Analysis of Nasutitermes corniger Termite Behavior in

Response to Varying Threat Levels

Yutaro Sakairi1, Katie Hunger2, Jiamei Shang2, Janis Shin3
1. Department of Environmental Science, University of Washington
2. Program on the Environment, University of Washington
3. Department of Bioengineering, University of Washington

Abstract
Eusocial animals demonstrate how individuals can perform specific tasks to
improve the colonys chances for survival. Although this may make the colony run more
efficiently, we have observed colonies overexert resources in response to a break in
their shelter, regardless of different stimuli presented. At the Cocha Cashu Biological
research station in Manu National park, we investigated whether Nasutitermes corniger,
colloquially known as conehead termites, were able to distinguish actual threats from
the coincidental presence of non-threatening insects near breaks in the termite tunnels.
We recorded the ratio of worker termites to soldier termites over time, for twenty
colonies in the presence of a predator, ant Ectatomma tuberculatum, and in the
presence of a non-predator ant, Atta columbica. Our results suggest that termites
respond in the same manner when presented with a threat or non-threat next to a
random break in the tunnel. This suggests that termites lack the complexity to respond
flexibly to distinct stimuli, which causes them to expend more time and energy in
defending their colony than necessary. Through our study we have concluded that the
pheromones of termite predators or lack thereof do not deter termites from rebuilding
their tunnels in the same locations.

Abstract (en espaol)

Los animales eusociales demuestran cmo las personas pueden realizar tareas
especficas para mejorar las posibilidades de supervivencia. Aunque esto puede hacer
que la colonia funcione ms eficientemente, hemos observada que las colonias fatiga
los recursos en respuesta a una interrupcin en su refugio. Esto ocurre
independientemente de los diferentes estmulos presentados. En la Estacin Biolgica
Cocha Cashu en el Parque Nacional Manu, investigamos si los Nasutitermes corniger,
Coloquialmente conocidos como termitas de "conehead" somos capaces de distinguir
las amenazas reales de la presencia fortuita de insectos no amenazantes cerca de las
roturas en los tneles de termitas. Registramos la proporcin de termitas obreras a
termitas de soldado a lo largo del tiempo, en presencia de una hormiga depredadora,
Ectatomma tuberculatum, y en presencia de una hormiga no depredadora, Atta
columbica. Nuestros resultados sugieren que las termitas responden de la misma
manera cuando se les presenta una amenaza o no amenaza junto a una ruptura
aleatoria en el tnel. Por lo tanto, sugiere que las termitas carecen de la complejidad
para responder de manera flexible a los estmulos, lo que les hace expandir ms
tiempo y energa en la defensa de su colonia de lo necesario. A travs de nuestro
estudio, hemos concluido que las feromonas de los depredadores de termitas o la falta
de ellos no impiden que las termitas reconstruyan sus tneles en los mismos lugares.

