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研究生:劉春田
研究生(外文):Chuen-Tyan Liou
論文名稱:台灣環頸雉盲腸型球蟲Eimeriacolchici之病原性及控制之探討
論文名稱(外文):Pathogenicity and Control of Eimeria colchici Infection in Taiwan Ring-necked Pheasant
指導教授:王俊秀王俊秀引用關係
指導教授(外文):Jiunn-Shiow Wang
學位類別:博士
校院名稱:國立中興大學
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:2001
畢業學年度:89
語文別:中文
論文頁數:147
中文關鍵詞:環頸雉環頸雉盲腸型球蟲化學藥劑氨水臭氧免疫接種病原性控制
外文關鍵詞:Phasianus colchicusEimeria colchiciChemical drugAmmonia solutionOzoneImmunizationPathogenicityControl
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中文摘要
自1997年7月至1999年3月,對臺灣地區19場較具規模的動物園和雉雞飼養場進行鶉雞類球蟲科寄生狀況調查,在41種395籠(件)之糞樣中,檢出球蟲卵囊96件(24.3%)。其中屬於Eimeria屬75件(78.1%)、Isospora屬24件(25.0%)、Gousseffia屬8件(8.3%)、Pythonella屬5件(5.2%),其中Gousseffia屬及Pythonella屬球蟲在國內禽類球蟲檢出尚屬首次。
由野外病例分離得到之Eimeria colchici,以單一卵囊增殖法,經人工接種繼代增殖方式取得足量之環頸雉盲腸型球蟲E. colchici未芽孢化卵囊,取其中部分卵囊按試驗組別等量分配後,各分別以13種化學藥劑測試卵囊之芽孢化比率,結果以Ammonia solution抑制卵囊之芽孢化作用效果最好,抑制效率可達92.85%,其次為Phenol有38.22%效果,再其次為Hyamine有18.73%之效果。另以人工接種方式同時取得之E. colchici卵囊,先予以芽孢化處理後,亦按試驗組別將卵囊等量分配後,分別與前述各種化學藥劑相同之濃度浸漬處理1小時,並以蒸餾水離心清洗5次後,再人工接種於14日齡環頸雉,每隻經口感染劑量均為120,000個卵囊,結果仍以Ammonia solution組之死亡率最低為16.67%(3/18),其餘各組(含陽性對照組)死亡率均高達100%(18/18)。
為建立氨水有效的作用濃度,經以人工接種增殖方式分別取得環頸雉盲腸型球蟲Eimeria colchici糞中及盲腸粘膜上之未芽孢化卵囊,各分別以0.75%、0.25%、0.125%及0.025% 氨水浸漬處理1小時後,然後以蒸餾水離心清洗4次,於28℃定溫培養36小時後,計算各氨水處理組之卵囊芽孢化比率。結果顯示氨水之濃度愈高,對於卵囊之芽孢化作用抑制效果愈好。自糞中收集得到之未芽孢化卵囊,經上述4種不同濃度之氨水浸漬處理後,依氨水濃度高低所得之芽孢化抑制效率依序分別為99.82%,37.14%,28.55%,0%;而自盲腸粘膜取得之未芽孢化卵囊,以同樣之濃度、時間處理結果,所得之芽孢化抑制效率依序分別為99.68%,96.42%,57.81%,26.05%,這顯示出已排出體外之未芽孢化卵囊,於0.25%氨水以下濃度時,比尚留在盲腸粘膜上之未芽孢化卵囊具抵抗性,且0.025% 氨水以下之濃度對已排出體外之卵囊並不具殺滅能力。
經以人工接種增殖方式取得尚未芽孢化之E. colchici卵囊,卵囊以蒸餾水充分懸浮後,按試驗組別等量分配,各組分別以臭氧曝氣處理15、30及60分,經重複試驗三次結果顯示,凡經臭氧曝氣處理後卵囊之芽孢化比率均較對照組為低,且曝氣處理時間愈久,芽孢化比率愈低。已芽孢化之E. colchici卵囊以相同方式處理後,經口接種於14日齡環頸雉,每隻各120,000個卵囊,結果亦顯示,經臭氧曝氣處理時間愈久者,鳥隻死亡率愈低,且每克糞材所排出之卵囊數亦較對照組少。
此外,以人工接種繼代增殖保存株之E. colchici卵囊,接種14日齡雛雉,免疫組及對照組各為28隻,免疫組(五組)整個均屬低劑量,各免疫組每隻分別經口接種100、250、500、1,000及2,000個芽孢化球蟲卵囊。自免疫接種後每日觀察鳥隻死亡情形及檢查每克糞中卵囊之排放量,結果各低劑量免疫組及對照組均沒有任何鳥隻死亡。而於免疫接種後第18天,以高劑量120,000個芽孢化卵囊攻擊,並逐日觀察記錄免疫接種後10日內之死亡情形及每克糞中卵囊之排放量,結果經低劑量免疫接種後各免疫組存活率均在92.86%以上,而未經低劑量免疫接種之陽性對照組存活率僅有10.71%;而且經低劑量免疫接種之各組,在高劑量攻擊後每克糞中所排放之卵囊量,僅為原低劑量免疫接種排放量之10.22~19.30%。由此可見,免疫接種低劑量芽孢化球蟲卵囊以作為環頸雉球蟲免疫控制之可行性是極高的。

文摘要(Summary)
From July 1997 to March 1999, coccidial infection in Galliforme birds in a total of 19 zoos and pheasant farms in Taiwan were investigated by fecal examination. A total of 395 fecal samples from 41 species of birds were collected. From flotation method examinations, 96 fecal samples (24.3%) were found to contain coccidial oocysts. After sporulation, the oocysts from the 96 fecal samples examined showed 75 samples belonging to the genus Eimeria (78.1%), 24 samples to the genus Isospora (25.0%), 8 samples to the genus Gousseffia (8.3%) and 5 samples to the genus Pythonella (5.2%). This is the first report of the genera Gousseffia and Pythonella in Taiwan.
