Crocodile perception of distress in hominid baby cries.

It is generally argued that distress vocalizations, a common modality for alerting conspecifics across a wide range of terrestrial vertebrates, share acoustic features that allow heterospecific communication. Yet studies suggest that the acoustic traits used to decode distress may vary between species, leading to decoding errors. Here we found through playback experiments that Nile crocodiles are attracted to infant hominid cries (bonobo, chimpanzee and human), and that the intensity of crocodile response depends critically on a set of specific acoustic features (mainly deterministic chaos, harmonicity and spectral prominences). Our results suggest that crocodiles are sensitive to the degree of distress encoded in the vocalizations of phylogenetically very distant vertebrates. A comparison of these results with those obtained with human subjects confronted with the same stimuli further indicates that crocodiles and humans use different acoustic criteria to assess the distress encoded in infant cries. Interestingly, the acoustic features driving crocodile reaction are likely to be more reliable markers of distress than those used by humans. These results highlight that the acoustic features encoding information in vertebrate sound signals are not necessarily identical across species.

Higher-order trace conditioning in newborn rabbits.

Temporal contingency is a key factor in associative learning but remains weakly investigated early in life. Few data suggest simultaneous presentation is required for young to associate different stimuli, whereas adults can learn them sequentially. Here, we investigated the ability of newborn rabbits to perform sensory preconditioning and second-order conditioning using trace intervals between odor presentations. Strikingly, pups are able to associate odor stimuli with 10- and 30-sec intervals in sensory preconditioning and second-order conditioning, respectively. The effectiveness of higher-order trace conditioning in newborn rabbits reveals that very young animals can display complex learning despite their relative immaturity.

Biological constraints on configural odour mixture perception.

Animals, including humans, detect odours and use this information to behave efficiently in the environment. Frequently, odours consist of complex mixtures of odorants rather than single odorants, and mixtures are often perceived as configural wholes, i.e. as odour objects (e.g. food, partners). The biological rules governing this 'configural perception' (as opposed to the elemental perception of mixtures through their components) remain weakly understood. Here, we first review examples of configural mixture processing in diverse species involving species-specific biological signals. Then, we present the original hypothesis that at least certain mixtures can be processed configurally across species. Indeed, experiments conducted in human adults, newborn rabbits and, more recently, in rodents and honeybees show that these species process some mixtures in a remarkably similar fashion. Strikingly, a mixture AB (A, ethyl isobutyrate; B, ethyl maltol) induces configural processing in humans, who perceive a mixture odour quality (pineapple) distinct from the component qualities (A, strawberry; B, caramel). The same mixture is weakly configurally processed in rabbit neonates, which perceive a particular odour for the mixture in addition to the component odours. Mice and honeybees also perceive the AB mixture configurally, as they respond differently to the mixture compared with its components. Based on these results and others, including neurophysiological approaches, we propose that certain mixtures are convergently perceived across various species of vertebrates/invertebrates, possibly as a result of a similar anatomical organization of their olfactory systems and the common necessity to simplify the environment's chemical complexity in order to display adaptive behaviours.

Configural perception of a binary olfactory mixture in honey bees, as in humans, rodents and newborn rabbits.

How animals perceive and learn complex stimuli, such as mixtures of odorants, is a difficult problem, for which the definition of general rules across the animal kingdom remains elusive. Recent experiments conducted in human and rodent adults as well as newborn rabbits suggested that these species process particular odor mixtures in a similar, configural manner. Thus, the binary mixture of ethyl isobutyrate (EI) and ethyl maltol (EM) induces configural processing in humans, who perceive a mixture odor quality (pineapple) that is distinct from the quality of each component (strawberry and caramel). Similarly, rabbit neonates treat the mixture differently, at least in part, from its components. In the present study, we asked whether the properties of the EI.EM mixture extend to an influential invertebrate model, the honey bee Apis mellifera. We used appetitive conditioning of the proboscis extension response to evaluate how bees perceive the EI.EM mixture. In a first experiment, we measured perceptual similarity between this mixture and its components in a generalization protocol. In a second experiment, we measured the ability of bees to differentiate between the mixture and both of its components in a negative patterning protocol. In each experimental series, the performance of bees with this mixture was compared with that obtained with four other mixtures, chosen from previous work in humans, newborn rabbits and bees. Our results suggest that when having to differentiate mixture and components, bees treat the EI.EM in a robust configural manner, similarly to mammals, suggesting the existence of common perceptual rules across the animal kindgdom.

