Applied Animal Behaviour Science 181 (2016) 129136

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Applied Animal Behaviour Science

journal homepage: www.elsevier.com/locate/applanim

Facial expressions and other behavioral responses to pleasant and

unpleasant tastes in cats (Felis silvestris catus)
Michaela Hanson a , Susan M. Jojola b , Nancy E. Rawson b , Melissa Crowe b ,
Matthias Laska a,
a
b

IFM Biology, Linkping University, Campus Valla, 581 83 Linkping, Sweden

AFB International, 3 Research Park Drive, St. Charles, MO 63304, USA

a r t i c l e

i n f o

Article history:
Received 24 December 2015
Received in revised form 26 May 2016
Accepted 27 May 2016
Available online 4 June 2016
Keywords:
Behavior
Cat
Felis silvestris catus
Taste reactivity
l-Proline
Quinine monohydrochloride

a b s t r a c t
The goal of the present study was to assess how cats react to tastes previously reported to be preferred or avoided relative to water. To this end, the facial and behavioral reactions of 13 cats to different
concentrations of l-Proline and quinine monohydrochloride (QHCl) as well as mixtures with different
concentrations of the two substances were assessed using a two-bottle preference test of short duration.
The cats were videotaped and the frequency and duration of different behaviors were analyzed. Signicant differences in the cats behavior in response to the taste quality of the different solutions included,
but were not limited to, Tongue Protrusions (p < 0.039), Mouth smacks (p = 0.008) and Nose Licks (p = 0.011)
with four different stimulus concentrations. The cats responded to preferred taste by keeping their Eyes
half-closed (p = 0.017) for signicantly longer periods of time with four different stimulus concentrations
compared to a water control. When encountering mixtures containing l-Proline and QHCl the cats performed Tongue protrusion gapes (p < 0.038) signicantly more frequently with three different stimulus
concentrations compared to an l-Proline control. A stepwise increase in the concentration of l-Proline
from 5 mM to 500 mM in mixtures with 50 M QHCl did not overcome the negative impact of the bitter
taste on intake. The results of the present study suggest that behavioral responses provide an additional
dimension and may be more informative than consumption data alone to assess whether cats perceive
tastes as pleasant or unpleasant. Thus, the analysis of behavioral responses to different taste qualities
may be a useful tool to assess and improve the acceptance of commercial food by cats.
2016 Elsevier B.V. All rights reserved.

1. Introduction
To develop palatable food for cats we have to understand what
is palatable to a cat and how to evaluate a cats experience with
a food item. Palatability is often measured by consumption, and
the taste experience can be inferred by observable expressions,
or taste reactivity. An important aspect to consider in the evaluation of what is palatable to cats is that their behaviors and senses
have evolved differently than those of humans and other species
(e.g., Bradshaw et al., 1996; Bradshaw, 2006). For example, taste
buds of the domestic cat are highly responsive to amino acids
whereas responses to mono- and disaccharides (sugars) are almost
non-existent (Bradshaw et al., 1996). Cats have been reported to
prefer the taste of l-Proline (White and Boudreau, 1975; Bradshaw,
1991) This amino acid is described by humans as having a complex

Corresponding author at: IFM Biology, Linkping University, SE-581 83

Linkping, Sweden.
E-mail address: malas@ifm.liu.se (M. Laska).
http://dx.doi.org/10.1016/j.applanim.2016.05.031
0168-1591/ 2016 Elsevier B.V. All rights reserved.

sweet taste with components of both salty and sour (Schiffman and
Sennewald, 1981). Cats are unable to detect the taste of simple saccharides, and do not display behavioral responses to sugars such as
sucrose (Li et al., 2006; Bradshaw et al., 1996). Tastes described as
bitter by humans are usually avoided by many mammals, including
cats. Bitter tastes can sometimes occur in commercial cat food due
to the addition of important nutrients (Zaghini and Biagi, 2005).
Taste buds of cats respond to quinine, phenylthiocarbamide, aloin
and denatonium which all have a distinct bitter taste (Cohen et al.,
1955; Sandau et al., 2015). Quinine monohydrochloride (QHCl) is
also described by humans as having a distinctly bitter taste and is
commonly used when investigating responses to bitter taste in both
human subjects and non-human mammals (Steiner and Glaser,
1984; Berridge, 2000; Jankunis and Whishaw, 2013).
Behavioral responses to different tastes, known as taste reactivity, have been reported in a variety of mammals including
humans, non-human primates, rats, rabbits, hamsters, horses, and
cats (Ganchrow et al., 1979; Ganchrow et al., 1986; Brining et al.,
2003; Steiner and Glaser, 1995; Bartlett et al., 1999; Berridge, 2000;
Van den Bos et al., 2000; Steiner et al., 2001; Ueno et al., 2004;

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M. Hanson et al. / Applied Animal Behaviour Science 181 (2016) 129136

