猪食道口线虫ITS-1和lTS-2 rDNA的PCR-SSCP分析

以采自我国不同地区猪体的食道口线虫虫株为研究对象,PCR扩增出ITS-1和ITS-2序列片段,然后采用单链构象多态性(SSCP)方法分析PCR产物,对不同地区食道口线虫进行分子鉴定。所有样品经SSCP分析显示两种带型,第一种为有齿食道口线虫带型,另一种为未定种食道口线虫带型。代表性样品的测序结果表明,未定种食道口线虫带型的样品为四棘食道口线虫。本研究在国际上首次报道了中国猪四棘食道口线虫的ITS序列,并建立了区分有齿食道口线虫和四棘食道口线虫的PCR-SSCP方法,从而为食道口线虫的分子生物学的进一步研究奠定了基础。     

猪食道口线虫ITS—1和ITS-2rDNA的PCR—SSCP分析

以采自我国不同地区猪体的食道口线虫虫株为研究对象，PCR扩增出ITS-1和ITS-2序列片段，然后采用单链构象多态性（SSCP）方法分析PCR产物，对不同地区食道口线虫进行分子鉴定。所有样品经SSCP分析显示两种带型，第一种为有齿食道口线虫带型，另一种为未定种食道口线虫带型。代表性样品的测序结果表明，未定种食道口线虫带型的样品为四棘食道口线虫。本研究在国际上首次报道了中国猪四棘食道口线虫的ITS序列，并建立了区分有齿食道口线虫和四棘食道口线虫的PCR-SSCP方法，从而为食道口线虫的分子生物学的进一步研究奠定了基础。     

连续驱虫法防治猪食道口线虫病试验报告

尽管驱除猪食道口线虫的药物较多，但食道口线虫仍然给养猪业造成巨大的经济损失。根据苏联学者报道；单独感染食道口线虫的70天仔猪增重比对照组猪少17．55％。另据报道，食道口线虫在美国、加拿大和世界其它地区，都是经济上重要的寄生虫，感染猪每头增重170磅所耗饲料成本比来感染猪多1．70美元。     

蛇四棘食道口线虫线粒体cox1 基因的克隆及序列分析

本研究旨在分析长沙市四棘食道口线虫分离株线粒体细胞色素c氧化酶第Ⅰ亚基(cox1)基因部分序列(pcox1)的遗传变异情况。应用聚合酶链反应(PCR)扩增四棘食道口线虫虫株的pcox1,应用Clustal X 1.83程序对序列进行比对,同时利用DNAStar 5.0中的MegAlign程序进行同源性分析,并与GenBank^TM中已知四棘食道口线虫相应基因序列进行比较分析。所得pcox1序列长度一致,均为393 bp,与GenBank公布的线虫相关序列进行比较分析结果表明,各个分离株与已知四棘食道口线虫相应基因的相似性分别在98%以上,与其它科线虫的相似性均小于91%。四棘食道口线虫pcox1序列种内相对保守,种间差异明显。本研究为进一步研究四棘食道口线虫的群体遗传学奠定了基础。     

羊消化道4种易混淆线虫虫卵的形态学观察与比较

为了方便临床上鉴定羊消化道常见且易混淆线虫虫卵并丰富相关资料,试验从羊消化道中采集疑似捻转血矛线虫、夏伯特线虫、食道口线虫和奥斯特线虫共4种线虫,采用形态学和分子生物学方法鉴定虫种,将4种线虫于体外短暂培养后得到纯种虫卵,通过观察虫卵不同发育阶段的胚胎形态变化获取每种线虫虫卵形态特征,并对虫卵的长径、宽径和长宽比进行测定。结果表明：经鉴定4种线虫分别为捻转血矛线虫、绵羊夏伯特线虫、粗纹食道口线虫和普通奥斯特线虫。4种线虫虫卵初期胚胎多呈收缩状态且形似桑葚,在外观形态上具有相似性,均为双层卵壳,多数对称,少数不对称。捻转血矛线虫虫卵呈短椭圆形,颜色较浅;绵羊夏伯特线虫虫卵呈椭圆形,颜色较捻转血矛线虫虫卵深;粗纹食道口线虫虫卵呈长椭圆形,颜色较捻转血矛线虫虫卵深,与绵羊夏伯特线虫虫卵相近;普通奥斯特线虫虫卵亦呈长椭圆形,颜色与捻转血矛线虫相近,较绵羊夏伯特线虫和粗纹食道口线虫虫卵浅。虫卵在生理盐水中培养1～6 d后,捻转血矛线虫、绵羊夏伯特线虫和普通奥斯特线虫依次进入蝌蚪期和幼虫期,但仅凭肉眼依然难以区分虫卵种类;绵羊食道口线虫未观察到含蝌蚪或含幼虫的虫卵,仅在桑葚期观察到胚胎收缩成1团及...     

