A lubrication analysis of pharyngeal peristalsis: Application to flavour release

After eating a liquid or a semi-liquid food product, a thin film responsible for the dynamic profile of aroma release coats the pharyngeal mucosa. The aim of this article was to analyse the fluid mechanics of pharyngeal peristalsis and to develop a simple biomechanical model in order to understand the role of saliva and food bolus viscosity on the coating of pharyngeal mucosa. We began by analysing the physiology and the biomechanics of swallowing in order to determine relevant model assumptions. This analysis of the literature clarified the types of mechanical solicitations applied on the food bolus. Moreover, we showed that the pharyngeal peristalsis in the most occluded region is equivalent to a forward roll coating process, the originality of which is lubrication by a film of saliva. A model based on the lubrication theory for Newtonian liquids was developed in dimensionless form. The parametric study showed the strong influence of relative saliva thickness on the food bolus coating. A specific experimental device was designed that confirms the model predictions. Two sets of conditions that depend on the relative thickness of saliva were distinguished. The first is characterised by a relatively thin film of saliva: food bolus viscosity has a strong impact on mucosa coating. These phenomena are well represented by the model developed here. The second is obtained when the saliva film is relatively thick: hydrodynamic mixing with saliva, interdiffusion or instabilities may govern mucosa coating. Finally, these results were extrapolated to determine the influence of food bolus viscosity on the dynamic profile of flavour release according to physiological parameters.     

An experimental model to investigate the biomechanical determinants of pharyngeal mucosa coating during swallowing

The development of innovative experimental approaches is necessary to gain insights in the complex biomechanics of swallowing. In particular, unraveling the mechanisms of formation of the thin film of bolus coating the pharyngeal mucosa after the ingestion of liquid or semi-liquid food products is an important challenge, with implication in dysphagia treatment and sensory perceptions.The aim here is to propose an original experimental model of swallowing (i) to simulate the peristaltic motions driving the bolus from the oral cavity to the esophagus, (ii) to mimic and vary complex physiological variables of the pharyngeal mucosa (lubrication, deformability and velocity) and (iii) to measure the thickness and the composition of the coatings resulting from bolus flow. Three Newtonian glucose solutions were considered as model food boli, through sets of experiments covering different ranges of each physiological parameter mimicked.The properties of the coatings (thickness and dilution in saliva film) were shown to depend significantly on the physical properties of food products considered (viscosity and density), but also on physiological variables such as lubrication by saliva, velocity of the peristaltic wave, and to a lesser extent, the deformability of the pharyngeal mucosa.The biomechanical peristalsis simulator developed here can contribute to unravel the determinants of bolus adhesion on pharyngeal mucosa, necessary both for the design of alternative food products for people affected by swallowing disorders, and for a better understanding of the dynamic mechanisms of aroma perception.     

An optimization model for mastication and swallowing in mammals.

Mammalian mastication is a process combining simultaneous food comminution and lubrication. The initiation of swallowing, which is voluntary, has been thought to depend on separate thresholds for food particle size and for particle lubrication. Instead of this duality, we suggest that swallowing is initiated when it is sensed that a batch of food particles is binding together under viscous forces so as to form a bolus. Bolus formation ensures that when the food mass is swallowed, it will pass the pharyngeal region safely without risk of inhaling small particles into the lower respiratory tract. Crucial for bolus formation is food particle size reduction by mastication. This allows the tongue to pack particles together tightly by pressure against the hard palate. A major function of salivation is to fill the gradually reducing spaces between particles, so increasing viscous cohesion and promoting bolus formation. If swallowing is delayed, excessive saliva floods the bolus, separating particles and reducing cohesion. Swallowing then becomes more precarious. Our model suggests that there is an optimum moment for a mammal to swallow, defined in terms of a peak cohesive force between food particles. The model is tested on human mastication with two foods, brazil nut and raw carrot, which have very different particle size breakdown rates. The peak cohesive force is much greater with brazil nuts but both foods are predicted to be swallowed after similar numbers of chews despite the very different food particle size reductions achieved at that stage. The predicted number of chews to swallow is in broad agreement with published data.     

Correspondence between food consistency and suprahyoid muscle activity, tongue pressure, and bolus transit times during the oropharyngeal phase of swallowing.