Jankunis and Whishaw, 2013). When experiencing tastes regarded

as pleasant by the animal, facial expressions that involve smacking
with the lips, tongue protrusion and relaxation of the facial muscles
are commonly observed. Tastes perceived as unpleasant, in contrast, elicit facial expressions that commonly involve gaping mouth
movements, head shakes and ailing with the limbs. The benet of
taking such behaviors related to the taste experience into consideration, as opposed to only looking at consumption data, is that
we get a more precise idea of how the animal experiences a taste
and gain better insight into the food selection process (Grill and
Norgren, 1978).
The goal of the present study was therefore to assess how cats
react to pleasant or unpleasant taste solutions with regard to consumption and behavior, how they react to taste mixtures, and if
it was possible to mask the unpleasant taste with a pleasant one.
We used l-Proline as the stimulus to elicit behavioral responses
indicative of a pleasant taste experience. QHCl was used as the stimulus to elicit behavioral responses indicative of an unpleasant taste
experience.
2. Methods
2.1. Animals
Thirteen adult cats (Felis silvestris catus), 7 males and 6 females,
of the breed Domestic Short Hair (DSH) were used in the study.
The cats were bred for research purposes at Liberty Research, Inc.
(Waverly, NY, USA) and housed at AFB Internationals Palatability
Assessment Resource Center (PARC) where the study took place
from July to December 2014. All cats were around 2.5 years old
when the experiments were initiated to ensure that they had fully
developed their senses related to the experience of taste as well
as their preference for certain tastes (Van den Bos et al., 2000;
Bradshaw, 2006). Both males and females were neutered at the age
of 4.5 months to eliminate hormonal factors such as the oestrous
cycle that could have an impact on the taste sensation (Van den Bos
et al., 2000). Furthermore, the cats were healthy individuals without any history of allergies to the taste stimuli used in the tests and
had no history of serious injuries or behavioral problems. The cats
were free from upper respiratory tract infections (URI) and were
regularly monitored by animal technicians. The cats were housed
in groups of 24 individuals, 12 males and 12 females, and were
allowed to move freely together in a 5 5 m large room. The cats
had ad libitum access to water, litter boxes, toys and during daytime a 3 5 m large outdoor porch. The experiments reported here
were all performed at AFB International which is a registered class
R research facility under the Animal Welfare Act certicate number 43-R-0119 and comply with current U.S. laws. This study was
approved by the Sr. Director of Product Development and Palatability of AFB International. Annual inspections of the facility are
performed by the U.S. Department of Agriculture.
2.2. Experimental set-up
Two 17 ml metal spouts were mounted to a Plexiglass door of a
75 61 72 cm metal cage as the set-up for two-bottle preference
tests of short duration. Solutions (14 ml) were added to the clean
spouts prior to each trial. Vinyl gloves were worn at all times when
prepping the spouts and handling solutions to prevent contamination with human odor. A digital lick-o-meter counter (model 86063,
Lafayette Instrument Company, Lafayette, IN, USA) was mounted to
each spout and registered the number of times the cat came into
direct contact with the spout.
Observations took place from 09:30 to 14:30, four days a week.
Daily feedings ended at 06:00 at which time food was removed,

and no food was accessible to the cats during trials. Specic cats
were always tested either in the morning or afternoon to control
for potential differences in performance due to variation in time of
day. Each cat participated in only one 5 min trial per day and one
cat was tested at a time.
l-Proline (CAS# 147-85-3) and quinine monohydrochloride
dihydrate (QHCl) (CAS# 6119-47-7) were used throughout the
study to assess behavioral responses based on their respective taste
qualities. The taste solutions and the water used as control stimulus were stored in glass jugs sealed with a lid and kept in a fridge at
6 C. The solutions and the water were removed from fridge storage
2 h prior to the start of testing in order to have the solutions closer
to room temperature. The pH of the taste solutions was checked
when making the stock solutions from which the taste solutions
were derived.
2.3. Video data and coding procedure
We video-recorded trials using a Sony (Tokyo, Japan) HDRCX380 camera on a tripod placed 2.3 m in front of the cage. Videos
were later coded using The Observer XT software v11.5 (Noldus;
Wageningen, The Netherlands) according to a coding scheme of
dened behaviors. Behaviors and facial expressions included in
the study (Table 1.; for a more detailed description of analyzed
behaviors, see supplementary material) were based on the P02
Feline Behavior Coding Scheme for Liquid Stimuli from AFB International and reports of commonly occurring facial expressions and
behaviors in mammals other than cats when experiencing tastes
perceived as pleasant or unpleasant (Van den Bos et al., 2000;
Berridge, 2000; Steiner et al., 2001; Jankunis and Whishaw, 2013).
Behaviors and facial expressions in the study were divided into
state behaviors (e.g. snifng) which were measured in their duration and frequency, and behavioral events (e.g. mouth movements)
which were measured in their frequency. For a trial to be regarded
as successful, the cat had to interact with at least one of the spouts in
the cage by coming within a 2.5 cm radius from the spout and sniff
at the spout. Trials in which the cat failed to do so were eliminated
from the data set. Coders were blind to the treatment assignments
to prevent bias. Reliability analyses were run to obtain agreement
scores between and within coders as a measure of their precision
and consistency in identifying the occurrence of relevant behaviors.
In order to test for reliability, coder 1 observed 3 videos twice and
coder 2 observed 2 videos twice. Both coders had previous working experience with cats. One of the coders had previously worked
with the cats used in the study. When running reliability analysis, the same videos were used for both coders in order to check
for inter-observer agreement. Reliability analysis was done from
these double viewings from which an average reliability score was
calculated. The reliability score was calculated by index of concordance. Index of concordance was calculated by dividing the number
of agreements with the combined number of agreements and disagreements. The scores range from 0 to 1, with 1 indicating perfect
agreement. Coder 1 had an intra-observer score of 0.98 for duration and 0.89 for frequency and coder 2 had a score of 0.94 for
duration and 0.71 for frequency. The inter-observer score was 0.93
for duration and 0.61 for frequency.
2.4. Experiments 1 & 2
Cats were presented with either l-Proline (Experiment 1) or
QHCl (Experiment 2) in one of the spouts and a control of distilled water in the other spout. Five different concentrations of
l-Proline and QHCl, respectively, were tested. The lowest concentration, 0.05 mM for l-Proline and 0.005 M for QHCl, was
presented rst and increased in 10-fold concentration steps with
each week of testing. The strongest concentration presented to the