Developmental changes in elemental and configural perception of odor mixtures in young rabbits.

Development can change the way organisms represent their environment and affect their behavior. In vision, complex stimuli are treated as the sum of their elements (elemental perception) in children or as a whole (configural perception) in adults. However, the influence of development in elemental/configural perception has never been tested in olfaction. Here we explored this issue in young rabbits, which are known to perceive during the neonatal period certain binary odor mixtures elementally and others weak configurally. Using conditioning and behavioral testing procedures, we set out six experiments evaluating the putative evolution of their odor perception between birth and weaning. Results highlighted that between postnatal days 2 and 9 the perception of an initially weak configural mixture became robust configural while that of two elemental mixtures did not. Additional switches from elemental to configural perception were observed at postnatal day 24. The use of a chemically more complex senary mixture resulted also in a shift from weak to robust configural perception between postnatal days 2 and 9. Thus, the perception of certain odor mixtures may rapidly evolve toward a more holistic mode in young rabbits, which may help simplifying their representation of the environment once out of the nest.

Brain anatomy of the 4-day-old European rabbit.

The European rabbit (Oryctolagus cuniculus) is a widely used model in fundamental, medical and veterinary neurosciences. Besides investigations in adults, rabbit pups are relevant to study perinatal neurodevelopment and early behaviour. To date, the rabbit is also the only species in which a pheromone - the mammary pheromone (MP) - emitted by lactating females and active on neonatal adaptation has been described. The MP is crucial since it contributes directly to nipple localisation and oral seizing in neonates, i.e. to their sucking success. It may also be one of the non-photic cues arising from the mother, which stimulates synchronisation of the circadian system during pre-visual developmental stages. Finally, the MP promotes neonatal odour associative and appetitive conditioning in a remarkably rapid and efficient way. For these different reasons, the rabbit offers a currently unique opportunity to determine pheromonal-induced brain processing supporting adaptation early in life. Therefore, it is of interest to create a reference work of the newborn rabbit pup brain, which may constitute a tool for future multi-disciplinary and multi-approach research in this model, and allow comparisons related to the neuroethological basis of social and feeding behaviour among newborns of various species. Here, in line with existing experimental studies, and based on original observations, we propose a functional anatomical description of brain sections in 4-day-old rabbits with a particular focus on seven brain regions which appear important for neonatal perception of sensory signals emitted by the mother, circadian adaptation to the short and single daily nursing of the mother in the nest, and expression of specific motor actions involved in nipple localisation and milk intake. These brain regions involve olfactory circuits, limbic-related areas important in reward, motivation, learning and memory formation, homeostatic areas engaged in food anticipation, and regions implicated in circadian rhythm and arousal, as well as in motricity.

Odorant Metabolism Analysis by an Automated Ex Vivo Headspace Gas-Chromatography Method.

In the olfactory epithelium (OE), odorant metabolizing enzymes have the dual function of volatile component detoxification and active clearance of odorants from the perireceptor environment to respectively maintain the integrity of the tissues and the sensitivity of the detection. Although emphasized by recent studies, this enzymatic mechanism is poorly documented in mammals. Thus, olfactory metabolism has been characterized mainly in vitro and for a limited number of odorants. The automated ex vivo headspace gas-chromatography method that was developed here was validated to account for odorant olfactory metabolism. This method easily permits the measurement of the fate of an odorant in the OE environment, taking into account the odorant gaseous state and the cellular structure of the tissue, under experimental conditions close to physiological conditions and with a high reproducibility. We confirmed here our previous results showing that a high olfactory metabolizing activity of the mammary pheromone may be necessary to maintain a high level of sensitivity toward this molecule, which is critical for newborn rabbit survival. More generally, the method that is presented here may permit the screening of odorants metabolism alone or in mixture or studying the impact of aging, pathology, polymorphism or inhibitors on odorant metabolism.

Neonatal representation of odour objects: distinct memories of the whole and its parts.

