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The search for the genetic determinism of prolificacy variability in sheep has evidenced several major mutations in genes playing a crucial role in the control of ovulation rate. In the Noire du Velay (NV) sheep population, a recent genetic study has evidenced the segregation of such a mutation named FecLL. However, based on litter size (LS) records of FecLL non-carrier ewes, the segregation of a second prolificacy major mutation was suspected in this population. In order to identify this mutation, we have combined case/control genome-wide association study with ovine 50k SNP chip genotyping, whole genome sequencing and functional analyses. A new single nucleotide polymorphism (OARX:50977717T>A, NC_019484) located on the X chromosome upstream of the BMP15 gene was evidenced highly associated with the prolificacy variability (P =1.93E-11). The variant allele was called FecXN and shown to segregate also in the Blanche du Massif Central (BMC) sheep population. In both NV and BMC, the FecXN allele frequency was estimated close to 0.10, and its effect on LS was estimated at +0.20 lamb per lambing at heterozygous state. Homozygous FecXN carrier ewes were fertile with increased prolificacy in contrast to numerous mutations affecting BMP15. At the molecular level, FecXN was shown to decrease BMP15 promoter activity and to impact BMP15 expression in oocyte. This regulatory action was proposed as the causal mechanism for the FecXN mutation to control ovulation rate and prolificacy in sheep.Author SummaryIn the genetic etiology of women infertility syndromes, a focus was done on the oocyte-expressed BMP15 and GDF9 genes harboring several mutations associated with ovarian dysfunctions. In sheep also, mutations in these two genes are known to affect the ovarian function leading to sterility or, on the opposite, increasing ovulation rate and litter size constituting the prolificacy trait genetically selected in this species. Through a genome-wide association study with the prolificacy phenotype conducted in the French Noire du Velay sheep breed, we describe a novel mutation located in the regulatory region upstream of the BMP15 gene on the X chromosome. This mutation increases litter size by +0.2 lamb per lambing at the heterozygous state, possibly through an inhibition of BMP15 expression within the oocyte. Our findings suggest a novel kind of BMP15 variant responsible for high prolificacy, in contrast to all other BMP15 variants described so far in the coding sequence.

International audience; The effects of maternal nutrition on offspring phenotypes have been mainly documented over the past years in mammals, and are now studied in poultry as well. In the present study, we investigated the effects of a reduced level of dietary Methionine (Met) on laying performances of common laying ducks and their impacts on the phenotype of their mule ducklings. A total of 60 female laying ducks were divided into 2 dietary treatments at 10 wk of age. The restricted group received Met-restricted diets (R group) containing 0.25% of Met whereas the control group received control diets (C group) containing 0.40% of Met that meets Met requirements. The restriction was applied during the growing and laying periods, from 10 to 51 wk of age and a particular focus was put on female breeder traits that might be affected by the Met restriction. Plasma parameters of hepatic and lipid metabolisms were recorded in ducklings. Total weight (P < 0.001), albumen weight (P < 0.001) and albumen percentage of dry matter (P < 0.01) were decreased for eggs laid by female breeders from the R group. Both male and female ducklings from the R group of female breeders showed a reduced BW at hatching (P < 0.001) and a tendency to an increased proportional liver weight (P = 0.07). Finally, the maternal low dietary Met level modified plasma parameters in newborn ducklings regardless of sex: alkaline phosphatase (ALP) and alanine transaminase (ALT) activities were reduced (P = 0.07 and P = 0.002, respectively), levels of glucose (P = 0.03) and triglycerides (P = 0.01) were higher whereas level of free fatty acids decreased (P = 0.01). It was concluded that feeding female laying ducks with a restricted dietary Met content during the growing and laying periods has a negative effect on egg weight and composition. The ducklings that were restricted in nutrients during their early development, have a reduced BW, and altered lipid and hepatic metabolisms.

For many decades, prolificacy has been selected in meat sheep breeds as a polygenic trait but with limited genetic gain. However, the discovery of major genes affecting prolificacy has changed the way of selection for some ovine breeds implementing gene-assisted selection as in the French Lacaune and Grivette meat breeds, or in the Spanish Rasa Aragonesa breed. Based on statistical analysis of litter size parameters from 34 French meat sheep populations, we suspected the segregation of a mutation in a major gene affecting prolificacy in the Noire du Velay and in the Mouton Vendéen breeds exhibiting a very high variability of the litter size. After the genotyping of mutations known to be present in French sheep breeds, we discovered the segregation of the FecLL mutation at the B4GALNT2 locus and the FecXGr mutation at the BMP15 locus in Noire du Velay and Mouton Vendéen, respectively. The frequency of ewes carrying FecLL in the Noire du Velay population was estimated at 21.2% and the Mouton Vendéen ewes carrying FecXGr at 10.3%. The estimated mutated allele effect of FecLL and FecXGron litter size at +0.4 and +0.3 lamb per lambing in Noire du Velay and Mouton Vendéen, respectively. Due to the fairly high frequency and the rather strong effect of the FecLL and FecXGr prolific alleles, specific management programmes including genotyping should be implemented for a breeding objective of prolificacy adapted to each of these breeds.

The objective of this study was to evaluate genetic and non-genetic factors influencing artificial insemination (AI) success in French dairy goats. Data analysis, on a total of 584 676 and 386 517 AI records for Alpine and Saanen breed, respectively, collected from 1992 to 2009, was conducted separately on each breed. We used a linear simple repeatability animal model which combined male and female random effect and environmental fixed effects. The most important environmental factor identified was the period within year effect due to the European heat wave of 2003. The estimated values of the annual fertility exhibited a negative trend of 1% loss of AI success per 10 years for Alpine breed only. The range of variation for the flock×within years random effect was 70% and 65% for Alpine and Saanen breeds. The negative effect on AI success of antibody production after repetitive hormonal treatment was confirmed. We observed an important positive relationship between fertility and protein yield expressed as quartile within flock×years of protein 250-day yield for female with lactation number over 1, while this trend was negative for primiparous females. We detected a negative effect of the duration of conservation of semen with a difference of about 4% of AI success between extreme values (2 to 8+ or 9+ years). Heritability estimates for male fertility were 0.0037 and 0.0043 for Alpine and Saanen breed respectively, while estimates for female fertility was 0.040 and 0.049. Repeatability estimates for males were 0.008 and 0.010 for Alpine and Saanen, respectively, and 0.097 and 0.102 for females. With such low values of heritability, selection can hardly affect fertility.

