Odor-driven face-like categorization in the human infant brain.

Understanding how the young infant brain starts to categorize the flurry of ambiguous sensory inputs coming in from its complex environment is of primary scientific interest. Here, we test the hypothesis that senses other than vision play a key role in initiating complex visual categorizations in 20 4-mo-old infants exposed either to a baseline odor or to their mother's odor while their electroencephalogram (EEG) is recorded. Various natural images of objects are presented at a 6-Hz rate (six images/second), with face-like object configurations of the same object categories (i.e., eliciting face pareidolia in adults) interleaved every sixth stimulus (i.e., 1 Hz). In the baseline odor context, a weak neural categorization response to face-like stimuli appears at 1 Hz in the EEG frequency spectrum over bilateral occipitotemporal regions. Critically, this face-like-selective response is magnified and becomes right lateralized in the presence of maternal body odor. This reveals that nonvisual cues systematically associated with human faces in the infant's experience shape the interpretation of face-like configurations as faces in the right hemisphere, dominant for face categorization. At the individual level, this intersensory influence is particularly effective when there is no trace of face-like categorization in the baseline odor context. These observations provide evidence for the early tuning of face-(like)-selective activity from multisensory inputs in the developing brain, suggesting that perceptual development integrates information across the senses for efficient category acquisition, with early maturing systems such as olfaction driving the acquisition of categories in later-developing systems such as vision.

Testing detectability, attractivity, hedonic specificity, extractability, and robustness of colostrum odor-Toward an olfactory bioassay for human neonates.

Human milk odor is attractive and appetitive for human newborns. Here, we studied behavioral and heart-rate (HR) responses of 2-day-old neonates to the odor of human colostrum. To evaluate detection in two conditions of stimulus delivery, we first presented the odor of total colostrum against water. Second, the hedonic specificity of total colostrum odor was tested against vanilla odor. Third, we delivered only the fresh effluvium of colostrum separated from the colostrum matrix; the stability of this colostrum effluvium was then tested after deep congelation; finally, after sorptive extraction of fresh colostrum headspace, we assessed the activity of colostrum volatiles eluting from the gas chromatograph (GC). Regardless of the stimulus-delivery method, neonates displayed attraction reactions (HR decrease) as well as appetitive oral responses to the odor of total colostrum but not to vanilla odor. The effluvium separated from the fresh colostrum matrix remained appetitive but appeared labile under deep freezing. Finally, volatiles from fresh colostrum effluvium remained behaviorally active after GC elution, although at lower magnitude. In sum, fresh colostrum effluvium and its eluate elicited a consistent increase in newborns' oral activity (relative to water or vanilla), and they induced shallow HR decrease. Newborns' appetitive oral behavior was the most reproducible response criterion to the effluvium of colostrum. In conclusion, a set of unidentified volatile compounds from human colostrum is robust enough after extraction from the original matrix and chromatographic processing to continue eliciting appetitive responses in neonates, thus opening new directions to isolate and assay specific volatile molecules of colostrum.

Flavor Sensing in Utero and Emerging Discriminative Behaviors in the Human Fetus.

The diet of pregnant women exposes fetuses to a variety of flavors consisting of compound sensations involving smell, taste, and chemesthesis. The effects of such prenatal flavor exposure on chemosensory development have so far been measured only postnatally in human infants. Here, we report the first direct evidence of human fetal responsiveness to flavors transferred via maternal consumption of a single-dose capsule by measuring frame-by-frame fetal facial movements. Pregnant women and their fetuses based in the northeast of England were involved in this study from 32 to 36 weeks' gestation. Fetuses exposed to carrot flavor (n = 35) showed "lip-corner puller" and "laughter-face gestalt" more frequently, whereas fetuses exposed to kale flavor (n = 34) showed more "upper-lip raiser," "lower-lip depressor," "lip stretch," "lip presser," and "cry-face gestalt" in comparison with the carrot group and a control group not exposed to any flavors (n = 30). The complexity of facial gestalts increased from 32 to 36 weeks in the kale condition, but not in the carrot condition. Findings of this study have important implications for understanding the earliest evidence for fetal abilities to sense and discriminate different flavors.

Comparative Investigation of Conventional and Innovative Headspace Extraction Methods to Explore the Volatile Content of Human Milk.