The non-sporulated oocysts of Eimeria colchici, a cecal coccidium of pheasants, were obtained by artificial inoculation; these oocysts were divided equally and treated with following 13 kinds of chemicals for one hour. After the treatment, the oocysts were washed by distilled water and centrifuged for 4 times to eliminate the remaining chemicals. They were then incubated at 28℃ constantly for 36 hours to calculate the sporulation rates of each test group and compared them to the rates of control group. The results indicated that treating with ammonia solution showed the best inhibition effect on oocysts sporulation (92.85%), the second was phenol (38.22%) and the third was hyamine (18.73%). After the sporulation, the E. colchici oocysts, which were obtained by the same way as the above, were treated with the same 13 kinds of chemicals for one hour, washed and centrifuged for 5 times and artificial inoculated to 14 days old pheasants, the orally inoculation dose was 120,000 oocysts. Compared to the control group, the mortality of Ammonia solution treated group was lowest, being 16.67% (3/18), and in other groups (including positive control) all the inoculated pheasants died(18/18).
Unsporulated oocysts of E. colchici, the cecal coccidium of Phasianus colchicus, were collected from the faeces after artificial inoculation. These and other groups of unsporulated oocysts from the cecal mucosa were immersed in ammonia solutions in different concentration (0.75%, 0.25%, 0.125% and 0.025%) for 1 hour, then washed with distilled water and centrifuged twice for 4 times to eliminate the remained ammonia. After incubating at a constant temperature of 28℃ for 36 hours, the sporulation ratios between ammonia-treated groups and negative control had been calculated. The results indicated that the stronger the concentration of ammonia solution, the better the inhibition effect to the sporulation of oocyst. Unsporulated oocysts collected from faeces were immersed in 4 different concentration of ammonia solutions, and the sporulation-inhibition ratio from high concentration to low one are 99.82%, 37.14%, 28.55% and 0% in order. The unsporulated oocysts collected from cecal mucosa were treated as the above, and the results are 99.68%, 96.42%, 57.81% and 26.05% respectively. It shows that if the concentration of ammonia solution is below 0.25%, the unsporulated oocysts which had been excreted from animals have more resistance to it than the ones which stay in the cecal mucosa still; besides, the ammonia solution in concentration lower than 0.025% will not have the capacity to kill the oocysts which have been excreted from animals.
