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研究生:林上海
研究生(外文):Lin Sun-Hi
論文名稱:甲魚出血性胃腸炎之病因學及組織病理學之研究
論文名稱(外文):Study on the etiology and histopathology of heamorrhagic gsatroenteritis in soft-shell turtle
指導教授:董明澄蔡信雄蔡信雄引用關係
指導教授(外文):MC TungSS Tsai
學位類別:碩士
校院名稱:國立屏東科技大學
系所名稱:獸醫學系
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:1999
畢業學年度:87
語文別:中文
論文頁數:96
中文關鍵詞:甲魚(中華鱉)親水性產氣單胞菌出血性胃腸炎
外文關鍵詞:Soft-Shell TurtleAeromonas hydrophilaHaemorrhagic gastroenteritis
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自1997年1月至1998年12月底止,台灣南區魚病中心甲魚送檢病例中,細菌性疾病共佔97例,而細菌性胃腸炎佔45例,高達46.39%。經細菌分離、鑑定後共得72株,本試驗以傳統生化鑑定配合Biolog 及API 20E 快速鑑定,結果Aeromonas hydrophila共得19株 (佔42.2%)。Aeromonas sobria 11株(佔24.4%)。其它尚有Aeromonas trota、 Aeromonas media-like及Plesiomonas。以平板稀釋法測定分離菌株對Nitrofurantoin (F/M)、Flumequine (UB)、Oxolinic acid (OA)、Sulfamethoxazole trimethoprim (SXT)、Nalidixic acid (NA)、Chloramphenicol (C)、Amoxicillin (Amc)、Kanamycin (K)、Tetracycline (Te)、Neomycin (N)、Cephalothin (KF)等11種抗菌劑之感受性,結果發現A. hydrophila對OA、UB及F/M感受性較高,Amc、Te及KF感受性最差。本試驗將分離之菌株A. hydrophila 做攻毒試驗。將試驗甲魚各分為8組,1組對照組,每組5隻,結果A. hydrophila經口攻毒,得到LD50為9.45×104 CFU/ml;腹腔攻毒,LD50為7.13×104 CFU/ml。其組織病理學變化為胃、十二指腸、小腸、大腸呈現粘膜層明顯充血,甚至出血,而粘膜下層水腫、血管明顯充血,產生之病變與野外發病甲魚之胃腸組織病理變化相同。經攻毒之甲魚於剖檢時進行細菌重分離胃腸分離細菌,也能從胃、十二指腸及大腸回收病原菌。經野外病例所分離到之病原,經攻毒後,除了可產生相同病變,且又可從供毒動物重分離到原攻毒之病菌,符合Koch*s定律,故證明A. hydrophila可誘發出血性胃腸炎。另外,本試驗有抽取本菌株之細胞外產物(Extracellular Product,ECP) ,將此細胞外產物以腹腔注射來測定其對甲魚之致病性。
From January of 1997 to December of 1998,total of 97 cases of soft-shell turtle infected with bacterial pathogens were collected in the Taiwan Southern Reginal Center of Fish Diseases. Forty five cases of them were associated with bacterial gastroenteritis and represented as 46.4% of the total bacterial cases.
Bacterial identifieation was carried out by the traditional biochemical tests alone with the combination of Biolog and API 20E rapid identification system.The results of aeromonads identification were as follows:there were 19 strains (42.2%) of Aeromonas hydrophila,11 strains (24.4%) of A. sobria, etc.
Eleven antimicrobiol agents for drugs sensitivity test were as follows:Nitrofurantoin (F/M)、Flumequine (UB)、Oxolinic acid (OA)、Sulfamethoxazole-Trimethoprim (SXT)、Nalidixic acid (NA)、Chloramphenicol (C)、Amoxicillin (Amc)、Kanamicin (K)、Tetracycline (Te)、Neomycin (N) and Cephalothin (KF) by Bauer-Kirby Test.
The results of Bauer-Kirby test for drugs sensitivity of 72 isolates were as follows:The most effective antibiotics were Oxolinic acid (OA)、Flumequine (UB) and Nitrofurantoin (F/M),while the most resistant ones were Amoxcillin (Amc)、Tetracycline (Te) and Cephalothin (KF).