The odor of human milk induces search-like movements and oral activation in newborns, which increases their chances of taking advantage of milk intake and benefits. However, the underlying volatile fraction of human milk remains understudied. This study aimed to devise a simple method to extract a wide range of volatile compounds from small-volume human milk samples. Headspace solid phase micro-extraction (HS-SPME) with a Car/PDMS fiber and dynamic headspace extraction (D-HS) with a Tenax or a trilayer sorbent were tested because of their selective affinity for volatiles. Then, innovative variations of these methods were developed to combine their respective advantages in a one-step extraction: Static headspace with multiple SPME fibers (S-HS-MultiSPME), Dynamic headspace with multiple SPME fibers (D-HS-MultiSPME) and dynamic headspace with multiple SPME fibers and Tenax (D-HS-MultiSPME/Tenax). The extracts were analyzed by gas chromatography coupled with mass spectrometric and flame ionization detection. The relative performances of these methods were compared based on qualitative and semi-quantitative analyses of the chromatograms. The D-HS technique showed good sensitivity for most compounds, whereas HS-SPME favored the extraction of acids. The D-HS-MultiSPME/Tenax identified more than 60 compounds from human milk (some for the first time) and evidence of individual singularities. This method that can be applied to volatilome analysis of any biological fluid should further our understanding of human milk odor.

Male mice and cows perceive human emotional chemosignals: a preliminary study.

Olfactory cues of individuals of the same species or from different species may induce changes in behaviors and physiological reactions in mammals. However, there are few studies on the influence of human odor on animal behavior and welfare, especially those of rodents and farm animals. The present study aimed to investigate whether the odor of a stressed human (in sweat) would modify the behavior of mice and cows. We hypothesized that laboratory and farm animals can perceive human emotions though olfactory cues and that human emotional chemosignals can modify their behavioral reactions and welfare. Two odors of human axillary sweat were collected from engineering students (n = 25, 14 females and 11 males; 21.1 ± 0.7 years old, range: 19-23 years old): a "stress" odor collected after an exam and a "non-stress" odor collected after a standard class. Two experiments were then conducted to test the discrimination of these two odors by male mice (n = 20) under standard conditions and by cows (n = 10) under farm conditions. During the experiments, the behavioral responses of the animals to both odors (through a dispenser for the mice and a bucket for the cows) were observed. The mice produced significantly (p = 0.004) more fecal pellets with the stress odor dispenser than with the non-stress-odor dispenser. The cows spent significantly (p = 0.04) more time smelling the non-stress-odor bucket than control. For both species, the other behaviors observed did not differ significantly between the odors. Mice and cows seemed to perceive and react to stressful human chemosignals. Mice showed physiological reactions that indicated stress in response to the stress odor of humans, while cows showed preference reactions in response to the non-stress odor of humans. This preliminary study showed that laboratory and farm animals, such as male mice and cows, seemed to discriminate certain odors emitted by humans that were likely related to different emotions. Animals may recognize stressful human chemosignals, associate these signals with negative husbandry practices or human-animal relationships, and consequently modify their behavior.

Newborn crawling and rooting in response to maternal breast odor.

A growing literature shows that perception and action are already tightly coupled in the newborn. The current study aimed to examine the nature of the coupling between olfactory stimuli from the mother and the newborn's crawling and rooting (exploratory movements of the head). To examine the coupling, the crawling and rooting behavior of 28 2-day-old newborns were studied while they were supported prone on a mobility device shaped like a mini skateboard, the Crawliskate®, their head positioned directly on top of a pad infused with either their mother's breast odor (Maternal) or the odor of water (Control). Video and 3D kinematic analyses of the number and types of limb movements and quantification of displacement across the surface revealed that newborns are significantly more efficient crawlers when they smell the maternal odor, moving greater distances although performing fewer locomotor movements. In addition, the newborns made significantly more head rooting movements in the presence of the maternal odor. These findings suggest that the circuitry underlying quadrupedal locomotion and exploratory movements of the head is already adaptable to olfactory information via higher brain processing. Moreover, the coupling between olfaction and the two action systems, locomotion and rooting, is already differentiated. As crawling enables the newborn to move toward the mother's breast immediately after birth and facilitates mother-infant interaction, the results of this study highlight the potential value of using maternal odors to stimulate mobility in infants at risk of motor delay and/or deprived of this odor when born prematurely.

