共查询到20条相似文献，搜索用时 599 毫秒
1.
《中国兽医学报》2017,(6)
本研究建立了绵羊梨型虫的检测方法,并对退火温度和循环次数进行梯度优化。选取条带最亮且无杂带的温度和循环次数作为PCR检测方法的优化结果,将新建的PCR方法与传统的吉姆萨染色法结果进行对比,将阳性的PCR产物进行测序。结果表明:PCR反应的最佳体系为94℃预变性5 min;94℃变性1 min,56℃退火90s,72℃延伸1min,共循环39次;72℃延伸10min,4℃保存;在50份临床样品的检测中,PCR法检出的阳性样品有13份,阳性率为26%(13/50),吉姆萨染色发检出的阳性样品有8份,阳性率为16%(8/50),由PCR方法检测出的阳性样品与吉姆萨染色发检出的阳性样品符合率达100%。阳性PCR产物的测序结果的同源性分别为100%和98%。本研究证实PCR法的灵敏度和准确性高于吉姆萨氏染色镜检法。 相似文献
2.
为了明确PCR产物直接测序与克隆后测序结果的差异,验证PCR产物直接测序的“准确性”,将从两个规模化猪场采集的组织样品（A、B）PRRSV ORF5 PCR阳性扩增产物胶回收后连接至pMD19-T载体,分别取15个阳性亚克隆单菌落,同原始PCR产物进行测序,对两个样品各获得的16条序列进行比对分析。结果显示：A、B两组样品16条序列间的核苷酸同源性分别为84.2%~100%、87.4%~100%,PCR产物与其15个亚克隆测序获得序列的核苷酸同源性分别为87.7%~97.3%、87.9%~100%。遗传演化分析方面：样品A序列与其亚克隆序列被划分在谱系1、5、8,占比分别为6.25%、87.50%、6.25%,与其12个亚克隆序列同属于谱系5;样品B与其亚克隆序列被划分在谱系5、8,占比分别为68.75%、31.25%,与其10个亚克隆序列同属于谱系5。结果表明,同一样品中PRRSV ORF5序列具有多样性,不同样品的PRRSV ORF5序列丰富度不同。结果提示,在生产实际中,通过PCR对PRRSV ORF5产物进行直接测序,对于了解场内流行毒株具有一定的指示意义。 相似文献
3.
2018年以来,某进口品种肉种鸡发生疑似禽白血病病例,流行范围广,危害严重,为确定该次疫情的病原,本研究从辽宁、山东等地发病鸡场采集55份疑似禽白血病病料样品。病理组织切片观察显示,病料样品肝脏和脾脏均有髓细胞样肿瘤细胞增生。针对禽白血病病毒(ALV)、鸡马立克氏病病毒(MDV)和网状内皮组织增生症病毒(REV)的特异性PCR检测结果显示,55份病料样品中ALV阳性样品12份(21.8%),MDV阳性样品1份(1.82%),REV均为阴性。将12份ALV阳性病料样品经处理后接种DF-1细胞进行病毒分离,ELISA检测结果显示有8份ALV群特异性抗原阳性。ALV多重PCR检测结果表明,8个分离株均为J亚群ALV(ALV-J),测序结果显示,8个分离株之间核苷酸序列同源性为99.5%~100%,与A、B、C、D、E亚群ALV的同源性仅为56.5%~58.8%,与ALV-J的同源性为90.7%~96.6%,进一步表明所有分离株均为ALV-J。上述结果表明,引起该次进口白羽父母代肉种鸡发生禽白血病的病原主要以ALV-J为主,该研究结果为疫情的溯源和有效防控奠定了基础。 相似文献
4.
为全面了解吉林省某规模化牛场牛病毒性腹泻(BVD)的流行情况,试验采集临床血清样品157份,粪便样品18份,肝脏、精液等组织样品17份,应用BVDV抗体检测试剂盒进行血清抗体检测,利用BVDV1型引物,采用纳米PCR方法对血清及临床样品进行BVDV抗原检测,对抗原阳性样品进行测序分析;将抗原阳性样品经研磨、稀释后用0.22μm滤膜过滤除菌,接入牛肾细胞(MDBK)进行病毒分离培养,盲传3代后再次进行抗原检测;采用免疫荧光技术检测病毒对MDBK细胞的侵染作用;利用Mega软件绘制系统进化树并进行同源性比对分析。结果显示,该牛场临床血清BVDV抗体阳性率为77.1%,血清抗原阳性率为12.1%,临床粪便等样品抗原阳性率为74.3%;病料接入细胞未观察到细胞病变,分离毒株PCR产物测序分析结果与样品抗原检测结果一致;免疫荧光检测结果显示,分离株毒液正常吸附于MDBK细胞中,有明显荧光反应;抗原测序分析显示,该牛场BVD主要流行毒株与BVDV JL-1株同源性高达99.0%,且均为BVDV 1型毒株。本研究对该牛场BVDV流行情况进行了全面调查,为开展净化工作奠定了基础。 相似文献
5.
