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1.
为探讨DNA免疫防制传染性支气管炎(IB)的可能性,以克隆的Masschusete型类M41地方分离QD株S1基因为免疫原基因,引入终止密码后插入SV40启动子、增强子下游和SV40polyA信号上游,构建了S1基因DNA免疫表达质粒pSVQDS1。将大量扩增并经聚乙二醇纯化的pSVQDS1溶于PBS,以300μg/只的剂量肌肉注射免疫小鼠,于4周后采集分离免疫小鼠血清进行鸡胚病毒中和试验。结果表明,pSVQDS1能够诱导产生针对传染性支气管炎病毒(IBV)M41株的中和抗体,具有良好的免疫原性。  相似文献   

2.
用传染性支气管炎病毒(IBV)H120、H52、M41、ARK、澳大利亚T株、野外分离株RS、RY及鸡新城疫病毒(NDV)、鸡传染性法氏囊病毒(IBDV),鸡传染性喉气管炎病毒(ILTV)、减蛋综合症病毒(EDSV)等分别接种9日龄SPF鸡胚,37℃孵育48小时后,将其尿囊液离心,沉淀用PBS悬浮,涂片,用抗IBV单抗MC作一抗、进行间接荧光抗体检测。结果:H12O、H52、M41“ARK、T株及RS、RY均显阳性,而NDV、IBDV、ILTV、EDSV均为阴性。结果表明、用此法检测鸡胚尿囊液中IBV.具有快速、敏感、特异的优点。  相似文献   

3.
选择S1基因做为目的基因,运用RT-PCR技术分别扩增12株以非免疫鸡胚繁殖并通过超速离心浓缩的鸡胚尿囊液中的IBV,所用引物为IBVBeaudette株S1基因两侧的对应序列,跨幅为1.72kb。结果6株IBV毒株(SAIB3、F、D41、M41、H52、Holte)扩增出了长约1.7kb的S1基因片段,而另6株IBV毒株虽经4次RT-PCR重复后也显阴性。用HaeⅢ内切酶对6个毒株的RT-PCR产物进行酶切,结果显示M41、H52、D413个IBV毒株的S1基因包含两个HaeⅢ位点,F株与Holte株S1基因包含1个HaeⅢ位点,而SAIB3株S1基因则已丢失HaeⅢ位点。实验结果表明:运用RT-PCR技术检测IBV时,由于S1基因的核酸序列具有较高的变异率,因此S1基因不宜做为目的基因。  相似文献   

4.
用传染性支气管炎病毒H120、H52、M41、ARK、澳大利亚T株、野外分离株RS、RY及鸡新城疫病毒(NDV)、鸡传染性法氏囊病毒、鸡传染性喉气管炎病毒,减蛋综合症病毒等分别接种9日龄SPF鸡胚,37C孵育48小时后,将其尿囊液离心,沉淀用PBS悬浮,涂片,用抗IBV单抗MC作一抗,进行间接荧光抗体检测。结果:H120、H52、M41“ARK、T株及RS、RY均显阳性,而NDV、TBDV、ILT  相似文献   

5.
参照澳大利亚鸡传染性法氏囊病病毒( I B D V)00273 毒株基因组序列设计的 1 对引物,位于 V P2 高可变区两端,通过 R T P C R,扩增了 5 个 I B D V 山东分离株 V P2 基因的 607~1 080 位核苷酸片段(aa203~306)。 P C R 产物经纯化后,分别用 Dra Ⅰ、 Sac Ⅰ、 Ssp Ⅰ、 Pst Ⅰ和 Sau3 A I共 5 种限制性内切酶( R E)进行消化处理,结果 5 个分离株均为 Dra Ⅰ(- )、 Sac Ⅰ(- )和 Sau 3 A I(+ ), L C2 和 T A 株为 Ssp Ⅰ(+ ), L C1 和 J N 株为 Ssp Ⅰ(- ), L C1 和 L C2 株为 Pst Ⅰ(+ ), J N、 L L 和 T A 株为 Pst Ⅰ(- );5 个分离株的酶切图谱与美国变异株迥异,而 T A 株与日本超强毒株 9011 相类似。5 个分离株与现用疫苗株对比,至少在 V P2 可变区内存在着一定的差异,这可能是 I B D V 接种免疫鸡群仍然暴发 I B D 的原因之一。  相似文献   

6.
本研究对分离自沈阳地区的一株传染性支气管炎病毒(SY毒株)进行了生物学特性的研究,同时成功地对其免疫原S1基因进行了RT-PCR扩增、克隆与序列分析。 通过电镜观察、动物回归试验、血凝特性研究等试验验证分离自沈阳地区的SY毒株确实为一株传染性支气管炎病毒。气管环组织培养交叉中和试验结果表明,分离株SY株不同于参考毒株澳大利亚T、H52、M41,且不同于国内其它流行株HD、HB、XB、DB等,是一个新的变异株。 利用IBV S1基因特异性寡聚核苷酸引物,经RT-PCR扩增SY毒株的S1基因,得到预期的约1.7Kb片段;并将扩增所得cDNA插入克隆质粒pUC19的EcoRⅠ/BamHⅠ位点,在大肠杆菌DH5a中实现目的基因的克隆。经限制性核酸内切酶分析及PCR鉴定,证实为阳性重组质粒,利用末端双脱氧链终止法对其测序,得到S1基因全长1640bp,包括整个开放阅读框。通过序列分析软件DNASIS、PROSIS、MEGA等软件对S1基因核苷酸序列及推导的氨基酸序列进行分析,结果表明:分离株SY与7株参考株和国内流行株HD株相比,无论是核苷酸序列同源百分率还是氨基酸序列同源百分离都较低,均未达到80%,这就提示我们SY毒  相似文献   

