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1.
以广州某宠物医院临床犬源贾第虫为研究对象,根据GenBank TM中登录的贾第虫EF1α基因序列(AF069575),设计1对引物EF/ER,采用PCR方法从贾第虫基因组DNA中扩增出目的片段,纯化后克隆至pGEM-T Easy载体,重组质粒通过菌液PCR鉴定后,对其进行序列测定及分析,旨在对广州犬源贾第虫进行分子鉴定。结果显示,该犬源贾第虫EF1α序列长为288bp,与预期目的片段一致,该序列构建的分子进化树表明该犬源贾第虫分离株为蓝氏贾第虫D型。说明EF1α适合做分子标记,可准确地对贾第虫进行基因分型,为贾第虫分子分类学及分子遗传学研究奠定了基础。 相似文献
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犬贾第虫dsRNA病毒的鉴定及特性 总被引：12，自引：0，他引：12
对分离到的6株犬贾第虫（Giardia cnais)包囊进行核酸分析，在其中1株犬贾第虫核酸的琼脂糖凝胶电泳图谱上观察到1条长度约7.0kb的片段。经鉴定，该核酸不能被DNA酶（100mg/L）降解，对质量浓度低的RNA酶（0.1mg/L)不敏感，但可被质量浓度高的RNA酶（10mg/L）降解。纯化包囊经流氮冻融后，磷钨酸负染，电镜观察发现有球形、直径约为33nm的病毒样粒子存在，包囊超薄切片在胞质中也观察到该病毒样粒子存在。RNA依赖RNA聚合酶活性测定结果表明，该病毒具有RNA依赖RNA聚合酶的活性。犬贾第虫dsRNA病毒核经RT-PCR扩增后得到1条预计扩增大小的片段，将其回收后连接到pMD18-T载体上进行克隆并测序，所得序列仅与蓝氏贾第虫病毒具有较高同源性。 相似文献
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从吉林省某犬繁育中心自然感染的比格犬中分离到一株贾第虫,经鉴定其为贾第虫属蓝氏贾第虫种犬贾第虫亚种,本研究提供了一套简便可行的犬贾第虫包囊的分离的分离纯化方法,将有利于对该病的进一步研究. 相似文献
4.
采用巢式PCR鉴定广东两株犬源贾第虫(GD1和GD2)的基因型。运用碘液染色对犬粪便中贾第虫进行鉴定,提取DNA后经巢式PCR扩增tpi基因,扩增产物测序后用BLAST和DNAStar软件进行同源性和系统发育分析。结果表明,通过tpi基因分析,GD1与L02120的同源性为100%,种系发育树上位于同一分支;GD2与DQ220289的同源性为99.1％,种系发育树上两者在同一分支。广东犬源贾第虫GD1为人畜共患的A1型,GD2为犬的D型。 相似文献
5.
为研究青海省海北地区牦牛贾第虫的感染情况及虫种基因型,对青海省祁连县、海晏县和刚察县的297份牦牛粪样采用蔗糖密度梯度离心法纯化,之后用免疫荧光方法对贾第虫进行鉴定,对阳性及疑似阳性样品采用基于18SrRNA和谷氨酸脱氢酶(gdh)基因的套式PCR扩增,并对扩增产物进行测序。将测序结果与GenBank中的贾第虫序列进行比对分析。免疫荧光抗体试验结果显示,共检出24份贾第虫阳性粪样,总阴性率为8.1%。套式PCR扩增结果显示,24份阳性样品中18SrRNA基因扩增阳性22份,gdh基因扩增阳性18份,产物大小分别为292bp和432bp。序列分析表明,分离的虫种均为牦牛源肠贾第虫,基因型为集聚体E,未发现人畜共患基因型。 相似文献
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中国人源蓝氏贾第虫病毒基因组全长cDNA克隆及序列测定 总被引：4，自引：0，他引：4
从中国人源蓝氏贾第虫北京株(BEIJ88／BTMR1／1)体外纯培养的电镜负染和超薄切片观察到蓝氏贾第虫病毒粒子。该病毒位于感染早期贾第虫的细胞质中，感染晚期的细胞核中。根据国外发表的蓝氏贾第虫病毒序列设计了6对互相重叠的引物。用这6对引物对从中国人源蓝氏贾第虫北京株发现的蓝氏贾第虫病毒基因组进行RT—PcR，将产物连接到pMD18-T载体中并转化入DH5a感受态菌，送上海生工测序，通过BLAST对Gen-Bank进行同源性搜索。结果 测得我国人源蓝氏贾第虫病毒基目组全长为6273bp，与国外报道的蓝氏贾第虫病毒序列同源性为95％，编码的氨基酸同源性为90％。 相似文献
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犬贾第虫病毒(长春株)全基因组序列分析 总被引：2，自引：0，他引：2
根据已报道的人源蓝氏贾第虫病毒序列设计了互相重叠的6对特异性引物，以犬贾第虫（长春株）纯培养滋养体总核酸为模板进行RT—PCR。PCR产物连接到pMD18-T载体进行测序，通过BLAST在GenBank进行同源性搜索，并用DNAMAN分子生物学软件进行分析。结果测得我国犬贾第虫病毒（长春株）基因组全长为6276bp（DQ238861），编码1个有887个氨基酸残基的衣壳蛋白和1056个氨基酸残基的融合蛋白，这2个阅读框被-1核糖体移码框分开，重叠处有220nt。基因组中G＋C占49．62％。其序列与国外报道的人源蓝氏贾第虫病毒（L13218）序列同源性为94．62％，编码的氨基酸同源性为93．50％；与国内人源蓝氏贾第虫病毒（AF525216）序列同源性为98．88％，编码的氨基酸同源性为98．30％。 相似文献
