首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 46 毫秒
1.
本试验采用Vero细胞从临床疑似患犬瘟热的犬组织病料中分离出一株病毒,通过观察细胞病变,提取细胞培养物RNA并扩增克隆分析N蛋白基因部分序列进行了鉴定。结果表明,该毒株接种于Vero细胞后,出现了典型的细胞病变;对接种病料后的Vero细胞培养物提取总RNA进行RT-PCR扩增,得到了287 bp的目的片段;序列分析表明该分离株与犬瘟热病毒弱毒(Onderstepoort和Snyder Hill)的同源性为95.5%~96.9%,与标准强毒株(A75/17和2544/Han95)及野毒株(XJ-4、ZJ7、98-2646等)的同源性较高,为97.2%~99.0%,证明该毒株为犬瘟热病毒野毒株,命名为QD10。  相似文献   

2.
为构建一株表达狂犬病病毒(RV) G蛋白的重组犬瘟热病毒(CDV),本研究以本实验室分离的CDVSnyder Hill疫苗株为基础,在CDV P和M基因之间插入RV弱毒疫苗株ERA G蛋白基因,构建重组质粒p CI-CDV-RVG及表达CDV N、P和L蛋白的辅助质粒p CI-CDVN、p CI-CDVP和p CI-CDVL,将重组质粒和辅助质粒共转染BSR细胞,拯救获得了表达RV G蛋白的重组犬瘟热病毒(rCDV-RVG),将该重组病毒在Vero细胞中传代,通过RT-PCR、间接免疫荧光试验(IFA)和western blot鉴定。结果显示RV G基因能够在r CDV-RVG中稳定存在并正确表达。病毒生长动力学曲线显示,重组病毒在Vero细胞中的增殖效率与拯救的亲本病毒r CDV无显著差异(p0.05)。本研究表明CDV Snyder Hill株作为载体表达外源基因的潜力,为研制高效、安全、廉价的CDV-RV二联活载体疫苗奠定基础。  相似文献   

3.
根据已发表的犬瘟热病毒OP株核酸序列设计2对引物,以犬瘟热病毒OP株感染Vero细胞收获病毒提取的RNA为模板,应用RT—PCR扩增出F基因的769、832bp片段。将PCR产物按正确的阅读框架定向克隆进pGKX—6p—1载体中谷胱甘肽—S—转移酶(GST)基因的下游,再将重组质粒转化入宿主菌BL21中,在37℃ 1.0mmo1/L IPTG诱导下,F基因2个片段获得了良好的表达。表达产物经聚丙烯酰胺凝胶电泳鉴定,确定所表达的融合多肽大小分别为54000、56000。将表达产物回收后混合免疫小鼠,IFA显示,免疫小鼠血清能与病毒感染细胞呈现特异反应,并表现出1:16的中和抗体活性,表明体外表达的F多肽保留了天然蛋白的部分抗原性。  相似文献   

4.
貉源犬瘟热病毒的分离鉴定   总被引:1,自引:1,他引:0  
采集大庆某貉养殖场病貉的病料,处理后接种于非洲绿猴肾细胞(Vero)进行病毒分离。对分离毒株进行了中和试验、血凝试验、理化性质的鉴定,并用反转录-聚合酶链式反应(RT-PCR)检测感染细胞中的病毒核酸。病料接种 Vero 细胞 72 h 后产生明显的细胞病变(CPE);病毒分离株可以被犬瘟热病毒阳性血清中和,中和效价为 1∶25;分离株对氯仿、乙醚敏感,对酸和热抵抗力弱,可以凝集鸡红细胞,其凝集作用可被犬瘟热病毒阳性血清所抑制; RT-PCR检测病毒细胞培养液,扩增出的片段长287 bp,与预期设计的长度相同,经测序发现与MS01株的同源性为99%。结果表明,分离的病毒株为犬瘟热病毒,命名为CDV-DQ株。  相似文献   

