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研究生:劉子琦
研究生(外文):Tzu-Chi Liu
論文名稱:豬生殖與呼吸道綜合症病毒對吞噬細胞清除副豬嗜血桿菌功能影響之評估
論文名稱(外文):Evaluate the Effect of Porcine Reproductive and Respiratory Syndrome Virus Infection on Phagocytes to the Clearance of Haemophilus Parasuis
指導教授:李維誠李維誠引用關係
口試委員:羅登源簡茂盛
口試日期:2017-06-29
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:2017
畢業學年度:105
語文別:中文
論文頁數:62
中文關鍵詞:豬生殖與呼吸道綜合症副豬嗜血桿菌多形核白血球豬肺泡巨噬細胞
外文關鍵詞:Porcine reproductive and respiratory syndromeHaemophilus parasuisPolymorphonuclear leucocytePorcine alveolar macrophage
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豬生殖與呼吸道綜合症病毒(Porcine reproductive and respiratory syndrome virus, PRRSV)為保育階段豬隻最主要的呼吸道疾病之原發性病原,並常與Haemophilus parasuis (Hps)所引起之多發性漿膜炎混合感染。本試驗旨在評估PRRSV感染對豬隻吞噬細胞清除Hps的能力的影響。在試驗一中,以PRRSV對6週齡商用豬隻進行攻毒,在感染後不同時間點採血分離多形核白血球(Polymorphonuclear leucocyte, PMN),分析PMN對Hps吞噬及H2O2產生能力的影響。結果顯示在商用豬隻被PRRSV感染後,對白血球數量、吞噬功能及H2O2產生能力等並無明顯影響。在試驗二中,以in vitro方式評估不同毒力的PRRSV毒株感染Porcine alveolar macrophage (PAM)細胞後,對PAM吞噬Hps功能的影響。結果顯示在PRRSV感染後12~18小時間,PAM細胞的死亡 (Propidium iodide染色陽性)大幅度地升高(p<0.05)。相較於弱毒株感染,毒力較高的PRRSV毒株對PAM有較高的感染率,且會導致更多PAM的凋亡(p<0.05),且於感染後15及18小時吞噬及分泌過氧化氫的能力均顯著的降低(p<0.05)。根據以上實驗,毒力較高的PRRSV在PAM的複製速率較快,並對PAM功能造成更嚴重地抑制,致Hps能有更多的機會存活及進入血行感染。在活體上雖對PMN功能未見顯著影響,但Hps藉由規避補體調理及多數離乳後豬隻可能缺乏Hps特異IgG調理機制,使Hps能有更多的機會在血液中存活。這可能部分解釋多發性漿膜炎與PRRSV感染常同時被觀察到,尤其在缺乏Hps特異IgG抗體的豬隻中。
Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most popular respiratory pathogens in nursery pigs and frequently concurrent infection with Hemophilus parasuis (Hps). The purpose of this experiment was to evaluate the effect of PRRSV infection on the phagocytes to Hps infection. In the first experiment, 6 weeks old commercial pigs were challenged with PRRSV. After challenge, polymorphonuclear leukocytes (PMN) in blood at different time points were isolated for phagocytic assay. There were no significant changes on amount of leukocytes and the phagocytic activity and the production of hydrogen peroxide of PMN to Hps in PRRSV infected pigs, compared to negative control. In the second experiment, we evaluated the effect of a comparable virulence strain of PRRSV on porcine alveolar macrophage (PAM) function to Hps infection in vitro, compared to a low virulent strain. The results show that the percentages of dead PAMs , positive of propidium iodide staining, at 12 and 18 hours post infection in both PRRSV inoculated groups were significantly increased (p<0.05). Compared to the low virulent PRRSV infection, there were significantly increasing the positive numbers of PAM with PRRSV infection and dead PAM in the comparable virulent PRRSV infection (p<0.05). Meanwhile, both the phagocytic ability and the generation of hydrogen peroxide of PAM were significantly decreased (p<0.05) at 15 and 18 hours post PRRSV infection, especially in PAM infected with the comparable virulent PRRSV. In summary, the comparably virulent PRRSV shows greater replication rate and severely impairs PAM functions, leading more opportunity for Hps survival and invasion. Though there is no significant effect on PMN phagocytic function after PRRSV infection ex vivo, evading complement mediated opsonization in some virulent Hps and shortage of Hps specific IgG in most weaned pigs may also open another opportunity for Hps survival in blood. It may partially explain the high concurrent infections of polyserositis and PRRSV infection, especially in those pigs without Hps specific IgG antibodies.
摘要 -------------------------------------------------------------------------------- i
Abstract -------------------------------------------------------------------------------- ii
目錄 -------------------------------------------------------------------------------------- iv
表次 -------------------------------------------------------------------------------------- vi
圖次 -------------------------------------------------------------------------------------- vii
第一章 文獻探討 --------------------------------------------------------------------- 1
第一節 豬生殖及呼吸道綜合症 ----------------------------------------------- 1
1.1.1. 歷史背景 ----------------------------------------------------------- 1
1.1.2. 豬生殖及呼吸道綜合症之病毒特性 -------------------------- 2
1.1.3. 豬生殖及呼吸道綜合症之臨床症狀與病理變化 ----------------- 2
1.1.4. 豬生殖及呼吸道綜合症之致病機轉 -------------------------- 3
1.1.5. 豬生殖及呼吸道綜合症與宿主免疫反應的關係 ----------- 4
第二節 副豬嗜血桿菌 --------------------------------------------------------- 5
1.2.1. 背景資料 ---------------------------------------------------------- 5
1.2.2. 副豬嗜血桿菌之特性及分類 ---------------------------------- 5
1.2.3. 副豬嗜血桿菌感染之臨床症狀與病理變化 ---------------------- 6
1.2.4. 副豬嗜血桿菌感染之致病機轉 ------------------------------- 7
第三節 豬生殖及呼吸道綜合症與副豬嗜血桿菌之關係 --------------- 9
1.3.1. 豬隻呼吸系統之對細菌之防禦機制 ------------------------- 9
1.3.2. 豬生殖及呼吸道綜合症與副豬嗜血桿菌的關係 ---------- 10
1.3.3. 豬生殖及呼吸道綜合症與副豬嗜血桿菌的控制 ---------- 10
第二章 豬隻感染豬生殖及呼吸道綜合症病毒後對血液中嗜中性球清除豬
副嗜血桿菌功能之影響 --------------------------------------------------- 12
第一節 前言 ----------------------------------------------------------------------- 12
第二節 實驗目的與設計 ------------------------------------------------------ 14
第三節 材料方法 --------------------------------------------------------------- 15
第四節 結果 --------------------------------------------------------------------- 19
第五節 討論 --------------------------------------------------------------------- 21
第三章 不同毒力之豬生殖及呼吸道綜合症病毒感染對豬肺泡巨噬細胞功
能之影響 --------------------------------------------------------------------- 31
第一節 前言 ----------------------------------------------------------------------- 31
第二節 實驗目的與設計 ------------------------------------------------------ 33
第三節 材料方法 --------------------------------------------------------------- 34
第四節 結果 --------------------------------------------------------------------- 37
第五節 討論 --------------------------------------------------------------------- 39
第四章 參考文獻-------------------------------------------------------------------------- 49
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