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1.
为进一步探讨旋毛虫P53基因原核表达重组蛋白的免疫保护作用,本实验采用纯化的重组蛋白作为抗原免疫小鼠,免疫剂量为50μg/100μL,每隔10d免疫1次,共3次。末次免疫后10d,每只小鼠以200条旋毛虫肌幼虫攻虫。分别检查感染后7d小鼠肠道内成虫数量、体外培养雌虫所产新生蚴数量以及感染后35d肌肉荷虫量。结果显示:小鼠肠道内成虫、新生蚴和肌幼虫的减虫率分别为61.9%、53.45%和71.48%,表明旋毛虫表达P53基因的重组蛋白对小鼠产生了较好的免疫保护效果。  相似文献   

2.
各旋毛虫隔离种某些生物学特性研究   总被引:3,自引:0,他引:3  
通过对猪、犬旋毛虫和国际标准隔离种:旋毛形线虫(Trichinella spiralis)和本地毛形线虫(Trichinella nativa)的研究发现,猪旋毛虫和旋毛形线虫在小鼠隔肌中出现保姆细胞的时间比较早,分别于感染第16天和18天出现,第38天和36天所有幼虫都已形成保姆细胞,而犬旋毛虫和本地毛形线虫出现保姆细胞的时间较晚,于感染第20天和22天出现,第32天完全形成。猪旋毛虫和旋毛形线虫雌虫体外培养24小时平均产新生幼虫数分别为66.0和76.2,而犬旋毛虫和本地毛形线虫分别是28.8和22.0,前二者在雌虫体外产新生幼虫能力上明显高于后二者。研究结果表明,黑龙江猪旋毛虫为旋毛形线虫,犬旋毛虫为本地毛形线虫。  相似文献   

3.
为了获得本地毛形线虫(Trichinella nativa)49 ku ES蛋白的结构基因,用Trizol提取T.nativa肌幼虫的总RNA,用RT-PCR方法扩增出了编码T.nativa49 ku排泄分泌蛋白的结构基因。将PCR产物与T载体连接并经过大肠埃希菌增殖后送检测序。序列测定表明,目的基因TNPG长度为951bp,核苷酸序列同已发表的T.spiralis相应的序列P49同源性为98.63%,所推导的氨基酸序列同源性为99.05%。为T.nativa49 ku ES蛋白结构基因的进一步研究打下基础。  相似文献   

4.
旋毛虫感染小鼠对p46 000重组抗原的抗体应答   总被引:1,自引:0,他引:1  
分别以旋毛虫肌幼虫ES抗原和p46000重组蛋白作为抗原,对小鼠人工感染旋毛虫后的抗体应答进行了ELISA检测。结果表明,以肌幼虫200条/只经口感染小鼠后,肌幼虫ES抗原在感染后9d可检出抗体,并于感染后35~42d达到最高水平;应用重组抗原检测时,感染后10d可检出抗体,抗体水平略低于用ES抗原,但是其消长规律基本一致.而且与阴性血清相比差异明显;抗体在117d后仍维持于较高水平。  相似文献   

5.
根据BAC-TO-BAC杆状病毒表达系统的供体质粒,设计表达引物,从克隆质粒上扩增本地毛形线虫(Trichinella nativa)49kuES结构蛋白基因TNPG,酶切后与供体质粒连接,在大肠杆菌中克隆,将克隆有外源基因的供体质粒转化DH10Bac感受态菌,该菌含有杆状病毒穿梭载体,供体质粒中的外源基因在辅助质粒编码的转座酶作用下,通过转座而插入杆状病毒基因组中,筛选含有重组质粒的BacmidDNA,用脂质体介导法转染昆虫细胞获得重组病毒。SDS-PAGE电泳显示表达蛋白为36ku的融合蛋白,Westernblot检测表明表达的融合蛋白能够被小鼠旋毛虫的阳性血清识别。实验结果表明,本地毛形线虫的ES融合蛋白的制备将在诊断方法的建立和免疫研究方面具有很好的应用前景。  相似文献   

