發(fā)布時間：2018-03-12 18:33

  本文選題：犬真菌性皮膚病　切入點：動物模型　出處：《南京農業(yè)大學》2015年碩士論文　論文類型：學位論文

【摘要】：犬真菌性皮膚病是由真菌黏附于犬表皮及附屬結構引起的,真菌在附著之后對角質蛋白組織寄生或腐生從而迅速繁殖,最終導致局部皮膚出現脫毛、鱗屑、糜爛等病變。淺層真菌是犬真菌皮膚病的主要病原體,基本為皮膚癬菌,可分為毛癬菌屬(如須癬毛癬菌)、表皮癬菌屬(如絮狀表皮癬菌)和小孢子菌屬(如犬小孢子菌、石膏樣孢子菌)三個屬,馬拉色菌等少數酵母菌也可以導致發(fā)病。該病在各類型犬只和各個地區(qū)均常發(fā)生,多為條件致病性疾病,種類多樣,治療期較長,治愈率不高。近年來其發(fā)病率的增加可能和寵物臨床抗生素及免疫抑制劑使用量過大有關。人類也可能被犬傳染上該疾病,在抵抗力低下的特殊人群里該病發(fā)生率正在上升。對犬真菌性皮膚病的相關研究是動物醫(yī)學界目前的科研熱點,但是相關工作往往直接使用犬作為實驗動物,具有價格高、操作復雜、對操作人員造成人身威脅等缺點,本試驗嘗試建立實驗動物模型模擬犬真菌性皮膚病病變以求改善相關缺點。相關研究多集中于該病的診斷和防治,很少涉及致病機理方面的研究,本試驗嘗試探索在實驗動物機體內免疫系統與病原的相互作用,或能對其致病機理有初步的了解。試驗一:須毛癬菌皮膚病實驗動物模型的建立為了制作安全、方便、可靠的真菌性皮膚病實驗動物模型以替代犬投入相關試驗,采用在動物背部接種須毛癬菌菌懸液的方法建立了小鼠和豚鼠兩種動物的須毛癬菌皮膚病模型,每間隔1d觀察局部皮膚病變癥狀并評分,在接種真菌后第3d、第13 d、第26 d制作局部皮膚石蠟病理切片。為了了解機體免疫狀態(tài)對真菌致病的影響,在接種前分別使用地塞米松和黃芪多糖作為免疫抑制劑與免疫加強劑注射動物進行對比。結果顯示,全部豚鼠及86.7%的地塞米松組小鼠須毛癬菌再培養(yǎng)呈現陽性,建模成功。須毛癬菌對實驗動物皮膚的侵害作用在接種真菌后13 d-17 d逐漸達到最高峰,并在之后逐漸減輕癥狀。豚鼠的病變癥狀明顯而直觀,癥狀較重,而小鼠的病變癥狀較淺。黃芪多糖組的感染豚鼠對比免疫抑制組豚鼠皮膚病變癥狀評分減少。該試驗初步證明須毛癬菌實驗動物模型是可行的,動物機體的免疫狀態(tài)對須毛癬菌感染造成影響,感染后半個月左右為病變最嚴重時期。試驗二:體外藥敏試驗和療效試驗為了確定須毛癬菌皮膚病豚鼠模型應用于抗真菌藥物療效學試驗的可行性,先以棋盤微量稀釋方法進行了須毛癬菌對兩性霉素B、特比萘芬、伊曲康唑以及后兩種藥物聯合用藥的體外藥敏試驗,以肉眼進行評判,再在豚鼠模型上用藥進行藥效學評估,結果顯示聯合用藥的MIC值最低,須毛癬菌消除率最高,癥狀總積分減少最多,初步判定藥物療效在動物試驗和體外藥敏試驗中基本一致,該模型可用于相關研究。試驗三:相對熒光定量PCR法檢測細胞因子為了 了解動物機體免疫系統在真菌感染后的變化情況,探究真菌皮膚病致病機理,采取須毛癬菌皮膚病模型豚鼠第3d、第13d、第26d的血液,用相對熒光定量PCR方法檢測血液總IL-10、IL-1、TNF-α、IFN-γ四種細胞因子的含量,發(fā)現Th2細胞因子IL-10在感染須毛癬菌后第3d升高,提示動物機體在剛受到感染時處于免疫抑制狀態(tài),而Th1細胞因子IL-1、TNF-α、IFN-γ分別在第3d、第13 d和第26d達到數值高峰,提示細胞免疫系統迅速作出積極應對抵抗真菌入侵,令病情趨于好轉。
[Abstract]:Fungal skin diseases caused by fungal adhesion to canine epidermis and accessory structure, when attached to fungal keratin tissue saprophytic to multiply rapidly, resulting in local skin hair removal, scaling, erosion and other diseases. The shallow fungi were the main pathogens of canine fungal skin diseases, as the basic dermatophyte. Can be divided into the genus Trichophyton (such as Trichophyton mentagrophytes and Epidermophyton) (such as Epidermophyton floccosum) and microsporon (such as Microsporum gypseum, spores) three genus Malassezia yeasts and a few can also cause disease. The disease often occurs in all various types of dogs and various regions, for the conditions of pathogenic disease, species diversity, longer treatment, the cure rate is not high. In recent years the incidence rate increased and pet clinical antibiotic and immune inhibitor use is too large. Humans may also be infected with the disease in dogs In particular, people in low resistance to the disease incidence rate is rising. The related research on fungal skin disease is the current research hotspot of animal medicine, but the work is often used directly in dogs as experimental animal, has a high price, complex operation, the disadvantages of operator personal threats, the this test attempts to establish an experimental animal model of simulated fungal skin disease in order to improve the diagnosis and treatment of defects. The researchers concentrate on the disease, rarely involves the study of pathogenic mechanism of this experiment to explore the interaction of the immune system and pathogens in animal body, or to have a preliminary understanding of the disease the mechanism. Experiment 1: Trichophyton skin disease animal model is built in order to make safe, convenient, fungal skin disease reliable experimental animal model to replace the dogs into the relevant test, recovery The Trichophyton skin disease model mice and guinea pigs of two kinds of animal is established by using the method in the animal back inoculation of Trichophyton mentagrophytes bacterial suspension, each interval of 1D observation of local skin lesions and symptoms score at 3D after inoculation of fungi, thirteenth D, twenty-sixth d to make local skin pathological to affect the immune status. Understanding of the pathogenic fungi, respectively using dexamethasone and astragalus polysaccharide as immunosuppressive agents and immune enhancing agent injection were compared in the animal before inoculation. The results showed that dexamethasone group were Trichophyton mentagrophytes all 86.7% guinea pigs and then cultured positive, successful modeling. Against the effect of Trichophyton mentagrophytes in experimental animal skin after inoculation of fungi 13 D-17 D has reached a peak, and then gradually reduce the symptoms. Disease symptoms in guinea pigs significantly and directly, severe symptoms, and disease symptoms in mice is shallow. The APS group Comparison of immune inhibition groups of guinea pigs infected guinea pig skin lesion symptom score decreased. The results showed that t.mentagrophytes experimental animal model is feasible, the immune status of the animal body's impact on Trichophyton mentagrophytes infection, infection of about half a month after the most serious lesion period. Experiment two: the test of drug sensitivity test in vitro in order to determine the feasibility and efficacy application of t.mentagrophytes skin disease in guinea pig model of antifungal efficacy test, first by the checkerboard microdilution method of terbinafine t.mentagrophytes to amphotericin B, itraconazole, drug sensitivity test in vitro and after two drug combination therapy, evaluation to the naked eye, then the medication in the guinea pig model on pharmacodynamics evaluation the results show that a combination of drugs, the lowest MIC value, t.mentagrophytes elimination rate is the highest, the total symptom score decreased most, the preliminary determination of drug efficacy in animal experiments and in vitro Drug sensitivity test is basically the same, the model can be used for related research. Experiment three: relative cytokine detection by fluorescence quantitative PCR in order to understand the animal immune system changes in fungal infection, pathogenic fungi on skin disease mechanism, take Trichophyton skin disease model of guinea pig 3D, 13D, 26D blood, with detection of IL-10, blood relative fluorescence quantitative PCR method IL-1, TNF- alpha, gamma IFN- content of four kinds of cytokines, Th2 cell factor IL-10 in 3D infection increased the Trichophyton mentagrophytes, suggesting that the animal body on immunosuppression in just the infection, while Th1 cytokines IL-1, TNF- alpha IFN-, gamma after 3D, thirteenth D and 26D to the numerical peak, the cellular immune system quickly and actively respond to fungal invasion, so the condition is getting better.