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解放军军需大学军事兽医系军事预防兽医学教研室张西臣教图 1 中国人源蓝氏贾第虫病毒负染电镜照片 ( 132 0 0 0× ) 球形病毒粒子中发白的为完整病毒 ,发黑的为病毒衣壳授领导的课题组最近又有重大发现 ,首次在我国蓝氏贾第虫(Giardia lambia)体内发现病毒粒子。试验用 TYI- S- 33培养基对中国人源蓝氏贾第虫北京株 (BEIJ88/BTMR1/1)进行体外纯培养 ,将培养液上清 45 0 0 0 r/min 4℃离心 2 h,后取沉淀用磷钨酸负染 ,并作超薄切片 ,进行电镜观察 ,同时对蓝氏贾第虫病毒核酸进行了分析 ,测定了病毒全长基因组序列。结果发现中国人源… 相似文献
10.
从青海省祁连县采集93份牦牛粪样用于贾第虫检测。所有样品经蔗糖密度梯度离心法纯化处理后,采用卢戈氏碘液染色镜检和免疫荧光试验进行检测,将镜检阳性样品采用套式PCR方法扩增18S rRNA,并对扩增产物进行测序分析。结果镜检和免疫荧光试验中有9份样品为贾第虫阳性,阳性率为9.7%。镜检阳性样品采用套式PCR方法扩增后有8份样品为18SrRNA基因阳性,产物长度292bp,测序后的序列分别命名为QHQL201501~QHQL201508。系统发育分析显示,分离的虫株属于牛源十二指肠贾第虫,基因型均为反刍动物特有的聚集体E,未发现具有人兽共患潜力的A型和B型。 相似文献
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Scorza AV Ballweber LR Tangtrongsup S Panuska C Lappin MR 《Veterinary parasitology》2012,189(2-4):182-188
The objective of this study was to determine and compare the assemblages of Giardia duodenalis isolated from mammalian fecal samples using the β-giardin (bg), glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi) genes. A total of 202 samples, either submitted to the Veterinary Diagnostic Laboratory (Parasitology) at Colorado State University or part of ongoing research studies, were typed. A subset of 50 dog samples were also assessed by the tpi-D-specific primers. Of these, 183 were from dogs, 13 were from cats, two were from llamas, and one each was from a calf, an alpaca, a sheep, and a horse. The majority of the dogs (171 of 183 isolates) in this study were infected with only dog-adapted Assemblage C or D. The tpi-D-specific primers confirmed that 28 of the samples that typed as Assemblage D by the bg and gdh genes were also Assemblage D by the tpi-D-specific primers. Only 12 isolates were Assemblage A alone or Assemblage A and Assemblage C or D. Of the 13 cat isolates, seven were Assemblage F, two were Assemblage D, three were Assemblage A and 1 contained both Assemblages C and D. The calf isolate was Assemblage E (gdh, tpi) and the alpaca (bg, gdh), llamas (gdh), sheep (bg, gdh, tpi) and horse (tpi) isolates were all Assemblage A. When the assemblage could be determined for more than one gene, 91 of 117 dog isolates gave consistent results and 8 of 9 cat isolates gave consistent results. 相似文献
12.