Phenotype variability depends on genetics and environmental factors. Improving farm animal performances relies on genetic variability, but the possible improvement of selection schemes taking into account nongenetic transgenerational inheritance has become a topic of choice. Indeed, the parental diet may influence the adult phenotype of the offspring, and more and more studies suggest that information acquired from environmental exposures may be transmitted across generations. In this review, we focus on nongenetic inheritance of diet effects in birds, either as parental effect, that is, “nutritional programming,” or through the transmission of information across several generations, via “transgenerational epigenetic inheritance.” Compared to mammal models with regard to their closer proximity with humans, bird models have the added benefit to minimize maternal confounding effects by a direct manipulation of the egg content.

Polymorphisms in the gene encoding bone morphogenetic protein 15 (BMP15) have been associated with multiple ovulations in sheep. As BMP15 regulates inhibin expression in rodents, we assumed that the ovarian inhibin/activin system could mediate part of the effect of BMP15 mutations in the regulation of ovulation rate in sheep. To answer this question, we have studied the effects of two natural loss-of-function mutations of BMP15 on the expression of components of this system. The FecXR and the FecXGr mutations, when present respectively in Rasa Aragonesa ewes at the heterozygous state and in Grivette ewes at the homozygous state, were associated with a twofold increase in ovulation rate. There were only small differences between mutant and wild-type ewes for mRNA expression of INHA, INHBA, ACVR1B, ACVR2A, FST or TGFBR3 in granulosa cells and inhibin A or activin A concentrations in follicular fluid. Moreover, the effects of mutations differed between breeds. In cultures of granulosa cells from wild-type ewes, BMP15, acting alone or in synergy with GDF9, stimulated INHA, INHBA and FST expression, but inhibited the expression of TGFBR3. Activin A did not affect INHBA expression, but inhibited the expression of ACVR2A also. The complexity of the inhibin/activin system, including positive and antagonistic elements, and the differential regulation of these elements by BMP15 and activin can explain that the effects of BMP15 mutations differ when present in different genetic backgrounds. In conclusion, the ovarian inhibin/activin system is unlikely to participate in the increase of ovulation rate associated with BMP15 mutations in sheep.

International audience; AbstractBackground Classical genetic canalization models, which accommodate the mean and variance of a trait separately, provide a flexible approach to take heteroscedasticity for continuous traits into account. However, this model is not appropriate for discrete traits. The aim of this work was to propose heteroscedastic threshold models suitable for the genetic analysis of ordinal data.MethodsIn order to first fit the mean and variance of ordinal traits separately, we extended the classical threshold model (TM) for discrete data by introducing non-genetic and genetic factors of heterogeneity on the variance of its underlying variable, which leads to a homothetic threshold model HTM and its alternative parameterization HTM’ in which the thresholds of different individuals are linked by a homothetic-translation. Relaxing the constraint between the thresholds led us to propose an independent threshold model ITM that was more flexible than HTM’ but required the estimation of more parameters. TM, HTM and ITM models were applied to study 19,671 records on litter size in Romane sheep.ResultsBoth HTM and ITM were able to disentangle the link between the mean and variance that holds in the classical homoscedastic threshold model. The results obtained for the litter size of Romane ewes showed that the data was best fitted with HTM compared to ITM and TM. The correlations between the observed and predicted variances were equal to 0.6 and 0.2 for HTM and TM, respectively. These analyses showed the existence of a genetic component for the heterogeneity of litter size in sheep that was taken into account in HTM.ConclusionsHTM is the most suitable model to study the variability of litter size in sheep. It accommodates both the mean and variance separately while requiring the estimation of only a few parameters.

Major Genes Influencing Prolificacy in Sheep: A Source of Basic Knowledge of Ovarian Function and a Tool for Genetic Improvement. International Plant & Animal Genome XXIII conference

[EN] The aim of this preliminary experiment was to investigate the individual variability of the sexual receptivity of a rabbit doe in the presence of a buck (lordosis position, 0-1 variable). Twenty primiparous does maintained without reproduction were tested over 4 mo (3 tests per week at a 2- or 3-day interval) after their first kindling. Out of 48 tests, the receptivity rate was 52.5±50.0% on average and varied from 20.0 to 73.3% depending on the test day. The does were lactating at the beginning of the test period and a strong receptivity decrease was revealed at the peak of lactation. Receptivity did not vary according to the tester buck or to the test operator. The individual receptivity of does varied from 8.6 to 81.3%; three of them had a receptivity rate lower than 30% and four of them a receptivity rate greater than 70%. No relationship was revealed between average receptivity and body weight or body weight variations around first litter weaning. The repeatability of sexual receptivity of non-lactating does was 23.2%. Lowly receptive does had a shorter average oestrus time (4 tests) and a shorter dioestrus time (≤3 tests). The correlation between average receptivity and average oestrus time was 0.80. These results indicate a fairly high individual variability of the expression of rabbit sexual receptivity and of its duration, and justify the exploration of an eventual genetic origin in a subsequent experiment., The authors are grateful to P. Aymard, J.M. Bonnemère, Annette Debrusse, H. Girotto and D. Labatut, technicians at the PECTOUL Experimental Rabbit Farm at INRA in Toulouse, and to J. Ruesche and the SAGA IT team for their collaboration, and G. Wagman for her careful revision of the English.

