Studies on mutlispecific resistance of gastrointestinal nematodes to benzimidazoles on dairy-goat farms

Multispecific resistance to benzimidazoles was studied in three selected farms. These farms had bred dairy goats for more than 15 years. The helminths were introduced with the goats at the establishment of the farms which afterwards remained isolated. Nematode resistance could then be related to their own management practices. Faecal egg count tests and egg hatch assays were performed to assess intensity of resistance. The generic (infective larvae in faecal cultures) and specific richness (adult worms) were assessed. The resistant species were Trichostrongylus colubriformis, Teladorsagia circumcincta, Haemonchus contortus and Oesophagostomum venulosum. Faecal egg count reduction tests and egg-hatch assays did not match exactly. Faecal larval counts after treatments gave a distorted picture of multispecific resistance: Haemonchus and Oesophagostomum were very largely over represented. The number of species found in the three farms was relatively low compared with other reports in goat farms of the area. This reduction of diversity might also be due in part to characteristics of breeding management and history (use of permanent pasture and introduction of goats at the establishment of farm).     

应用计算机和气象资料预测粗纹食道口线虫（Oesophagostomum asperum）数量季节消长的研究

根据土源性寄生线虫体外发育期间受温度、湿度、季节和宿主动物影响的原理，采用电子计算机对上述四因子和食道口线虫从卵发育到侵袭性幼虫速度、数量进行回归分析，制定出模拟模型，设计出了在牧地上该种线虫从卵发育到侵袭性幼虫数量预测预报公式，用该公式和气象资料计算出的侵袭性幼虫季节消长曲线，与牧地上宿主羊体内成虫数量消长曲线相一致，初步显示可以采用气象资料和计算机预测牧地上粗纹食道口线虫侵袭性幼虫量的季节动态，从而为防治措施的制定提供依据。     

The in vitro motility response to various anthelmintics of third-stage larvae of Oesophagostomum spp. from pigs

The in vitro activities of thiabendazole, levamisole, pyrantel, morantel and ivermectin against Oesophagostomum spp., the nodular worm of pigs, were determined and compared. The study was carried out using isolates of O. dentatum and O. quadrispinulatum, which had been defined in vivo. Infective larvae were exposed to the anthelmintics for 24 h and then placed in a micromotility meter. All the treatments significantly reduced the motility of the ensheathed L₃ larvae, but the micromotility meter was not able to differentiate between anthelmintic resistant and anthelmintic susceptible isolates.     

有齿食道口线虫ITS及5.8S DNA片段的PCR扩增、克隆及序列分析

 运用PCR方法以保守引物NC5及NC2扩增了从广东阳江地区猪体分离的食道口线虫rDNA 的内转录间隔区（ITS）及5.8S序列。将PCR扩增出的片段纯化后克隆至pGEM-T Easy载体，用PCR技术及酶切鉴定阳性菌落，对阳性菌落质粒DNA进行测序。结果表明，扩增的片段大小为828 bp，包含部分的18S、28S及全部的ITS-1（362 bp）、5.8S(153 bp)及ITS-2（217 bp）序列。序列比较表明，该食道口线虫为有齿食道口线虫。本研究在国际上首次报道了中国猪有齿食道口线虫的ITS及5.8S序列，为食道口线虫的分子生物学的进一步研究奠定了基础。     

绵羊寄生线虫在体内及牧场上各发育阶段的季节动态研究

对亚热带草地模式区──湖北长阳火烧坪草畜示范场绵羊寄生线虫的优势种捻转血矛线虫、环纹奥斯特线虫和粗纹食道口线虫进行了成虫寄生量、羊粪中虫卵排出量和寄生虫体外发育育成率的逐月观察，结合气象资料──气温、相对湿度、蒸发量和雨量的观察记录，得出了这些优势种各发育阶段在绵羊体内及牧地上的季节动态以及与气温、湿度变化的关系。３种线虫在牧地上的体外育成率和在羊体内的寄生量都有２个峰，育成率于４月出现第１个峰，随之在５月成虫出现第１个峰，育成率于６或７月分别出现第２个更高的峰，成虫也于７～９月出现第２个更高的峰。说明成虫峰随体外发育育成峰而来，粪中虫卵的峰值则往往跟随成虫峰而出现。体外育成率的高低又明显受气温及温度影响，平均湿度低于１２℃时无峰出现，低于５℃时育成率为零，湿度在７９％以下时育成率很低。本文还讨论了防制建议。     

