1

The Effect of Specializing Diets On Affective Facial Responses in Primates

Cassandra Dawson 0599587 PSYC 4050 - L. Parker

Monday November 26th 2012 University of Guelph

2 Abstract Based on the study by Steiner, Glaser, Hawilo, and Berridge (2000) affective responses to bitter and sweet tasting stimuli can be quantified as an accurate assessment of affective value. According to past research, increased exposure to sweet and bitter solutions has previously indicated that desensitization and habituation effects can occur. The present study aims to address these findings and wishes to determine whether innate affective responses can be influenced by specialized dieting in primates. Infant primates were used due to previous research suggesting the sensitive period in taste acquisition occurs within the first year. Results are expected to reflect past studies, such that the average number of positive facial reactions will significantly decrease when primates are exposed to sweet tasting stimuli after a period of specialized sweet dieting. Conversely, negative facial reactions are also expected to decrease when exposed to bitter tasting solution after specialized bitter dieting. Keywords: taste, habituation, desensitization, primates, affective reactions

3 Introduction As humans, we know that different types of foods seem almost naturally more pleasant or aversive than others. Certain foods only require one taste before we know that we like or dislike its flavor. This is particularly the case with sweet foods such as those with high sugar content, and with strong bitter flavors. However, such bitter tastes that may seem repulsive at first, such as wine or coffee, can become tastier with increased exposure and consumption over time, moreover they become acquired. But are these changes reflected in innate facial reactions to taste? Positive and negative affective responses have previously been observed and recorded in many species, including human and primates. These responses have been used to determine the existence of innate expressions towards sweet tastes, and innate aversive or avoidant responses to bitterness, when human infants and a variety of primates were tested (Steiner, Glaser, Hawilo, & Berridge, 2000). By quantifying the frequency and duration of facial reactions, affective expressions were shown to be significantly predictive of positive or negative valence. The study by Steiner et al. (2000) classified positive hedonic responses as rhythmic tongue protrusions, lip smacks, lip and finger sucking, and elevated mouth corners. Conversely, negative aversive reactions were indicated by gapes, eye squints, nose wrinkle, head shakes, arm flail, and frowns. The exposure to the taste appeared to be significantly powerful enough to elicit innate responses with similar intensity for both sweet and bitter tastes. There existed however a number of weaknesses and gaps in the research by Steiner et al. (2000) that this study aims to address and control against. Primarily, the diets of primates were not controlled as the study had used video recordings from a previous study that Steiner had completed and re-scored. Adult primates were also allowed to voluntarily consume tasting solutions, which may have impacted results as the responses may have been influenced by the

4 quantity of consumption. As such, it is important for this study to ensure even distribution of volume of each solution for each participant. Another important factor that Steiner et al. (2000) overlooked was the effect of development and the time for which taste preferences are acquired. Research previously done in this field of work has highlighted the effects specialization, desensitization, and the development of taste, this study is unique in that it will attempt to integrate them all together. It is therefore of interest to assess facial expressions in response to taste when diets have been controlled for beyond their natural diet. A number of studies have indicated that desensitization to oral senses can occur with increased exposure to irritating stimuli, and consequently may result in decreases of aversive responses (Cliff & Green, 1996, Karper & Bartoshuk, 1995). Yoshii and Matui (1994) also analyzed desensitization of taste after exposure to capsaicin and piperine and concluded that the taste nerves transmitting these responses are similar to the responses seen by quinine, or bitter solutions. These studies have primarily dealt with irritants such as capsaicin and menthol, without addressing primary tastes such as sweetness and bitterness alone. They have also failed to address whether only experiencing a specific taste for a prolonged period of time could influence the intensity and frequency of facial reactions. More recent studies have been focused on the habituation, acquirement, and prevalence of taste preferences. Parylak, Cottone, Sabino, Rice and Zorrilla (2012) found that rats who consumed diets containing high levels of sucrose experienced anxiety-like behaviour with the withdrawal of palatable foods, and binge eating when access was renewed. This study indicated that despite high levels of exposure to sweet foods, the level of desirability remained high. Conversely, Theunissen, Polet, Kroeze, and Schifferstein (200) found that increased exposure to sweetened yogurt resulted in taste adaptation and a perceived decrease in sweetness intensity.

