Sandrine Freret

Expression of adipokine and lipid metabolism genes in adipose tissue of dairy cows differing in a female fertility quantitative trait locus

We have previously characterized 2 haplotypes (Fertil+ and Fertil-) of Holstein dairy cows differing in 1 female fertility quantitative trait locus (QTL) located on chromosome 3 (QTL-Fert-F-BTA3) between positions 9.8 and 13.5 cM. This QTL is composed of 124 genes, some of them being involved in metabolism or reproduction. Primiparous Fertil+ and Fertil- cows exhibited 69 and 39% pregnancy rate at first service, respectively. A difference in plasma nonesterified fatty acid concentrations observed between both haplotypes might indicate a difference in adipose tissue mobilization. We compared adipose tissue gene expression in Fertil+ and Fertil- cows during their second lactation, at 2 physiological stages, implying either intense lipid mobilization (1 wk postpartum) or fat storage (5 mo of gestation). We investigated by reverse-transcription quantitative PCR the mRNA gene expression of 5 positional candidate genes located in the QTL-Fert-F-BTA3, as well as 18 other functional candidate genes encoding proteins involved in lipid metabolism and several adipokines. Among them, genes involved in either lipolysis or lipogenesis were chosen as controls because they were previously described in dairy cow adipose tissue. A hierarchical clustering was performed to group genes according to their expression pattern, allowing 2 clusters to be determined. Cluster 1 was composed of genes that were overexpressed during mobilization (ADIPOQ, ADIPOR2, LIPE, FABP4, PLIN1, RARRES, LEPR, and CPT1A) and cluster 2 of genes overexpressed during reconstitution of body reserves (ACACA, FASN, and SCD). Genes belonging to cluster 1 (LIPE, FABP4, PLIN1, and CPT1A) are known to be involved in lipolysis and fatty acid oxidation, and genes belonging to cluster 2 (ACACA, FASN, and SCD) are known to be involved in fatty acid synthesis. The expression of 5 genes from cluster 1 was correlated to plasma nonesterified fatty acid levels and thus to mobilization of body reserves in dairy cows (ADIPOQ, ADIPOR2, LIPE, PLIN1, and FABP4). During the mobilization stage, none of the positional candidate genes belonging to QTL-Fert-F-BTA3 (ADAR, MTX1, SHC1, SPTA1, and PAQR6) showed a difference in expression between the 2 haplotypes. Interestingly, ADIPOQ and ADIPOR2 were the only genes showing a significant mRNA overexpression in Fertil- cows at the mobilization stage. Further studies focusing on plasma adiponectin level and adipokine actions on the ovary are needed to investigate its potential role in dairy cow fertility.

Effect of a long chain n-3 PUFA-enriched diet on production and reproduction variables in Holstein dairy cows.

The objective of this study was to determine the effect of a rumen-protected fish oil supplement on the production and reproduction variables in postpartum dairy cows. Holstein cows (n=46) were given a basal total mixed diet plus one PUFA supplement: n-3 (n-3; protected fish oil; 1% dry matter intake (DMI); n=23) or control (n-6; toasted soybeans; 1.8% DMI; n=23), in a switchback design over two consecutive lactations. Supplements were added to the diet between calving and 2 months after calving to assess the effect on growth and maturation of ovarian follicles from which ovulation occurred around the day of insemination. Body weight (BW), milk yield (MY) and composition, dry matter intake (DMI), energy balance (EB), subcutaneous fat thickness, plasma fatty acid composition, plasma nonesterified fatty acids (NEFA), glucose and urea concentrations, follicular activity, embryo mortalities and fertility (conception rate after first AI, AI1) were assessed. BW, MY, DMI, plasma NEFA, glucose and urea were unaffected by the diet. There was a trend of an increased number of large follicles (diameter≥10mm) with the n-3 dietary supplementation (P=0.06) and a decrease in infertility or early embryo mortality rate 21 days after AI, 13.5% in the n-3 compared with 38.8% in the n-6 group (P=0.09), with no effect on the conception rate at 35d or 90d after AI1. These data suggest that the effect seen on ovarian variables is not associated with an effect on production and metabolic variables and is specific to n-3 PUFA supplementation. Further studies are necessary to determine whether DHA or EPA enhances fertility in lactating dairy cattle.