The artificial infection was carried out by intraperitoneal inoculation (IP) and oral administration (Peros) of AR-98-368 strain of A. hydrophila. The median of lethal dose (LD50) of IP and peros were 7.13*104 CFU/ml and 9.45*104 CFU/ml respectively.
Histopathologically, serious mucosal hyperemia and hemorrhage with hyperemia and edema of submucosa was scattered through out the whole digestive tract, including oral cavity,stomach,denodenaum,small and large intestine.
Histopathological changes of digestive tract of artificially infected soft-shell turtle was similar to those of naturally infected cases. And the same bacterium of Aeromonas hydrophila, which was used for inoculation, was reisolated. These facts are satisfactory to the Koch愀 postulation.
目 錄
頁次
中文摘要……………………………………………… Ⅰ
英文摘要……………………………………………… Ⅲ
誌謝…………………………………………………… Ⅴ
表次…………………………………………………… Ⅸ
圖次…………………………………………………… Ⅹ
第一章 緒言……………………………………………1
第二章 文獻探討………………………………………3
第一節 中國鱉簡介…………………………………3
一、種類及分佈……………………………………3
二、中國鱉的生態習性……………………………3
三、中國鱉的食性…………………………………4
四、中國鱉之生理特性……………………………4
第二節 魚類胃腸炎回顧……………………………5
第三節 Aeromonads 的歷史沿革……………………7
一、Aeromonas hydrophila菌…………………………7
二、Aeromonas sobria菌………………………………8
第四節Aeromonads之特性……………………………9
一、Aeromonas hydrophila之特性……………………9
二、Aeromonas sobria之特性………………………10
第五節Aeromonads的致病性………………………10
一、Aeromonas hydrophila之致病性………………10
二、Aeromonas sobria及其他Aeromonas spp.
的致病性…………………………………………14
第六節Aeromonads對動物造成之組織病理學變化
及其致病機轉……………………………………15
一、組織病理學變化……………………………15
二、Aeromonas hydrophila之致病機轉……………17
第三章 材料與方法…………………………………20
第一節 流行病學調查………………………………20
第二節 細菌性出血性腸炎病原分離………………20
一、肉眼病理學檢查………………………………20
二、剖檢病理學檢查………………………………20
三、組織病理學觀察………………………………20
第三節 出血性胃腸炎病原分離……………………24
一、臟器細菌分離…………………………………24
二、胃腸內容物之濕壓片及抹片染色檢查………24
三、電子顯微鏡切片檢查…………………………25
第四節 細菌生化特性試驗…………………………26
一、細菌形態觀察………………………………27
二、生化性狀試驗………………………………27
三、細菌耐酸性試驗……………………………32
第五節 藥物感受性試驗…………………………33
一、細菌濃度的調整……………………………33
二、抗生素紙錠試驗……………………………33
第六節 人工感染試驗……………………………35
一、實驗材料……………………………………35
二、甲魚接種試驗………………………………35
(一)菌液腹腔接種………………………………35
(二)菌液經口接種………………………………35
第七節 組織病理學觀察…………………………37
第四章 結 果………………………………………38
第一節 流行病學調查……………………………38
第二節 剖檢肉眼病理檢查及組織病理學觀察……38
一、肉眼病理檢查…………………………………38
二、組織病理學檢查………………………………39
第三節 出血性胃腸炎病原分離……………………40
一、臟器細菌分離………………………………40
二、胃腸內容物之濕壓片及抹片染色檢查………40
三、胃腸道電顯檢查…………………………………40
