Aggression Based on Feeding Schedule

Zachary Lenzi
Introduction:
In my bedroom, I have a 10-gallon planted fish tank that houses a Crown Tail Male Betta
Fish and an African Dwarf Frog. They have been kept together in this tank for over two months,
and during this time I have noticed that when I fast them for a day, they tend to nip and bite at
each other during the next feeding day. This led me to the question, “does a decreased feeding
schedule lead to an increase in aggression between these two species?” The African Dwarf Frog
has been determined to have a peaceful behavior while the Betta Fish has a semi-aggressive to
aggressive temperament, which is usually only towards females and other fish with the same
body type as it. From this observation, I posed the question, "Does altering their schedule of food
cause aggressive interactions between these two specific organisms?” This led me to hypothesize
that it would cause a change in aggression. Based on this hypothesis, I predict that when I
decrease the feeding schedule from daily to alternate days, there will be an increase in aggressive
interactions between these two species. I will test this hypothesis over three weeks to determine
if there is a relationship between the schedule and behavior.
Methods:
The setup that was used for this experiment was in my bedroom out of direct sunlight
from the window and the study lasted over a period of 18 days. The equipment that is used in this
experiment was a 10-gallon Aqueon fish tank that has been cycled and properly stocked with live
plants and gravel substrate. The lighting that was used was an Aqueon Hang Over Plant Light
attached to an outlet timer that has it run from 8 am-4 pm every day. The heater and filtration are
both Tetra brands and are the appropriate size for the tank. Both the African Dwarf Frog and
Betta Fish were fed the same food, Omega One Freeze-Dried Blood Worms, with four being fed
to each subject. A bubbler with a small air stone also ran in the corner of the tank away from the
filter to create better circulation. Having the light set to a timer and the heater and filtration
keeping the water parameters constant during this experiment, the only input that was being
affected was the food supply. This allowed me to single out one factor directly instead of
worrying about outside, confounding factors skewing the data.
This manipulative experiment was conducted over a period of 18 days. These days were
separated in increments of three days of daily feeding followed by three days of feeding on
alternate days. Only 12 of the days are going to have recorded data. This is because the first day
of each test type is going to act as a buffer. On feeding days that are every day, they are going to
be fed normally. On alternate day feedings, the first day is going to have no food. This will act as
a buffer day so that the transition between every day to alternate days will always have one
period of no food. This process will be repeated six times until there is a total of three daily
feedings followed by three alternate day feedings, causing a total of three replications. Feeding
will occur at 8:30 am every day with observations being examined until 9 am. This totals 30
minutes of observations. The response variable that is being recorded during this period is
aggressive interactions between the two individuals. This is either if the betta fish flares his gills
or nips at the frog or if the frog attempts to bite the betta fish. This data that is being recorded is
continuous since its total daily attacks. Since the experiment is conducted in the following way,
it will be a trajectory experiment with the responses tracked over time.
With the variables stated above that are being tested, the correct statistical test to analyze
the data would be a paired T-Test. There is only one factor that is being tested during this
experiment, which is the number of aggressive actions in relation to the feeding schedule. Since
the test groups are not independent of each other, the one-tailed paired T-Test would be the best
option if the data is significantly different from each other. The alpha level that I used for this
experiment is 0.05 and the statistical analysis will be performed in RStudio.
Results:
After the completion of the data collection, I created a line graph from the data to
compare how the data relate to each other on a six-day continuous scale (Figure 1).

Figure 1: Line graph comparing the data of the two treatment types. This graph is altered to represent if each treatment lasted
over the course of six continuous days to examine variation.

The first statistical test that was performed on the data was the Wilcoxon signed ranked test with
continuity correction (Table 1). This was used to determine if the two categorical independent
variables (everyday feeding and alternate day feeding) are significantly different from each
other. This was assessed based on an alpha level of 0.05. The two groups were determined too
not be significantly different from each other. Even with that concluding the results, a one tailed
T-Test was performed afterward which was also came back not significant, which was expected
(Table 2).

Wilcoxon Signed Rank Test with Continuity Correction

Data: Every day and Alternate Days
V=6 P-Value = 0.7835
Alternate Hypothesis: True Location Shift is not Equal to 0
Table 1: Wilcoxon Test of Every day and Alternate Day data comparison, based on an alpha level of 0.05.
 Paired T-Test
Data: Every Day and Alternate Days
t = -0.45964 Df = 5 P-Value = 0.6675
Alternate Hypothesis: True difference in means is greater than 0
95 percent confidence interval:
-2.691992 Inf
Sample estimates:
Mean of the differences -0.5
Table 2: One-way Paired T-Test with Every day being the baseline and checking if the alternate day treatment is checked for
lateral movement.

Discussion:
From the data that was obtained from the Wilcoxon Signed Ranked Test and the One-
Tailed Paired T-Test, it can be concluded that I failed to reflect the null hypothesis. This is
because the P-value that was obtained from the Wilcoxon test was 0.7835. This number is
significantly greater than the alpha level of 0.05. This means that the data is not significantly
different and that no further tests need to be conducted. Even though this was the case, one tailed
t-test was still performed to confirm the results which were found. This can conclude that the
aggressive interactions between the Betta Fish and the African Dwarf Frog are not caused by a
decrease in feeding schedule. The experiment did not have any confounding factors found during
the experiment. The temperature, water quality, and lighting all remained consistent throughout
the entire experiment. The only variable that was changed was the food source schedules.
The main factor which could have affected the experiment was its design. When figuring
out how to reduce carryover from one treatment to another, one day was assigned at the
beginning of each treatment to act as a buffer. This was set to one day because of the time
constraint which was on the experiment. Theoretically, it would have been preferred to have a
week of buffer and then a week of recorded data for more precise results. Another factor that
could have affected the experiment is that only one aquarium was being treated. This makes it so
that the data collected is never truly independent of each other. Another aquarium with the same
equipment and plants would have been preferred, but then other factors would have complicated
it more. This would be the individual Betta Fishes personalities which vary between fish, even if
they are the same body type and color. Further tests with different varieties of Betta Fish would
be interesting to examine to determine if variety would influence their aggression.
Appendix: