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1.
捻转血矛线虫感染性三期幼虫对干燥环境具有很强的抵抗力,环境适宜时可恢复到正常状态并继续感染宿主.为初步探讨捻转血矛线虫感染性三期幼虫抗干燥的分子机制,试验对该期幼虫施以不同干燥条件的处理,筛选出虫体存活率达到90％的干燥条件.在筛选到的条件下(相对湿度为50％,时间为60 h)处理虫体,采用mRNA差异显示技术分析干燥环境下培养的虫体和正常虫体的mRNA表达差异.经验证获得了48个差异表达的EST序列,其中25个(52.08％)序列与现有捻转血矛线虫基因组数据库有高度同源性；15个(31.25％)序列与已知线虫的cDNA序列或基因组序列有较高同源性；9个(18.75％)序列与其他线虫已知蛋白质氨基酸序列同源性较高.差异表达序列AF-U01C与多种线虫的蛋白酶体β亚基家族成员同源性达90％,差异表达序列AG-U01A与多种线虫的β酮脂酰CoA硫解酶(3-ketoacyl-CoA thiolase)同源性达65％.结果为进一步研究寄生性线虫抗干燥机制奠定基础. 相似文献
2.
为研究干燥作用对捻转血矛线虫L3期幼虫mRNA表达的影响,筛选并分析干燥相关表达基因,本试验利用mRNA差异显示技术得到了捻转血矛线虫干燥相关基因,通过相对荧光定量PCR验证所获基因的表达情况,并在秀丽隐杆线虫中通过RNA干扰抑制相关同源基因的表达检测干燥后复苏虫体生存率以研究基因抗干燥功能。结果显示,差异表达序列GH-U02A与已知捻转血矛线虫谷胱甘肽-S-转移酶(GST)氨基酸同源性为94%。对秀丽隐杆线虫Ce-gst-7基因进行RNA干扰后,虫体在干燥后的生存率降低,证明gst基因在线虫中有抗干燥功能。 相似文献
3.
选取捻转血矛线虫脂肪酶(Lipase)进行基因克隆、表达并对其生物学特性进行探究。PCR克隆捻转血矛线虫lipase基因,构建表达载体,获得融合蛋白质,采用Western blot检测该蛋白质的抗原特性;酯类酶解反应检测该融合蛋白质脂肪酶活性;实时定量PCR检测该基因在捻转血矛线虫不同发育阶段的转录情况;间接免疫荧光试验检验该抗原蛋白质在虫体组织的定位情况。结果表明,去除lipase基因信号肽序列后,基因片段长864bp,融合蛋白质大小50ku;该融合蛋白质在37℃左右、pH为7的条件下酶活力最高;雄性成虫lipase转录量相对虫体其他阶段最高;该抗原蛋白质主要表达于虫体表皮和肠腔。Lipase具有较好的抗原性,可能与捻转血矛线虫发育和宿主免疫调节有关,深入研究捻转血矛线虫Lipase的特性对寻找新的免疫保护性抗原和药物靶标具有意义。 相似文献
4.
捻转血矛线虫(Haemonchus contortus)氨基肽酶基因克隆、表达及重组蛋白活性分析 总被引：1，自引：0，他引：1
通过RT—PCR扩增出捻转血矛线虫（Haemonchus contortus）氨基肽酶基因，并对该基因进行克隆、鉴定及序列测定。捻转血矛线虫氨基肽酶ORF由2919个碱基（972个氨基酸）构成，和GenBank中的捻转血矛线虫氨基肤酶（H11抗原）同源性高达98％。与秀丽新杆线虫（Caenorhabditis elegans）肽酶M1家族同源性为61％，且具有氨基肽酶特征性基序HEXXH和GAMEN。将该基因亚克隆到原核表达栽体并进行了表达，通过测定重组蛋白分解氨基肽酶底物的能力以及对酶抑制荆敏感性试验，进一步证实了H11抗原具有一定的酶活性。 相似文献
5.
为进一步研究寄生性线虫的滞育机制,参照Adams(1986)的方法,通过瘤胃灌注,将捻转血矛线虫感染性三期幼虫攻入绵羊体内,对绵羊进行处理后,获得捻转血矛线虫滞育虫体。滞育虫体发育程度一致,虫体长(1 067±97)μm(n=18),肠道细胞中含有大量棒状结晶块;性腺开始发育,出现雌驱器原基;颈部出现颈乳突。通过温度检测分析,温度可能不是滞育现象的主要刺激因素。棒状结晶块可以被DAPI特异性着色,其成分可能含有核酸。本试验将为进一步研究捻转血矛线虫的滞育机制及其防治奠定基础。 相似文献
6.
