Waghorn Ts, Knight Js, Leathwick Dm
a AgResearch Grasslands , Private Bag 11008, Palmerston North 4442 , New Zealand.
N Z Vet J. 2014 May;62(3):152-9. doi: 10.1080/00480169.2013.871193. Epub 2014 Feb 5.
To determine the distribution of the three common Trichostrongylus spp. infecting sheep and their resistance status on farms throughout New Zealand, using PCR.
Cultures were prepared from faecal samples from 70 farms while conducting faecal egg count reduction tests (FECRT) in lambs between 2010 and 2012. Trichostrongylus-type infective stage larvae (L3) were recovered from cultures, derived from untreated control and albendazole-, levamisole- and ivermectin-treated groups of lambs on each of the farms involved, and these were identified to species using PCR analysis of the second internal transcribed spacer region of ribosomal DNA. The species composition of the larvae present in cultures from the untreated control groups was examined across all farms to assess any potential differences in geographical distribution. In addition, the species composition of larvae cultured from the untreated and anthelmintic-treated lamb groups were compared to determine which species exhibited resistance to each of the anthelmintics used in the FECRT.
Of 67 farms with Trichostrongylus spp. present, 42 (63%) cultures from the untreated control groups contained all three Trichostrongylus spp. and no significant geographical patterns in their distribution were detected. Seven samples contained only one species. Irrespective of the anthelmintic efficacy levels, Trichostrongylus colubriformis dominated cultures prepared from lambs following treatment with albendazole (99.1 (95%CI = 97-100)% of larvae) or levamisole (81.6 (95%CI = 75.3-87.9)% of larvae), indicating the presence of widespread resistance in this species. In cultures prepared from levamisole-treated lambs, small numbers of T. axei larvae were also frequently present (5.4 (95% CI = 1.3-12.4)% of larvae). Resistance to ivermectin was not found in any of the three Trichostrongylus spp. after PCR identification. Although larvae were identified, based on length, as being Trichostrongylus spp., for 24 of the 48 samples cultured following treatment with ivermectin, 100% of the larvae present were identified as Teladorsagia circumcincta.
As in previous surveys, all three Trichostrongylus spp. were common throughout New Zealand and no geographical patterns were detected in the current study. On all farms where resistance to albendazole and/or levamisole was indicated (i.e. efficacy <95%), the species identified as being resistant was T. colubriformis. Even where efficacies were >95%, T. colubriformis still tended to dominate in post-treatment cultures. While this could reflect a lower susceptibility of T. colubriformis to these anthelmintics, it seems more likely to indicate the presence of resistant genotypes in these populations. Similarly, T. axei also tended to be present after treatment with levamisole, which likely reflects a known lower susceptibility of this species to these anthelmintics.
利用聚合酶链反应(PCR)确定感染绵羊的三种常见毛圆线虫的分布情况及其在新西兰各农场的抗药状态。
在2010年至2012年期间对羔羊进行粪便虫卵计数减少试验(FECRT)时,从70个农场的粪便样本中制备培养物。从培养物中回收毛圆线虫型感染性幼虫(L3),这些培养物来自所涉各农场未经处理的对照组以及经阿苯达唑、左旋咪唑和伊维菌素处理的羔羊组,并利用核糖体DNA第二内部转录间隔区的PCR分析将其鉴定到种。对所有农场未经处理的对照组培养物中存在的幼虫种类组成进行检查,以评估地理分布上的任何潜在差异。此外,比较未经处理和经驱虫药处理的羔羊组培养的幼虫种类组成,以确定哪些种类对FECRT中使用的每种驱虫药具有抗性。
在存在毛圆线虫的67个农场中,未经处理的对照组的42个(63%)培养物含有所有三种毛圆线虫,未检测到其分布有明显的地理模式。7个样本仅含有一个种类。无论驱虫药的疗效水平如何,在经阿苯达唑(99.1(95%可信区间=97 - 100)%的幼虫)或左旋咪唑(81.6(95%可信区间=75.3 - 87.9)%的幼虫)处理的羔羊制备的培养物中,蛇形毛圆线虫占主导,表明该种类存在广泛抗性。在经左旋咪唑处理的羔羊制备的培养物中,也经常出现少量的奥氏毛圆线虫幼虫(5.4(95%可信区间=1.3 - 12.4)%的幼虫)。PCR鉴定后,在三种毛圆线虫中均未发现对伊维菌素有抗性的情况。尽管根据长度将幼虫鉴定为毛圆线虫属,但在经伊维菌素处理后培养的48个样本中有24个样本中,所存在的100%的幼虫被鉴定为环纹奥斯特线虫。
与之前的调查一样,在新西兰各地,所有三种毛圆线虫都很常见,本研究未检测到地理模式。在所有显示对阿苯达唑和/或左旋咪唑有抗性(即疗效<95%)的农场中,被鉴定为有抗性的种类是蛇形毛圆线虫。即使疗效>95%,蛇形毛圆线虫在处理后的培养物中仍趋于占主导。虽然这可能反映出蛇形毛圆线虫对这些驱虫药的敏感性较低,但似乎更有可能表明这些种群中存在抗性基因型。同样,在经左旋咪唑处理后也往往会出现奥氏毛圆线虫,这可能反映出该种类对这些驱虫药已知的较低敏感性。