5 Pepino and Mennella (2012) supported this finding in a study of obese and lean women and concluded that habituation of liking sweet tastes occurred in both samples. Despite Parylak (2012) finding that diets high in sugar concentrations remain highly palatable, this study hypothesizes that habituation will occur when foods are not withdrawn or limited. Moreover, by providing subjects with consistent exposure to primarily sweetened foods after the pre-test, they will exhibit a decrease in liking responses to sweet tastes, despite still wanting to eat them (Berridge, 2009). Habituation and adaptation to bitter and aversive oral stimuli has been supported by other studies done in the past. The main goal of this study is to further illustrate that tastes which elicit innate aversive reactions in a number of species can become increasingly pleasurable, or less aversive, with increased exposure over time. Schiffman et al. (1994) found that repeated exposure to the bitter solution quinine indeed resulted in an adaptation in subsequent taste tests. This desensitizing effect, which was also found by Yoshii and Matui (1994), indicated the potential of specialized diets to decrease aversive facial responses to quinine taste solutions. Moreover, by having a diet consist of solely bitter foods, reactions to quinine solutions are expected to significantly decrease in comparison to tests prior to specialization. The importance of specializing primates diets after they have finished breastfeeding is due to evidence that exists which indicated that taste function and neural plasticity develops postnatal. As discussed by Dominguez (2011) the development of taste is a natural process but can be influenced by experience. He found that between the ages of 3-11 months of age, human infants exposed to specific formulas will acquire that taste preference later on. These changes are seen in the central nervous system were found to be unlikely to occur in mature adults. Therefore, this period appears to be a sensitive time for taste preference acquisition. As such, it

6 will be important to wean primate infants off breastfeeding approximately 3 months after birth in order to specialize their diet. This will occur until a solid diet can be maintained independently of breastfeeding and will continue for an additional 3 months. The purpose of this study to is to determine if specialization of primate diets can alter innate taste preferences and responses to bitter and sweet solutions. According to the research there is evidence that prolonged exposure can habituate and desensitize aversive bitter tastes, as well as positive sweet tastes. It has also been found that a sensitive period exists for taste acquisition which is why it is important to study the effects during infant development. For this reason, the study by Steiner et al. (2000) needs to be extended and altered to analyze whether the innate facial responses found remain consistent when diets have been specialized immediately after breastfeeding. This may provide insight into whether or not innate human facial responses and affective value can be influenced by early specialization of diet. Based on the previous research discussed, the study proposes the following hypotheses: H1a: The frequency of pre-test and post-test positive facial responses in the natural diet condition will not significantly differ when exposed to the sweet solution. H1b: The frequency of pre-test and post-test aversive facial responses in the natural diet condition will not significantly differ when exposed to the bitter solution. Hypotheses 1 is based on the assumption that innate facial responses will follow the Steiner et al. (2000) study where infant and adult primates showed similar facial responses when no control of diet existed. Therefore it is expected that no difference in the average number of facial responses will be displayed between the pre-test and post-test.

7 H2a: The frequency of positive facial reactions in pre-test and post-test will not significantly differ in the bitter diet condition when exposed to sweet solution. H2b: The frequency of aversive facial reactions in the pre-test and post-test will significantly differ in the bitter condition when exposed to the bitter solution, such that negative aversive reactions will be significantly lower in the post-test than in the Pre-test. H3a: The frequency of pre-test and post-test positive facial reactions in the sweet diet condition will significantly differ when exposed to the sweet solution, such that positive hedonic reactions will be significantly lower in the post-test than in the pre-test. H3b: The frequency of pre-test and post-test aversive facial reactions in the sweet diet condition will not significantly differ when exposed to the bitter solution. Hypothesis 2 and 3 supports the research suggesting that aversive and positive hedonic value can become desensitized with increased exposure. No difference in sweetness or bitterness is hypothesised in their opposing condition because of the innateness of such response when increased exposure in that taste has not occurred.

Method Subjects Infant primates (n=33) will be selected from the Zoological Garden in Zurich four weeks after birth and randomly assigned to be specialized with either sweet, neutral, or bitter diets. The Zoological Garden in Zurich was selected based on its past cooperation and experience with the Steiner et al. (2000) study. Primates consist of 10 Great Apes (3 chimpanzees, 3gorilla, and 4 orangutans), 10 Old World Monkeys (4 rhesus monkeys, 3 Red-Capped Mangabey, and 3

8 Greater spot-nosed monkeys), 10 New World Monkeys (3 Golden-handed tamarins, 3 Cotton-top tamarins, 2 white-tufted-ear marmosets, and 2 Humboldts night monkeys) and 3 lemurs. Only healthy primates with no birth defects, mental or physical abnormalities will be eligible for the study. All primates will be contained in the same style cage with all the same feeding trays, and play equipment. The lighting will also be controlled to ensure that sleep schedules and activity periods are similar across species and groups. It was found that intensity of gustatory stimuli can be influenced by context and pther olfactory stimuli present (Marks, Shepard, Burger, & Chakwin, 2012; Albertella, Harris, & Boakes, 2008). Therefore testing must occur in areas which the primates are caged or exposed to regularly to ensure novel context or smells do not influence results. Primates of the same species category and diet category will be caged together to protect against isolation effects but eliminate sharing of food across diets. Apparatus Video recording will be necessary to ensure that all facial responses can be re-viewed and appropriately coded. The primary video camera will be placed in front of the primate for testing, however 2 other cameras will be placed to the sides to capture any facial reactions that occur in the case of head movements. The same concentrations of solutions will be infused into a dropper as Steiner et al. (2000) to measure facial reactions: 50 ml of 0.8 M sucrose (sweet) and 7 x 10 6 M quinine (bitter). Between each taste primates mouths will be flushed with water to control against any cross contamination and order effects.