《中兽医医药杂志》2016,(5)
为了对捻转血矛线虫进行分子鉴定,并且探寻其与毛圆科内不同属线虫存在差异的分子水平关联性,对采自牦牛体内的捻转血矛线虫线粒体COXⅠ基因部分序列进行扩增与进化分析,扩增出的序列登录到GenBank,登录号:KU314701。进化分析结果显示,捻转血矛线虫H-yak线粒体COXⅠ基因与毛圆科不同属线虫的线粒体COXⅠ基因同源性在85.5%~97.4%之间,其中与KJ724363.1的同源性最高,达到97.4%;其编码的蛋白氨基酸序列与血矛属线虫的线粒体细胞色素C氧化酶亚基Ⅰ蛋白的氨基酸序列完全相同,同源性为100%,且与长刺属线虫的同源性极高,同为100%,但与古柏属和奥斯特属的线虫蛋白同源性均为98.7%。该研究在基因和蛋白两个层面间接印证了毛圆科内不同属线虫在形态和生活史方面有较多相似和不同之处。 相似文献
7.
旨在解析3种不同形态阴门盖捻转血矛线虫（Haemonchus contortus）的遗传进化差异，本研究从养殖场山羊皱胃中分离到捻转血矛线虫虫体，采用普通光学显微镜观察71条雌虫虫体阴门盖的形态结构特征，并基于捻转血矛线虫ITS-1、ITS-2和nad4基因位点，采用PCR法对不同阴门盖的虫体进行扩增，通过比对多位点序列进行遗传进化分析。经显微镜观察发现，本研究分离到的捻转血矛线虫雌虫阴门盖存在舌瓣形、亚球形和舌-球混合形3种形态，所占比例分别为45.07%（32/71）、50.70%（36/71）和4.23%（3/71）。对43条雌虫全长和阴门至虫体末端长度进行测量和统计分析，结果显示，3种不同形态阴门盖的捻转血矛线虫全长统计学差异不显著（P>0.05），而3种不同形态阴门盖的捻转血矛线虫阴门至虫体末端的长度存在显著性差异（P<0.05）。基于捻转血矛线虫ITS-1、ITS-2和nad4基因进行的序列分析结果显示，不同形态阴门盖虫体的9个样品在3个基因位点上存在不同程度的碱基差异，多态性位点分别为10、13和43个，核苷酸多态性分别为0.85%、1.34%和1.97%。基于ITS-1和ITS-2构建的遗传进化树显示，研究中所用样品与GenBank上H.contortus的参考序列处于同一进化支，表明本研究所分离到的虫体均属捻转血矛线虫。捻转血矛线虫雌虫的阴门盖存在形态学差异，不同形态阴门盖虫体在ITS-1、ITS-2和nad4基因位点上均存在不同程度的碱基差异，此研究结果为捻转血矛线虫的种类鉴定和遗传进化提供了参考数据。 相似文献
8.
《中国兽医学报》2019,(12):2342-2349
捻转血矛线虫(Haemonchus contortus)是最重要的家畜寄生虫之一。已报道秀丽隐杆线虫(Caenorhabditis elegans)转甲状腺素蛋白(transthyretin,TTR)参与虫体细胞间的相互作用。目前捻转血矛线虫转甲状腺素蛋白(Hc-TTR-51)的定位及功能尚未见报道。研究首先克隆捻转血矛线虫Hc-ttr-51基因,然后分析其蛋白在捻转血矛线虫、HEK 293T细胞内的定位情况,最后RNA干扰Hc-ttr-51基因分析对秀丽隐杆线虫生长发育的影响,从而对其功能进行预测分析。结果表明,捻转血矛线虫Hc-ttr-51基因在虫体各发育阶段均有表达,与其他发育期虫体相比,L1、L3、雄成虫该基因的转录水平显著上升。在L3期虫体体内该蛋白广泛表达,L4期和成虫体内该蛋白特异性表达于性腺。在HEK 293T细胞中,该蛋白在细胞质中呈明显的点状分布。RNA干扰秀丽隐杆线虫Hc-ttr-51基因表达后,虫体产卵量极显著下降,体长体宽显著缩短,咽泵率显著下降。以上研究为捻转血矛线虫生长发育特性、疫苗研究等提供依据。 相似文献
9.
