Olivia Liang - Animal Behavior Lab Report

Drosophila melanogasters Negative Behavioral Response to Multiple Treatments

Abstract
The purpose of this Animal Behavior lab was to analyze the behavior of Drosophila
melanogaster, which permitted researchers to gain a better understanding of the complexity of
human biology through the study of this model organism. This study explored the question, how
will different application of treatments, such as light, heat, apple cider vinegar, caraway and the
angle of the apparatus affect the number of D. melanogaster in each quadrant (A, B, C and D) of
the apparatus? 2, 6 and 10 minutes after D. melanogaster were transferred into the apparatus from
the original vial, the number of D. melanogaster were counted and recorded in each quadrant.
After individual results were collected, the class collectively pooled data. Finally, the chi-squared
statistical test was performed with each treatment test. Consequently, based on the chi-squared
statistical test results, the null hypothesis for all treatment tests were rejected because the results
were all significant with a p value of less than 0.05.
1. Introduction/Background
Drosophila melanogaster, a specie of fruit fly, is a segmented arthropod that consists of a head,
thorax and abdomen. D. melanogaster are produced in large quantities that permits them to be examined
with ease. It has a lifespan of approximately two weeks, and its life begins as a form of a fertilized egg.
Then, the fertilized egg hatched into a larva that changes physically through larval instar stages
(gastrulation, hatching, pupation) until it undergoes metamorphosis (called holometabolous) and develop
into an adult form. During their short lifetime, females eggs are laid in necrotic material and these eggs
are developed externally (Markow 2015). From a fertilized egg, D. melanogaster develops into a
multinucleated embryo during cleavage and a gastrula during gastrulation within 2-3 hours after
fertilization. Then, 16 hours after fertilization, D. melanogaster is developed into a segmented embryo,
and 8 hours later, it becomes a feeding larva through hatching. 2 days later, it becomes a pupa through
pupation, and it takes 4 additional days to become gametogenesis through metamorphosis. From there,
female and male mate and female reproduces. The fertilized egg then undergoes the same lifecycle as its
parents (Wood 2015). Due to the short life span of D. melanogaster, within one week, it reaches its stage
of sexual maturity and female reproduces sexually. Also, the males 7-tricosene, a pheromone, restricts
males sexual behavior by reducing homosexual relationships within the population; however, it does not
play a role in females mating behavior (Grillet et. al 2015).
D. melanogasters importance involves its purpose as a model organism due to their short
generation time, its easiness to manipulate and its large brood size. The importance of studying the

Olivia Liang - Animal Behavior Lab Report

behavior of this model organism allows researchers to deepen their understanding of the branch of human
biology and biological diversity (Wood 2015).
It is important to quantify the behavior of D. melanogaster to determine the specific factors that
influence the extent to which it responds to external factors and how its behaviors are shaped by its
genetic and environmental factors. The adult mating behavior between female and male D. melanogaster
begins with the exchange of visual cues and sensory stimuli that led to sexual attraction. Being able to
understand the courtship behavior of D. melanogaster permits the discovery of their genetic basis and
activities that shape their life and structure their influence on the environment (Nichols 2012). Also, large
quantities of D. melanogaster species are often found on rotting matter, such as fruits and vegetables
because the microorganisms on the rotting matter are part of their diet and contribute to their health
(Chhabra 2013).
In addition, this study proposed the question, how will different treatments, such as light, heat,
apple cider vinegar, caraway and the angle of the apparatus affect the number of D. melanogaster in each
section (A, B, C and D) of the tube? Then, the research question was tested with an alternative hypothesis
that states the differences in treatments will have an impact on the behavior of D. melanogaster and the
location of their position inside of the apparatus. Based on the hypothesis, a prediction was made: the
treatments, such as light, heat, apple cider vinegar, caraway and angle of the apparatus will cause D.
melanogaster to move away from the treatment and toward the direction of quadrant A.
Because this study investigates the behavior of a model organism, D. melanogaster, the discovery
of how the biology of a single organism works allows researchers to apply their understanding of a single
and simple organism to a more complex organism, such as human. This action defined the importance of
this study.
2. Methods
Prior to the animal behavior lab, the D. melanogaster were propagated to ensure that there were
enough D. melanogaster to work with in 4 weeks. In order to propagate the D. melanogaster, D.
melanogaster were transferred from an old vial via a funnel into a new vial that contained the same
media, such as 5 granules of baker's yeast and one netting. Then, both vials were plugged with a foam and
were sealed with a cap. Lastly, the vials were labeled with the group members names, current date and
indicated if the vial is new or old with using a sharpie. In two weeks, these step were repeated by
transferring D. melanogaster from old and new vials into two new vials separately, and each new vial also
contained the same amount of media as before.
Two weeks later, the experiment on the behavior of D. melanogaster begun. The apparatus was
built with using two clear tubs, two end caps, one T-joiner and one reducer, and it was divided into four