猪细小病毒2型（porcine parvovirus type 2,PPV2）是2001年首次在缅甸发现的新型病毒。为深入了解四川省绵阳市PPV2感染及遗传变异情况,使用普通PCR对采自4个地区10个规模化猪场的296份临床病猪血清样本进行了核酸检测,并对其中1份阳性样品进行了全基因组测序和遗传变异分析。结果显示:PPV2检出率为1.69%（5/296）;阳性样品（PPV2-11-MY）全基因组长度为5 506 bp,与12个PPV2参考毒株的核苷酸同源性为93.96%～99.30%,氨基酸同源性为52.50%～73.80%;存在3处氨基酸高变区域,分别位于91～218 aa、513～626 aa和776～1 065 aa;该毒株同源性最接近我国江苏省毒株MW853949.1。本研究不仅丰富了PPV2流行病学数据,为后续疫苗研究提供了理论依据,同时也为进一步研究PPV2致病机理及毒力奠定了基础。 相似文献
6.
7.
沙门菌和志贺菌二重PCR检测方法的建立及应用 总被引：3，自引：2，他引：1
根据GenBank提供的沙门菌invA基因序列和志贺菌ipaH基因序列设计引物,建立了二重PCR方法,在同一反应体系同时检测两种致病菌核酸,经二重PCR方法扩增后,在同一泳道同时检出沙门菌284 bp和志贺菌611 bp特异性扩增条带,而普通大肠埃希菌、金黄色葡萄球菌、变形杆菌、阪畸大肠埃希菌、绿脓杆菌、蜡样芽胞杆菌、马链球菌、产单核细胞李斯特菌、鹅大肠埃希菌、猪水肿病大肠埃希菌均未出现这两种条带.对PCR产物进行测序,测序结果与已发表的基因核苷酸序列比较,沙门菌同源性为93%～100%,志贺菌同源性为98%～100%.应用建立的沙门菌和志贺菌二重PCR方法对广西6个试验猴养殖场1 665份猴粪便样品进行检测,检出沙门菌阳性25份,志贺菌阳性90份,阳性检出率分别为1.5%和5.4%.表明建立的沙门菌和志贺菌二重PCR检测方法特异性强、敏感性高,适用于临床粪便样品的快速检测. 相似文献
8.
根据GenBank中已发表的猪细小病毒(porcine parvovirus,PPV)、猪伪狂犬病病毒(pseudorabies virus,PRV)和猪圆环病毒2型 (porcine circovirus type 2,PCV2)基因序列,对各病毒基因区进行同源性分析,确定PPV 的VP2、PRV的 gD、和PCV2的ORF2基因为各病毒的诊断靶序列,设计特异性引物,在建立各病毒单项PCR技术的基础上,优化多重PCR反应条件,建立了3种病毒的多重PCR技术,可同时扩增PPV 313 bp、 PRV 217 bp和PCV2 447 bp的特异性片段。用多重PCR技术与单项PCR技术对比检测试验证明两者的符合率为100％,表明建立的多重PCR检测方法,具有特异、快速、准确的特点,可同时鉴别诊断这3种病毒。从10个发病猪场和门诊病例的病猪采集的211份样品,用建立的多重PCR检测方法,检出PPV阳性42份,阳性率为19.91%;PRV阳性26份,阳性率为12.32%;PCV2阳性56份,阳性率为26.54%;2种以上病毒混合感染25份,混合感染阳性率为11.85%。检测结果表明,山西省猪群已感染这3种疫病。 相似文献
9.
10.
为全面了解吉林省某规模化牛场牛病毒性腹泻(BVD)的流行情况,试验采集临床血清样品157份,粪便样品18份,肝脏、精液等组织样品17份,应用BVDV抗体检测试剂盒进行血清抗体检测,利用BVDV1型引物,采用纳米PCR方法对血清及临床样品进行BVDV抗原检测,对抗原阳性样品进行测序分析;将抗原阳性样品经研磨、稀释后用0.22μm滤膜过滤除菌,接入牛肾细胞(MDBK)进行病毒分离培养,盲传3代后再次进行抗原检测;采用免疫荧光技术检测病毒对MDBK细胞的侵染作用;利用Mega软件绘制系统进化树并进行同源性比对分析。结果显示,该牛场临床血清BVDV抗体阳性率为77.1%,血清抗原阳性率为12.1%,临床粪便等样品抗原阳性率为74.3%;病料接入细胞未观察到细胞病变,分离毒株PCR产物测序分析结果与样品抗原检测结果一致;免疫荧光检测结果显示,分离株毒液正常吸附于MDBK细胞中,有明显荧光反应;抗原测序分析显示,该牛场BVD主要流行毒株与BVDV JL-1株同源性高达99.0%,且均为BVDV 1型毒株。本研究对该牛场BVDV流行情况进行了全面调查,为开展净化工作奠定了基础。 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.
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
14.
Tropical Animal Health and Production - 相似文献
15.
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
16.
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
17.
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
18.
B. Cinar Kul N. Bilgen J.A. Lenstra O. Korkmaz Agaoglu B. Akyuz O. Ertugrul 《Zeitschrift für Tierzüchtung und Züchtungsbiologie》2015,132(6):449-453
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
19.
Ling GV Thurmond MC Choi YK Franti CE Ruby AL Johnson DL 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2003,17(6):817-823
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.
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