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1.
以传染性法氏囊病病毒(IBDV)浙江分离株(ZJ2000)基因组RNA为模板,采用Long-accurate RT-PCR(LA-PCR)一步法扩增并克隆了IBDV ZJ2000株基因组A节段全长cDNA。序列测定结果表明,克隆的A节段全长共3259个核苷酸,包括5'、3'端的非编码区(NCRs)和2个部分重叠的开放阅读框(ORF1和ORF2),与参比的血清1型毒株核苷酸序列的同源性高达95.2%-99.2%。二级结构预测表明,在5'-NCRs存在1个大型的茎环发夹结构。ORF2编码145个氨基酸的VP5,与参比毒株的同源性高达98.6%-100%。ORF1编码1012个氨基酸的VP2/VP4/VP3,在氨基酸水平上VP2、VP3、VP4与参比毒株的同源性分别达到94.9%-98.8、96.1%-98.5%、97.1%-99.2%。ZJ2000共有14-38氨基酸的替代,其中特有氨基酸6个,变异大多数集中在VP2高变区,突变率达2.9%-11%。第2个小亲水区内280位氨基酸由S替代了N、290位M替代了L。这2个突变可能与IBDV的抗原性有关。VP2-VP4剪切位点附近511和540位氨基酸的变异可能使ZJ2000株的毒力增强。分子系统进化树分析表明,ZJ2000与欧洲Cu-1株、P2株、CEF94株和中国Harbin株的有关最近,而与欧洲、香港、日本的超强毒株和美国的变异株相对较远。  相似文献   

2.
根据国外已发表的鸡传染性支气管炎病毒(IBV)S1基因序列,设计了两对引物并以RT-PCR特异性扩增出嗜肾型IBV W93疫苗株的S1基因,基因产物大小为1.62kb,与设计相符,对其进行序列测定后,与标准毒株H52、H120、M41、BEAU株的S1基因进行同源性比较,结果表明,W93株与H52、H120、M41和BEAU株的核苷酸序列的同源性分别为96.8%、97.1%、96.9%和96.8%,由此可以看出嗜肾型IBV W93疫苗株与标准毒株在S1基因上具有高度的同源性。  相似文献   

3.
鸡传染性支气管炎病毒中国地方分离株S1基因5''端的变异   总被引:3,自引:0,他引:3  
对自海南海省、广西省发生鸡传染性支气管炎(IB)鸡群分离的4株IBV分离株(HaN-1/95、HaN-2/95、GX-1/98、GX-2/98)的主要免疫原纤突蛋白S1基因经RT-PCR扩增其5'端的1.2kb的目的片段,将其插入载体pMD 18-Tk ,在大肠杆菌中实现目的的基因的克隆。对克隆的目的基因经限制性酶切分析及PCR鉴定后,以双脱氧链终止法测定其核苷酸序列,并与GenBank中的参考毒株(H120、SD-1/97和Holte)相应序列作比较,分析其同源性。结果表明,HaN-1/95、HaN-2/95、GX-1/98及SD-1/97与疫苗株H120的核苷酸序列同源性分别为99.5%、99.2%、97.9%和99.5%,其推导氨基酸序列同源性分别为99.1%、98.9%、96.9%和99.2%。GX-2/98与Holte株的核苷酸序列同源率为99.0%,其推导的氨基酸序列同源性为98.6%,而与其他中国分离株的核苷酸序列的同源性仅为70%左右,氨基酸序列的同源性仅为68%左右。  相似文献   

4.
5.
对鸽源冠状病毒PSH株纤突蛋白基因进行了RT—PCR扩增、克隆和测序。结果表明,PSH株S基因由3504个核苷酸组成,编码1条由1167个氨基酸残基组成的多肽。S基因编码产物裂解后形成的S1和S2亚单位分别由541和626个氨基酸残基组成。PSH株S蛋白切割识别位点为精氨酸-精氨酸-苯丙氨酸-精氨酸-精氨酸(RRFRR),与多数鸡传染性支气管炎病毒(IBV)切割识别位点相似(RRF/SRR)。与GenBank中其他冠状病毒毒株S基因的推导氨基酸序列相比较,PSH株与IBV参考毒株的同源性为79.3%~99.6%,而与其他冠状病毒包括火鸡蓝冠病病毒和SARS病毒的同源性均小于37.8%。表明,鸽源冠状病毒PSH株属于第3抗原群冠状病毒,且与IBV亲缘关系较近。  相似文献   