捻转血矛线虫ZJ株24000分泌排泄(ES)蛋白基因的克隆和序列分析 总被引：1，自引：0，他引：1
通过 RT- PCR方法 ,从捻转血矛线虫成虫总 RNA中扩增得到特异性片段 ,并把这一基因片段克隆到 p U C18克隆载体上 ,进行测序及同源性比较。序列分析可知 ,其核苷酸序列与国外已发表的 2 4 0 0 0分泌排泄抗原基因的同源性为 99.2 %。表明本试验成功提取了捻转血矛线虫成虫总 RNA,并用 RT- PCR方法克隆了 2 4 0 0 0分泌排泄抗原基因 ,为该基因的表达及其免疫学作用研究奠定了基础 相似文献
10.
为鉴定绵阳地区山羊粪便中疑似血矛属线虫虫卵的虫种,本研究提取绵阳地区三个县山羊粪便中疑似血矛属线虫的虫卵DNA 60份,采用PCR技术扩增其核糖体第二内转录间隔区(rDNA ITS-2)基因序列,并与GenBank上公布的捻转血矛线虫同源序列进行比较分析。结果显示:60份疑似血矛属线虫的虫卵DNA样品均得到191bp的ITS-2片段;60条ITS-2序列共形成15条不同的序列,含有17个单变异位点,与GenBank中多株捻转血矛线虫的同源性达95.8%~100%;ML系统发育树显示,本研究得到的15种类型ITS-2序列与已报道的捻转血矛线虫(Haemonchus contortus)聚类形成一个分支。综上可得,60份来自山羊的血矛属线虫均为捻转血矛线虫。 相似文献
11.
The effects of both natural and artificial conditioning of pre-infective and infective stages of Haemonchus contortus were examined in two studies with 108 crossbred lambs, using an Ohio isolate of H. contortus known to exhibit up to 100% winter hypobiosis. The intensity of hypobiosis varied from 0 to 36% in 17 lamb groups given larvae subjected to various temperature and photoperiod conditions. The results in lambs given larvae conditioned for 4-8 weeks were not significantly different from those in lambs given freshly cultured larvae. It is suggested that winter hypobiosis of H. contortus in the northern U.S.A. is an obligatory survival mechanism that occurs without the need for external stimuli to trigger the onset of hypobiosis. 相似文献
12.
J Vercruysse 《Veterinary parasitology》1985,17(2):159-163
Throughout a period of 13 months, the abomasa of 1024 sheep, living in a sahelian region, were examined for adult Haemonchus contortus and their inhibited L4 larvae. Of these 796 (78%) were found harbouring H. contortus, but with low worm burdens (geometric mean 37). Significant numbers of inhibited larvae appeared early in the dry season (November) and they represented 49% of the total in December. A similar proportion of inhibited larvae were present until April and after that their number declined abruptly. It was concluded that Haemonchus sp. survives the dry season as hypobiotic larvae as well as adults. 相似文献
13.
R K Reinecke H M Snyman H Seaman 《The Onderstepoort journal of veterinary research》1979,46(4):199-205
Fifty-thousand infective larvae of Trichostrongylus axei dosed to 7-9-month-old Merinos protected them against a subsequent challenge with 50 000 infective larvae of Haemonchus contortus. Protection varied with the period between an initial dosage of T. axei and challenge with H. contortus as follows: After 90 days it was more than 80% effective in more than 80% of sheep; at 119 and 180 days there was still a significant reduction in the numbers of H. contortus recovered (P < 0,05 and P < 0,001, respectively). Prior infestation with either Ostertagia circumcincta or with Haemonchus placei was ineffective against challenge with H. contortus. 相似文献
14.
Two groups of goats were experimentally infected with two different strains of Haemonchus contortus and compared with the controls. Group A animals were infected with a goat-derived strain (GDS) while Group B animals were infected with a sheep-derived strain (SDS). Changes in the liveweights and some blood constitutents between the two infected groups were compared. The pathogenic effects of the GDS larvae were generally more serious when compared with those of the SDS larvae. 相似文献
15.
Partial protection of lambs against Haemonchus contortus by vaccination with a fractionated preparation of the parasite 总被引：1，自引：0，他引：1
Florida Native lambs, less than 6 months of age, were successfully vaccinated against Haemonchus contortus with a high mol. wt fraction (greater than 30,000 daltons) derived from a somatic extract of H. contortus larvae (SEL) and excretions and secretions (ES) of larvae isolated during in vitro development from the infective 3rd to 4th stage. A 59% reduction in adult worm numbers was obtained in vaccinates compared to naive lambs following challenge. The protection in vaccinated lambs was similar to that seen in lambs exposed to a primary infection of H. contortus larvae which had been cleared with anthelmintic prior to the challenge infection. The unfractionated SEL/ES preparation and a low mol. wt fraction gave no significant protection against challenge infection. 相似文献
16.
