.0% at the penetration stage, respectively. It was determined that ivermectin is statistically more effective on LSDV than SPPV at the virus attachment and penetration stages (P < 0.05). This study found that the drug IVM can inhibit capripoxviruses, including LSDV and SPPV at various stages of the propagation. Moreover, this research predicted the in vitro antiviral ability of IVM against capripoxvirus infections for the first time.