Flavor Aversion Research Articles

Identification of Compounds That Impact Consumer Flavor Liking of American-European Hazelnut Hybrids Using Nontargeted LC/MS Analysis.

American-European (Corylus americana × Corylus avellana) hazelnut hybrids are being developed for the Midwest-growing region of the United States. However, an inadequate understanding of the compounds that impact the consumer acceptance of hazelnuts limits breeding programs. Nontargeted liquid chromatography/mass spectrometry (LC/MS) chemical profiles of 12 roasted hybrid hazelnut samples and the corresponding consumer flavor liking scores were modeled by orthogonal partial least squares with good fit and predictive ability (R2Y > 0.9, Q2 > 0.9) to identify compounds that impact nut liking. The five most predictive compounds (1-5) were negatively correlated to flavor liking, selected as putative markers, purified by multidimensional preparative LC/MS, structurally elucidated (nuclear magnetic resonance, MS), quantified, and validated for sensory relevance. Compound 1 was identified as 1″-O-3'-b-glucofuranosyl-1'-O-1-b-glucofuranosyl-(2,6-dihydroxyphenyl)-ethan-4-one. Compounds 2 and 4 were identified as rotamers of 2-(3-hydroxy-2-oxoindolin-3-yl) acetic acid 3-O-6'-galactopyranosyl-2″-(2″oxoindolin-3″yl) acetate, whereas compounds 3 and 5 were identified as rotamers of 1″-O-1'-b-glucofuranosyl-9-O-6'-b-glucopyranosyl-2″-(2″-oxoindolin-3″yl) acetate. Sensory evaluation determined that all compounds were characterized by bitterness and/or astringency. The sensory threshold values of compounds 1-5 were determined to be below the concentrations reported in 91, 83, 41, 25, and 41% of all 12 hybrid hazelnut samples, respectively, indicating they contributed to aversive flavor attributes.

Food avoidance learning based on swimming in laboratory mice (Mus musculus)

Although it is now well documented that laboratory rats learn to avoid the flavored substance consumed immediately before running in activity wheels or swimming in water buckets, research on this activity-based flavor avoidance learning in other species is limited. Recently, running-based flavor avoidance learning has been demonstrated in laboratory mice by employing a method of resistance-to-habituation of neophobic reaction to novel food; mice that repeatedly experience running after encountering a novel food have a prolonged tendency to reject that food compared to control mice without paired running. The present article reports a series of attempts to obtain evidence of flavor avoidance learning based on swimming rather than running using this resistance-to-habituation method. Swimming-based flavor avoidance was clearly demonstrated in a differential conditioning paradigm; however, its demonstration in a simple conditioning paradigm requires a post-training choice test of the target food and another type of food. These results are likely due to the short swimming time (20 min) and the formation of weak flavor aversion.

A brainstem circuit for nausea suppression.

SUMMARYNausea is a discomforting sensation of gut malaise that remains a major clinical challenge. Several visceral poisons induce nausea through the area postrema, a sensory circumventricular organ that detects blood-borne factors. Here, we use genetic approaches based on an area postrema cell atlas to reveal inhibitory neurons that counteract nausea-associated poison responses. The gut hormone glucose insulinotropic peptide (GIP) activates area postrema inhibitory neurons that project locally and elicit inhibitory currents in nausea-promoting excitatory neurons through γ-aminobutyric acid (GABA) receptors. Moreover, GIP blocks behavioral responses to poisons in wild-type mice, with protection eliminated by targeted area postrema neuron ablation. These findings provide insights into the basic organization of nausea-associated brainstem circuits and reveal that area postrema inhibitory neurons are an effective pharmacological target for nausea intervention.