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131

Table 1
Behaviors analyzed throughout the study.
Behavior
State Behaviors
Not drinking or snifng
Drinking from the spout
Drinking dripped solution
Snifng at the spouts
Snifng the interior of the cage
Eyes open/wide-open
Eyes half closed
Eyes closed
Ears pointed upward and/or forward
Ears lowered and/or outward
Ears in different positions
Behavioral events
Licks nose
Licking whiskers
Tongue protrusions
Tongue protrusion gapes
Mouth smacks
Miscellaneous

Description

Eyes were only analyzed while the cat was interacting

with the solutions
Ears were only analyzed while the cat was interacting with
the solutions

Flicks tongue quickly over the nose

Cat sweeps its tongue to the side of it face
Tongue directed forward out of the mouth
Tongue directed forward out of the mouth while gaping
Single chewing motion while slightly opening the mouth

State behaviors were measured both in frequency and duration while behavioral events were only measured in frequency.

cats was 500 mM for l-Proline and 50 M for QHCl. Thus the ve
concentrations for l-Proline were 0.05, 0.5, 5, 50 and 500 mM. For
experiments with QHCl, the ve concentrations were 0.005, 0.05,
0.5, 5 and 50 M. The order of stimulus presentation (from low to
high concentrations) was chosen to avoid affecting the cats sensitivity to the different compounds. A previous study found that the
preference of cats for l-Proline peaks at a concentration of 50 mM
(White and Boudreau, 1975). Thus, we bracketed this concentration
in our study. Carpenter (1956) found that cats responded to QHCl
at concentrations as low as 5 M. We chose to include that level
and one higher concentration so that possible aversive responses
towards the solution were likely to occur. Each cat was presented
with one concentration of l-Proline and QHCl, respectively, twice
over four days. Solutions were presented alternately to provide a
different stimulus each day and to encourage the cats to continue
investigating the spouts.
2.5. Experiment 3
Cats were presented with a mixture of l-Proline and QHCl in
one spout and a 50 mM l-Proline as the alternative stimulus in the
other spout. This experiment investigated the masking potential
of l-Proline and the cats behavioral responses based on the taste
qualities of a mixture of two different compounds versus a single
compound solution. The concentration of l-Proline was systematically increased with each mixture while the concentration of QHCl
remained constant in all of the mixtures. Ranges of preference and
avoidance in cats for l-Proline reported by White and Boudreau
(1975) and for QHCl reported by Carpenter (1956), as well as
indications of the range of preference and avoidance for the solutions in the current study, were used to derive concentrations for
mixtures. Similar to Experiments 1 & 2, mixtures of l-Proline and
QHCl started at the lowest concentration and increased each week.
The concentrations of l-Proline in the mixtures were 5, 25, 50, 250
and 500 mM, respectively, while the concentration of QHCl was
kept at 50 M in all the mixtures. Each stimulus combination was
presented twice to the cats over a time period of two days. The cats
were, thus, presented two different stimulus combinations each
week.
2.6. Data analysis
2.6.1. Consumption
The number of licks, as registered by lickometer counters, was
compared for the treatment and alternative stimulus, and com-

pared between concentration levels. To avoid pseudoreplication,

the two exposures to the same solution and concentration were
pooled for each cat. We used Wilcoxons test for pairwise observations for within-series analyses and the Mann-Whitney U test for
between-series analyses. We used a Friedmans test (ANOVA) for
comparisons between more than two data sets, e.g. for comparing between the ve different concentrations tested with a given
taste stimulus. When Friedmans test yielded a signicant result,
this was followed by pairwise comparisons using the Wilcoxons
test in order to assess which data sets were responsible. Alpha was
set at 0.05.
All analyses were run in SPSS v. 22 IBM SPSS Statistics. Degrees
of freedom for all tests were calculated to 12.

2.6.2. Taste reactivity

Behavioral data from the coded videos were obtained by running the Behavioral Analysis option in the Observer XT software
v11.5 (Noldus; Wageningen, The Netherlands). The Behavioral
Analysis helped us to answer the following questions focusing on
three different scenarios: What behaviors occurred as the cat in
any way sampled the solution?, What other behaviors occurred
as the cat performed the behavior drinking? and nally What
behaviors occurred during the individual cats test session as a
whole?.
With regard to the rst question, we looked at what behaviors
occurred both when the cat was snifng the spout and sampling
the solution by direct ingestion. With regard to the third question,
observed behaviors during the full test session were ascribed to the
solution (treatment or alternative stimulus) located on the same
side (left or right) that the behavior occurred.
To account for variances in time spent at each stimulus due to the
different characteristics of the stimuli, duration of a specic behavior exhibited at the experimental stimulus was divided by the total
duration the cat exhibited that behavior when interacting with both
the experimental stimulus and the control stimulus. For example,
the time spent with their Eyes open/wide-open at the l-Proline spout
was divided by the total time spent with their Eyes open/wide-open
at both the l-Proline and water spouts. This procedure normalized
data by describing a ratio of time that a behavior was exhibited
at a specic treatment relative to the overall time that behavior
was exhibited by the cat. Data presented are not normalized unless
specied.