Human neonates prefer colostrum to mature milk: Evidence for an olfactory bias toward the "initial milk"?

OBJECTIVES: Colostrum is the initial milk secretion which ingestion by neonates warrants their adaptive start in life. Colostrum is accordingly expected to be attractive to newborns. The present study aims to assess whether colostrum is olfactorily attractive for 2-day-old newborns when presented against mature milk or a control. METHODS: The head-orientation of waking newborns was videotaped in three experiments pairing the odors of: (a) colostrum (sampled on postpartum day 2, not from own mother) and mature milk (sampled on average on postpartum day 32, not from own mother) (n tested newborns = 15); (b) Colostrum and control (water; n = 9); and (c) Mature milk and control (n = 13). RESULTS: When facing the odors of colostrum and mature milk, the infants turned their nose significantly longer toward former (32.8 vs 17.7% of a 120-s test). When exposed to colostrum against the control, they responded in favor of colostrum (32.9 vs 16.6%). Finally, when the odor of mature milk was presented against the control, their response appeared undifferentiated (26.7 vs 28.6%). CONCLUSIONS: These results indicate that human newborns can olfactorily differentiate conspecific lacteal fluids sampled at different lactation stages. They prefer the odor of the mammary secretion - colostrum - collected at the lactation stage that best matches the postpartum age of their own mother. These results are discussed in the context of the earliest mother-infant chemo-communication. Coinciding maternal emission and offspring reception of chemosignals conveyed in colostrum may be part of the sensory precursors of attunement between mothers and infants.

The role of papillary skin glands in guiding mouse pups to the nipple.

The nipple odor of lactating mice (Mus musculus) plays a crucial role in attracting newborn pups and motivating them to suck milk. The characteristic odor of a lactating murine nipple is assumed to be a mixture of multiple odorous substrates, that is, milk, dam's and pups' saliva, skin glands' secretions, and amniotic fluid. The present study aimed to characterize the behavioral activity of the original odor mixture that develops over the nipples in the first 2 days postpartum. We extracted this odor mixture in water and evaluated its attractive and appetitive potencies using two behavioral assays (viz., relative attraction and oral activation assays). It resulted that the so-called nipple wash was as appetitive as fresh milk, and even more attractive than it. The behavioral potency of the nipples was shown to be specific to lactating nipples (relative to nulliparous nipples) and to be preserved for 2 weeks when stored at -80°C. Finally, we perfected a nipple deodorization procedure by inactivating the nipples' behavioral potency. We observed that such altered appetitive potency was fully restored 30 min after its washing, but without any maternal self-licking and pups' sucking, indicating that the secretions of the nipple skin glands' were sufficient to explain the success of neonatal guidance to the nipple.

Maternal odor shapes rapid face categorization in the infant brain.

To successfully interact with a rich and ambiguous visual environment, the human brain learns to differentiate visual stimuli and to produce the same response to subsets of these stimuli despite their physical difference. Although this visual categorization function is traditionally investigated from a unisensory perspective, its early development is inherently constrained by multisensory inputs. In particular, an early-maturing sensory system such as olfaction is ideally suited to support the immature visual system in infancy by providing stability and familiarity to a rapidly changing visual environment. Here, we test the hypothesis that rapid visual categorization of salient visual signals for the young infant brain, human faces, is shaped by another highly relevant human-related input from the olfactory system, the mother's body odor. We observe that a right-hemispheric neural signature of single-glance face categorization from natural images is significantly enhanced in the maternal versus a control odor context in individual 4-month-old infant brains. A lack of difference between odor conditions for the common brain response elicited by both face and non-face images rules out a mere enhancement of arousal or visual attention in the maternal odor context. These observations show that face-selective neural activity in infancy is mediated by the presence of a (maternal) body odor, providing strong support for multisensory inputs driving category acquisition in the developing human brain and having important implications for our understanding of human perceptual development.

Does any mother's body odor stimulate interest in mother's face in 4-month-old infants?

