Regulation of maternal behavior, social isolation responses, and postpartum estrus by steroid hormones and vaginocervical stimulation in sheep.

Periparturient ewes display several activities in addition to maternal behavior per se. They isolate themselves from the flock and, contrary to other mammals, do not show postpartum estrus. We investigated the possibility of a common hormonal control of maternal behavior, prepartum social isolation responses, and absence of postpartum estrus. We quantified responses to flockmate separation and incidence of sexual receptivity at several reproductive stages in intact ewes (Experiment 1). Responses to social isolation were lowest in preparturient ewes and at pregnancy day 149, intermediate at pregnancy day 147 and highest at day 136 and in non-pregnant ewes (P < 0.05 between the 3 levels). In a second experiment, we quantified the same parameters and maternal behavior in 1) ovariectomized ewes receiving medroxyprogesterone acetate only (ovxMPA); 2) ovariectomized ewes receiving MPA + estradiol benzoate (ovxSHORT); 3) intact ewes receiving a longer MPA + estradiol dipropionate treatment, before and after vaginocervical stimulation (VCS). Before VCS no steroid treatments decreased social isolation responses and maternal behavior was scarce or absent. Following VCS and interaction with lamb, maternal responses in the ovxSHORT group increased while social isolation responses decreased. Sexual receptivity occurred in non-pregnant ewes and in ovxSHORT group. Conclusion: some hormonal treatments +VCS can effectively induce maternal behavior and reduce social isolation responses. Long-term progestin treatment can inhibit postpartum estrus.

Male sexual behavior contributes to the maintenance of high LH pulsatility in anestrous female goats.

The objective of this study was to determine the importance of male sexual behavior in stimulating LH secretion in anovulatory female goats. Two groups of females (n=10 per group) were each exposed to a buck in sexual rest and submitted to natural daylength. In one group, the buck was awake, whereas in the other group, it was sedated to prevent its sexual behavior. Two other groups of goats (n=10 per group) were exposed to sexually active bucks that had been exposed to 2.5 months of long days. In one group, the buck was awake, and in the other group, it was sedated. LH secretion was determined every 15 min from 4 h before introducing the bucks to 8 h after, then every 15 min again from 20 to 24 h after introducing the bucks. The bucks submitted to natural daylength did not stimulate LH secretion (P>0.05), whether they were sedated or not. In contrast, both the awake and the sedated light-treated bucks induced an increase (P<0.05) of LH pulsatility in the first 4 h following their introduction. However, pulsatility remained elevated until 24 h in the females exposed to the light-treated awake buck, whereas in the group with the light-treated sedated buck, pulsatility diminished (P<0.05) after the first 4 h of stimulation by the buck. In conclusion, the sexual behavior of males contributes to the maintenance of a high LH pulsatility up to 24 h after introduction into a group of anovulatory goats.

Involvement of central muscarinic receptors in social and nonsocial learning in sheep.

Within 12 h following parturition, ewes develop visual and auditory recognition of their lamb. To investigate whether central cholinergic muscarinic transmission plays a specific role in this social learning, we studied the effects of a muscarinic antagonist on neonate recognition via visual/auditory cues and acquisition of a nonsocial visual discrimination. Injections of scopolamine (100 microg/kg; a muscarinic antagonist crossing the blood-brain barrier) after birth did not affect maternal behavior but impaired visual/auditory recognition of the offspring. Recognition impairment did not occur in mothers treated with methylscopolamine (100 microg/kg; a peripheral muscarinic antagonist), indicating that central muscarinic transmission is specifically involved in this social learning. Similar doses of scopolamine strongly delayed learning of a nonsocial, visual discrimination task in comparison to either control or methylscopolamine-treated ewes. Performance on this task was not affected when scopolamine treatment was applied after learning, demonstrating that central muscarinic receptors are necessary for acquisition but not for retrieval. These results suggest that the central muscarinic transmission participates in the establishment of visual/auditory recognition of conspecifics. Moreover, activation of central muscarinic receptors is critical for learning regardless of the social properties of the stimulus.

Different chemical fractions of fetal fluids account for their attractiveness at parturition and their repulsiveness during late-gestation in the ewe.

