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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 王汎熒(Fun-In Wang) | |
| dc.contributor.author | Hao-Che Yen | en |
| dc.contributor.author | 顏浩哲 | zh_TW |
| dc.date.accessioned | 2021-07-11T15:46:49Z | - |
| dc.date.available | 2023-08-09 | |
| dc.date.copyright | 2018-08-09 | |
| dc.date.issued | 2018 | |
| dc.date.submitted | 2018-08-06 | |
| dc.identifier.citation | Akita, G.Y., Ianconescu, M., MacLachlan, N.J., Osburn, B.I., 1994. Bluetongue disease in dogs associated with contaminated vaccine. Vet Rec 134, 283-284.
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Oxford University Press Southern Africa, Cape Town, South Africa, 1201–1220 pp. Wilson, W.C., Ma, H.C., Venter, E.H., van Djik, A.A., Seal, B.S., Mecham, J.O., 2000. Phylogenetic relationships of bluetongue viruses based on gene S7. Virus Res 67, 141-151. Wirblich, C., Bhattacharya, B., Roy, P., 2006. Nonstructural protein 3 of bluetongue virus assists virus release by recruiting ESCRT-I protein Tsg101. J Virol 80, 460-473. Wouda, W., Roumen, M.P., Peperkamp, N.H., Vos, J.H., van Garderen, E., Muskens, J., 2008. Hydranencephaly in calves following the bluetongue serotype 8 epidemic in the Netherlands. Vet Rec 162, 422-423. Zhang, N., MacLachlan, N.J., Bonneau, K.R., Zhu, J., Li, Z., Zhang, K., Zhang, F., Xia, L., Xiang, W., 1999. Identification of seven serotypes of bluetongue virus from the People's Republic of China. Vet Rec 145, 427-429. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/79135 | - |
| dc.description.abstract | 藍舌病(BT)是經由庫蠓傳播的反芻動物病毒性疾病，對經濟及野生反芻獸皆有感受性，進而造成全世界畜牧業嚴重的經濟損失。過去的研究指出藍舌病病毒(BTV)在台灣的反芻獸通常呈現無症狀的亞臨床感染，並分別在金門和屏東地區分離出2株病毒。基因組分析顯示這2個分離株在segment 7上出現了正向選擇(positive selection)。基因segment 7轉譯成最外圍的核心蛋白質VP7，是在病毒進入昆蟲細胞時不可或缺的蛋白質之一，這些證據顯示BTV可能會為了適應台灣的環境而演化。自從18000年前冰河時期結束以來，台灣島便與其他亞洲陸塊分離，島上生物因而演化出許多特有種，其中也包括了數種有機會傳播BTV的庫蠓屬昆蟲。本研究的假說是台灣特有的庫蠓屬昆蟲在傳播藍舌病時提供了BTV基因segment 7演化壓力。研究目的在於調查目前在台灣島流行的BTV病毒株及其分佈。以競爭型酵素結合免疫吸附法檢測在台灣各地區採集的牛羊血清中是否有BTV-VP7特異性抗體，結果顯示牛的血清陽性率為26.7％，山羊為18.6％。以RT-PCR檢測牛羊抗凝全血樣本中的BTV-VP1核酸訊號，其中山羊檢體中有9個陽性樣本，而使用OIE推薦之BTV-NS1引子，所有檢體均呈現陰性。為了分離病毒，將數個核酸陽性的全血樣本接種至雞胚胎和BHK21細胞株，但未能分離到新的病毒株。本研究中亦比較了台灣、日本、中國、印度、澳大利亞等鄰近地區BTV病毒株的VP7胺基酸序列，發現在台灣及日本株有3個特定位置的氨基酸與其他地區分離株不同，這些變異位於VP7三聚體結構上的相鄰位置，而且是台灣株BTV2和BTV12以及日本株獨有的，這個發現揭示了BTV在鄰近地區間演化的過程中，為了適應當地庫蠓時VP7可能扮演的角色。 | zh_TW |
| dc.description.abstract | Bluetongue (BTV) is an arthropod-borne disease, in domestic and wild ruminants, caused by bluetongue virus (BTV), and leads to great economic loss worldwide. Previous studies showed BTV infection in Taiwan was often subclinical, and 2 viruses had been isolated from specimens in Kinmen and Pingtung districts, respectively. Genetic analysis of the 2 isolates showed a positive selection of segment 7, suggested that BTV might evolve and adapt when entering Taiwan as a new environment. Genome segment 7 encodes an outermost core protein, VP7, which is the most accessible protein of the BTV core that may participate in cell entry. Taiwan is geographically isolated for a long period of time since the end of last ice age 18000 years ago, and inhabited by several endemic vector species, including Culicoides, the vector of BTV. The hypothesis is that these endemic Culicoides species in