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請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/43650
完整後設資料紀錄
DC 欄位值語言
dc.contributor.advisor龐飛(Victor Fei Pang)
dc.contributor.authorChung-Ling Liuen
dc.contributor.author劉家伶zh_TW
dc.date.accessioned2021-06-15T02:25:09Z-
dc.date.available2010-08-19
dc.date.copyright2009-08-19
dc.date.issued2009
dc.date.submitted2009-08-18
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/43650-
dc.description.abstract全球性兩棲動物族群數量呈現明顯下降趨勢,蛙壺菌(Batrachochytrium dendrobatidis)感染可能是主要元兇之一,此病原主要感染有尾目及無尾目,目前發現有14科250種以上兩棲動物感染蛙壺菌症,帶原者有蝌蚪、美國牛蛙 (Rana catesbeiana)、非洲爪蟾 (Xenopus laevis)、蔗蟾 (Bufo marinus) 及虎斑鈍口螈 (Ambystoma tigrinum)。受到蛙壺菌感染而帶有病原,但卻無明顯的症狀,並可能將病原傳給其他兩棲動物,這些動物稱之為帶原者 (carrier)。本研究之目的為以組織病理學及PCR診斷技術針對回溯性及持續收集之病例進行檢測,以了解蛙壺菌症是否入侵台灣及其發生的情形。回溯性以組織病理學檢查1993到2007年,台北市立動物園送檢的64隻蛙類及36隻蠑螈,發現於2003年12月21日送檢的迷彩箭毒蛙 (Dendrobates auratus) 為第1例檢出有蛙壺菌感染症之病例。現況調查分為三大類,(ㄧ)外來種寵物蛙:總數278隻,檢體包括新鮮皮膚檢體14個、皮膚棉花拭子109個、蛙類飼養箱的水樣132個及石蠟包埋組織塊99個,經由PCR檢測及組織病理學診斷,共有17個陽性病例,其中含4隻角蛙 (Ceratophrys sp.)、4隻南美角蛙 (Ceratophrys crawelli)、3隻黃金角蛙 (Ceratophrys cranwelli, alphosis)、2隻鐘角蛙 (Ceratophrys ornate)、2隻猴樹蛙 (Phyllomedusa hypochondrialis) 及2隻白化非洲爪蟾 (Xenopus laveis),陽性病例主要以角花蟾屬的蛙類為主,佔總陽性病例76.4% (13/17);分別以各種蛙類的樣本總數估算陽性率,角花蟾蛙類的平均檢出陽性率為12.9% (13/101), 白化非洲爪蟾 (Xenopus laveis ) 為5.7% (2/35),猴樹蛙 (Phyllomedusa hypochondrialis) 為33.3% (2/6)。(二)食用蛙:總數113隻,檢體包括新鮮皮膚35個、皮膚棉花拭子36個、蛙類飼養池的水樣36個及石蠟包埋組織蠟塊50個,PCR及組織病理學檢測結果皆為陰性。(三)野外蛙隻:總數為443隻,檢體包括石蠟包埋組織蠟塊393個、新鮮皮膚檢體9個、皮膚棉花拭子39個、蝌蚪口器棉花拭子2個及蛙類飼養池的水樣32個;PCR檢測及組織病理學檢測結果皆為陰性。研究結果顯示,蛙壺菌症已藉由外來種寵物蛙的輸入而入侵台灣,惟目前尚無證據顯示本病已散播至食用蛙及野外蛙隻。zh_TW
dc.description.abstractBatrachochytrium dendrobatidis is the cause of chytridiomycosis, a potentially fatal epidermal infection in amphibian. The fungus infects 2 amphibian orders (Anura and Caudata), 14 families, and at least 250 species. Bull frog (Rana catesbeiana), Xenopus laevis, tadpoles, tiger salamander (Ambystoma tigrinum), and cane toad (Bufo marinus) are known carriers. To determine whether chytridiomycosis has intruded Taiwan, a survey was carried to examine the infecion by histopathology and/or polymerase chain reaction (PCR) in the imported pet frogs, salamanders, meat frogs and field frogs. Retrospectively, a total of 64 frogs and 36 salamanders dying spontaneously and necropsied routinely at Taipei Zoo during the years of 1993 and 2007 were subjected to evaluation. During this period, only one Dendrobates auratus died in December of 2003, which