Untargeted Multimodal Metabolomics Investigation of the Haemonchus contortus Exsheathment Secretome

Abstract

In nematodes that invade the gastro-intestinal tract of the ruminant, the process of larval exsheathment marks the transition from the free-living to the parasitic stages of these parasites. To investigate the secretome associated with larval exsheathment, a closed in vitro system that effectively reproduces the two basic components of an anaerobic rumen environment (CO2 and 39 °C) was developed to trigger exsheathment in one of the most pathogenic and model gastrointestinal parasitic nematodes, Haemonchus contortus (barber‘s pole worm). This study reports the use of multimodal untargeted metabolomics and lipidomics methodologies to identify the metabolic signatures and compounds secreted during in vitro larval exsheathment in the H. contortus infective third-stage larva (iL3). A combination of statistical and chemoinformatic analyses using three analytical platforms revealed a panel of metabolites detected post exsheathment and associated with amino acids, purines, as well as select organic compounds. The major lipid classes identified by the non-targeted lipidomics method applied were lysophosphatidylglycerols, diglycerides, fatty acyls, glycerophospholipids, and a triglyceride. The identified metabolites may serve as metabolic signatures to improve tractability of parasitic nematodes for characterizing small molecule host–parasite interactions related to pathogenesis, vaccine and drug design, as well as the discovery of metabolic biomarkers.

Analysis of fecal microbiome and metabolome changes in goats with pregnant toxemia

“…The separation and detection of metabolites were performed using the Waters2D UPLC tandem Q Exactive HF high-resolution mass spectrometer. For detailed statistical tests performed during microbiome analysis, refer to the works of Palevich et al [ 37 ].…”

Metabolomic analysis reveals a differential adaptation process of the larval stages of Anisakis simplex to the host environment

“…Our results revealed that specific to L3 are nicotinic acid and branched-chain and aromatic amino acids (BCAA). The recent metabolomics studies showed that BCAA are positively related to longevity common to Dauer in C. elegans and Haemonhus contortus during mitochondrial biogenesis in ensheathment eggs ( Palevich et al, 2022 ). In this study, BCAA were only identified in L3 stage, which like a Dauer stage, uses endogenous energy sources.…”

“…Similarly, parasitic nematode research increasingly relies on metabolomics techniques due to its immense potential applications in studying these organisms effectively. The gastrointestinal tract is a dynamic metabolic and immunologically active ecosystem, and its complete set of metabolites reflects both the enzymatic pathways of host and gut inhabitants and the complex network that connects them ( Palevich et al, 2021 , 2022 ; Whitman et al, 2021 ). The use of metabolomics is particularly suited to understanding nematode metabolism, including the identification of novel drug targets, differences between developmental stages, and mechanisms involved in host-parasite interactions ( Jasmer et al, 2019 ; Molenaars et al, 2021 ).…”

Non-targeted multimodal metabolomics data from ovine rumen fluid fractions

“…Quality control and peak deisotoping analysis were based on our previously published procedures ( 10 ). The details of extraction procedures, chromatographic gradients, and instrument settings have been previously described by Palevich et al ( 11 ). For each metabolomic data set, principal component analysis (PCA) was performed on the log 10 intensities for raw chromatographic peaks to assess similarity and separation of DRF fractions.…”