Background In mammals, multigenerational environmental effects have been documented by either epidemiological studies in human or animal experiments in rodents. Whether such phenomena also occur in birds for more than one generation is still an open question. The objective of this study was to investigate if a methionine deficiency experienced by a mother (G0) could affect her grand-offspring phenotypes (G2 hybrid mule ducks and G2 purebred Muscovy ducks), through their Muscovy sons (G1). Muscovy drakes are used for the production of mule ducks, which are sterile offspring of female common duck (Anas platyrhynchos) and Muscovy drakes (Cairina moschata). In France, mule ducks are bred mainly for the production of “foie gras”, which stems from hepatic steatosis under two weeks of force-feeding (FF). Two groups of female Muscovy ducks received either a methionine deficient diet or a control diet. Their sons were mated to Muscovy or to common duck females to produce Muscovy or Mule ducks, respectively. Several traits were measured in the G2 progenies, concerning growth, feed efficiency during FF, body composition after FF, and quality of foie gras and magret. Results In the G2 mule duck progeny, grand-maternal methionine deficiency (GMMD) decreased 4, 8, and 12 week body weights but increased weight gain and feed efficiency during FF, and abdominal fat weight. The plasmatic glucose and triglyceride contents at the end of FF were higher in the methionine deficient group. In the G2 purebred Muscovy progeny, GMMD tended to decrease 4 week body weight in both sexes, and decreased weight gain between the ages of 4 and 12 weeks, 12 week body weight, and body weight at the end of FF in male offspring only. GMMD tended to increase liver weight and increased the carcass proportion of liver in both sexes. Conclusion Altogether, these results show that the mother’s diet is able to affect traits linked to growth and to lipid metabolism in the offspring of her sons, in Muscovy ducks. Whether this transmission through the father of information induced in the grand-mother by the environment is epigenetic remains to be demonstrated. Electronic supplementary material The online version of this article (doi:10.1186/s12863-015-0303-z) contains supplementary material, which is available to authorized users.

[EN] Sexual receptivity of rabbit does at insemination greatly influences fertility and is generally induced by hormones or techniques known as “biostimulation”. Searching for more sustainable farming systems, an original alternative would be to utilise the genetic pathway to increase the does’receptivity. The purpose of the present study was to identify genetic and non-genetic factors that influence rabbit doe sexual receptivity, in the context of a divergent selection experiment over 1 generation. The experiment spanned 2 generations: the founder generation (G0) consisting of 140 rabbit does, and the G1 generation comprising 2 divergently selected lines (L and H lines) with 70 does each and 2 successive batches from each generation. The selection rate of the G0 females to form the G1 lines was 24/140. The selection tests consisted of 16 to 18 successive receptivity tests at the rate of 3 tests per week. On the basis of 4716 tests from 275 females, the average receptivity was 56.6±48.2%. A batch effect and a test operator effect were revealed. The contribution of females to the total variance was 20.0%, whereas that of bucks was only 1.1%. Throughout the experiment, 18.2% of does expressed a low receptivity (< 34%), 50.7% a medium one and 33.1% a high one (>66%). Some does were frequently receptive, whereas others were rarely receptive. The repeatability of sexual receptivity was approximately 20%. The results confirmed the high variability of sexual receptivity of non-lactating rabbit does maintained without any biostimulation or hormonal treatment. A lack of selection response on receptivity was observed. Accordingly, the heritability of receptivity was estimated at 0.01±0.02 from an animal model and at 0.02±0.03 from a sire and dam model. The heritability of the average receptivity of a doe was calculated as 0.04. In agreement with the low estimated heritability, the heritability determined was no different from zero. Nevertheless, the occurrence of pseudopregnancies due to uncontrolled ovulations and the presence of corpora lutea, as assessed by progesterone titrations, could have interfered with receptivity. Further studies would be necessary to confirm the low heritability of female rabbit receptivity., The authors are grateful to the Department of Animal Genetics of the National Institute of Agronomical Research for its financial contribution, to P. Aymard, J.M. Bonnemère, Annette Debrusse, H. Girotto and D. Labatut, technicians at the PECTOUL Experimental Rabbit Farm at INRA in Toulouse, to J. Ruesche, the IT team from SAGA for their collaboration and to G. Wagman for her careful linguistic revision.

Inheritance of the ovulation rate (OR) in the Lacaune meat breed was studied through records from a small nucleus of 36 hyper-prolific ewes screened on farms on the basis of their natural litter size, and from progeny data of three selected Lacaune sires. These sires were chosen at the AI centre according to their breeding values estimated for the mean and the variability of their daughters' litter size. Non-carrier Lacaune dairy ewes were inseminated to produce 121 F1 daughters and 27 F1 sons. Twelve sons (four from each sire) were used in turn to inseminate non-carrier Lacaune dairy ewes providing 260 BC progeny ewes. F1 and BC progeny were brought from private farms and gathered after weaning on an experimental farm where ovulation rates were recorded in the first and second breeding seasons. With an average of 6.5 records each, the mean OR of hyper-prolific ewes was very high (5.34), and 38.4% of records showed a rate of 6 or more. F1 data showed high repeatability of OR (r = 0.54) within ewe, with significant variability among ewes. High OR (≥ 4) were observed in each family. A segregation analysis provided a significant likelihood ratio and classified the three founders as heterozygous. BC ewes also displayed high repeatability of OR (r = 0.47) and the mean OR varied considerably between families (from 1.24 to 1.78). Seven of the 12 BC families presented high-ovulating ewes (at least one record ≥ 4) and segregation analysis yielded a highly significant likelihood ratio as compared to an empirical test distribution. The high variability of the mean ovulation rate shown by a small group of daughters of BC ewes inseminated by putative carrier F1 rams, and the very high ovulation rate observed for some of these ewe lambs, confirmed the segregation of a major gene with two co-dominant alleles borne by an autosome. The difference between homozygous non-carriers and heterozygous ewes was about one ovulation on the observed scale and 2.2 standard deviations on the underlying scale.