Establishment of Quality Standards for Raw Material of Cyanthillium cinereum (L.) H. Rob

Introduction: Cyanthillium cinereum has long been used in Thai traditional medicine. However, the standard information of C. cinereum raw material has not been established. Objective: This study aimed to present the standard information of C. cinereum raw material. Material and methods: The microscopic and macroscopic characteristics, the physicochemical analysis, and assays for bioactive contents of 24 samples of C. cinereum from different locations were evaluated to determine the standard information of the plant material. Results: The result showed the microscopic characteristic of pollens in all powder samples. No nicotine content in C. cinereum extract was observed with LC-MS; apigenin and luteolin were instead selected as markers. The observed average contents of apigenin and luteolin were 0.238 and 1.059 mg/g, respectively, using HPLC-UV. Nitrite and nitrate components in C. cinereum, which induced flavor aversion leading to smoking cessation, were determined. The average nitrite and nitrate content were 0.004 and 0.526 mg/kg in ethanol extract and 0.014 and 1.645 mg/kg in the water extract, respectively. The results did not substantiate the concern for high potassium content in C. cinereum materials for cardiovascular and renal patients where only traces were observed in all samples (3.113 and 20.898 mg/kg in ethanol and water extracts, respectively). Conclusion: The results can be used to establish the specification of C. cinereum, which will be useful for quality control of the raw material. This new information is crucial and can be utilized extensively prior to herbal product preparation.

Contextual control of the retardation of flavour aversion learning by preexposure to the unconditioned stimulus: Acquisition or retrieval deficit?

Two experiments, using rats as the subjects, and flavour aversion learning with an injection of lithium chloride (LiCl) as the unconditioned stimulus (US), examined the effects of a context shift between phases of the procedure on the retardation of learning produced by preexposure to the US. Experiment 1 showed that the US-preexposure effect (the reduction in the size of the conditioned aversion) was not attenuated when the animals were given both preexposure to the US and the conditioning procedure in a novel context but received the test phase in a different context (the home cages). Experiment 2 showed that, after degrading the injection cues–illness association by interpolating saline injections between LiCl preexposures, the US-preexposure effect was attenuated when there was a context shift between preexposure and conditioning, but that the context shift was without effect when it occurred between conditioning and test. These results are consistent with the proposal that US preexposure obtained in this procedure has its effect by interfering with the formation of the target association; they provide no support for the suggestion that the effect depends on interference at the test stage.

Identification of inhibitors of pinellic acid generation in whole wheat bread

Bitterness is a common aversive flavor attribute of foods associated with low consumer acceptance. Untargeted LC–MS flavoromic profiling was utilized to identify endogenous compounds that influence the generation of the bitter compound 9,12,13-trihydroxy-trans-10-octadecenoic acid (pinellic acid) during bread making. A diverse sample set of wheat germplasm was chemically profiled. The corresponding pinellic acid concentrations after dough formation were modeled by orthogonal partial least squares (OPLS) with good fit (R2Y = 0.8) and predictive ability (Q2 = 0.6). The most predictive feature (negatively correlated), postulated to interfere with the biosynthetic pathway, was identified as schaftoside, an apigenin di-C-glycoside. Recombination experiments involving the addition of schaftoside to flour prior to breadmaking resulted in a 26% decrease in pinellic acid formation and significantly lower perceived bitterness intensity in whole wheat bread. This work provides novel understanding of bitter generation pathways in wheat products and new strategies to improve flavor profiles and consumer acceptability.

Flavor Aversion Learning Based on Running: A Review

Wheel running establishes aversion in rats to a flavored solution consumed shortly before the running. Many studies have shown that this is a case of Pavlovian conditioning, in which the flavor and running respectively act as the conditioned stimulus (CS) and the unconditioned stimulus (US). The present article introduces some procedural variables of this running-based flavor aversion learning (FAL), including subjects, CS agents, US agents, and drive operations. This article also summarize various behavioral features of Pavlovian conditioning demonstrated in running-based FAL including the law of contiguity despite long-delay learning, extinction and spontaneous recovery, CS-preexposure effect, remote and proximal US-preexposure effects, degraded contingency effect, inhibitory learning by backward conditioning, stimulus overshadowing, associative blocking, and higher-order contextual control. Also reviewed are several hypotheses proposed for the underlying psychophysiological causes of running-based FAL (activation of mesolimbic dopamine system, gastrointestinal discomfort, motion sickness, energy expenditure, general stress, and anticipatory contrast). At the end of the article, we visit the question of most general interest about running-based FAL: why pleasurable activity of voluntary running yields aversive learning in rats.