Data-derived reference profiles with corepresentation of progesterone, estradiol, LH, and FSH dynamics during the bovine estrous cycle

Subfertility in cows is often associated with alterations in the hormonal patterns involved in the regulation of the estrous cycle. Reference profiles are needed to ground modeling projects aimed at describing these alterations and to develop tools for detecting abnormal dynamics. Various schematic views of LH, FSH, progesterone (P4) and estradiol (E2) patterns have been published but with no clear indication of the extent to which they are derived from real data. The objective of this study was to generate standard profiles for the main reproductive hormones that can be proposed as reliable references to represent the normal dynamics of these hormones over the estrous cycle. A database of hormonal profiles was compiled with 40, 23, 33, and 34 profiles for LH, FSH, E2, and P4, respectively, derived from publications in which changes over time of at least three of these four hormones, including LH, were reported. These profiles were digitalized and standardized over the time throughout the estrous cycle, considering the interval between two successive LH surges to be 21 days. After this standardization on the x-axis, a transformation on the y-axis was performed to center the profiles around their common dynamics. For each hormone, the reference profile was then considered to be the median of the adjusted profiles. Quartiles were reported to account for the time evolution of the variability around each reference profile. The reference profiles obtained showed that the procedure used was satisfactory for extracting the overall changes over time of LH, P4, and E2. Results were less satisfactory for FSH, because of a higher variability observed between the original profiles in our database. The corepresentation of the reference profiles, i.e., when depicted together on the same scale, emphasizes the interplay between these hormones more precisely than most of the schematic views available in literature. These data-derived profiles can be considered to be generic and useful for benchmarking the normal dynamics of gonadotrophins and steroid hormones over the estrous cycle in cow.

A longitudinal study of the Mycobacterium avium subspecies paratuberculosis infection status in young goats and their mothers.

The dynamics between Mycobacterium avium subspecies paratuberculosis (MAP) infection and the immune response of goats naturally exposed to MAP were studied in a herd where the clinical expression of paratuberculosis had been observed. Four generations of goats were observed over a 33-month period: mothers of three different generations (G1, G2, G3) and their daughters, generation 4 (G4). A MAP infection status was defined according to the combined results of an IFN-γ assay, antibody response, faecal culture and post-mortem examination. Goats were defined as non-infected (NI), infected and non-shedder (INS), infected and shedder (IS) or atypical (A). Twenty-nine percent of goats were NI, 66% were infected and either shedding (14%) or not shedding (52%) MAP, and 5% were atypical. IFN-γ responses were detected first, followed by faecal shedding and antibody responses. The results showed that in goats naturally exposed to MAP, IFN-γ responses were regularly detected earlier in non-shedders than in young infected shedder goats and were stronger in shedder than in non-shedder goats. They were also higher in the mother goats than in their daughters. Goats shedding MAP or with positive antibody response at the beginning of their pregnancy are more likely to have an infected daughter positive to an IFN-γ assay by the age of 15 months.

Serum metabolome analysis in doe kids for the identification of biomarkers of sexual precocity

Success of first breeding is a major concern for goat breeders, since failure to fertilize does increases unproductive time and breeding costs. Fertility rates after artificial insemination of young goats are highly variable and rather low. Breeders generally breed does that are older than 5 months and weight more than 32 kg. However, sexual precocity is highly variable between does. Up to now, there is no known biomarker for sexual precocity. A better characterization of the pubertal stage of maturity could help optimizing time for first breeding. Our objective was to analyze the serum metabolome of doe kids, just before the first breeding, in order to characterize the pubertal stage of maturity and identify biomarkers of sexual precocity. Weekly blood sampling was performed on twenty 6-to 7-month-old does born in February for 5 weeks before their first contact with bucks in September. Progesterone assays and metabolome analysis using 1H Nuclear Magnetic Resonance Spectroscopy were performed on the serum samples. No spontaneous ovulatory cycle was observed before breeding based on progesterone assays. All does had reached the pubertal stage of maturity at breeding since all got pregnant. Metabolome analysis allowed the identification of 109 spectral bins in sera. Between week 1 and 5 preceding buck introduction, 32 buckets showed significant variations (t-test, p < 0.05): i.e. inosine, formate, lactate and creatinine decreased, while threonine, tryptophan, isoleucine and trimethylamine oxide significantly increased. Metabolites with significant variations between the 5 considered weeks could be biomarkers of sexual precocity; studies are in progress to identify them.