6.
参照已发表的鸡传染性支气管炎病毒(infectious bronchitis virus,IBV)基因序列,设计合成了1对引物,经RT—PCR扩增得到了IBV国内分离株D971约1.3kb的片段,其中包含3a、3b、E及M蛋白基因的完整0RF,并将该片段进行了克隆和序列测定。与国内外部分IBV毒株序列比较表明:D9713a基因与CU—T2、D1466、DE072、M41、GA/99和UK/66的3a基因核苷酸序列同源性在83%~89%之间,氨基酸序列同源性为77%~91%,其中与CU—T2毒株的同源性最低(分别为83%和77%);3b基因与上述毒株的对应核苷酸序列同源性在85%~95%之间,氨基酸序列同源性为72%~96%,与CU—T2的同源性最高(分别为95%和96%);E蛋白基因与CU—T2、D1466、DE072、M41、Ark99、H52、Gray、GA/99和UK/66的核苷酸序列同源性在81%~86%之间,氨基酸序列同源性为74%~80%。不同毒株推导的E蛋白C末端表现不同特征,其中D971E蛋白C末端比CU—T2的时应区多9个氨基酸残基,而比本试验选用的其他参考毒株的相应区域短5~7个氨基酸残基。M基因与Cu—T2、DE072、Gray、M41、H120、H52、SD/97/02、LX4及D41的M基因核苷酸序列同源性在86%~91%之间,氨基酸序列同源性为85%~95%,其中与SD/97/02的同源性最高(分别为91%和95%)。对M蛋白的3个功能区进行序列比较发现,其N端第1~19位与SD/97/02的同源性最高,达90%,与CU—T2的同源性为80%,而与其他毒株同源性均低于61%;N端第20~100位含有3个跨膜蔬水区,D971与其他参考毒株比较,该部位第20~67位相对保守,其中与SD/97/02的同源性为100%,与其他毒株的同源性在97%以下;在推测的与核衣壳连接的C端,D971与SD/97/02在第101~182位同源性为100%,与疫苗株H52和H120及国内分离株LX4和D41的同源性为97%,而与本试验所选的国外参考毒株同源性在96%以下;在C端第184~219位D971与H52的同源性为100%,与H120和SD/97/02的同源性为97%,与其他毒株的同源性也在96%以下,而且在C末端第220~222及224位与上述国内外参考毒株均不同,国内外各毒株在该4个位点氨基酸为丙氨酸、苏氨酸、甘氨酸和丝氨酸,而D971M蛋白的该区域氨基酸突变为丝氨酸、丙氨酸、缬氨酸和甘氨酸,该突变是否与D971的生物学特性尤其是组织嗜性的变化相关还有待于进一步研究。以上结果表明,D971与SD/97/02亲缘关系较其他毒株为近,但其遗传衍化又与国内使用的疫苗株H52和H120具有一定的联系。从其mRNA3和mRNA4cDNA的序列分析来看,D971又具有独特的分子特征。  相似文献   

7.
根据国外已发表的鸡传染性支气管炎病毒(IBV)S1基因序列设计了一对引物,通过:RT-PCR特异性扩增出IBV H120疫苗株的S1基因,产物大小为1.62kb,与设计相符。同源性比较结果显示,H120株与H52、M41和BEAU株的S1基因核苷酸序列的同源性分别为97.1%、96.9%和96.8%,表明IBV H120疫苗株与标准毒株的S1基因具有高度的同源性。  相似文献   

8.
采用RT-PCR方法扩增了鸡传染性支气管炎病毒D971毒株预期的1636bp的S1全基因DNA片段;以Sanger‘s双脱氧末端终止法测定出其核苷酸序列,推导出了氨基酸序列,并用有关软件构建了S1基因进化树。结果表明,IBV-D971毒株与国内外AF140352(山东农业大学)、AF154841(山东农业大学)、AF193432(青岛动物检疫所)、AY043312(中国农业大学)、AF397528(四川农业大学)、AY043221(浙江农业大学)、AF352312(浙江农业大学)、U29522(澳大利亚)和M21883(英国)毒株的核苷酸同源性分别为99%、99%、95%、94%、87%、86%、88%、82%和80%。氨基酸序列同源性依次分别为93%、93%、87%、83%、83%、82%和80%。  相似文献   