Food avoidance learning based on voluntary wheel running in laboratory mice (Mus musculus)

Mice show a reluctance to eat unfamiliar food, when they first encounter it. This neophobic reaction is conventionally habituated by repeated trials: the mice gradually increase their consumption of the novel food. The new finding reported here is that the consumption remains low in mice that voluntarily run in activity wheels after the novel food access. This effect implies that running yields Pavlovian conditioned flavor aversion, which suppresses, otherwise increasing, consumption of the novel food. In the present research, the effect was demonstrated with a between-group design by pitting experimental mice receiving cheese-running paired treatment against cheese/running unpaired control mice (Experiment 1). The running-based food avoidance in mice was also shown in a differential conditioning paradigm, where one of two novel snacks (chocolate and marshmallow) was paired with running while the other was not, in non-deprived animals (Experiment 2 A) and food-deprived animals (Experiment 2B). These results concord with those previously reported in rats, indicating the generality of the phenomenon.

Social and anxiety-like behaviors contribute to nicotine self-administration in adolescent outbred rats

Both emotional and social traits interact with genetic factors to influence smoking behavior. We previously established a socially acquired nicotine intravenous self-administration model where social learning of a nicotine-associated odor cue reversed conditioned flavor aversion and promoted nicotine intake. In this study, we first phenotyped ~800 adolescent heterogeneous stock rats in open field, novel object interaction, social interaction, elevated plus maze, and marble burying behaviors. These rats were then phenotyped on socially acquired nicotine self-administration. We found 243 significant correlations between different behavioral tests. Principal component regression analysis found that ~10–20% of the variance in nicotine-related measures, such as intake during the first or the last three fixed-ratio sessions, the progressive ratio session, and reinstatement behavior, can be explained by variations in behavioral traits. Factors corresponding to social behavior and anxiety were among the strongest predictors of nicotine intake and reinstatement of nicotine-seeking behavior. We also found many sex differences in behavioral measures. These data indicated that the genetic diversity of this population, in combination with social behaviour and anxiety, are significant contributors to the divergent nicotine self-administration behavior and indicated a high probability of discovering sex-specific genetic mechanisms for nicotine intake in future genome-wide association studies.

Postnatal Exposure to Ethanol Increases Its Oral Acceptability to Adolescent Rats.

The aversive flavor of ethanol limits intake by many consumers. We asked whether intermittent consumption of ethanol increases its oral acceptability, using rats as a model system. We focused on adolescent rats because they (like their human counterparts) have a higher risk for alcohol overconsumption than do adult rats following experience with the drug. We measured the impact of ethanol exposure on 1) the oral acceptability of ethanol and surrogates for its bitter (quinine) and sweet (sucrose) flavor components in brief-access lick tests and 2) responses of the glossopharyngeal (GL) taste nerve to oral stimulation with the same chemical stimuli. During the exposure period, the experimental rats had access to chow, water and 10% ethanol every other day for 16 days; the control rats had access to chow and water over the same time period. The experimental rats consumed 7-14 g/day of 10% ethanol across the exposure period. This ethanol consumption significantly increased the oral acceptability of 3%, 6% and 10% ethanol, but had no impact on the oral acceptability of quinine, sucrose or NaCl. The ethanol exposure also diminished responses of the GL nerve to oral stimulation with ethanol, but not quinine, sucrose or NaCl. Taken together, these findings indicate that ethanol consumption increases the oral acceptability of ethanol in adolescent rats and that this increased oral acceptability is mediated, at least in part, by an exposure-induced reduction in responsiveness of the peripheral taste system to ethanol per se, rather than its bitter and sweet flavor components.