We have investigated whether the chemical components of fetal fluids (FFs), which elicit repulsion in late gestating ewes, are also those responsible for the attractiveness of fetal fluids at parturition. An aqueous fraction of FFs (A1), obtained after extraction with hexane, was tested for repulsion in late-pregnant ewes and for attraction at parturition. We also investigated if the repulsive and attractive characteristics of this A1 fraction were maintained after an additional extraction with dichloromethane (DCM, CH(2)Cl(2)) that produced two more fractions (aqueous/high polarity: A2 and dichloromethane/medium polarity: DCM). Thus, late-pregnant ewes were tested for repulsion of aqueous extracts of FFs (A1, A2 and DCM fractions) in a two-choice test of food preference, whereas parturient ewes were tested for attraction toward these same fractions in a two-choice test of licking warm spongy cloths. The A1 fraction was repulsive to late-pregnant ewes and attractive to parturient females. In contrast, neither the A2 nor the DCM fractions were repulsive to late-pregnant ewes, whereas both fractions were attractive to parturient ones. The discordance between the repulsive and attractive properties of the A2 and DCM fractions suggests that the attractiveness of FFs for parturient ewes and its repulsiveness for females outside the peripartum period depend on mixtures of substances that are at least partially different. Some compounds with high and medium polarity in the A2 and DCM fractions would act synergistically to generate the repulsiveness of FFs, whereas both high and medium polarity compounds can evoke attraction independently of each other.

Mother-young vocal communication and acoustic recognition promote preferential nursing in sheep.

In mammals with precocial neonates, exclusive maternal care and investment depend on mutual mother-young recognition. In sheep, this is ensured by rapid olfactory recognition of the neonate by its mother. However, recent studies suggest that other processes may participate in preferential maternal care. We investigated the possibility that acoustic communication promotes preferential nursing of the lamb. In the first of two studies, we examined the association between nursing and vocal activity in ewes and their lambs during the first 2 weeks of lactation. As early as 3 and 6 h postpartum, vocal activity was preferentially concentrated before nursing and by day 15 all vocal activity occurred within 2.5 min before nursing. In the second study, we tested the responses of ewes and lambs at 15 days postpartum to the playback of recorded bleats of their partner or from unrelated ewes and lambs. When playback was performed 30 min after a nursing episode, both ewes and lambs responded to bleats of their partner but not to bleats from alien subjects. When playback was performed 5 min after nursing, ewes did not respond to any lamb's bleats, while lambs continued responding to the bleats of their mothers, but significantly less than 30 min after nursing. Nursing therefore appears to play an important role in structuring very early vocal communication between the mother and her neonate. In turn, if the motivational state of the members of the mother-young dyad is adequate, this ensures the display of mutual acoustic recognition and prepares them for preferential nursing before maternal olfactory recognition of the lamb comes into play.

Maternal responsiveness and maternal selectivity in domestic sheep and goats: the two facets of maternal attachment.

Sheep and goats rapidly establish an exclusive relationship with their neonate following contact with it during a sensitive period of maternal responsiveness induced by the physiological events occurring at parturition. The data concerning the sensory, physiological, and neurobiological factors involved in the activation of both maternal responsiveness and the establishment of selective nursing indicates that these processes are activated simultaneously by the combined action of two main factors, the prepartum rise in circulating estrogen and the vaginocervical stimulation (VCS) caused by fetus expulsion. On the one hand, these two factors act on a neural network including the main olfactory system (MOB), the medial preoptic area (MPOA), and the paraventricular nucleus of the hypothalamus (PVN) to induce maternal responsiveness towards any neonate. The intracerebral release of oxytocin (OT) from the PVN, and the triggering of olfactory attraction for amniotic fluid (AF) are key elements in this process. On the other hand, VCS at birth also sets the MOB ready to memorize the individual odor of the neonate, through the release of peptides and neurotransmitters (noradrenaline and acetylcholine). In addition to the MOB, the network involved in recognition mainly includes the medial and cortical amygdala. Across consolidation processes, reorganization occurs in the network engaged in lamb recognition. Whether this memorization may be potentiated by other sensory cues is not known. The identification of the chemosensory compounds involved in the attraction for AF and in the recognition of the neonate is important for understanding the mechanisms of maternal attachment.

Establishment of vocal communication and discrimination between ewes and their lamb in the first two days after parturition.