was diagnosed as a mild infection of chytrid fungus histopathologically. For imported pet frogs, a total of 278 frogs were examined by light microscopy, hostopathology and /or PCR, specimens including 14 fresh skin samples, 109 skin swabs, 132 bath water samples and 99 paraffin-embedded skin tissues. Seventeen positive cases were diagnosed, including 4 Ceratophrys sp., 4 Ceratophrys crawelli, 3 Ceratophrys cranwelli, alphosis, 2 Ceratophrys ornate, 2 Phyllomedusa hypochondrialis and 2 Xenopus laveis. Ceratophrys sp. seemed to be the major species affected by chytridiomycosis with a positive rate of 76.4% (13/17), and the overall positive rates of Ceratophrysl, Xenopus laveis, Phyllomedusa hypochondrialis were 12.9% (13/101), 5.7% (2/35), and 33.3% (2/6), respectively. For meat frogs, a total of 35 fresh skin samples, 35 skin swabs, 36 bath water samples, and 50 paraffin-embedded tissues were collected from 113 frogs. For field frogs, a total of 9 fresh skin sampels, 39 skin swabs, 2 mouthpart swab, 32 bath water samples, and 393 paraffin-embedded tissues were collected from 443 frogs. However, all specimens from meat frogs and field frogs were negative by light microscopy, histopathology and/or PCR. The results of the investigation indicate that chytridiomycosis has indeed been introduced into Taiwan possibly via the importation of pet frogs. Utile now, no evidence of spreading chytridiomycosis to meat frogs and field frogs has been revealed.en
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dc.description.tableofcontents摘要 I
Abstract II
圖次 VII
表格 XI
第一章 序言 1
第一節 蛙壺菌 2
1.1 兩棲動物消失危機與新興傳染性疾病 2
1.2 蛙壺菌分類 4
1.3 蛙壺菌特性及生活史 4
1.4 蛙壺菌培養、物理及化學特性 5
1.4.1 液態培養基種類及配方 5
1.4.2 固態培養基種類及配方 6
1.4.3 培養方式 6
1.4.4 菌株生長及保存方式 7
1.4.5 致死溫度 7
1.4.6 培養特性 7
第二節 蛙壺菌症 8
2.1 背景 8
2.2 全球蛙壺菌的感染狀況 9
2.2.1 非洲地區 9
2.2.2 美洲地區 9
2.2.3 歐洲地區 10
2.2.4 澳洲地區 10
2.2.5 亞洲地區 10
2.3 蛙壺菌症 11
2.4 蛙壺菌傳播方式12
2.5 致病機轉 14
2.6 潛在協同因子 16
2.7 實驗室診斷方法 17
2.7.1 實驗室感染診斷機率 17
2.7.2 病理學檢查 18
2.7.3 免疫化學染色 18
2.7.3.1 直接接合抗體法(direct conjugate-labeled antibody method)18
2.7.3.2 鹼性磷酸-抗鹼性磷酸酶複合物法(簡稱APAAP 法)配合
Holland’s trichrome keratin 染色 19
2.7.4 掃描式電子顯微鏡 20
2.7.5 穿透式電子顯微鏡 20
2.8 疾病監控及防範措施 20
2.8.1 國際防範措施 20
2.8.2 殺滅蛙壺菌 21
2.8.3 消毒劑使用 21
2.9 治療方式 22
2.10 兩棲類常見其他皮膚病及其致病原之類症鑑別 23
Table 2.1 Characteristics of various fungal dermatitis in amphibian 27
第三節 台灣進口資料 28
第四節 台灣寵物店販售的外來兩棲類之概況 28
第五節 兩棲動物保育行動計劃 29
第三章 材料與方法 34
第一節 實驗設計及流程圖 34
第二節 實驗材料 36
2.1 回溯性研究 36
2.2 現況調查之檢體來源 36
