Direct Behavioral and Neurophysiological Evidence for Retronasal Olfaction in Mice

The neuroscience of flavor perception is hence becoming increasingly important to understand food flavor perception that guides food selection, ingestion and appreciation. We recently provided evidence that rats can use the retronasal mode of olfaction, an essential element of human flavor perception. We showed that in rats, like humans, odors can acquire a taste. We and others also defined how the input of the olfactory bulb (OB) -not functionally imageable in humans- codes retronasal smell in anesthetized rat. The powerful awake transgenic mouse, however, would be a valuable additional model in the study of flavor neuroscience. We used a go/no-go behavioral task to test the mouse''s ability to detect and discriminate the retronasal odor amyl acetate. In this paradigm a tasteless aqueous odor solution was licked by water-restricted head-fixed mice from a lick spout. Orthonasal contamination was avoided. The retronasal odor was successfully discriminated by mice against pure distilled water in a concentration-dependent manner. Bulbectomy removed the mice''s ability to discriminate the retronasal odor but not tastants. The OB showed robust optical calcium responses to retronasal odorants in these awake mice. These results suggest that mice, like rats, are capable of smelling retronasally. This direct neuro-behavioral evidence establishes the mouse as a useful additional animal model for flavor research.     

The role of congruency in retronasal odor referral to the mouth

Referral of retronasal odors to the mouth is a fundamental phenomenon of flavor perception. A previous study from this laboratory provided evidence that, contrary to prior speculation, taste rather than touch was the primary factor in retronasal odor referral. The present study further investigated this question by studying the role of congruency between taste and odor on retronasal odor referral under conditions that mimicked natural food consumption. Subjects performed odor localization tasks after sampling gelatin stimuli that contained various congruent and incongruent tastes-odor combinations. The results showed that when a congruent taste was added, referral to the oral cavity and tongue were significantly enhanced. In addition, the data also indicate that the degree of congruency between taste and odor may modulate the degree of odor referral to the mouth. These findings suggest that odor referral is maximized when congruent flavor dimensions are combined to trigger perceptual "flavor objects" that represent known or potential foods. The results are discussed in terms of the factors that play a role in the retronasal odor referral as well as the potential neural mechanisms that may underlie it.     

fMRI activation in response to odorants orally delivered in aqueous solutions

During food intake flavor perception results from simultaneous stimulation of the gustatory, olfactory and trigeminal systems. Olfactory stimulation occurs mainly through the retronasal pathway and the resulting perception is often interpreted as a taste perception, thus leading to the well-known sensory confusion between taste and olfaction. The present experiment was designed to study, with functional magnetic resonance imaging (fMRI), the cortical representation of olfactory perception in humans in response to retronasal stimulation by odorants delivered in aqueous solution. Psychophysical evaluation confirmed that the stimuli acted as pure olfactory stimuli through the retronasal pathway and did not present any taste component. Results showed activation in all brain regions previously described with neuroimaging techniques using olfactory stimulation with an odorized air flow. Piriform and orbitofrontal cortex were found activated as well as the hippocampal region, the amygdala, the insular lobe, the cingulate gyrus and the cerebellum. These results demonstrate the feasibility of efficiently stimulating the olfactory system in an fMRI scanner through the retronasal pathway with liquids delivered to the oral cavity. The presentation of olfactory stimuli in liquids to the mouth is a realistic model for the study of food-related flavor perception. This stimulation protocol furthermore allows presenting taste and olfactory stimuli separately or combined, thus allowing for direct comparisons between single modality representation, taste or olfaction, and representation of multi-modality mixtures.     

Enhancement of retronasal odors by taste

Psychophysical studies of interactions between retronasal olfaction and taste have focused most often on the enhancement of tastes by odors, which has been attributed primarily to a response bias (i.e., halo dumping). Based upon preliminary evidence that retronasal odors could also be enhanced by taste, the present study measured both forms of enhancement using appropriate response categories. In the first experiment, subjects rated taste ("sweet," "sour," "salty," and "bitter") and odor ("other") intensity for aqueous samples of 3 tastants (sucrose, NaCl, and citric acid) and 3 odorants (vanillin, citral, and furaneol), both alone and in taste-odor mixtures. The results showed that sucrose, but not the other taste stimuli, significantly increased the perceived intensity of all 3 odors. Enhancement of tastes by odors was inconsistent and generally weaker than enhancement of odors by sucrose. A second experiment used a flavored beverage and a custard dessert to test whether the findings from the first experiment would hold for the perception of actual foods. Adding sucrose significantly enhanced the intensity of "cherry" and "vanilla" flavors, whereas adding vanillin did not significantly enhance the intensity of sweetness. It is proposed that enhancement of retronasal odors by a sweet stimulus results from an adaptive sensory mechanism that serves to increase the salience of the flavor of nutritive foods.     