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Fig. 2. A cat having its eyes half closed while sampling a solution.
Fig. 1. Registered number of licks in Experiment 1 (a) and Experiment 2 (b). Mean
values (SE) of the total number of registered licks at spouts containing different
concentrations of solutions (black squares) or water (white circles), respectively,
are given. An asterisk indicates a signicant difference between solution and water
control at the corresponding concentration (p < 0.05).

3. Results
3.1. Experiment 1
Cats licked l-Proline signicantly more often than water in
trials with 50 mM (Wilcoxon, Z = 2.395, p = 0.017) and 500 mM
(Wilcoxon, Z = 2.202, p = 0.028) (Fig. 1a). A peak in the number of licks was observed at 50 mM l-Proline. Cats licked at the
spouts containing water to a similar extent with all ve concentrations of l-proline. When comparing the total number of
licks between the different concentrations of l-Proline in Experiment 1, a signicant difference was found between 0.05 and
500 mM (Wilcoxon, Z = 2.040, p = 0.041) as well as between 0.5
and 500 mM (Wilcoxon, Z = 2.411, p = 0.016). No signicant difference in the total number of licks between the water tested together
with the different concentrations of l-Proline was found (Friedman,
p = 0.479).
Signicant differences in behaviors were only observed in trials with 50 and 500 mM of l-Proline versus water (Fig. 3), but
not in trials with the lower concentrations tested. While sampling l-Proline at 50 mM, the cats had their Eyes open/wide-open
signicantly more frequently compared to when the cats were sampling water (Wilcoxon, Z = 1.960, p = 0.050). The cats had their
Eyes open/wide-open for signicantly longer periods of time when
sampling l-Proline at 50 and 500 mM compared to when sampling l-Proline at 5 mM (Wilcoxon, Z = 2.666, p = 0.008; Z = 2.666,
p = 0.008) The duration of the cats having their Eyes open/wideopen (Wilcoxon, Z = 1.992, p = 0.46), half closed (Fig. 2) (Wilcoxon,
Z = 2.394, p = 0.017) or closed (Wilcoxon, Z = 2.201, p = 0.028,)
were all signicantly longer when sampling from spouts containing l-Proline compared to when sampling from water spouts.
Time spent sampling from 50 mM and 500 mM l-Proline was
signicantly longer than time spent sampling water (Wilcoxon,
Z = 2.201, p = 0.28; Z = 2.621, p = 0.009), to account for this the

data were normalized. After normalizing the values to account

for disproportionate time at each stimulus, Eyes half closed still
occurred signicantly longer when sampling l-Proline at 50 mM
compared to water (Wilcoxon, Z = 2.197, p = 0.028). At 500 mM,
the cats had their Eyes open/wide-open (Wilcoxon, Z = 2.481,
p = 0.013) and half closed (Wilcoxon, Z = 2.201, p = 0.028) for a
signicantly longer period of time when sampling l-Proline compared to when sampling water when analyzing non-normalized
data. The normalized data for 500 mM l-Proline, however, indicated no signicant difference in the aforementioned behaviors
between treatment and water. Both when sampling 50 and 500 mM
l-Proline, the cats positioned their Ears upward and/or forward for
signicantly longer periods of time compared to when sampling
water (Wilcoxon, Z = 2.201, p = 0.028; Z = 2.341, p = 0.019)
The frequency with which the cats had their Eyes open/wideopen (Wilcoxon, Z = 1.960, p = 0.050,) or had their Eyes closed
(Wilcoxon, Z = 2.214, p = 0.027) was signicantly higher when
sampling 50 mM l-Proline compared to water. The frequency of
the cats having their Eyes closed was also signicantly higher
when the cats were sampling 500 mM l-Proline compared to
water (Wilcoxon, Z = 2.132, p = 0.033). During sampling of 50 mM
l-Proline the cats performed Tongue protrusion behavior significantly more frequently than with water (Wilcoxon, Z = 2.060,
p = 0.039). When sampling from 500 mM l-Proline the cats performed Tongue protrusions (Wilcoxon, Z = 2.519, p = 0.012), Mouth
smacks (Wilcoxon, Z = 2.641, p = 0.008) and Nose licks (Wilcoxon,
Z = 2.530, p = 0.011) signicantly more often than when sampling water. The cats performed Tongue Protrusions signicantly
more frequently when sampling 500 mM l-Proline compared
to when sampling l-Proline at 0.05, 5 or 50 mM (Wilcoxon,
Z = 2.439, p = 0.015; Z = 2.145, p = 0.032; Z = 2.047, p = 0.041,).
Mouth Smacks occurred signicantly more often when the cats
were sampling 500 mM l-Proline compared to when sampling
l-Proline at 5 mM (Wilcoxon, Z = 2.263, p = 0.024). Time spent
Snifng at the spouts was signicantly longer at spouts containing
50 or 500 mM l-Proline compared to water (Wilcoxon, Z = 2.132,
p = 0.033; Z = 1.992 p = 0.046).
Time spent drinking 500 mM l-Proline was signicantly longer
than time spent drinking water (Wilcoxon, Z = 2.510, p = 0.012).

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133

Fig. 3. Comparison between treatment and reference solution where signicant differences were found in duration (a) and/or frequency (b) of behaviors concerning the eyes
and ears. Black bars represent the treatment specied on the x-axis. White bars represent the corresponding reference solution. Mean values (SE) of the total duration (a)
and total number (Fig. 3b) of observed behaviors concerning the eyes and ears are given.