9 Procedure All primates will be breastfeed ensure healthy post-natal development until they are able to start consuming solid food. At the point they are able to start being weaned off breast milk a pre-test will be administered, which consists of each subject being given a dropper of each solution; sweet, and bitter, with a water rinse and pause between each test. Each infant will be weaned off breastfeeding and within each species of primate they will be randomly assigned to either a sweet, bitter, or neutral diet condition (For example, within the Great Ape category, primates will be randomly assigned to each condition). By assigning primates to sweet, neutral, or bitter diets within each species it will guarantee representation of each of the specialized diets across groups. They will continue to be fed the food consistent with the specified diet for 3 months after they have discontinued breastfeeding. Primates assigned to the sweet condition will consume primarily fruit and sweetened water. Primates in the neutral diet condition will consume the natural diet fed to all other primates in the Garden; this is to establish a natural response similar to those found in the Steiner et al. (2000) study where no diet control was in place. Lastly, primates in the bitter condition will consume primarily cranberries, kale, root, dandelion, and other naturally bitter foods, as well as water containing quinine. The quantity of food given to each condition will be the same to ensure that one group or subject will not skew the results as a consequence of food deprivation or over-eating. This study will follow method procedures similar with the study done by Steiner et al. (2000). During the pre-test and the final test all subjects will be video-recorded to allow for a closer analysis of facial reactions, which will be viewed after the test has been administered. The purpose of the initial study was to determine which reactions were indicative of positive, neutral, or negative valence. Therefore, those expressions which were identified and coded as

10 significantly representative of the different levels of valence will be used as determinants of affective value. As discussed above, rhythmic tongue protrusions, lip smacks, lip and finger sucking, and elevation of mouth corners will be indicators of positive hedonic value. Neutral mouth movements will not be considered as either positive or negative valence. Gapes, eye squint, nose wrinkles, head shakes, arm flails, and frowns will be recorded as aversive valence. Both pre-test and post-test will consist of the same quantity of sweet, bitter, and neutral solutions. As discussed above, they will be administered by a 50ml dropper and each primate will be made to taste each one in each condition. This is different that Steiner et al.s (2000) study where adult primates were able to consume solutions voluntarily and therefore quantity of consumption was not controlled for. By ensuring that each participant is exposed at the same amount, accurate levels of facial expressions can more accurately be measured. Therefore, the two levels of independent variable will be specialized diet condition and taste stimuli. The dependent variable regarding frequency of facial reactions will be recorded and quantified as a measure of positive, neutral, or negative affective value and will be measured in each of the primates. The average number of racial reactions in each condition will be used for analysis. Any neutral mouth movements will be excluded from the data as they are not indicative of either positive or negative affective value. Any facial expressions that are expressed that have been found to be representative of the opposite value (ie. Positive facial reactions to bitter taste) will be quantified as negative, to show a movement in the opposite direction. Data Analysis Six paired sample t-tests will be conducted to determine whether or not significant differences exist between the average number of facial responses in the pre-test and the post-test for each sweet and bitter taste in each specialized diet condition. Each t-test will determine the

11 results of each hypothesis. The reason for no interaction or main effect analysis is that the purpose of this study is to determine the effect of specialized diet in each individual sample. A between-subjects analysis is not necessary for the purpose of this current study. Expected Results A paired-sample t-test was conducted for each of the proposed hypotheses. Pre-test and post-test means were compared for each individual condition to determine the effect of specialized dieting on sweet and bitter tastes. Hypothesis 1a addressing neutral, or unspecialized diet conditions, was supported; such that the mean number of positive facial reactions to the sweet solution in the pre-test (M = 10.59) was not significantly different (t(4) = 1.56, p = .248) than the post-test (M = 10.16). Hypothesis 1b was also supported, such that the mean number of aversive facial reactions to the bitter solution in the pre-test (M = 12.31) was not significantly different (t(4) = 2.13, p = .148) than the post-test (M = 11.44). These results support the results by Steiner et al. (2000) that even across development adult and infant primates have similar facial reactions to taste stimuli if the diets are not specialized. The two hypotheses assessing the effect of a specialized bitter diet were supported. Regarding hypothesis 2a, the mean number of positive facial reactions in the pre-test (M = 10.03) was not significantly different (t(5) = 2.97, p = .098) than the average number of post-test facial reactions (M = 11.58) when exposed to sweet solution. Hypothesis 2b was also supported, the average number of aversive facial reactions in the pre-test (M =11.52) was significantly different (t(4) = 4.13, p = .001) than the average number of aversive facial reactions in the post-test when exposed to the bitter solution (M = 5.53), such that negative aversive reactions were significantly lower in the post-test than in the pre-test.

