Department of Veterinary Medicine, University of Cambridge, Cambridge, CB3 0ES, UK.
Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Aberystwyth, SY23 2DA, UK.
Parasit Vectors. 2022 Oct 2;15(1):354. doi: 10.1186/s13071-022-05443-z.
Over the past decade, evidence has emerged of the ability of gastrointestinal (GI) helminth parasites to alter the composition of the host gut microbiome; however, the mechanism(s) underpinning such interactions remain unclear. In the current study, we (i) undertake proteomic analyses of the excretory-secretory products (ESPs), including secreted extracellular vesicles (EVs), of the 'brown stomach worm' Teladorsagia circumcincta, one of the major agents causing parasite gastroenteritis in temperate areas worldwide; (ii) conduct bioinformatic analyses to identify and characterise antimicrobial peptides (AMPs) with putative antimicrobial activity; and (iii) assess the bactericidal and/or bacteriostatic properties of T. circumcincta EVs, and whole and EV-depleted ESPs, using bacterial growth inhibition assays.
Size-exclusion chromatography was applied to the isolation of EVs from whole T. circumcincta ESPs, followed by EV characterisation via nanoparticle tracking analysis and transmission electron microscopy. Proteomic analysis of EVs and EV-depleted ESPs was conducted using liquid chromatography-tandem mass spectrometry, and prediction of putative AMPs was performed using available online tools. The antimicrobial activities of T. circumcincta EVs and of whole and EV-depleted ESPs against Escherichia coli were evaluated using bacterial growth inhibition assays.
Several molecules with putative antimicrobial activity were identified in both EVs and EV-depleted ESPs from adult T. circumcincta. Whilst exposure of E. coli to whole ESPs resulted in a significant reduction of colony-forming units over 3 h, bacterial growth was not reduced following exposure to worm EVs or EV-depleted ESPs.
Our data points towards a bactericidal and/or bacteriostatic function of T. circumcincta ESPs, likely mediated by molecules with antimicrobial activity.
在过去的十年中,有证据表明胃肠道(GI)寄生虫能够改变宿主肠道微生物组的组成;然而,支持这种相互作用的机制尚不清楚。在目前的研究中,我们(i)对“棕色胃虫”(Teladorsagia circumcincta)的排泄-分泌产物(ESP),包括分泌的细胞外囊泡(EVs)进行蛋白质组学分析,这是世界温带地区引起寄生虫性胃肠炎的主要病原体之一;(ii)进行生物信息学分析,以识别和表征具有潜在抗菌活性的抗菌肽(AMPs);(iii)通过细菌生长抑制试验评估 T. circumcincta EVs 以及全 ESP 和 EV 耗尽的 ESP 的杀菌和/或抑菌特性。
采用排阻色谱法从全 T. circumcincta ESPs 中分离 EVs,然后通过纳米颗粒跟踪分析和透射电子显微镜对 EVs 进行特征分析。采用液相色谱-串联质谱法对 EVs 和 EV 耗尽的 ESPs 进行蛋白质组学分析,利用现有在线工具预测潜在的 AMPs。采用细菌生长抑制试验评估 T. circumcincta EVs 以及全 ESP 和 EV 耗尽的 ESPs 对大肠杆菌的抗菌活性。
在成年 T. circumcincta 的 EVs 和 EV 耗尽的 ESPs 中均鉴定出几种具有潜在抗菌活性的分子。虽然全 ESPs 暴露于大肠杆菌会导致 3 小时内菌落形成单位显著减少,但暴露于虫体 EVs 或 EV 耗尽的 ESPs 后,细菌生长并未减少。
我们的数据表明 T. circumcincta ESPs 具有杀菌和/或抑菌功能,可能由具有抗菌活性的分子介导。