When drinking 500 mM l-Proline, the cats had their Eyes half closed
(Wilcoxon, Z = 2.073, p = 0.038) for a signicantly longer period of
time compared to when drinking water. The cats also positioned
their Ears upward and/or forward for signicantly longer periods of
time (Wilcoxon, Z = 2.118, p = 0.034) as well as more frequently
(Wilcoxon, Z = 1.984, p = 0.047) while drinking the 500 mM lProline compared to the water.
3.2. Experiment 2
Cats did not lick spouts containing QHCl signicantly less often
than they did spouts containing water at any of the tested concentrations (Wilcoxon, p > 0.05) (Fig. 1b). When comparing the total
number of licks between the different concentrations of QHCl in
Experiment 2, no signicant difference in the total number of licks
was found (Friedman, p = 0.208). Similarly, there was no signicant
difference in the total number of licks between the water tested
together with the different concentrations of QHCl (Friedman,
p = 0.425). Despite the lack of a signicant difference in consumption of QHCl, signicant differences in behavior were observed
(Fig. 3). Most notably, while sampling from the water, Tongue protrusions (indicative of a pleasant taste experience) were observed
signicantly more often compared with when cats were sampling
from the 50 M QHCl (Wilcoxon, Z = 2.156, p = 0.031). Tongue

Protrusions occurred signicantly more often with water tested

together with QHCl at 50 M than water tested together with
QHCl at 0.005 or 5 M (Wilcoxon, Z = 2.214, p = 0.027; Z = 2.041,
p = 0.041).
Further, the cats had their Eyes closed signicantly more frequently when sampling 0.005 M QHCl compared with water
(Wilcoxon, Z = 2.070, p = 0.038). When sampling 5 M QHCl, the
duration with which the cats held their Ears lowered and/or outward was signicantly shorter (Wilcoxon, Z = 2.023, p = 0.043) as
well as signicantly less frequent (Wilcoxon, Z = -2.070, p = 0.038)
compared to when sampling water. When drinking 50 M QHCl the
cats had their Ears upward and/or forward for signicantly shorter
periods of time (Wilcoxon, Z = 2.293, p = 0.022). Instead, the cats
held their Ears in different position for signicantly longer periods
of time when drinking 50 M QHCl compared to water (Wilcoxon,
Z = 1.992, p = 0.046).
3.3. Experiment 3
The cats licked spouts containing 50 mM l-Proline signicantly more often compared to mixtures of 50 M QHCl + l-Proline
at 5 (Wilcoxon, Z = 2.358, p = 0.018), 25 (Wilcoxon, Z = 2.490,
p = 0.013), 50 (Wilcoxon, Z = 2.132, p = 0.033) and 250 mM
(Wilcoxon, Z = 2.412, p = 0.016) (Fig. 4). In trials of mixtures with

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M. Hanson et al. / Applied Animal Behaviour Science 181 (2016) 129136

Fig. 4. Registered number of licks in Experiment 3. Mean values (SE) of the total
number of registered licks at spouts containing different concentrations of the lProline and QHCl mixture (black squares) or 50 mM l-Proline control (white circles),
respectively, are given. An asterisk indicates a signicant difference between the
l-Proline and QHCl mixture and the l-Proline control at the corresponding concentration (p < 0.05).

500 mM l-Proline + 50 M QHCl, the cats did not signicantly differ

in the number of licks at spouts containing the mixture compared to spouts containing the 50 mM l-Proline alone (Wilcoxon,
p > 0.05). When comparing the total number of licks between the
different concentrations of the l-Proline and QHCl mixtures in
Experiment 3, we found that the 5 mM l-Proline + 50 M QHCl mixture received signicantly fewer licks than the 50, 250 and 500 mM
l-Proline + 50 M QHCl mixtures (Wilcoxon, Z = 2.046, p = 0.41;
Z = 2.703, p = 0.007; Z = 2.984, p = 0.003). No signicant difference in total number of licks between the l-Proline tested together
with the different concentrations of the l-Proline + QHCl mixtures
was found (Friedman, p = 0.546).
When sampling a 5 mM l-Proline + 50 M QHCl mixture the cats
held their Eyes open/wide- open (Wilcoxon, Z = 2.045, p = 0.041) or
half closed (Wilcoxon, Z = 2.497, p = 0.013) for signicantly shorter
periods of time compared to the 50 mM l-Proline (Fig. 3). The time
spent sampling from the 50 mM l-Proline was signicantly longer
than the time spent sampling the 5 mM l-Proline + 50 M QHCl
mixture (Wilcoxon, Z = 2.845, p = 0.004), to account for this the
data were normalized. When normalizing the data, it was conrmed that the cats spent a signicantly shorter duration of their
time with their eyes either open/wide-open (Wilcoxon, Z = 2.191,
p = 0.028) or half closed (Wilcoxon, Z = 2.395, p = 0.017) when sampling the 5 mM l-Proline + 50 M QHCl mixture compared to the
50 mM l-Proline. Just as in trials with the 5 mM l-Proline + 50 M
QHCl mixture, the duration of the cats having their Eyes half closed
was signicantly longer during sampling from the 50 mM l-Proline
compared to when the cats were sampling from the 250 mM lProline + 50 M QHCl mixture when considering non-normalized
data (Wilcoxon, Z = 2.132, p = 0.033). The duration of Eyes half
closed was signicantly shorter with the 5 mM l-Proline + 50 M
QHCl mixture than with the 250 (Wilcoxon, Z = 2756, p = 0.006)
and 500 mM l-Proline + 50 M QHCl mixture (Wilcoxon, Z = 2984,
p = 0.003) The duration with which the cats held their Ears upand/or forward was signicantly shorter when sampling the 5 mM
l-Proline + 50 M QHCl mixture compared to the 50 mM l-Proline
(Wilcoxon, Z = 2.802, p = 0.005).
With mixtures containing 5 or 250 mM l-Proline + 50 M QHCl
the time the cats spent drinking from the spout was signicantly
shorter compared to the 50 mM l-Proline (Wilcoxon, Z = 2.310,
p = 0.021; Z = 2.201, p = 0.028). When drinking from a 5 mM lProline + 50 M QHCl mixture the cats held their Eyes half closed
(Wilcoxon, Z = 2192, p = 0.028) for signicantly shorter periods of
time compared to the 50 mM l-Proline. The cats also had their Ears
positioned up- and/or forward (Wilcoxon, Z = -2073, p = 0.038) for
signicantly shorter periods of time when drinking from the 5 mM
l-Proline + 50 M QHCl mixture compared to the 50 mM l-Proline.

