Department of Natural Resources, Agricultural Research Organization, The Volcani Center, Bet Dagan, 50250, Israel.
Department of Entomology and Nematology, Agricultural Research Organization, The Volcani Center, Bet Dagan, 50250, Israel.
Exp Parasitol. 2020 Feb;209:107811. doi: 10.1016/j.exppara.2019.107811. Epub 2019 Dec 3.
As gastro-intestinal nematodes (GINs) become increasingly resistant to chemical anthelmintics, and because consumers scrutinize chemical residues in animal products, the use of herbal anthelmintics and in particular, phenolic compounds, has become attractive. Most life stages of GINs cannot be grown in the lab as they are obligatory parasites, which limits our understanding of the effects of phenolic compounds on their parasitic stages of life. We hypothesized that a species phylogenetically close to GINs and grown in vitro, the insect-parasitic nematode Heterorhabditis bacteriophora (Rhabditida; Heterorhabditiade), when fed with Photorhabdus luminescens exposed to plant phenolics, can serve, as proxy for strongyles, in assessing the anthelmintic effects of phenolic compounds. We compared the development of H. bacteriophora infective juveniles (IJ) and the exsheathment rate of L3 larvae of the strongyle Teladorsagia circumcincta and Trichostrongylus colubriformis when exposed to catechin, rutin, chlorogenic and gallic acids, and myricetin. Gallic acid had the highest impact in terms of IJ mortality but the highest impairment of IJ development to adulthood was imposed by myricetin. The studied compounds were not lethal to GINs stricto sensu but we consider that the practical implications of total exsheathment inhibition and mortality on GIN populations are similar. Catechin and rutin had similar effects on rhabditid and strongyles: they imposed ca. 90% lethality of IJs at concentrations higher than 1200 ppm and the remaining live IJs did not develop further, and they also totally inhibited strongyle L3 exsheathment in a dose-response fashion. Gallic acid was 100% lethal to IJs exposed above 300 ppm and chlorogenic acid caused 87% mortality above 1200 ppm, with no development for the surviving IJs but for all lower concentrations, all the IJs developed to adult stages. Likewise, gallic and chlorogenic acids did not affect the exsheatment of GIN L3 larvae. Therefore, a discrepancy between the effects of gallic and chlorogenic acids on the development of rhabditid IJs and exsheathment of GIN L3 larvae was found only when they were exposed to high concentrations. A dose-response of IJ lethality to myricetin was found, with no IJ development between 150 and 2400 ppm; but contrary to the other compounds, myricetin also impaired IJ development of IJs above 10 ppm in a dose-response manner and showed dose-responses in the L3 exsheathment. Apart for the high rates of lethality imposed on IJs by gallic and chlorogenic acids at high concentration, these results suggest that H. bacteriophora fed P. luminescens exposed to phenolics shows potential to serve as model in studies of the anthelmintic effects of phenolics in GIN.
随着胃肠道线虫(GINs)对化学驱虫剂的抗药性越来越强,而且由于消费者对动物产品中的化学残留物进行了严格审查,因此使用草药驱虫剂,特别是酚类化合物,变得越来越有吸引力。GINs 的大多数生命阶段都不能在实验室中生长,因为它们是专性寄生虫,这限制了我们对酚类化合物对其寄生生命阶段的影响的理解。我们假设,一种与 GINs 亲缘关系密切且在体外生长的昆虫寄生线虫——嗜虫异小杆线虫(Rhabditida;异小杆线虫科),当用暴露于植物酚类化合物的发光光杆菌喂养时,可以作为马副蛔虫的替代品,用于评估酚类化合物的驱虫作用。我们比较了感染性幼虫(IJ)的发育和强捻线虫 Teladorsagia circumcincta 和 Trichostrongylus colubriformis 的 L3 幼虫的脱鞘率,当暴露于儿茶素、芦丁、绿原酸和没食子酸以及杨梅素时。没食子酸在 IJ 死亡率方面的影响最高,但杨梅素对 IJ 发育至成虫的影响最大。研究中的化合物对 GINs 严格意义上没有致死作用,但我们认为完全抑制脱鞘和死亡率对 GIN 种群的实际影响是相似的。儿茶素和芦丁对 Rhabditid 和强捻线虫有相似的影响:它们在高于 1200ppm 的浓度下对 IJ 施加约 90%的致死率,而剩余的活 IJ 不会进一步发育,并且它们还以剂量反应方式完全抑制强捻线虫 L3 的脱鞘。没食子酸在高于 300ppm 的浓度下对 IJ 具有 100%的致死作用,绿原酸在高于 1200ppm 的浓度下导致 87%的死亡率,而存活的 IJ 则没有进一步发育,但对于所有较低的浓度,所有 IJ 都发育成成虫阶段。同样,没食子酸和绿原酸也不影响 GIN L3 幼虫的脱鞘。因此,只有当它们暴露于高浓度时,才会发现没食子酸和绿原酸对 Rhabditid IJ 发育和 GIN L3 幼虫脱鞘的影响之间存在差异。发现了 IJ 致死率对杨梅素的剂量反应,在 150 到 2400ppm 之间没有 IJ 发育;但与其他化合物不同的是,杨梅素在高于 10ppm 的浓度下也会损害 IJ 的发育,并且在 IJ 脱鞘方面也表现出剂量反应。除了高浓度的没食子酸和绿原酸对 IJ 造成的高死亡率外,这些结果表明,用暴露于酚类化合物的发光光杆菌喂养的嗜虫异小杆线虫具有作为研究酚类化合物对 GIN 驱虫作用的模型的潜力。