Taiwan provide an evolution pressure to BTV, especially on genome segment 7. The aim of the project was to investigate the new viral strains and the distribution of these strains on Taiwan. Blood samples were collected from different districts in Taiwan. ELISA tests were performed for anti-BTV group specific antibody revealing seropositive rates of 26.7% in cattle by heads, and 18.6% in goats. BTV nucleic acid in whole blood was detected via RT-PCR targeted on VP1 segment revealed 9 positive samples from goat. Detection via RT-PCR target on NS1segment revealed negative in all samples. A few RT-PCR positive blood samples were selected for viral isolation via inoculation into chicken embryos and BHK21 cell line but failed to isolate new virus strain. Sequence alignment of Taiwan BTV isolates VP7 with isolates from neighboring areas including Japan, China, India and Australia showed 3 specific amino acids, that are unique to Taiwan BTV2 and BTV12 and Japan isolates, located close by on the three-dimensional model of VP7 trimer, suggesting the role of VP7 in BTV evolution in local Culicoides vector species in these geographically neighboring areas. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-11T15:46:49Z (GMT). No. of bitstreams: 1 ntu-107-R04644003-1.pdf: 2580297 bytes, checksum: c22c94752dd7b673d9819c39488fe12b (MD5) Previous issue date: 2018 | en |
| dc.description.tableofcontents | Certificate i
Acknowledgements ii 中文摘要 iii Abstract iv Contents v 1. Introduction 1 2. Literature review 2 2.1 Bluetongue disease 2 2.1.1 History 2 2.1.2 Host range 3 2.1.3 Clinical signs 4 2.1.4 Pathogenesis 5 2.1.5 Gross lesions 6 2.1.6 Histopathologic lesions 7 2.2 Bluetongue virus 8 2.2.1 Genome 8 2.3 Structures and functions of viral proteins 8 2.3.1 The major core proteins: VP3 and VP7 9 2.3.2 The minor core proteins: VP1, VP4 and VP6 9 2.3.3 The outer structural proteins: VP2 and VP5 10 2.3.4 The non-structural proteins: NS1, NS2, NS3, NS3A and NS4 11 2.4 Viral replication 11 2.4.1 Virus entry 11 2.4.2 Replication of viral RNA 12 2.4.3 Assembly of progeny virus 12 2.4.4 Virus release 13 2.5 Vectors of bluetongue 13 2.5.1 Culicoides biting midges 13 2.5.2 Culicoides as vector 14 2.5.3 Taiwan Culicoides Species 15 2.6 Bluetongue disease in Taiwan 15 3. Materials and Methods 15 3.1 Experimental design 15 3.2 Sample collection 16 3.3 Sample preparation 16 3.4 Bluetongue-specific antibody testing 17 3.5 Sample RNA extraction 17 3.6 Reverse transcription polymerase chain reaction (RT-PCR) for field sample survey, targeting BTV VP1, VP7 and NS3 18 3.7 Gel electrophoresis 19 3.8 Real-time RT-PCR (qRT-PCR) for field sample survey targeting BTV VP1 19 3.9 Nucleotide Sequencing: 19 3.10 Inoculation of embryonating chicken eggs (ECE) 20 3.11 Cell culture inoculation 21 3.12 BTV-VP7 Amino acid alignment of Asian BTV isolates 22 4. Results 22 4.1 Seroprevalence of BTV in cattle was higher than goats in Taiwan. (Tables 2-3; Figure 2) 22 4.2 BTV-VP1 nucleic acid was detected via RT-PCR in blood samples from goats (Figure 3) 23 4.3 BTV-VP1 nucleic acid was detected via qRT-PCR in blood samples from goats (Figures 4 -5) 23 4.4 ECE inoculated with sample No. G534 and No. 1136 showed stunted growth with lighter body