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Chantier qualité GA; International audience; The "ram effect" has been an alternative method among many techniques to control the period of sexual activity in sheep. The objectives of this study were to (1) study the reproductive performances of the Barbarine breed in the spring, (2) to evaluate females' response to the ram effect, and estimate the phenotypic and genetic parameters of the females' response to the "ram effect". A total of 16,150 matings of 4201 ewes recorded during a period of ten years in seven flocks was used in this study. Among these ewes 1335 have known parents and were born from 199 sires and 1053 dams. Four cyclicity traits were analyzed: spontaneous, normal, short and not answer to the ram effect. The main results show that approximately 30% of the females were cyclic out of season and more than 60% became cyclic with the "ram effect". Flock, year and age at lambing had significant effects on the traits analyzed. A high negative phenotypic correlation of -0.75 was found between responses of normal cycles and responses of short cycle. The spontaneous ovarian activity in the spring was negatively correlated with the absence of response to a ram effect (r(g) = -0.44) and positively correlated to the response by a normal cycle (r(g) = 0.63). The correlation between responses with the normal cycle and short cycle was highly negative (r(g) = -0.88). For all variables, heritability was low (from 0.03 to 0.09) but significantly different from 0 and repeatability was slightly higher and varied from 0.06 to 0.1. These results indicate the great opportunity offered by the Barbarine breed for out of season breeding. (C) 2013 Elsevier B.V. All rights reserved.

Prolific sheep have proven to be a valuable model to identify genes and mutations implicated in female fertility. In the Lacaune sheep breed, large variation in litter size is genetically determined by the segregation of a fecundity major gene influencing ovulation rate, named FecL and its prolific allele FecLL. Our previous work localized FecL on sheep chromosome 11 within a locus of 1.1 Mb encompassing 20 genes. With the aim to identify the FecL gene, we developed a high throughput sequencing strategy of long-range PCR fragments spanning the locus of FecLL carrier and non-carrier ewes. Resulting informative markers defined a new 194.6 kb minimal interval. The reduced FecL locus contained only two genes, insulin-like growth factor 2 mRNA binding protein 1 (IGF2BP1) and beta-1,4-N-acetyl-galactosaminyl transferase 2 (B4GALNT2), and we identified two SNP in complete linkage disequilibrium with FecLL. B4GALNT2 appeared as the best positional and expressional candidate for FecL, since it showed an ectopic expression in the ovarian follicles of FecLL/FecLL ewes at mRNA and protein levels. In FecLL carrier ewes only, B4GALNT2 transferase activity was localized in granulosa cells and specifically glycosylated proteins were detected in granulosa cell extracts and follicular fluids. The identification of these glycoproteins by mass spectrometry revealed at least 10 proteins, including inhibin alpha and betaA subunits, as potential targets of B4GALNT2 activity. Specific ovarian protein glycosylation by B4GALNT2 is proposed as a new mechanism of ovulation rate regulation in sheep, and could contribute to open new fields of investigation to understand female infertility pathogenesis., Author Summary Prolific sheep have proven to be a valuable model to identify genes and mutations implicated in ovarian function and female fertility. Indeed, fecundity genes of the Bone Morphogenetic Protein (BMP) family discovered in sheep was evidenced as genetic candidates to explain female infertility pathologies. Studying French Lacaune sheep breed, we discovered another fecundity gene named B4GALNT2, encoding a glycosylation enzyme that is not related to the BMP family. The high prolificacy of Lacaune sheep was explained by overexpression of B4GALNT2 in the ovary leading to atypical glycosylation of inhibin, an important hormone regulating ovarian function. Our findings open promising fields of investigation to better understand female fertility disorders.

International audience

Background Successful achievement of early folliculogenesis is crucial for female reproductive function. The process is finely regulated by cell-cell interactions and by the coordinated expression of genes in both the oocyte and in granulosa cells. Despite many studies, little is known about the cell-specific gene expression driving early folliculogenesis. The very small size of these follicles and the mixture of types of follicles within the developing ovary make the experimental study of isolated follicular components very difficult. The recently developed laser capture microdissection (LCM) technique coupled with microarray experiments is a promising way to address the molecular profile of pure cell populations. However, one main challenge was to preserve the RNA quality during the isolation of single cells or groups of cells and also to obtain sufficient amounts of RNA. Using a new LCM method, we describe here the separate expression profiles of oocytes and follicular cells during the first stages of sheep folliculogenesis. Results We developed a new tissue fixation protocol ensuring efficient single cell capture and RNA integrity during the microdissection procedure. Enrichment in specific cell types was controlled by qRT-PCR analysis of known genes: six oocyte-specific genes (SOHLH2, MAEL, MATER, VASA, GDF9, BMP15) and three granulosa cell-specific genes (KL, GATA4, AMH). A global gene expression profile for each follicular compartment during early developmental stages was identified here for the first time, using a bovine Affymetrix chip. Most notably, the granulosa cell dataset is unique to date. The comparison of oocyte vs. follicular cell transcriptomes revealed 1050 transcripts specific to the granulosa cell and 759 specific to the oocyte. Functional analyses allowed the characterization of the three main cellular events involved in early folliculogenesis and confirmed the relevance and potential of LCM-derived RNA. Conclusions The ovary is a complex mixture of different cell types. Distinct cell populations need therefore to be analyzed for a better understanding of their potential interactions. LCM and microarray analysis allowed us to identify novel gene expression patterns in follicular cells at different stages and in oocyte populations.

Two models can be used for studying binary results of AI. The additive threshold model proposes an underlying variable as summing the environmental and genetic effects from the 2 individuals involved in the mating, and the product threshold model assumes that the conditional probability of AI success is the product of the probabilities of success of 2 unobserved binary phenotypes (one is the male fertility; the other is the female fertility). The purpose of this paper is to compare the predictive ability of the product and the additive threshold models for studying AI results and to compare results obtained with the 2 models in 3 different species cattle, sheep, and rabbits. Results showed that the predictive ability of the product model is similar to the additive model in sheep and rabbits but worst in cattle (percentage of wrong prediction = 42, 27, and 35% in the additive model; 43, 28, and 47% in the product model in sheep, rabbits, and cattle, respectively). Even when the 2 models have similar performance, they differed in their EBV (for instance, Pearson correlation between EBV predicted with the 2 models = 0.46 in sheep for male fertility). The product model can determine which sex is responsible for an AI failure. In sheep, the female was the responsible in 94% of the cases and male in 2% of them; in rabbits, the female was the responsible in 54% of the cases and the male in 39% of them. Different estimates of probabilities for male and female fertility success obtained with the product model in the 3 species suggest that male and female fertilities behave differently depending on the species and the uniqueness of the data sets. Although product model seems to provide additional information in the fertility process, further research is needed to understand the worst performance of the product model in cattle. © 2011 American Society of Animal Science. All rights reserved.