This study aimed to evaluate the effects of food texture and viscosity on the swallowing function by measuring tongue pressure and performing a videofluorographic (VF) examination. Eleven normal adults were recruited for this study. Test foods with different consistencies and liquid contents, i.e., a half-solid nutrient made of 0.8 and 1.5% agar powder, syrup, and a liquid containing 40 wt/vol% barium sulfate, were swallowed, and the anterior (AT) and posterior tongue pressures (PT) and electromyographic (EMG) activity of the suprahyoid muscles were recorded, together with VF images. The timing of each event obtained from EMG, tongue pressure, and VF recordings was measured and then compared. We found that the AT and PT activity patterns were similar and showed a single peak. The peak, area, and time duration of all of the variables for AT and PT and EMG burst increased with increasing hardness of the bolus. The onset of the EMG burst always preceded those of the AT and PT activities, while there were no significant differences in peak and offset times among EMG burst, AT, and PT. Total swallowing time and oral ejection time were significantly longer during the swallowing of 1.5% agar than any other boluses, while pharyngeal transit time and clearance time were significantly longer during the swallowing of syrup, which was as hard as the liquid, but showed a higher viscosity than the liquid. The results suggested that the major effects of food hardness were to delay oral ejection time, which strongly delays total swallowing time. In addition, pharyngeal bolus transit is not dependent on the hardness of food but on its viscosity.     

Electromyographic activity from human laryngeal, pharyngeal, and submental muscles during swallowing.

The durations and temporal relationships of electromyographic activity from the submental complex, superior pharyngeal constrictor, cricopharyngeus, thyroarytenoid, and interarytenoid muscles were examined during swallowing of saliva and of 5- and 10-ml water boluses. Bipolar, hooked-wire electrodes were inserted into all muscles except for the submental complex, which was studied with bipolar surface electrodes. Eight healthy, normal, subjects produced five swallows of each of three bolus volumes for a total of 120 swallows. The total duration of electromyographic activity during the pharyngeal stage of the swallow did not alter with bolus condition; however, specific muscles did show a volume-dependent change in electromyograph duration and time of firing. Submental muscle activity was longest for saliva swallows. The interarytenoid muscle showed a significant difference in duration between the saliva and 10-ml water bolus. Finally, the interval between the onset of laryngeal muscle activity (thyroarytenoid, interarytenoid) and of pharyngeal muscle firing patterns (superior pharyngeal constrictor onset, cricopharyngeus offset) decreased as bolus volume increased. The pattern of muscle activity associated with the swallow showed a high level of intrasubject agreement; the presence of somewhat different patterns among subjects indicated a degree of population variance.     

Mechanistic mathematical model for in vivo aroma release during eating of semiliquid foods

The paper describes a mechanistic mathematical model for aroma release in the oropharynx to the nasal cavity during food consumption. The model is based on the physiology of the swallowing process and is validated with atmospheric pressure chemical ionization coupled with mass spectrometry measurements of aroma concentration in the nasal cavity of subjects eating flavored yogurt. The study is conducted on 3 aroma compounds representative for strawberry flavor (ethyl acetate, ethyl butanoate, and ethyl hexanoate) and 3 panelists. The model provides reasonably accurate time predictions of the relative aroma concentration in the nasal cavity and is able to simulate successive swallowing events as well as imperfect velopharyngeal closure. The most influent parameters are found to be the amount of the residual product in the pharynx and its contact area with the air flux, the volume of the nasal cavity, the equilibrium air/product partition coefficient of the volatile compound, the breath airflow rate, as well as the mass transfer coefficient of the aroma compound in the product, and the amount of product in the mouth. This work constitutes a first step toward computer-aided product formulation by allowing calculation of retronasal aroma intensity as a function of transfer and volatility properties of aroma compounds in food matrices and anatomophysiological characteristics of consumers.     

Retro-nasal aroma release is correlated with variations in the in-mouth air cavity volume after empty deglutition

