Untargeted metabolomic profiling of accessory sex gland fluid from Morada Nova rams

Abstract

The present study was conducted to characterize the metabolome of accessory gland fluid (AGF) of locally adapted Morada Nova rams, raised in the Brazilian Northeast. AGF was collected by an artificial vagina from five vasectomized rams. Metabolites were identified by gas chromatography-mass spectrometry (GC/MS) and high-performance liquid chromatography-mass spectrometry (LC/MS), with the support of Human Metabolome Database, PubChem, LIPID Metabolites, Pathways Strategy databases, and MetaboAnalyst platforms. There were 182 and 190 metabolites detected by GC/MS and LC/MS, respectively, with an overlap of one molecule. Lipids and lipid-like molecules were the most abundant class of metabolites in the ram AGF (127 compounds), followed by amino acids, peptides, and analogs(103 metabolites). Considering all GC/MS and LC/MS, fructose, glycerol, citric acid, D-mannitol, D-glucose, and L-(+)-lactic acid were the most abundant single metabolites present in the ram AGF. Meaningful pathways associated with AGF metabolites included glycine, serine and threonine metabolism; pantothenate and CoA biosynthesis; galactose metabolism; glutamate metabolism and phenylalanine metabolism, and so forth. In conclusion, the combined use of LC/MS and GC/MS was essential for getting a holistic view of the compounds embedded in the ram AGF. Chemical analysis of the accessory sex gland secretion is relevant for understanding sperm function and fertilization. K E Y W O R D S accessory sex gland, GC/MS, LC/MS, metabolome, ovine 410 | SOUSA ET AL.

Metabolomics analysis of buck semen cryopreserved with trehalose

“…In this study, the trehalose had a beneficial synergistic effect with low- Frontiers in Genetics frontiersin.org concentration glycerol, which can significantly increase buck sperm total motility and progressive motility after thawing and preserve sperm structural integrity, which was consistent with the findings of previous studies Akhtarshenas et al, 2018;Öztürk et al, 2020). However, optimum cryopreservation requires maintaining metabolic function, and metabolomic approaches allow the study of cellular metabolites, which are thought to be closer to actual phenotypes than proteomes or genomes (Sousa et al, 2020).…”

“…In this study, the trehalose had a beneficial synergistic effect with low-concentration glycerol, which can significantly increase buck sperm total motility and progressive motility after thawing and preserve sperm structural integrity, which was consistent with the findings of previous studies ( Bucak et al, 2007 ; Akhtarshenas et al, 2018 ; Öztürk et al, 2020 ). However, optimum cryopreservation requires maintaining metabolic function, and metabolomic approaches allow the study of cellular metabolites, which are thought to be closer to actual phenotypes than proteomes or genomes ( Sousa et al, 2020 ). Therefore, this study used GC–MS-based untargeted metabolomic analysis technology to observe the effect of trehalose on the metabolome of ram frozen semen, which will help us understand the mechanism of the cryoprotective effect of trehalose on sperm ( Panner Selvam et al, 2019 ).…”

“…In addition, some metabolites, such as L-isoleucine, L-leucine, L-threonine, and dihydroxyacetone may serve as potential biomarkers for assessing the quality of buck sperm after thawing. ( Sousa et al, 2020 ).…”

Nuclear Magnetic Resonance (NMR) Metabolomics: Current Applications in Equine Health Assessment

“…The aim was to identify new molecules that could be used to diagnose infertile subjects with increased sensitivity and specificity [58]. Studies have been performed on seminal plasma in humans [58][59][60] and in different animal species [61][62][63][64]. Three of them [51][52][53] have focused on equine seminal plasma, providing interesting insights into the metabolites contained in this biofluid.…”

Exploring the metabolome of seminal plasma in two different horse types: Light versus draft stallions

“…This approach would also permit the identification of new biomarkers that could be used to diagnose infertile subjects with increased sensitivity and specificity (Engel et al, 2019 ). Studies have been performed on seminal plasma from different species, including bull (Kosior et al, 2020 ; Kumar et al, 2015 ), ram (Sousa et al, 2020 ), stallion (Magistrini et al, 2000 ), boar (Mateo‐Otero et al, 2021 ) and man (Engel et al, 2019 ; Paiva et al, 2015 ; Wang et al, 2019 ), providing interesting insights into the metabolites contained in seminal plasma and its associations with sperm functions and fertility. Metabolomics applied to animal breeding aims to improve prediction of the breeding values of the animals to cope with traditional and new objectives of the selection programs (Kalaiselvi et al, 2019 ).…”