Little is known about the effects of olfaction on visual processing during infancy. We investigated whether and how an infant's own mother's body odor or another mother's body odor affects 4-month-old infants' looking at their mother's face when it is paired with a stranger's face. In Experiment 1, infants were exposed to their mother's body odor or to a control odor, while in Experiment 2, infants were exposed to a stranger mother's body odor while their visual preferences were recorded. Results revealed that infants looked more at the stranger's female face in presence of the control odor but that they looked more at their mother's face in the context of any mother's body odors. This effect was due to a reduction of looking at the stranger's face. These findings suggest that infants react similarly to the body odor of any mother and add to the growing body of evidence indicating that olfactory stimulation represents a pervasive aspect of infant multisensory perception.

Weanling Infants Prefer the Odors of Green Vegetables, Cheese, and Fish When Their Mothers Consumed These Foods During Pregnancy and/or Lactation.

Infants' olfactory experience begins before birth and extends after birth through milk and complementary foods. Until now, studies on the effects of chemosensory experience in utero and/or through human milk focused on experimentally controlled exposure to only 1 target food bearing a specific odor quality and administered in sizeable amounts. This study aimed to assess whether early olfactory experience effect was measurable in "everyday conditions" of maternal food intake during pregnancy and lactation, and of infant intake at weaning, leading to expose the infant to corresponding odors as fetus, neonate, and infant up to 8 and 12 months of age. Infants' early food exposures were assessed by asking mothers to fill out diaries about their food consumption during pregnancy and breastfeeding, and about their infant's consumption during complementary feeding. To test odor liking, odorants representing a priori pleasant and unpleasant food odors, as well as odorless stimuli, were presented. The infant's exploratory behavior toward odorized bottles and nonodorized control bottles was measured in terms of mouthing duration, which is thought to reflect attraction and/or appetence. At age 8 months only, positive correlations were found between liking of some unpleasant odors and early exposure to these odors through mother's diet. No correlations were found between infants' liking of the pleasant odors and early exposures to the foods bearing these odors. This study highlights that early exposure to unpleasant food odors may increase subsequent liking (or reduce subsequent dislike) of these food odors at least until the age of 8 months.

Chemical Profiles of Integumentary and Glandular Substrates in Australian Sea Lion Pups (Neophoca cinerea).

Recognition of individuals or classes of individuals plays an important role in the communication systems of many mammals. The ability of otariid (i.e., fur seal and sea lion) females to locate and identify their offspring in colonies after returning from regular foraging trips is essential to successful pup rearing. It has been shown that olfaction is used to confirm the identity of the pup by the mother when they reunite, yet the processes by which this chemical recognition occurs remain unclear. Using gas chromatography-mass spectrometry, we examined chemical profiles of integumentary and glandular secretions/excretions from pre- and post-molt Australian sea lion pups (Neophoca cinerea) and compared fur and swab samples to assess data collection methods. Multivariate statistics were applied to assess differences in chemical composition between body regions and sexes. We found differences among secretions from various body regions, driven by the distinctiveness of the oral odor mixture. The fine-scale trends in pre- and post-molt pups seem to differ due to changes in the behavior of pups and consequent decrease in the transfer of compounds among adjacent body regions in older pups. Volatile compounds from exocrine substrates were not distinct for different sexes. We also show that swab samples provide better data for exploring social olfaction than fur samples for this species. Obtaining fundamental chemical information, in this case chemical profiles of animals, and discerning differences in chemical composition is an important step toward fully exploring the intricacies of mother-offspring olfactory recognition and its underlying processes.

Watching happy faces potentiates incentive salience but not hedonic reactions to palatable food cues in overweight/obese adults.

'Wanting' and 'liking' are mediated by distinct brain reward systems but their dissociation in human appetite and overeating remains debated. Further, the influence of socioemotional cues on food reward is little explored. We examined these issues in overweight/obese (OW/OB) and normal-weight (NW) participants who watched food images varying in palatability in the same time as videoclips of avatars looking at the food images while displaying facial expressions (happy, disgust or neutral) with their gaze directed only toward the food or consecutively toward the food and participants. We measured heart rate (HR) deceleration as an index of attentional/incentive salience, facial EMG activity as an index of hedonic or disgust reactions, and self-report of wanting and liking. OW/OB participants exhibited a larger HR deceleration to palatable food pictures than NW participants suggesting that they attributed greater incentive salience to food cues. However, in contrast to NW participants, they did not display increased hedonic facial reactions to the liked food cues. Subjective ratings of wanting and liking did not differentiate the two groups. Further, OW/OB participants had more pronounced HR deceleration than NW participants to palatable food cues when they watched avatars' happy faces gazing at the food. In line with the "incentive-sensitization" hypothesis, our data suggest that incentive salience attribution and not hedonic reactivity is increased in OW/OB individuals and that happy faces, as social reward cues, potentiate implicit wanting in OW/OB people.

