Short-term responses of meat ewes facing an acute nutritional challenge in early-mid lactation.

Simulating a consequence of a climate change event on feed availability, responses of Mediterranean meat ewes facing an acute undernutritional challenge (CHA; i.e., fed only low nutritional value cereal straw) were evaluated at a sensitive physiological stage (i.e., early suckling). Forty Romane ewes were chosen at early-mid pregnancy (around 2 mo) according to parity (20 primiparous, PRIM; 20 multiparous, MULT); feed efficiency genetic line of their sires (residual feed intake [RFI]; efficient, RFI-, n = 10 per parity; inefficient, RFI+, n = 10 per parity); litter size (i.e., bearing twins, diagnosed by ultrasonography); body weight (BW, kg) and body condition score (BCS) (initial BW and BCS [mean ±â€…SD]: 51.6 ±â€…7.41 kg; 2.5 ±â€…0.20, respectively; representing flock' averages per parity). Effects on dry matter intake (DMI), ewes' BW and BCS, subcutaneous dorsal fat thickness (DFT), energy metabolism (plasma non-esterified fatty acids [NEFA], ß-hydroxybutyrate (ß-OHB), glucose, urea, triiodothyronine [T3]), and lambs' growth (BW and average daily gain [ADG]; g/d) were examined before, during and after CHA. Individuals' profiles of the response-recovery to CHA were described using a piecewise mixed-effects model. The fixed effect of parity and genetic line and the random effect of individual (ewe) were considered. A linear mixed-effects model was fitted to explore the effects on lambs' growth. The 2-d straw-only CHA had significant effects on most of the recorded parameters. Meaningful drops and recoveries were observed on ewes' DMI, BW, and DFT with effect on postchallenge levels. BW, BCS, DFT, or DMI were also affected by parity (MULT > PRIM) but not by genetic line. Plasma NEFA, ß-OHB, glucose, urea, and T3 responded well to CHA with drops in T3, urea, and glucose levels, whereas NEFA and ß-OHB significantly increased after CHA. MULT ewes presented sharper ß-OHB recovery from CHA than PRIM (P ≤ 0.05). With this study, we provide tangible and necessary data for an emerging field of research. Our results give new insights into how such a short and abrupt CHA affects some key zootechnical and physiological parameters, and to what extent the impacts of CHA and the ewes' response-recovery are influenced. It also revealed potential between-individual differences in the adaptive capacities of ewes, which require further exploration.

Genome-wide analyses reveal a strong association between LEPR gene variants and body fat reserves in ewes.

BACKGROUND: Among the adaptive capacities of animals, the management of energetic body reserves (BR) through the BR mobilization and accretion processes (BR dynamics, BRD) has become an increasingly valuable attribute for livestock sustainability, allowing animals to cope with more variable environments. BRD has previously been reported to be heritable in ruminants. In the present work, we conducted genome-wide studies (GWAS) in sheep to determine genetic variants associated with BRD. BR (i.e. levels) and BRD (i.e. changes over time) were obtained through body condition score measurements at eight physiological stages throughout each productive cycle in Romane ewes (n = 1034) and were used as phenotypes for GWAS. After quality controls and imputation, 48,593 single nucleotide polymorphisms (SNP) were included in the GWAS. RESULTS: Among the 23 QTL regions identified, a major QTL associated with BR during pregnancy and lactation was identified on chromosome 1. In this region, several significant SNPs mapped to the leptin receptor gene (LEPR), among which one SNP mapped to the coding sequence. The point mutation induces the p.P1019S substitution in the cytoplasmic domain, close to tyrosine phosphorylation sites. The frequency of the SNP associated with increased BR levels was 32%, and the LEPR genotype explained up to 5% of the variance of the trait. Higher fatness levels in ewes carrying the LEPR p.P1019S mutation were observed all along the productive cycle. CONCLUSIONS: These results provide strong evidences for involvement of LEPR in the regulation of BR in sheep and highlight it as a major candidate gene for improving adaptive capacities by genetic selection.

Variability in Global DNA Methylation Rate Across Tissues and Over Time in Sheep.