We hypothesized that interindividual differences in motor activities during chewing and/or swallowing were determining factors for the transfer of volatile aroma from the in-mouth air cavity (IMAC) toward the olfactory mucosa. In our first experiment, we looked for changes in IMAC volume after saliva deglutition in 12 healthy subjects. The mean IMAC volume was measured after empty deglutition using an acoustic pharyngometer device. Based on the time course of the IMAC volume after swallowing, we discerned two groups of subjects. The first group displayed a small, constant IMAC volume (2.26 mL ±0.62) that corresponded to a high tongue position. The second group displayed a progressive increase in IMAC (from 6.82 mL ±2.37 to 22.82 mL ±3.04) that corresponded to a progressive lowering of the tongue to its resting position. In our second experiment, we investigated the relationship between IMAC volume changes after deglutition and the level of aroma release at the nostril. For this purpose, the release of menthone was measured at the nostril level in 25 subjects who consumed similar amounts of a mint tablet. The subjects were separated into two groups corresponding to two levels of menthone release: high (H) and low (L). The mean volume of IMAC was measured during and after empty deglutition. Group H displayed a small, constant amplitude of IMAC volume change after deglutition, while Group L displayed a progressive increase in IMAC. It is likely that Group H continuously released the aroma through the veloglossal isthmus as the mint was consumed, while Group L trapped the aroma in the oral cavity and then released it into the nasal cavity upon swallowing. These results show that the in vivo aroma release profile in humans depends closely on the different motor patterns at work during empty deglutition.     

Assessment of intraluminal impedance for the detection of pharyngeal bolus flow during swallowing in healthy adults

Intraluminal impedance, a nonradiological method for assessing bolus flow within the gut, may be suitable for investigating pharyngeal disorders. This study evaluated an impedance technique for the detection of pharyngeal bolus flow during swallowing. Patterns of pharyngoesophageal pressure and impedance were simultaneously recorded with videofluoroscopy in 10 healthy volunteers during swallowing of liquid, semisolid, and solid boluses. The timing of bolus head and tail passage recorded by fluoroscopy was correlated with the timing of impedance drop and recovery at each recording site. Bolus swallowing produced a drop in impedance from baseline followed by a recovery to at least 50% of baseline. The timing of the pharyngeal and esophageal impedance drop correlated with the timing of the arrival of the bolus head. In the pharynx, the timing of impedance recovery was delayed relative to the timing of clearance of the bolus tail. In contrast, in the upper esophageal sphincter (UES) and proximal esophagus, the timing of impedance recovery correlated well with the timing of clearance of the bolus tail. Impedance-based estimates of pharyngoesophageal bolus clearance time correlated with true pharyngoesophageal bolus clearance time. Patterns of intraluminal impedance recorded in the pharynx during bolus swallowing are therefore more complex than those in the esophagus. During swallowing, mucosal contact between the tongue base and posterior pharyngeal wall prolongs the duration of pharyngeal impedance drop, leading to overestimation of bolus tail timing. Therefore, we conclude that intraluminal impedance measurement does not accurately reflect the bolus transit in the pharynx but does accurately reflect bolus transit across the UES and below.     

Respiration during feeding on solid food: alterations in breathing during mastication, pharyngeal bolus aggregation, and swallowing.

During feeding, solid food is chewed and propelled to the oropharynx, where the bolus gradually aggregates while the larynx remains open and breathing continues. The aggregated bolus in the valleculae is exposed to respiratory airflow, yet aspiration is rare in healthy individuals. The mechanism for preventing aspiration during bolus aggregation is unclear. One possibility is that alterations in the pattern of respiration during feeding could help prevent inhalation of food from the pharynx. We hypothesized that respiration was inhibited during bolus aggregation in the valleculae. Videofluorography was performed on 10 healthy volunteers eating solid foods with barium. Respiration was monitored concurrently with plethysmography and nasal air pressure. The timing of events during mastication, food transport, pharyngeal bolus aggregation, and swallowing were measured in relation to respiration. Respiratory cycle duration decreased during chewing (P < 0.001) but increased with swallowing (P < 0.001). During 66 recordings of vallecular bolus aggregation, there was inspiration in 8%, expiration in 41%, a pause in breathing in 17%, and multiple phases (including inspiration) in 35%. Out of 98 swallows, 47% started in the expiratory phase and 50% started during a pause in breathing, irrespective of bolus aggregation in the valleculae. Plethysmography was better than nasal manometry for determining the end of active expiration during feeding and swallowing with solid food. The hypothesis is rejected in that respiration was not inhibited during bolus aggregation. These findings suggest that airflow through the pharynx does not have a role in preventing aspiration during bolus aggregation in the oropharynx.     

