Molecular Epidemiology of Piroplasms in Pakistan, and Molecu
发布时间：2021-03-26 04:44
泰勒虫和巴贝斯虫是蜱传性红细胞内原生寄生虫,可感染世界范围内大多数家畜。这些寄生虫主要感染热带和亚热带地区的牛羊,导致畜牧业巨大的经济损失。虽然,在巴基斯坦已经确定一些梨形虫种类感染奶牛和水牛,然而,多数尚未在分子水平上确认。考虑到这些寄生虫的重要性,本文运用反向线性杂交试验（RLB）在巴基斯坦Punjab的Sheikhupura和Okara地区对奶牛和水牛进行分子流行病学调查,评价泰勒虫和巴贝斯虫种类的流行性和传播动态。实验对具有遗传多样性的18S rRNA基因和ITS序列进行了研究。简要地说,来自采样区的102份黄牛和水牛血液样本分别保存于血纸中,运输到中国武汉华中农业大学,农业微生物学国家重点实验室进行DNA的提取；并进行18S rRNA的V4高变区扩增和RLB分析。61份（59.80%）样品用巴贝斯虫／泰勒虫属特异性探针进行杂交试验,61份样品中仅检测到环形泰勒虫。六份样品只与泰勒虫／巴贝斯虫特异性探针杂交,而不与任何其他种属特异性探针杂交,暗示有新的物种或基因型存在。对这6份样品进行18srRNA和ITS序列的扩增,6份样品的18S rRNA序列和ITS序列与环形泰勒虫最接近...
【文章来源】：华中农业大学湖北省 211工程院校 教育部直属院校
【文章页数】：130 页
【学位级别】：博士
【文章目录】：
Contents
Abbreviations
Abstract
Chapter 1-Review of Literature
1.1 Babesia and Theileria
1.1.1 General Introduction
1.1.2 Life cycle
1.1.3 Pathophysiology and clinical manifestation
1.1.4 Epidemiology
1.1.4.1 Geographical distribution
1.1.4.2 Transmission dynamics
1.1.5 Advances in diagnosis
1.1.5.1 Various types of PCR
1.1.5.2 Loop mediated isothermal amplification assay
1.1.5.3 Reverse line blot hybridization assay
1.1.5.4 Role of molecular biology in the detection of new species
1.1.6 Conclusions and hypothesis
1.2 Heat Shock Protein 90（HSP90）
1.2.1 General Introduction
1.2.2 Structure of HSP90
1.2.2.1 N-terminal domain
1.2.2.2 Middle domain
1.2.2.3 C-terminal domain
1.2.3 Function of HSP90
1.2.4 HSP90 in apicomplexan parasites
1.2.5 Conclusions and hypothesis
Chapter 2-Molecular Epidemiology of Theileria annulata in Pakistan
2.1 Introduction
2.2 Materials and methods
2.2.1 Study area and sample collection
2.2.2 Epidemiological survey
2.2.3 DNA extraction
2.2.4 Reverse Line Blot Hybridization assay
2.2.4.1 Polymerase chain reaction
2.2.4.2 Preparation of membrane
2.2.4.3 Reverse line blot hybridization
2.2.5 Cloning and sequencing of 18S rRNA gene and ITS regions
2.2.6 Phylogenetic and sequence analysis
2.2.7 Statistical analysis
2.3 Results
2.3.1 Prevalence and RLB results
2.3.2 Nucleotide sequence heterogeneity analysis
2.3.3 Phylogenetic analysis
2.3.4 Associated risk factors
2.4 Discussion
Chapter 3-Two Putative HSP90 Proteins in Babesia orientalis
3.1 Introduction
3.2 Materials and methods
3.2.1 In silico identification of two HSP90-like proteins of B. orientalis
3.2.2 Parasites and animals
3.2.3 DNA and RNA extraction and preparation of cDNA
3.2.4 Amplification,cloning and sequencing of BoHSP90-A and BoHSP90-B genes
3.2.5 Sequence structure,function and motifs prediction
3.2.6 Recombinant expression and purification of BoHSP90-A and BoHSP90-B
3.2.7 Production of anti-rBoHSP90-A and anti-rBoHSP90-B immune serum
3.2.8 SDS-PAGE and western blotting
3.3 Results
3.3.1 Molecular characterization and sequence analysis
3.3.2 Predicted structure and functional analysis
3.3.3 HSP90 inhibitors
3.3.4 Antigenicity analysis
3.3.5 Identification of BoHSP90-A and BoHSP90-B
3.4 Discussion
Chapter 4-Conclusions and Future Prospects
References
List of Tables
List of Figures
APPENDICES
ACKNOWLEDGEMENT
PUBLICATIONS
【参考文献】：
期刊论文
[1]Detection of Babesia bovis in blood samples and its effect on the hematological and serum biochemical profile in large ruminants from Southern Punjab[J]. Samreen Zulfiqar,Sadia Shahnawaz,Muhammad Ali,Arif Mahmood Bhutta,Shahid Iqbal,Sikandar Hayat,Shazia Qadir,Muhammad Latif,Nazia Kiran,Ali Saeed,Muhammad Ali,Furhan Iqbal. Asian Pacific Journal of Tropical Biomedicine. 2012(02)