【學位授予單位】：南京農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S858.292

【參考文獻】

相關期刊論文 前10條

1 魯金波;;一例犬真菌與螨蟲混合感染并發(fā)膿皮病診治[J];中國動物保健;2013年01期

2 蒙翠萍;;一例犬皮膚真菌和螨蟲混合感染的診治報告[J];廣西畜牧獸醫(yī);2013年01期

3 朱華君;石達友;李興華;陳翱蕾;陳欣健;唐兆新;;中西藥結合治療犬貓犬小孢子菌感染皮膚病[J];黑龍江畜牧獸醫(yī);2012年24期

4 張釗偉;張坤;朱曉文;忻悅;于志君;劉玉秀;桑曉宇;程凱慧;劉紅;趙永坤;王鐵成;高玉偉;夏咸柱;;豚鼠TNF、IFN-γ、IL-1熒光定量PCR檢測方法的建立[J];中國病原生物學雜志;2012年03期

5 薛俊潔;來常海;孫淑東;周懷貴;;犬膿皮癥的診治[J];山東畜牧獸醫(yī);2012年01期

6 夏永高;劉桂清;;上海地區(qū)犬常見皮膚病的流行病學調查及防治對策[J];黑龍江八一農墾大學學報;2010年06期

7 馮秀娟;;犬真菌性皮膚病的發(fā)生與治療[J];中國工作犬業(yè);2009年12期

8 薛占永;呼秀智;林德貴;;犬膿皮病病原菌的分離鑒定[J];中國獸醫(yī)雜志;2009年04期

9 林德貴;;犬貓皮膚病[J];養(yǎng)犬;2009年01期

10 李曾夏子;李文平;;犬皮膚真菌病研究進展[J];畜牧與獸醫(yī);2009年03期

相關博士學位論文 前1條

1 趙魯杭;黃芪多糖的制備及其對巨噬細胞和樹突狀細胞免疫功能的影響[D];浙江大學;2011年

相關碩士學位論文 前5條

1 趙熠群;上海地區(qū)犬常見皮膚病的調查與治療研究[D];上海交通大學;2010年

2 屈德芳;長江三角洲地區(qū)犬貓皮膚真菌病調查及體外藥敏試驗[D];南京農業(yè)大學;2009年

3 王璐璐;真菌性角膜炎動物模型的建立及糖皮質激素對其發(fā)病過程的影響[D];鄭州大學;2007年

4 李天偉;犬皮膚真菌病和蠕形螨病的流行病學調查與臨床診療研究[D];南京農業(yè)大學;2006年

5 賀星亮;某犬場犬主要體外寄生蟲和皮膚病調查及采取的措施[D];南京農業(yè)大學;2003年

，

本文編號：1602771