Tuning functions for automatic detection of brief changes of facial expression in the human brain.

Efficient decoding of even brief and slight intensity facial expression changes is important for social interactions. However, robust evidence for the human brain ability to automatically detect brief and subtle changes of facial expression remains limited. Here we built on a recently developed paradigm in human electrophysiology with full-blown expressions (Dzhelyova et al., 2017), to isolate and quantify a neural marker for the detection of brief and subtle changes of facial expression. Scalp electroencephalogram (EEG) was recorded from 18 participants during stimulation of a neutral face changing randomly in size at a rapid rate of 6 Hz. Brief changes of expression appeared every five stimulation cycle (i.e., at 1.2 Hz) and expression intensity increased parametrically every 20 s in 20% steps during sweep sequences of 100 s. A significant 1.2 Hz response emerged in the EEG spectrum already at 40% of facial expression-change intensity for most of the 5 emotions tested (anger, disgust, fear, happiness, or sadness in different sequences), and increased with intensity steps, predominantly over right occipito-temporal regions. Given the high signal-to-noise ratio of the approach, thresholds for automatic detection of brief changes of facial expression could be determined for every single individual brain. A time-domain analysis revealed three components, the two first increasing linearly with increasing intensity as early as 100 ms after a change of expression, suggesting gradual low-level image-change detection prior to visual coding of facial movements. In contrast, the third component showed abrupt sensitivity to increasing expression intensity beyond 300 ms post expression-change, suggesting categorical emotion perception. Overall, this characterization of the detection of subtle changes of facial expression and its temporal dynamics open promising tracks for precise assessment of social perception ability during development and in clinical populations.

Seeing odors in color: Cross-modal associations in children and adults from two cultural environments.

We investigated the occurrence and underlying processes of odor-color associations in French and American 6- to 10-year-old children (n = 386) and adults (n = 137). Nine odorants were chosen according to their familiarity to either cultural group. Participants matched each odor with a color, gave hedonic and familiarity judgments, and identified each odor. By 6 years of age, children displayed culture-specific odor-color associations, but age differences were noted in the type of associations. Children and adults in both cultural groups shared common associations and formed associations that were unique to their environment, underscoring the importance of exposure learning in odor-color associations.

Mimicking emotions: how 3-12-month-old infants use the facial expressions and eyes of a model.

While there is an extensive literature on the tendency to mimic emotional expressions in adults, it is unclear how this skill emerges and develops over time. Specifically, it is unclear whether infants mimic discrete emotion-related facial actions, whether their facial displays are moderated by contextual cues and whether infants' emotional mimicry is constrained by developmental changes in the ability to discriminate emotions. We therefore investigate these questions using Baby-FACS to code infants' facial displays and eye-movement tracking to examine infants' looking times at facial expressions. Three-, 7-, and 12-month-old participants were exposed to dynamic facial expressions (joy, anger, fear, disgust, sadness) of a virtual model which either looked at the infant or had an averted gaze. Infants did not match emotion-specific facial actions shown by the model, but they produced valence-congruent facial responses to the distinct expressions. Furthermore, only the 7- and 12-month-olds displayed negative responses to the model's negative expressions and they looked more at areas of the face recruiting facial actions involved in specific expressions. Our results suggest that valence-congruent expressions emerge in infancy during a period where the decoding of facial expressions becomes increasingly sensitive to the social signal value of emotions.

Responses of Human Neonates to Highly Diluted Odorants from Sweat.