Recent studies highlighted the influence of epigenetic marks in the variability of many complex traits, both in plants and animals. These studied focused only on specific sites of the genome having differentially methylated profiles among individuals and/or tissues. In contrast, we recently used the methylation rate of the entire genome as a unique measure considered as a novel quantitative phenotype in sheep. This phenotype named global DNA methylation rate (GDMR), measured by luminometric assay, integrates the methylation level of each CpG dinucleotide within the 6 million of CCGG sites along the ovine genome. GDMR measured in blood previously showed moderate heritability of 0.20 and provided evidence for a genetic determinism. The main objective of the present study was to better characterize the GDMR phenotype in various tissues and investigate its variability in several breeds of sheep reared in the same environment. GDMR was measured on blood samples collected monthly from 59 growing male and female lambs (24 Romane, 23 Blackbelly and 12 Charollais), between birth and 4 months of age. Blood GDMR was on average around 80% and was influenced by the sampling date (p < 0.001), the breed (p = 0.002) and the sex (p = 0.002). In addition, GDMR was determined in 12 somatic (frontal lobe, pituitary gland, heart, lung, sub cutaneous and perirenal adipose tissue, skeletal muscle, liver, spleen, adrenal gland, medulla and cortical kidney) and 6 reproductive tissues (ovary, oviduct, uterus, testis, epididymis and seminal vesicle). GDMR was on average 70% in somatic tissues but marked variation was observed depending on the tissue. The GDMR measured in blood was higher than that measured in other somatic tissues, and is not a good proxy of less accessible tissues. Female reproductive tissues had a 10% higher GDMR than male reproductive tissues. We demonstrated a significant influence of the breed on blood GDMR, certainly reflecting the influence of different genetic backgrounds. The effect of the breed on GDMR may be related to their specific abilities to adapt to and live in different conditions.

Maternal reactivity of ewes at lambing is genetically linked to their behavioural reactivity in an arena test.

In sheep, the bond between the dam and her lambs is established during the first hours of a lamb's life. Genetic variability for behavioural reactivity of ewes assessed in an arena test performed 24 hr after lambing has already been reported. However, there is no evidence that this reactivity represents the ewe's maternal reactivity at lambing in outdoor conditions. The objective of this study was to investigate whether or not the behavioural reactivity of ewes in the arena test is genetically related to their maternal reactivity measured at lambing. A total of 935 Romane ewes were studied. The maternal reactivity of ewes at the outdoor lambing site was recorded in response to a human approach and to the handling of the lambs. Their behavioural reactivity was also recorded 24 hr post-lambing in the arena test that involved a separation from the litter and a human presence. Flight distance, aggressive reaction, time to restore contact with the litter, maternal behaviour scores and vocalizations recorded at the lambing site were heritable (0.12-0.34). All of these behaviours were genetically correlated with the behavioural reactivity in the arena test. The highest genetic correlations (from 0.60 to 0.90) were found amongst maternal behavioural scores, flight distance and high-pitched bleats. In conclusion, behavioural reactivity in the arena test can be used to assess early maternal reactivity in standardized conditions. Phenotyping of ewes' behavioural reactivity with a simplified arena test can be performed for genetic improvement in maternal behaviour in sheep.

A Non-Invasive Millimetre-Wave Radar Sensor for Automated Behavioural Tracking in Precision Farming-Application to Sheep Husbandry.

The automated quantification of the behaviour of freely moving animals is increasingly needed in applied ethology. State-of-the-art approaches often require tags to identify animals, high computational power for data collection and processing, and are sensitive to environmental conditions, which limits their large-scale utilization, for instance in genetic selection programs of animal breeding. Here we introduce a new automated tracking system based on millimetre-wave radars for real time robust and high precision monitoring of untagged animals. In contrast to conventional video tracking systems, radar tracking requires low processing power, is independent on light variations and has more accurate estimations of animal positions due to a lower misdetection rate. To validate our approach, we monitored the movements of 58 sheep in a standard indoor behavioural test used for assessing social motivation. We derived new estimators from the radar data that can be used to improve the behavioural phenotyping of the sheep. We then showed how radars can be used for movement tracking at larger spatial scales, in the field, by adjusting operating frequency and radiated electromagnetic power. Millimetre-wave radars thus hold considerable promises precision farming through high-throughput recording of the behaviour of untagged animals in different types of environments.

