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Öğe Characterization of Infectious Laryngotracheitis Virus Isolates from Turkey by Molecular and Sequence Analysis(Univ Agriculture, Fac Veterinary Science, 2020) Can-Sahna, Kezban; Abaylı, Hasan; Özbek, Remziye; Tonbak, Sukm; Bulut, HakanInfectious laryngotracheitis (ILT) is an economically important respiratory disease affecting the poultry industry worldwide. The aim of this study was to characterize the Turkish ILT virus (ILTV) isolates by sequencing and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). ILTV isolates obtained from laying hens in Turkey in 2003 and 2018 were used in this study. The isolates were analyzed for ICP4, gG, gE and TK gene regions by PCR. The amplification products were used in RFLP analysis to determine the differences among the isolates. Sequence analyses of TK and ICP4 regions were carried out and a phylogenetic tree was formed by using the Maximum Likelihood method. Nucleotide identity values were then calculated among five isolates and other strains/isolates in Genbank. In addition, about 200 amino acid sequences of the start and end regions of the ICP4 gene were compared to other strains in Genbank. PCR-RFLP analysis indicated that Turkish ILTV isolates were low-virulent. In general, the nucleotide sequence similarities of the TK and ICP4 gene regions among Turkish isolates and others was more than 95% (lower in some Egyptian and Bangladeshi strains, 41 and 45% respectively); in the amino acid sequence, it was close to 100%. As a result, PCR-RFLP results were similar in many gene regions. However, evolutionary analysis of ICP4 and TK gene regions did not yield reliable results based on geographic distribution or pathogenicity levels. For this purpose, different methods, such as Bayesian analysis or the involvement of samples from different gene regions can yield more reliable results, just like whole-genome sequences. (C) 2020 PVJ. All rights reservedÖğe Detection and molecular characterisation of feline viruses from swab samples(Akademiai Kiado Zrt, 2021) Abaylı, Hasan; Can Şahna, Kezban; Özbek, Remziye; Aslan, Öznur; Tonbak, Şükrü; Bulut, HakanFeline calicivirus (FCV), feline alphaherpesvirus 1 (FHV-1) and feline panleukopenia virus (FPLV) as well as retroviral agents such as feline leukaemia virus (FeLV) and feline immunodeficiency virus (FIV) are important viral pathogens of cats. The aim of this study was to detect and characterise FHV-1, FPLV, FeLV, FIV and feline foamy virus (FFV) in oropharyngeal, nasal and conjunctival swabs from 93 cats that had been screened for FCV previously. We wanted to determine the possible risk factors for infection with these viruses. The prevalence was found to be 12.9% for FHV-1 and 9.7% for FPLV. FIV was detected only in two samples and FeLV in one sample, whereas the presence of FFV was not demonstrated in any of the clinical samples. The statistical analysis of the results showed that breed, age, health status, and lifestyle are important predisposing factors to FHV-1 (P < 0.05). For FPLV, only clinically unhealthy animals were found to be at risk (P < 0.001). Sequence analysis revealed that the two FIV-positive samples in this study contained different (A and B) subtypes of the virus. This is the first report on the occurrence of subtype A FIV in Turkey.Öğe GENETIC DIVERSITY of OVINE HERPESVIRUS 2 STRAINS OBTAINED FROM MALIGNANT CATARRHAL FEVER CASES in EASTERN TURKEY(Elsevier, 2020) Turan, Turhan; Işıdan, Hakan; Atasoy, Mustafa Ozan; Sözdutmaz, İbrahim; Bulut, HakanMalignant Catarrhal Fever (MCF) is a generalized, definitive lethal disease affecting the epithelial and lymphoid tissues of the respiratory and digestive tract, mainly cattle and some wild ruminants such as deer, buffalo or antelope. The sheep-related form of MCF is known to be present in Turkey and is caused by ovine herpesvirus 2 (OvHV-2). The aim of this study was to reveal the genetic diversity of OvHV-2 strains obtained from MCF cases in Eastern Turkey where the livestock industry has an important impact on economic activities. For this purpose, RTA (Replication and transcription activator), FGARAT (formylglycineamide ribotide amidotransferase) and some of glycoprotein genes (0v7, Ov8 ex2, ORF27 and Ov9.5) were investigated in blood samples from 24 cattles, clinically diagnosed with MCF. Genomic data of chosen samples were furthermore used to characterize and undergo combined phylogenetic analysis to determine possible alleles and subvariants. The results showed that high level of OvHV-2 diversity existed in selected genes and strains carrying allelic variants might circulate both in two geographically distinct regions and in a region itself. Moreover, three different OvHV-2 types and various subtypes were identified based on multi locus approach. This study provides important data to epidemiological research and thereby helps to determine the source of the virus and understand the spread of the disease.