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    In vitro antiviral activity of thymbra spicata L. extract on bovine respiratory viruses (BCoV, BPIV-3, BRSV, BVDH and BoHV-1)

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    Aims 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

    Molecular characterization and comparison of diagnostic methods for bovine respiratory viruses (BPIV-3, BRSV, BVBV, and BoHV-1) in field samples in northwestern Turkey

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    The 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

    Recent strains of influenza D virus create a new genetic cluster for European strains

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    Bovine respiratory diseases (BRD) are one of the significant health problems for cattle breeding industry. Influenza D virus (IDV) alone or in combination with other respiratory pathogens plays a role in BRD. According to the IDV-HEF gene region, phylogenetic analyzes revealed five lineages: D/OK, D/660, D/Yama2016, D/ Yama2019, and D/CA2019, so far. In this study, despite no success in virus isolation, the presence of IDV was investigated by RT-PCR (partial HEF gene region) in 219 nasal swab samples collected from cattle with BRD between 2012 and 2021. The presence of IDV was demonstrated in two samples, and genome characterization data of the IDV sequences both in the partial and complete HEF gene regions showed that one of the obtained sequences (D/bovine/Turkey-Bursa/ET-138/2021) was in the lineage D/Yama2019 while the other (D/bovine/ Turkey-Bursa/ET-130/2013) created a new lineage tentatively called D/Bursa2013 as including few partial IDV sequences reported in Europe. Two nucleotide substitutions (nt252A-*G, nt299T-*C) were typically character-ized for the tentative lineage D/Bursa2013, one of which also leads to a unique amino acid change at position aa100 (V-*A). When the amino acid differences between the lineages were evaluated, amino acid substitution changes were detected in four regions [aa12 (Alanine-*Aspartic acid), aa19 (Glycine-*Arginine), aa22 (Proli-ne-*Serine), and aa110 (Aspargine-*Arginine)] of the D/Yama2019 lineage, unlike the other lineages. Consid-ering the most common D/OK lineage in Europe, many nucleotide substitutions were shown between D/OK and D/Bursa2013. Accordingly, aminoacid substitutions were observed in aa27 (Threonine-*Asparagine) and aa100 (Valine-*Alanine) in the D/bovine/Turkey-Bursa/ET-138/2021 sequence. Study results describe the circulation of D/Yama2019 and D/Bursa2013 (new lineage) in Turkey. Expansion of new strains seems possible due to the high mutation rate of influenza viruses. It is important to understand the development of IDV with compre-hensive characterization studies

    Persistent BVD virus infections in offspring from imported heifers

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    The 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

    Failure in dry period vaccination strategy for bovine viral diarrhea virus

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    Bovine 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

    Lumpy skin disease threat in Europe: Current situation, transmission dynamics and future prospects

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    Lumpy skin disease (LSD), caused by Lumpy Skin Disease Virus (LSDV, genus Capripoxvirus), is an emerging transboundary disease of cattle and water buffalo. Although rarely fatal, it results in severe economic losses due to decreased productivity, hide damage, infertility, and trade limitations. This review summarizes the etiology, susceptible hosts, modes of transmission, and advances in control, vaccination, and treatment strategies, with a particular focus on Europe, where spread into previously unaffected areas has recently been reported. Since its first identification in Zambia in 1929, LSD remained endemic in Africa for six decades before spreading into the Middle East, Asia, and Europe. Major outbreaks have subsequently occurred in Türkiye, the Balkans, Russia, Asian territories, and, most recently, in Italy, France, and Spain in 2025. Transmission is primarily vector-borne through blood-feeding arthropods, while animal movements and inadequate farm-level biosecurity further exacerbate the spread. Control measures such as vaccination with homologous live-attenuated vaccines, quarantine, and movement restrictions have proven effective in limiting outbreaks. LSD poses a significant threat to livestock health and trade globally. Coordinated surveillance, improved biosecurity, and vaccination remain the cornerstones of control. Continued research into antiviral and alternative therapeutic strategies to complement existing prevention measures is warranted. Killed vaccines may currently help to protect animals in pre-epidemic areas and slow the rate of epidemics

    Molecular diagnosis of bovine respiratory viruses in clinical and necropsy samples