Fig. 5. A cat performing a tongue protrusion gape, sticking its tongue out and at the
same time gaping with its mouth.

Tongue protrusion gapes (Fig. 5) were observed signicantly

more often when the cats were near the 250 mM l-Proline + 50 M
QHCl mixture compared to the 50 mM l-Proline (Wilcoxon,
Z = 2.121, p = 0.034). An increase in Tongue protrusion gapes was
suggested in Experiment 2 but was not signicant, which may be
related to the nding that the cats licked signicantly more at the
50 M QHCl + 250 mM l-Proline mixture than at the 50 M QHCl
solution in Experiment 2 (Mann-Whitney U, Z = 2.031, p = 0.044).
4. Discussion
Our rst goal was to assess how cats react to pleasant tastes.
The cats performed behaviors such as Snifng at the spouts, Eyes
half closed, Tongue protrusions, Nose licks and Mouth smacks significantly more often with certain concentrations of l-proline than
with water. With regard to differences in consumption, the cats displayed a signicant preference for l-Proline at 50 and 500 mM over
water. Our second goal was to assess how cats react to unpleasant
tastes. Signicant differences in the behavior of the cats with QHCl
and water were observed with Tongue protrusions being more frequent when the cats were sampling water. Based on consumption,
no aversion or preference for QHCl compared to water was found.
Our third goal was to assess how cats react to mixtures of pleasant and unpleasant tastes and possible masking effects. Here, the
cats displayed a preference for solutions of l-Proline without the
addition of QHCl. The only concentration where licking was similar between the mixture and the l-Proline alone was at the highest
concentration of l-Proline. However, this was likely not a masking effect for l-Proline as the difference was due to a decrease in
licking for the l-Proline rather than an increase for the mixture.
Consistent with the results in Experiment 2, the cats differed signicantly between the l-Proline + QHCl mixtures and the l-Proline
alone in behaviors such as Eyes half closed and Tongue protrusion
gapes.
We identied specic behaviors that can be attributed to a pleasant taste experience for the cat. In both Experiment 1 and Experiment
3, the cats responded to preferred concentrations of l-Proline by
having their Eyes half closed for signicantly longer periods of time
compared to the alternative stimulus. Non-human primates have

M. Hanson et al. / Applied Animal Behaviour Science 181 (2016) 129136

been reported to show a similar response to tastes perceived as

pleasant by having their eyes slightly, but not fully, closed in a
relaxed manner (Steiner and Glaser, 1984). A commonly reported
response to pleasant tastes in newborn humans, non-human primates and rabbits is that the animal tends to have a relaxed facial
expression (Steiner, 1973; Ganchrow et al., 1979, 1983; Steiner
and Glaser, 1984). Studies in cattle have shown that having the
eyes slightly, but not fully closed, is possibly a sign of general contentedness which one should keep in mind when considering the
implications of the behavior (Sandem et al., 2002). Based on our
preference data and previous studies, that the cats having their
Eyes half closed may represent a form of relaxed facial expression
in this species. This indicates that cats, just as the previously mentioned mammals, respond to pleasant tastes with a relaxed facial
expression. Another possibility is that holding the Eyes half closed is
a behavioral response to pleasant tastes specic to cats. The fact that
Eyes half closed while sampling l-Proline occurred in both Experiment 1 and Experiment 3 further underlines the signicance of this
behavior as a part of the cats behavioral repertoire to pleasant
tastes. It is important to mention that the behavior Eyes half closed
should not be confused with the eye squinch often seen as a reaction to unpleasant taste in human infants (Steiner, 1973 via Steiner
et al., 2001). The eye squinch refers to the eyes being shut and a visible compression of the periorbital muscles which is not consistent
with a relaxed facial expression.
Tongue protrusions are one of the most commonly described
reactions to tastes perceived as pleasant and have been reported
to occur in humans, non-human primates, rats, mice, rabbits and
horses (Grill and Norgren, 1978; Ganchrow et al., 1979; Steiner and
Glaser, 1995; Kiefer et al., 1998; Steiner et al., 2001; Ueno et al.,
2004; Jankunis and Whishaw, 2013). The results of the present
study show that cats are no exception to this. Tongue protrusions
were signicantly more frequent when sampling from spouts containing preferred concentrations of l-Proline compared to water
or mixtures of l-Proline + QHCl. The present ndings indicate that
Tongue protrusions are a behavioral response to tastes perceived
as pleasant by cats, similarly to what has been observed in other
mammalian species.
When sampling l-Proline at 500 mM, Mouth smacks and Nose
licks became signicantly more frequent compared to the lower
concentrations of l-Proline used in the present study. In great apes
and gibbons and newborn humans the behavior lip smack, similar
to the behavior mouth smack, is indicative of a pleasant taste experience (Steiner and Glaser, 1995). Rats have also been reported to
perform mouth smacks as a response to pleasant taste (Grill and
Norgren, 1978). The behavior Nose lick occurred signicantly more
often when the cats were sampling l-Proline at 500 mM. Steiner
et al. (2001) reported that a behavior dened as an upwards tongue
protrusion in New World monkeys, similar to the nose lick behavior in cats, was indicative of a pleasant taste experience.
The cats were snifng at high concentrations of l-Proline for
signicantly longer durations than they were snifng at water or
l-Proline and QHCl mixtures. This suggests that the cats were able
to detect the odor of the amino acid. l-Proline is known to have
a detectable odor for humans, spider monkeys, and mice, when
presented at mM concentrations (Laska, 2010; Walln et al., 2012).
We found a negative impact of QHCl on the taste experience
for cats during tests with mixtures of l-Proline + QHCl. The changes
that occurred both with regard to behavior as well as the registered number of licks show that QHCl had negative effects on how
cats experienced the mixture. Carpenter (1956) reported that cats
actively avoided consuming QHCl at 5 M, which was not observed
in the present study. One possible explanation for the difference
between the results is that Carpenters cats were exposed to QHCl
for 48 h while in the present study the cats only had access to QHCl
for 5 min. It is possible that with a 48 h exposure the cats behavior