IntroductionCanine dilated cardiomyopathy (DCM) can result from numerous etiologies including genetic mutations, infections, toxins, and nutritional imbalances. This study sought to characterize differences in echocardiographic findings between dogs with DCM fed grain-free (GF) diets and grain-based (GB) diets.AnimalsForty-eight dogs with DCM and known diet history.MethodsThis was a retrospective analysis of dogs with DCM from January 1, 2015 to May 1, 2018 with a known diet history. Dogs were grouped by diet (GF and GB), and the GF group was further divided into dogs eating the most common grain-free diet (GF-1) and other grain-free diets (GF-o). Demographics, diet history, echocardiographic parameters, taurine concentrations, and vertebral heart scale were compared between GB, all GF, GF-1, and GF-o groups at diagnosis and recheck.ResultsDogs eating GF-1 weighed less than GB and GF-o dogs, but age and sex were not different between groups. Left ventricular size in diastole and systole was greater, and sphericity index was less for GF-1 compared with GB dogs. Diastolic left ventricular size was greater for all GF compared with that of GB dogs. Fractional shortening, left atrial size, and vertebral heart scale were not different between groups. Taurine deficiency was not identified in GF dogs, and presence of congestive heart failure was not different between groups. Seven dogs that were reevaluated after diet change (6 received taurine supplementation) had clinical and echocardiographic improvement.ConclusionsDietary-associated DCM occurs with some GF diets and can improve with nutritional management, including diet change. The role of taurine supplementation, even without deficiency, is uncertain. 相似文献
13.
Babesia canis has generally been considered the only large Babesia to infect dogs. In this study, we used PCR to detect and characterize B. canis canis isolated from naturally infected dogs in Poland by amplifying and sequencing a portion of the 18S ribosomal RNA (rRNA) gene. Venous blood samples were collected from 76 Babesia-symptomatic dogs. A 559-bp fragment of the B. canis canis 18S rRNA gene was amplified by PCR. The PCR products were then digested with HincII restriction enzyme, and isolates were classified according to whether they were cut (group A) or not (group B) by this endonuclease. Sequencing of the PCR products from the isolates led to the identification of seven sequence variants (four in group A, and three in group B). Sequences were compared with GenBank sequences, and alignments showed that all B. canis canis isolates from Europe may be classified into groups A or B as defined in our study. 相似文献
14.
Quantification of mucin gene expression in tracheobronchial epithelium of healthy dogs and dogs with chronic bronchitis 总被引：1，自引：0，他引：1
Hawkins EC Birkenheuer AJ Marr HS Rogala AR Large EE Adler KB 《American journal of veterinary research》2007,68(4):435-440
OBJECTIVE: To develop a real-time PCR assay for the quantification of mucin gene expression in tracheobronchial brushing specimens from dogs and compare mucin gene expression in specimens from dogs with naturally occurring chronic bronchitis with that in specimens from healthy dogs. ANIMALS: 7 healthy dogs and 5 dogs with chronic bronchitis. PROCEDURES: Primers that were designed to span the predicted intron-exon boundaries of a canine MUC5AC-like gene were used to develop a real-time PCR assay for quantification of expression of that gene. Total mRNA was isolated from tracheobronchial brushing specimens obtained from dogs with and without bronchitis during anesthesia; MUC5AC-like gene expression in those samples was quantified by use of the real-time PCR assay. RESULTS: The PCR assay was sensitive and specific for the target sequence, the predicted amino acid sequence of which had greatest homology with human, porcine, and rat MUC5AC. The assay was able to quantify the target over a wide dynamic range. Dogs with chronic bronchitis had a 3.0-fold increase in the quantity of MUC5AC-like mRNA, compared with healthy dogs. CONCLUSIONS AND CLINICAL RELEVANCE: The ability to measure mucin gene expression from tracheobronchial brushing specimens collected from client-owned dogs during routine bronchoscopy should prove to be a useful tool for the study of bronchitis in dogs and expand the usefulness of airway inflammation in dogs as a model for bronchitis in humans. 相似文献
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Presence of human Giardia in domestic,farm and wild animals,and environmental samples suggests a zoonotic potential for giardiasis 总被引：2，自引：0，他引：2
van Keulen H Macechko PT Wade S Schaaf S Wallis PM Erlandsen SL 《Veterinary parasitology》2002,108(2):97-107