Genetic parameters estimates for ewes' behavioural reactivity towards their litter after lambing.

In livestock, improving maternal reactivity towards the litter is an important issue in breeding strategies to promote production and animal welfare. As of yet, no studies have investigated the within-breed genetic variation of maternal reactivity in sheep. The objective of this study was to estimate the genetic parameters of maternal reactivity traits. A total of 1,095 primiparous and 1,441 multiparous Romane ewes were phenotyped 24 hr postlambing using a behavioural test (arena test, AT) over a 10-year experimental period. The test consisted of three successive phases evaluating the ewe's attraction to her litter, reactivity to separation from her litter, and reactivity to a conflict between attraction to her litter and avoidance of a motionless human. The ewes were reared exclusively on rangelands (South of France) and lambed outdoors in the spring. High-pitched bleating and low-pitched bleating in the AT were mostly highly heritable (0.39-0.46). Heritabilities were moderate for proximity to the litter in the presence of a human (0.27) and low for locomotion and vigilance in the AT (0.09-0.15). The measurements of a given behaviour in the three phases of the AT were highly genetically correlated. Few genetic correlations were found between the different behavioural traits in the AT, the highest correlations being between high-pitched bleating and low-pitched bleating (-0.43 to -0.77). In conclusion, our findings demonstrate moderate-to-high heritability for maternal reactivity traits. These traits could be included in genetic selection schemes to enhance maternal attachment provided there is no unfavourable link with other production traits.

Genetic Determinism Exists for the Global DNA Methylation Rate in Sheep.

Recent studies showed that epigenetic marks, including DNA methylation, influence production and adaptive traits in plants and animals. So far, most studies dealing with genetics and epigenetics considered DNA methylation sites independently. However, the genetic basis of the global DNA methylation rate (GDMR) remains unknown. The main objective of the present study was to investigate genetic determinism of GDMR in sheep. The experiment was conducted on 1,047 Romane sheep allocated into 10 half-sib families. After weaning, all the lambs were phenotyped for global GDMR in blood as well as for production and adaptive traits. GDMR was measured by LUminometric Methylation Analysis (LUMA) using a pyrosequencing approach. Association analyses were conducted on some of the lambs (n = 775) genotyped by using the Illumina OvineSNP50 BeadChip. Blood GDMR varied among the animals (average 70.7 ± 6.0%). Female lambs had significantly higher GDMR than male lambs. Inter-individual variability of blood GDMR had an additive genetic component and heritability was moderate (h 2 = 0.20 ± 0.05). No significant genetic correlation was found between GDMR and growth or carcass traits, birthcoat, or social behaviors. Association analyses revealed 28 QTLs associated with blood GDMR. Seven genomic regions on chromosomes 1, 5, 11, 17, 24, and 26 were of most interest due to either high significant associations with GDMR or to the relevance of genes located close to the QTLs. QTL effects were moderate. Genomic regions associated with GDMR harbored several genes not yet described as being involved in DNA methylation, but some are already known to play an active role in gene expression. In addition, some candidate genes, CHD1, NCO3A, KDM8, KAT7, and KAT6A have previously been described to be involved in epigenetic modifications. In conclusion, the results of the present study indicate that blood GDMR in domestic sheep is under polygenic influence and provide new insights into DNA methylation genetic determinism.

Relationships between body reserve dynamics and rearing performances in meat ewes1.