应用计算机和气象资料预测粗纹食道口线虫（Oesophagostomunasper…

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，设计了在牧地上该种线虫从卵发育到侵袭性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和曲线相一致，初步显示可以采用气象资料和计算机预牧地上粗纹食道口线虫侵侵袭性幼虫量的季节动态     

多重PCR鉴定三种颚口线虫方法的建立

为了建立快速、灵敏、可靠的鉴别颚口线虫虫种的方法,根据GenBank中发表的棘颚口线虫、日本颚口线虫和杜氏颚口线虫ITS-2序列,设计了3对特异引物,建立了这3种颚口线虫的单一PCR和多重PCR检测方法,并分别对单一PCR和多重PCR方法的特异性和敏感性进行了鉴定。结果显示单一PCR和多重PCR均能特异扩增出棘颚口线虫、日本颚口线虫和杜氏颚口线虫,其片段大小分别为282、358、183 bp,单一PCR对棘颚口线虫、日本颚口线虫和杜氏颚口线虫虫体DNA最小检出量分别为0.2、0.01、0.01 ng/μL。对宫脂线虫、异尖线虫、棘口吸虫以及迭宫绦虫均不能进行扩增。用建立的多重PCR方法对菲律宾、印度尼西亚、黑龙江颚口线虫等12条颚口线虫DNA模板进行扩增,经鉴定菲律宾与印度尼西亚的颚口线虫为棘颚口线虫,黑龙江的颚口线虫为日本颚口线虫。鉴定结果与原虫体样本的形态鉴定和测序分析结果一致。研究显示建立的多重PCR方法具有很强的特异性和较高的敏感性,可用于棘颚口线虫、日本颚口线虫和杜氏颚口线虫虫种的鉴定和流行病学调查。     

阿维菌素在防治猪寄生虫病中的应用

1 驱虫范围 阿维菌素具有同时驱除猪内、外寄生虫的优点,它对内寄生虫的胃肠道线虫,如猪蛔虫、猪胃圆线虫、猪食道口线虫（结节虫）、猪毛首线虫（鞭虫）等的成虫和大部分的第四期幼虫以及肺线虫的猪后圆线虫（肺丝虫）、猪冠尾线虫（猪肾虫）的成虫都具有驱杀作用．但对它们的卵没有杀灭作用，阿维菌素对吸虫和绦虫、结肠小袋纤毛虫、猪球虫等没有作用。     

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。     

利用正交设计优化羊食道口线虫SRAP-PCR反应体系

本研究利用正交设计L16(4)对羊食道口线虫SRAP-PCR反应体系的4因素:Taq酶、Mg2+、dNTPs、引物进行摸索。结果表明:各因素的不同水平对SRAP-PCR反应结果都有显著的影响,正交设计组合4中扩增的条带最清晰;羊食道口线虫SRAP-PCR最佳反应条件:最佳引物给合为Me1/Em7,最佳反应体系为25μL:2.5μL 10×PCR buffer、20 ng模板DNA、Mg2+(25 mmol/L)3μL、dNTPs(2.5 mmol/L/μL)3μL、引物(10 pmol/μL)4μL、rTaq DNA聚合酶(5U/μL)0.2μL、灭菌ddH20补足。本研究结果为进一步采用SRAP技术来研究食道口线虫遗传进化奠定了坚实基础。     

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.     