In sheep, the mother-young relationship is characterized by an exclusive bond that relies on the rapid establishment of mutual recognition. However, the role of acoustic cues has been underestimated and neglected in this early discrimination. Therefore, the aim of the present study was (1) to characterize the vocal behavior of ewes and their lambs during the first 15 days postpartum and (2) to investigate the ability of newborn lambs and their mothers to discriminate each other from alien subjects within the first 48 hr postpartum on the basis of acoustic cues only. Ewes started to vocalize in the last 3 hr preceding parturition, emitting mainly high-pitched bleats. However, the bleating activity was highest during the first 3 hr following birth, consisting mainly of low-pitched bleats. Thereafter it decreased sharply until 24 hr postpartum, with most of the activity coming from the mother. In a two-choice test, ewes discriminated their own from an alien lamb on the basis of acoustic cues only at 24 hr postpartum, while lambs did so for their mother at 48 hr, which in both cases was much earlier than reported so far. We conclude that early vocal recognition between the ewe and her lamb may play an important role for the maintenance of mother-young contact.

Sensory and physiological determinants of maternal behavior in the goat (Capra hircus).

Maternal behavior in the goat appears at the time of parturition, partly under the activating influence of vaginocervical stimulation. Mothers actively lick their neonate and rapidly establish a selective bond with their kid through olfactory recognition. They also develop visual and acoustic recognition of the kid within 4 h following birth. Acoustic recognition is present at 48 h. The establishment of maternal recognition can be impaired by underfeeding during the second half of pregnancy. There is no indication that the mechanisms controlling the onset of maternal behavior and bonding are different from those reported in sheep, despite the fact that lambs start to follow their mother within a few hours after birth and kids hide for about a week. During lactation, the cues provided by the kid are necessary for the maintenance of maternal responsiveness, but suckling itself does not appear of primary importance. The presence of the kid also modulates the hormonal response to udder stimulation and influences recovery of postpartum sexual activity when kidding (i.e. birthing) takes place in autumn. Finally, the rapid establishment of mutual attachment between mother goats (does) and their kids offers the possibility to investigate an aspect of mother-young affiliation that is not present in many laboratory species.

From birth to colostrum: early steps leading to lamb survival.

New-born lambs have limited energy reserves and need a rapid access to colostrum to maintain homeothermy and survive. In addition to energy, colostrum provides immunoglobulins which ensure passive systemic immunity. Therefore, getting early access to the udder is essential for the neonate. The results from the literature reviewed here highlight the importance of the birth site as the location where the mutual bonding between the mother and her young takes place. Attraction to birth fluids by the periparturient ewe leads to intense licking of the lamb. Grooming not only dries, cleans and stimulates the newborn it also facilitates bonding through learning of its individual odour. Ewes having twins should ideally stay on the birth site for at least six hours in order to establish a strong bond with both lambs and favour lambs survival. However, primiparous ewes or ewes having high levels of emotivity are more likely to exhibit poor maternal behaviour. In addition, difficult parturition and weather conditions have an indirect effect on the behaviour of the mother and are other major causes of lamb death. On the lamb's side, rapid access to the udder and early suckling are extremely important. Delayed lactation or insufficient colostrum yield may be fatal especially since suckling has strong rewarding properties in the establishment of a preference for the mother, which in turn increases lamb survival. Insufficient access to the udder in mothers leaving the birth site too soon after parturition, especially in twin-bearing ewes, could also partly account for the high incidence of loss of mother-young contact and subsequent death in such lambs. Strategies to improve neonatal survival should be aimed at maximising lamb vigour, colostrum production, and mutual mother-young bonding through adequate feeding in late pregnancy and selection on behaviour.

Positive correlation between the body weight of anestrous goats and their response to the male effect with sexually active bucks.

In the present study, we analyzed the results of two years of response to the male effect in seasonally anestrous goats to investigate whether the activation of female reproductive activity by the male effect is related to the body weight of the females. Seventy-nine adult female Mexican mixed breed goats were used. The anestrous females were exposed during 15 days to sexually active males, and were classified into three categories according to their mean body weight +/-SD (42 +/- 9 kg) (Light: < or = 33 kg, n = 19; Medium: 34-50 kg, n = 46; Heavy: > or = 51 kg, n = 14). More than 98% of the goats from the Medium and Heavy groups showed at least one estrus behavior within the first 15 days following the introduction of the bucks, versus only 63% of the females from the Light group (P < 0.01). The interval between the introduction of the males and the onset of estrus behavior was longer in the females of the Light and Medium groups (4.2 +/- 0.8 and 3.3 +/- 0.3 days) than in the females of the Heavy group (2.0 +/- 0.2 days; P < 0.03). Also, body weight was negatively correlated with latency to first estrus (Spearman r = -0.57; P < 0.001). These results are in agreement with the hypothesis that the ability of anestrous goats to respond to the male effect is positively influenced by their body weight.