第四節 生化性狀之鑑定結果………………………41
一、細菌形態觀察……………………………………41
二、生化性狀試驗結果………………………………41
三、細菌耐酸性試驗結果…………………………41
第五節 藥物感受性試驗……………………………42
第六節 甲魚人工接種感染結果………………………43
一、腹腔接種…………………………………43
二、口腔接種………………………………………44
第五章 討 論…………………………………………46
參 考 文 獻………………………………………54
圖 表………………………………………………68
附 錄…………………………………………………93
表 次
頁次
表一. 1997~1998年甲魚病歷分類…………………68
表二. 甲魚出血性胃腸炎細菌分離統計表…………69
表三. A. hydrophila細菌分離之生化性狀……………70
表四. A. sobria細菌分離之生化性狀………………71
表五. 甲魚胃腸道之正常pH值………………………72
表六. A. hydrophila藥物感受性試驗…………………74
表七. A. sobria藥物感受性試驗……………………75
表八. A. hydrophila AR-98- 368株腹腔接種死亡統
計………………………………………………76
表九. A. hydrophila以腹腔接種所引發之胃腸道
病變統計表……………………………………77
表十. A. hydrophila AR-98-368株口服接種死亡統計…78
表十一 A. hydrophila以經口接種所引發之胃腸病
變統計………………………………………79
圖 次
頁次
圖一:將Aeromonas hydrophila 革蘭氏染色,菌體為革蘭
氏陰性短桿菌(1000X)。……………… 80
圖二:野外發病之甲魚顯示精神沉鬱,活力下降,食慾減
退。……………………………… 80
圖三:野外發病甲魚,有時會有頸部腫大及腹部會稍膨大,
甚至蒼白。……………………………… 81
圖四:野外發病甲魚,剖檢時常發現腹腔積水、肝臟及胃腸
道漿膜有多發出血點到出血斑。………………… 81
圖五:野外發病甲魚之胃腸道粘膜面呈現充出血現象。……………… 82
圖六:野外發病甲魚之組織病理變化,胃呈現表淺層壞死,
粘膜固有層有炎症細胞浸潤。………………82
圖七:野外發病甲魚組織病理變化:小腸粘膜層出血、充血
,粘膜下層充血及水腫。………………83
圖八:野外發病甲魚肺部組織病理變化:充出血。………………83
圖九:野外發病甲魚肝臟組織病理變化:充出血。………………84
圖十:Aermonas hydrophila AR-98- 368株腹腔接種胃腸道漿
膜面潮紅。………………84
圖十一:A. hydrophila AR-98-368株腹腔注射:口腔常呈現
充出血。……………… 85
圖十二:A. hydrophila AR-98-368株腹腔注射:胃腸粘膜面
充出血。……………… 85
圖十三:A. hydrophila AR-98-368株腹腔接種之組織病理學
變化:口腔粘膜層,表淺層充出血、壞死。……………… 86
圖十四:A. hydrophila AR-98-368株腹腔接種之組織病理學變
化:胃組織病理切片下,粘膜層充血、粘膜下層充血
及水腫。……………………………… 86
圖十五:A. hydrophila AR-98-368株腹腔接種之組織病理學
變化:十二指腸表淺層壞死。粘膜下層充出血及水
腫。……………………………… 87
圖十六:A. hydrophila AR-98-368株腹腔接種之組織病理學
變化:小腸粘膜層充血、粘膜肌層充血、粘膜下層
水腫、肌肉層充血。……………… 87
圖十七:A. hydrophila AR-98-368株腹腔接種之組織病理學
變化:大腸粘膜層、粘膜肌層充血、粘膜下層充血
及水腫,漿膜層充血。……………… 88
圖十八:A. hydrophila AR-98-368株腹腔接種之組織病理學變
化:心臟出現嚴重出血現象。……………… 88
圖十九:A. hydrophila AR-98-368株腹腔接種之組織病理學變
化:腎臟有充血,絲球體尿酸鹽沉著,壞死。……………… 89
圖二十:A. hydrophila AR-98-368株腹腔接種之組織病理學變
化:肺臟有充出血。………………89
圖二十一:A. hydrophila AR-98-368株經口接種,予以ketamine
麻醉,再以胃導管連接注射針筒,將導管插至胃賁
門處,即可行攻毒。……………… 90
圖二十二:A. hydrophila 368經口接種,肉眼病變呈現腹腔積
水、肝臟及胃腸道充血。……………… 90
圖二十三:A. hydrophila 368經口接種。胃腸道粘膜充、出血。
……………………………… 91
圖二十四:A. hydrophila AR-98-368株經口接種之組織病理學
變化:胃組織病理切片下,粘膜層及粘膜下層出現
充血。……………… 91
圖二十五:A. hydrophila AR-98-368株經口接種之組織病理學
變化:十二指腸粘膜面充出血,粘膜層充出血及水
腫。……………… 92
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