A dose-response investigation on the level of resistance to pyrantel citrate in nodular worms of pigs

This study was undertaken to determine the level of resistance against pyrantel citrate in strains of Oesophagostomum quadrispinulatum and Oesophagostomum dentatum which have previously been found resistant to this anthelmintic. Groups of pigs were artificially infected with batches of infective larvae which were previously found either susceptible or resistant to pyrantel citrate. After treatment with 1, 2 and 4 times the recommended dose (14 mg kg-1) of pyrantel citrate, the resistant O. quadrispinulatum population was reduced by 51.0, 76.2 and 86.1%, and O. dentatum by 41.2, 47.9 and 78.5%. The results indicated that O. dentatum was slightly more resistant (P less than 0.05) than O. quadrispinulatum to pyrantel citrate. Treatment of the susceptible worms with 1 and 2 times the recommended dose caused a reduction in worm numbers by 61.0 and 99.4%, respectively.     

Resistance to levamisole and cross-resistance between pyrantel and levamisole in Oesophagostomum quadrispinulatum and Oesophagostomum dentatum of pigs

Two strains of Oesophagostomum spp., consisting of both O. quadrispinulatum and O. dentatum, were subjected to a controlled in vivo assay for resistance to levamisole and pyrantel by comparison with susceptible isolates. One strain (LEM) was recently isolated from a commercial herd, where sows showed high numbers of strongyle eggs in faeces within 2 weeks of farrowing and following treatment with levamisole at the manufacturer's recommended dose rate 1 week before farrowing. Levamisole had been used as the sole anthelmintic for treatment for at least 7 years on this farm. Treatment with pyrantel in this herd also indicated cross-resistance to this drug. A mixed population of O. quadrispinulatum and O. dentatum of this strain was subjected to controlled in vivo assays. Faecal egg count reduction (FECR) was found to be -573.3% (P greater than 0.05) and worm count reductions (WCR) of O. quadrispinulatum and O. dentatum were estimated as 44.5% (P greater than 0.05) and 96.4% (P less than 0.001), respectively. Treatment with pyrantel showed that cross-resistance existed to this drug, with FECR of 10.4% (P greater than 0.05) and WCR of 64.5% (P greater than 0.05) and 90.7% (P less than 0.05) for O. quadrispinulatum and O. dentatum, respectively. Another strain (VJ) was isolated from another commercial pig herd, which was dosed with pyrantel citrate four times a year for at least 8 years. This strain showed resistance to pyrantel, with FECR of 43.8% (P greater than 0.05) and WCR of 65.9% (P greater than 0.05) and 49.4% (P greater than 0.05) for O. quadrispinulatum and O. dentatum, respectively. However, both species were susceptible to levamisole. Our results suggested that selection with levamisole gave rise to levamisole resistance and automatically conferred resistance to pyrantel, whereas selection with pyrantel only resulted in resistance to this drug alone. These findings are discussed in relation to the location of the two species of Oesophagostomum in the large intestine of pigs and the mode of action of this class of anthelmintics.     

Transmission of Oesophagostomum Species in Swine on Pasture in the Maritime Provinces

Field experiments carried out in the Maritime area of Canada failed to demonstrate overwinter survival of porcine nodular worms, Oesophagostomum quadrispinulatum and O. dentatum. Pastures were contaminated by carrier animals with subsequent transmission of infection to susceptible pigs. One approach to control of oesophagostomiasis in grazing swine is the treatment of all infected pigs prior to going on pasture.     

Excretion in feces and mucosal persistence of Salmonella ser. Typhimurium in pigs subclinically infected with Oesophagostomum spp

OBJECTIVE: To determine interactions between Oesophagostomum spp and Salmonella ser. Typhimurium in pigs. ANIMALS: 30 healthy 5- to 6-week-old pigs. PROCEDURE: Pigs were allotted to 3 groups (n = 10 pigs/group) and treated as follows: group A was given Oesophagostomum dentatum and O quadrispinulatum; group B was given O dentatum, O quadrispinulatum, and S Typhimurium; and group C was given S Typhimurium only. Pigs in groups A and B were trickle infected with Oesophagostomum spp 3 times weekly throughout the study. After 19 days, groups B and C were inoculated once with S Typhimurium. One pig from each group was euthanatized on the day of Salmonella exposure and 2 and 4 days after Salmonella exposure. The remaining pigs were euthanatized on days 16 and 17 after Salmonella exposure. RESULTS: Pigs with dual infections of nematodes and bacteria (group B) excreted significantly higher amounts of S Typhimurium in feces, compared with nematode-free pigs (group C). In addition, group-B pigs excreted S Typhimurium on more days than pigs in group C. Salmonella Typhimurium was detected in the cecum and colon in the majority of pigs in group B, whereas S Typhimurium was only detected in the colon in pigs in group C. Immunohistochemical examination detected S Typhimurium in 7 of 9 pigs in group B but only 2 of 9 pigs in group C. CONCLUSIONS AND CLINICAL RELEVANCE: Interactions between intestinal nematodes and bacteria may play an important role in the dynamics of S Typhimurium infections.     

