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研究生:汪鴻展
研究生(外文):Hung-Chan Wang
論文名稱:西高地白梗犬上皮發育不良症繼發上皮感染
論文名稱(外文):Epidermal Dysplasia of West Highland White Terrier with Secondary Epidermal Infections
指導教授:張清棟張清棟引用關係
指導教授(外文):Ching-Dong Chang
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:2005
畢業學年度:93
語文別:中文
論文頁數:121
中文關鍵詞:西高地白梗犬上皮發育不良穿透式電子顯微鏡
外文關鍵詞:West Highland White TerrierEpidermal dysplasiaTEM
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西高地白梗犬上皮發育不良症為一特殊之皮膚性疾病,於1989年Scott及Miller首次提出報告,並指出罹病犬隻一般於6月齡時開始呈現臨床症狀,症狀初期以皮膚潮紅、搔癢及脫毛為主,並引起自我創傷性炎症反應及繼發性皮膚感染症且對藥物治療成效反應不佳,相關於本病之研究迄今仍相當有限,台灣迄今尚無本病相關研究之報告。本研究針對兩頭罹患上皮發育不良之西高地白梗犬進行研究,此兩頭患犬均為雌性且具同胎血緣關係,且發病年齡均為其9~10月齡時,皮膚病灶初期呈現搔癢及脫毛,臨床治療一年中,對抗生素及類固醇等藥物反應不良,即使投予抗黴菌藥物及合併使用含ketoconazole洗毛劑時,症狀亦無改善,皮膚增厚形成多處皺摺,且惡化成為慢性嚴重性皮膚病;血液學檢查無明顯異常。由此兩頭患犬體表各部位進行病原微生物分離培養,並以傳統微生物生化鑑定方法及聚合酶鏈鎖反應技術進行病原鑑定,結果顯示兩頭患犬均繼發上皮Staphylococcus intermedius及Malassezia pachydermatis感染。由皮膚組織病理學方式觀察皮膚生檢樣本,發現異常之發育不良皮膚上皮細胞,其細胞大小、形狀及染色性均與正常表皮細胞不同,此外,由Staphylococcus intermedius及Malassezia pachydermatis繼發感染之病變亦可被觀察。本研究亦首次藉由穿透式電子顯微鏡觀察西高地白梗犬上皮發育不良細胞之超顯微構造,結果發現,主要為基底細胞與棘細胞發生型態學上變異,多型性畸型之變異細胞明顯有別於正常細胞結構外,棘細胞核仁內之染色質分散於核膜四周且核膜皺摺而凹入核質,形成裂隙樣結構;而基底細胞正常應具豐富之胞器,但發育不良之細胞其胞質內所含之胞器不一,胞質稀少,明顯有所差異。本病是否為西高地白梗犬此品系之遺傳性疾病仍需進一步探討,希望藉由本研究中所建立之相關資料,提供日後針對本病之後續研究參考用,期待未來更進一步明瞭本病之致病機轉。
Epidermal Dysplasia of West Highland White Terrier is a special skin disorder that starts in rather young dogs, from a few weeks to a few months of age, and often is found on six months. Early clinical sings were skin erythema, pruritus, alopecia, self perpetuating inflammation and secondary Staphylococcus intermedius and Malassezia pachydermatis infections. There are limited articles about Epidermal Dysplasia of West Highland White Terrier and there is no report of Epidermal Dysplasia of West Highland White Terrier in Taiwan so far. Two, female, litter-mate West Highland White Terrier dogs were studied. Early clinical sings were skin erythema, pruritus, and variable alopecia. These dogs were treated with antibiotics, steroids and ketoconazole shampoo for one year and clinical signs didn’t improve in these dogs. Staphylococcus intermedius and Malassezia pachydermatis were isolated from epidermis of both dogs and were molecularly confirmed by PCR and DNA sequences. Dermatohistopathologically, epidermal dysplasia, anisocytosis, cell pleomorphism, various cell staining characteristics and cell atypia of Epidermal Dysplasia of West Highland White Terrier with secondary Staphylococcus intermedius and Malassezia pachydermatis infections were also observed. Results of transmission electron microscope examination showed that cell atypia were observed in the basal and prickle cells. We still don’t know if Epidermal Dysplasia of West Highland White Terrier is genetic-related disease. The present study data may be used for further investigation for the exact mechanism of Dysplasia of West Highland White Terrier.
