声明
Contents
List of figures
List of tables
Abbreviations
摘要
Abstract
Chapter 1 Chicken Coccidiosis and Immune Response to Eimeria
1．1．Introduction
1．2．Economic importance
1．3．The life cycle of an Eimeria parasite
1．4．Diagnosis
1．4．1．Clinical findings
1．4．2．Gross lesions
1．5．1．Species
1．5．2．Extent of contamination
1．6．Chicken coccidial organelles antigen-related functions
1．7．Immune response to coccidiosis
1．7．2．Immune response mechanisms in avian coccidiosis
1．8．Objectives of the study
References
Chapter 2 Control of Chicken Coccidiosis
2．1．Management control
2．2．Chemoprophylaxis
2．3．Feed quality control
2．4．Vaccination
2．4．1．Biology of vaccination with special reference to Eimeria
2．4．2．Immune responses to vaccination Eimeria infection
2．4．3．Vaccines available against coccidiosis
References
Chapter 3 Isolation and Identification of Eimeria brunetti strain
Abstract
3．1．Introduction
3．2．Materials and methods
3．2．1．Parasites and chickens
3．2．2．Equipments,solutions,chemicals,reagents and buffers
3．2．3．Isolation of oocyst
3．2．4．Identification of Eimeria species by PCR amplification
3．3．Results
3．4．Discussion
References
Chapter 4 Cloning，Expression and Characterization of Eimeria brunetti Microneme-2 Protein(EbMIC2)
Abstract
4．1．Introduction
4．2．Materials and methods
4．2．1．Vectors and bacteria strains
4．2．3．Designing primers
4．2．4．Animals and parasites
4．2．5．Chicken immune sera against E． brunetti
4．2．7 Reverse transcription and amplification of cDNA (RT-PCR)
4．2．8．Cloning of EbMIC2 gene
4．2．9．Sequence alignments and phylogenetic analysis
4．2．10．Construction of recombinant expression vector of EbMIC2
4．2．1 1．Expression and purification of recombinant EbMIC2
4．2．12．Immuno-blot for the recombinant EbMIC2 and native protein of EbMIC2
4．3．Results
4．3．1．Cloning of E. brunetti MIC2 gene
4．3．2．Percent identity,phylogenetic tree and antigenic analysis
4．3．3．Expression and purification of recombinant EbMIC2 protein
4．3．4．Immunoblot for the recombinant EbMIC2 and native protein of EbMIC2
4．4．Discussion
References
Chapter 5 Immunogenieity of Eimeria brunetti Microneme-2 Protein and Its Protective Effects on Chicken
Abstract
5．1．Introduction
5．2．Materials and methods
5．2．5．Detection of the expressions of EbMIC2 gene in vivo by reverse transcription polymerase chain reaction (RT-PCR) assay and western blot analysis
5．2．6．Determination of serum antibody level by enzyme-linked Immuno Sorbent assay (ELISA)
5．2．8．Production of anti-serum for passive immunization
5．2．9．Immunization and challenge experiment
5．2．10．Immunization and challenge experiment in passive immunization
5．2．11．Evaluation of immune protection
5．2．12 Statistical analysis
5．3．Results
5．3．2．Detection of the transcription of constructed recombinant plasmid pVAX1-EbMIC2 in vivo
5．3．4．IgG and cytokine levels in sera of immunized chickens
5．3．5．Protective effects of EbMIC2 against E．brunetti challenge in active immunization
5．4．Discussion
References
Chapter 6 Cloning，Expression and Characterization of Eimeria brunetti Apical Membrane Antigen-1(EbAMA1)
Abstract
6．1．Introduction
6．2．Materials and Methods
6．3．Results
6．3．1．Cloning of E．brunettiAMAl gene
6．3．2．Sequence analysis
6．3．3．Percent identity,phylogenetic tree and antigenic analysis
6．3．4．Construction and identification of the recombinant pET-28a(+)-EbAMA1
6．3．5．Expression and purification of recombinant EbAMA1 protein
6．3．6 Immunoblot for the recombinant EbAMA1 and native protein of EbAMA1
6．4．Discussion
References
Chapter 7 Immunogenicity of Eimeria brunetti Apical Membrane Antigen-1 and Its Protective Effects on Chicken
Abstract
7．1．Introduction
7．2．Materials and Methods
7．3．Results
7．3．2．Detection of the transcription of constructed recombinant plasmid EbAMA1 in vivo
7．3．4．IgG and cytokine levels in sera of immunized chickens
7．3．5．Protective effects of EbAMA1 against E．brunetti challenge in active immunization
7．4．Discussion
References
Conclusion
Acknowledgements