Extraction of relevant information from highly complex environments is a prerequisite to survival. Within odour mixtures, such information is contained in the odours of specific elements or in the mixture configuration perceived as a whole unique odour. For instance, an AB mixture of the element A (ethyl isobutyrate) and the element B (ethyl maltol) generates a configural AB percept in humans and apparently in another species, the rabbit. Here, we examined whether the memory of such a configuration is distinct from the memory of the individual odorants. Taking advantage of the newborn rabbit's ability to learn odour mixtures, we combined behavioural and pharmacological tools to specifically eliminate elemental memory of A and B after conditioning to the AB mixture and evaluate consequences on configural memory of AB. The amnesic treatment suppressed responsiveness to A and B but not to AB. Two other experiments confirmed the specific perception and particular memory of the AB mixture. These data demonstrate the existence of configurations in certain odour mixtures and their representation as unique objects: after learning, animals form a configural memory of these mixtures, which coexists with, but is relatively dissociated from, memory of their elements. This capability emerges very early in life.

When the nose must remain responsive: glutathione conjugation of the mammary pheromone in the newborn rabbit.

In insects, xenobiotic-metabolizing enzymes were demonstrated to regulate pheromones inactivation, clearing them from the olfactory periphery and keeping receptors ready for stimulation renewal. Here, we investigate whether similar processes could occur in mammals, focusing on the pheromonal communication between female rabbits and their newborns. Lactating rabbits emit in their milk a volatile aldehyde, 2-methylbut-2-enal, that elicits searching-grasping in neonates; called the mammary pheromone (MP), it is critical for pups which are constrained to find nipples within the 5 min of daily nursing. For newborns, it is thus essential to remain sensitive to this odorant during the whole nursing period to display several actions of sucking. Here, we show that the MP is enzymatically conjugated to glutathione in newborn olfactory epithelium (OE), in accordance with the high mRNA expression of glutathione transferases evidenced by quantitative reverse transcription-PCR. This activity in the nose is higher than in the liver and in OE of newborns compared with weanlings (no more responsive to the pheromone). Therefore, the results pinpoint the existence of a high level of MP-glutathione conjugation activity in the OE of young rabbits, especially in the developmental window where the perceptual sensitivity toward the MP is crucial for survival.

Sensory preconditioning in newborn rabbits: from common to distinct odor memories.

This study evaluated whether olfactory preconditioning is functional in newborn rabbits and based on joined or independent memory of odorants. First, after exposure to odorants A+B, the conditioning of A led to high responsiveness to odorant B. Second, responsiveness to B persisted after amnesia of A. Third, preconditioning was also functional with two overlapping pairs of odorants (A+B and B+C) and amnesia of one odorant did not affect memory of the others. Thus, incidental pairing of odorants allows reinforcement of one odorant to implicitly reinforce the others, the bond then vanishes, and the memory of each element becomes independent.

Dynamic developmental changes in neurotransmitters supporting infant attachment learning.

Infant survival relies on rapid identification, remembering and behavioral responsiveness to caregivers' sensory cues. While neural circuits supporting infant attachment learning have largely remained elusive in children, use of invasive techniques has uncovered some of its features in rodents. During a 10-day sensitive period from birth, newborn rodents associate maternal odors with maternal pleasant or noxious thermo-tactile stimulation, which gives rise to a preference and approach behavior towards these odors, and blockade of avoidance learning. Here we review the neural circuitry supporting this neonatal odor learning, unique compared to adults, focusing specifically on the early roles of neurotransmitters such as glutamate, GABA (Gamma-AminoButyric Acid), serotonin, dopamine and norepinephrine, in the olfactory bulb, the anterior piriform cortex and amygdala. The review highlights the importance of deepening our knowledge of age-specific infant brain neurotransmitters and behavioral functioning that can be translated to improve the well-being of children during typical development and aid in treatment during atypical development in childhood clinical practice, and the care during rearing of domestic animals.

Independence of first- and second-order memories in newborn rabbits.

