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研究生:李苡菡
研究生(外文):Yi-Han Li
論文名稱:台灣中部三種不同PRRS感染類型豬場之監控
論文名稱(外文):Monitoring Porcine Reproductive and Respiratory Syndrome Virus Infectious Status in Three Different PRRS-endemic Farms in Central Taiwan
指導教授:李維誠李維誠引用關係
口試委員:簡茂盛邱明堂羅登源
口試日期:2016-06-20
學位類別:碩士
校院名稱:國立中興大學
系所名稱:獸醫病理生物學研究所
學門:獸醫學門
學類:獸醫學類
論文種類:學術論文
論文出版年:2016
畢業學年度:104
語文別:中文
論文頁數:65
中文關鍵詞:豬生殖與呼吸綜合症PRRS血清抗體ORF5親緣樹狀圖分析
外文關鍵詞:PRRSVPRRSV antibody profilesORF5 phylogenetic analysis
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豬生殖與呼吸綜合症病毒(PRRSV)廣泛存在台灣多數養豬場,此病原對牧場之感染程度受到諸多因素影響,包括病毒毒力、毒株、管理方式與生物安全措施等。本試驗自2014年9月起至2015年底,近一年半期間,追蹤台灣中部三個大型一貫式養豬場(HB、SJ與YM牧場)之不同年齡層族群採血進行PRRSV血清抗體、病毒血症及ORF5核酸變異程度與親緣樹狀圖分析,合併牧場生物安全與管理方式等因素,分析其對牧場之感染壓力的影響。HB牧場於監控期間之血清混合樣品平均核酸檢出率較低,核酸序列相似性高,且保育舍維持穩定高育成率,顯示具良好外在生物安全措施及管理模式的牧場,得以避免外源性病毒株入侵,並維持長期穩定感染狀態。SJ牧場於2013-2015年發生至少三起母豬流產與離乳前仔豬死亡率增加之PRRS疫情,並於監控期間檢出3種異源株。由牧場資料可知,病原由鄰近多個豬場經空氣傳播進入、或由PRRS汙染場購入公豬飼養與配種皆為引入異源株之可能危險因子。因此,防止新毒株再次進入牧場為其重要防治考量。反觀YM牧場,監控期間雖無明顯異源株入侵,但保育舍豬隻長期受嚴重PRRSV相關之呼吸道疾病與高死亡率所困擾,此與牧場毒株毒力偏強、病毒於豬群持續循環感染,及不適當的生物安全措施與管理方式,皆致使PRRS難以受到控制。因此,改善生物安全措施與管理策略,配合疫苗使用應可強化YM牧場疾病控制效果。整體而言,由於PRRSV異源株間之交叉保護力可能不足,對台灣牧場而言,在有或無使用疫苗情況下,生物安全措施之提升以避免野外毒株進入牧場,及改善管理方式降低環境病毒感染壓力為目前台灣畜殖場不可忽視之防治策略。

Porcine reproductive and respiratory syndrome virus (PRRSV) infection has been endemic in most pig farms in Taiwan. The impact on PRRSV infection in endemic farms is dependent on various factors including virulence, strain, managerial factors and biosecurity. The aim of this study was to monitor PRRSV antibody profiles, viremia, divergence of PRRSV ORF5 phylogenetic analysis and the implement of biosecurity and managerial factors in three farrow-to-finish farms (Farm HB, SJ and YM) in central Taiwan from the middle of 2014 to 2015, which were correlated with the pressure of PRRSV infection within the farm. Blood samples were regularly collected from different age groups in these 3 farms. Farm HB displayed a relative stable PRRSV infection with low frequency of PRRSV viremia in pooled samples and strains with highly genetic identity which were correlated with good external biosecurity, management and good performance in nursery pigs. Due to shortage of external biosecurity, Farm SJ showed at least 3 outbreaks of abortions and increasing mortality before weaning between 2013 and 2015, which were correlated with the 3 different exogenous PRRSV strains invasions. Airborne transmission from several neighboring pig sites and boars purchased from PRRSV-positive farm might be the potential risk factor for these outbreaks. Therefore, to avoid introduction of exogenous PRRSV into the herd is a critical PRRS control issue for Farm SJ. In contrast, Farm YM has been suffered from severe PRRSV-associated respiratory diseases with high mortality in nursery unit for a long period, even though there was no obvious exogenous PRRSV invasion detected during monitoring period. However, the virulent PRRS virus strain persistently circulated in the farm and the improper biosecurity and management might contribute the difficulty for PRRS control. Therefore, implementation of biosecurity, managerial improvement and vaccination may have optimistic effects for disease control for Farm YM. In conclusion, as the insufficient cross protection against heterologous PRRSV, the most important of all, including the implementation of biosecurity to avoid exogenous virus invasion and improving management and facility with/without vaccination to decrease infectious pressure in herds are currently key strategies for PRRSV control in Taiwan.

摘要 -----------------------------------------------i
Abstract ---------------------------------------------ii
目錄 -------------------------------------------------iv
表次 --------------------------------------------------v
圖次 -------------------------------------------------vi
第一章 文獻探討 ---------------------------------------1
第一節 豬生殖與呼吸綜合症歷史背景 -----------------------1
第二節 豬生殖與呼吸綜合症病毒與疾病 ----------------------2
1.2.1. 病毒特性 ------------------------------------2
1.2.2. 傳播途徑 ------------------------------------3
1.2.3. 病毒變異特性與親緣分析 -----------------------6
1.2.4. 致病機制 ------------------------------------8
1.2.5. 臨床症狀與病理變化 ---------------------------9
1.2.6. 免疫反應 -----------------------------------10
1.2.7. PRRS防治策略與影響因子 ----------------------12
第二章 豬生殖與呼吸綜合症血清及分子生物學監控 ------------14
第一節 實驗動機 --------------------------------------14
第二節 材料與方法 -------------------------------------15
2.2.1. 牧場背景資料 ------------------------------------15
2.2.2. 血液採集與組織採樣 -------------------------------17
2.2.3. PRRSV血清特異性抗體定量-酶聯免疫吸附試驗(ELISA) ----17
2.2.4. 血清與組織乳劑PRRSV病毒核酸純化 -------------------18
2.2.5. PRRSV之ORF5與ORF7反轉錄聚合酶鏈鎖反應(reverse transcription polymerase chain reaction, RT-PCR)及巢式反轉錄聚合酶鏈鎖反應(nested RT-PCR, nRT-PCR) ----------------18
2.2.6. 各牧場分娩舍與保育舍死亡率/育成率紀錄 --------------20
2.2.7. YM一場外來豬輸入及育成資料記錄 --------------------20
2.2.8. ORF5核酸序列變異度判斷標準 -----------------------21
2.2.9. 牧場內外生物安全等級評估指標 ----------------------21
2.2.10. 統計分析 ---------------------------------------21
第三節 結果 ------------------------------------------22
2.3.1. HB牧場 -----------------------------------------22
2.3.2. SJ牧場 -----------------------------------------23
2.3.3. YM二場 -----------------------------------------24
2.3.4. YM一場外來豬輸入試驗與監控 ------------------------25
2.3.5. 牧場內外生物安全等級評估 --------------------------28
第四節 討論 ------------------------------------------31
第三章 參考文獻 --------------------------------------52


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