分子鉴定2种猪食道口线虫熔解曲线分析试验的建立

基于猪有齿食道口线虫和四棘食道口线虫IGS rDNA序列的遗传变异情况,于种内保守、种间差异明显的序列片段设计1对引物,建立了区分2种猪食道口线虫的熔解曲线分子鉴定方法。研究结果显示,该方法可通过熔解曲线差异而准确鉴定猪食道口线虫2个种,与其他线虫和吸虫无交叉反应,特异性良好;能检测猪食道口线虫单虫卵样品和FTA卡片的DNA模板。该研究为猪食道口线虫种类准确鉴定提供了一种新的准确、敏感、特异且快速的分子手段。     

Development and survival of infective larvae of gastrointestinal nematodes of cattle on pasture in central Kenya

On a series of pasture plots, 2 kg pats of bovine faeces containing known numbers of strongylid (Haemonchus, Cooperia, Oesophagostomum and Trichostrongylus) eggs were deposited at intervals of 4 weeks from July 1995 to June 1996. The plots were sampled every 2 weeks after contamination and infective larvae were identified and counted. Larvae of all the genera developed throughout the year, but the pats exposed during the rainy season yielded more abundant larvae on the herbage. Irrespective of the season of deposition of the pats, larvae were found in larger numbers from 2 to 6 weeks after deposition and generally declined to below detectable levels within 12 to 16 weeks of contamination. The comparatively short survival times noted in this experiment may present opportunities for manipulation of the population dynamics of the gastrointestinal nematodes in the tropical environment of Kenya.     

羊体的三种线虫在不同温湿度条件下从卵到侵袭性幼虫发育能力的观察

对寄生于羊的捻转血矛线虫、粗纹食道口线虫、环纹奥斯特线虫在不同恒温和恒湿条件下从卵发育到侵袭性幼虫的育成率和所需时间作了观察，温度有４０、３７．５、３５、３０、２５、２０、１５、１０、７．５和５℃等１０个段，相对湿度有１００、９０、８５、８０、７５％等５个相。结果捻转血矛线虫仅在１０～３５℃温度和８０％以上相对湿度条件下能完成发育，１００％相对湿度和２０℃条件下育成率最高；粗纹食道口线虫和环纹奥斯特线虫在７．５～３７．５℃和相对湿度８０％以上能完成发育，也是在１００％湿度和２０℃条件下育成率最高。试验结果可供研究这些寄生虫的地理分布和制定其防治措施时作依据。     

中国南方山地羊消化道线虫体外发育的研究

本文检测中国南方亚热带山地环境因素（温度，相对湿度和降雨量）变化对羊捻转血矛线虫、环纹奥斯特线虫和粗纹食道口线虫体外发育的影响规律：捻转血矛线虫由卵至Ⅰ、Ⅱ和Ⅲ期幼虫的最高育成率均在６月；育成期分别是３、５和７ｄ。环纹奥斯特线虫卵至Ⅰ、Ⅱ和Ⅲ期幼虫育成率在５～７月均较高，其中Ⅰ和Ⅱ期幼虫最高育成率在６月，而Ⅲ期幼虫最高育成率在５月；育成期分别是３、５和１１ｄ。粗纹食道口线虫卵至Ⅰ、Ⅱ和Ⅲ期幼虫的最高育成率均在７月；育成期分别是６、７和９ｄ。１１月至次年３月间三线虫均不能发育，而且粗纹食道口线虫于１０月停止发育。文中还讨论了三线虫的防治措施。     

Evidence for multiple anthelmintic resistance in sheep and goats reared under the same management in coastal Kenya

Four experiments, two with sheep and two with goats, were carried out to determine the efficacy of ivermectin, fenbendazole, levamisole, closantel and some of their combinations by faecal egg count reduction tests. In the first experiment, injectable ivermectin, oral ivermectin, fenbendazole and levamisole were tested in 6-month-old lambs, and their reduction percentages were 77%, 13%, 42% and 92%, respectively. In the second experiment, with yearling sheep, the reduction percentages were 35% for injectable ivermectin, 32% for fenbendazole, 99% for levamisole, 48% for closantel, 92% for injectable ivermectin combined with fenbendazole, 99% for injectable ivermectin combined with levamisole, and 100% for fenbendazole combined with levamisole. In the study with 18-month-old goats given the same dose rates as those recommended for sheep, the reduction percentages were 73% for injectable ivermectin, 25% for fenbendazole, and 78% for levamisole. Another group of 14-month-old goats was treated with dose rates 1.5 times those recommended for sheep and the reduction percentages were 93% for levamisole, 92% for injectable ivermectin, and 97% for a combination of levamisole and ivermectin. In all experiments with sheep and goats the gastrointestinal nematode parasites identified by larval cultures were Haemonchus contortus, Trichostrongylus spp. and Oesophagostomum spp. The gastrointestinal nematodes of both sheep and goats on this farm are resistant to ivermectin and fenbendazole, whereas levamisole is still effective in sheep, but not in goats. The results are discussed in relation to the farm as a source of breeding stock to smallholder farmers and its potential to spread anthelmintic resistance.