中文摘要...............................................I
Abstract............................................III
誌謝...................................................V
目錄..................................................VI
圖表目錄...............................................X
第1章 前言.............................................1
第2章 文獻回顧..........................................2
2.1 發育不良.........................................2
2.1.1 西高地白梗犬之上皮發育不良症世界首報.............2
2.2 瑞士之病例報告....................................7
2.3 葡萄球菌之介紹....................................9
2.3.1 葡萄球菌之特性................................9
2.3.2 葡萄球菌之菌落形態及顯微鏡下形態...............10
2.3.3 葡萄球菌之生化反應及鑑定方式...................10
2.3.4 犬隻葡萄球菌性膿皮症..........................10
2.3.5 葡萄球菌性膿皮症之治療........................11
2.3.6 葡萄球菌性感染之診斷原則......................12
2.4 Malassezia pachydermatis之介紹.................13
2.4.1 M. pachydermatis之概述.....................13
2.4.2 分類與命名...................................14
2.4.3 形態學.......................................14
2.4.4 超顯微構造....................................15
2.4.5 培養生理特性..................................15
2.4.6 流行病學......................................17
2.4.6.1 M. pachydermatis在皮膚疾病所扮演之角色.....17
2.4.6.2 M. pachydermatis與犬外耳炎及皮膚炎之關係...18
2.4.6.3 人類之M. pachydermatis感染................18


第3章 材料與方法.........................................20
3.1 病例之蒐集........................................20
3.1.1 病歷之問取....................................20
3.1.2 病例之常規理學檢查及血液學檢查...................20
3.2 培養基之製備.......................................20
3.2.1 Blood agar...................................20
3.2.2 MacConkey agar...............................21
3.2.3 Sabouraud dextrose agar ( SDA )..............21
3.2.4 Modified Dixon’s agar ( MDA )...............21
3.3 微生物分離、純化....................................22
3.4 生化鑑定...........................................22
3.4.1 葡萄球菌於API® Staph System之生化學鑑定..........22
3.4.2 抗菌劑紙錠瓊脂擴散試驗...........................23
3.4.3 Malassezia pachydermatis之鑑定................23
3.5 分子生物學檢查DNA之萃取..............................24
3.5.1 葡萄球菌DNA萃取.................................24
3.5.2 Malassezia pachydermatis DNA萃取..............24
3.6 引子對之選擇與設計...................................25
3.7 聚合酶鏈鎖反應 ( Polymerase Chain Reation;PCR ).....25
3.7.1 Staphylococcus intermedius PCR敏感性試驗.......26
3.7.2 Staphylococcus intermedius PCR特異性試驗.......26
3.8 PCR產物之確認......................................27
3.8.1 PCR產物之純化與定序.............................27
3.9 菌種序列之比對......................................27
3.10 皮膚生檢...........................................28
3.10.1 肉眼病理學檢查..................................28
3.10.2 組織病理學檢查..................................28
3.11 特殊染色...........................................29
3.11.1 Periodic Acid Schiff ( PAS )染色..............29
3.11.2 Brown and Brenn stain革蘭氏染色...............29
3.12 生檢皮膚組織病理學病變紀錄...........................30
3.13 電子顯微鏡檢查......................................31
3.13.1 穿透式電子顯微鏡 ( Transmission Electron Microscope;
TEM ) 檢查.....................................31
3.13.2 掃描式電子顯微鏡 ( Scanning Electron Microscope;
SEM ) 檢查.....................................31
3.13.2.1皮膚生檢SEM樣本之製備..............................31
3.13.2.2 M. pachydermatis培養菌落SEM樣本之製備.............32
第4章 結果................................................33
4.1 病歷之蒐集及問取.....................................33
4.2 病例之常規理學檢查及血液學檢查.........................33
4.2.1 病例1之理學檢查結果.............................33
4.2.2 病例1之血液學檢查結果...........................34
4.2.3 病例2之理學檢查結果.............................34
4.2.4 病例2之血液學檢查結果...........................34
4.3 微生物培養與鑑定結果.................................34
4.3.1 Blood agar及MacConkey agar培養結果.............34
4.3.2 API® Staph System生化鑑定結果..................43
4.3.3 抗菌劑紙錠瓊脂擴散試驗結果.......................43
4.3.4 Malassezia pachydermatis培養結果...............43
4.4 分子生物學檢查.......................................50
4.4.1 Staphylococcus intermedius之聚合酶鏈鎖反應結果...50
4.4.2 Staphylococcus intermedius之聚合酶鏈鎖反應敏感性結果..51
4.4.3 Staphylococcus intermedius之聚合酶鏈鎖反應特異性結果..51
4.4.4 Staphylococcus intermedius PCR產物之定序........53
4.4.5 Malassezia pachydermatis之聚合酶鏈鎖反應結果.....56
4.4.6 Malasseiza pachydermatis PCR產物之定序..........56
4.5 皮膚生檢之組織病理學檢查結果...........................61
4.5.1 病例1皮膚生檢結果................................61
4.5.2 病例2皮膚生檢結果................................74
4.5.3 特殊染色結果.....................................81
4.6 電子顯微鏡檢查結果.....................................83
4.6.1 皮膚樣本穿透式電子顯微鏡檢查結果....................83
4.6.2 皮膚樣本掃描式電子顯微鏡檢查結果....................90
4.6.3 M. pachydermatis培養大小菌落之掃描式電子顯微鏡
檢查結果..........................................94
第5章 討論..................................................97
參考文獻....................................................103
附錄.......................................................109
作者簡介....................................................121
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