Measurement of sensitivity to olfactory flavor: application in a study of aging and dentures

Olfaction involves a dual sensory process for perceiving odors orthonasally (through the nostrils) and retronasally (through the mouth). This investigation entailed developing a measure of sensitivity to an odor delivered in an orally sampled food (orange flavoring in a sucrose-sweetened gelatin) and examining sensitivity in the elderly. In experiment 1, olfactory flavor sensitivity was 49 times lower in elderly (n = 21) than in young (n = 28) subjects. In experiment 2, with 73 elderly women, higher olfactory flavor sensitivity correlated significantly with higher orthonasal perception (Connecticut Chemosensory Clinical Research Center test). Some women, however, exhibited low olfactory flavor sensitivity despite high orthonasal perception; none had high olfactory flavor sensitivity and low orthonasal perception. Those who wore complete or palatal covering dentures had lower olfactory flavor sensitivity than those who were dentate or wore dentures that did not cover the palate. Through multiple regression analysis, orthonasal perception and denture status were found to be independent contributors to predicting olfactory flavor sensitivity. In summary, elderly subjects showed depressed olfactory flavor sensitivity (i.e. retronasal sensitivity) that related to poor orthonasal olfactory perception and denture characteristic. Thus, while good orthonasal olfaction may be necessary for good olfactory flavor sensitivity, it is not sufficient. Other factors, some associated with oral conditions, may impede release and retronasal transport of odors from the mouth to the olfactory receptors.     

Direct behavioral evidence for retronasal olfaction in rats

The neuroscience of flavor perception is becoming increasingly important to understand abnormal feeding behaviors and associated chronic diseases such as obesity. Yet, flavor research has mainly depended on human subjects due to the lack of an animal model. A crucial step towards establishing an animal model of flavor research is to determine whether the animal uses the retronasal mode of olfaction, an essential element of flavor perception. We designed a go- no go behavioral task to test the rat's ability to detect and discriminate retronasal odorants. In this paradigm, tasteless aqueous solutions of odorants were licked by water-restricted head-fixed rats from a lick spout. Orthonasal contamination was avoided by employing a combination of a vacuum around the lick-spout and blowing clean air toward the nose. Flow models support the effectiveness of both approaches. The licked odorants were successfully discriminated by rats. Moreover, the tasteless odorant amyl acetate was reliably discriminated against pure distilled water in a concentration-dependent manner. The results from this retronasal odor discrimination task suggest that rats are capable of smelling retronasally. This direct behavioral evidence establishes the rat as a useful animal model for flavor research.     

Food perception with age and its relationship to pleasantness

Differences between elderly subjects (n = 46, 61-86 years) and young subjects (n = 36, 18-25 years) in food perception and food liking were investigated. Intensity and liking ratings were assessed for custard dessert, in which flavor enrichment, textural change, and irritant addition were incorporated as strategies to compensate for sensory losses with increasing age. The sensory acuity (taste, olfaction, irritation, chewing efficiency) of both young and elderly subjects was measured with the help of different sensitivity tests. The elderly perceived the custards differently from the young, mainly as less intense in flavor (cherry/vanilla) and less intense in creaminess/swallowing effort. Several of the observed interaction effects were different for the elderly and the young. The majority of these differences manifested as lower intensity slopes for the elderly. Losses in sensitivity to taste and to olfactory and trigeminal stimuli as well as a reduced chewing efficiency were observed on average for the elderly compared with the young. Furthermore, subgroups of the elderly were observed in which the compensatory strategies flavor enrichment, textural change, and irritant addition led to an increase in food liking. However, these subgroups did not differ in their sensory acuity. The present study does not support the assumption that age-associated changes in food perception-caused by losses in sensory acuity-inevitably reduce the food liking of the elderly.     