weight. (Figure 6; Table 4) 23 4.5 No CPE was observed in BHK-21 inoculated with ECE tissue emulsion of sample No. G534 and sample No. 1136 (Figure 7) 24 4.6 Analysis of BTV-VP7 Amino acid sequence alignment of BTV2/KM/2003, BTV/PT/2003 and BTV isolates in neighboring areas showed 3 specific amino acid differences (Table 5; Figure 8) 24 5. Discussion 25 References 29 Figure 1. Taiwan island is divided into 4 areas based on the latitudes, the geographic relationship to the Central Mountain Range, and the geographic location of the districts. 39 Figure 2. Frequency distribution of the seroprevalence rates (herds/flocks) of the bluetongue virus infection in all collected samples in Taiwan (2016-2017). 40 Figure 3. One-step RT-PCR yields specific amplicons of 94 base pairs using BTV-VP1 specific primers. 41 Figure 4. Melting curve and peak analysis for BTV-VP1 in goat blood samples. 42 Figure 5. Melting curve and peak analysis for BTV-VP1 in goat blood samples with higher fluorescence and Cp value (No. G534 and No. 1136). 43 Figure 6. ECEs inoculated with BTV2/KM/2003, field RBC samples, and negative controls, respectively. 44 Figure 7. BHK-21 cells inoculated with tissue emulsions of BTV2/KM/2003, samples No. G534 and No. 1136, and negative inoculated chicken embryos. 45 Figure 8. Three-dimensional model of VP7 trimers showing 3 specific amino acids on Taiwan and Japan isolates are consistently different from those isolates of the surrounding countries. Refer also to Table 5. The 3 specific sites are No. 82, Arginine; No. 328, Aspartate; and No. 336, Glutamine. Even though No. 82 locates far away from No. 328 and No.336 in the linear sequence, they are associated closely on the surface of VP7 on the three-dimensional model. 46 Table 1. Primer sequences for the detection of viral genome in field samples by one step RT-PCR and qRT-PCR 47 Table 2. Seroprevalence of BTV infection in cattle in Taiwan (2016-2017) 48 Table 3. Seroprevalence of BTV infection in goats in Taiwan (2016-2017) 49 Table 4. Body weight of chicken embryo inoculated with field samples at 7 DPI 50 Table 5. VP7 Amino acid alignment of BTV2/KM/2003 and BTV isolates in neighboring areas 51 | |
| dc.language.iso | en | |
| dc.subject | 藍舌病 | zh_TW |
| dc.subject | 基因組序列比對 | zh_TW |
| dc.subject | 第七病毒蛋白 | zh_TW |
| dc.subject | 第七段基因組 | zh_TW |
| dc.subject | genome alignment | en |
| dc.subject | VP7 | en |
| dc.subject | bluetongue | en |
| dc.subject | segment 7 | en |
| dc.title | 臺灣藍舌病病毒segment 7基因組及胺基酸之序列分析 | zh_TW |
| dc.title | Analysis of Genome Segment 7 and Amino Acid Sequence of Bluetongue Virus in Taiwan | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 106-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 李璠,李龍湖,許天來,張家宜 | |
| dc.subject.keyword | 藍舌病,第七段基因組,第七病毒蛋白,基因組序列比對, | zh_TW |
| dc.subject.keyword | bluetongue,segment 7,VP7,genome alignment, | en |
| dc.relation.page | 56 | |
| dc.identifier.doi | 10.6342/NTU201802556 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2018-08-06 | |
| dc.contributor.author-college | 獸醫專業學院 | zh_TW |
| dc.contributor.author-dept | 分子暨比較病理生物學研究所 | zh_TW |
| dc.date.embargo-lift | 2023-08-09 | - |
| 顯示於系所單位： | 分子暨比較病理生物學研究所 | |
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