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Salivary and urinary metabolome analysis for pre-puberty-related biomarkers identification in porcine

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Gylène Gaudet et al., Animal, 2018, 5 septembre, 11 pages

Estrus synchronization is important for optimal management of gilt reproduction in pig farms. Hormonal treatments, such as synthetic progestogens, are used on a routine basis, but there is a growing demand for non-hormonal alternative breeding tools. Before puberty, gilts exhibit a 'waiting period,' related to the ovarian development and gonadotrophin secretions, during which external stimulations, such as boar exposure, could induce and synchronize first ovulation. Practical non-invasive tools for identification of this period in farms are lacking. During this period, urinary oestrone levels are high, but urine sampling is difficult in group-housed females. The aim of this work was to search for specific biomarkers of the 'waiting period' in saliva and urine. In total, nine 144- to 147-day-old Large White gilts were subjected to trans-abdominal ultrasonography three times a week for 5 weeks until puberty detection (week -5 to week -1 before puberty). Urine and saliva samples were collected for oestrone assay to detect the 'waiting period' and for metabolome analysis using 1H-nuclear magnetic resonance spectroscopy to detect potential biomarkers of the 'waiting period.' Gilts were slaughtered 7 days after puberty detection for puberty confirmation. Results were consistent with ultrasonography data for six gilts. Urine and saliva samples from these six gilts were analyzed. Urinary estrone concentration significantly increased 2 weeks before puberty detection. Metabolome analysis of urine samples allowed the identification of 78 spectral bins, among them, 42 low-molecular-weight metabolites were identified. Metabolome analysis of salivary samples allowed the identification of 59 spectral bins, among them, 23 low-molecular-weight metabolites were detected and 17 were identified. No potential biomarker was identified in urinary samples. In saliva, butyrate and 2HOvalerate, 5.79 ppm (putatively uridine), formate, malonate and propionate could be biomarker candidates to ascertain the pre-puberty period in gilt reproduction. These results confirm that non-invasive salivary samples could allow the identification of the physiological status of the gilts and presumably the optimal time for application of the boar effect. This could contribute to synchronize puberty onset and hence to develop non-hormonal breeding tools.

2018

Mesure des concentrations des stéroïdes dans la salive de cochettes immatures, pré-pubères et pubères

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Ghylène Goudet et al., 49es Journées de la Recherche Porcine, Paris, 31 janvier et 1er février 2017, p. 175-176, poster

L’élevage porcin conventionnel se caractérise par une conduite en bandes qui présente de nombreux avantages techniques (surveillance des mises-bas, ajustement de la taille des portées, gestion des porcelets…) et sanitaires (nettoyage-désinfection des locaux entre bandes). Une majorité d’éleveurs administrent des agonistes de synthèse de la progestérone pour synchroniser les cycles des cochettes de renouvellement et les intégrer dans les bandes (Boulot et al., 2005). Les effets négatifs des résidus hormonaux sur la santé humaine et l’environnement conduisent à mettre en place de nouvelles pratiques d’élevage. Notre objectif à long terme est de développer des alternatives aux traitements hormonaux pour la synchronisation des oestrus des cochettes, notamment lors de l’entrée dans la première bande.
Avant la première ovulation, les cochettes atteignent un stade physiologique de pré-puberté au cours duquel une stimulation externe peut déclencher la première ovulation. L’exposition au verrat (appelée effet mâle) pourrait favoriser le déclenchement et la synchronisation de la puberté si elle est appliquée pendant cette période de pré-puberté (Prunier, 1989). Cette pratique est très peu utilisée en élevage, car le moment optimal et les modalités d’exposition au verrat ne sont pas clairement définis. Notre objectif est de mieux caractériser la phase de pré-puberté et de rechercher des biomarqueurs de cette phase à l’aide de techniques non-invasives.
Pendant la phase de pré-puberté, les concentrations d’oestrone urinaire augmentent (Camous et al., 1985). En conditions d’élevage avec logement en groupe, des prélèvements réguliers d’urine sont difficilement envisageables. Les dosages hormonaux classiques reposent sur des prélèvements sanguins trop invasifs. En revanche, le suivi de biomarqueurs salivaires est non invasif et relativement facile à mettre en place. La recherche de biomarqueurs salivaires de la phase de prépuberté permettra d’améliorer le repérage des femelles à stimuler et de diminuer le nombre de femelles mises à la reproduction alors qu’elles sont impubères.

