Equine; Follicular phase; Luteal phase; Nuclear magnetic resonance spectroscopy; Pregnancy; Seasonal anoestrus; Small Animals; Food Animals; Animal Science and Zoology
Abstract :
[en] Successful reproductive management of domestic mammals depends primarily upon timely identification of oestrous cycle stages. There is a need to develop an alternative non-invasive, welfare-friendly, accurate and reliable method to identify reproductive cycle stages. This is of particular interest for horse breeders, because horses are high-value farm animals that require careful management and individual monitoring. Saliva sampling is non-invasive, painless and welfare-friendly. Thus, we performed a metabolomic analysis of equine saliva during different reproductive stages to identify changes in the salivary metabolome during anoestrus, the oestrous cycle and early gestation. We compared the saliva and plasma metabolomes to investigate the relationship between the two fluids according to the physiological stage. We collected saliva and plasma samples from six mares during seasonal anoestrus, during the follicular phase 3 days, 2 days and 1 day before ovulation and the day when ovulation was detected, during the luteal phase 6 days after ovulation, and during early gestation 18 days after ovulation and insemination. Metabolome analysis was performed by proton-nuclear magnetic resonance spectroscopy. We identified 58 and 51 metabolites in saliva and plasma, respectively. The levels of four metabolites or groups of metabolites in saliva and five metabolites or groups of metabolites in plasma showed significant modifications during the 4 days until ovulation, ie 3 days prior to and on the day of ovulation. The levels of 11 metabolites or groups of metabolites in saliva and 17 metabolites or groups of metabolites in plasma were significantly different between the seasonal anoestrus and the ovarian cyclicity period. The physiological mechanisms involved in the onset of ovarian cyclicity and in ovulation induced modifications of the metabolome both in plasma and saliva. The metabolites whose salivary levels changed during the reproductive cycle could be potential salivary biomarkers to detect the reproductive stage in a welfare friendly production system. In particular, we propose creatine and alanine as candidate salivary biomarkers of ovulation and of the onset of ovarian cyclicity, respectively. However, extensive validation of their reliability is required. Our study contributes to extend to domestic mammals the use of saliva as a non-invasive alternative diagnostic fluid for reproduction in a welfare-friendly production system.
Disciplines :
Animal production & animal husbandry
Author, co-author :
Goudet, Ghylène ; INRAE, CNRS, IFCE, Université de Tours, PRC, 37380, Nouzilly, France. Electronic address: ghylene.goudet@inrae.fr
Beauclercq, Stéphane; BOA, INRAE, Université de Tours, 37380, Nouzilly, France. Electronic address: beauclercq.stephane@courrier.uqam.ca
Douet, Cécile; INRAE, CNRS, IFCE, Université de Tours, PRC, 37380, Nouzilly, France. Electronic address: cecile.douet@inrae.fr
Reigner, Fabrice; PAO, INRAE, 37380, Nouzilly, France. Electronic address: fabrice.reigner@inrae.fr
Deleuze, Stefan ; Université de Liège - ULiège > Fundamental and Applied Research for Animals and Health (FARAH) > FARAH: Médecine vétérinaire comparée
Nadal-Desbarats, Lydie; UMR 1253, iBrain, INSERM, Université de Tours, 37000, Tours, France. Electronic address: lydie.nadal@univ-tours.fr
Language :
English
Title :
Saliva and plasma metabolome changes during anoestrus, the oestrous cycle and early gestation in the mare: A pilot study.
In mammals, ovulation is triggered by systemic modifications linked to secretion of gonadotropins and steroid hormones and local processes such as steroidogenesis, prostaglandin synthesis and proteolysis leading to follicle rupture, a crucial process for successful ovulation [23]. Multiple endocrine, paracrine and autocrine signaling pathways regulate ovulation. In mares, plasma LH concentrations, assayed by radioimmunoassay, peak one day after ovulation [15]. Evaluation of steroid concentrations in plasma of cyclic mares using enzyme immunoassay and radioimmunoassay showed that progesterone concentrations increase from the day of ovulation to 6 days post-ovulation and then decrease, whereas 17beta-oestradiol concentrations reach a peak 2 days before ovulation and then decrease [1,15]. Using GC-MS/MS analysis of the plasma steroidome in mares, we confirmed the decrease in 17beta-oestradiol concentrations before ovulation, and we showed that dehydroepiandrosterone concentrations decreased significantly on the last day before ovulation [4]. In the present study, using 1H NMR analysis of the plasma metabolome, we showed that the levels of serine (proteinogenic amino acid), dimethyl sulfone (organic sulphur compound belonging to sulfones, source of sulphur) and valine (proteinogenic amino acid) decreased before ovulation. Thus, these two steroids and three metabolites could be potential candidate biomarkers for the detection of ovulation. However, the individual data overlapped and the differences were relatively small. Serine, valine and sulphur consumption may increase throughout the ovulatory process initiated by LH, to support the protein synthesis essential for ovulation, as observed in hamsters [24]. Similarly, valine has been linked to potential increase in ovulation rate in ewes [25]. Additionnaly, sulphur plays a critical role in protecting against oxidative stress, which is vital for the quality of ovulation [26]. However, the analysis of these biomarkers relies on invasive and stressful serial blood sampling, highlighting the need for non-invasive, welfare-friendly techniques for the detection of ovulation.This work was supported by INRAE, Institut Fran\u00E7ais du Cheval et de l'Equitation (IFCE) and University of Li\u00E8ge.
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