Mechanisms of activation of maternal behaviour in mammals.

In mammals, the activation of maternal behaviour depends on internal maternal factors related with parturition. The nature of these factors may vary between species, although oestradiol, foetus expulsion and intracerebral oxytocin are the most commonly encountered. They induce a period of specific receptivity to some sensory cues provided by the neonate. These cues (tactile, olfactory, auditory, ...) also vary between species. The interactions between the mother and her progeny during this period, known as the sensitive period, facilitate the maintenance of maternal responsiveness beyond the initial phase of activation by the factors internal to the mother. The ability of mothers to display a well-adapted maternal behaviour is also modulated by maternal experience gained at the first parturition. Furthermore, this capacity is also influenced by the nature of the interactions experienced in infancy with the own mother of the female, which can lead to a non-genomic transmission of some individual characteristics of maternal behaviour or sensitivity to stress across generations.

Genetic analysis of emotional reactivity in sheep: effects of the genotypes of the lambs and of their dams.

A total of 1347 weaned lambs from eight genotypes were tested over five consecutive years: Romanov (ROM) and Lacaune (LAC) pure breeds, the two F1 crossbreeds (RL and LR) and the offspring of ewes from these four genotypes sired with Berrichon-du-Cher rams (BCF). The lambs were individually exposed to three challenging tests involving novelty, human contact and social isolation. Ten synthetic variables were used to express social reactivity (i.e., active vs. passive strategy), exploratory activity and reactivity to humans. BCF crossbreds were more active (i.e., high bleats, locomotion and attempts to escape) than purebreds and F1. In contrast, ROM expressed more passive responses (i.e., low bleats and vigilance postures) than LAC and BCF crossbreds. In addition, ROM approached a motionless human less and had longer flight distances to an approaching human than did LAC and BCF crossbreds. When restrained, ROM, and to a lesser extent B x ROM and B x LR, avoided human contact more than did LAC, RL and B x LAC. Most of these differences were explained by direct additive genetic effects while maternal influences or heterosis effects were rarely significant. The highest heritability was for high bleats (h2 = 0.48). Females were more active and avoided human contact more than did males.

Prepartum peripherally-induced hyposmia does not reduce postpartum anoestrus duration in nursing goats.

Parturient goats rapidly develop exclusive nursing of their own litter that relies on olfactory recognition of the young. They also show a period of postpartum anoestrus whose duration depends on the presence of the kid. In cattle, maternal selectivity is one of the factors that delays the recovery of sexual activity. To investigate the possible influence of maternal selectivity on the duration of postpartum anoestrus in goats, we compared the recovery of estrus behavior by daily estrus detection with an active buck in intact and selective nursing goats (n = 24) with that of dams rendered non-selective by peripheral hyposmia with ZnSO4 (n = 18). Postpartum anoestrus duration was shorter in intact (68+/-7 days) than in hyposmic mothers (93+/-7 days; P < 0.05). However, the cycles of normal duration were less frequent in intact goats (P = 0.03). We conclude that in nursing goats, preventing the establishment of selective nursing by prepartum peripheral hyposmia does not reduce postpartum anoestrus duration. Our results suggest that daily exposure to the buck may result in an earlier recovery of ovarian activity in intact mothers.

Maternal experience influences the establishment of visual/auditory, but not olfactory recognition of the newborn lamb by ewes at parturition.

