發(fā)布時間：2021-03-26 04:44

　　泰勒蟲和巴貝斯蟲是蜱傳性紅細(xì)胞內(nèi)原生寄生蟲,可感染世界范圍內(nèi)大多數(shù)家畜。這些寄生蟲主要感染熱帶和亞熱帶地區(qū)的牛羊,導(dǎo)致畜牧業(yè)巨大的經(jīng)濟(jì)損失。雖然,在巴基斯坦已經(jīng)確定一些梨形蟲種類感染奶牛和水牛,然而,多數(shù)尚未在分子水平上確認(rèn)。考慮到這些寄生蟲的重要性,本文運(yùn)用反向線性雜交試驗(yàn)（RLB）在巴基斯坦Punjab的Sheikhupura和Okara地區(qū)對奶牛和水牛進(jìn)行分子流行病學(xué)調(diào)查,評價泰勒蟲和巴貝斯蟲種類的流行性和傳播動態(tài)。實(shí)驗(yàn)對具有遺傳多樣性的18S rRNA基因和ITS序列進(jìn)行了研究。簡要地說,來自采樣區(qū)的102份黃牛和水牛血液樣本分別保存于血紙中,運(yùn)輸?shù)街袊錆h華中農(nóng)業(yè)大學(xué),農(nóng)業(yè)微生物學(xué)國家重點(diǎn)實(shí)驗(yàn)室進(jìn)行DNA的提��；并進(jìn)行18S rRNA的V4高變區(qū)擴(kuò)增和RLB分析。61份（59.80%）樣品用巴貝斯蟲／泰勒蟲屬特異性探針進(jìn)行雜交試驗(yàn),61份樣品中僅檢測到環(huán)形泰勒蟲。六份樣品只與泰勒蟲／巴貝斯蟲特異性探針雜交,而不與任何其他種屬特異性探針雜交,暗示有新的物種或基因型存在。對這6份樣品進(jìn)行18srRNA和ITS序列的擴(kuò)增,6份樣品的18S rRNA序列和ITS序列與環(huán)形泰勒蟲最接近... 

【文章來源】：華中農(nóng)業(yè)大學(xué)湖北省 211工程院校 教育部直屬院校

【文章頁數(shù)】：130 頁

【學(xué)位級別】：博士

【文章目錄】：
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


【參考文獻(xiàn)】：
期刊論文
[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]寄生于中國水牛Bubalus　Vbubalis的巴貝斯蟲一新種（梨形蟲目:巴貝斯科）[J]. 劉鐘靈,趙俊龍,馬麗華,姚寶安.  畜牧獸醫(yī)學(xué)報. 1997(01)



本文編號：3100974