6.
为研究日本血吸虫凋亡蛋白抑制因子(inhibitor of apoptosis protein of Schistosoma japonicum,SjIAP)重组蛋白诱导BALB/c小鼠的免疫保护效果,利用PCR技术扩增SjIAP基因,构建重组表达质粒pET-28a(+)-SjIAP,诱导表达重组SjIAP蛋白,并利用重组蛋白制备兔源多克隆抗体血清。然后,选用SjIAP重组蛋白免疫BALB/c小鼠,利用ELISA检测免疫小鼠血清中的特异性抗体水平,以及免疫小鼠脾脏淋巴细胞在SjIAP重组蛋白刺激后产生的细胞因子水平。强化免疫后,将小鼠进行血吸虫尾蚴攻虫试验,感染38 d,进行剖杀,计算虫体减虫率及肝脏减卵率。Western blot结果表明本研究制备的兔源多抗血清能特异性识别SjIAP重组蛋白。ELISA检测表明免疫SjIAP重组蛋白可诱导较高水平的IgG及IgG亚型(IgG1、IgG2a、IgG2b、IgG3)抗体和IFN-γ、IL-2及IL-4细胞因子。动物试验表明,免疫SjIAP重组蛋白的小鼠与PBS组相比分别获得了31.5%的减虫率和37.2%的肝脏减卵率。免疫SjIAP重组蛋白能诱导小鼠获得一定减虫和减卵保护效果,提示血吸虫凋亡蛋白抑制因子可作为抗血吸虫病的疫苗候选分子。  相似文献   

7.
《中国兽医学报》2015,(8):1284-1289
根据GenBank上旋毛虫Kazal型丝氨酸蛋白酶抑制剂(KaSPI)基因编码序列设计特异性引物,进行RT-PCR扩增,将所获PCR产物克隆至pEASY-T1载体后转入克隆感受态Tans5α,经过PCR和EcoRⅠ、XhoⅠ双酶切鉴定后进行测序。将所获目的基因与载体pET-30a(+)相连接,然后将鉴定正确的重组表达质粒pET-TsKaSPI转化到大肠杆菌BL21(DE3)中,利用IPTG诱导蛋白表达。SDS-PAGE电泳分析所得融合蛋白大小约为38 000,与预测蛋白理论值大小相符,主要以包涵体形式存在。对纯化后的重组蛋白进行Western blot鉴定,结果显示该蛋白可以被感染旋毛虫小鼠阳性血清所识别,具有良好的抗原性。将纯化的重组蛋白经腹腔注射到小鼠体内检测其对小鼠的免疫保护性,结果表明,旋毛虫肌幼虫减虫率为38.3%。通过间接ELISA法检测血清抗体水平,结果显示重组蛋白免疫小鼠血清的抗重组蛋白抗体IgG总体水平显著高于感染对照组和佐剂组。  相似文献   

8.
通过大肠杆菌表达系统表达编码旋毛虫53kDa、49kDa ES基因重组蛋白P53、P49,这两种蛋白均可被感染旋毛虫(T.spiralis)小鼠阳性血清识别,应用重组蛋白P53和P49建立的两种间接ELISA检测感染鼠血清,探讨相关抗体消长规律及不同攻虫量血清抗体阳性时间差异.结果表明,血清相应抗体水平随感染时间的延长而增高,分别于27 d和31 d达到峰值,之后进入稳定期;不同数量T.spiralis攻虫后,各时期抗体阳性检出率P49-ELISA均优于P53-ELISA.综合敏感性和特异性,P49基因重组蛋白的敏感性和特异性更为适合用作检测旋毛虫病的理想抗原,并可应用于早期诊断,适合用于ELISA检测试剂盒的建立.  相似文献   

9.
通过在小鼠体内感染猪、犬旋毛虫肌幼虫50条,再用同源免疫和交叉免疫方法攻击感染旋毛虫肌幼虫200条,来检测其肌幼虫的减虫率。试验得到猪旋毛同源免疫的减虫率为87.47%,对犬旋毛虫的交叉免疫减虫率是86.47%,犬旋毛虫同源免疫的减虫率为83.70%,其对猪旋毛虫的交叉免疫是83.46%。试验表明:旋毛虫肌幼虫自然免疫具有较好的免疫保护作用,交叉免疫的保护率低于同源免疫。  相似文献   

10.
旋毛虫各隔离种雌虫体外产新生幼虫能力的研究   总被引:5,自引:0,他引:5  
本试验对旋毛虫各隔离种雌虫体外产新生幼虫能力进行了研究.结果显示,猪旋毛虫和旋毛形线虫(Trichinellaspiralis)雌虫体外培养24小时,平均产新生幼虫数分别为66.00±7.34和76.20±7.57,而犬旋毛虫和本地毛形线虫(Trichinellanatiua)分别是28.80±4.30和22.00±3.22,前者在雌虫体外产幼虫能力上明显高于后者.研究结果表明,黑龙江猪旋毛虫相当于旋毛形线虫,犬旋毛虫相当于本地毛形线虫.  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.
14.
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   

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.
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.
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.
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.
<正>1.INTRODUCTION Ensuring transportation network security is one Of the most daunting challenges confronting homeland security agencies today.Significant research has been dedicated.To model and analyze the vulnerability of transportation systems,while notably fewer studies propose specific strategies for deploying defensive technologies to safeguard these systems.  相似文献   

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