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Steroidome and metabolome analysis in gilt saliva to identify biomarkers of boar effect receptivity

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G. Goudet (INRA) et al., 70th Annual Meeting of the European Federation Animal Science (EAAP), 26-30 août 2019, Ghent, Belgique, p. 489, poster

Our objective was to develop alternatives to hormonal treatments to synchronize oestrus of gilts. Before puberty gilts exhibit a pre-puberty period during which boar exposure could induce and synchronize first ovulation. To develop practical non-invasive tools to identify this period an  improve detection of the gilts to stimulate, we searched for salivary biomarkers of the pre-puberty period. Saliva samples were collected from 30 Large-White × Landrace crossbred gilts from 140 to 175 days of age. Gilts were exposed to a boar twice a day and subjected to oestrus detection from 150 to 175 days of age. Among the 30 gilts, 10 were detected in oestrus 4 to 7 days after introduction of the boar and were considered receptive to the boar effect, 14 were detected in oestrus more than 8 days after boar introduction, 6 did not show oestrus and were considered non-receptive. Saliva samples from 6 receptive and 6 non-receptive gilts were analysed for steroidome using GC-MS/MS and for metabolome using 1H-NMR spectroscopy. Four saliva samples per gilt were analysed: 26 days and 11 days before boar introduction (BI-26 and BI-11), the day of boar introduction (BI), 3 days later for receptive gilts (BI+3) or 7 days later for nonreceptive gilts (BI+7). Data were analysed using repeated measures one-way ANOVA and orthogonal partial least squares discriminant analysis. Thirty steroids and 35 metabolites were detected in gilt saliva. The concentrations of 6 steroids were higher (P<0.05) in receptive gilts than in non-receptive gilts at BI-26, BI-11 and BI. The concentration of 2 metabolites were lower (P<0.05) in receptive gilts than in non-receptive gilts at BI-11. These candidates could be potential salivary biomarkers to detect receptive gilts. However, their low and variable concentrations in saliva require expensive analysis and limit their use in pig farms.

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2019

Evolution of steroid concentrations in saliva from immature to pubertal gilts for the identification of biomarkers of gilts receptivity to boar effect

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Gylène Goudet et al., Livestock science, 2019 volume 228, octobre, p. 5-17

Estrus synchronization is necessary for management of gilt reproduction in pig farms. It is usually achieved by using synthetic progestagens, but there is increasing demand for non-hormonal alternative tools with the prospect of sustainability of livestock production. Moreover, in organic farms, synthetic hormones are not allowed. Before reaching puberty, gilts exhibit a “waiting period” during which external stimulations, such as boar exposure, could trigger and synchronize the first ovulation. However, practical non-invasive tools for detection of the “waiting period” in pig farms are lacking. During this period, estrone levels in urine are high, but urine sampling is difficult in group-housed females. Our objective was to identify among steroids potential biomarkers of this “waiting period” through saliva monitoring from immature to pubertal gilts using gas chromatography coupled to tandem mass spectrometry.

Starting between 144 to 147 days of age, six Large White gilts were submitted to ultrasound puberty diagnosis 3 times a week until first ovulation. Urine and saliva samples were collected to analyze weekly estrone and steroidome respectively, until puberty. Urinary estrone concentration significantly increased 2 weeks before first ovulation occurring between 182 and 192 days of age. The period with increasing estrone levels was considered as the “waiting period”. Steroidome analysis allowed identifying and quantifying 28 steroids in 500 µl of gilts saliva. Significant decrease of dehydroepiandrosterone and significant increase of 5α-dihydroprogesterone and 17β-estradiol were detected 2 weeks before puberty, suggesting that these steroids could be potential biomarkers of the “waiting period”.

These results show that painless sampling of saliva could be a non-invasive welfare-friendly tool for the identification of the physiological hormonal status of the gilts and possibly the optimal time for application of the boar effect, a solution to synchronize puberty without exogenous hormones.

2019

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

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Ghylène Goudet et al., Animal, 2019, volume 13, n° 4, avril, p. 760-770

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.

https://www.cambridge.org/core/services/aop-cambridge-core/content/view/FA6DFF10CB490FA7F83C89B8F3696779/S1751731118002161a.pdf/salivary_and_urinary_metabolome_analysis_for_prepubertyrelated_biomarkers_identification_in_porcine.pdf

2019

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.

https://www.cambridge.org/core/services/aop-cambridge-core/content/view/FA6DFF10CB490FA7F83C89B8F3696779/S1751731118002161a.pdf/salivary_and_urinary_metabolome_analysis_for_prepubertyrelated_biomarkers_identification_in_porcine.pdf

2018

L'échographie, un outil fiable pour détecter la puberté chez la truie

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2006

Conditions de mise en oeuvre et intérêt du contrôle échographique de puberté en élevage porcin

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L’intérêt du contrôle échographique de puberté est évalué pour la 1ère fois dans des conditions d’élevage et avec du matériel de qualité standard. Les mesures ont été réalisées par voie externe, sur 63 cochettes âgées de 180 à 200 jours. L’exactitude des contrôles effectué, avec un appareil portable équipé d’une sonde linéaire, est comparable à celle obtenue avec des appareils de qualité médicale.
PDF icon Conditions de mise en oeuvre et intérêt du contrôle échographique de puberté en élevage porcin
2006

Ultrasonographic field evaluation of puberty using portable standard equipment

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2006

Conditions de mise en oeuvre et intérêt du contrôle échographique de puberté en élevage porcin

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L’intérêt du contrôle échographique de puberté est évalué pour la première fois dans des conditions d’élevage et avec
PDF icon Conditions de mise en oeuvre et intérêt du contrôle échographique de puberté en élevage porcin
2006

Facteurs de variation de l’âge à la puberté, du taux d’ovulation et de la survie embryonnaire précoce chez la cochette : type génétique, poids vif, saison et consanguinité

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Les paramètres de reproduction de truies nullipares Large White, Large White hyperprolifique et Meishan utilisées comme donneuses de jeunes embryons (jour 5 à 11 de gestation) produits à l’élevage expérimental de la station de Physiologie de la Reproduction de Nouzilly ont été analysés entre octobre 1994 et octobre 1996; ils concernent l’âge à la puberté, le taux d’ovulation, le nombre d’embryons et la survie embryonnaire très précoce; ils ont été comparés à ceux publiés antérieurement pour le même troupeau.
PDF icon Facteurs de variation de l’âge à la puberté, du taux d’ovulation et de la survie embryonnaire précoce chez la cochette : type génétique, poids vif, saison et consanguinité
1997