The main objective of this work was to study the relationships between body reserve (BR) dynamics and rearing performance (PERF) traits in ewes from a Romane meat sheep flock managed extensively on "Causse" rangelands in the south of France. Flock records were used to generate data sets covering 14 lambing years (YR). The data set included 1,146 ewes with 2 ages of first lambing (AGE), 3 parities (PAR), and 4 litter sizes (LS). Repeated measurements of the BW and BCS were used as indicators of BR. The ewe PERF traits recorded were indirect measurements for maternal abilities and included prolificacy, litter weight and lamb BW at lambing and weaning, ADG at 1, 2, and 3 mo after lambing, and litter survival from lambing to weaning. The effects of different BW and BCS trajectories (e.g., changes in BW and BCS across the production cycle), previously been characterized in the same animals, on PERF traits were investigated. Such trajectories reflected different profiles at the intraflock level in the dynamics of BR mobilization-accretion cycles. Genetic relationships between BR and PERF traits were assessed. All the fixed variables considered (i.e., YR, AGE, PAR, LS, and SEX ratio of the litter) have significant effects on the PERF traits. Similarly, BW trajectories had an effect on the PERF traits across the 3 PARs studied, particularly during the first cycle (PAR 1). The BCS trajectories only affected prolificacy, lamb BW at birth, and litter survival. Most of the PERF traits considered here showed moderate heritabilities (0.17-0.23) except for prolificacy, the lamb growth rate during the third month and litter survival which showed very low heritabilities. With exception of litter survival and prolificacy, ewe PERF traits were genetically, strongly, and positively correlated with BW whatever the physiological stage. A few weak genetic correlations were found between BCS and PERF traits. As illustrated by BW and BCS changes over time, favorable genetic correlations were found, even if few and moderate, between BR accretion or mobilization and PERF traits, particularly for prolificacy and litter weight at birth. In conclusion, our results show significant relationships between BR dynamics and PERF traits in ewes, which could be considered in future sheep selection programs aiming to improve robustness.

Genetic analysis of robustness in meat sheep through body weight and body condition score changes over time.

Animal robustness may be defined as a complex trait characterizing the ability of an individual to be adapted, productive and healthy under contrasted and fluctuating environmental situations. Such a trait is now considered an essential criterion in order to meet the more ambitious goals of farming sustainability. In ruminants, one of the key mechanisms via which robustness is expressed is the capacity to mobilize or restore body reserves (BR) to cope with the challenges of negative energy balances. The objectives of this work were as follows: 1) to estimate the genetic parameters related to BR dynamics in ewes over successive production cycles and 2) to investigate BR management relationships between different physiological stages. For this, historical individual BW and BCS data from 2,920 phenotyped ewes were used for genetic analysis. The changes in BW (∆BW) and BCS (∆BCS) over time were analyzed. Eight physiological stages were considered to investigate these changes over time: mating, early pregnancy, mid-pregnancy, lambing, early suckling, mid-suckling, weaning, and postweaning. The estimated heritability were low for both ∆BW (h2 = 0.13 to 0.18) and ∆BCS (h2 = 0.04 to 0.16). Moderate to high positive genetic correlations (from 0.48 to 0.91) were obtained between BR mobilization phases and between BR accretion phases. Similarly, moderate to high negative genetic correlations (from -0.36 to -0.75) were estimated between the BR mobilization and accretion periods, suggesting that mechanisms driving BR mobilization and accretion processes were genetically correlated. This is the first study in ruminants that demonstrate that the extent and temporal changes in profiles of BR mobilization and accretion are heritable and genetically linked, indicating that such traits could be considered in genetic programs aimed at improving robustness. Nevertheless, further research is needed for a more comprehensive understanding of BR dynamics, notably by including other physiological parameters (i.e., metabolites and hormones) and additional information on the productive and reproductive life of the ewe.

Identification of QTLs for behavioral reactivity to social separation and humans in sheep using the OvineSNP50 BeadChip.

