Light Induced Sleep Deprivation in the common Rosy Red Minnow

(Pimephales promelas)

Kari Mueller & Eric Schmit

Loras College, Department of Biology
1450 Alta Vista St
Dubuque, Iowa 52002

To determine the effects of sleeplessness in fish on reaction time to feeding,

appetite, and reaction time to predators

ABSTRACT
A proper light cycle is believed to be crucial to a captive fishs health and longevity. Using
the common rosy red minnow (Pimephales promelas) from the fat-head minnow family, we
exposed two schools to controlled and variable lighting conditions, testing for feeding
reaction times, aggression, and the amount of food eaten between the two schools. We also
observed behaviors such as schooling and hiding. A proper 12 hour on, 12 hour off cycle for
our control group did show to be beneficial to reaction time for feeding and less aggression
throughout the experiment; compared to our test group like we had predicted. For the test
group, we set up a light cycle of 15 minutes on and 15 minutes off repeating and later
switched to a blinking mechanism. However, testing for differences such as the difference in
the amount of food eaten between the two groups suggested there were no differences
according to an independent samples T-test. We observed but did not test that the control
group showed tighter schooling and more hiding behaviors than the test group.
INTRODUCTION
A proper light and dark cycle does many things for an aquarium. Enough darkness keeps
green algae from over populating the aquarium, while light allows for aquarium plants to
grow, and more importantly, allows fish to see, encouraging them to be active. However,
many aquariums do not have a normal light and dark cycle, which could potentially stress
the inhabitants inside, such as being used to slap a Swede (NYKOPING, 2013). A poor light
cycle can be attributed to by lights in the room of the aquarium frequently going on and off
as people enter and leave the room, when aquarium lights are turned on for short viewing of
the aquarium, or from increased and decreased lighting from the sun. These may be
especially stressful when fish are sleeping, as fish have no eyelids and are sensitive to lights
suddenly coming on and strong light intensity.
In researching other studies on the effects of sleep deprivation in fish, we found that sleep
deprivation may be followed by a large increase in activity, followed by low activity and

reduced rest. (Tobler & Borbely , 1985). Another study concluded that fish have ridged
diurnal feeding patterns (Chen, 2007), this means that the majority of feed prefer to feed
while light is present and constant. Using this information, we predicted that a fishs
reaction time will increase from and continue to over time, and that appetites will decrease
due to the stressful impacted-sleep environment. We also predicted that casualty rate in the
sleep sleep-deprived environment will be higher than the control due to the high-stress.
Fatality rate was predicted based on prolonged sleep deprivation in mammals causes death
(Rechtschaffen, 1983)
Throughout our experiment we tried to discover what the disruption of sleep patterns by
light does to aquarium fish, using Rosy Red Minnows. We tested for a difference in reaction
times from being fed, the amount of food eaten, the number of acts of aggression, and
schooling size between test and control tanks exposed to proper and improper lighting. We
observed behaviors such the spread of the school and location of the school for each tank,
for which we expected the test group to congregate at bottom or darker areas of the tank
(Singh, 2013). We used statistical analyses such as an independent samples t-test, linear
regression, and correlation tests to measure the difference and see if there was a significant
difference in each group. We formed a hypothesis that the sleep deprived group of fish
would have a significant difference in reaction time to food, more counts of aggression,
consume less food, and attempt to hide more often when compared to the control group.
METHODS
The experiment performed took place in three ten-gallon glass aquariums provided for use
by the biology department of Loras College in Dubuque, Iowa. The tanks were lined up side
by side, with the two outside tanks becoming the control and test tanks, and the middle tank
becoming a common sump. Both the outside tanks had a one-inch inside diameter vinyl
tubing acting as a syphon to the middle sump, and a 77 gallon-per-hour pond pump
supplying water to the test and control tanks from the sump via two three-eighths inch
sections of vinyl tubing. Each of the outside tanks contained an air stone with air supplied by

a common air pump, and two broken flat pieces of terra-cotta to house beneficial bacteria.
The sump was provided with many pieces of terra-cotta to house more bacteria from an
established tank. Nitrifying bacteria broke down toxic ammonia from the fishs waste and
turned it into less harmful nitrates and nitrites. Two nylon mesh sacks filled with carbon were
also placed in the sump to absorb excess water chemicals such as heavy metals and
chlorine from tap water. Each of the outside tanks were darken out by covering the glass
with cardboard, which could be flipped up for viewing, and a standard aquarium lid equipped
with two 15 watt lights was placed on top. A diagram of this system is shown below in figure
1.
Figure 1: System set up

