广西部分家禽养殖场曲霉菌（Aspergillus sp.）遗传多态性及对伊曲康唑敏感性分析

发布时间：2021-07-22 09:35

　　曲霉属的真菌是主要存在于土壤、水以及腐败的营养物的一类霉菌。它们产生大量的孢子,这些孢子通过空气能很容易的扩散到环境中,由于这类孢子的普遍存在,人和动物不断的暴露于曲霉菌孢子中。烟曲霉和黄曲霉被认为是引起人和各类动物真菌性疾病的主要病原,相比哺乳动物而言,鸟类更易感。家禽养殖场的环境条件非常适合许多真菌,包括曲霉菌的生长于繁殖。本研究的主要日的是评估广西家禽养殖场分离的主要曲霉菌的遗传多样性及对抗真菌药物的敏感性。研究第一部分主要对南宁的3个养殖场以及桂林的一个（包含一个孵化场）种鸡场真菌污染的情况进行调查。调查主要采集家禽咽部样品及空气样本,采样时间一般持续几周,同时对孵化场进行了三个孵化周期的监测。经表型鉴定辅助分子生物学方法鉴定,最终共收集到188份烟曲霉和159份黄曲霉。第二部分建立或应用多位点数目可变串联重复序列（VNTR）多态性对黄曲霉和烟曲霉进行遗传多样性分析。本研究筛选了8个VNTR位点对黄曲霉遗传多样性进行分析,并设计双重反应。利用筛选到的8个多态性位点对91份黄曲霉分离株,其中包括6个参考株进行分型分析,结果共产生78种基因型,其中71种基因型仅出现一次,辛普森多样... 

【文章来源】：广西大学广西壮族自治区 211工程院校

【文章页数】：151 页

【学位级别】：博士

【文章目录】：
摘要
ABSTRACT
Résumé
Chapter Ⅰ Introduction and outline of the thesis
    1. The genus Aspergillus
        1.1. Ecology
        1.2. Reproduction and morphological features
        1.3. Identification of Aspergillus species
        1.4. Aspergillus fumigatus
        1.5. Aspergillus flavus
        1.6. Production of mycotoxins
    2. Molecular typing of Aspergillus species
        2.1. Random amplified polymorphic DNA(RAPD)
        2.2. Restriction fragment length polymorphism(RFLP)and related methods
        2.3. Multilocus sequence typing(MLST)
        2.4. Microsatellite length polymorphism(MLP)
        2.5. Multiple locus VNTR analysis(MLVA)
    3. Aspergillosis
        3.1. Virulence factors
        3.2. Immunity and host susceptibility
        3.3. Human aspergillosis
        3.4. Avian aspergillosis
        3.5. Treatment of fungal infections and emergence of azote resistance in Aspergillusfumigatus
    4. Importance of poultry in China and Guangxi
        4.1. Geographical context
        4.2. Chinese market of animal products
        4.3. Focus on poultry products market in China
        4.4. Poultry genetic resources in China
        4.5. Poultry systems in China
    5. Outline of the thesis
Chapter Ⅱ Fungal contamination in avian farms in Guangxi,China
    1. Introduction
    2. Material and methods
        2.1. Avian farms
        2.2. Sampling and fungal isolation
        2.3 Identification of Fungal
        2.4. Statistical analyses
    3. Results
    4. Discussion
    5. Conclusion
Chapter Ⅲ Development and use of VNTR based methods for typing Aspergillus spp
    1. Introduction
    2. Materials and methods
        2.1. Origin of fungal isolates
        2.2. DNA isolation
        2.3. Selection of VNTR markers for A.flavus
        2.4. VNTR markers for A.fumigatus
        2.5. Specificity
        2.6. Stability and reproducibility
        2.7. Discriminatory power
        2.8. Clustering analysis for A.flavus and A.fumigatus isolates
    3. Results
        3.1. Development of the MLVA method for A. flavus
        3.2. Genetic diversity of A. flavus isolates
        3.3. Genetic diversity of A. fumigatus isolates
    4. Discussion
    5. Conclusion
Chapter IV Susceptibility to itraconazole in Aspergillusisolated from avian farms in France and China
    1. Introduction
    2. Materials and methods
        2.1. Origin of fungal isolates
        2.2. Antifungal susceptibility testing
        2.3. Sequencing of Cyp21A
        2.4. MLVA typing
    3. Results
        3.1. Susceptibility to itraconazole
        3.2. Mutations of Cyp51A and corresponding protein
        3.3. MLVA typing
    4. Discussion
    5. Conclusion
General discussion and perspectives
References
Acknowledgements
致谢
Appendix
    List of abbreviations
    Glossary
Communications
Published Papers



本文编号：3296902