隱球菌（Cryptococcus species）和阿氏絲孢酵母（Tricosporon asahii）在免疫力不全病患的感染情形逐年上升。它們通常會造成肺、皮膚、腦及全身系統的感染。免疫力不全病患的罹病率和死亡率都要比正常人高。從以前的研究報告得知，新型隱球菌可從鳥類的糞便培養檢測出來。新型隱球菌grubii變異型（Cryptococcus neoformans var. grubii）（血清型A）是最主要造成臨床感染的菌株，也可在鳥類糞便中被大量培養檢測出來。新型隱球菌gattii變異型（Cryptococcus neoformans var. gattii）（血清型B、C）生存於大自然植物中。桉樹是最密切相關的樹種。其他樹種也能檢測出gattii變異型，特別是在熱帶和亞熱帶的地區。與隱球菌比較起來，阿氏絲孢酵母是土壤中的腐生黴菌。於本實驗中，我們在南台灣做了有關於隱球菌和阿氏絲孢酵母的環境調查。其中包括120個賽鴿糞便檢體和114個桉樹檢體。結果在賽鴿糞便中培養檢測出30株新型隱球菌（佔25%），4株Cryptococcus laurentii（佔3.3%），和2株Cryptococcus albidus（佔1.7%）。另外，在賽鴿糞便中培養檢測出25株阿氏絲孢酵母（佔20.8%）。但是在桉樹檢體中卻沒有檢測出任何相關菌株。經鑑定所有新型隱球菌都屬grubii變異型（血清型A）。而且這些新型隱球菌株的藥物敏感度試驗，都呈現對Amphotericin B、Fluconazole和Flucytosine敏感，而且最小抑菌濃度分別是≦0.25μg/ml、2μg/ml和≦1μg/ml。綜和而論，隱球菌和阿氏絲孢酵母在本實驗從賽鴿糞便中培養檢測出來，尤其是阿氏絲孢酵母被大量檢測出。這些菌株造成的臨床感染，是值得我們注意的，尤其是賽鴿與人接觸頻繁關係密切。未來更需要密集監測和調查，對於疾病的控制和預防上提供即時有效的資訊。

The increasing infection of Cryptococcus species and Tricosporon asahii emerged in clinical patients who were immunocompromised. They usually induce lung, skin, brain and systemic infection. Morbidity and mortality of immunocompromised patients are higher than normal healthy people. Cryptococcus neoformans var. grubii （serotype A） infections were reported in clinical cases predominantly and they were isolated from birds’ droppings in large amount. Cryptococcus neoformans var. gattii （serotype B, C） had a natural life in plants, especially Eucalypticus trees. Isolations from other trees were reported increasingly in the tropical and subtropical areas. Comparing to Cryptococcus species, Tricosporon asahii is the normal mycoses of soil. In this study, we performed an environmental investigation concerning Cryptococcus species and Tricosporon asahii in Southern Taiwan. 120 droppings of racing pigeons and 114 samples from Eucalypticus trees were obtained. The results revealed that 30 Cryptococcus neoformans were isolated from racing pigeons’ droppings （25%）, as well as 4 Cryptococcus laurentii （3.3%） and 2 Cryptococcus albidus （1.7%）. In addition, 25 Tricosporon asahii ( 20.8% ) were isolated from droppings of racing pigeons. But, none of Cryptococcus species or Tricosporon asahii is isolated from Eucalypticus trees （0%）. All of Cryptococcus neoformans isolated from pigeons’ droppings were var. grubii （serotype A） and their drug susceptibility tests showed sensitive to Amphotericin B （minimal inhibitory concentration ≦0.25μg/ml） and Fluconazole （minimal inhibitory concentration 2μg/ml） and Flucytosine （minimal inhibitory concentration ≦1μg/ml）. To sum up, both Cryptococcus species and Tricosporon asahii were isolated from droppings of racing pigeons in our study, especially Tricosporon asahii in large amount. Opportunistic infection caused by these species should be given more attention to racing pigeons which have close contact with human . Intensive investigation and surveillance should be carried out in the future to provide an information for the control and prevention of diseases.

論文審定書..........................................................................................................Ⅰ
中文摘要………………………………………………………………………Ⅱ
英文摘要……………………………………………………………………………..Ⅲ
目錄.......................................................................................................................... Ⅳ
圖目錄..................................................................................................................... Ⅵ
表目錄………………………………………………………………………………Ⅶ
緒言………………………………………………………………………………1
文獻探討……………………………………………………………………2
隱球菌流行病學……………………………………………………………………2
隱球菌分離和鑑定…..……………………………………………………………..3
隱球菌臨床症狀…………………………………………………………..……......4
隱球菌臨床治療……………………………………………………………..……..5
阿氏絲孢酵母流行病學………………………………………………………..…..5
阿氏絲孢酵母分離和鑑定……………………………………………………..…..6
阿氏絲孢酵母臨床症狀……………………………………………………..……..7
阿氏絲孢酵母臨床治療……………………………………………………………7
實驗材料和方法............................................................................................................8
實驗材料…………………………………………………………..…..………………8
檢體來源………………………………………………………………...……...…..8
新型隱球菌標準菌株…………………………………………………………..…..8
菌株培養分離和鑑定系統…………………………………………………..……..8
菌株DNA萃取和聚合酶連鎖反應（PCR）和電泳的試劑..................................9
實驗方法……………………………………………………..………………………..9
菌株的分離……………………………………………………………......………..9
菌株的鑑定……………………………………………………………………..…10
菌株的藥敏試驗……………………………………………………………..……10
菌株的保存…………………………………………………………………..……10
菌株DNA的萃取………………………………………………………………....10
聚合酶連鎖反應（PCR）與電泳…………………………………………….…..11
結果………………………………………………………………………..…………12
鳥糞檢體培養檢測分離結果……………………………………………..………12
桉樹檢體培養檢測分離結果…………………………………………………..…12
新型隱球菌和阿氏絲孢酵母DNA萃取分離………………………………..…..12
新型隱球菌DNA指紋雜合試驗（PCR-fingerprinting）和血清分型………….. ..13
新型隱球菌藥物敏感試驗……………………………………………………..…13
阿氏絲孢酵母聚合酶連鎖反應（PCR）……………….………………….…....13
討論….……………………………………………………………………………….14
參考文獻.………………………………………………………………………….....19
附錄…….………………………………………………………………………….....38

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