Assessment of zinc phosphide bait shyness and tools for reducing flavor aversions

Prairie voles (Microtus ochrogaster Wagner) cause extensive damage in agricultural, suburban, and urban environments. Control of these animals has historically relied on the use of anticoagulant rodenticides and zinc phosphide. However, shyness to zinc phosphide baits has reduced its efficacy. The aim of this study was to evaluate the factors involved in zinc phosphide bait shyness through preference testing. Baits were made using a rolled oat base and contained various combinations of the components of zinc phosphide baits such as lecithin, magnesium carbonate and known flavor modulators sodium cyclamate and zinc sulfate. Encapsulation of zinc phosphide was also tested as a potential means to mask undesirable flavor qualities of the compound. Consumption of test baits was measured in four day laboratory feeding trials. Results demonstrated that numerous components of current bait formulations serve as salient cues during conditioned aversions and therefore may contribute to bait shyness. Vole avoidance of zinc sulfate and sodium cyclamate revealed that these potential additives would not decrease bait shyness. Encapsulation of zinc phosphide may have masked some of the negative flavor cues and therefore should be considered in future bait development. This study suggests that, since voles are able to distinguish components of current bait formulations, varying composition of zinc phosphide baits between applications may serve to reduce bait shyness.

Social learning promotes nicotine self-administration by facilitating the extinction of conditioned aversion in isogenic strains of rats

Both social environment and genetic factors are critical for smoking initiation and nicotine addiction. We reported that rats developed conditioned flavor (i.e., taste and odor) aversion to intravenously self-administered (IVSA) nicotine, and that social learning promoted nicotine IVSA with flavor cues. We thus tested the hypothesis that socially acquired nicotine IVSA is a heritable trait by using female rats of six inbred strains and six F1 hybrids. Each strain was tested for 10 daily IVSA sessions. We found that the intake of nicotine (15 and 30 μg/kg/inf) varied among these strains by 33.7–56.6-fold. The heritability of nicotine intake was estimated to be 0.54–0.65. Further, there was a strong correlation in nicotine intake (R2 = 0.85, p < 0.0001) between the two nicotine doses. Another cohort of rats was given three daily IVSA sessions followed by five sessions that tested conditioned flavor aversion. Nicotine intake was highly correlated with the extinction of the conditioned aversion (R2 = 0.58, p < 0.005). These data showed that nicotine intake in the socially acquired nicotine self-administration model is controlled by genetic factors and that the role of social learning is likely in facilitating the extinction of conditioned aversive response to nicotine.

Perceptual learning in flavor preference conditioning: Restricting generalization of acquired preferences between flavors

Two experiments with rats investigated perceptual learning using a conditioned preference procedure. Experiment 1 used a between-subject procedure in which rats received either intermixed preexposure (AX, BX, AX, BX…) or blocked preexposure (AX, AX…, BX, BX…) to flavor compounds before a conditioned preference was established to AX by pairing it with sucrose. During a test, rats given intermixed preexposure showed a greater preference for AX over BX than those given blocked preexposure. Experiment 2 showed that after intermixed preexposure to AX and BX, and a block of preexposure to CX, a preference established to AX was less likely to generalize to BX than to CX. These results represent the first demonstration of the impact of the schedule of preexposure on perceptual learning using a flavor preference procedure, and they parallel those previously observed using flavor aversion procedures.

Long-Term Alcohol Drinking Reduces the Efficacy of Forced Abstinence and Conditioned Taste Aversion in Crossed High-Alcohol-Preferring Mice.