135

was affected by postingestive factors so that variables other than

taste alone may have inuenced the aversion towards QHCl in cats
as reported by Carpenter.
Even though no aversion to QHCl was found in Experiment 2, it
is interesting to note the signicant differences in behavior occurring during these trials. Tongue protrusions that are often connected
to a pleasant taste experience were observed to occur more often
while the cats were sampling water when paired with QHCl. However, Ganchrow et al. (1983) stated that water evokes the least
intense facial expressions in terms of behavioral response to taste
in newborn humans. Rather than an increase in tongue protrusions while sampling water it is more likely that there was a lower
frequency of Tongue protrusions while sampling QHCl. One of the
more commonly reported reactions to bitter taste are different
varieties of the Tongue protrusion gape where the animal opens
its mouth and then sticks its tongue out. This behavior has been
observed in non-human primates, rats, rabbits, mice and horses
(Grill and Norgren, 1978; Ganchrow et al., 1979; Steiner and Glaser,
1984; Kiefer et al., 1998; Ueno et al., 2004; Jankunis and Whishaw,
2013). The cats in the present study were observed to perform the
behavior Tongue protrusion gape when near a spout containing an
l-Proline + QHCl mixture. The Tongue protrusion gape behavior was
consistently observed to occur either during drinking or within 20 s
after nishing drinking the l-Proline + QHCl mixture. Thus, Tongue
protrusion gapes always occurred after a cat had tasted the solution.
An increase in Tongue protrusion gapes was observed in Experiment
2 as well but the difference did not reach statistical signicance.
Most likely, the cats were more exposed to the possible side taste of
QHCl in Experiment 3 as they licked at the QHCl + l-Proline mixture
to a signicantly greater extent than they licked at the QHCl solution in Experiment 2. This would explain Tongue protrusion gapes
being more frequent in Experiment 3 as a behavioral reaction to
the unpleasant taste the behavior may require a more intense or
extended exposure to the taste to be evoked. Behaviors related to
positioning of the ears have been connected to an unpleasant taste
experience in horses (Jankunis and Whishaw, 2013). In the present
study, signicant differences in how the cats positioned their ears
while interacting with solutions of QHCl were observed but the way
the cats positioned them was inconsistent and no systematic connection between the taste quality of QHCl and positioning of the
ears was found.

5. Conclusion
One of the aims for the present study was to expand upon
the current knowledge on behavioral reactions of cats to taste as
a complementary parameter for future studies when evaluating
palatability of food. The cats responded to tastes regarded as pleasant by having their Eyes half closed for signicantly longer periods
of time compared to the water. They also performed Tongue protrusions, Mouth smacks and Nose licks signicantly more frequently as
a reaction to tastes regarded as pleasant compared to water. During
tests with mixtures of l-Proline + QHCl, the behavior Tongue protrusion gape occurred signicantly more often after a cat had tasted the
mixture compared to l-Proline presented as the alternative stimulus. The present study suggests that behavioral responses to tastes
perceived as pleasant or unpleasant can be utilized in future studies on how cats experience different tastes. Knowledge on how cats
react to pleasant and unpleasant tastes gives us a more nuanced
picture on how a food item is perceived compared to only relying on consumption data. This, in turn, may help us to identify
more precisely what a cat nds pleasant in terms of different taste
components, and to identify possible side tastes in commercial cat
food.

136

M. Hanson et al. / Applied Animal Behaviour Science 181 (2016) 129136

Acknowledgements
We thank Annetta Duggan and Joya Johnson for their help in
developing the Universal Feline Behavior Coding Scheme. We also
thank Michelle Sandau for her advice on creating the different solutions used in the study. Last but not least, thanks to all the staff at
AFB Internationals LRC ofce and PARC facility.
Appendix A. Supplementary data
Supplementary data associated with this article can be found,
in the online version, at http://dx.doi.org/10.1016/j.applanim.2016.
05.031.
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1

1

Supplementary material.
Feline Coded Behaviors for Drinking Tests

State Behavior = duration (indicate

Behavioral Events = occurs at a single

start and stop)

point in time

Sniffing and Drinking

The cat is not sniffing or licking at or drinking from the interior of
the cage or the spouts. No ingestion can be observed. Not
drinking or sniffing is also to be used when the cat is outside the
area of interest.

