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研究生:賴政宏
研究生(外文):Cheng-Hung Lai
論文名稱:台灣中部地區犬心絲蟲症之研究:流行病學及致病機轉
論文名稱(外文):Study of dirofilariasis in central Taiwan: epidemiology and pathogenesis
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:90
語文別:中文
論文頁數:135
中文關鍵詞:犬心絲蟲流浪犬白線斑蚊熱帶家蚊胞外基質基質金屬酵素
外文關鍵詞:Dirofilaria immitisstray dogsAedes albopictusCulex quinquefasciatusextracellular matrixmatrix metalloproteinases
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犬心絲蟲(Dirofilaria immitis)是犬隻血液循環體系內最重要之寄生蟲之一,其成蟲寄生於犬隻肺動脈及右心室,對犬隻健康會有嚴重的影響。過去台灣地區犬心絲蟲的寄生感染相關研究均以北部地區為主,依其結果顯示近年來犬心絲蟲症在北部地區迅速的蔓延,流浪犬間的感染率更是居高不下。然而台灣中部地區之相關研究並不多,因此本研究特別針對台灣中部地區的流浪犬及病媒蚊進行犬心絲蟲的感染狀況調查,並探討自然媒介蚊種之感受性及微絲蟲濃度對蚊子之影響。在所收集的218隻流浪犬(雄性115隻、雌性103隻)中,經由血液抹片檢查呈微絲蟲陽性者72隻,其陽性率為33.03%,而經由心臟剖檢後發現132隻流浪犬心臟有心絲蟲成蟲存在,感染率為60.55%。雄犬感染率為62.61%(72/115)、雌犬感染率為58.25%(60/103)。雌、雄犬隻感染率並無明顯差異,但是有27.52%流浪犬為潛伏性感染。患犬10 ml血液內之微絲蟲數為0至635隻,心臟內之成蟲數為1至49隻不等,其中有22隻(16.67%)流浪犬為單性感染。此結果顯示台灣中部地區亦已成為犬心絲蟲症之流行區域,因此在人畜共通公共衛生學上之潛在危險性頗值得密切注意。
在以誘蚊燈誘捕收集之五種蚊子中僅發現熱帶家蚊(Culex quinquefasciatus)和白線斑蚊(Aedes albopictus)會被犬心絲蟲感染,其感染率分別為4.28%和3.74%,而三斑家蚊(Cx. tritaeniorhynchus)、環紋家蚊(Cx. annulus)和白腹叢蚊(Armigeres subalbatus)均未被感染。平均每隻被感染之白線斑蚊體內可發現3.43隻犬心絲蟲幼蟲,高於熱帶家蚊的2.89隻。在餵食每公撮含有7500隻微絲蟲之犬血後,熱帶家蚊攝入之微絲蟲數量約為白線班蚊的二倍(平均值分別為31.73和16.47)。然而白線斑蚊體內第三期幼蟲之發育數量幾乎是熱帶家蚊的三倍(平均值為3.25和1.10),其病媒效用指數(vector efficiency index, VEI)分別為19.73和3.47。這些結果顯示熱帶家蚊和白線斑蚊是台灣中部地區犬心絲蟲之自然病媒。雖然白線斑蚊在犬心絲蟲之傳播上較有效率,然而考慮其族群數量及攝食習性認為熱帶家蚊可能在台灣中部犬心絲蟲症的流行上扮演較重要的角色。將白線斑蚊和熱帶家蚊分別餵食含有不同微絲蟲濃度的犬血(2500至25000 mf/mL)後觀察其發育及對蚊體之影響。犬心絲蟲幼蟲於兩種蚊種體內之發育並無明顯不同。雖然熱帶家蚊攝入較多微絲蟲,然而白線斑蚊體內發育之幼蟲數則明顯高於熱帶家蚊。吸飽血的白線斑蚊之死亡率明顯高於熱帶家蚊,而且較高微絲蟲濃度會提高二者之死亡率。白線斑蚊在各種微絲蟲濃度下之VEI均高於熱帶家蚊,然而其存活時間卻明顯縮短。因此,推測有低微絲蟲濃度之犬心絲蟲患犬是造成病媒蚊感染之主要來源。
犬心絲蟲之感染寄生會造成犬隻心臟肥大擴張、心內膜肥厚,形態上可見心臟圓形化、心室壁變薄、心肌細胞產生退行性變化和溶解現象。心肌組織的主要結構包括心肌細胞以及細胞間的胞外基質(extracellular matrix, ECM),而心臟之胞外基質主要由第一型和第三型膠原蛋白(collagen)所組成。心肌細胞間之膠原蛋白一直被認為在維持心臟之結構與功能上扮演著重要的角色,因此膠原蛋白組成的改變可能會影響心臟之功能。將犬心絲蟲患犬之心肌組織切片經過浸式鍍銀染色後觀察,可發現心肌間質之膠原蛋白的含量減少並呈斷離現象。胞外基質金屬酵素(matrix metalloproteinases, MMPs或稱為matrixins)可以分解胞外基質之蛋白成分。出現在心肌的MMPs可以降解心臟細胞外基質的組成,在心臟重建的過程中扮演重要角色。取心肌組織進行受質凝膠電泳(gelatin-substrate zymography)後發現犬心絲蟲患犬左、右心室之72kDa(MMP-2)和92kDa(MMP-9)明膠溶解帶之強度較正常者明顯增加,而且以右心室之含量較多。這兩種酵素均為鈣離子(Ca2+)依賴性,其最適合之酸鹼值為6至9,而且其活性均會被1, 10-phenanthroline完全抑制。這些結果顯示犬心絲蟲的感染會導致心肌細胞基質金屬酵素之大量表現,造成心肌間質組織中膠原蛋白的重建進而改變心臟之結構與功能。因此抑制心肌細胞MMPs的表現或許可以在犬心絲蟲感染過程中控制心肌組織的重建、維持心肌之正常功能。
Dirofilaria immitis is one of the most important parasites of canine, which resides primarily in the pulmonary arteries and right ventricle and may severely affect canine health. Researches about dirofilariasis in Taiwan has been conducted in the northern area and demonstrated that the incidence of dirofilariasis is progressively increasing. However, no pertinent information was available in central Taiwan. This study first investigates the prevalence of dirofilariasis in stray dogs and mosquitoes from central Taiwan, the susceptibility of naturally infected mosquitoes to D. immitis and the effect of microfilarial density on the viability of mosquitoes. A total of 218 stray dogs (115 males and 103 females) were collected from central Taiwan and examined. The numbers of positive results from blood smear and autopsies were 72 and 132, respectively. The overall prevalence was 60.55%, 62.61% in males and 58.25% in females. The number of microfilariae in 10 mL blood and adult worms in the hearts of infected dogs ranged from 0 to 635 and from 1 to 49, respectively. These results indicated that the central Taiwan has became endemic area of dirofilariasis.