The system was allowed to stabilize before adding any fish. Once the system had been
running for several days, 18 Rosy Red Minnows were added to both the control and test
tanks, keeping both tanks under normal lighting (12 hours on, 12 hours off) while the
populations stabilized. This was done so that the death of a fish due to a change in
environment would not influence our data and be recorded as a death due to stress of sleep
deprivation. Once stable populations were achieved, the control tank was set on a timer to
continue the 12 hour light/12 hour dark cycle, and the test tank was set up for a hour
light, hour dark cycle using outlet timers.
From this point on data was recorded. When one week was left of the experiment, the timer
for the test group was exchanged for a flasher which would turn the lights on and off every

two seconds. Data observations included feeding time reaction, the amount of food offered
compared to amount of food eaten, deaths, signs of aggression, and school size for. We were
able to measure food offered and consumed within a 5 minute period by using a processed
flake food by Tetra brand with approximately equally sized food disks and a timer.
Aggressions signs counted as any clearly observed chasing, fin, tail, gill, or mouth nipping
within a three minute period. A log was also kept of information we felt useful such as where
the minnows may be schooling, deaths, other behavioral information, and any events that
may influence the results of this experiment.
RESULTS
When data started being collected after system stabilization with fish there were 16 fish in
test school and 15 fish in control school. To analyze the data we placed all
measured/observed data into SPSS to calculate results and determine if our recordings for
reaction time, death, feeding, and agression were statistically significant and therefore,
supporting our hypotheses. We performed a independent samples t-test on the reaction time
to feeding. We chose to use a independent samples t-test to verify the overall effect of
sleep deprevation due to the nature of recording and hypothesis paired samples t-test would
be innapropriate.
The results of the t-test gave us p=.021. Since our p<.05 was significantly different this tells
us there was a significant difference between the two groups shown in Figure 1. Our next
step was to perform a t-test on ammount of food eaten to prove Chens 2007 study that
fishs appetite decline due to sleep deprivation. This data showed no significant difference
p=1 explained further in discussion . The next observation of deaths within each group
proved to have no significant data with 1 fish dead in each sample. The last observation of
Agression we used a independent samples t-test and calculated a p=.015 again giving us a
significant difference between the two groups. During the experiment we observed a
difference in the behaviour of schooling between the two groups. So in response to the

observed behavior the Last third of the experiment we began to arbitrarily measure the
schooling size of each fish tank in relation to behavior.

DISCUSSION
This experiment found significant result (P=.021) supporting that mean reaction time of
being fed for control was approximately 3 seconds which was significantly faster compared
to the test at approximately 7 seconds. The results are also consistent with our hypothesis,
however not comparable to the previous study of Tobler and Borbely, who found that sleep
deprivation may be followed by a large increase in activity and then by low activity. Our
studies suggest that our fish had a decrease in activity at first, but later adjusted to near
normal activity.
In our predictions we said the diurnal feeding pattern and the interrupted sleep would cause
fishs appetite to decrease. By monitoring food intake we expected to see the fish neglect to
eat some of what was offered but we gathered no significant data. Every day the fish ate all

the food offered to them within 5 minutes. These results could have occurred because we
fed them once a day with some variance in time of day.
We also predicted that sleep deprivation would cause more overall stress, leading to more
deaths in the test group. This was based on the human physiological response that extended
sleep deprivation results in death. Since humans die from extended sleep deprivation we
expected some fish to mirror this response. Our observations totaled one fatality in each
group; in conclusion these results suggest that most fish were able to adapt to the disrupted
light cycles and created tolerance. The stress was not fatal. Test fish became tolerant of the
light disruptions after approximately 15 days according to observations and thus no
significant difference in death was observed.
The last prediction observed and tested is aggression. We expected that a fish under stress
would show more signs of aggression then a fish with lower stress. Fish were observed for a
3 minute person for any fin/gill/or tail nipping and chasing. Statistically our data showed a
significant difference (p=.015). The mean values for the control tank averaged at .3
compared to the test tank at 1.2 observed aggression signs within the time period. This
means that the control tank was statistically less aggressive then the test tank supporting
our hypothesis that sleep deprivation may cause aggressive behavior in fish. There was only
15 data points recorded in the aggression testing as we did not search for aggression every
day. Aggression was haphazardly measured by two observers which could cause inaccuracy
and imprecision in the data watching for fin nipping and chasing behavior. This particular
variable may be hard to repeat in following studies but should be repeated in order to
validate the results.
Aggression specifically should be refined further by defining aggression signs more explicitly
such as attacking the gills, mouth, or fins perhaps even quantifying the type of aggression.
Further improvement could be made by lengthening the observation period or observing
more frequently such as 2-3 times daily instead of one. By doing this more data could be
collected therefore lessening the possibly of the data occurring by chance.

In conclusion intermittent light patterns causes increased activity and then low-normal
activity with no change in appetite or fatality rate. It does however appear to cause
increased signs of aggression as supported by our p<.05 and change in observed behavior
such as schooling.

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