Olivia Liang - Animal Behavior Lab Report

sections and each section was labeled as either A, B, C or D from left to right. The first test involved no
treatment (control group), which determined the behavior of D. melanogaster without any stimulant. D.
melanogaster was transferred into the apparatus by attaching the vial to the apparatus. After the transfer,
the apparatus was plugged with a stopper. Then, after 2, 6 and 10 minutes, the number of D. melanogaster
in each quadrant were counted and recorded. After 10 minutes, D. melanogaster were transferred back to
the vials via funnel. Then, the study proceeded by testing the behavior of D. melanogaster in the presence
of phototaxis (light), thermotaxis (heat), chemotaxis (apple cider vinegar and caraway) and gravitaxis
(angle of apparatus). Each tests tested the significance of the alternative hypothesis. If D. melanogaster
moved toward the treatment, it responded positively to it, but if D. melanogaster moved away from the
treatment, it responded negatively to it.
The phototaxis test consisted of applying LED light directly to quadrant D with aluminum to
block the light from overhead. For the thermotaxis test, a heating pad was placed under quadrant D only.
In the apple cider vinegar test, ten drops of ACV were placed onto a circular pad located at the end of the
apparatus, near quadrant D. Next, the caraway treatment test consisted of placing two drops of caraway
oil onto circular pad that was located at the end of the apparatus (quadrant D). The final treatment tested
gravitaxis (angle of the apparatus), which the apparatus was tilted at a 30 degrees angle from the lab
bench with having quadrant D at the top. Lastly, a class pool data was generated for the 10 minute results,
and the Chi-square test was performed to determine the statistical significance of each test with different
treatments. Once the chi-square values were calculated, it was used to determine the p value based on the
degrees of freedom.
3. Results
The location of D. melanogaster was counted in 2, 6 and 10 minutes after D. melanogaster were
transferred into the apparatus from the vial. In Figure 1, the class pooled data revealed that at 10 minutes,
the apparatus with no treatment had 336 D. melanogaster, and of those 336 D. melanogaster, there were
63 in quadrant A, 79 in B, 123 in C and 71 in D. The phototaxis test had 143 D. melanogaster in total
with 8 in quadrant A, 36 in B, 74 in C and 25 in D. And the thermotaxis test had 32 D. melanogaster in
quadrant A, 37 in B, 25 in C and 18 in D for a total of 112. The apparatus that was testing chemotaxis
(apple cider vinegar) had 30 D. melanogaster in quadrant A, 37 in B, 35 in C and 19 in D for a total of
121. The apparatus that was testing chemotaxis (caraway) had 190 D. melanogaster with 41 in quadrant
A, 75 in B, 64 in C and 10 in D. Lastly, the apparatus that was testing gravitaxis had 181 D. melanogaster
in total with 143 in quadrant A, 22 in B, 8 in C and 8 in D (Figure 1).
The chi-squared statistical test was performed to calculate the chi-squared values and to
determine the p values, which concluded the statistical significance of the tests. For the phototaxis