Mechanisms of airway protection during retching,vomiting, and swallowing

We investigated the mechanisms of airway protection and bolus transport during retching and vomiting by recording responses of the pharyngeal, laryngeal, and hyoid muscles and comparing them with responses during swallowing and responses of the gastrointestinal tract. Five dogs were chronically instrumented with electrodes on the striated muscles and strain gauges on smooth muscles. Retching and vomiting were stimulated by apomorphine (5-10 ug/kg iv). During retching, the hyoid and thyroid descending and laryngeal abductor muscles were activated; between retches, the hyoid, thyroid, and pharyngeal elevating, and laryngeal adductor muscles were activated. Vomiting always occurred during the ascending phase of retching and consisted of three sequential phases of hyoid and pharyngeal muscle activation culminating in simultaneous activation of all recorded elevating and descending laryngeal, hyoid, and pharyngeal muscles. Retrograde activation of esophagus and pharyngeal muscles occurred during the later phases, and laryngeal adductor was maximally activated in all phases of the vomit. During swallowing, the laryngeal adductor activation was followed immediately by brief activation of the laryngeal abductor. We concluded that retching functions to mix gastric contents with refluxed intestinal secretions and to impart an orad momentum to the bolus before vomiting. During retches, the airway is protected by glottal closure, and between retches, it is protected by ascent of the larynx and closure of the upper esophageal sphincter. The airway is protected by maximum glottal closure during vomiting. During swallowing, the airway is protected by laryngeal elevation and glottal closure followed by brief opening of the glottis, which may release subglottal pressure expelling material from the laryngeal vestibule.     

Adaptation of swallowing hyo-laryngeal kinematics is distinct in oral vs. pharyngeal sensory processing

Before a bolus is pushed into the pharynx, oral sensory processing is critical for planning movements of the subsequent pharyngeal swallow, including hyoid bone and laryngeal (hyo-laryngeal) kinematics. However, oral and pharyngeal sensory processing for hyo-laryngeal kinematics is not fully understood. In 11 healthy adults, we examined changes in kinematics with sensory adaptation, sensitivity shifting, with oropharyngeal swallows vs. pharyngeal swallows (no oral processing), and with various bolus volumes and tastes. Only pharyngeal swallows showed sensory adaptation (gradual changes in kinematics with repeated exposure to the same bolus). Conversely, only oropharyngeal swallows distinguished volume differences, whereas pharyngeal swallows did not. No taste effects were observed for either swallow type. The hyo-laryngeal kinematics were very similar between oropharyngeal swallows and pharyngeal swallows with a comparable bolus. Sensitivity shifting (changing sensory threshold for a small bolus when it immediately follows several very large boluses) was not observed in pharyngeal or oropharyngeal swallowing. These findings indicate that once oral sensory processing has set a motor program for a specific kind of bolus (i.e., 5 ml water), hyo-laryngeal movements are already highly standardized and optimized, showing no shifting or adaptation regardless of repeated exposure (sensory adaptation) or previous sensory experiences (sensitivity shifting). Also, the oral cavity is highly specialized for differentiating certain properties of a bolus (volume) that might require a specific motor plan to ensure swallowing safety, whereas the pharyngeal cavity does not make the same distinctions. Pharyngeal sensory processing might not be able to adjust motor plans created by the oral cavity once the swallow has already been triggered.     

Kinematics of the pharyngeal jaws during feeding in Oreochromis niloticus (Pisces,Perciformes)

Cichlids possess a complex pharyngeal jaw apparatus, the osteological components of which are two upper pharyngeal jaws, articulating with the neurocranial base, and a single lower pharyngeal jaw. Quantitative cinera-diography revealed that pharyngeal food processing in Oreochromis niloticus involves transport, mastication, and swallowing, effected by cyclical pharyngeal jaw movements. Transport and swallowing occur by simultaneous retractions of both upper pharyngeal jaws. Food reduction (mastication) is effected by lower jaw elevation (compression) and protraction (shear) during upper jaw retraction. Each movement cycle contains a transport, reduction, and swallowing component, although their relative importance may vary within a feeding sequence. The upper and lower pharyngeal jaws show opposite anteroposterior movements during most of the cycle. Variations in the amplitudes and the durations of the different movement components reflect the consistency and the size of the food.     