The mammary pheromone promotes the acquisition of novel odorants (CS1) in newborn rabbits. Here, experiments pinpoint that CS1 becomes able to support neonatal learning of other odorants (CS2). We therefore evaluated whether these first- and second-order memories remained dependent after reactivation. Amnesia induced after CS2 recall selectively blocked this memory, when recall and amnesia of CS1 left the souvenir of CS2 safe; this finding partially differed from results obtained in adult mammals. Thus, in this model of neonatal appetitive odor learning, second-order memory seems to depend on first-order memory for its formation but not for its maintenance.

Proportion of odorants impacts the configural versus elemental perception of a binary blending mixture in newborn rabbits.

Processing of odor mixtures by neonates is weakly understood. Previous studies showed that a binary mixture of ethyl isobutyrate/ethyl maltol (odorants A/B) blends in newborn rabbits at the 30/70 ratio: Pups would perceive a configural odor in addition to the components' odors. Here, we investigated whether the emergence of this additional odor in AB is determined by specific ratio(s) of A and B. To that goal, we tested whether pups discriminated between AB mixtures with lower (A(-)B, 8/92 ratio) or higher (A(+)B, 68/32) proportion of A. In Experiment 1, pups conditioned to A (or B) responded to A(-)B and A(+)B but not to AB. In Experiment 2, pups responded to A(-)B after learning of A(-) (and to A(+)B after learning of A(+)) but not to AB. In Experiment 3, after conditioning to A(-)B pups responded to A(-) and B (and to A(+) and B after learning of A(+)B) but not or less to AB. In Experiment 4, pups responded to A(-)B and A(+)B after conditioning to AB. These results confirm the configural perception of certain odor mixtures by young organisms and reveal that the proportion of components is a key factor influencing their coding, recognition, and discrimination of complex stimuli.

Experience influences elemental and configural perception of certain binary odour mixtures in newborn rabbits.

Elemental and configural olfactory perception allows interaction with the environment from very early in life. To evaluate how newborn rabbits can extract and respond to information from the highly complex chemical surroundings, and how experience acts on this sensory, cognitive and behavioural capability, we ran a study in four steps including a total of eight experiments. We mainly used a binary AB mixture comprising ethyl isobutyrate (component A) and ethyl maltol (component B), previously shown as a bearer of blending properties; in rabbit pups (as in human adults), the mixture elicits a weak configural perception, i.e. the perception of a configural odour different from the odours of the components. First, a repeated exposure to one component of AB led to a more elemental perception of this mixture; conversely, a repeated exposure to AB facilitated its configural processing. Second, similar impact of experience did not appear with a non-blending AC mixture (ethyl isobutyrate-guaïacol). Third, repeated exposure to AB impacted not only the perception of AB, but also and in the same way the perception of the AC mixture sharing one component, and reciprocally. However, facilitation to perceive one mixture in one mode (configural/elemental) was not generalized to a mixture sharing no components with the experienced mixture [AB versus DE (damascenone and vanillin)]. Thus, experience contributes to the neonatal perception of odour mixtures and adds plasticity to the perceptual system. However, this impact remains dependent on the chemical composition of the mixtures.

Odor discrimination in terrestrial and aquatic environments in California sea lions (Zalophus californianus) living in captivity.

Pinnipeds, as any mammal species, use multimodal signals, including olfactory ones, to ensure vital functions. Thus, some pinniped species seem able to use olfaction in both social and foraging contexts and to discriminate between different odors in air including both natural and artificial odors, but studies on that topic remain scarce. Here, we studied the olfactory capabilities of California sea lions living in captivity at La Flèche Zoo (France) in both terrestrial and aquatic environments. We used two categories of odors: social odors (from familiar individuals of the same group, unfamiliar individuals from another Zoo, animal zookeepers and a terrestrial carnivore) and non-social odors (food and odors identified as repellents in certain vertebrates). Several behavioral parameters were measured and analyzed as the number and duration of contact with the odor, mouth openings, vocalizations (air only) and air bubble production (water only). Our results, although limited by the low number of animals monitored (n = 5), suggest that California sea lions are able to discriminate between different odors both in the air and under water. In the aquatic environment, the process allowing the perception of odors remains to be characterized. Applications to this work could be considered in captive conditions as well as in the wild.

Brief olfactory learning drives perceptive sensitivity in newborn rabbits: New insights in peripheral processing of odor mixtures and induction.