Identifying Breeding Priorities for Blueberry Flavor Using Biochemical,Sensory, and Genotype by Environment Analyses

Breeding for a subjective goal such as flavor is challenging, as many blueberry cultivars are grown worldwide, and identifying breeding targets relating to blueberry flavor biochemistry that have a high degree of genetic control and low environmental variability are priorities. A variety of biochemical compounds and physical characters induce the sensory responses of taste, olfaction, and somatosensation, all of which interact to create what is perceived flavor. The goal of this study was to identify the flavor compounds with a larger genetic versus environmental component regulating their expression over an array of cultivars, locations, and years. Over the course of three years, consumer panelists rated overall liking, texture, sweetness, sourness, and flavor intensity of 19 southern highbush blueberry (Vaccinium corymbosum hybrids) genotypes in 30 sensory panels. Significant positive correlations to overall liking of blueberry fruit (P<0.001) were found with sweetness (R² = 0.70), texture (R² = 0.68), and flavor (R² = 0.63). Sourness had a significantly negative relationship with overall liking (R² = 0.55). The relationship between flavor and texture liking was also linear (R² = 0.73, P<0.0001) demonstrating interaction between olfaction and somatosensation. Partial least squares analysis was used to identify sugars, acids, and volatile compounds contributing to liking and sensory intensities, and revealed strong effects of fructose, pH, and several volatile compounds upon all sensory parameters measured. To assess the feasibility of breeding for flavor components, a three year study was conducted to compare genetic and environmental influences on flavor biochemistry. Panelists could discern genotypic variation in blueberry sensory components, and many of the compounds affecting consumer favor of blueberries, such as fructose, pH, β-caryophyllene oxide and 2-heptanone, were sufficiently genetically controlled that allocating resources for their breeding is worthwhile.     

Taste-smell interactions are tastant and odorant dependent

Four experiments were conducted to assess the nature of taste–smellinteractions. In the first experiment, the ability of strawberryodor to modify the sweetness of sucrose was investigated. Thiswas accomplished by having subjects rate the sweetness of whipped-creamstimuli with and without strawberry odor over time. The stimuliwere swallowed to augment retronasal stimulation of the olfactorysystem. It was found that strawberry odor tended to enhancethe maximal sweetness and total rating time of the stimuli.In the second experiment, it was found that peanut butter odordid not enhance sweetness, thus demonstrating that an odor'sability to enhance sweetness is odor-dependent, In the thirdexperiment, it was demonstrated that strawberry odor did notenhance the saltiness of sodium chloride indicating that anodor's ability to enhance taste is tastant-dependent. In thefourth experiment, it was shown that 85% of the strawberry odorant'sability to enhance sweetness was eliminated by pinching thenostrils. This suggests that the influence of the strawberryodorant on sweetness was olfactory rather than gustatory. Itwas concluded that an odor's influence on taste is both odorantand tastant dependent.     

SENSORY PROPERTIES AFFECTING ACCEPTABILITY OF "BRESAOLA" FROM PODOLIAN YOUNG BULLS

Three muscles ( Semimembranosus, Quadriceps femoris, Adductor ) from Podolian young bulls (aged 18 months) were processed into bresaolas, which were compared with commercial products, in terms of sensory profile and preference expressed by consumers. Slope analysis was conducted to study the preference drivers. The preference of homogeneous groups of consumers and the relationships with sensory properties of bresaola were investigated using partial least squares (PLS) regression analysis. The products differed for several sensory properties, such as sweetness, flavor intensity ( P <  0.001), saltiness ( P <  0.01), peppered, seasoned and tenderness ( P <  0.05). The slope analysis showed that taste/flavor ( k =  0.86) and appearance ( k =  0.76) play an important role in orienting consumer preference, while texture ( k =  0.54) is less important. No general trend was found for product liking. Conversely, PLS regression clustered the consumers in homogeneous groups: one preferred the commercial products characterized by sensory properties such as peppered, saltiness, seasoned and marbling, another one was more oriented toward properties such as sweetness, odor and chewiness, which characterized Podolian bresaolas.

PRACTICAL APPLICATION

Partial least squares regression analysis clustered the consumers in homogeneous groups according to their liking: those preferring peppered, salty, seasoned and marbling bresaolas versus those oriented toward sweet and odorous products. In addition, the relationship between overall liking and attribute liking (appearance, taste/flavor and texture) was analyzed by linear regression analysis. This may be helpful in order to establish meaningful criteria for grading bresaolas and identify the particular attributes that move its acceptance (i.e., taste/flavor and appearance rather than texture).     

Observation of the swallowing process by application of videofluoroscopy and real-time magnetic resonance imaging-consequences for retronasal aroma stimulation.

The process of eating and drinking was observed in vivo by application of videofluoroscopy, a dynamic X-ray technique, as well as real-time magnetic resonance imaging. The study was aimed at elucidating the timing and performance of the physiological organs involved in mastication and swallowing, mainly the tongue, the pharynx and the soft palate (velum palatinum). It was shown for the first time that effective physiological barriers do exist during food consumption that are capable of retaining volatiles such as helium within the oral cavity. These barriers allow the access of odorants to the nasal cavity only at certain times during the eating process. Their effectiveness is related to the texture of the food as well as the amount of food material present in the oral cavity and, thereby, directly influences retronasal aroma perception.     

