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Analyse des stéroïdes dans la salive de cochettes pour identifier des biomarqueurs de la période de réceptivité à l'effet mâle

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Poster présenté par Ghylène Goudet (INRA) et al., aux 51es Journées de la Recherche Porcine, 5 et 6 février 2019

En élevage porcin,la conduite en bandes présente des avantages pour la gestion des animaux et l'organistion de l'élevage. Pour synchroniser les cycles des cochettes et les intégrer dans les bandes, la majorité des éleveurs administre un agoniste de synthèse de la progestérone. Afin de limiter le recours aux traitements hormonaux, notre objectif à long terme est de développer des alternatives à l'utilisation des hormones.  Avant la puberté, les cochettes  atteignent un stade de pré-puberté au cours duquel une exposition au verrat (effet mâle) favoriserait le déclenchement et la synchronisation de la première ovulation.  L'objectif de cette étude est d'améliorer le repérage des femelles à exposer au verrat en identifiant dans la la salive des biomarqueurs de la phase de pré-puberté.

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2019

Analyse des stéroïdes dans la salive de cochettes pour identifier des biomarqueurs de la période de réceptivité à l’effet mâle

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51es Journées de la Recherche Porcine, 5 et 6 février 2019, Paris, p. 43-44, par Ghylène Goudet (INRA) et al., poster

Poster. 

L’élevage porcin conventionnel se caractérise par une conduite en bandes qui présente de nombreux avantages pour la gestion des animaux (inséminations, surveillance des mises-bas, ajustement de la taille des portées, soins aux porcelets), l’organisation de l’élevage (utilisation optimale des bâtiments, nettoyage des locaux entre bandes) et la production de lots de porcelets homogènes pour l’engraissement et l’abattage. La conduite en bandes nécessite la synchronisation des cycles des femelles, réalisée par administration d’agonistes de synthèse de la progestérone chez une majorité d’éleveurs. 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 puberté, les cochettes atteignent une phase de pré-puberté au cours de laquelle une stimulation externe peut déclencher la première ovulation (Prunier, 1989). L’exposition au verrat (appelée effet mâle) pourrait favoriser le déclenchement et la synchronisation de la première ovulation si elle est appliquée pendant cette phase de pré-puberté. Notre objectif est d’améliorer le repérage des femelles à stimuler en identifiant des biomarqueurs de la phase de pré-puberté. Ces biomarqueurs sont recherchés dans la salive des cochettes car les prélèvements de salive sont non-invasifs.

Analysis of steroids in gilt saliva to identify biomarkers of the period of receptivity to the boar effect

Our objective was to develop alternatives to hormonal treatments to synchronize oestrus of gilts. Before puberty, gilts experience a pre-puberty period during which boar exposure could induce and synchronize the first ovulation. To develop practical non-invasive tools to identify this period and 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 x 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. They were then slaughtered to ascertain puberty. 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, six did not show oestrus before slaughter and were considered non-receptive. Salivary steroidome analysis was performed for six receptive and six non-receptive gilts using gas chromatography coupled to tandem mass spectrometry. Four saliva samples per gilt were analysed: 26 days and 11 days before boar introduction (day0-26 and day0-11), the day of boar introduction (day0), 3 days later for receptive gilts (day0+3) or 7 days later for non-receptive gilts (day0+7). Saliva analysis detected 30 steroids. The concentrations of six of them were higher (P < 0.05) in receptive gilts than in non-receptive gilts at day0-26 (progesterone), day0-11 (3520-hexahydroprogesterone, 3520-hexahydroprogesterone, dehydroepiandrosterone, androstenediol) and day0 (35-tetrahydroprogesterone). Their low and variable concentrations in saliva require expensive analysis and limit their use in pig farms. However, progesterone could be an interesting biomarker of the pre-puberty period.