Non-sporulated oocysts of E. colchici, when exposed to ozone showed lower sporulation ratio than the untreated one. Lower sporulation ratio was observed, with longer ozone exposure time. Among the 14-day-old Phasianus colchicus that were each orally inoculated with 120,000 sporulated oocysts which have been exposed to ozone for 15, 30 and 60 minutes, lower mortality were observed for those inoculated with oocysts treated with ozone for longer, period of time and the oocysts in feces per gram was also less than the untreated control group. Thus ozone treatment of oocyst had been shown to inhibit the growth and infectivity of E. colchici.
Low-dose immunization of pheasants with live E. colchici oocysts was observed to be effective against massive challenge infection. Fourteen-day-old pheasants, in groups of 28, were orally inoculated with about 100, 250, 500, 1,000 and 2,000 sporulated oocysts. These birds were then challenged homologously with about 120,000 sporulated oocysts 18 days after the immunization. During the 12 days observation period after the challenge infection, over 92.86% of the immunized pheasants were still alive, but only 10.71% of the non-immunized control birds survived the challenge infection. Moreover, the number of oocysts produced after challenge infection of the immunized gro

封面
目次
表次
圖次
中文摘要
英文摘要
第一章 前言
第二章 文獻探討
第一節 環頸雉球蟲Eimeria colchici之生物學特性
一、環頸雉艾美球蟲之分類
二、Eimeria colchici之生活史
三、Eimeria colchici之致病性
四、Eimeria colchici之宿主專一性
第二節 鶉雞類之球蟲疫情
第三節 抗球蟲藥物防治
一、在雉雞類之抗球蟲效果
二、抗球蟲藥物所遭遇之困難
第四節 理化因子對寄生蟲之控制效果
一、化學藥劑對蟲卵之殺滅效果
二、臭氧的特性及應用
三、其他物理性可殺滅寄生蟲蟲卵或減低感染力之方法
第五節 球蟲疫苗的發展以及所遭遇的困難
第三章 臺灣地區鶉雞類球蟲之寄生現況
摘要
第一節 緒言
第二節 材料與方法
一、糞材樣品採集
二、球蟲卵囊檢查
第三節 結果
第四節 討論
第四章 常用化學性消毒劑對環頸雉球蟲Eimeria colchici 卵囊之殺滅效力
摘要
第一節 緒言
第二節 材料與方法
一、不同化學藥劑對Eimeria colchici卵囊芽孢化之抑制試驗
(一)環頸雉來源與飼養管理
(二)球蟲株增殖
(三)未芽孢化卵囊之製備
(四)未芽孢化卵囊藥物浸漬處理
(五)卵囊芽孢化比率計算
(六)統計分析
二、不同化學藥劑對已芽孢化Eimeria colchici卵囊之殺滅試驗
(一)已芽孢化卵囊之藥物浸漬處理
(二)人工感染試驗
(三)臨床記錄
第三節 結果
第四節 討論
第五章 氨水對環頸雉球蟲Eimeria colchici卵囊之殺滅效力
摘要
第一節 緒言
第二節 材料與方法
一、環頸雉來源與飼養管理
二、未芽孢化卵囊之製備
三、氨水浸漬處理
四、卵囊芽孢化比率計算..
五、統計分析
第三節 結果
第四節 討論
第六章 臭氧處理對環頸雉球蟲Eimeria colchici 卵囊之殺滅效力
摘要
第一節 緒言
第二節 材料與方法
一、臭氧處理對Eimeria colchici 卵囊芽孢化之抑制性試驗
(一)環頸雉來源與飼養管理
(二)球蟲株增殖
(三)未芽孢化卵囊臭氧曝氣處理
(四)臭氧殘留濃度測定
(五)卵囊芽孢化比率計算
(六)統計分析
二、已芽孢化Eimeria colchici 卵囊經臭氧曝氣處理後對宿主之病害性試驗
(一)已芽孢化卵囊之臭氧曝氣處理
(二)人工感染試驗
(三)臨床記錄與統計分析
第三節 結果
第四節 討論
第七章 免疫接種低劑量Eimeria colchici活毒卵囊以評估環頸雉球蟲病免疫控制之可行性研究
摘要
第一節 緒言
第二節 材料與方法
一、環頸雉來源與飼養管理
二、球蟲株增殖
三、低劑量人工免疫接種
四、高劑量攻擊感染
五、臨床記錄
六、統計分析
第三節 結果
第四節 討論
第八章 結論與建議
參考文獻
附錄一、球蟲寄生現況調查受檢鶉雞類鳥禽中文名對照名錄

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