不同猪鼻支原体菌株的检测分离和比较以及实验感染模型的初步研究
发布时间：2021-06-15 15:46
尽管支原体缺乏细胞壁,而且只有很小的基因组,但各属的支原体都能够引起不同宿舍的慢性感染。猪鼻支原体的较高感染性可能是由于猪的生长环境发生了变化,而更大程度上因为猪肉生产系统向专业化,集约化演变而导致。然而,猪鼻支原体是通过鼻子对鼻子来传播,通过感染猪的接触或谷仓、农场中的空气传播等。虽然M.猪鼻是猪的上呼吸道中正常菌群的一部分,但它具有致病性。它是关节炎,胸膜炎,心包炎和腹膜炎的病原体,并容易引起乳猪的地方性肺炎。大多数感染在亚临床和临床症状表现上可能有所不同,而且混合感染也很常见。菌株毒力的变化,天气的变化,还有病原体和宿主的免疫系统等这些机制都是系统性的。在基因组水平,有报道称不同牛群中分离出来的具有极大的异质性。在一个农场中的同一动物中分离出来的不同菌株,可能在毒力或引起干扰和交叉反应方面有所不同。在新陈代谢中,猪鼻支原体基因中的一些变化可以逃避宿主的免疫系统,这可能有助于其感染。目前的研究主要集中在从不同方面来控制猪鼻支原体。（1）猪病原体的检测主要是猪鼻支原体和猪肺炎支原体。临床样品为7-35天的猪鼻拭子,主要是来自中国的江苏省,他们从不同患病牛群中收集了病原体,利用巢式PC...
【文章来源】：南京农业大学江苏省 211工程院校 教育部直属院校
【文章页数】：119 页
【学位级别】：硕士
【文章目录】：
摘要
ABSTRACT
CHAPTER 1 LITERATURE REVIEW
1.1. TAXONOMY AND A GENERAL INTRODUCTION TO MYCOPLASMAS
1.2. ECOLOGY AND PATHOGENESIS OF MYCOPLASMA
1.2.1 Evasion of the Immune system
1.2.2 Interaction with the Host Cells and Cytopathology
1.2.3 M.hyorhinis Major Immunogen,P37
1.3. ETIOLOGYY
1.4. EPIDEMIOLOGY
1.5. CLINICAL PRESENTATION OF MHR INFECTION IN SWINE
1.6. DIAGNOSTIC TECHNIQUES FOR THE DETECTION OF MHR
1.6.1 Diagnosis
1.6.2 Macroscopic and microscopic examination
1.6.3 Fluorescent antibody and immunohistochemistry assays
1.6.4 Culture
1.6.5 Serology
1.6.6 Polymerase chain reaction(PCR)
1.7. BIOSECURITY AND AN INTEGRATED APPROACH FOR THE MANAGEMENT OF MHR
1.8. THE SWINE INDUSTRY IN KENYAAND CHINA
1.9. PROBLEM STATEMENT
1.10 RATIONALE
1.11 HYPOTHESIS
1.12 OBJECTIVES OF THE STUDY
CHAPTER 2 DETECTION OF RESPIRATORY PATHOGENS M.hyorhinis AND M.hyopneumoniae BY NESTEDPCR IN PORCINE NASAL SWABS FROM CLINICALLY INFECTED HERDS
2.1 INTRODUCTION
2.2 MATERIALS AND METHODS
2.2.1 Animals
2.2.2 Sample preparation
2.2.3 Nested PCR
2.2.4 Statistical analysis
2.3 RESULTS
2.4 DISCUSSION
CHAPTER 3 DETECTION,ISOLATION AND IDENTIFICATION OF M.hyorhinis AND ASSESSING THEANTIMICROBIAL PROFILES OF THE ISOLATES
3.1 DETECTION,ISOLATION AND IDENTIFICATION OF MHR
3.1.1 Introduction
3.1.2 Materials and Methods
3.1.3 Results
3.2 ISOLATION OF MHR STRAINS
3.2.1 Introduction
3.2.2 Materials and Methods
3.2.3 Results
3.2.4 Discussion
3.3. THE IN VITRO ASSESSMENT OF THE ANTIMICROBIAL PROFILES OF 8 MHR ISOLATES AGAINST
3.3.1 Introduction
3.3.2 Materials and Methods
3.3.3 Results
3.3.4 Discussion
CHAPTER 4 IMMUNOGENICITY TESTING IN MICE AND DETERMINATION OF ANTIBODY TITERS BYINDIRECT-ELISA
4.1 INTRODUCTION
4.1.1 Freund's adjuvant
4.1.2 Immunoglobulins
4.2 MATERIALS AND METHODS
4.2.1 Antigen Preparation
4.2.2 Quantification of Antigen by CCU
4.2.3 Quantification of Antigen by BCA
4.2.4 Quantification of Antigen using qPCR
4.2.5 Results
4.2.6 Immunization
4.2.7 Detection of serum IgG antibody