Not drinking or

Code: When the cat is more than 2.5 cm from something that it

sniffing

can sniff or drink.

The cat is ingesting liquid from the spout either via extending
its tongue and then retracting it in a lapping motion so that
the tongue comes in direct contact with the end of the spout
or by putting the end of the spout in its mouth and suckle
upon it.

Drinking from Spout

Code: At the first lick or when the mouth comes in contact

(ingestive)

with the spout.

Modifiers for Drinking from Spout

Drinking from spout

Ingesting from the spout located to the left from the

Left
Drinking from spout
Right

observers point of view.

Ingesting from the spout located to the right from the
observers point of view.
The cat is drinking from either the door or the
floor directly below the spout. The tongue is
extended and then retracted back into the cats
mouth. Whilst extended, the tongue comes in
contact with dripped solution on the surface of
the cage and results in ingestion.

Drinking dripped

Code: At the first lick or when the mouth comes

solution from floor or

in contact with solution on the door or floor just

door (ingestive)

below the spouts.

Modifiers for Drinking dripped solution from floor or door

ingesting
whilst the nose
is entirely in the
Drink dripped

left half of the

solution Left

cage.
ingesting
whilst the nose
is entirely in the

Drink dripped

right half of the

solution Right

cage.

The cat is sniffing the spout. The head and neck is visibly
extended and directed towards the spout or the dripped
solution. No ingestion is observed.
Code: When the cat is sniffing and within 2.5 cm of the
4

Sniffs Spout

spout.

Modifiers for Sniffs Spout

head, neck and nose indicate sniffing within 2.5 cm of the
Sniffs spout Left

left spout
head, neck and nose indicate sniffing within 2.5 cm of the

Sniffs spout Right

right spout
The cat is sniffing the interior of the
cage. The head and neck is
visibly extended and directed
towards a point in the cage. No
ingestion is observed.
Code: When the nose of the cat is
within 1 inch of the object of

Sniffs Other

interaction.

Modifiers for Sniffs Other

When the nose is
Sniffs Other Left

entirely in the left half

of the cage.
When the nose is
entirely in the right half
Sniffs Other Right

of the cage.

Eyes (during sniffing spout and ingestive behaviors only)

No Eyes (not

The cat is neither ingesting liquid nor interacting with the

recording)

spouts.
The eyes of the cat have a rounded shape and only slight
muscular contraction in the area around the eyes can be
observed. More than half of the full area of the eyes is visible
to the observer.

Eyes open 50-100%

Code: At onset of behavior.

The eyes of the cat are not fully open, rather the cat is
squinting. A contraction can be observed that narrows the eye
down to a slit rather than a rounded shape. The area of the eye
visible to the observer is then less than 50 % of the area that
can be observed when the cat has its eyes fully open.

Eyes open <50%

Code: At onset of behavior.

The eyes are closed and not visible to the observer forming
a solid line in the cats face as opposed a slit or a rounded
shape. The eye itself should in no way be observable. Only
when the eyes are completely shut the behavior for closed eyes
is to be coded.

9
10

Eyes closed

Code: At onset of behavior.

Eyes Undetermined

The eyes of the cat are not visible to the observer.

Ears (during sniffing spout and ingestive behaviors only)

11

No Ears (not

The cat is neither ingesting liquid nor interacting with the

recording)

spouts.
Both ears of the cat are perked upward, attentive, or forward.

12

Ears upward and/or

The ears form a 50 to 90 angle with the scalp.

forward

Code: At onset of behavior.

The bases of the ears are further apart than when forward;
possibly turned outward. The ears form an angle below 50

13

Ears lowered and/or

with the scalp.

outward

Code: At onset of behavior.

The ears are held in different positions compared to the scalp.
Example; one ear is in the upright position whilst the other one

14

Ears different

is folded outward.

positions

Code: At onset of behavior.

Behavioral Events
Flicks tongue quickly over the nose only. The tongue is
extended upwards and covers any part of the cats nose. The
tongue goes up to the nose and then back into the mouth
without continuing into a whisker lick. The mouth is generally
less open in a nose lick than in a whisker lick.

Licks nose

Code: At peak contact with the nose.

The cat sweeps its tongue to the side of its face (may start
as nose lick). The mouth is in most cases more open than it
would be in a nose lick. If the tongue is extended upward to

Licks whiskers

whiskers without the sweeping motion and without any nose

contact it is also to be coded as a whisker lick.

Code: At peak of behavior when the tongue is extended over
the whiskers or when the tongue comes into contact with the
whiskers without first sweeping over the nose.
The tongue is visible and directed forward out of the mouth
and not reaching upwards towards the lip or nose of the cat.
The tongue is not in contact with any object.
3

Tongue protrusion

Code: When tongue is visible.

The tongue is visible and directed forward out of the mouth

whilst the cat has its mouth wide open in a gape. The
tongue is not reaching upwards towards the lip or nose of the
Tongue Protrusion
4

Gape

cat. The tongue is not in contact with any object.

Code: At peak of behavior.
The cat performs a single chewing motion with its lower jaw
and might slightly open its mouth. Mouth smack is performed
without any form of ingestion. Mouth smack may, however,
follow directly after ingestion has been performed. During
mouth smack the cat may push its lip slightly outward.
Code: At peak of behavior. Each instance of mouth smack is to

Mouth Smack

be coded.

Miscellaneous

Miscellaneous Behavioral Events

2
3