5.
旨在构建稳定表达SLAM受体的SLAM-Vero细胞系和SLAM-BHK21细胞系,并比较其对犬瘟热病毒(CDV)野毒株的敏感性,为CDV野毒株的快速分离与研究提供一种有效的工具。将真核表达载体Pcag-SLAM分别转染Vero细胞和BHK21细胞。经G418压力筛选结合有限稀释法筛选阳性克隆株,并通过RT-PCR、间接免疫荧光鉴定(IFA)和Western blot等方法对稳定表达SLAM受体的SLAM-Vero细胞系和SLAM-BHK21细胞系加以验证。应用这两种细胞系对3例临床犬瘟热病例的5种不同组织进行病毒分离,对分离得到的CDV进行RT-PCR及IFA鉴定。结果显示,经RT-PCR和Western blot验证,本研究成功构建出SLAM-Vero细胞系和SLAM-BHK21细胞系,SLAM-Vero细胞接种病毒12~24h即可出现典型的CDV导致的细胞融合体CPE,利用SLAM-Vero细胞从3只CDV阳性犬的肺和脾中分离得到2株CDV,而SLAM-BHK21细胞、亲本Vero细胞及亲本BHK21细胞未能分离出病毒。研究表明,与SLAM-BHK21细胞相比,SLAM-Vero细胞对CDV的分离率较高,并能产生明显的CPE。在分离CDV野毒株时,选择肺和脾更容易在SLAM-Vero细胞系上分离出病毒。  相似文献   

6.
试验比较了MA-104、Marc-145、Vero 3种猴肾细胞增殖犬瘟热病毒的敏感性,按常规方法将犬瘟热病毒(CDV-XN112株)分别接种于MA-104、Marc-145、Vero 3种细胞,通过观察细胞病变,确定收毒时间,比较病毒滴度。结果表明,犬瘟热病毒可在MA-104、Marc-145、Vero 3种细胞内增殖,并出现典型细胞病变,效价均达到104.5TCID50/0.1mL以上,而Vero细胞增殖犬瘟热病毒更为经济高效,是增殖该病毒的理想细胞。  相似文献   

7.
根据已发表的犬瘟热病毒(CDV)Onderstepoort株的序列设计两对引物,以犬瘟热病毒云南株感染的Vero细胞收获病毒提取的总RNA为模板,RT-PCR扩增出H基因的939 bp(H基因前段,H1)和920 bp(H基因后段,H2)片段,分别将其定向克隆于PET-28a( )中,将重组质粒转化宿主菌BL21(DE3)plysS,在35℃1、.0 mmol/L IPTG诱导下获得良好表达,经SDS-PAGE鉴定,表达的融合蛋白为34 ku,与预期大小一致。免疫印迹试验显示,该重组蛋白可被CDV Onderstepoort株多克隆抗体识别,表明该重组蛋白具有抗原性。  相似文献   

8.
将犬瘟热病毒(Canine distemper virus,CDV)疫苗株接种Vero细胞,连续培养至TCID50为10-4.625/m L。采用肋间肌多点注射方法免疫马,4周后每2周采集血液,分离血清,病毒中和试验测定血清中中和抗体效价,最高值达到212.5。以硫酸铵沉淀法纯化Ig G,病毒中和试验测定其中和抗体效价可达214。通过肌注犬观察,无明显不良反应。研究结果证明,本试验制备的马源免疫球蛋白可用于犬的抗体治疗,为犬瘟热的治疗制剂研发开辟了新方向。  相似文献   

9.
用919株牛暂时热病毒制造细胞培养疫苗。Tzipori(1974)曾将其他牛暂时热BEF病毒株适应于Vero细胞系,并在啮齿动物进行传代,结果传代毒对牛的抗原性不如919株好。919株弱毒是将病牛血毒直接接种Veto细胞,因而避免了病毒抗原性发生改变。将919株弱毒经静脉接种犊牛所诱发的中和抗体应答和犊牛被接种417WBC株强毒后发  相似文献   

10.
为了研究当前犬瘟热的流行毒株,试验选取来自辽宁省大连市疑似犬瘟热发病死亡水貂的肺脏、脾脏、肾脏等组织,将研磨匀浆后的上清液接种于稳定表达犬瘟热SLAM受体的非洲猕猴肾细胞系-Vero/DogSLAM(VDS),通过病料的处理、VDS细胞的复苏与培养、病毒的分离与培养、病毒TCID_(50)的测定、反转录-聚合酶链式反应(RT-PCR)的方法检测感染细胞中的病毒核酸,用序列比对的方法与已发表的犬瘟热病毒H蛋白(CDV-H)和SLAM受体结合区(RBS)序列进行比对分析,用CDV-H蛋白间接免疫荧光(IFA)的方法检测荧光蛋白表达。结果表明:在Vero/DogSLAM(VDS)细胞上接种病毒培养24 h后,80%~90%的VDS细胞出现合胞体病变(CPE),分离到的病毒TCID_(50)达到1×10~(5. 23)/(100μL),同时检测的CDV-H和SLAM受体结合区(RBS)序列比对分析一致,同源性为98%以上,在细胞质中可观察到抗CDV-H蛋白的特异性绿色荧光,RT-PCR成功扩增出H基因部分片段。说明本试验成功分离并鉴定出1株犬瘟热毒株,将其命名为LNDL(17) M4株。  相似文献   