Spontaneous ovulatory activity (SOA) in spring has been used to study the out-of-season breeding ability of Merinos d’Arles (MA) ewes. Within this breed, an association was found between more intense seasonality and genotype 2/2 at a MnlI restriction site (allele 2 for its absence v. 1 for its presence) in Exon II of the MT1 receptor gene. This study was designed to ascertain whether this association results in a direct effect of the MT1 genotype on the expression of seasonality in MA ewes. In the first year of the study, genotyping of 314 MA ewes at locus MnlI was carried out and resulted in frequencies of 43.0%, 44.9% and 12.1% for genotypes 1/1, 1/2 and 2/2, respectively. The SOA of these ewes was determined in early April of two consecutive years by assaying plasma progesterone concentrations in two blood samples taken 9 days apart. Groups of 30 ewes of each homozygous genotype (1/1 and 2/2) were identified from this population and their SOA was followed by taking blood samples at regular intervals between January and mid-April of the second and third year of the study. In the second year, groups of ewes were managed together on rangelands, whereas in the third year each group was split into two subgroups given differential feed levels. The results clearly showed that genotype had no significant effect on SOA during the 2- to 3-month period preceding the introduction of rams for spring mating. In the second year of the study, in which the experimental procedure allowed a fair comparison of the fertility of ewes in spring mating, fertility was similar for both genotypes. The reciprocity of the association was not demonstrated and the MnlI polymorphic site could not be used as a genetic marker of selection for out-of-season breeding ability, at least not in the MA breed. The percentage of cycling ewes significantly decreased between January and April, and older ewes (5 or 6 years old depending on the year of the study) were more cyclic than younger ones (2 and 3 years old, respectively). The differential feeding level of ewes from early February did not significantly affect their SOA during the time period studied in the third year of the study.

SUMMARY In the past, several studies have been performed to look for genes affecting traits of interest using microsatellite markers in sheep. Even if numerous chromosomal regions have been detected, few genes and causal mutations have been identified. The recent availability of high density SNP chip in sheep is a technological revolution for genomic researches and breeding selection. Firstly, genomic research in small ruminants will be boosted towards the identification of causative mutations underlying large genetic effects on sustainability traits, or the identification of very closely linked genetic markers that allow selecting genes in ovine breeding programmes. Secondly, such new molecular tools allow considering a new type of selection: the genomic selection. The principle of this selection is to use marker effects (without any information about the underlying genes) estimated within a part of the population which is phenotyped and genotyped (called the training population) and applying these effects to the rest of the genotyped population. However, the genomic selection might not always be a profitable strategy in sheep.

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International audience; In the Lacaune sheep population, two major loci influencing ovulation rate are segregating: FecX and FecL. The FecXL mutation is a non-conservative substitution (p.Cys53Tyr) in BMP15 that prevents the processing of the protein. Using a statistical approach, FecL has been shown to be an autosomal major gene. A full genome scan localized the FecL locus on sheep chromosome 11. Fine mapping reduced the interval containing FecL to markers BM17132 and FAM117A, corresponding to a synteny block of 1.1 megabases on human chromosome 17, which encompasses 20 genes. The expression of 16 genes from this interval was observed in tissues of the reproductive axis, but expression was not affected in homozygous FecLL females. In this interval, a unique haplotype was associated with the FecLL mutation. This particular haplotype could be predicted by the DLX3:c.*803A > G SNP in the 3' UTR sequence of the DLX3 gene. This SNP provided accurate classification of animals (99.5%) as carriers or non-carriers of the mutation and therefore maybe useful in marker assisted selection. A synergistic action of FecLL and FecXL mutations on both ovulation rate and litter size was demonstrated. Until now, all the Fec genes identified in sheep belong to the bone morphogenetic protein (BMP) system. Based on the human orthologous region, none of the 20 genes in the FecL region corresponds to known molecules in the BMP system. The identification of the FecLL mutation could lead to the discovery of a new pathway involved in the regulation of ovulation rate.

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Artificial inseminations (n = 678 168) recorded during 5 years in five French artificial insemination (AI) centres (2 'Lacaune', 1 'Manech tête rousse', 1 'Manech tête noire' and 1 'Basco béarnaise') were analysed to determine environmental and genetic factors affecting the insemination results. Analyses within centre-breed were performed using a linear model, which jointly estimates male and female fertility. This model combined four categories of data: the environmental effects related to the female, those related to the male, the non-sex-specific effects and finally the pedigree data of these males and females. After selection, the environmental female effects considered were age, synchronisation (0/1) on the previous year, total number of synchronisations during the female reproductive life, time interval between previous lambing and insemination, already dry or still lactating (0/1) when inseminated, and milk quantity produced during the previous year expressed as quartiles intra herd * year. The environmental male effects were motility and concentration of the semen. The non-sex-specific effects were the inseminator, the interaction herd * year nested within the inseminator, considered as random effects and the interaction year * season considered as a fixed effect. The main variation factors of AI success were relative to non-sex-specific effects and to female effects. Heritability estimates varied from 0.001 to 0.005 for male fertility and from 0.040 to 0.078 for female fertility. Repeatability estimates varied from 0.007 to 0.015 for male fertility and from 0.104 to 0.136 for female fertility. These parameters indicate that genetic improvement of AI results through a classical polygenic selection would be difficult. Moreover, in spite of the large quantity of variation factors fitted by the joint model, a very large residual variance remained unexplained.