Introduction
Rather than having to perform all the functions of survival, such as defense, food
collection and reproduction, eusocial insects split the functions among their community.
This allows each individual to specialize in certain roles and allows the colony to
operate more efficiently. This high level of social organization encourages the formation
of altruistic societies that cooperate in order for the reproductive caste to thrive and
produce offspring (Tian and Zhou 2014). However, it is unclear what tactics eusocial
insects use to respond to threats, especially in cases when they must choose between
repairing their shelter or deterring a potential predator. Balancing resources toward
these two actions may determine the difference between survival and death for a
colony.
There is a limited body of research documenting the defense behavior of
eusocial insects in response to a predator. Eusocial insects are characterized by
multiple types of individuals within the colony, including workers, soldiers, and
reproductive castes. The roles of individuals also varied by age. A study on
Nasutitermes princeps found that soldiers patrol the source of the threat, older workers
approach the threat, whereas younger workers do not (Roisin et al. 1990). This
indicates that different castes and ages of individuals affect their roles when responding
to threats. Furthermore, while soldiers consistently responded aggressively to threats,
workers were found to behave flexibly depending on the composition of termites around
them, acting defensively when near soldiers and aggressively when near reproducers
(Ishikawa and Miura 2012). These results suggest that eusocial insects vary their
response depending on the makeup of the colony.
From studies conducted on other animals, we can deduce that many factors
impact antipredator behavior of animals, including genetics, past experiences, and
environmental conditions. A study on the antipredator behavior of 128 bird species
found that greater genetic variance correlated with a more extreme response to threat,
where the response was measured by the distance flown away from the threat (Jiang
and Moller 2017). This result indicates that antipredator behavior is at least partially
determined by genetics. Such responses to immediate threats may be hardwired in a
species DNA to instinctively react in a manner that protects them, allowing the species
population to keep existing. Some animals operate based on a landscape of fear, a
psychological tool used to determine antipredator behaviors (Yong 2016). A landscape
of fear is a learned geographical map that describes the perceived fear animals have
towards different locations based on their risk of predation. Rodents have been found to
change their foraging behavior based on past experiences, suggesting that antipredator
behavior can also be learned (Krijger et al. 2017). Additional studies focusing on four
herbivorous mammal species found that night time luminosity also impacts their
antipredator behavior. Mammals can form a landscape of fear based on light levels
and/or the lunar phase in order to decrease the chances of being spotted by a predator
(Palmer et al. 2017). The exhibited eusocial survival tactics of mammal species may
coincide within eusocial insects as well (Jennifer et al. 1994).
Our study expands on this body of research by analyzing the response to
different levels of threats by Nasutitermes corniger. N. corniger termites build nests on
the trunks of trees with thin tunnels connecting the nest to the ground. These tunnels
are made of a woody pulp that is easily broken. Upon breakage, worker termites swarm
to the opening to repair the tunnel. Soldier termites also arrive at the opening, guarding
or scouting the surrounding trunk. If a threat is found, soldier termites spray a sticky
liquid from their modified nozzle-shaped heads that hinder the mobility and irritate the
intruder (Eisner et al. 1976). For instance, a high number of worker termites and a low
number of soldiers might indicate that the disturbance is not seen as a threat.
Based on this physiological information, we wanted to determine whether the
termite composition and repairing efficiency of colonies were impaired by the presence
of predators at the tunnel opening. We hypothesized that the presence of a predator
would attract more soldier termites to fight it as well as deter the larger, more nutritious
worker termites. The absence of workers would also slow the reparation of the tunnel
opening. Based on this hypothesis, we predict that the presence of a predator will result
in a lower ratio of worker to total number of termites and decreased the amount of
opening repair.

Methods
To find the specific locations for our trials we hiked along trails 4 & 6 which
stretched between a quarter and a half mile from Cocha Cashu Biological Station,
located in Manu National Park, Madre de Dios, Peru. We first marked nests located
near the trail system that were active. Activity was determined by cutting a hole in the
vein of a tunnel. We assumed it was active if more than five termites exited the tunnel.
We chose twenty nests altogether and performed three trials over a four-day period on
each nest. We tested each nest around the same time each day to reduce bias.
We chose a natural predator of N. corniger found in the Amazonian rainforest
near Cocha Cashu Biological Field Station, Ectatomma tuberculatum. We also used
leafcutter ants (Atta columbica) as a control group, as it does not prey on N. corniger.
Comparing results from a predatory and non-predatory species of ant would ensure that
our results were generated not purely from the fact that we were introducing an ant into
the vicinity of the opening, as opposed to a threat to the termites. To secure the ants
near the opening, we considered crushing the ants and smearing their contents onto the
trunks of trees because termites primarily communicate and interact with the world
through chemical smells (Costa-Leonardo and Haifig 2013). However, this smearing
method would impair our study as we would be testing the same nests multiple times
and smears from previous tests would impact our results. In addition, we were not
entirely sure about how termites detect ants, whether that be through smell, vibrations,
or some other senses. Therefore, we decided it would be most reliable to secure the
whole living ant to the tree trunk with a pin.
The night before each test day we went out and collected predator ants and leaf
cutter ants to ensure that they were alive until the moment we used them in our
experiments, so that their scent would not fade away. As a control, we only selected
larger, soldier ants as they were naturally more likely to predate termites. For the
predator ant trial and leaf cutter ant trial, we pierced through the ant with a pin and stuck
it on the bark next to the tunnel. Then we cut a 1.5cm hole in the tunnel, took a picture
of the initial reaction of the termites, and continued to take a picture every minute for 20
minutes. These pictures allowed us to count how many workers and soldiers were
visible. For our pure control trial, we did not place anything next to the vein before
cutting a 1.5cm hole. We also randomized the trials for each nest so we did not do the
same trial on all the nests each day. This was to prevent order bias from tainting our
results, eliminating the possibility that colonies might be reacting differently based on
the number of days we repeated the experiments rather than which trial we were
testing.