9.
本研究选取有代表意义的9株鸡传染性支气管炎病毒(IBV)国内分离株.利用所设计的一对特异引物,通过RT—PCR的方法成功地扩增了膜蛋白基因即M基因全长片段.通过克隆、序列测定获得了9个分离毒株M基因的全长核苷酸序列。并将各IBV国内分离株与GenBank中注册的一些毒株的M基因核苷酸序列及推导的氨基酸序列进行比较、系统进化关系分析发现:IBV中国地方分离毒株大部分属于Mass型;不同毒株间存着重组、缺失、插入及点突变等变异,从ATG至第150bp区段的核苷酸序列变异频率最高;IBV中国地方分离株M基因的变异属于高频率的同义突变;国内的IBV分离株分为两个基因群,1群分离毒株与GenBank中Mass型的毒株亲缘关系较近.毒株间核苷酸序列同源性为95.7%~100%,其相应的氨基酸序列同源性为98.2%~100%;Ⅱ群分离毒株与众多参考毒株的亲缘关系都比较远,毒株间彼此核苷酸序列同源性为89.7%~91.9%,氮基酸序列同源性为92.0-96.0%。  相似文献   

10.
4株猪瘟流行野毒株病毒结构蛋白E2基因的核苷酸序列分析   总被引:6,自引:0,他引:6  
应用RT-PCR和nPCR扩增由4株甘肃省近期(1997-1998)猪瘟流行野毒株的E2基因,将其克隆到pGEM-TEasy载体上,经转化、筛选、鉴定后,测定核苷核序列,4株流行毒株的E2基因核苷酸序列同源性为89.2%-99.7%,相应的氨基酸序列同源性为93.8%-99.0%,这4株流行毒株;与C-株(疫苗种毒)E2基因的核苷酸序列同源性为82.2%-84.3%,相应的氨基酸序列的同源性为87.9%-90.2%,表明近期猪猪瘟流行毒株与C-株的gp55蛋白之间存在一定的差异。  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.
15.
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   

16.
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   

17.
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   

18.
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   

19.
The objective of this report was to characterize 20-year changes in proportion of calcium oxalate (CaOx) calculi and struvite calculi in dogs, and associations with breed, age, and sex. In this retrospective study, results of analysis of urinary calculi from dogs were reviewed for specimens received between July 1, 1981, and December 31, 2001. Breed, sex, age, year of submission of the specimen, and mineral type(s) were analyzed statistically. CaOx or Struvite or both were contained in 18,966 of 20,884 (91%) specimens. For both sexes, a 20-year statistically significant increase was observed in the proportion of calculus specimens that contained CaOx. The increase in this proportion was greater in females (1% to 31%) than in males (18% to 82%). From 1998 to 2001, when proportions may have plateaued, the odds of specimens containing CaOx were markedly higher in 18 breeds, markedly lower in 5 breeds, and not significantly different in 13 breeds compared with crossbreds. For both sexes, a 20-year statistically significant decrease was observed in the proportion of calculus specimens that contained struvite. This decrease in proportion was greater for males (79-16%) than for females (97-68%). From 1998 to 2001, when proportions plateaued, the odds of calculi containing struvite were markedly lower in 20 breeds, markedly higher in 1 breed, and not significantly different in 15 breeds when compared with crossbreds. Breed, age, and sex were associated statistically with CaOx or struvite urolithiasis. In conclusion, there appears to have been a long-term increase in the proportion of specimens of canine urinary calculi that contain CaOx as well as a long-term decrease in the proportion of specimens of calculi that contain struvite for both male and female dogs. The rate of change appeared to begin leveling off in the period 1998 to 2001. The recent proportion of dogs with either CaOx- or struvite-associated urolithiasis may depend on breed, age, and sex, and on interactions among these 3 factors.  相似文献   

20.
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   

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