Negative outcomes of alcoholism are progressively more severe as the duration of problem of alcohol use increases. Additionally, alcoholics demonstrate tendencies to neglect negative consequences associated with drinking and/or to choose to drink in the immediate presence of warning factors against drinking. The recently derived crossed high-alcohol-preferring (cHAP) mice, which volitionally drink to heavier intoxication (as assessed by blood ethanol [EtOH] concentration) than other alcohol-preferring populations, as well as spontaneously escalating their intake, may be a candidate to explore mechanisms underlying long-term excessive drinking. Here, we hypothesized that an extended drinking history would reduce the ability of 2 manipulations (forced abstinence [FA] and conditioned taste aversion [CTA]) to attenuate drinking. Experiment 1 examined differences between groups drinking for either 14 or 35days, half of each subjected to 7days of FA and half not, to characterize the potential changes in postabstinence drinking resulting from an extended drinking history. Experiment 2 used a CTA procedure to assess stimulus specificity of the ability of an aversive flavorant to decrease alcohol consumption. Experiment 3 used this taste aversion procedure to assess differences among groups drinking for 1, 14, or 35days in their propensity to overcome this aversion when the flavorant was mixed with either EtOH or water. Experiment 1 demonstrated that although FA decreased alcohol consumption in mice with a 14-day drinking history, it failed to do so in mice drinking alcohol for 35days. Experiment 2 showed that the addition of a flavorant only suppressed alcohol drinking if an aversion to the flavorant was previously established. Experiment 3 demonstrated that an extended drinking history expedited extinction of suppressed alcohol intake caused by a conditioned aversive flavor. These data show that a history of long-term drinking in cHAP mice attenuates the efficacy of interventions that normally reduce drinking. Analogous to alcoholics who may encounter difficulties in limiting their intake, cHAP mice with long drinking histories are relatively insensitive to both abstinence and signals of harmful consequences. We propose that the cHAP line may be a valid model for adaptations that occur following the extended heavy alcohol drinking.

Behavior of suckling pigs supplemented with an encapsulated iron oral formula

Parental iron supplementation in neonatal piglets is a routine management to solve anemia but may cause welfare problems. The aim of this study was to assess behavior responses generated after oral or parenteral iron supplementation. Parenteral supplementation consisted of 200 mg of iron dextran intramuscular. The novel oral iron supplement was a combination of encapsulated nonheme/heme iron (252 mg of total iron) delivered orally. Two litters formed by 11, 2-day-old piglets were assigned to each treatment. The litters were video recorded for 6 hours, 3 before and 3 after iron supplementation. Scan sampling every 3 minutes was used to register the occurrence of 7 behaviors. The highest overall percentage of time was allocated to resting, 46.5% (167.4 minutes) for the parenteral group and 42.4% (152.6 minutes) for the oral group; and suckling 24.6% (88.6 minutes) for the parenteral group and 27.8% (100.1 minutes) for the oral group. Resting time was higher after 1 hour of iron supplementation in the parenteral group 51.9% (31.1 minutes) vs. 33.8% (19.7 minutes). In conclusion, the oral iron supplementation resulted in a higher behavioral disruption in neonatal pigs, probably associated to increased handling time and aversive flavor of the supplement.

Socially acquired nicotine self-administration with an aversive flavor cue in adolescent female rats.

Establishing a behavioral model for the effect of social environment on nicotine intake in rodents facilitates the investigation of molecular mechanisms critical for the interaction between social environment and cigarette smoking. Our main objective was to test the hypothesis that nicotine is the primary reinforcer in the socially acquired nicotine intravenous self-administration (IVSA) model by using an aversive flavor cue. Adolescent female rats were placed in operant conditioning chambers equipped with two lickometers. Operant licking triggered concurrent deliveries of a flavor (i.e., taste and odor) cue containing either quinine or saccharin and an i.v. infusion (30μg/kg nicotine or saline). An audiovisual cue was provided for some groups of rats. A second rat that did not receive nicotine was placed in the operant conditioning chambers to provide either a neutral or an inducing (i.e., by consuming the flavored solution) social environment. These two rats were separated by a divider that allowed orofacial interactions. Rats acquired stable nicotine IVSA with either the aversive or the appetitive flavor cue in the inducing social environment, and obtained similar amounts of infusions. The neutral social environment did not support nicotine IVSA with either cue. The audiovisual cue per se did not support nicotine IVSA but enhanced nicotine intake. Nicotine increased the number of concurrent nose pokes by the two rats into the center divider, a measure of social interaction. Despite its aversive effects, nicotine is the primary reinforcer for the operant responses in the socially acquired nicotine IVSA model.