THE EFFECTS OF SUGARS AND ACIDS ON CONSUMER ACCEPTABILITY OF KIWIFRUIT

A kiwifruit (Actinidia deliciosa (A Chev) Liang et Ferguson cv Hayward) pulp was made using fruit harvested immature to minimize the presence of aroma volatiles. This pulp was modified by incorporating sugars and acids to simulate kiwifruit with a range of Brix and acid levels. Using sensory evaluation, flavor acceptability was found to increase with increasing Brix. As expected, perceived " sweetness intensity" increased with increasing Brix. Acidity did not influence flavor acceptability. At high Brix, sugars were able to suppress the effects of variations in acidity. Changes in Brix did not influence " flavor intensity", suggesting that aroma volatiles may be important contributors to kiwifruit flavor intensity.     

Potential mechanisms of retronasal odor referral to the mouth

The current study took a first step toward elucidating the sensory input that drives retronasal odor referral to the mouth. In 2 experiments, subjects performed odor localization tasks under various oral-nasal stimulation conditions that allowed us to assess the effects of direction of airflow, taste, and tactile stimulation on retronasal odor referral. Subjects reported the locations of perceived odors when food odorants were inhaled through the mouth alone or in the presence of water or various tastants in the mouth. The results indicated that when perceived alone, vanilla and soy sauce odor were localized 54.7%: 26.4%: 18.9% and 60.0%: 21.7%: 18.3% in the nose, oral cavity, and on the tongue, respectively. The localization of odors alone was not significantly different from when water was presented simultaneously in the mouth, indicating that tactile stimulation itself is not sufficient to enhance odor referral. However, the presence of sucrose, but not other tastes, significantly increased localization of vanilla to the tongue. Likewise, only NaCl significantly augmented referral of soy sauce odor to the tongue. These data indicate that referral of retronasal odors to the mouth can occur in the absence of a either taste or touch but that referral to the tongue depends strongly on the presence of a congruent taste.     

COMPARISON OF ATTRIBUTE LIKING AND JAR SCALES TO EVALUATE THE ADEQUACY OF SENSORY ATTRIBUTES OF MILK DESSERTS

Just-about-right (JAR) scales and attribute liking questions are usually used to study consumer perception of the sensory characteristics of food products. The aim of the present work was to compare the performance of attribute liking and JAR scales to evaluate consumers' perceived adequacy of flavor and texture of milk puddings. Two groups of consumers were asked to evaluate eight milk desserts using (1) overall liking followed by attribute liking for texture and flavor and (2) overall liking followed by JAR scales for thickness, creaminess, sweetness and vanilla flavor. Overall liking scores were significantly different when JAR scales or attribute liking questions were considered. Texture, flavor and overall liking scores were highly correlated to each other, providing the same information. JAR scales correlated better with the intensity of sensory attributes evaluated by a trained sensory panel, being JAR percentages a reliable tool to study the adequacy of sensory attributes.

PRACTICAL APPLICATIONS

Results from the present work showed that consumers might not be able to independently evaluate their liking of different sensory attributes of a product. For this reason, the use of attribute liking questions for studying the adequacy of sensory attributes in complex products would not be recommended. JAR scales were better indicators of the adequacy of sensory attributes. Consumers were able to independently evaluate texture and flavor attributes using JAR scales. However, the influence of JAR scales on overall liking scores should be taken into account when including these scales on consumer studies.     

Revisiting sugar-fat mixtures: sweetness and creaminess vary with phenotypic markers of oral sensation