ENG

Evaluation of steroids concentrations in the saliva of immature, prepubertal and pubertal gilts

Synchronizing oestrus in gilts is an important tool for pig producers to optimize the management of reproduction. Synthetic progestogens are used routinely in pig farms for this purpose, but there is a need for alternative non-hormonal breeding tools. Before puberty, gilts exhibit a “waiting period”, related to ovarian development and gonadotrophin secretions, during which an external stimulation, such as boar exposure, could induce and synchronize the first ovulation. Our aim was to characterize this “waiting period” and search for biomarkers of this period using non-invasive tools. During this period, an increase of urinary oestrone concentration has been observed, but urinary sampling is difficult in group-housed females. Salivary samples are non-invasive and easier to perform. Thus, the aim of this study was to evaluate steroid concentrations in the saliva of immature to pubertal gilts in order to search for biomarkers of the “waiting period”. Trans-abdominal ultrasonographies were performed over a period of 5 weeks on six 140-day-old Large White gilts until puberty detection. Urinary samples were collected for oestrone assay to detect the “waiting period”. Salivary samples were collected for steroidome analysis using gas chromatography coupled with tandem mass spectrometry to detect potential biomarkers of the “waiting period”. Urinary oestrone concentration increased 2 weeks before puberty. Steroidome analysis allowed the quantification of 28 steroids, 13 of them showing significant differences between weeks. Dehydroepiandrosterone concentrations decreased and estradiol-17 concentrations increased significantly just before the “waiting period”. These steroids could be biomarkers of this period. Present results confirm that non-invasive salivary samples could allow the identification of the physiological status of the gilts and the optimal time for application of the boar effect.

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2017

Evaluation of steroid concentrations in the saliva of pre-pubertal gilts for the identification of biomarkers of the pubertal stage of maturity

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Ghylène Goudet et al., ESDAR, 24-26 août 2017, Bern, Suisse, abstract

Estrus synchronization is important for optimal management of gilt reproduction in farms. Synthetic progestagens are used for this purpose, but there is growing demand for non-hormonal alternatives. Before puberty, gilts exhibit a “waiting period”, related to ovarian development and gonadotrophin secretions, during which external stimulations, such as boar exposure, could induce and synchronize first ovulation. Practical non-invasive tools for identification of this period in farms are lacking. During this period, urinary estrone levels are high, but urine sampling is difficult in group-housed females. Our aim was to search for steroidal biomarkers of this “waiting period” from immature to pubertal gilts through saliva monitoring.
Six 140-day-old Large White gilts were subjected to ultrasound puberty diagnosis 3 times a week for 5 weeks until first ovulation. Urine and saliva samples were collected at the same frequency for estrone assay and steroidome analysis respectively. Urinary estrone concentration significantly increased 2 weeks before puberty (detected at 182-192 days). Steroidome analysis quantified 28 steroids in saliva. Significant variations were detected within 2 weeks before puberty for dehydroepiandrosterone (decrease) and estradiol-17&a538; (increase). These steroids could be biomarkers of the “waiting period”.
These results confirm that non-invasive salivary sampling could allow the identification of the physiological status of the gilts and presumably the optimal time for application of the boar effect.

Document réservé Espace Pro, veuillez vous identifier
2017

Exactitude d'un kit de dosage rapide de la progestérone chez la truie

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Despite new applications of ultrasonography, blood progesterone is still a reference diagnostic tool for the assessment of reproductive status in many species, including the sow (2, 5). Techniques based on saliva or fecal samples (3), have been investigated, but only serum semi-quantitative ELISA (1) can be implemented at low costs. This study was designed to validate a new ELISA kit, for rapid field determination of sow progesterone.
PDF icon Exactitude d'un kit de dosage rapide de la progestérone chez la truie
2007