我国犬埃立克体病研究现状

犬埃立克体病是由严格细胞内寄生的埃立克体所致的疾病。目前,我国已经发现的犬埃立克体病原有犬埃立克体(Ehrlichia)和扁平埃立克体(E.platy)两种。我国已发现的犬埃立克体病均为E.canis和E.platy的混合感染。临床症状包括发热、体重严重减轻、流鼻血和腹泻带血等;白细胞和血小板计数同时下降是我国犬埃立克体病的特点;严重的病犬常因广泛性出血或继发感染而死亡。犬埃立克体病的病原学诊断主要依靠镜检、血清学试验和PCR等。犬血清埃立克体抗体的调查结果表明,我国犬埃立克体病的流行与蜱活动密切相关。四环素类抗生素是治疗犬埃立克体病的有效药物。     

应用PCR检测犬埃立克体病的研究

用已确证为扁平埃立克体（Ehrlichia platys)和犬埃立克体（E.canis）混合感染的犬全血对6只格犬进行人工感染，通过对其临床症状，血流学变化，血涂片包涵体观察，并将其PCR结果与后来确诊为犬埃立克体病的18只病犬血的PCR结果比较，表明PCR方法能够对早期犬埃立克体病做出准确诊断。     

犬埃立克体实验模型初步研究

将犬埃立克体标准毒株（Florida株）体外细胞纯培养物和我国犬埃立克体病病犬血白细胞分别接种于C3H小鼠，接毒后d，经PCR检测证实均感染成功。本试验为犬埃立克体病小动物模型的建立打下了基础。     

犬在埃立克体病研究中的作用

随着新病原及病原新的致病性不断被发现 ,埃立克体及埃立克体病已经成为预防医学和兽医学研究热点。在已经发现的 1 1种埃立克体病原中 ,有 5种可以直接感染犬 ,引起不同的犬埃立克体病 ;另外 2种为早先分别从人单核细胞埃立克体病和粒细胞埃立克体病患者中分离的病原 ,在流行区的犬体内也可以分离到。资料表明犬对许多埃立克体都极为敏感 ,犬在埃立克体及埃立克体病研究上占有重要地位。鉴于这是我国近年才发现的新病原 ,许多兽医及研究工作者不认识它 ,文章就相关埃立克体对犬的感染和所致疾病以及在流行病学上的意义进行了综述。     

比格犬埃立克体病病理学研究

为了观察比格犬埃立克体病的病理学变化,用常规病理学方法,对6只人工感染埃立克体(Ehrlichia)的比格犬进行病理解剖学和病理组织学观察。结果:眼观所见最显著的病变是主要器官和皮下出血、淤血,脾、肝肿胀;病理组织学变化的主要特征是淋巴结的严重水肿,脾、肝、肺和肾的血管内及肝血管周围存在浆细胞和淋巴细胞灶性增生,皮肤表皮层变薄、皮肤皮脂腺腺泡上皮和毛囊变性坏死。比格犬埃立克体病的眼观和病理组织学变化与国外文献报道的基本吻合,不同之处是眼观病变更为明显,这可能与分离株的致病性不同有关。     

1例犬埃立克体病的诊治

犬埃立克体是一种重要的人兽共患传染性病原体,可引起犬科动物的犬单核细胞埃立克体病。为确诊1例临床上以发热、精神沉郁、黏膜苍白为主要特征的宠物犬病例,进行了血常规、血液生化检查、实时荧光定量PCR检测、犬埃立克体抗体检查,最终确诊该病例为犬埃立克体感染,使用盐酸多西环素治疗后患犬恢复良好。     

比格犬埃立克体病实验模型的建立

为犬埃立克体病的病理学、临床学、病原学的深入研究,我们用Beagle(比格犬）复制犬埃立克体病以建立犬埃立克体病实验模型。比格犬静脉接种E.platy和E.canis后,利用PCR和电镜技术确证感染成功。进一步的观察发现,接种后比格犬与最最感的德国牧羊犬自然感染埃立克体病的临床症关放病理学变化基本吻合。结果说明通过静脉接种病原成功建立了比格犬埃立克体病实验模型。     

比格犬人工感染埃立克体病的临床研究

为了表述比格犬感染埃立克体（Ehrlichia)后的临床表现，用已确证为Ｅ．platys和Ｅ．canis混合感染的犬血对５条比格犬进行了人工感染，并对临床和血液学变化进行了观察。埃立克体病的潜伏期为６－２９天，其后的临床表现为急性发热、眼结膜苍白、厌食、精神沉郁、体重严重减轻、腹泻带血、多处有淤血斑点出现、６－４５天内全部死亡。血液学变化显示血红蛋白和白细胞明显减少（Ｐ＜０．０１，Ｐ＜０．０５）。由此可见，比格犬对２种埃立克体（Ｅ.canis和Ｅ．platys）混合感染的性很高，严重的临床表现与美国学者普遍认为的亚临床状态相左，这可能是中国Ｇzh981株（Ｅ．platys)和Ｇzh982株（Ｅ．casnis)与国外株在毒力上的差异所致。     