4.2.7.1 Preparation of coating protein
4.2.7.2 Indirect IgG ELISA Assay(Tween-20 ELISA)
4.2.8 Statistical Analysis
4.3 RESULTS
4.4 DISCUSSION
CHAPTER 5 PRIMARY STUDY OF INFECTION AND DEVELOPMENT OF A CHALLENGE MODEL FOR M.hyorhinis-INDUCED PATHOGENESIS IN SWINE
5.1 INTRODUCTION
5.2 MATERIALS AND METHODS
5.2.1 Animals
5.2.2 Experiment 1
5.2.3 Experiment 2
5.2.4 Experiment 3
5.2.5 Sample collection and Detection
5.2.6 Histopathological examination
5.3 RESULTS
5.3.1 Experiment 1
5.3.2 Experiment 2
5.3.3 Experiment 3
5.4 DISCUSSION
CHAPTER 6 VIRULENCE SCREENING OF M. hyorhinis STRAINS THROUGH MORTALITY INDEX OFEXPERIMENTALLY INFECTED CHICKEN EMBRYOS
6.1 INTRODUCTION
6.2 MATERIALS AND METHODS
6.2.1 Quantification of the strains by CCU
6.2.2 Inoculation
6.2.3 Detection by Culture,PCR and Histopathology
6.2.4 Statistical analysis
6.3 RESULTS
6.3.1 Mortality
6.3.2 Clinical Presentation
6.3.3 Detection by Culture and PCR
6.3.4 Histopathology
6.4 DISCUSSION
REFERENCES
OVERALL CONCLUSION
INNOVATION
LIST OF PUBLICATIONS
ACKNOWLEDGEMENTS
ABBREVIATIONS
APPENDIX
【参考文献】：
期刊论文
[1]猪鼻支原体vlp重复区融合蛋白的构建表达及反应原性分析[J]. 熊祺琰,纪燕,刘占军,马庆红,冯志新,刘茂军,白方方,邵国青. 中国人兽共患病学报. 2013(09)
[2]Identification of antigens by monoclonal antibody PD4 and its expression in Escherichia coli[J]. Jin-Ying Ning Guo-Xun Sun Su Huang Hong Ma Ping An Lin Meng Shu-Mei Song Jian Wu Cheng-Chao Shou, Department of Biochemistry and Molecular Biology, School of Oncology and Beijing Institute for Cancer Research, Peking University, Beijing 100034, China. World Journal of Gastroenterology. 2003(10)
本文编号：3231337
【文章来源】：南京农业大学江苏省 211工程院校 教育部直属院校
【文章页数】：119 页
【学位级别】：硕士
【文章目录】：
摘要
ABSTRACT
CHAPTER 1 LITERATURE REVIEW
1.1. TAXONOMY AND A GENERAL INTRODUCTION TO MYCOPLASMAS
1.2. ECOLOGY AND PATHOGENESIS OF MYCOPLASMA
1.2.1 Evasion of the Immune system
1.2.2 Interaction with the Host Cells and Cytopathology
1.2.3 M.hyorhinis Major Immunogen,P37
1.3. ETIOLOGYY
1.4. EPIDEMIOLOGY
1.5. CLINICAL PRESENTATION OF MHR INFECTION IN SWINE
1.6. DIAGNOSTIC TECHNIQUES FOR THE DETECTION OF MHR
1.6.1 Diagnosis
1.6.2 Macroscopic and microscopic examination
1.6.3 Fluorescent antibody and immunohistochemistry assays
1.6.4 Culture
1.6.5 Serology
1.6.6 Polymerase chain reaction(PCR)
1.7. BIOSECURITY AND AN INTEGRATED APPROACH FOR THE MANAGEMENT OF MHR
1.8. THE SWINE INDUSTRY IN KENYAAND CHINA
1.9. PROBLEM STATEMENT
1.10 RATIONALE
1.11 HYPOTHESIS
1.12 OBJECTIVES OF THE STUDY
CHAPTER 2 DETECTION OF RESPIRATORY PATHOGENS M.hyorhinis AND M.hyopneumoniae BY NESTEDPCR IN PORCINE NASAL SWABS FROM CLINICALLY INFECTED HERDS
2.1 INTRODUCTION
2.2 MATERIALS AND METHODS
2.2.1 Animals
