体外 Heather（石南科）提取物对不同发育阶段的捻转血矛线虫和食道口线虫的影响。
In vitro effect of heather (Ericaceae) extracts on different development stages of Teladorsagia circumcincta and Haemonchus contortus.
机构信息
SALUVET Group, Department of Animal Health, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain.
出版信息
Vet Parasitol. 2013 Oct 18;197(1-2):235-43. doi: 10.1016/j.vetpar.2013.05.010. Epub 2013 May 21.
The aim of the present study was to evaluate the in vitro effects of heather (Ericaceae) phenolic extracts on the abomasal nematodes Teladorsagia circumcincta and Haemonchus contortus. Extracts of three heather species (Calluna vulgaris, Erica cinerea, Erica umbellata and a balanced mixture of all three) were tested in vitro on different development stages of T. circumcincta (eggs, infective larvae and adult worms) and H. contortus (eggs and infective larvae) using an egg hatching assay (EHA), a larval exsheathment inhibition assay (LEIA) and an adult motility inhibition assay (AMIA). The egg hatching rate was measured after incubation with heather extracts for 48 h at 25 °C. Ensheathed infective larvae were incubated for 3h at 20 °C with heather extracts. Artificial exsheathment was induced in vitro by adding hypochloride solution to the larval suspension. The progress of exsheathment over time was measured by repeated observations at 10-min (T. circumcincta) and 20-min (H. contortus) intervals for 60 min. Adult T. circumcincta worms were obtained from two donor goats and incubated with the extracts at 37 °C for 3 days in 48-well multiwell plates. Worm motility was measured at 0, 19, 24, 43, 48, 67 and 72 h. The extracts were tested at concentrations of 75, 150, 300, 600 and 1200 μg/mL. Incubation with E. cinerea, E. umbellata and mixed heather extracts had a significant (P<0.01) dose-dependent effect on T. circumcincta egg hatching. H. contortus egg hatching was significantly (P<0.01) inhibited only by the E. cinerea extract. All extracts had a significant (P<0.01) dose-dependent effect on the exsheathment of T. circumcincta and H. contortus infective larvae. The incubation with all heather extracts induced a reduction in adult T. circumcincta motility compared to the control, although significant (P<0.05) differences were only found at the highest concentration (1200 μg/mL). The effect of the mixed extract was significant at all concentrations and significant effects were also observed for C. vulgaris and E. umbellata at 600 μg/mL. These results show anthelmintic properties of heather phenolic extracts against T. circumcincta and H. contort, thus confirming observations from previous in vivo studies.
本研究旨在评估石南（石南科）酚类提取物对皱胃线虫（Teladorsagia circumcincta）和捻转血矛线虫（Haemonchus contortus）的体外作用。使用卵孵化试验（EHA）、幼虫蜕皮抑制试验（LEIA）和成虫运动抑制试验（AMIA），在体外测试了三种石南物种（普通石南、石南艾丽西亚、石南伞状）的提取物（普通石南、石南艾丽西亚、石南伞状）在不同发育阶段的 T. circumcincta（卵、感染性幼虫和成虫）和 H. contortus（卵和感染性幼虫）中的作用。在 25°C 下孵育 48 小时后测量卵的孵化率。用石南提取物孵育包被的感染性幼虫 3 小时，温度为 20°C。通过向幼虫悬浮液中添加次氯酸盐溶液，在体外诱导人工蜕皮。通过在 60 分钟内每隔 10 分钟（T. circumcincta）和 20 分钟（H. contortus）重复观察来测量随时间的蜕皮进展。从两只供体山羊中获得成年 T. circumcincta 蠕虫，并在 37°C 下在 48 孔微孔板中用提取物孵育 3 天。在 0、19、24、43、48、67 和 72 小时测量蠕虫运动。在 75、150、300、600 和 1200μg/mL 浓度下测试提取物。E. cinerea、E. umbellata 和混合石南提取物孵育对 T. circumcincta 卵孵化有显著（P<0.01）的剂量依赖性作用。H. contortus 卵孵化仅被 E. cinerea 提取物显著（P<0.01）抑制。所有提取物对 T. circumcincta 和 H. contortus 感染性幼虫的蜕皮均有显著（P<0.01）的剂量依赖性作用。与对照相比，所有石南提取物孵育均诱导成年 T. circumcincta 运动减少，尽管仅在最高浓度（1200μg/mL）下发现显著差异（P<0.05）。混合提取物的作用在所有浓度下均具有统计学意义，在 600μg/mL 时也观察到 C. vulgaris 和 E. umbellata 的显著作用。这些结果表明石南酚类提取物对 T. circumcincta 和 H. contort 具有驱虫特性，从而证实了先前体内研究的观察结果。
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