Influence of bite force and tongue pressure on oro-pharyngeal residue in the elderly

Objective: To investigate the influence of maximal bite force, maximal tongue pressure, number of mastications and swallowing on the oro‐pharyngeal residue in the elderly. Background: Oro‐pharyngeal residue in the elderly is an indication of dysphagia. Pharyngeal residue is especially critical as it may cause aspiration pneumonia, which is one of the major causes of death in elderly. Materials and methods: Videofluorographic recordings were performed on 14 elderly volunteers (six males, eight females, age range 65–93 years) without any history or symptoms of dysphagia. The subjects were instructed to consume 9 g of barium containing bread in two manners; free mastication and swallow (FMS: masticate and swallow freely), and limited mastication and swallow (LMS: swallow once after 30 chewing actions). The amount of oral and pharyngeal residue was evaluated using a 4‐point rating scale. Maximal occlusal force was measured by a pressure sensitive sheet, and maximal tongue pressure using a handy probe. Multiple regression analysis was performed to examine the influence of these items on the amount of oral and pharyngeal residue in FMS and LMS. Results: In FMS, age was found to be a factor which increased oral residue (p = 0.053), and the number of swallowing (p = 0.017) and the state of the prosthesis (p = 0.030) reduced the pharyngeal residue. In LMS, tongue pressure was a factor which reduced oral residue (p = 0.015) and increased pharyngeal residue (p = 0.008). Conclusion: It is suggested that in the elderly tongue pressure contributed to propulsion of the food bolus from oral cavity into the pharynx, and multiple swallowing contributed to the reduction in the amount of pharyngeal residue.     

The coordination and interaction between respiration and deglutition in young pigs

The anatomical pathways for inspired air and ingested food cross in the pharynx of mammals, implying that breathing and swallowing must be separated either in space or in time. In this study we investigated the time relationship between swallowing and respiration in young pigs, as a model for suckling mammals. Despite the high morphological position of the larynx in young mammals, allowing liquid to pass in food channels lateral to the larynx, respiration and swallowing are not wholly independent events. Although, when suckling on a veterinary teat, the swallows occurred at various points in the respiratory cycle, there was always a period of apnea associated with the swallow. Finally, an increase in the viscosity of the milk altered this coordination, changing respiratory cycle length and also restricting the relative rate at which swallows occurred in some parts of the respiratory cycle. These results suggest that the subsequent changes in respiratory activity at weaning, associated with passage of a solid bolus over the larynx, is preceded by the ability of the animal to alter coordination between respiration and swallowing for a liquid bolus. Accepted: 29 September 1997     

Upper esophageal sphincter impedance as a marker of sphincter opening diameter

The measurement of the physical extent of opening of the upper esophageal sphincter (UES) during bolus swallowing has to date relied on videofluoroscopy. Theoretically luminal impedance measured during bolus flow should be influenced by luminal diameter. In this study, we measured the UES nadir impedance (lowest value of impedance) during bolus swallowing and assessed it as a potential correlate of UES diameter that can be determined nonradiologically. In 40 patients with dysphagia, bolus swallowing of liquids, semisolids, and solids was recorded with manometry, impedance, and videofluoroscopy. During swallows, the UES opening diameter (in the lateral fluoroscopic view) was measured and compared with automated impedance manometry (AIM)-derived swallow function variables and UES nadir impedance as well as high-resolution manometry-derived UES relaxation pressure variables. Of all measured variables, UES nadir impedance was the most strongly correlated with UES opening diameter. Narrower diameter correlated with higher impedance (r = -0.478, P < 0.001). Patients with <10 mm, 10-14 mm (normal), and ≥ 15 mm UES diameter had average UES nadir impedances of 498 ± 39 Ohms, 369 ± 31 Ohms, and 293 ± 17 Ohms, respectively (ANOVA P = 0.005). A higher swallow risk index, indicative of poor pharyngeal swallow function, was associated with narrower UES diameter and higher UES nadir impedance during swallowing. In contrast, UES relaxation pressure variables were not significantly altered in relation to UES diameter. We concluded that the UES nadir impedance correlates with opening diameter of the UES during bolus flow. This variable, when combined with other pharyngeal AIM analysis variables, may allow characterization of the pathophysiology of swallowing dysfunction.     

Role of physical bolus properties as sensory inputs in the trigger of swallowing

Background

Swallowing is triggered when a food bolus being prepared by mastication has reached a defined state. However, although this view is consensual and well supported, the physical properties of the swallowable bolus have been under-researched. We tested the hypothesis that measuring bolus physical changes during the masticatory sequence to deglutition would reveal the bolus properties potentially involved in swallowing initiation.

Methods

Twenty normo-dentate young adults were instructed to chew portions of cereal and spit out the boluses at different times in the masticatory sequence. The mechanical properties of the collected boluses were measured by a texture profile analysis test currently used in food science. The median particle size of the boluses was evaluated by sieving. In a simultaneous sensory study, twenty-five other subjects expressed their perception of bolus texture dominating at any mastication time.