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    Bu tez çalışmasında Bursa ve komşu illerde bulunan sığırlarda solunum sistemi enfeksiyonlarına neden olan önemli viral patojenlerden bovine respiratorik sinsityal virus (BRSV), bovine parainfluenza virus-3 (BPIV-3), bovine viral diarrhea virus (BVDV) ve bovine herpesvirus-1 (BoHV-1) etkenlerinin konvansiyonel virolojik yöntemler ve moleküler yöntemler aracılığıyla tespit edilmesi, genetik karakterizasyonunun yapılması ve saha izolatlarının elde edilmesi hedeflenmiştir. Bu amaçla solunum sistemi bulguları gösteren sığırlarda 133 nazal svab ve 60 akciğer doku örneği olmak üzere toplam 193 örnek değerlendirildi. Tüm örnekler MDBK hücre hattında virus izolasyonuna tâbi tutuldu. Ayrıca örneklere BRSV varlığını tespit etmek için nested RT-PCR; BPIV-3 için hemadsorbsiyon testi ve RT-PCR; BVDV için immunoperoksidaz testi, antijen-ELISA ve RT-PCR; BoHV-1 için antijen-ELISA ve PCR yöntemleri uygulandı. Virus izolasyonu çalışmalarında 1 BPIV-3 (ET-81) ve 2 non-sitopatojen BVDV saha izolatı (DO-48 ve ET-94) elde edildi. Antijen-ELISA testleri sonucunda 10 örnek BVDV antijen pozitif olarak saptanırken; BoHV-1 antijeni tespit edilemedi. RT-PCR ve PCR aracılığıyla hayvanların %6,74'ünde (13/193) solunum sistemi enfeksiyonu saptanırken, BRSV, BPIV-3, BVDV ve BoHV-1 prevalansı sırasıyla %2,59 (5/193); %0,52 (1/193); %2,59 (5/193) ve %1,04 (2/193) olarak tespit edildi. Elde edilen PCR ürünlerinin dizi analizleri ve sonrasında filogenetik analizleri yapıldı. Bunun sonucunda DO-7, ET-63 ve ET-124 kodlu dizinlerin BRSV'nin F gen bölgesine göre birbirleriyle yakın ilişkili olduğu; ET-81 kodlu dizinin HN proteini gen bölgesine göre BPIV-3c genotipinde; DO-48, ET-64 ve ET-94 kodlu dizinlerin 5'UTR gen bölgesine göre sırasıyla BVDV-1f, BVDV-1a ve BVDV-1l alt gruplarında; ET-121 kodlu dizinin ise gC gen gölgesine göre BoHV-1.1 alt grubunda yer aldığı belirlendi. Örneklerde çoklu enfeksiyon tespit edilemedi.The aim of this PhD thesis is to determine the presence of bovine respiratory syncytial virus (BRSV), bovine parainfluenza virus type 3 (BPIV-3), bovine viral diarrhea virus (BVDV) and bovine herpesvirus type 1 (BoHV-1), which are important viral pathogens of bovine respiratory disease. For that purpose, conventional virologic methods and molecular methods were employed. A total of 193 samples (133 nasal swabs and 60 lung tissue samples) from the cattle showing respiratory system findings were examined in Bursa and neighboring provinces. For virus isolation, all the samples were inoculated onto MDBK cell culture for 3 blind passages. In addition, the samples were tested by nested RT-PCR for BRSV; hemadsorption test and RT-PCR for BPIV-3; immunoperoxidase monolayer assay, antigen-ELISA and RT-PCR for BVDV; antigen-ELISA and PCR methods for BoHV-1. During virus isolation, 1 BPIV-3 (ET-81) and 2 non-cytopathogenic BVDV (DO-48 and ET-94) isolates were obtained. By antigen-ELISA tests, 10 samples were found to be positive for BVDV antigen, while there was no positive sample for BoHV-1 antigens. By RT-PCR and PCR, the prevalence of BRSV, BPIV-3, BVDV and BoHV-1 was determined as 2,59% (5/193); 0,52% (1/193); 2,59% (5/193) and 1,04% (2/193), respectively. 6,74% (13/193) of the animals tested were found to have respiratory infection caused by these viruses. Sequence analysis was performed on PCR products from selected samples. According to phylogenetic analyses, DO-7, ET-63 and ET-124 sequences were closely related to each other according to the F gene region of BRSV. ET-81 was found in the genotype of BPIV-3c according to HN protein gene region. DO-48, ET-64 and ET-94 sequences that were identified as BVDV-1 and 5′UTR-based analysis demonstrated the existence of BVDV-1f, BVDV-1a, and BVDV-1l subgenotypes, respectively. ET-121 sequence was found to take part in BoHV-1.1 subgroup according to gC gene region. No multiple infections could able to be detected in the samples

    Ankara Üniversitesi Veteriner Fakültesi

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    This study reports the high prevalence and molecular characterization of BoHV-1 infection in imported cattle with respiratory system disease after international transport. A high mortality rate of 14.16% (51/360) was reported in a group of animals imported from Hungary to Turkey in 2019. A total of 17 samples were evaluated (3 lung tissue and 14 nasal swab samples) from 15 cattle aged 6 to 9 months not vaccinated against BoHV-1. Virus isolation, polymerase chain reaction (PCR) and restriction endonuclease analysis (REA) procedures were performed within the scope of this study. By virus isolation in MDBK cells, cytopathologic effects was detected in 8 samples (3 lung tissue and 5 nasal swabs samples). The same eight samples were also found positive by BoHV-1 PCR targeting gC (UL44) gene region. According to the sequencing result, the sample (ID: 10054) dropped into a cluster of BoHV-1.1. The REA was applied to the samples to confirm the results of phylogenetic analysis. All of the isolates were identified in the subgroup BoHV-1.1 by REA. These results showed a high mortality risk for imported animals and the possibility for BoHV-1 entering the receiving country via imported animals after transport. This event is a serious problem both for the control of BoHV-1 as well as for animal health and welfare.Trdizi

    Going Beyond Counting First Authors in Author Co-citation Analysis

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    The present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
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