Configural memory of a blending aromatic mixture reflected in activation of the left orbital part of the inferior frontal gyrus.

Blending aromatic mixtures components naturally fuse to form a unique odor - a configuration- qualitatively different from each component's odor. Repeated exposure to the components either in the mixture or separately, favors respectively, configural and elemental processings. The neural bases of such processes are still unknown. We examined the brain correlates of the experienced-induced configural processing of a well-known model of binary blending odor mixture, the aromatic pineapple blending (AB, ethyl maltol + ethyl isobutyrate). Before fMRI recording, half of the participants were repeatedly exposed to the mixture (AB, group Gmix), with the other half exposed to its separate components (A and B; Gcomp). During the fMRI recording, all participants were stimulated with the mixture (AB) and the components (A and B). Finally, participants rated the number of odors perceived for each stimulus. Gmix perceived the AB mixture as less complex than did Gcomp. While Gcomp perceived the mixture as more complex than its components, Gmix did not. These results show the presence of experience-induced configural or elemental processing of the AB mixture in each group. Contrasting the brain activity of Gcomp and Gmix, when stimulated with AB, revealed higher activation in the left orbital part of the inferior frontal gyrus. This result sheds light on this area's function, commonly found activated in olfactory studies, and closely connected with the lateral orbitofrontal cortex. We discuss the role of this area as a mediator of configural percepts between temporal and orbitofrontal areas involved in configural memory processes.

Insights into suckling rabbit feeding behaviour: acceptability of different creep feed presentations and attractiveness for sensory feed additives.

In young rabbit, digestive disorders are frequently observed around weaning. Stimulating the onset of feed intake in the suckling rabbit might be a way to promote gut health. The aim of this study was to determine the rabbit's acceptability for different feed presentations and its preferences for flavours at an early stage of life. Two trials were conducted to evaluate the effects of physical form and flavouring on creep feed attractiveness. All the diets tested were provided in the nest from 3 to 17 days, and the daily intake per litter was recorded as of 8 days of age. In the first trial, five feed presentations were tested separately (n = 60 litters). Three dry presentations were chosen: commercial pellet (P), crumb from commercial pellet (cP) and crumb from beet pulp pellet (cBP). Hydrated feeds were also provided with either raw fodder beetroot (B) or a semi-solid feed in agar gel form produced with fodder beetroot juice and pulp (gB). In the second trial, double-choice tests were performed on four feed gels (n = 72 litters), leading to six comparison treatments. These agar gels were made of pellet mash without or with a sensory additive: one non-odorised control gel and three gels with 0.20% banana flavour, 0.06% red berry flavour and 0.10% vanilla flavour, respectively. In the first trial, kits ate more gB in fresh matter than other feed presentations (P < 0.001), with a total intake of 7.0 ± 1.8 g/rabbit from 8 to 17 days. In DM, the total consumption of pellets P (1.6 ± 0.4 g of DM/rabbit) was the highest together with the gB form (1.4 ± 0.4 g of DM/rabbit), whereas cBP was barely consumed (0.3 ± 0.1 g of DM/rabbit). Gel feed supplemented with vanilla was slightly more consumed than other flavoured and non-odorised gels (relative consumption of 57% when compared to control gel; P = 0.001). The gel feed intake was independent of the milk intake but was correlated with the litter weight at 3 days (r = 0.40, P < 0.001). In both trials, rabbit growth before and after weaning was not affected by the type of creep feed provided. Our results confirmed that providing creep feed promotes the solid intake of rabbits at early stages. Gel feed form motivated rabbits to eat and vanilla flavour supplementation increased the feed palatability. Those creep feed characteristics should be explored further for seeking effective stimulation of the onset of the feed intake in suckling rabbit.

Pheromone-induced odor learning modifies Fos expression in the newborn rabbit brain.

Associative learning contributes crucially to adjust the behavior of neonates to the permanently changing environment. In the European rabbit, the mammary pheromone (MP) excreted in milk triggers sucking behavior in newborns, and additionally promotes very rapid learning of initially neutral odor cues. Such stimuli become then as active as the MP itself to elicit the orocephalic motor responses involved in suckling. In this context, the rabbit is an interesting model to address the question of brain circuits early engaged by learning and memory. Here, we evaluated the brain activation (olfactory bulb and central regions) induced in 4-day-old pups by an odorant (ethyl acetoacetate, EAA) after single pairing with the MP and its subsequent acquired ability to elicit sucking-related behavior (conditioned group) or after mere exposure to EAA alone (unconditioned group). The brain-wide mapping of c-Fos expression was used to compare neural activation patterns in both groups. Evidence of high immunostaining to odorant EAA occurred in the mitral+granule cells layer of the main olfactory bulb in pups previously exposed to EAA in association with the MP. These pups also showed higher expression of Fos in the piriform cortex, the hypothalamic lateral preoptic area and the amygdala (cortical and basal nuclei). Thus, MP-induced odor learning induces rapid brain modifications in rabbit neonates. The cerebral framework supporting the acquisition appears however different compared to the circuit involved in the processing of the MP itself.

Brain processing of the mammary pheromone in newborn rabbits.