2.3 現況調查之檢體收集方式 37
第三節 實驗方法 39
3.1 DNA 之萃取 39
3.1.1 皮膚棉花棒拭子及蝌蚪口器棉花拭子之真菌核酸萃取 39
3.1.2 蛙類飼養箱或養殖池的水質樣本之真菌核酸萃取 40
3.1.3 福馬林固定、石蠟包埋組織之真菌核酸萃取 41
3.1.4 冷凍皮膚檢體及冷凍蝌蚪口器之真菌核酸萃取 42
3.1.5 組織病理學檢查 43
3.1.6 組織化學染色 43
3.1.7 穿透式電子顯微鏡 44
3.1.8 掃描式電子顯微鏡 44
第四節 聚合酶鏈鎖反應(PCR)45
4.1 Batrachochytrium dendrobatidis 之聚合酶鏈鎖反應溶液成分及條件 45
4.2 Batrachochytrium dendrobatidis 之聚合酶鏈鎖反應產物電泳 45
4.3 聚合酶鏈鎖反應產物的直接核酸定序 46
第四章 結果 46
第一節 回溯性調查 46
1.1 台北市立動物園剖檢兩棲動物之樣本背景 46
1.2 病理學檢查 47
1.3 石蠟組織包埋蠟塊之PCR 檢測結果 48
1.4 動物園陽性病例之發生狀況 48
第二節 現況兩棲動物樣本 49
第三節 食用蛙類調查 49
3.1 食用蛙類的樣本背景 49
3.2 組織切片病理診斷結果 49
3.3 分子生物檢測結果 50
第四節 野外蛙類調查 50
4.1 野外蛙類樣本之背景 50
4.2 組織病理切片診斷結果 51
4.3 分子生物檢測結果 51
第五節 外來種寵物蛙類調查 51
5.1 外來種寵物蛙類樣本之背景 51
5.2 外來種寵物蛙類感染蛙壺菌症之臨床症狀 52
5.3 皮膚抹片檢查 53
5.4 病理學檢查 53
5.5 聚合酶鏈鎖反應結果 54
5.6 其他實驗室診斷項目結果 54
5.6.1 掃描式電子顯微鏡檢查 (Scanning electron microscopy) 54
5.6.2 穿透式電子顯微鏡檢查(Transmission electron microscopy) 55
第六節 國內食用蛙、野外蛙類及外來種寵物蛙的調查結果 55
第七節 陽性病例之背景資料及統計分析 56
7.1 台北市立動物園陽性病例之背景資料 56
7.2 B 家寵物店發生背景資料 57
7.3 蛙壺菌病例統計分析 58
第五章 討論 59
第六章 參考文獻 69
Table 4-1 List of anura necropsied in Taipei zoo 77
Appendix 1. The results of histopathological examination and PCR of skin from various species of urodela and anura dying spontaneously and subjected to necropsy in Taipei Zoo during 1995 to 2007. 1
Aappendix 2. The results of histopathological examination and PCR of skin from various species of urodela and anura dying spontaneously and subjected to necropsy from field frogs, imported pet frogs and meat frogs during January, 2008 to April, 2009. 5
dc.language.isozh-TW
dc.subject無尾目zh_TW
dc.subject兩棲動物zh_TW
dc.subject蛙類zh_TW
dc.subject蛙壺菌zh_TW
dc.subject蛙壺菌症zh_TW
dc.subjectchytridiomycosisen
dc.subjectanuraen
dc.subjectfrogen
dc.subjectamphibianen
dc.subjectBatrachochytrium dendrobatidisen
dc.subjectchytriden
dc.title台灣外來種及本土種兩棲動物感染蛙壺菌之概況zh_TW
dc.titleChytrid fungus (Batrachochytrium dendrobatidis) infection in field and imported amphibian in Taiwanen
dc.typeThesis
dc.date.schoolyear97-2
dc.description.degree碩士
dc.contributor.coadvisor鄭謙仁(Chian-Ren Jeng)
dc.contributor.oralexamcommittee蕭世烜(Shih-Hsuan Hsiao),劉涓(Jiuan Liu),張志成(Chang, Chih-Cheng)
dc.subject.keyword兩棲動物,蛙類,蛙壺菌,蛙壺菌症,無尾目,zh_TW
dc.subject.keywordBatrachochytrium dendrobatidis,chytrid,chytridiomycosis,amphibian,frog,anura,en
dc.relation.page97
dc.rights.note有償授權
dc.date.accepted2009-08-18
dc.contributor.author-college獸醫專業學院zh_TW
dc.contributor.author-dept獸醫學研究所zh_TW
顯示於系所單位:獸醫學系

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