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Associative learning capability of the

cricket Acheta domesticus in the


context of food choice
Nicole Duquette, Laura Fox, and Michael
Zwick
Department of Psychology, Neuroscience, and Behaviour;
McMaster University
Several studies have documented the excellent associative
learning abilities of the field cricket (Gryllus bimaculatus) (Matsumoto
and Mizunami 2000, 2002a, b, 2004, 2006), but comparatively much
less is known about learning in the house cricket (Acheta domesticus).
We investigated the house crickets olfactory, visual, and spatial
associative learning capability. Crickets were trained to associate one
of a pair of odours, colours, and spatial locations with appetitive cricket
food, and the opposite cue from each pair with cricket food that was
laced with an aversive substance. 3 days after, crickets were tested
using two non-aversive food dishes with the same sets of cues from
training. Crickets did not exhibit a preference towards the dish with
the cues that corresponded to the appetitive dish in training.
Therefore, we cannot conclude that house crickets are capable of
associative learning.
INTRODUCTION:
Though it was once thought that insects could respond to
environmental stimuli only with fixed, innate behavioural patterns
(Wheeler 1930), research over the past few decades has proven
otherwise. Associative learning, defined as the ability to form an
association between a state/event with a certain stimulus (Dudai 1989), is
exhibited by a wide array of insect species. Such ability is especially
advantageous in the context of food choice. Insects must often decide
which food source to exploit from a number of different options. Being
able to discriminate the quality of food sources not only helps an insect
improve its nutrition and avoid illness, but it also makes foraging more
efficient.

The field cricket is an example of an insect whose associative


learning abilities are firmly established in the literature. They are able
to associate one odour with water reward, and another odour with an
aversive saline solution (Matsumoto and Mizunami 2000; 2002a, b;
2004; 2006). Field crickets can also use visual cues in conjunction with
olfactory cues in order to learn. It has been documented that they can
associate an odour with water in the light and saline solution in the
dark, and a second odour with water in the dark, and saline solution in
the light (Matsumoto and Mizunami 2004). Crickets are also able to
learn to locate a cool spot in a heated arena by using visual cues from
the environment (Wessnitzer et al. 2008). Taking these findings
together, it appears that field crickets use associative learning to seek
out appetitive stimuli and avoid negative stimuli, and they do this
extremely well when they can make use of olfactory and visual cues.
What remains largely unknown is whether the house cricket, a
related species, shares this excellent associative learning ability.
Scotto-Lomassese et al. (2003) found that the house cricket was able
to use olfactory and/or visual cues to choose a hole that led to escape
over one that was a trap. However, to our knowledge, there are no
other learning studies using the house cricket. To help rectify this, we
conducted our own investigation on the topic.
We wanted to see if house crickets could learn to associate one of
a pair of odours, colours, and spatial locations with an aversive food
dish, and the opposite set of cues with an appetitive food dish. We
know that the field cricket has colour vision (Zufall et al. 1989), so we
are making a crucial assumption that the house cricket does too. We
predicted that during testing, the crickets would significantly prefer an
appetitive food dish whose cues corresponded to the appetitive food
dish from training over an appetitive food dish that was associated
with aversive cues.
MATERIALS AND METHODS:
Preliminary Preparation
Approximately 200 house crickets were obtained from a
commercial supplier and housed in a large aquarium where food and
water were freely available. Next, we wanted to confirm that the two
spices we planned on using in our experiment (cinnamon and cumin)
were not offensive to the house cricket. We selected 10 crickets at
random, transferred them into individual containers, and placed two
dishes of cricket food at opposite sides. One dish was mixed with
cinnamon and the other with cumin, and we counterbalanced the sides
of each kind of dish. We found that crickets readily ate from both

dishes, and thus determined that cinnamon and cumin would be


acceptable to use in our experiment.

Figure 1. The combinations of CSs used in each of the 4 containers.

Training Phase
60 adult female crickets were selected at random from our
aquarium and were equally distributed into four large containers. Each
container represented a distinct condition. A summary of the
conditions is displayed in Figure 1, and the experimental setup is
depicted in Figure 2a. In each container, the sole food sources were
two food dishes, across from each other. Both dishes were composed
of commercial cricket food. One dish was 1% quinine, a substance
aversive to the cricket (Matsumoto and Mizunami 2000), while the
other did not contain quinine; these served as the aversive and
appetitive unconditioned stimuli (US), respectively. Each dish also
contained either cinnamon or cumin, in order to give each food dish a
distinct odour. 10 grams of spice was used for every 100 grams of
food. One of a pair of odours (cinnamon/cumin), food dish colours
(brown/green), and spatial locations (left side/right side) were used as
conditioned stimuli (CS) for each of the two dishes.
The only combination of CSs that remained constant throughout
every condition was that brown food dishes always smelled like
cinnamon, and green food dishes were always associated with cumin.
We counterbalanced in 3 ways. Half of the crickets were trained with
the appetitive US on the left side and the aversive US on the right side,
and vice-versa. Cinnamon served as a CS for the appetitive US for half
of the crickets, but was a CS for the aversive US for the other half, and
the same was true for cumin. Similarly, the cinnamon-scented food
was on the left side for half the crickets, and on the right side for the
other half; again, the same was true for the cumin-scented food.
For 7 days, the crickets were allowed water and fed this diet ad
libitum. Then we randomly selected 8 crickets from each container
and moved them into individual containers. The crickets were starved
for three days in order to enhance their motivation to search for food in
the testing phase.