2.2.2 Sample preparation
2.2.3 Nested PCR
2.2.4 Statistical analysis
2.3 RESULTS
2.4 DISCUSSION
CHAPTER 3 DETECTION,ISOLATION AND IDENTIFICATION OF M.hyorhinis AND ASSESSING THEANTIMICROBIAL PROFILES OF THE ISOLATES
3.1 DETECTION,ISOLATION AND IDENTIFICATION OF MHR
3.1.1 Introduction
3.1.2 Materials and Methods
3.1.3 Results
3.2 ISOLATION OF MHR STRAINS
3.2.1 Introduction
3.2.2 Materials and Methods
3.2.3 Results
3.2.4 Discussion
3.3. THE IN VITRO ASSESSMENT OF THE ANTIMICROBIAL PROFILES OF 8 MHR ISOLATES AGAINST
3.3.1 Introduction
3.3.2 Materials and Methods
3.3.3 Results
3.3.4 Discussion
CHAPTER 4 IMMUNOGENICITY TESTING IN MICE AND DETERMINATION OF ANTIBODY TITERS BYINDIRECT-ELISA
4.1 INTRODUCTION
4.1.1 Freund's adjuvant
4.1.2 Immunoglobulins
4.2 MATERIALS AND METHODS
4.2.1 Antigen Preparation
4.2.2 Quantification of Antigen by CCU
4.2.3 Quantification of Antigen by BCA
4.2.4 Quantification of Antigen using qPCR
4.2.5 Results
4.2.6 Immunization
4.2.7 Detection of serum IgG antibody
4.2.7.1 Preparation of coating protein
4.2.7.2 Indirect IgG ELISA Assay(Tween-20 ELISA)
4.2.8 Statistical Analysis
4.3 RESULTS
4.4 DISCUSSION
CHAPTER 5 PRIMARY STUDY OF INFECTION AND DEVELOPMENT OF A CHALLENGE MODEL FOR M.hyorhinis-INDUCED PATHOGENESIS IN SWINE
5.1 INTRODUCTION
5.2 MATERIALS AND METHODS
5.2.1 Animals
5.2.2 Experiment 1
5.2.3 Experiment 2
5.2.4 Experiment 3
5.2.5 Sample collection and Detection
5.2.6 Histopathological examination
5.3 RESULTS
5.3.1 Experiment 1
5.3.2 Experiment 2
5.3.3 Experiment 3
5.4 DISCUSSION
CHAPTER 6 VIRULENCE SCREENING OF M. hyorhinis STRAINS THROUGH MORTALITY INDEX OFEXPERIMENTALLY INFECTED CHICKEN EMBRYOS
6.1 INTRODUCTION
6.2 MATERIALS AND METHODS
6.2.1 Quantification of the strains by CCU
6.2.2 Inoculation
6.2.3 Detection by Culture,PCR and Histopathology
6.2.4 Statistical analysis
6.3 RESULTS
6.3.1 Mortality
6.3.2 Clinical Presentation
6.3.3 Detection by Culture and PCR
6.3.4 Histopathology
6.4 DISCUSSION
REFERENCES
OVERALL CONCLUSION
INNOVATION
LIST OF PUBLICATIONS
ACKNOWLEDGEMENTS
ABBREVIATIONS
APPENDIX
【参考文献】：
期刊论文
[1]猪鼻支原体vlp重复区融合蛋白的构建表达及反应原性分析[J]. 熊祺琰,纪燕,刘占军,马庆红,冯志新,刘茂军,白方方,邵国青. 中国人兽共患病学报. 2013(09)
[2]Identification of antigens by monoclonal antibody PD4 and its expression in Escherichia coli[J]. Jin-Ying Ning Guo-Xun Sun Su Huang Hong Ma Ping An Lin Meng Shu-Mei Song Jian Wu Cheng-Chao Shou, Department of Biochemistry and Molecular Biology, School of Oncology and Beijing Institute for Cancer Research, Peking University, Beijing 100034, China. World Journal of Gastroenterology. 2003(10)
本文编号：3231337
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