A total of 416,670 lactations for 189,101 ewes from 3,603 sires and distributed across 1,978 herd-year groups were used to estimate genetic and environmental parameters of standardized milk yield (SMY(T)), fertility in ewe lambs (PR(1)), and fertility in adult ewes (PR(A)). Parameters were estimated with a multiple-trait sire linear model. Heritabilities for SMY(T), PR(1), and PR(A) were 0.27 (0.009), 0.04 (0.004), and 0.05 (0.004), respectively. These results were in accordance with the literature. The genetic correlation between PR(1) and PR(A) was 0.55, indicating that fertility is not the same trait in ewe lambs and adult ewes. The genetic correlation between milk yield and lamb fertility was not significantly different from zero. The genetic correlation between milk yield and fertility in adult ewe (-0.23) was in the range of antagonistic correlations reported in dairy cattle. Consequently, these results show that selection for milk yield can induce an indirect decrease in fertility. Nevertheless, no phenotypic decrease in fertility in artificial insemination matings has been observed in this population. This is the first time that correlation between milk yield and fertility is reported in sheep and further investigations are needed to confirm this result.

A divergent selection experiment based on the homogeneity of birth weight in rabbit was carried out at the INRA experimental farm of Auzeville. The two lines were created by selecting breeding does and bucks from the female strain AGP22 bred at the Grimaud Freres Selection company for environmental variability of birth weight. This involved a new model incorporating a genotypic value for the mean and a genotypic value for the environmental variance. There was a selection response with a significant difference in the within-litter standard deviation of birth weight between the lines after four generations of selection (6.93 g vs. 8.13 g). The mortality of kits at birth and from birth to weaning was lower in the “homogeneous” line than in the “heterogeneous” line (16.6% vs. 18.6% and 17.7% vs. 32.7%, respectively). The litter size at weaning was higher in the “homogeneous” line (7.22 vs. 5.39). The selection on the homogeneity of birth weight had no significant influence on the other traits. It was concluded that selection for reduced environmental variance of birth weight improved young rabbit survival without reducing the mean of birth weight.

Genetic mutations with major effects on ovulation rate and litter size in sheep were recently identified in three genes belonging to the TGFβ superfamily pathway: the bone morphogenetic protein 15 (BMP15, also known as GDF9b), growth differentiation factor 9 (GDF9), and BMP receptor type IB (also known as activin-like kinase 6). Homozygous BMP15 or GDF9 mutations raise female sterility due to a failure of normal ovarian follicle development, whereas heterozygous animals for BMP15 or GDF9 as well as heterozygous and homozygous animals for BMP receptor type IB show increased ovulation rates. In the present work, a new naturally occurring mutation in the BMP15 gene in the high prolific Lacaune sheep breed is described. The identified variant is a C53Y missense nonconservative substitution leading to the aminoacidic change of a cysteine with a tyrosine in the mature peptide of the protein. As for other mutations found in the same gene, this is associated with an increased ovulation rate and sterility in heterozygous and homozygous animals, respectively. Further in vitro studies showed that the C53Y mutation was responsible for the impairment of the maturation process of the BMP15 protein, resulting in a defective secretion of both the precursor and mature peptide. Overall, our findings confirm the essential role of the BMP15 factor in the ovarian folliculogenesis and control of ovulation rate in sheep.

Data from 51 107 and 11 839 ejaculates collected on rams of the "Lacaune" and "Manech tete rousse" breeds, respectively, were analysed to determine environmental and genetic factors affecting semen production traits (ejaculate volume, semen concentration, number of spermatozoa and motility) in young (≤1 year) and adult (≥2 years) rams. Fixed effects and variance components were estimated using multiple trait animal models within each breed. For all traits, the main environmental effects identified were year, season, number of ejaculations, daily variation, interval from previous to current collection and age. Heritability estimates were moderate for volume, concentration and number of spermatozoa (0.12 to 0.33) and lower for motility (0.02 to 0.14). Genetic correlations between ages differed from 1 for all traits (0.14 to 0.90), indicating that semen characteristics corresponded to different traits in young and adult rams. Genetic and phenotypic correlations among traits within age category were globally similar for the different breeds and categories of animals.

The litter size in Suffolk and Texel-sheep was analysed using REML and Bayesian methods. Litters born after hormonal induced oestrus and after natural oestrus were treated as different traits in order to estimate the genetic correlation between the traits. Explanatory variables were the age of the ewe at lambing, period of lambing, a year*flock-effect, a permanent environmental effect associated with the ewe, and the additive genetic effect. The heritability estimates for litter size ranged from 0.06 to 0.13 using REML in bi-variate linear models. Transformation of the estimates to the underlying scale resulted in heritability estimates from 0.12 to 0.17. Posterior means of the heritability of litter size in the Bayesian approach with bi-variate threshold models varied from 0.05 to 0.18. REML estimates of the genetic correlations between the two types of litter size ranged from 0.57 to 0.64 in the Suffolk and from 0.75 to 0.81 in the Texel. The posterior means of the genetic correlation (Bayesian analysis) were 0.40 and 0.44 for the Suffolk and 0.56 and 0.75 for the Texel in the sire and animal model respectively. A bivariate threshold model seems appropriate for the genetic evaluation of prolificacy in the breeds concerned. ispartof: Genetics, selection, evolution. : GSE vol:36 issue:5 pages:543-62 ispartof: location:France status: published

It has been demonstrated that variations in litter size or ovulation rate in different breeds of sheep can be associated with the segregation of several major genes. This set of natural mutants constitutes a valuable resource to determine key points in the biochemical pathways controlling the development of ovarian follicles. The French genetic programmes were devised to identify two of these genes: the Booroola (FecB) and Lacaune genes. The FecB prolific mutation corresponds to a non-conservative mutation (Q249R) in the intracellular kinase-signalling domain of the bone morphogenetic protein receptor type IB (BMPR-IB) gene. The Lacaune gene is situated on ovine chromosome 11. Positional cloning is currently in progress to identify the relevant gene and mutation. A similar approach, limited to linkage testing of candidate genes, is proposed to classify the different prolificacy genes in sheep.