Results
Fig. 1 demonstrates the ratio of workers to total number of termites across 20
minutes. None of the treatments finished repairing the hole within 20 minutes and all of
the treatments showed very similar results. We ran a T-test for each minute from time 1
to time 20, comparing control vs non-predator, control vs predator and non-predator vs
predator. The lowest p-value generated was 0.08, which demonstrates that the data are
not significant at any time.
 Fig. 2 shows the percentage of colonies that exhibited different defensive
behavior in the first twenty minutes after the opening. The guarding behaviour indicates
that termites in the colony were lining up around the outside of the hole cut into their
tunnel. This behavior is a defensive strategy to ward off predators while the workers
travel back and forth to repair the tunnel. The attacking behavior was characterized by
one or more termites approaching the predator or non-predator ant to spray them with
glue in order to scare them off or secure them in place. The scattering behavior was
displayed when more than five termites wandered more than 1.5cm outside the tunnel
boundaries, presumably to search the area for threats. None of the distribution of
behaviors observed across colonies was significant with the smallest p-value being
0.434.
Discussion
Based on our results, termites did not appear to distinguish between the varying
threats introduced to the opening of the tunnel. We came to this conclusion from Fig. 1,
where the trend line of each trial increased linearly at a similar rate. This demonstrates
that workers tend not to approach potential threats and repair the tunnel only after
enough time has passed for the threat to have been eliminated. On the other hand,
soldiers defended the colony by swarming near a break in the tunnel after a break was
detected and retreated as repairs to the tunnel began.
After analyzing the observed termite behavior across all three trials in Fig. 2, we
concluded that there was no significant difference between the behavior of termites
when a predator ant, non-predator ant, or the control was present next to the tunnel we
opened. This behavioral analysis expands on our conclusions from Fig. 1 by showing
that neither the composition nor the defensive behaviors of N. corniger termites vary in
response to a potential threat.
Several causes can be attributed to how termites do not alter the composition of
castes and their behavior in response to varying levels of threats. One potential cause
could be that N. corniger termite colonies have not needed to evolve differentiated
responses to holes in their tunnels. For instance, there may be no predator of N.
corniger in the rainforest around Cocha Cashu Biological Field Station that have
threatened their survival by exploiting their defensive behavior. Another probably cause
is that the brains of N. corniger are not complex enough to process differences in threat
levels when their tunnel is broken, communicate this with the rest of the colony, and
alter their group behavior. In contrast, our literature review found that some species of
birds and mammals alter their defensive behaviors based on environmental factors and
past experiences. This makes it clear that other animals are capable of responding
more dynamically to threats, where brain size may be the determining factor.
Currently, the issue of N. corniger extermination has grown as they have
migrated to Florida and caused extensive property damage (Scheffrahn 2013).
Identifying specific mechanical behavioral triggers in termite colonies will enable us to
find easier and safer ways to eradicate termites. For instance, exterminators may be
able to take advantage of attacking behaviours to draw out more soldier termites by
placing dummy ants outside of their tunnels if flailing insect bodies are indeed triggers
for termites to attack. Such methods could save money and possibly limit the use of
toxic extermination chemicals.
In future studies, we would like to know whether the number of ants (Garvey
2009) put next to the tunnels makes a difference in the termite response since it would
alter the concentration of pheromones from ants. Furthermore, because the smells that
exude from living or dying creatures are different (Walker 2009), next time we would like
to control for the liveliness of the ants.

Acknowledgements
University of Washington, Program on the Environment
Cocha Cashu Biological Station staff
Ursula Valdez
Tim Billo
Marc Seid
Eric Delgado

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