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1.
鸡传染性法氏囊病病毒野毒株VP2基因高可变区酶切位点分析 总被引:5,自引:1,他引:4
参照澳大利亚鸡传染性法氏囊病病毒( I B D V)00273 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 C2 和 T A 株为 Ssp Ⅰ(+ ), L C1 和 J N 株为 Ssp Ⅰ(- ), L C1 和 L C2 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 9011 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
2.
传染性支气管炎病毒S1基因DNA免疫质粒的构建及其免疫原性 总被引:7,自引:0,他引:7
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
3.
用单抗介导的荧光抗体技术检测鸡尿囊细胞内传染性支气管炎病毒的研究 总被引:2,自引:0,他引: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
4.
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
5.
用单抗介导的荧光抗体技术检测鸡尿囊细胞内传染性支气管炎病毒的研究 总被引:3,自引:0,他引:3
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
6.
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:4,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
7.
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
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.
Rassnick KM Gieger TL Williams LE Ruslander DM Northrup NC Kristal O Myers NC Moore AS 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2001,15(3):196-199
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.
Furosemide continuous rate infusion in the horse: evaluation of enhanced efficacy and reduced side effects 总被引:1,自引:0,他引:1
Johansson AM Gardner SY Levine JF Papich MG LaFevers DH Fuquay LR Reagan VH Atkins CE 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2003,17(6):887-895
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
13.
Davis MS Willard MD Nelson SL Mandsager RE McKiernan BS Mansell JK Lehenbauer TW 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2003,17(3):311-314
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
14.
Tropical Animal Health and Production - 相似文献
15.
Waruiru RM Thamsborg SM Nansen P Kyvsgaard NC Bogh HO Munyua WK Gathuma JM 《Tropical animal health and production》2001,33(3):173-187
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
16.
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
17.
H Hayashi T Yoshimura J Y Chen 《Comparative immunology, microbiology and infectious diseases》1985,8(2):73-87
Our particular attention in this article was given to natural mediators for macrophages isolated from the sites of tissue injury. A number of chemotactic factors, which may satisfy many criteria making them acceptable as inflammatory leucocyte chemotactic factors, has been separated. Among them, our laboratory has isolated three macrophage (monocyte) chemotactic factors (MCF-a, -b and -c). Their purification, characterization and functional specificity are discussed. 相似文献
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Evaluation of biohydrogenation rate of canola vs. soya bean seeds as unsaturated fatty acids sources for ruminants in situ
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S. Pashaei T. Ghoorchi A. Yamchi 《Journal of animal physiology and animal nutrition》2016,100(2):211-216
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
20.
Serum biochemical values and the wet weight to dry weight ratios of tissues were determined in yaks in Shandan county of Gansu province. The liver, kidney, heart and muscle contents of seven elements in yaks were also analysed. Most serum biochemical values were similar to those of cattle, camels and sheep, but the calcium concentration was considerably above the normal range for other ruminants. The liver contained the highest concentrations of copper, manganese and cobalt and the kidney of selenium, iron and calcium.Abbreviations Alb
albumin
- ALT
alanine aminotransferase
- AST
aspartate aminotransferase
- BUN
blood urea nitrogen
- Chol
cholesterol
- Glob
globulin
- -GT
-glutamyl transpeptidase
- IP
inorganic phosphorus
- TP
total protein 相似文献