Ewes from the Booroola strain of Australian Mérino sheep are characterized by high ovulation rate and litter size. This phenotype is due to the action of the FecB B allele of a major gene named FecB , as determined by statistical analysis of phenotypic data. By genetic analysis of 31 informative half-sib families from heterozygous sires, we showed that the FecB locus is situated in the region of ovine chromosome 6 corresponding to the human chromosome 4q22–23 that contains the bone morphogenetic protein receptor IB ( BMPR-IB ) gene encoding a member of the transforming growth factor-β (TGF-β) receptor family. A nonconservative substitution (Q249R) in the BMPR-IB coding sequence was found to be associated fully with the hyperprolificacy phenotype of Booroola ewes. In vitro , ovarian granulosa cells from FecB B / FecB B ewes were less responsive than granulosa cells from FecB + / FecB + ewes to the inhibitory effect on steroidogenesis of GDF-5 and BMP-4, natural ligands of BMPR-IB. It is suggested that in FecB B / FecB B ewes, BMPR-IB would be inactivated partially, leading to an advanced differentiation of granulosa cells and an advanced maturation of ovulatory follicles.

International audience; Classical selection for increasing prolificacy in sheep leads to a concomitant increase in its variability, even though the objective of the breeder is to maximise the frequency of an intermediate litter size rather than the frequency of high litter sizes. For instance, in the Lacaune sheep breed raised in semi-intensive conditions, ewes lambing twins represent the economic optimum. Data for this breed, obtained from the national recording scheme, were analysed. Variance components were estimated in an infinitesimal model involving genes controlling the mean level as well as its environmental variability. Large heritability was found for the mean prolificacy, but a high potential for increasing the percentage of twins at lambing while reducing the environmental variability of prolificacy is also suspected. Quantification of the response to such a canalising selection was achieved.

To determine whether a link exists between reproductive seasonality and the structure of the gene for melatonin receptor Mel(1a), the latter was studied in two groups of Mérinos d'Arles (MA) ewes previously chosen for their genetic value, which took into account their own out-of-season ovulatory activity adjusted by environmental parameters and that of their relatives. The genomic DNA of 36 ewes found regularly cycling in spring (group H) and that of 35 ewes never cycling in spring (group L) during the 2-3 yr before the present study was prepared, and the cDNA corresponding to almost all exon II was amplified and checked for the presence of MnlI restriction sites. The presence (+) or absence (-) of an MnlI site at position 605 led to genotypes "++", "+-", and "--", whose frequencies differed significantly (P0.001) between the H and L groups: 52.8%, 47.2%, and 0% vs. 28.5%, 42.9%, and 28.5%, respectively. Sequencing of exon II cDNA in group L ewes with genotype -- showed the presence of only one allele - with 4 mutations, while that in ewes with genotype ++ showed different types of alleles unrelated to the H or L groups. These + alleles exhibited a combination of 1 to 7 of the 8 mutations recorded in the part of exon II studied. The genotyping of 29 ewes from the more seasonal Ile-de-France breed indicated that 38% of animals had a -- genotype and exhibited the same mutations as in the MA ewes. Finally, a comparison of (125)I-melatonin binding to membrane preparations of pars tuberalis showed a lower number of binding sites (P0. 0005) in MA ewes with genotype ++ than in those with genotype -- (43. 2 +/- 4.4 vs. 75.4 +/- 8.4 fmol/mg protein in genotype ++ and genotype --, respectively). In conclusion, the data show an association between genotype -- for site MnlI at position 605 and seasonal anovulatory activity in MA ewes.

The genetic parameters of spontaneous spring ovulatory activity were investigated in the Mérinos d’Arles breed under the usual pastoral and transhumant management conditions of this breed in southeastern France. Ovulatory activity was determined by assaying the plasma progesterone concentration in two blood samples taken 8-10 days apart. The data set consisted of 1 887 ovulatory activity performance measurements in 1995, 1996 and 1997 on 933 ewes, daughters of 176 rams. The effects of the ’physiological status’ (hoggets, adult ewes with or without lambing in the previous autumn), age and live weight just before the mating period were found to be highly significant. They were included in the linear animal model and the threshold sire model used to estimate genetic parameters. On average, 27.9 % of ewes exhibited ovulatory activity in April. Age and live weight just before the mating period had a marked positive effect on ovulatory activity. A difference of about 8-9 % was observed between extreme classes for these factors. The heritability and repeatability estimated through the linear model were 0.20 (standard error: 0.04) and 0.30 (0.07), respectively. When using the threshold model, the heritability was 0.37. These values led us to conclude that a genetic approach for improving spontaneous spring ovulatory activity should be further developed. Nevertheless, further studies are necessary to determine all the implications of such selection., Les paramètres génétiques de l’activité ovulatoire spontanée au printemps ont été estimés en race Mérinos d’Arles dans le système d’élevage pastoral traditionnel (transhumance estivale) du sud-est de la France. Le dosage de la progestérone plasmatique dans deux prélèvements sanguins effectués à 8-10 j d’intervalle a permis de déterminer l’activité ovulatoire des brebis. 1887 performances d’activité ovulatoire ont été enregistrées en 1995, 1996 et 1997, sur 933 brebis issues de 176 béliers. Le «statut physiologique » (antenaises, brebis adultes avec ou sans mise bas à l’automne précédent), l’âge et le poids au moment de la lutte des brebis ont des effets très significatifs sur l’activité ovulatoire. Ils ont été pris en compte dans le modèle animal linéaire et le modèle père à seuil utilisés pour estimer les paramètres génétiques. En moyenne, 27,9 % des brebis présentaient une activité ovulatoire en avril. L’âge et le poids au moment de la lutte ont un net effet positif sur l’activité ovulatoire. Une différence de 8-9 % a été observée entre les classes extrêmes pour ces facteurs. L’héritabilité et la répétabilité estimées avec le modèle linéaire sont de 0,20 (erreur standard : 0,04) et de 0,30 (0,07), respectivement. L’héritabilité calculée avec le modèle à seuil est de 0,37. En conclusion, compte tenu de ces valeurs, l’approche génétique visant à améliorer l’activité ovulatoire spontanée au printemps mérite d’être poursuivie. Néanmoins, d’autres études sont nécessaires pour connaître toutes les implications que supposent une telle sélection.