Naloxone impairs concurrent but not sequential flavor aversion: Resorting to a flexible/explicit learning.

The role of opiate systems has been extensively studied in relation to learning and memory. Naloxone (Nx), an opiate antagonist, was administrated in concurrent (Experiment 1) and sequential (Experiment 2) flavor aversion learning (FAL) tasks. The outcomes demonstrate that Nx impairs the acquisition of concurrent but not sequential FAL. In the concurrent learning (7 trials), both control (vehicle) and Nx2 (treated with Nx only on the first 2 days) groups learned the task. Furthermore, these 2 groups retained the learning in a discrimination test without drug administration (Day 8) but failed a reversal test (Day 9). In contrast, the Nx group (7 trials with Nx) showed no concurrent learning but correctly performed the discrimination test (Day 8) and, critically, the reversal test. These results suggest that Nx blocks concurrent (implicit) learning in these experiments but induces animals to resort to new strategies that are flexible, a characteristic of explicit learning.

Lesions of the central nucleus of the amygdala only impair flavor aversion learning in the absence of olfactory information

The amygdala is considered a crucial brain nucleus in different modalities of aversive conditioning, including flavor aversion learning (FAL). The importance attributed to the amygdala and its subnuclei has frequently depended on the different stimuli and procedures used in FAL tasks. In this study, FAL was impaired only in animals that had lesions in the central nucleus of the amygdala (CeA) area and also had their olfactory bulbs removed. However, this task was learned by neurologically intact animals, bulbectomized animals, and rats with lesions exclusively centered in the CeA area alone. These results suggest that the CeA area may be relevant in gustatory-gut associative learning but not in FAL, in which the olfactory system may counteract the deficit produced in taste-visceral convergence.

Within-compound associations mediate augmented flavor aversions

Five experiments with rat subjects explored flavor-aversion conditioning in the A+/AX+ design. In Experiment 1, odor preconditioning prior to taste+odor compound conditioning significantly strengthened a taste aversion, and taste preconditioning prior to taste+odor conditioning significantly augmented an odor aversion. Experiments 2A and 2B eliminated the alternative explanation that the augmentation effect was produced by differential exposure to the compound during conditioning. Next, we examined a possible mechanism of these augmentation effects by employing post-conditioning extinction of the augmenting stimulus (CS A); previous research had also used this technique to explore augmentation, but those studies yielded conflicting results. In both Experiments 3A and 3B, post-conditioning extinction of CS A led to a significant decrease in the strength of the augmented aversion to CS X, regardless of stimulus modality or stimulus salience. Collectively, these results suggest augmented flavor aversions are produced via within-compound associations between CS A and CS X.

Effects of dietary choline availability on latent inhibition of flavor aversion learning

ObjectiveIt has been previously reported that dietary choline supplementation might affect latent inhibition (LI) using a conditioned suppression procedure in rats. We have assessed the effect of dietary choline on LI of flavor aversion learning.MethodAdult male Wistar rats received a choline supplemented (5 g/kg), deficient (0 g/kg), or standard (1.1 g/kg) diet for 3 months. After this supplementation period, all rats went through a conditioned taste aversion (CTA) procedure, half of them being pre-exposed to the conditioned stimulus before the conditioning.ResultsThe results indicated that choline deficiency prevents LI of conditioned flavor aversion to cider vinegar (3%) induced by a LiCl (0.15 M; 2% body weight) intraperitoneal injection, while choline supplementation enhances CTA leading to slower extinction.DiscussionThe role of the brain systems modulating attentional processes is discussed.