Findings

Several physical changes appeared in the food bolus as it was formed during mastication: (1) in rheological terms, bolus hardness rapidly decreased as the masticatory sequence progressed, (2) by contrast, adhesiveness, springiness and cohesiveness regularly increased until the time of swallowing, (3) median particle size, indicating the bolus particle size distribution, decreased mostly during the first third of the masticatory sequence, (4) except for hardness, the rheological changes still appeared in the boluses collected just before swallowing, and (5) physical changes occurred, with sensory stickiness being described by the subjects as a dominant perception of the bolus at the end of mastication.

Conclusions

Although these physical and sensory changes progressed in the course of mastication, those observed just before swallowing seem to be involved in swallowing initiation. They can be considered as strong candidates for sensory inputs from the bolus that are probably crucially involved in the triggering of swallowing, since they appeared in boluses prepared in various mastication strategies by different subjects.     

Impact of swallowing on the dynamics of aroma release and perception during the consumption of alcoholic beverages

The consumption protocol used during alcoholic beverage tasting may affect aroma perception. We used an integrated approach combining sensory analysis and physicochemistry to investigate the impact of swallowing on aroma release and perception. A panel of 10 persons evaluated the dynamics of aroma perception during the consumption of a commercial flavored vodka, using the method of temporal dominance of sensations. Two protocols (spitting out or swallowing of the product) were tested. Nosespace analysis was simultaneously carried out by proton transfer reaction mass spectrometry to evaluate aroma release in the nasal cavity. Comparison of the results obtained with the 2 protocols highlighted significant differences in both the perception and the release of aroma: the swallowing of the product resulted in more complex perceptions but decreased the dominance rates of aromatic attributes. Ethanol perception also had an impact when the product was swallowed. Aroma release data partly accounted for the differences in perception, particularly as concerned ethanol release. The time at which dominance appears as well as the dominance duration of some attributes can be related to some temporal parameters of release data. But the lack of knowledge concerning the variety and complexity of mechanisms continues to limit our understanding of relationship between aroma release and perception.     

Laryngeal activity during upright vs. supine swallowing.

Previous investigations of human pharyngeal muscle activation patterns during swallowing found a relatively invariant muscle activation onset sequence in the upright position. However, different gravitational forces influence a liquid bolus when supine and could modify the central timing control of laryngeal airway protection during swallowing. The purpose of this study was to determine whether laryngeal muscle onset timing during swallowing differed between the supine and upright positions. Nine subjects performed six swallowing trials with a 2-ml water bolus in each position. Simultaneous electromyographic recordings were obtained from the submental complex (SMC) and the right and left thyroarytenoid (TA) muscles. Regardless of body position, the timing, amplitude, and duration of the TA muscles did not vary relative to the SMC. Therefore, the sequence of TA muscle activation relative to the SMC during swallowing appeared unaffected by gravitational influences.     

Salivary β-glucosidase as a direct factor influencing the occurrence of halitosis

β-Glucosidases are enzymes present in all living organisms, playing a pivotal role in diverse biological processes. These enzymes cleave β-glycosidic bonds between carbohydrates, or between a carbohydrate and a non-carbohydrate moiety, which may result in the liberation of volatile aglycones. Released compounds execute diverse physiological roles, while the industry takes advantage of exogenously added β-glucosidases for aroma enrichment during food and beverage production. β-Glucosidase enzymatic activity has been reported in human saliva and given the fact that these enzymes are involved in aroma release, we investigated here the correlation between β-glucosidase activity in human saliva and the occurrence of halitosis. Measurement of salivary enzyme activity of 48 volunteers was performed using p-nitrophenyl-β-d-glucopyranoside as substrate. Each volunteer was clinically evaluated by a dental surgeon and clinical and laboratorial data were statistically analyzed. Gas-chromatography of saliva headspace allowed the analysis of the direct role of exogenous β-glucosidase on aromatic /volatile profile of saliva samples. The data demonstrated a positive correlation between halitosis and enzymatic activity, suggesting that the enzyme exerts a direct role in the occurrence of bad breath. Gas-chromatography analysis demonstrated that exogenously added enzyme led to the alteration of volatile organic content, confirming a direct contribution of β-glucosidase activity on saliva volatile compounds release. Although halitosis is a multifactorial condition, the complete understanding of all governing factors may allow the development of more effective treatment strategies. Such studies may pave the way to the use of β-glucosidase inhibitors for halitosis clinical management.