Efficacy of moxidectin 0.5% pour-on against swine nematodes

Forty pigs with induced infections of Ascaris suum, Trichuris suis, Metastrongylus spp., Oesophagostomum dentatum and O. quadrispinulatum were assigned to five-dose groups of moxidectin 0.5% pour-on with eight pigs per dose group. The doses were: moxidectin, 0 (vehicle control), 0.75, 1.00, 1.25, and 1.50 mg/kg(-1) body weight. Worm egg counts (EPG) were made from fecal samples collected on Day 2 pretreatment and on Day 14 or 15 post-treatment. Animals were ranked according to the descending order of A. suum egg counts made on Day 2 and blocked in groups of five. Pigs in blocked groups were assigned randomly to each of the five dose groups. Treatment doses were calculated on the basis of weights taken on Day 1 and were administered topically from the neck to the base of the tail. Pigs were housed by pairs in individual pens provided with self-feeders and automatic waterers. Necropsies were performed on equal numbers of pigs from each treatment group on days 14 and 15 post-treatment. Adult and larval worms were collected, identified and counted by standard parasitological techniques. All counts were transformed by Y=log10 (count+1) transformation prior to analysis. A two-way analysis of variance was conducted and treatment effect was tested for significance at the 5% level. Efficacies based on geometric means and optimal doses were as follows: Ascaris suum, 98.3% at 1.25; Metastrongylus spp., 100% at 0.75; Oesophagostomum quadrispinulatum, 100% at 1.50; and Trichuris suis, 93.5% at 0.75. Efficacy for O. dentatum was from 81.3% to 100%; however, the average number of O. dentatum (30) was too small for significance. Two species of lungworms were present, Metastrongylus apri and M. pudendotectus but they were not speciated at necropsy. As reported for several anthelmintics, the efficacy of moxidectin was variable for Trichuris. The highest efficacy was in the 0.75 dose group with six pigs harboring a few or no worms. The lowest efficacy was in the 1.25 group with only two pigs harboring a few or no worms.     

Efficacy of piperazine dihydrochloride against Ascaris suum and Oesophagostomum species in naturally infected pigs

A controlled trial was performed to evaluate the efficacy of piperazine dihydrochloride in a new granular formulation (Ascarex D) against naturally occurring infections with Ascaris suum, Oesophagostomum dentatum and O quadrispinulatum. Treatment effects were estimated on the basis of parasites recoverable from the intestinal contents. Given orally at 200 mg per kg body weight the compound showed an efficacy of 99 to 100 per cent against A suum and the nodular worms. Egg excretion of the respective species was reduced by 98 per cent and 100 per cent six days after treatment. No adverse reactions were observed after the treatment.     

Resistance of Oesophagostomum spp. in pigs to pyrantel citrate

This study was undertaken to determine whether anthelmintic-resistant Oesophagostomum spp. populations occur in Danish swine herds. A controlled field trial on selected sow herds suggested resistance to pyrantel citrate in a herd where faecal egg count depression in response to treatment was only 28.3%. This was confirmed by subsequent experimental infection of pigs where the suspected resistant Oesophagostomum isolate was compared with a susceptible worm isolate. After treatment with the recommended dose of the drug, worm burdens of the suspected isolate were only reduced by 42.6% (p greater than 0.05) in contrast to the susceptible isolate, which was reduced by 94.6% (p less than 0.01). Differential counts of the resistant isolate suggested that O. dentatum might be more resistant than O. quadrispinulatum. The resistant isolate originated from a herd, where the anthelmintic had been administered four times a year over a period of 7 years. The potential for development of anthelmintic resistance in Oesophagostomum spp. under management practices in our country is discussed in the light of the current views on predisposing factors.