11.
1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0-9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks.  相似文献   

12.
试验选用96头平均体重14.82 kg左右的杜×长×大断奶仔猪,随机分成4组,每组3栏,每栏8头(公母各半)。对照组饲喂基础日粮,试验1、2、3组分别添加1%80目白术、0.2%白术多糖和1%微米白术。试验期30 d。结果表明:在生长性能方面,与对照组相比,1%微米白术添加组可显著提高日增重(P0.05)、降低饲料增重比和腹泻率,而且效果优于1%80目白术组和0.2%白术多糖组,在肠道形态和肠道微生态区系方面,与对照组相比,日粮添加1%80目白术、0.2%白术多糖、1%微米白术均可不同程度的提高十二指肠和空肠的绒毛高度,加深十二指肠和空肠的隐窝深度,并且增加肠道微生态区系的多样性,其中以1%微米白术添加组的效果最佳。  相似文献   

13.
Sissay, M.M., Uggla, A. and Waller, P.J., XXXX. Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia. Tropical Animal Health and Production, XXXX. A 2-year abattoir survey was carried out to determine the prevalence, abundance and seasonal incidence of gastro-intestinal (GI) nematodes and trematodes (flukes) of sheep and goats in the semi-arid zone of eastern Ethiopia. During May 2003 to April 2005, viscera including liver, lungs and GI tracts were collected from 655 sheep and 632 goats slaughtered at 4 abattoirs located in the towns of Haramaya, Harar, Dire Dawa and Jijiga in eastern Ethiopia. All animals were raised in the farming areas located within the community boundaries for each town. Collected materials were transported within 24 h to the parasitology laboratory of Haramaya University for immediate processing. Thirteen species belonging to 9 genera of GI nematodes (Haemonchus contortus, Trichostrongylus axei, T. colubriformis, T. vitrinus, Nematodirus filicollis, N. spathiger, Oesophagostomum columbianum, O. venulosum, Strongyloides papillosus, Bunostomum trigonocephalum, Trichuris ovis, Cooperia curticei and Chabertia ovina), and 4 species belonging to 3 genera of trematodes (Fasciola hepatica, F. gigantica, Paramphistomum {Calicohoron} microbothrium and Dicrocoelium dendriticum) were recorded in both sheep and goats. All animals in this investigation were infected with multiple species to varying degrees. The mean burdens of adult nematodes were generally moderate in both sheep and goats and showed patterns of seasonal abundance that corresponded with the bi-modal annual rainfall pattern, with highest burdens around the middle of the rainy season. In both sheep and goats there were significant differences in the mean worm burdens and abundance of the different nematode species between the four geographic locations, with worm burdens in the Haramaya and Harar areas greater than those observed in the Dire Dawa and Jijiga locations. Similar seasonal variations were also observed in the prevalence of flukes. But there were no significant differences in the prevalence of each fluke species between the four locations. Overall, the results showed that Haemonchus, Trichostrongylus, Nematodirus, Oesophagostomum, Fasciola and Paramphistomum species were the most abundant helminth parasites of sheep and goats in eastern Ethiopia.  相似文献   

14.
Human and equine athletes are reported to have a high prevalence of gastric disease, and anecdotal evidence suggests a similar phenomenon applies to racing sled dogs. To investigate the prevalence of gastric disease in racing sled dogs, we conducted 2 gastroscopy studies on dogs competing in the annual Iditarod Sled Dog Race. A pilot study of dogs that were either dropped from the 2000 Iditarod Sled Dog Race because of illness or that finished the race indicated that, approximately 5 days after competing, 10 of 28 dogs (35%) had endoscopic evidence of gastric ulceration, erosion, or hemorrhage. The next year, an endoscopic study of 73 dogs participating in the 2001 Iditarod race was performed in order to evaluate a larger population of dogs. Data from 70 of these dogs could be used; 34 (48.5%) had ulceration, erosion, gastric hemorrhage, or some combination of these findings. When this group of 70 dogs was compared retrospectively to a control group of 87 dogs presented to the Texas A&M University (TAMU) Veterinary Medical Teaching Hospital, the Iditarod sled dogs had a significantly higher prevalence (P = .049) of gastric lesions. These findings suggest that, similar to athletes of other species, elite canine athletes have an increased prevalence of gastric disease compared to the canine population at large.  相似文献   