12 Finally, the hypotheses determining the effect of a specialized sweet diet were also supported. Hypothesis 3a was supported as the mean number of positive facial reactions in the pre-test (M = 11.36) when exposed to the sweet solution was significantly different (t(4) = 2.13, p = .148) than the mean number of reactions in the post-test (M = 5.86), such that positive hedonic reactions were be significantly lower in the post-test than in the pre-test. The average number of aversive facial reactions in the pre-test (M = 11.51) and post-test (M = 12.69) in the sweet diet condition were not significantly different when exposed to the bitter solution, supporting hypothesis 3b. The data concerning hypothesis 2 and 3 support the reviewed literature that increased and consistent exposure to specific tastes can result in habituation and desensitisation to that taste. When primates were specialized with a sweet diet, a decrease in positive facial reactions was observed. This was also the case in the bitter diet condition, where a decrease in the average number of aversive facial reactions was observed.

13 Figure 1.Mean differences of facial responses to the sweet taste stimulus for each specialized diet condition. No significant difference was found for the bitter diet. The sweet diet condition did however see a significant decrease in number of positive facial responses.

14 12 10 8 6 4 2 0 Pre-Test Post-Test

Sweet Diet Neutral Diet Bitter Diet

14 Figure 2.Mean differences of facial responses in the bitter taste stimulus for each specialized diet condition. No significant difference was found for the sweet diet. The bitter diet condition did however see a significant decrease in number of negative facial responses.

14 12 10 8 6 4 2 0 Pre-Test Post-Test

Sweet Diet Neutral Diet Bitter Diet

15 References Albertella, L., Harris, J. A., & Boakes, R. A. (2008). Acquired flavour preferences: Contextual control of adaptation-level effects. The Quarterly Journal of Experimental Psychology, 61(2), 227-231. doi:10.1080/17470210701664864 Berridge, K. C. (2009). Liking and wanting food rewards: brain substrates and roles in eating disorders. Physiology & Behavior, 97, 537-550. doi: 10.1016/j.physheh.2009.02.044 Cliff, M A., & Green, B. G. (1996). Sensitization and desensitization to capsaicin and menthol in the oral cavity: interactions and individual differences. Physiology & Behavior, 59(3), 487-494. Dominguez, P. R. (2011). The study of postnatal and later development of the taste and olfactory systems using the human brain mapping approach: an update. Brain Research Bulletin. 84, 118-124.doi:10.1016/j.brainresbull.2010.12.010 Marks, L. E., Sherpard, T. G., K. Burger, & Chakwin, E. M. (2012). Flavor-intensity perception: effects of stimulus context. Physiology & Behavior, 105, 443-450. doi:10.1016/j.physbeh.2011.08.039 Parylak, S. L., Cottone, P., Sabino, V., Rice, K. C., & Zorrilla, E. P. (2012). Effects of CB1 and CRFI receptor antagonists on binge-like eating in rats with limited access to a sweet fat diet: lack of withdrawal response. Physiology & Behavior, 107(2), 231-242. Pepino, M. Y., & Mennella, J. A. (2012). Habituation to the pleasure elicited by sweetness in lean and obese women. Appetite, 58, 800-805. doi:10.1016/j.appet.2012.01.026

16 Schiffman, S. S., Pecore, S. D., Booth, B. J., Losse, M. L., Carr, B. T., Sattely-Miller, E., Graham, B. G., & Warwick, Z.S. (1994). Adaptations of sweeteners in water and in tannic acid solutions. Physiology & Behavior, 55(3), 547-559. Steiner, J. E., Glaser, D., Hawilo, M. E., & Berridge, K. C. (2000). Comparative expression of hedonic impact: affective reactions to taste by human infants and other primates. Theunissen, M. J. M., Potel, I. A., Kroeze, J. H. A., & Schifferstein, H. N.J. (2000). Taste adaptation during the eating of sweetened yogurt. Appetite, 34, 21-27. doi:10.1006/appe.1999.0275 Yoshii, K., & Matui, T. (1994). Taste responses of bullfrog to pungent stimuli. Brain Research, 637, 68-72.