Testing Phase
Each trial began when we placed two food dishes into each
container across from each other at one end. Other than the fact that
neither food contained quinine, the cue combinations were consistent
with those from training. Figure 2b illustrates the experimental setup.
Crickets were allowed 15 minutes to eat from one of the two dishes.
Two blind observers recorded which of the two dishes each cricket
initially chose to eat, and we considered this dish to be the one that
they preferred. If the cricket did not eat within 15 minutes, we
considered the cricket to have made no choice, and did not include
these data points in our statistical analyses.

Figure 2. A simplified version of the experimental setup. Note that the specific combination
of stimuli presented is unique to one of the 4 conditions. a. Crickets were trained to
associate one of a pair of odours, colours, and spatial locations with food containing quinine,
and the other pair member with non-aversive food. Note that the cricket in b. was trained to
associate cinnamon, brown, and the left side with quinine this was done to better illustrate
the relationship of the stimuli presented in both phases of the experiment. There were also
15 crickets and pieces of egg carton in each container, but they were omitted for clarity. b.
The stimuli presented to each cricket differed from that from training solely in that neither
dish contained quinine. Egg cartons were also present in the cage, but were omitted for
clarity.

Data Analysis
A binomial test was used to determine if the proportion of crickets
that preferred the dish that had appetitive cues was significant. We
used a Fishers Exact Test to determine whether there was a significant
difference in performance depending on the combination of CSs with
which a cricket was trained. Two more binomial tests were conducted

to determine if the crickets had a significant preference for a particular


spice or side.
RESULTS:

Figure 3: Crickets were trained to associate an odour/side pairing with an aversive dish, and
another pairing with an appetitive dish. Possible combinations of CSs associated with the
aversive US were cinnamon and the left side (Cinnamon/L -), cinnamon and the right side
(Cinnamon/R -), cumin and the left side (Cumin/L -), and cumin and the right side (Cumin/R
-). Shown are the proportions of crickets in each condition (and overall) who preferred the
dish that had the appetitive cues (proportion learned).

4 of the 32 crickets died before testing and 2 did not make a


choice. 17 out of the remaining 26 crickets preferred the dish with
appetitive CSs, but this preference was not significant (z=1.37,
p=0.1686; Fig. 3). There is no evidence that performance differed
depending on the combination of CSs with which the crickets were
trained (P=0.3863; Fig. 3). The crickets did not have a significant spice
preference (z=0.98, p=0.3269), nor did they significantly prefer one
side over the other (z=0.59, p=0.5572).
DISCUSSION:
We did not find evidence that house crickets are able to associate
odours, colours, or spatial locations with appetitive or aversive food
dishes, and this was not due to an innate spice or side bias, or due to
the combination of CSs the crickets were trained to find appetitive.
This is incongruent with the Scotto-Lomassese et al.s (2003) work with
house crickets and Matsumoto and Mizunamis (2000, 2002a, b, 2004)
series of experiments with field crickets. There are several possible
explanations for this.
For one, we lost 6 crickets between training and testing. As a
result, our sample size was below what was necessary in order to
achieve sufficient statistical power. So it may the case that the null
hypothesis was indeed false, but we simply cannot prove it.

It is also possible that there were problems with the preparation


of our food dishes. Studies similar to ours used saline solution as the
aversive US and water as the appetitive US with diluted artificial
essences as odorants (Matsumoto and Mizunami 2000; 2002a, b;
2004), whereas we used spiced cricket food with and without quinine,
respectively. However, quinine may not have been a good choice:
Matsumoto and Mizunami (2000) chose against using it in their
experiment because of its tendency to cause illness. The crickets may
have become ill from some quinine consumption during training, which
interfered with their learning. This idea gains support from the fact
that 3 crickets died during the starvation period. It is also possible that
the concentration of spice in the food was too low, and thus the odours
were not salient enough for the crickets, leaving them unable to use
the olfactory cues. Therefore, a follow-up study should be conducted
using water and saline solution as the USs and artificial essences as
the CSs.
It is also worth noting that it remains unclear whether crickets
can use spatial cues in the way that we thought. We hypothesized that
they could learn to associate one side of the container with the
appetitive US and the other with the aversive US. However, it appears
crickets use the visual cues associated with a spatial location, as
opposed to the location itself (Scotto-Lomassese et al. 2003;
Wessnitzer et al. 2008). In our study, the containers that housed the
crickets were in different areas in the lab during training, starvation,
and testing. As a result, the visual cues associated with the left and
right sides of the container changed 3 times, and thus, it was
impossible for the crickets to use this type of cue.
We must also take into account that the experimental procedure
used by Matsumoto and Mizunami (2000; 2002a, b; 2004) and ScottoLomassese et al. (2003) differed from ours in an important way. In all
of these studies, both the appetitive and aversive US were present
during testing, but in our experiment, the aversive US was not present
in the testing phase. This suggests that our results may be consistent
with Hoffmanns (2003) anticipatory behavioural control framework
which maintains that behaviours are performed in order to attain a
desired outcome. During training, one odour, colour, and spatial
location signalled the presence of quinine, whereas another trio of cues
suggested its absence. If quinine was present during testing, crickets
would have benefited from using cues to track down the dish without
quinine. However, quinine was not present, so it is reasonable that
they did not spend time trying to avoid something that was not
actually there. Generally speaking, if the aversive US is absent,

aversive memories need not be behaviourally expressed (Gerber and


Hendel 2006), because the desired outcome (in this study, avoiding
quinine) would be attained regardless. For this reason, in follow-up
studies, we should have the aversive US present during testing.
Overall, the associative learning abilities of the house cricket are
still not fully understood. Though it is plausible that they are indeed
good learners, just like field crickets, our results are not consistent with
this point of view. Considering that there is only one other study that
examines learning in house crickets (Scotto-Lomassese 2003) and it
had opposite results to ours, it is clear that additional studies are
necessary to resolve this inconsistency.
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