Chemosignals strongly contribute to social interactions in mammals, including mother-young relationships. In the European rabbit, a volatile compound emitted by lactating females in milk, the 2-methylbut-2-enal, has been isolated. Carrying the properties of a pheromone, in particular the spontaneous ability to release critical sucking-related movements in newborns, it has been called the mammary pheromone (MP). Lesion of the vomeronasal organ and preliminary 2-deoxyglucose data suggested that the MP could be processed by the main olfactory system. However, the neuronal substrate that sustains the MP-induced response of neonates remained unknown. Here, we evaluated Fos expression in 4-day-old-rabbits exposed to the MP (in comparison with control neonates exposed to non-relevant odorant, no odorant or unmanipulated pups) both at the level of the olfactory bulb and central brain regions. Evidence of high and widespread Fos immunoreactivity in the main olfactory bulb appear in MP pups while the accessory olfactory bulb exhibits a negligible staining. However, no obvious bulbar pattern of Fos expression is observed, when in contrast a certain pattern emerges with the neutral odorant. Compared to this latter, the MP exposure increases Fos expression in the anterior piriform cortex, the organum vasculosum of the lamina terminalis and the habenula, with a tendency in the lateral preoptic region. For the first time, a pheromone essential for mother-young interaction is thus highlighted for its processing by the main olfactory system, the whole olfactory bulb, and by brain regions involved in osmoregulation, thirst and motivation-guided motor responses.

Perceptual blending in odor mixtures depends on the nature of odorants and human olfactory expertise.

Our olfactory system is confronted with complex mixtures of odorants, often recognized as single entities due to odor blending (e.g., coffee). In contrast, we are also able to discriminate odors from complex mixtures (e.g., off-odors). Therefore, the olfactory system is able to engage either configural or elemental processes when confronted with mixtures. However, the rules that govern the involvement of these processes during odor perception remain poorly understood. In our first experiment, we examined whether simple odorant mixtures (binary/ternary) could elicit configural perception. Twenty untrained subjects were asked to evaluate the odor typicality of mixtures and their constituents. The results revealed a significant increase in odor typicality in some but not all mixtures as compared with the single components, which suggest that perceptual odor blending can occur only in specific mixtures (configural processing). In our second experiment, we tested the hypothesis that general olfactory expertise can improve elemental perception of mixtures. Thirty-two trained subjects evaluated the odor typicality of the stimuli presented during the first experiment, and their responses were compared with those obtained from the untrained panelists. The results support the idea that general training with odors increases the elemental perception of binary and ternary blending mixtures.

The reactivity of neonatal rabbits to the mammary pheromone as a probe for viability.

Newborn rabbits depend on a daily nursing interaction with the mother to gain milk and to survive. During this interaction, they localise and seize the nipples displaying a typical behaviour triggered by maternal odour cues. The mammary pheromone constitutes such a signal in domestic rabbits: it elicits sucking-related movements in more than 90% of the pups. However, some newborns remain unresponsive to the presentation of the pheromone, even pups apparently healthy and highly motivated to suck. The main goal of the present study was therefore to explore the link between the unresponsiveness of rabbit pups to the mammary pheromone and their growth and survival in breeding conditions. To that end, 293 newborns from 30 litters were tested for their head searching-oral grasping responses to the mammary pheromone on days 1 and 3, and their milk intake and mortality were followed up from days 1 to 21. It was hypothesised that unresponsive newborns would have subsequent difficulties in finding nipples, sucking and surviving. Early weight and success in milk intake were further considered as mediating factors in growth and viability. The results showed that pups that were unresponsive to the mammary pheromone on day 1 were less successful in gaining milk and had a higher rate of mortality than the responsive pups. However, this impact was modulated by the weight of pups: it appeared only in the lightest newborns. Moreover, this impact vanished on day 3. On the other hand, the pup weight and sucking success on days 1 to 3 strongly influenced viability and growth during the period extending from days 1 to 21. Taken together, the results show that the day-1 responsiveness of rabbit pups to the mammary pheromone can be considered as an indicator of individual viability in pups having a small weight (<48 g on day 1). The predictive validity of the pups' pheromonal reactivity seems however time-limited as it works only during the first, but crucial, postnatal days.

Attraction of newborn rabbits to abdominal odors of adult conspecifics differing in sex and physiological state.

The present study aimed at assessing the ability of newborn rabbits (1-3 days) to detect and discriminate abdominal odors emitted by adult conspecifics varying in sex and physiological state. Pups were submitted to a two-choice test exclusively based on olfaction, which permitted exposure to conspecifics either during successive or simultaneous presentations. Their orientation toward either of the two stimuli was timed. These behavioral assays revealed that (a) pups display attraction to odors of the abdomen of nonlactating, nonpregnant females and males when pitted against a control stimulus; (b) pups preferred the odor of nonlactating, nonpregnant females when simultaneously presented with the odor of a male; (c) all sexually mature females (virgin, pregnant, or lactating) were attractive to pups; (d) the odor of lactating females was preferred to the odor of either nonlactating, nonpregnant or pregnant females. Thus, from the first day after birth, newborn rabbits are able to olfactorily discriminate conspecifics from cues emitted at the abdominal level and display the strongest attraction to lactating females.

Immediate postnatal sucking in the rabbit: its influence on pup survival and growth.

This study was aimed at characterising the impact of immediate postnatal sucking on pup survival and development. The interactive effects of postnatal success with the day 0 weight of pups, the nest-access regimen (controlled or free) or parity of females was investigated. Pups (n = 900) were categorised according to their initial ingestion of colostrum. In primiparous does: (1) pup mortality between d0-d10 was higher for unsuccessful than for successful early suckers; (2) lighter d0-weight reduced survival for unsuccessful but not for successful pups; (3) free nest-access of females annihilated the survival advantage fostered by the initial sucking success. In secondiparous does, these impacts waned. Finally, whatever the does' parity, only d0-weight influenced pup weight-gain between d0-21. Thus, pup survival seemed to depend (at least in primiparae) on their ability to suck right after birth, and to display a pattern of energy saving without being disturbed by the females' nest entries.