7.
腺胃病变型鸡传染性支气管炎病毒变异株的比较研究   总被引:6,自引:2,他引:4  
腺胃病变型鸡传染性支气管炎病毒(IBV)变异株H95提纯样品经SDS-PAGE和Westernbloting,证明H95株单抗DE7和M41株单抗6DH8均能识别H95株54000蛋白多肽。采用单抗、多抗介导间接ELISA试验表明,H95毒株能与抗IBVM41N蛋白单抗6DH8和IBVM蛋白单抗MC发生反应,也能与抗M41、Gray、Holte、T、H52、N115和分离株C9001、DLZ9111的多抗血清反应,同时抗H95株的4株单抗、多抗血清也能与上述毒株反应。卵磷脂酶C处理的H95株的血凝活性能被相应的H95株的单克隆抗体和高免血清所抑制,也能被M41单抗和高免血清所抑制。通过RT-PCR获得了H95毒株的免疫原基因S1,经Southernbloting和IBV的S探针检测呈阳性。  相似文献   

8.
牛传染性鼻气管炎病毒TK基因缺失株的构建   总被引:8,自引:1,他引:7  
将含有牛传染性鼻气管炎病毒( I B R V) T K 基因的片段克隆于p Bluescript S K 中, 再用 Bg1 Ⅱ和 Sac I切去 T K 基因中347bp 的片段, 获得了含缺失 T K 基因的重组质粒, 然后插入 Lac Z 报告基因, 构建成转移载体。以脂质体转染法将此转移载体与 I B R V Bartha 株在牛肾细胞( M D B K) 上共转染, 经过蓝色蚀斑筛选和蚀斑克隆纯化, 获得了能稳定遗传的重组 I B R V。经 P C R 和 Southern 印迹杂交鉴定, 证实该重组病毒为 T K 基因缺失突变株。  相似文献   

9.
禽传染性支气管炎病毒免疫原基因组的RT—PCR研究   总被引:9,自引:0,他引:9  
本文报道了用聚合酶链反应获取IBV M41株和广东肾变栋D41免疫基因的研究情况,所用引物为IBV Beaudette株S1基因两侧的对应序列,跨幅为1.7kb;DNA模板为IBVM41和D41株经病毒RNA提取后反转录而得;2株病毒所获的PCR产物与预期一致。  相似文献   

10.
青海细毛羊生化遗传多态性的研究   总被引:7,自引:0,他引:7  
采用聚丙烯酰胺凝胶电泳和火焰光度法对509只青海细毛羊细血液和乳汁中HB,EP-1,EP-2,KE,TF,AMY1,AMY2,ES,ALP,RBC-LDH,Hpα-LA和β-LG总共13个位点的多态性进行了研究。结果为:(1)在青海细毛羊的HB,KE,TF,AMY2,ES,ALP,RBC-LDH1,Hp,β-LG总共9个位点上发现多态性,多态性位点的比例为69.23%;(2)每个位点实有的平均等位  相似文献   

11.
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   

12.
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   

13.
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   

14.
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   

15.
Genetic variations in chromosome Y are enabling researchers to identify paternal lineages, which are informative for introgressions and migrations. In this study, the male‐specific region markers, sex‐determining region‐Y (SRY), amelogenin (AMELY) and zinc finger (ZFY) were analysed in seven Turkish native goat breeds, Angora, Kilis, Hair, Honaml?, Norduz, Gürcü and Abaza. A SNP in the ZFY gene defined a new haplotype Y2C. All domestic haplogroups originate from Capra aegagrus, while the finding of Y1A, Y1B, Y2A and Y2C in 32, 4, 126 and 2 Turkish domestic goats, respectively, appears to indicate a predomestic origin of the major haplotypes. The occurrence of four haplotypes in the Hair goat and, in contrast, a frequency of 96% of Y1A in the Kilis breed illustrate that Y‐chromosomal variants have a more breed‐dependent distribution than mitochondrial or autosomal DNA. This probably reflects male founder effects, but a role in adaptation cannot be excluded.  相似文献   

16.
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   

17.
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   

18.
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   

19.
20.
An experiment was conducted to study disappearance of C14 to C18 fatty acids, lag times and biohydrogenation (BH) rates of C18 fatty acids of ground soya bean and canola seeds in situ. Three ruminally fistulated Dallagh sheep were used to determine ruminal BH of unsaturated fatty acids (UFAs). Differences in the disappearance of fatty acids through the bags and lag times were observed between the oilseeds. We saw that the longer the incubation time of the oilseeds in the rumen, the lower the content of C18:2 and C18:3. Significantly higher lag times for both C18:2 and C18:3 were observed in ground canola compared to ground soya bean. BH rates of C18:2 and C18:3 fatty acids in soya bean were three times higher than those of canola. These results suggest that the fatty acid profile of fat source can affect the BH of UFAs by rumen micro‐organisms. So that UFAs of canola had higher ability to escape from ruminal BH. It seems that fatty acid profile of ruminant products is more affected by canola seed compared to soya bean seed.  相似文献   

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