In five species of mosquitoes collected by using light traps, only Culex quinquefasciatus and Aedes albopictus were infected with D. immitis; the prevalence was 4.28% and 3.74%, respectively, while Cx. tritaeniorhyn- chus, Cx. annulus and Armigeres subalbatus were not infected with D. immitis. The intensity of D. immitis found in Ae. albopictus (3.43 larvae/mosquito) surpassed that found in Cx. quinquefasciatus (2.89 larvae/mosquito). After being fed with canine blood containing 7500 microfilariae (mf) per ml, Cx. quinquefasciatus ingested approximately twice as many mf as Ae. albopictus (mean of 31.73 in comparison to 16.47). However, almost triple the number of third-stage infective larvae developed in Ae. albopictus compared to Cx. quinquefasciatus (mean of 3.25 as compared with 1.10), with a vector efficiency index (VEI) of 19.73 and 3.47, respectively.
Ae. albopictus and Cx. quinquefasciatus were fed canine blood with different microfilarial density of D. immitis ranging from 2500 to 25,000 microfilariae/mL. Larval development in these two mosquito species did not differ significantly. Although Cx. quinquefasciatus ingested more microfilariae, the number of larvae which developed in Ae. albopictus invariably exceeded that in Cx. quinquefasciatus. Mortality of the engorged Ae. albopictus was significantly greater than that of Cx. quinquefasciatus, and higher microfilarial density elevated the mortality in both species. Although the vector efficiency index of Ae. albopictus exceeded that of Cx. quinquefasciatus at all microfilarial densities, its survival time was markedly reduced. These results indicated that Cx. quinquefasciatus and Ae. albopictus serves as natural vectors of D. immitis in central Taiwan. Although Ae. albopictus was more efficient for heartworm transmission, the investigation found after considering the vector population and feeding habits that Cx. quinquefasciatus may play a more prominent role in transmitting dirofilariasis in central Taiwan. Finally, dogs with low microfilarial density are implicated as the main source for transmitting D. immitis from dogs to mosquitoes.
Heartworm infection may cause dilatation of the heart and hypertrophy of the endocardium. Morphologically, the heart became round and the thickness of the ventricle decreased. Microscopic examination revealed cardiocytic degeneration and verrucous endocarditis. The heart consists of cardiocytes and the interstitial extracellular matrix (ECM). The ECM consists mainly of type I and type III collagens in the form of fibrils that preserve tissue architecture and chamber geometry. The present study revealed that the ECM content of heart were reduced in heartworm-infected dogs and became fragmented and dissociated by employing the silver impregnation stain. Normal and heartworm infected dogs displayed a considerable difference in quantity of collagen. The matrix metalloproteinases (MMPs), also called matrixins, are considered to play a central role in the breakdown of ECM. An obvious escalation of gelatinolytic activity of 72 (MMP-2) and 92 kDa (MMP-9) were noted in both right and left ventricle in the gelatin-substrate zymography of heartworm-infected dogs, and the enzymatic intensity of the right ventricle exceeded that of the left ventricle. These enzymes were Ca2+-dependent, the optimal pH for these enzymes ranged from 6 to 9, and the presence of 1, 10-phenanthroline completely inhibited their activity. These results indicate that heartworm infection could up-regulate the expression of MMP-2 and MMP-9, and then result in the remodeling of ECM and alter the architecture and function of the heart. Thus, regulation of myocardial MMPs expression may be an important therapeutic target to control myocardial matrix remodeling in the setting of dirofilariasis.
目 錄
表次 II
圖次 IV
中文摘要 1
英文摘要 5
第一章 前言 9
第二章 文獻探討 13
第三章台灣中部地區流浪犬心絲蟲感染狀況調查 56
第四章台灣中部地區自然感染蚊種對犬心絲蟲之感受性 66
第五章微絲蟲濃度對犬心絲蟲病媒蚊之影響 78
第六章基質金屬酵素與犬心絲蟲致病機制之相關性 87
第七章結論 106
參考文獻 109
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