Genetic variation in oral sensation presumably influences ingestive behaviors through sensations arising from foods and beverages. Here, we investigated the influence of taste phenotype [6-n-propylthiouracil (PROP) bitterness, fungiform papillae (FP) density] on sweet and creamy sensations from sugar/fat mixtures. Seventy-nine subjects (43 males) reported the sweetness and creaminess of water or milk (skim, whole, heavy cream) varying in sucrose (0-20% w/v) on the general Labeled Magnitude Scale. Sweetness grew with sucrose concentration and when shifting from water to milk mixtures--the growth was greatest for those tasting PROP as most bitter. At higher sucrose levels, increasing fat blunted the PROP-sweet relationship, whereas at lower levels, the relationship was effectively eliminated. Perceived sweetness of the mixture exceeded that predicted from the sum of components at low sucrose concentrations (especially for those tasting PROP most bitter) but fell below predicted at high concentrations, irrespective of fat level. Creaminess increased greatly with fat level and somewhat with sucrose. Those tasting PROP most bitter perceived greater creaminess in the heavy cream across all sucrose levels. Perceived creaminess was somewhat lower than predicted, irrespective of PROP bitterness. The FP density generally showed similar effects as PROP on sweetness and creaminess, (but to a lesser degree) and revealed potential taste-somatosensory interactions in weakly sweet stimuli. These data support that taste phenotype affects the nature of enhancement or suppression of sweetness and creaminess in liquid fat/sugar mixtures. Taste phenotype effects on sweetness and creaminess likely involve differential taste, retronasal olfactory, and somatosensory contributions to these perceptual experiences.     

Role of hydroxycinnamic acids in food flavor: a brief overview

The influence of hydroxycinnamic acids (HCAs) on food flavor is reviewed. In coffee, whole-grain foods and related model systems, the HCAs have been reported to contribute to the flavor profile by multiple mechanisms, such as to impart taste attributes, to generate aroma and taste-active compounds by phenolic degradation, as well as to alter the mechanisms of the Maillard reaction and related flavor development. Consequently the role of HCAs on the flavor properties of food products is complex, multifaceted and can be related to the chemistry and fate of HCAs during thermal processing.     

Sensory evaluation of fruit quality and nutritional composition of 20 breadfruit (Artocarpus, Moraceae) cultivars

Twenty breadfruit cultivars growing in afield genebank at Kahanu Garden, National Tropical Botanical Garden, Hana, Maui, Hawaii, were evaluated for sensory attributes and nutrient composition. A taste panel scored eight flavor/aroma attributes, five textural attributes, and color. There were significant differences (P ≤ 0.01) in aroma, visual texture, flavor intensity, sweetness, starchiness, moistness, stringiness, firmness, and color. The greatest differences were in color and texture. Nutrient analyses showed significant differences (P ≤ 0.05) for energy, carbohydrates, ash, crude protein, potassium, magnesium, sodium, iron, copper, and zinc. Considering the versatility of breadfruit as a food, its ease of production, and its nutritional value, the numerous good quality flavorful cultivars available should be more widely grown for sustainable agriculture and food security.     

TIME-INTENSITY MEASUREMENT OF MATRIX EFFECTS ON RETRONASAL AROMA PERCEPTION

A computerized time intensity procedure was used to evaluate the retronasal aroma intensity of vanillin and limonene in oil/water emulsion. The maximum perceived retronasal intensity of limonene was decreased as oil content in the emulsion increased, but temporal perception (time to maximum intensity and total duration of perception) was not affected. Total duration of vanillin retronasal aroma increased as oil content increased, and salivary flow rate had a significant effect on perception of vanillin aroma. These results suggest that time intensity measurements may be a valuable tool for evaluating the effects of matrix interactions on retronasal aroma intensity.     

Confusing tastes and smells: how odours can influence the perception of sweet and sour tastes

This study investigated the relationship between perception of an odour when smelled and the taste of a solution to which the odour is added as a flavorant. In Experiment 1 (E1) sweetness, sourness, liking and intensity ratings were obtained for 20 odours. Taste ratings were then obtained for sucrose solutions to which the odours had been added as flavorants. Certain odours were found to enhance tasted sweetness while others suppressed it. The degree to which an odour smelled sweet was the best predictor of the taste ratings. These findings were extended in Experiment 2 (E2), which included a second tastant, citric acid, and employed four odours from E1. The most sweet smelling odour, caramel, was found to suppress the sourness of citric acid and, as in E1, to enhance the sweetness of sucrose. Again, odours with low sweetness suppressed the sweetness of tasted sucrose. The study demonstrated that the effects of odours on taste perception are not limited to sweetness enhancement and apply to sour as well as sweet tastes. The overall pattern of results is consistent with an explanation of the taste properties of odours in terms of prior flavour-taste associations.     

FOCUSED DIFFERENCE TESTING FOR THE ASSESSMENT OF DIFFERENCES BETWEEN ORANGE JUICES MADE FROM ORANGE CONCENTRATE

Focused difference testing was applied to two orange juices prepared from frozen concentrate. Differences were noted between the juices for darkness of color, visual appearance of pulpiness, resistance to tongue movement, flavor by mouth, odor, overall taste and taste other than sweetness. The logic and approach of focused difference testing is discussed and contrasted with other sensory analytical techniques.