Conjugated forms of odorants contributing to sweat odor occur not only in human sweat but also in amniotic fluid, colostrum, and milk. However, it is unclear whether the released odorants are detected and hedonically discriminated by human newborns. To investigate this issue, we administered highly diluted solutions of (R)/(S)-3-methyl-3-sulfanylhexan-1-ol (MSH), (R)/(S)-3-sulfanylhexan-1-ol (SH), (E)/(Z)-3-methylhex-2-enoic acid (3M2H), and (R)/(S)-3-hydroxy-3-methylhexanoic acid (HMHA) to 3-d-old infants while their respiratory rate and oro-facial movements were recorded. Adult sensitivity to these odorants was assessed via triangle tests. Whereas no neonatal stimulus-specific response was found for respiratory rate, oro-facial reactivity indicated orthonasal detection of MSH and SH by male neonates, and of HMHA by the whole group of neonates. Dependent on the dilution of odorants, newborns evinced neutral responses or longer negative oro-facial expressions compared with the reference stimuli. Finally, newborns appeared to be more sensitive to the target odorants than did adults.

Affective matching of odors and facial expressions in infants: shifting patterns between 3 and 7 months.

Recognition of emotional facial expressions is a crucial skill for adaptive behavior. Past research suggests that at 5 to 7 months of age, infants look longer to an unfamiliar dynamic angry/happy face which emotionally matches a vocal expression. This suggests that they can match stimulations of distinct modalities on their emotional content. In the present study, olfaction-vision matching abilities were assessed across different age groups (3, 5 and 7 months) using dynamic expressive faces (happy vs. disgusted) and distinct hedonic odor contexts (pleasant, unpleasant and control) in a visual-preference paradigm. At all ages the infants were biased toward the disgust faces. This visual bias reversed into a bias for smiling faces in the context of the pleasant odor context in the 3-month-old infants. In infants aged 5 and 7 months, no effect of the odor context appeared in the present conditions. This study highlights the role of the olfactory context in the modulation of visual behavior toward expressive faces in infants. The influence of olfaction took the form of a contingency effect in 3-month-old infants, but later evolved to vanish or to take another form that could not be evidenced in the present study.

Emotional expressiveness of 5-6 month-old infants born very premature versus full-term at initial exposure to weaning foods.

Facial expressions of 5-6 month-old infants born preterm and at term were compared while tasting for the first time solid foods (two fruit and two vegetable purées) given by the mother. Videotapes of facial reactions to these foods were objectively coded during the first six successive spoons of each test food using Baby FACS and subjectively rated by naïve judges. Infant temperament was also assessed by the parents using the Infant Behaviour Questionnaire. Contrary to our expectations, infants born preterm expressed fewer negative emotions than infants born full-term. Naïve judges rated infants born preterm as displaying more liking than their full-term counterparts when tasting the novel foods. The analysis of facial expressions during the six spoonfuls of four successive meals (at 1-week intervals) suggested a familiarization effect with the frequency of negative expressions decreasing after tasting the second spoon, regardless of infant age, type of food and order of presentation. Finally, positive and negative dimensions of temperament reported by the parents were related with objective and subjective coding of affective reactions toward foods in infants born preterm or full-term. Our research indicates that premature infants are more accepting of novel foods than term infants and this could be used for supporting the development of healthy eating patterns in premature infants. Further research is needed to clarify whether reduced negativity by infants born prematurely to the exposure to novel solid foods reflects a reduction of an adaptive avoidant behaviour during the introduction of novel foods.

Responsiveness of human neonates to the odor of 5α-androst-16-en-3-one: a behavioral paradox?

The odorous steroid 5α-androst-16-en-3-one (AND) occurs in numerous biological fluids in mammals, including man, where it is believed to play a chemocommunicative role. As AND was recently detected in milk and amniotic fluid, sensitivity and hedonic responses to this substance were assessed in human neonates. To this aim, respiration and facial expressions were recorded in 3-day-old newborns in response to aqueous solutions of AND, ranging from 500ng/mL to 0.5 fg/mL. Although analyses of respiratory rate did not lead to clear-cut results, the newborns changed their facial expressions at concentrations not detected by adults in a triangle test. Newborns displayed negative facial actions of longer duration to AND relative to an odorless control. Thus, AND may be considered to be offensive to newborns, which is a counterintuitive outcome as they are exposed to this compound in the womb (and it should therefore be familiar), in milk, and on the mother's skin surface (and it should therefore be conditioned as positive). Multiple reasons for this perceptual-behavioral paradox are discussed.