R J Boisvenue M I Stiff L V Tonkinson G N Cox R Hageman 《American journal of veterinary research》1992,53(7):1263-1265
Sixteen nonsibling sheep, approximately 12 months old, that were raised in a helminth-free environment, were used for 2 protection studies 6 months apart. Sheep were vaccinated weekly for 5 weeks by IM injection of fibrinogen-degrading proteins derived from the intestinal tract of adult Haemonchus contortus. Ten days after the last vaccination, sheep were given 2,500 infective H contortus larvae by intraruminal injection. Vaccinated sheep produced specific antibodies, and were protected from the worm challenge. Significant differences in mean fecal worm egg counts for 56 days after worm challenge, in mean numbers of H contortus worms, and female fecundity ratios at necropsy were detected in vaccinated sheep, compared with those in control sheep. These data suggest that the fibrinogen-degrading proteins have a protective role in vaccination of sheep against H contortus. 相似文献
17.
J E Miller F G Hembry M T Kearney J C Williams L C Stagg D Sims 《American journal of veterinary research》1987,48(9):1403-1406
Efficacy of levamisole was evaluated in a suspected levamisole-resistant population of Haemonchus contortus in the Louisiana State University sheep flock. The efficacy of netobimin also was evaluated against this population of Haemonchus. In trial 1, 5 lambs naturally infected with H contortus were given 8 mg of levamisole/kg of body weight as a drench, and 5 lambs were not treated (controls). Nematode recovery after slaughter indicated 0% efficacy against H contortus. In trial 2, 30 nematode-free lambs were each given 8,300 F1 generation infective larvae of H contortus, which were derived from parent H contortus that survived 2 levamisole treatments in lambs being maintained in a nematode-free environment. Ten lambs were treated with 8 mg of levamisole/kg as a drench, 10 were treated with 20 mg of netobimin/kg as a drench, and 10 were not treated (control). Nematode recovery after slaughter revealed 62.3% and 99.8% efficacy for levamisole and netobimin, respectively, against H contortus. 相似文献
18.
Decreased ivermectin and moxidectin sensitivity in Haemonchus contortus selected with moxidectin over 14 generations. 总被引：1，自引：0，他引：1
Ivermectin resistance in the nematode Haemonchus contortus has been reported in many parts of the world and many ivermectin resistant isolates have been found to have reduced sensitivity to moxidectin. However, it is unclear whether parasites that are selected with moxidectin would demonstrate reduced sensitivity to ivermectin. In this study, the effects of moxidectin and ivermectin on an unselected strain and a strain of H. contortus derived from the unselected strain but selected over 14 generations with moxidectin, were compared in jirds. The recovery of adult worms and fourth stage (L4) larvae following treatment were compared between strains and anthelmintics. Moxidectin-selected H. contortus showed reduced sensitivity to ivermectin as well as to moxidectin. Doses of 0.1 mg/kg of moxidectin and 0.4 mg/kg of ivermectin were necessary to obtain an efficacy of 95% or above against the moxidectin-selected strain of H. contortus compared with 0.025 mg/kg for moxidectin and 0.1 mg/kg for ivermectin required for a similar efficacy in the unselected strain. 相似文献
19.
Effect of copper oxide wire particle treatment on establishment of major gastrointestinal nematodes in lambs 总被引：1，自引：0，他引：1
Eighty-four 10-week-old lambs were used to investigate the effect of copper oxide wire particle treatment on the establishment of major gastrointestinal nematodes. They were maintained on pasture previously treated to minimise larval contamination. Five grams of uniform sized copper oxide wire particles were given orally five days before infection with either 20,000 Trichostrongylus colubriformis larvae, 20,000 Ostertagia circumcincta larvae or 3000 Haemonchus contortus larvae given as three doses at three-day intervals. The animals were slaughtered 21 or 22 days after the last infective dose. Parasite burdens in the lambs treated with copper oxide wire particles were reduced by 96 per cent in the case of H contortus and by 56 per cent in the case of O circumcincta compared to burdens in controls. There was no significant effect of copper particles on the establishment of T colubriformis. 相似文献