Maternal behavior in sheep is characterized by the rapid establishment of individual recognition of the lamb through the use of different sensory modalities. Olfactory recognition mediates acceptance at suckling whereas visual/auditory cues are involved in recognition from a distance. This study investigates (a) the timing of both types of recognition and (b) whether they can be influenced by maternal experience. Olfactory recognition was assessed at lambing, 30 min, 1, 2, or 4 hr postpartum by presenting successively an alien and the familiar lamb. Recognition at a distance was assessed at 6, 8, and 12 hr postpartum by using a two-choice test between the familiar and an alien lamb. A majority (33/51) of ewes showed selectivity at suckling as early as 30 min after parturition, and no differences due to maternal experience were observed. By contrast, in the two-choice recognition test, multiparous ewes showed a preference for their familiar lamb at 6 hr whereas primiparous mothers did so only after 24 hr of mother-young contact: The performance of biparous mothers was intermediate. Thus, both types of recognition can be rapidly established after parturition, and maternal experience has a differential effect on the dynamics of these learning processes.

Early recognition of newborn goat kids by their mother: I. Nonolfactory discrimination.

Mother sheep and goats develop an early bond with their neonate on the basis of olfactory recognition. We investigated whether goats were also able to show early (<24 hr postpartum) nonolfactory discrimination of their kids, as already reported in sheep. In a first experiment, we found that goats are not able to recognize their kid at 1 m away on the basis of olfactory cues alone. By contrast, they showed a significant preference for their own kid in a two-choice test as early as 4 hr postpartum, and prepartum maternal anosmia did not impede the ability of mothers to show discrimination. We conclude that goats, like sheep, are fully able to discriminate their neonate without the help of olfactory cues very early after parturition. The difference in the early spatial mother-young relationship between the two species due to the different behavior of the young (kids = hiders, lambs = followers), is not associated with marked differences in the dynamics or mechanisms controlling the development of recognition of the neonate by its mother.

Early recognition of newborn goat kids by their mother: II. Auditory recognition and evidence of an individual acoustic signature in the neonate.

The vocal recognition of newborn kids by their mother at 2 days postpartum and the possible existence of interindividual differences in the voice structure of newborn kids were investigated in two separate studies. The ability of goats to discriminate between the bleats of their own versus an alien kid was tested at 2 days postpartum in mothers being prevented access to visual and olfactory cues from the young. Goats spent significantly more time on the side of the enclosure from which their own kid was bleating, looked in its direction for longer, and responded more frequently to the bleats of their own than to those of the alien kid (p < 0.05). In the second study, the sonograms of 13 kids, studied from Days 1 to 5, showed significant interindividual differences for the five variables taken into account and on each of the 5 days (duration of bleat, fundamental frequency, peak frequency, and numbers of segments and of harmonics). The potential for individual coding ranged between 1.1 and 4.1, indicating that for some variables variations between individuals were greater than intraindividual variations. Furthermore, when considering the five parameters together, the discriminating scores showed an average of 95% in the 78 combinations of any 2 kids for any given day. Finally, some significant intraindividual differences also were found between days, suggesting ontogenic changes in the characteristics of the kid's voice in early life. Therefore, mother goats are likely to recognize the vocalizations of their 48-hr-old kids, as they show sufficient interindividual variability to allow the existence of individual vocal signatures, even though some of the characteristics of the bleats change rapidly over time.

Maternal olfaction differentially modulates oxytocin and prolactin release during suckling in goats.

In postparturient goats, olfactory recognition of the young allows the establishment of a selective bond between the mother and her kids. Once this bond is formed, the mother rejects alien young that attempt to suckle. We tested whether the development of the maternal selective bond in goats modulates prolactin (PRL) and oxytocin (OT) release in response to suckling. On day 37 of lactation, serial blood samples were taken during nursing of the mother's own or alien kid(s) in 10 intact/selective goats and in 10 goats rendered anosmic/nonselective through prepartum peripheral ZnSO(4) irrigation. Spontaneous nursing behavior was also studied weekly from day 7 to 30 of lactation, at which time milk production was measured. Maternal selectivity had no effect on PRL release, in contrast to OT release, which was significantly affected by this factor. Intact mothers released OT only when nursing their own kids, but not with aliens, while anosmic/nonselective dams showed an increase in OT levels regardless of the identity of the kids. In addition to these effects on maternal selectivity, the amplitude of the response of both hormones was lower in anosmic mothers than in intact mothers. Finally, nursing behavior and milk production were not significantly affected by anosmia. We conclude that maternal selective behavior in goats, which relies on the individual olfactory signature of the kid, modulates the OT, but not the PRL, response to suckling. In addition, perception of the smell of the young appears to have a general facilitatory effect, independent of the kid's identity, on the release of both hormones.