Perception of the wide, complex and moving odor world requires that the olfactory system engages processing mechanisms ensuring detection, discrimination and environment adaptation, as early as the peripheral stages. Odor items are mainly elicited by odorant mixtures which give rise to either elemental or configural perceptions. Here, we first explored the contribution of the peripheral olfactory system to configural and elemental perception through odorant interactions at the olfactory receptor (OR) level. This was done in newborn rabbits, which offer the opportunity to pair peripheral electrophysiology and well characterized behavioral responses to two binary mixtures, AB and A'B', which differ in their component ratio (A: ethyl isobutyrate, B: ethyl maltol), and that rabbit pups respectively perceived configurally and elementally. Second, we studied the influence on peripheral reactivity of the brief but powerful learning of one mixture component (odorant B), conditioned by association with the mammary pheromone (MP), which allowed us to assess the possible implication of the phenomenon called induction in neonatal odor learning. Induction is a plasticity mechanism expected to alter both the peripheral electrophysiological responses to, and perceptual detection threshold of, the conditioned stimulus. The results reveal that perceptual modes are partly rooted in differential peripheral processes, the AB configurally perceived mixture mirroring odorant antagonist interactions at OR level to a lesser extent than the A'B' elementally perceived mixture. Further, the results highlight that a single and brief MP-induced odor learning episode is sufficient to alter peripheral responses to the conditioned stimulus and mixtures including it, and shifts the conditioned stimulus detection threshold towards lower concentrations. Thus, MP-induced odor learning relies on induction phenomenon in newborn rabbits.

A pheromone to behave, a pheromone to learn: the rabbit mammary pheromone.

Birth is part of a continuum and is a major developmental change. Newborns need to adapt rapidly to the environment in terms of physiology and behaviour, and ability to locate the maternal source of milk is vital. Mechanisms have evolved resulting in the emission of olfactory cues by the mother and the processing of these cues by the young. Here, we focus on some sensory, cognitive and behavioural strategies developed by the European rabbit (Oryctolagus cuniculus) that optimize the early development of offspring. In this species, chemosensory communication between the mother and young plays a critical role in eliciting adaptive neonatal responses. In particular, lactating females release a molecule, the mammary pheromone, which has several functional impacts. It triggers orocephalic responses involved in the quick localization of nipples and sucking. Moreover, this unconditioned signal promotes rapid appetitive learning of novel odorants, acting as a potent organizer of neonatal cognition. The mammary-pheromone-induced odour memory requires consolidation/reconsolidation processes to be maintained in the long term. Finally, as this mode of conditioning also promotes learning of mixtures of odorants, it supports investigations related to the capacity of neonatal olfaction to extract biological value from the complex environment.

Chemical and behavioural characterization of the rabbit mammary pheromone.

Mammals owe part of their evolutionary success to the harmonious exchanges of information, energy and immunity between females and their offspring. This functional reciprocity is vital for the survival and normal development of infants, and for the inclusive fitness of parents. It is best seen in the intense exchanges taking place around the mother's offering of, and the infant's quest for, milk. All mammalian females have evolved behavioural and sensory methods of stimulating and guiding their inexperienced newborns to their mammae, whereas newborns have coevolved means to respond to them efficiently. Among these cues, maternal odours have repeatedly been shown to be involved, but the chemical identity and pheromonal nature of these cues have not been definitively characterized until now. Here we focus on the nature of an odour signal emitted by the female rabbit to which newborn pups respond by attraction and oral grasping, and provide a complete chemical and behavioural description of a pheromone of mammary origin in a mammalian species.

Pheromone-induced olfactory memory in newborn rabbits: Involvement of consolidation and reconsolidation processes.

Mammary pheromone (MP)-induced odor memory is a new model of appetitive memory functioning early in a mammal, the newborn rabbit. Some properties of this associative memory are analyzed by the use of anisomycin as an amnesic agent. Long-term memory (LTM) was impaired by anisomycin delivered immediately, but not 4 h after either acquisition or reactivation. Thus, the results suggest that this form of neonatal memory requires both consolidation and reconsolidation. By extending these notions to appetitive memory, the results reveal that consolidation and reconsolidation processes are characteristics of associative memories of positive events not only in the adult, but also in the newborn.