*INRA Station d'Amélioration Génétique des Animaux 31326 Castanet-Tolosan (FRA) Diffusion du document : INRA Station d'Amélioration Génétique des Animaux 31326 Castanet-Tolosan (FRA); International audience

The present study was conducted to assess the degree of genetic determination of the variability in the mean nocturnal plasma concentration of melatonin in sheep. Three hundred twelve ewes born from 18 males and with known genealogy were sampled at the summer and the winter solstices. The nocturnal plasma melatonin concentration was defined as the mean of four plasma samples taken at hourly intervals in the middle of the night (2200–0200). Identity of the father ( P < 0.001) and the solstice ( P < 0.05) were significant. Melatonin concentrations varied considerably among individuals [338.4 ± 197.5 (SD) pg/ml; range 26.6–981.3 pg/ml] and between rams regarding the melatonin concentrations of their daughters (range from 202.9 to 456.3 pg/ml). Inheritance was analyzed by a statistical model that allows discrimination of genetic effects from nongenetic effects and that estimates repeatability and heritability coefficients. Both the repeatability coefficient between solstices (0.60) and heritability coefficient [0.45 ± 0.07 (SE)] were high. These results demonstrate that the variability in plasma melatonin concentration in ewes is under strong genetic control.

We investigated whether the genetic difference in plasma melatonin concentration in ewes was due to differences in the synthesis pathway from the pineal gland or in the catabolism of the hormone. Two groups of ewes [9 low (L) and 10 high (H)] were selected according to the breeding value of their mean nighttime plasma melatonin concentrations estimated at winter and summer solstices. In response to an identical dose of melatonin administered intravenously at 9:00 AM, no differences between groups were observed for any of the kinetic parameters: clearance rate, steady-state volume of distribution, terminal half-lives, and mean residence times. In the second experiment, two series of frequent blood samples were performed, one in the middle of the dark phase with samples taken every 5 min, and the other over 24 h with hourly samples. Highly significant differences between groups in nocturnal melatonin production rate were observed (L: 25.7 ± 2.8 vs. H: 63.1 ± 8.9 μg ⋅ kg−1⋅ h−1, P < 0.01). Thus the genetic differences in plasma melatonin concentrations in ewes originate in the synthesis pathway of the melatonin from the pineal gland rather than from differences in the catabolism of the hormone.

The ovulatory response of ewes from breeds that differ widely in prolificacy (Ile-de-France, ++ Booroola Merino, Romanov, F+ Booroola Merino and F+ Booroola Romanov with adult ovulation rates of about 1.5, 1.2, 3, 3 and 3.5 respectively) to 750 IU of hCG given at different physiological stages (before puberty, during anestrus or during the luteal phase) was compared. In all except one experiment, hCG induced ovulation in 73 to 98% of the lambs, indicating that follicles sensitive to LH were present at all stages studied. Ranking of the breeds according to hCG-induced ovulation rate in prepuberal lambs was similar to that based on adult ovulation rate. Furthermore, hCG induced more ovulations in prepuberal F+ than in ++ lambs (3.7 +/- 1.4 vs 1.7 +/- .8 at 4.5 mo of age) as well as in anestrous ewes (F + at 3.1 +/- 1.8 vs ++ at 1.6 +/- .7). Within ewes, the correlation between hCG-induced ovulation rate and mature ovulation rate was positive in nonprolific breeds but not significant in prolific breeds. We conclude that 1) the number of hCG-induced ovulations can be used to identify sheep that are carriers of the Booroola gene and 2) the mechanisms responsible for a number of large ovulatory follicles typical of a breed are present at stages (prepuberal, anestrus, luteal phase) other than the follicular phase.

Data from 51 107 and 11 839 ejaculates collected on rams of the "Lacaune" and "Manech tête rousse" breeds, respectively, were analysed to determine environmental and genetic factors affecting semen production traits (ejaculate volume, semen concentration, number of spermatozoa and motility) in young (≤1 year) and adult (≥2 years) rams. Fixed effects and variance components were estimated using multiple trait animal models within each breed. For all traits, the main environmental effects identified were year, season, number of ejaculations, daily variation, interval from previous to current collection and age. Heritability estimates were moderate for volume, concentration and number of spermatozoa (0.12 to 0.33) and lower for motility (0.02 to 0.14). Genetic correlations between ages differed from 1 for all traits (0.14 to 0.90), indicating that semen characteristics corresponded to different traits in young and adult rams. Genetic and phenotypic correlations among traits within age category were globally similar for the different breeds and categories of animals.

Canalising selection is handled by a heteroscedastic model involving a genotypic value for the mean and a genotypic value for the log variance, associated with a single phenotypic value. A selection objective is proposed as the expected squared deviation of the phenotype from the optimum, of a progeny of any candidate for selection. Indices and approximate expressions of parent-offspring regression are derived. Simulations are performed to check the accuracy of the analytical approximation. Examples of fat to protein ratio in goat milk yield and muscle pH data in pig breeding are provided in order to investigate the ability of these populations to be canalised towards an economic optimum, Le problème de la sélection canalisante est traité grâce à un modèle hétéroscédastique mettant en jeu une valeur génétique pour la moyenne et une valeur génétique pour le logarithme de la variance, toutes deux associées à une seule valeur phénotypique. Pour un objectif de sélection visant à minimiser l’espérance des carrés des différences entre le phénotype et l’optimum, pour un descendant d’un candidat à la sélection, des index sont estimés et des expressions approchées de la régression parent-descendant sont calculées. La précision de ces expressions analytiques est mesurée à l’aide de simulations. Afin d’appréhender la capacité de ces populations à être canalisées vers un optimum économique, des exemples sont donnés : le rapport entre matière grasse et matière protéique du lait de chèvre, et le pH d’un muscle chez le porc