Experimental transmission of granulocytic ehrlichial organisms in dogs

Canine granulocytic ehrlichial organisms were transmitted from an infected dog from Missouri to two male, 10-month-old dogs by an intravenous injection of whole blood. Physical or behavioral abnormalities were not detected during the 98 days of evaluation other than a mild pyrexia from Day 18 to 20. Ehrlichial morulae were found in blood granulocytes of Dog 1 from Day 13 to 44 and of Dog 2 from Day 14 to 34 with the peak rickettsemia occurring on Day 16 for both dogs. By Day 21 after inoculaiuon, both dogs had positive titers to Ehrlichia canis. The highest titers for both dogs were found 63 days after inoculation, after which the titers decreased. Most of the hematologic abnormalities (i.e., neutropenia, lymphocytosis, thrombocytopenia) and fever occurred between 18 and 24 days after inoculation. The pathologic bases of these abnormalities were not investigated but their concurrent presence suggested an association with the dogs' immunologic responses to the granulocytic ehrlichial agent. Results from the study indicated that the canine granulocytic ehrlichial agent of Missouri may produce subclinical infections and suggested that dogs may be able to clear the organism without antimicrobial therapy.     

Cultivation and molecular identification of Ehrlichia canis and Ehrlichia chaffeensis from a naturally co-infected dog in Venezuela

Background: Diagnosis of canine ehrlichiosis in Venezuela is normally performed by examination of buffy coat smears (BCS). Characteristic inclusion bodies are frequently observed in leukocytes and platelets from dogs with clinical signs of the disease. Objective: The purpose of this study was to investigate the co-infection of a dog with Ehrlichia canis and E hrlichia chaffeensis using microbiological and molecular techniques. Methods: Primary cultures of monocytes from a dog showing signs of ehrlichiosis were performed. Ehrlichial inclusions in blood cells were demonstrated by BCS and in cultured cell smears with direct immunofluorescence and Dip Quick staining. Nested PCR analysis was performed with DNA from blood samples and cultures, using primers specific for E. canis and E. chaffeensis. The amplified DNA fragments were sequenced to confirm the specificity of the amplifications. Results: The BCS of the naturally infected dog contained intracellular morulae. Ehrlichial inclusions were observed 9 days after inoculation of the primary cultures. After 3 passages with monocytes from a healthy dog, 65% of infected cells, and cells with >60 morulae were observed. A healthy female German Shepherd dog, seronegative for E. canis and E. chaffeensis antigens and without contact to ticks, was inoculated with an infected culture. The animal developed signs of canine monocytic ehrlichiosis and became seropositive. Nested PCR results and sequencing of amplified DNA fragments demonstrated the simultaneous presence of E. canis and E. chaffeensis in both dogs. Conclusions: This is the first report of E. chaffeensis in dogs in South America. This organism was previously identified in dogs by PCR only in the United States.     

同时检测犬瘟热病毒和犬细小病毒双重PCR方法的建立

为建立可以同时检测犬瘟热病毒(CDV)和犬细小病毒(CPV)的双重PCR方法,本研究根据GenBank登录的CDV N蛋白序列和CPV NS基因保守序列,设计合成2对特异性引物。通过优化反应条件,对CDV阳性病毒株反转录后的cDNA模板和CPV的DNA模板进行双重PCR扩增,同时得到2条与试验设计相符的669 bp(CDV)和392 bp(CPV)特异性条带,建立了同时检测CDV和CPV的双重PCR方法。实验结果表明:在同一PCR反应体系中可以同时检测这2种病毒,而对犬腺病毒Ⅰ型、犬腺病毒Ⅱ型、狂犬病毒检测均为阴性;CDV和CPV的最低检出限分别为101.8TCID50和101.4TCID50。采用该方法对在黑龙江省不同地区所采集的30份犬病料样品进行检测,CDV阳性率为30%;CPV阳性率为23.33%,表明建立的PCR方法可以用于临床诊断。     

犬源牛犬细小病毒和犬圆环病毒二联PCR检测方法的建立及应用

本研究旨在建立可同时检测犬源牛犬细小病毒(CBoV)和犬圆环病毒(CCV)的二联PCR检测方法,并对两种病毒病当前的流行情况进行监测和调查.分别将已发表的CBoV和CCV基因组序列进行同源性比对,选择高同源区段,应用Primer Primier 5计算机软件设计并合成了2对特异性扩增引物,目的片段大小分别为170 bp...