[2]First molecular detection of Theileria ovis in Rhipicephalus sanguineus tick in Iran[J]. Telmadarraiy Zakkyeh,Oshaghi Mohammad Ali,Hosseini Vasoukolaei Nasibeh,Yaghoobi Ershadi Mohammad Reza,Babamahmoudi Farhang,Mohtarami Fatemeh. Asian Pacific Journal of Tropical Medicine. 2012(01)
[3]寄生于中国水牛Bubalus Vbubalis的巴贝斯虫一新种（梨形虫目:巴贝斯科）[J]. 刘钟灵,赵俊龙,马丽华,姚宝安. 畜牧兽医学报. 1997(01)
本文编号：3100974
【文章来源】：华中农业大学湖北省 211工程院校 教育部直属院校
【文章页数】：130 页
【学位级别】：博士
【文章目录】：
Contents
Abbreviations
Abstract
Chapter 1-Review of Literature
1.1 Babesia and Theileria
1.1.1 General Introduction
1.1.2 Life cycle
1.1.3 Pathophysiology and clinical manifestation
1.1.4 Epidemiology
1.1.4.1 Geographical distribution
1.1.4.2 Transmission dynamics
1.1.5 Advances in diagnosis
1.1.5.1 Various types of PCR
1.1.5.2 Loop mediated isothermal amplification assay
1.1.5.3 Reverse line blot hybridization assay
1.1.5.4 Role of molecular biology in the detection of new species
1.1.6 Conclusions and hypothesis
1.2 Heat Shock Protein 90（HSP90）
1.2.1 General Introduction
1.2.2 Structure of HSP90
1.2.2.1 N-terminal domain
1.2.2.2 Middle domain
1.2.2.3 C-terminal domain
1.2.3 Function of HSP90
1.2.4 HSP90 in apicomplexan parasites
1.2.5 Conclusions and hypothesis
Chapter 2-Molecular Epidemiology of Theileria annulata in Pakistan
2.1 Introduction
2.2 Materials and methods
2.2.1 Study area and sample collection
2.2.2 Epidemiological survey
2.2.3 DNA extraction
2.2.4 Reverse Line Blot Hybridization assay
2.2.4.1 Polymerase chain reaction
2.2.4.2 Preparation of membrane
2.2.4.3 Reverse line blot hybridization
2.2.5 Cloning and sequencing of 18S rRNA gene and ITS regions
2.2.6 Phylogenetic and sequence analysis
2.2.7 Statistical analysis
2.3 Results
2.3.1 Prevalence and RLB results
2.3.2 Nucleotide sequence heterogeneity analysis
2.3.3 Phylogenetic analysis
2.3.4 Associated risk factors
2.4 Discussion
Chapter 3-Two Putative HSP90 Proteins in Babesia orientalis
3.1 Introduction
3.2 Materials and methods
3.2.1 In silico identification of two HSP90-like proteins of B. orientalis
3.2.2 Parasites and animals
3.2.3 DNA and RNA extraction and preparation of cDNA
3.2.4 Amplification,cloning and sequencing of BoHSP90-A and BoHSP90-B genes
3.2.5 Sequence structure,function and motifs prediction
3.2.6 Recombinant expression and purification of BoHSP90-A and BoHSP90-B
3.2.7 Production of anti-rBoHSP90-A and anti-rBoHSP90-B immune serum
3.2.8 SDS-PAGE and western blotting
3.3 Results
3.3.1 Molecular characterization and sequence analysis
3.3.2 Predicted structure and functional analysis
3.3.3 HSP90 inhibitors
3.3.4 Antigenicity analysis
3.3.5 Identification of BoHSP90-A and BoHSP90-B
3.4 Discussion
Chapter 4-Conclusions and Future Prospects
References
List of Tables
List of Figures
APPENDICES
ACKNOWLEDGEMENT
PUBLICATIONS
【参考文献】：
期刊论文
[1]Detection of Babesia bovis in blood samples and its effect on the hematological and serum biochemical profile in large ruminants from Southern Punjab[J]. Samreen Zulfiqar,Sadia Shahnawaz,Muhammad Ali,Arif Mahmood Bhutta,Shahid Iqbal,Sikandar Hayat,Shazia Qadir,Muhammad Latif,Nazia Kiran,Ali Saeed,Muhammad Ali,Furhan Iqbal. Asian Pacific Journal of Tropical Biomedicine. 2012(02)
[2]First molecular detection of Theileria ovis in Rhipicephalus sanguineus tick in Iran[J]. Telmadarraiy Zakkyeh,Oshaghi Mohammad Ali,Hosseini Vasoukolaei Nasibeh,Yaghoobi Ershadi Mohammad Reza,Babamahmoudi Farhang,Mohtarami Fatemeh. Asian Pacific Journal of Tropical Medicine. 2012(01)
[3]寄生于中国水牛Bubalus Vbubalis的巴贝斯虫一新种（梨形虫目:巴贝斯科）[J]. 刘钟灵,赵俊龙,马丽华,姚宝安. 畜牧兽医学报. 1997(01)
本文编号：3100974
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