BACKGROUND: Current trends in sheep farming practices rely on animals with a greater level of behavioral autonomy than before, a phenotype that actively contributes to the sustainability of animal production. Social reactivity and reactivity to humans are relevant behavioral traits in sheep, known for their strong gregariousness and weak tolerance to handling, which have previously been reported with moderate to high heritabilities. To identify loci underlying such behaviors, we performed a genome study in Romane lambs. RESULTS: The experiment was carried out on 934 male and female lambs allocated into 9 half-sib families (average of 103 lambs per family) and reared outside. After weaning, all the lambs were individually exposed to 4 standardized behavioral tests combining social isolation, exposure to humans or handling, confinement and novelty (i.e. arena test, corridor test, isolation box test, shearing test). A broad range of behaviors including vocalizations, locomotion, vigilance and flight distance, as well as the cortisol response to handling, were collected. All lambs were genotyped using the Illumina OvineSNP50 BeadChip. QTL detection was performed by linkage, association and joint linkage and association analyses using the QTLmap software. Five main QTL regions were identified on sheep chromosomes (Ovis Aries Region, OAR) 12, 16, 19, 21 and 23 among many other QTLs with small to moderate effects. The QTLs on OAR12, 16 and 21 showed significant associations with social reactivity. The QTLs on OAR19 and 23 were found to be associated with reactivity to humans. No overlapping QTLs were identified for the different traits measured in the behavioral tests, supporting the hypothesis that different genetic factors influence social reactivity and tolerance to humans. CONCLUSION: The results of this study using ovine SNP data suggest that in domestic sheep the behavioral responses to social separation and exposure to humans are under polygenic influence. The most relevant QTLs reported in the present study contain interesting candidate genes previously described to be associated with various emotional and social behaviors in mammals.

Genetic differences in coping strategies in response to prolonged and repeated restraint in Japanese quail divergently selected for long or short tonic immobility.

Exposure to fearful situations elicits behavioral and Hypothalamic-Pituitary-Adrenal (HPA) axis responses characteristic of the coping response of individual animals to counteract environmental challenges. The aim of this study was to investigate behavioral and corticotropic responses concomitantly following prolonged or repeated restraint stress by placing two genotypes of Japanese quail divergently selected for long (LTI) or short (STI) duration of tonic immobility (TI) in a crush cage. In our study, STI quail exhibited higher corticosterone (CORT) levels than LTI quail in response to prolonged restraint. STI quail struggled sooner and much more than LTI quail, and struggling behavior in STI quail progressively decreased during the course of restraint whereas LTI quail displayed very little struggling behavior in the crush cage. LTI quail are thus more likely to adopt a passive behavior coping strategy upon exposure to threat whereas STI quail behave more as active copers. The corticosterone responses shown by LTI and STI quail under restraint stress suggest that adrenocortical correlates of coping behavior in these genotypes of quail may be different from the coping styles previously described in other species. Repeated restraint slightly decreased CORT responses to stress in all experimental groups, but more markedly in male STI quail, whereas adrenal sensitivity and maximum adrenal corticosterone response capacity did not change in any group. On the other hand, neither behavioral habituation nor sensitization processes occurred in the context of repeated restraint in female and male LTI quail and female STI quail, whereas the decreases observed in some behavioral responses were interpreted to be the result of a habituation process in male STI quail.

Gene array and real time PCR analysis of the adrenal sensitivity to adrenocorticotropic hormone in pig.

BACKGROUND: Variability in hypothalamic-pituitary-adrenal (HPA) axis activity has been shown to be influenced by genetic factors and related to great metabolic differences such as obesity. The aim of this study was to investigate molecular bases of genetic variability of the adrenal sensitivity to ACTH, a major source of variability, in Meishan (MS) and Large White (LW) pigs, MS being reported to exhibit higher basal cortisol levels, response to ACTH and fatness than LW. A pig cDNA microarray was used to identify changes in gene expression in basal conditions and in response to ACTH stimulation. RESULTS: Genotype and/or ACTH affected the expression of 211 genes related to transcription, cell growth/maintenance, signal transduction, cell structure/adhesion/extra cellular matrix and protein kinase/phosphatase activity. No change in the expression of known key regulator proteins of the ACTH signaling pathway or of steroidogenic enzymes was found. However, Mdh2, Sdha, Suclg2, genes involved in the tricarboxylic acid (TCA) pathway, were over-expressed in MS pigs. Higher TCA cycle activity in MS than in LW may thus result in higher steroidogenic activity and thus explain the typically higher cortisol levels in MS compared to LW. Moreover, up-regulation of Star and Ldlr genes in MS and/or in response to ACTH suggest that differences in the adrenal function between MS and LW may also involve mechanisms requisite for cholesterol supply to steroidogenesis. CONCLUSION: The present study provides new potential candidate genes to explain genetic variations in the adrenal sensitivity to ACTH and better understand relationship between HPA axis activity and obesity.