15.
The purpose of this study was to determine the frequency of different tumor types within a large cohort of cats with intracranial neoplasia and to attempt to correlate signalment, tumor size and location, and survival time for each tumor. Medical records of 160 cats with confirmed intracranial neoplasia evaluated between 1985 and 2001 were reviewed. Parameters evaluated included age, sex, breed, FeLV/FIV status, clinical signs, duration of signs, number of tumors, tumor location(s), imaging results, treatment, survival times, and histopathologic diagnosis. Most of the cats were older (11.3 +/- 3.8 years). Primary tumors accounted for 70.6% of cases. Metastasis and direct extension of secondary tumors accounted for only 5.6 and 3.8% of cases, respectively. Twelve cats (7.5%) had 2 or more discrete tumors of the same type, whereas 16 cats (10.0%) had 2 different types of intracranial tumors. The most common tumor types were meningioma (n = 93, 58.1%), lymphoma (n = 23, 14.4%), pituitary tumors (n = 14, 8.8%), and gliomas (n = 12, 7.5%). The most common neurological signs were altered consciousness (n = 42, 26.2%), circling (n = 36, 22.5%), and seizures (n = 36, 22.5%). Cats without specific neurological signs were common (n = 34, 21.2%). The tumor was considered an incidental finding in 30 (18.8%) cats. In addition to expected relationships (eg, meninges and meningioma, pituitary and pituitary tumors), we found that lesion location was predictive of tumor type with diffuse cerebral or brainstem involvement predictive of lymphoma and third ventricle involvement predictive of meningioma.  相似文献   

16.
Continuous rate infusion (CRI) of furosemide in humans is considered superior to intermittent administration (IA). This study examined whether furosemide CRI, compared with IA, would increase diuretic efficacy with decreased fluid and electrolyte fluctuations and activation of the renin-angiotensin-aldosterone system (RAAS) in the horse. Five mares were used in a crossover-design study. During a 24-hour period, each horse received a total of 3 mg/kg furosemide by either CRI (0.12 mg/kg/h preceded by a loading dose of 0.12 mg/kg IV) or IA (1 mg/kg IV q8h). There was not a statistically significant difference in urine volume over 24 hours between methods; however, urine volume was significantly greater after CRI compared with IA during the first 8 hours ([median 25th percentile, 75th percentile]: 9.6 L [8.9, 14.4] for CRI versus 5.9 L [5.3, 6.0] for IA). CRI produced a more uniform urine flow, decreased fluctuations in plasma volume, and suppressed renal concentrating ability throughout the infusion period. Potassium, Ca, and Cl excretion was greater during CRI than IA (1,133 mmol [1.110, 1,229] versus 764 mmol [709, 904], 102.7 mmol [96.0, 117.2] versus 73.3 mmol [65.0, 73.5], and 1,776 mmol [1,657, 2.378] versus 1,596 mmol [1,457, 1,767], respectively). Elimination half-lives of furosemide were 1.35 and 0.47 hours for CRI and IA, respectively. The area under the excretion rate curve was 1,285.7 and 184.2 mL x mg/mL for CRI and IA, respectively. Furosemide CRI (0.12 mg/kg/h) for 8 hours, preceded by a loading dose (0.12 mg/kg), is recommended when profound diuresis is needed acutely in horses.  相似文献   

17.
18.
The occurrence of ectoparasites in sheep flocks is frequently reported but seldom quantified. Sheep production used to be a predominantly family activity in the state of Sa~o Paulo (Brazil), but it began to become a commercial activity in the past decade. Thus, information about the ectoparasites existing in sheep flocks has become necessary. The present data were obtained by means of questionnaires sent to all sheep breeders belonging to the `Associaça~o Paulista de Criadores de Ovinos' (ASPACO; Sa~o Paulo State Association of Sheep Breeders). Response reliability was tested by means of random visits paid to 10.6% of the respondents. Most of the properties (89.5%) reported the presence of one or more ectoparasites. Screw-worm (Cochliomyia hominivorax) was the most frequent ectoparasite (72.5%), followed by bot fly larvae (Dermatobia hominis, 45.0%), ticks (Amblyomma cajennense) and Boophilus microplus, 31.3%) and finally lice (Damalinia ovis, 13.8%). Combined infestations also occurred, the most common one being screw-worm with bot fly larvae (36.0%) followed by bot fly larvae with ticks (13.9%), screw-worm with ticks (9.3%), bot fly larvae with lice (6.9%), and ticks with lice (5.0%). The most common triple combination was screw-worm, bot fly larvae and ticks (12.8%). Breeds raised for meat or wool were attacked by bot fly larvae and ticks more often than other breeds. Lice were only absent from animals of indigenous breeds. The relationships among these ectoparasites are discussed in terms of sheep breeds, flock size, seasonality and the ectoparasitic combinations on the host.  相似文献   

19.
The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.  相似文献   

20.
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号