Giardia lamblia which parasitize humans belong to either of two genotypes, A or B, based on specific signature sequences in the 5' end of the small subunit (16S) ribosomal RNA (rRNA) gene. These two genotypes also were found in cysts from fecal samples of animal origin such as dogs, cats, some farm animals and wild animals. In addition, trophozoites recovered from cysts obtained from environmental samples belonged to these two genotypes as well, suggesting that the G. lamblia genotypes A and B are widespread and possibly zoonotic. Trophozoites were recovered from rats and these isolates might belong to another genotype of G. lamblia. Deer mice and one dog appeared to be parasitized by genotypes of Giardia with close affinity to G. microti. This species, therefore, also consists of a genotype complex. 相似文献
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Solano-Gallego L Rodriguez-Cortes A Trotta M Zampieron C Razia L Furlanello T Caldin M Roura X Alberola J 《Veterinary parasitology》2007,147(3-4):315-319
The aim of this study was to detect Leishmania infantum DNA by real-time PCR in urine from different groups of dogs with clinical leishmaniosis. Urine from 10 clinically healthy dogs and 43 dogs with clinical leishmaniosis diagnosed by positive serology and/or bone marrow PCR were studied. The group of 43 dogs with clinical leishmaniosis was divided into three subgroups: 13 dogs with renal insufficiency and proteinuria (urine protein-creatinine ratio greater than one), 13 dogs with only proteinuria, and 17 dogs with neither renal insufficiency nor proteinuria. The detection of Leishmania DNA was performed by light cycler real-time PCR using hybridization probes in each urine sample. Leishmania positive PCR was found in 47% (20/43) of the urine from leishmaniotic dogs, while all urine from clinically healthy dogs were negative. The percentages of positive Leishmania PCR were 85% (11/13) in dogs with renal insufficiency and proteinuria, 23% (3/13) in dogs with proteinuria and 35% (6/17) in dogs with neither renal insufficiency nor proteinuria. Dogs with renal insufficiency and proteinuria presented a statistical significant greater percentage of positive Leishmania PCR in urine when compared with the other subgroups (P<0.02). This study demonstrates the presence of Leishmania DNA in urine of dogs with leishmaniosis. Those dogs with severe renal damage present a greater number of Leishmania parasites in urine. 相似文献
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石河子地区宠物犬蜱的种类鉴定及其携带布鲁氏菌的检测 总被引：1，自引：0，他引：1
为了解新疆石河子地区宠物犬体表寄生蜱的种类及蜱携带布鲁氏菌情况,本试验在形态学分类的基础上,基于线粒体基因12S rRNA和细胞色素氧化酶Ⅰ（COⅠ）对宠物犬体表采集寄生蜱进行分子生物学检测,使用DNAMAN软件比对分析序列同源性,并应用序列分析软件Mega 7.0邻接法构建蜱种系统发育进化树,分析蜱种的遗传进化关系;基于布鲁氏菌外膜蛋白Omp22基因对宠物犬体表寄生蜱进行布鲁氏菌PCR检测,确定宠物犬蜱布鲁氏菌携带情况。结果显示,宠物犬体表寄生的436只蜱的形态学与线粒体基因（12S rRNA和COⅠ）分子生物学鉴定结果一致,均为新疆优势蜱种之一——图兰扇头蜱（Rhipicephalus turanicus）。基于12S rRNA基因构建的蜱种系统发育树显示,本试验所得宠物犬体表寄生图兰扇头蜱序列与GenBank中已知图兰扇头蜱的序列聚为一大支。基于布鲁氏菌Omp22基因的巢式PCR扩增结果显示,宠物犬体表寄生图兰扇头蜱携带布鲁氏菌的阳性率为4.82%（21/436）,且同源性为100%。BLAST比对显示,本试验所得宠物犬图兰扇头蜱携带的布鲁氏菌与新疆流行株YC31（GenBank登录号：MK201679.1）、QH5（GenBank登录号：MK201678.1）、EM3（GenBank登录号：MK201677.1）和ML9（GenBank登录号：MK201676.1）的同源性均为100%。本研究通过形态学及分子生物学探讨了新疆石河子地区宠物犬体表寄生蜱的种类及携带布鲁氏菌基本情况,为宠物犬体表寄生蜱的种类及蜱传疾病的监测和控制等研究工作提供了基础资料。 相似文献
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IntroductionDilated cardiomyopathy (DCM) in dogs has been associated with feeding of grain-free (GF), legume-rich diets. Some dogs with presumed diet-associated DCM have shown improved myocardial function and clinical outcomes following a change in diet and standard medical therapy.HypothesisPrior GF (pGF) diet influences reverse cardiac remodeling and clinical outcomes in dogs with DCM and congestive heart failure (CHF).Animals and methodsA retrospective study was performed with 67 dogs with DCM and CHF for which diet history was known. Dogs were grouped by diet into pGF and grain-inclusive (GI) groups. Dogs in the pGF group were included if diet change was a component of therapy. Survival was analyzed using Kaplan–Meier curves and the Cox proportional-hazards model.ResultsThe median survival time was 344 days for pGF dogs vs. 253 days for GI dogs (P = 0.074). Statistically significant differences in median survival were identified when the analysis was limited to dogs surviving longer than one week (P = 0.033). Prior GF dogs had a significantly worse outcome the longer a GF diet was fed prior to diagnosis (P = 0.004) or if they were diagnosed at a younger age (P = 0.017). Prior GF dogs showed significantly greater improvement in normalized left ventricular internal diastolic diameter (P = 0.038) and E-point septal separation (P = 0.031) measurements and significant decreases in their furosemide (P = 0.009) and pimobendan (P < 0.005) dosages over time compared to GI dogs.ConclusionsPrior GF dogs that survived at least one week after diagnosis of DCM, treatment of CHF, and diet change had better clinical outcomes and showed reverse ventricular remodeling compared to GI dogs. 相似文献