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为了制备兔抗鸽IgG抗体-HRP,建立斑点酶联免疫吸附试验定性检测鸽毛滴虫抗原的检测方法,将纯化的鸽IgG加入佐剂免疫兔子,获取纯化的兔抗鸽IgG抗体;采用简易过碘酸钠法标记兔抗鸽IgG,获取兔抗鸽IgG抗体-HRP;同时将鸽毛滴虫用超声波粉碎加入佐剂后多次免疫健康鸽获取鸽抗鸽毛滴虫高免血清(一抗);然后在2×106个/mL的鸽毛滴虫抗原量下,用不同浓度的一抗抗体、兔抗鸽IgG抗体-HRP进行试验;并对70个临床样品进行检测。结果显示,在2×106个/mL的鸽毛滴虫抗原量下,一抗与兔抗鸽IgG-HRP最佳工作浓度分别为1∶100和1∶500;70个样品中,48个镜检为阳性的样品用Dot-ELISA检测有44个呈阳性,阳性符合率为91.7%;22个镜检为阴性的样品用Dot-ELISA检测有12个呈阴性,阴性符合率为54.5%;Dot-ELISA检测与镜检结果的总符合率为80%。结果表明,本试验成功地建立了鸽毛滴虫感染的Dot-ELISA检测方法。 相似文献
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The poultry red mite Dermanyssus gallinae is a major pest and widespread ectoparasite of laying hens and other domestic and wild birds. Under optimal conditions, D. gallinae can complete its lifecycle in less than 10 days, leading to rapid proliferation of populations in poultry systems. This paper focuses on developing a theoretical model framework to describe the population dynamics of D. gallinae. This model is then used to test the efficacy and residual effect of different control options for managing D. gallinae. As well as allowing comparison between treatment options, the model also allows comparison of treatment efficacies to different D. gallinae life stages. Three different means for controlling D. gallinae populations were subjected to the model using computer simulations: mechanical cleaning (killing once at a given time all accessible population stages), sanitary clearance (starving the mite population for a given duration, e.g. between flocks) and acaricide treatment (killing a proportion of nymphs and adults during the persistence of the treatment). Simulations showed that mechanical cleaning and sanitary clearance alone could not eradicate the model D. gallinae population, although these methods did delay population establishment. In contrast, the complete eradication of the model D. gallinae population was achieved by several successive acaricide treatments in close succession, even when a relatively low treatment level was used. 相似文献
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旨在研究鸽毛滴虫是否分泌外泌体及虫源外泌体的蛋白组成和功能,为探索外泌体在鸽毛滴虫寄生过程中的作用奠定基础。采集鸽毛滴虫感染个体口腔及咽部病灶处样本,进行分离培养及纯化,使用显微镜鉴定虫体形态,提取DNA并比对其序列,确定培养物是否为鸽毛滴虫虫株;通过对培养液上清进行超速离心,透射电子显微镜、Western blot技术和纳米颗粒追踪分析（nanoparticle tracking analysis,NTA）鉴定培养液上清中是否含有虫源外泌体;最后通过Label-free技术分析虫源外泌体的蛋白质谱表达。结果显示,虫体具有明显的毛滴虫形态及特征,与鸽毛滴虫ITS1/5.8S/ITS2基因比对率达98%。从虫体培养液中提取的囊泡经透射电子显微镜观察具有明显的茶托样结构;NTA结果显示,粒径集中于125.1 nm,峰值粒径为132.3 nm,占比99.3%;CD63和TSG101等外泌体标记蛋白的条带明显,表明提取物为外泌体。质谱结果显示,烯醇酶、3-磷酸甘油醛-脱氢酶、磷酸甘油酸变位酶和延伸因子为表达量较高的蛋白质。GO功能注释显示外泌体蛋白富集到细胞内和胞浆等细胞组分,发挥GTP连接和GTP酶活性功能,参与小GTPase介导的信号转导与糖酵解等过程。KEGG通路富集表明,虫源外泌体蛋白主要富集于代谢途径、糖酵解/糖异生、抗生素的生物合成和次级代谢产物的生物合成等通路。本研究发现鸽毛滴虫可以分泌外泌体,并对虫源外泌体进行蛋白质谱分析,提示虫源外泌体中的蛋白质在寄生虫的能量代谢、信号转导及生物合成等过程中发挥重要作用。 相似文献
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曹小明 《湖南农业大学学报(自然科学版)》1990,(4)
研究鸡异刺线虫在罗斯鸡体内的发育史表明,感染性虫卵被宿主吞食后,卵内L_2于第4h开始释出,8～12 h内到达盲肠中发育,第4,6和12 d分别形成L_3,L_4和L_5,经31～56 d成熟排卵,此外,幼虫在肠粘膜内无组织移行过程。 相似文献
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David Harrington Hatem Mohi El Din Jonathan Guy Karen Robinson Olivier Sparagano 《Veterinary parasitology》2009,160(3-4):285-294
Dermanyssus gallinae is the most significant ectoparasite of European poultry egg laying production systems due to high costs of control and associated production losses as well as adverse effects on bird welfare. In this study, soluble proteins were extracted from unfed D. gallinae (DGE) using a urea-based detergent and ultra-filtration, passed through a 0.22 μm filter and blended aseptically with adjuvant. One group of laying hens was immunized with DGE and adjuvant (Montanide ISA 50 V) whilst another group (Control) received physiological saline and adjuvant. All birds were immunized on two occasions, 21 days apart. Antibody response to immunization was determined by ELISA and western blotting using immunoglobulins (Igs) extracted from egg yolk. DGE immunization of hens resulted in a significant (P < 0.05) IgY response compared to controls, although there was no significant difference in IgM response between treatments. A number of proteins were identified by western blotting using IgY antibodies from DGE immunized birds, most prominently at 40 and 230 kDa. Analysis of proteins from approximately corresponding bands on SDS-PAGE confirmed the identity of tropomyosin, whilst other proteins showed high sequence homology with myosin and actin from other arachnid and insect species. Immunization of hens with DGE resulted in a 50.6% increase in mite mortality (P < 0.001) 17 h after feeding when tested by an in vitro mite feeding model. Data in this study demonstrate that somatic antigens from D. gallinae can be used to stimulate a protective immune response in laying hens. Further work is needed to identify other proteins of interest that could confer higher protection against D. gallinae, as well as optimization of the vaccination and in vitro testing protocol. 相似文献
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The effect of neem oil (azadirachtin), originating from the tree Azadirachta indica, was investigated as a potential compound to control the poultry red mite, Dermanyssus gallinae. In vitro tests were performed to determine the most appropriate formulation of neem extracts and concentration of the substance to be used. Cardboard traps containing 20% neem oil were placed at the mites’ aggregation sites, out of reach of the hens, in a floor system for layers containing approximately 2400 birds. Treated traps were replaced every week for 4 weeks. Throughout the study, the parasite population was monitored by collections of mites with untreated plastic traps. A 92% reduction of D. gallinae was recorded. 相似文献