Olivia Liang - Animal Behavior Lab Report

treatment, the chi-squared value was 23.22 with a p value less than 0.01 (p<0.01). And for the thermotaxis
treatment, the chi-squared value was 17.69 with p<0.01. Also, the experiment that dealt with apple cider
vinegar as chemotaxis, the chi-squared values was 8.57, which resulted in a p value that is less than 0.05,
but greater than 0.01. On the other hand, with caraway as chemotaxis, the chi-squared value was 44.49,
which resulted in a lower p value of less than 0.01. Finally, the gravitaxis treatment had the highest chisquared value of 435.56 and a p value of less than 0.01.
Based on the chi-squared statistical test, the p values are all less than 0.05 (p < 0.05), which
means that there is statistical significance on the behavior of D. melanogaster in response to different
types of treatments, such as phototaxis (light), thermotaxis (heat), chemotaxis (apple cider vinegar),
chemotaxis (caraway) and gravitaxis (angle of the apparatus).

Treatment's Impact on the Behavior of D. melanogaster

Figure 1: Class pooled data - The number of D. melanogaster that were recorded in each quadrant for
each treatment after 10 minutes.

Chi-Squared Statistical Test on Different Treatments

Olivia Liang - Animal Behavior Lab Report

Treatment

Chi-Squared Value (X^2)

P Value

Phototaxis

23.22

p < 0.01

Thermotaxis

17.69

p < 0.01

Chemotaxis - ACV

8.57

0.05 > p > 0.01

Chemotaxis - Caraway

44.49

p < 0.01

Gravitaxis

435.56

p < 0.01

Table 1: The statistical significance of each treatment based on chi-squared statistical test of
independence, which concluded that all treatments are significant to influence the behavior of D.
melanogaster.
4. Discussion
The study tests the research question, how will different treatments, such as light, heat, apple
cider vinegar, caraway and the angle of the apparatus affect the number of D. melanogaster in each
section (A, B, C and D) of the tube? The studys purpose was to identify behavioral features and patterns
of D. melanogaster that allowed researchers to conclude the complexity of human biology through the
study of this model organism.
The alternative hypothesis stated that the differences in treatments will have an impact on the
behavior of D. melanogaster and its location inside of the apparatus, which predicted that the presence of
stimuli, such as phototaxis (light), thermotaxis (heat), chemotaxis (apple cider vinegar), chemotaxis
(caraway) and gravitaxis (angle of the apparatus) will cause D. melanogaster to move away from the
treatment. The hypothesis and prediction were supported based off of statistical data, which conveyed that
in the presence of stimuli, D. melanogaster responded negatively to the stimuli by moving away from the
treatment. Additionally, based on the chi-squared statistical test, the p values are all less than 0.05 (Table
1), which means that there is statistical significance on the behavior of D. melanogaster in response to
different types of treatments. Also, because phototaxis, thermotaxis, chemotaxis (caraway) and gravitaxis
had a p value of less than 0.01 (Table 1), the distribution was even less likely to be chance-driven. As a
result, the null hypothesis was rejected. Vang, Medvedev and Adlers research article supported the
concluded results of this study (Vang et. al 2012).
To further explain the behavior of D. melanogaster, a potential follow-up study is to perform an
outdoor experiment to see if sunlight will influence the behavior of D. melanogaster while keeping the
treatments and apparatus the same. The laboratorys non-natural light may cause inactivity or overactivity

Olivia Liang - Animal Behavior Lab Report

in D. melanogaster; however, the study of D. melanogaster behavior under natural sunlight will verify the
results and determine if the type of environment or sunlight will affect the behavior of D. melanogaster.
References
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Sexual Receptivity. Proceedings of the Royal Society B: Biological Sciences, 273(1584), 315
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