Molecular Seroepidemiologic Studies on Piroplasmosis of Domestic Animals in Brazil

巴西家畜焦虫病分子血清流行病学研究

• 批准号: 12575028
• 负责人: FUJISAKI Kozo
• 金额: $ 9.22万
• 依托单位: Obihiro University of Agriculture and Veterinary Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12575028/
• 关键词: piroplasmosis; Theileria orientalis sergenti; herpes virus; recombinant vaccine; Babesia parasites; gene cloning; vector tick; Haemaphysalis longicornis; ピロプラズマ病; タイレリア原虫; バベシア原虫; 血清抗体; 媒介ダニ; DNA診断; 住血原虫病; ELISA; カピパラ; トリパノソーマ

(1) Serum samples from horses in the States of Sao Paulo and Mato Grosso do Sul, Brazil were examined for diagnosis of equine piroplasmosis by both the latex agglutination test (LAT) and the enzyme-linked immunosorbent assay (ELIZA) with recombinant antigens. Of the examined samples, 81% and 90% samples were positive for Babesia equi and Babesia caballi infection, respectively. Also, 75%75% samples were positive for both B. equi and B. caballi mixed infections. These results indicate that equine piroplasmosis is widespread and therefore a cause for serious concern in the States of Sao Paulo and Mato Grosso do Sul, Brazil.(2) The potential role of Boophilus microplus as natural tick vector of B. equi and B. caballi in Brazilian horses was assessed using nested polymerase chain reaction (PCR)-based marker assay. B. equi merozoite-specific 218bp gene fragment (EMA-1) was detected in 45.3-62.5% of females, eggs, larvae, and nymphs of B. microplus collected from 47 horses at the State of Ma … More to Grosso do Sul, Brazil.Except for the partially-fed female ticks, the B. caballi-specific 430 bp gene fragment was amplified from developmental stages. Therefore, we have obtained molecular evidence that strengthens earlier experimental and ultrastructural studies on Brazil incriminating B. microplus as a natural vector of B. equi, and possibly of B. caballi.(3) Primers specific for EMA-1 gene of B. equi successfully amplified products from all samples of Dermacentor nuttalli adult ticks from Mongolia while primers for the 430 bp gene fragment product from BC48 gene of B. caballi amplified products from seven of the 54 samples. We have found mixed infections with B. equi and B. caballi in the tick vector. These results suggest that D. nuttalli may play an important role as a vector of both B. caballi and B. equi and also may be important in maintenance of endemicity of equine piroplasmosis in Mongolia. The detection of parasite DNA in eggs and larval progenies is likewise suggestive of transovarial parasite transmission in D. nuttalli ticks.(4) At least three different variations in EMA-1 gene of B. equi strains isolated from different regions. The phylogenetic tree deduced from the sequences suggests a closer relationship between the Brazilian and Florida strains of B. equi DNA, and a distant relationship between B. equi DNA isolated from the Brazilian samples and USDA strain. B. equi gene obtained from Mongolian samples revealed comparatively distant relationship to other examined strains. Less

(1)用重组抗原乳胶凝集试验(LAT)和酶联免疫吸附试验(ELIZA)检测了巴西圣保罗州和南马托格罗索州马匹的血清样本。检测样本中，马巴贝斯虫感染阳性率为81%，卡巴贝斯虫感染阳性率为90%。同时，75%-75%的样本同时检测到马巴氏杆菌和卡巴利巴氏杆菌混合感染。这些结果表明，马的梨形体病在巴西的圣保罗州和南马托格罗索州非常普遍，因此引起了人们的严重关注。(2)采用基于巢式聚合酶链式反应(PCR)的标记分析方法，评估了微小牛瘟杆菌作为马巴斯虫和卡巴利虫的天然媒介在巴西马中的潜在作用。在马…州采集的47匹马的雌虫、卵、幼虫和若虫中，检测到45.3-62.5%的马裂殖子特异的218bp基因片段(EMA-1)巴西南格罗索州。除了部分喂食的雌蚤外，从发育阶段扩增出了卡巴利绦虫特异的430个碱基基因片段。因此，我们获得了分子证据，加强了早期关于巴西微小乳杆菌可能是马血吸虫和卡巴利血吸虫的天然载体的实验和超微结构研究。(3)针对马巴氏血吸虫EMA-1基因的特异引物成功地扩增了蒙古国所有样本的纳氏革螨成虫的扩增产物，而从54个样本中有7个样本扩增了巴氏血吸虫BC48基因的430bp的基因片段产物。我们在硬蜱病媒中发现了马巴氏杆菌和卡巴利杆菌的混合感染。这些结果表明，纳氏丝虫可能作为卡巴利支原体和马支原体的媒介发挥重要作用，也可能对维持蒙古国马匹罗支原体病的流行起重要作用。在虫卵和幼虫后代中检测到寄生虫DNA也同样提示了经卵传播的寄生虫。(4)不同地区分离到的马分支杆菌EMA-1基因至少有三个不同的变异。根据这些序列构建的系统发育树表明，巴西株和佛罗里达株之间有较近的亲缘关系，而从巴西样本中分离到的马分支杆菌DNA与美国农业部的株之间有较远的亲缘关系。从蒙古族样品中获得的马不动杆菌基因与其他受检菌株具有较远的亲缘关系。较少

项目成果

Xuan, X.: "Diagnosis of equine piroplasmosis in Xinjiang province of China by the enzyme-linked imunosorbent assays using recombinant antigens"Vet.Parasitol.. 108・2. 179-182 (2002)

宣X.：“通过使用重组抗原的酶联免疫吸附测定诊断中国新疆省的马焦虫病”Vet.Parasitol.. 108・2（2002）

Battsetseg, B., et al.: "Detection of Babesia caballi and Babesia equi in Dermacentor nuttalli adult ticks"Int. J. Parasitol.. 31・4. 383-386 (2001)

Battsetseg, B., et al.：“Dermacentor nuttalli 成蜱中的巴贝斯虫和马巴贝斯虫的检测”Int. J. Parasitol.. 31・4 (2001)。

Xuan, X.: "Detection of antibodies to Babesia equi in horses by a latex agglutination test using recombinant EMA-1"Clin. Diagn. Lab. Immun. 8・3. 645-646 (2001)

Xu，X.：“使用重组 EMA-1 进行乳胶凝集试验检测马巴贝斯虫抗体”Clin. Immun. 8·3 (2001)。

Battsetseg, B.: "Diagnosis of Babesia caballi and Babesia equi infections in Dermacentor nuttalli adult ticks"Int. J. Parasitol.. 31・4. 384-386 (2001)

Battsetseg，B.：“Dermacentor nuttalli 成年蜱中的巴贝斯虫和马巴贝斯虫感染的诊断” Int. J. Parasitol.. 31・4 (2001)。

Inoue,N.: "Novel species specific antigens of Trypanosoma congolense and their different localization among life-cycle stages. "Journal of Veterinary Medical Science. 62・10. 1041-1045 (2000)

Inoue, N.：“刚果锥虫的新物种特异性抗原及其在生命周期阶段的不同定位。”兽医医学杂志 62・10（2000）。