Öğe Increased Circulatory Extrarenal 1 alpha,25-Dihydroxyvitamin D3 in Bilaterally Nephrectomized Rats(Bentham Science Publ Ltd, 2020) Dabak, Murat; Dabak, Durrin O.; Kuloğlu, Tuncay; Baydar, Ersoy; Bulut, Hakan; Özöner, ÖzgürBackground: Extrarenal 1 alpha,25-dihydroxyvitamin D-3 (1,25-D) locally produced by immune cells plays crucial roles in the regulation of the immune system. However, in vivo status of extrarenal 1,25-D and 25-hydroxyvitamin D (25-D) in acute inflammatory conditions are unknown. Objective: The aim of this study was to determine the extrarenal 1,25-D level in circulation in bilaterally nephrectomized rats, induced by low-dose lipopolysaccharide (LPS). Methods: Renal 1,25-D synthesis was terminated through bilateral nephrectomy in rats. The rats received intraperitoneal LPS (50 mu g/kg BW) three times and the experiment was ended 24 hours after nephrectomy. Serum 1,25-D, 25-D, calcium, phosphorus, intact parathyroid hormone, and calcitonin levels were measured and immunohistochemistry was applied to detect the sources of extrarenal 1,25-D synthesis. Results: Circulatory 1,25-D concentration remarkably increased in both LPS-treated and non-treated bilaterally nephrectomized rats. Elevated circulatory 1,25-D did not have hypercalcemic endocrinal effects. The increased 1,25-D level also resulted in a concurrent rapid and dramatic depletion of circulatory 25-D. Conclusion: Extrarenal 1,25-D could enter into the systemic circulation and, therefore, might have systemic effects besides its autocrine and paracrine functions.Öğe Molecular epidemiology of peste des petits ruminants cases associated with abortion in sheep and goat in Marmara Region of Turkey, 2018(University of Agriculture, 2020) Pestil, Zuleyha; Sait, Ahmet; Sayı, Orbay; Ozbaser, Fatma Tulin; Bulut, HakanThe aim of this study was to investigate the molecular epidemiology of peste des petits ruminants (PPR) infections associated with abortion in sheep and goat samples from the Marmara region of Turkey during 2018. The study was carried out from 116 sheep and 26 goat abortion cases. PPR virus (PPRV) detection in these samples was performed using real-time RT-PCR (Q-RT-PCR). Then, sequence analysis was performed from PPRV positive samples. Q-RT-PCR results demonstrated that 12 (10.34%) out of 116 sheep abortion samples and 3 (11.53%) out of 26 goat abortion samples were positive for PPRV genome. The sequence results of RT-PCR positive products revealed that the viruses causing the cases belong to lineage IV. Furthermore, molecular analysis showed that present cases were not related to PPRV vaccine strains or its mutants. Marmara region, where this study was conducted, is a neighbour of European countries such as Bulgaria and Greece. The first PPR cases in Europe were reported from Bulgaria at the beginning of 2018 and subsequently, other cases also reported before are mentioned in the present study. This study provides valuable information to understand the epidemiology of recently emerged PPRV cases in Europe and Turkey. Furthermore, because of the prevalence of PPRV in abortion samples in this study, these results suggest that PPRV may be one of the possible etiologic agents of abortions in sheep and goat. However, for clarification of the relationship between abortion and PPRV, there is need more robust epidemiological data and experimental infection studies. ©2020 PVJ. All rights reservedÖğe Short-term humoral immune response of the pcDNA4-G plasmid expressing the bovine ephemeral fever virus G gene in BALB/c mice(Ist Zooprofilattico Sperimentale Abruzzo & Molise G Caporale-Izs A&M, 2021) Abaylı, Hasan; Bulut, HakanBovine ephemeral fever (BEF) is an arthropod-borne viral disease characterised by a short-term clinical expression that can lead to significant losses in high-yielding cattle and water buffaloes. In this study, we aimed to generate a recombinant plasmid expressing the glycoprotein (G) of the BEF virus (BEFV) and to stimulate a humoral immune response to this protein in BALB / c mice immunised with the recombinant plasmid. Expression of the encoded protein was demonstrated by western blotting and immunoperoxidase tests. The suitable plasmids were intramuscularly administered to BALB/c mice on days 0, 14 and 21. The antibody response in the immunised mice was measured by a plaque reduction neutralization test (PRNT) and enzyme-linked immunosorbent assay (ELISA). According to BEFV ELISA, only two of the seven animals in these groups exceeded the cut-off value. A significant difference was observed in the mean OD values at 450 nm absorbance in the pcDNA4-G-immunised group when compared with those in the plasmid control group at 30 days (p < 0.05). According to PRNT50 results, a 1:20 (p < 0.05) antibody response was obtained at 30 days in pcDNA4-G (100 mu g)-immunised mice, whereas this ratio was 1:80 (p < 0.001) in BEFV-immunised mice (1,000 PFU/0.5 ml). We conclude that the humoral immune response was stimulated in experimental mice immunised with the recombinant plasmid. However, disappointingly, the antibody response was markedly low in pcDNA4-G-immunised mice.