2019

Integrating blood transcriptome and immunity traits to identify markers of immune capacity in pigs

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T. Maroilley et al., 69th Annual meeting of the european federation of animal science (EAAP), Dubrovnik, Croatie, 27-31 août 2018

Understanding individual variability of immune capacity in livestock has become a priority to improve sustainability, with the aim to increase disease resistance and resilience in breeding programs. In this study, 550 60-day-old French Large White pigs vaccinated against Mycoplasma hyopneumoniae (M hyo) were monitored for 55 immunity traits (ITs) measured from blood samples (SUS_FLORA ANR funded project). The ITs included two types of parameters. First, parameters directly measured from blood: complete blood counts, counts of various cell subsets by flow cytometry, serum dosage of anti-M hyo IgG and haptoblobin. Second, parameters measured after in vitro stimulation of total blood: phagocytosis, production of cytokines (IL-1&”6;, IL-8, IL-10, IL-17, TNF&”5;, IFN&”7;) after stimulation by LPS or mitogenic agents. All animals were genotyped with 60K Illumina SNP chips. A subset of 243 piglets was chosen for blood transcriptome analysis using Agilent microarrays. We explored covariations between blood expression profiles and IT levels, and could draw lists of the most correlated genes with each IT. Each list represented candidate blood biomarkers potentially predictive of IT variations. As an example, we found 134 genes associated with phagocytosis capacity and we identified a subset of genes that could significantly predict levels of eight ITs related to phagocytosis by a sPLS approach. This gene subset included CXCR1, CCR1 and TLR2. Few candidate biomarkers were previously shown to be genetically controlled for their transcription in blood by eGWAS. Thus, our results provide new data to decipher the genetic architecture of IT variations. A next step will be to understand how IT variations could reflect individual robustness while facing pathogens, and how blood biomarkers could be used as predictors of immune capacity.

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2018

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.

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2017

Development of an oxidative stress model in weaned piglets highlighting plasma biomarkers specificity to different stress inducers

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Poster.

In order to determine the effects of vaccination, heat stress and dietary antioxidant concentration and investigate some blood parameters as an effective model of oxidative stress, 360 weaned piglets (8.7 ± 1.6 kg) were randomly affected to eight groups in a 2x2x2 factorial design. Vaccination at weaning against PCV2 and porcine influenza, heat stress at days 9-10, 23-24 and 37-38 (37°C over 2×6h periods), or controls, were applied to piglets which were given from weaning to day 14, NRC (2012) levels for vitamin E and selenium or extra supplementation in vitamin E, selenium and Superoxide Dismutase-rich melon pulp concentrate. Piglets were housed in 2 identical rooms, with 3 pens per treatment and 15 piglets per pen. Blood samples were taken from 6 piglets per treatment at days 13 and 40, and plasma haptoglobin, gluthatione peroxidase activity, protein and lipid peroxides were determined. Half-time to haemolyse (T1/2) red blood cells (RBC) exposed to a controlled free radical attack was determined in full blood and RBC concentrate.
Both vaccination and heat stress decreased feed intake and daily gain (p<0.05) from day 14 to 41. Whole blood and RBC T1/2 were increased by antioxidant supplementation (p<0.01) and decreased by heat stress (p<0.05). The antioxidant supplementation also increased gluthatione peroxidase activity (p<0.001). Vaccination induced higher haptoglobin and lipid peroxide concentrations (p<0.001). Plasma biomarkers were therefore specifically responsive to either vaccination or antioxidant supplementation and heat. This model of oxidative stress highlights the need to finely select the biomarkers related to each specific stressor.

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2015

Le métabolome, un moyen pour trouver de nouveaux biomarqueurs ? Des profils métabolomiques pour faire de la prédiction de phénotypes chez le porc en croissance

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Cette étude visait à quantifier le pouvoir prédictif du métabolome sur des phénotypes de production dans la filière porcine. Des données métabolomiques ont été obtenues par Résonance Magnétique Nucléaire à partir d’une prise de sang réalisée pendant la période de croissance de l’animal. Selon le type de phénotype considéré, un spectre métabolomique peut être envisagé comme une source de biomarqueurs intéressants.
Ainsi, le taux de muscle des pièces de la carcasse a pu être correctement prédit à partir des mesures prises sur des prélèvements réalisés à environ 60 kg.

ENG

Metabolomics, a way to find new biomarkers?

This study was aimed at quantifying the predictive power of metabolomic data on production phenotypes in the pig. Metabolomic data
were obtained by Nuclear Magnetic Resonance from one blood sample during the growing period of the animals. Depending on the phenotype, a metabolomic spectrum can be envisioned as a source of suitable biomarkers. In particular, the lean meat percentage was correctly predicted from samples taken at around 60 kg.

2014