Chapters 1 5 References and Appendices

HUA SIONG COLLEGE OF ILOILO
SENIOR HIGH SCHOOL

Iloilo City 1
˹ Mutagenic Effects Of Different Powdered Juices To The Phenotypes Of Fruit Flies

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(Drosophila melanogaster)
Milan Mil Grace T. Aragon
Sharina D. Huang
Niel Frankie T. Lim
Kai Gabrielle R. Sanico
Chapter 1
Introduction of the Study
Chapter 1 covers seven parts: (1) Background of the Study, (2) Statement of the
Problem, (3) Research Hypotheses, (4) Paradigm and Conceptual Framework of the Study,
(5) Significance of the Study, (6) Scope and Delimitations of the Study, (7) Definition of
Terms.
Part One, Background of the Study, presents the overview of the study.
Part Two, Statement of the Problems and Hypotheses, presents the general and
specific problems alongside with the hypotheses of the study.
Part Three, Research Hypotheses, provides several statements as predictions of
the study’s outcome in relation to the given research questions.
Part Four, Paradigm, illustrates the relationship of the dependent and independent
variables of the study.
Part Five, Significance of the Study, enumerates and explains the benefits that
may be derived from the findings of the study.
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Part Six, Scope and Delimitation of the Study, specifies the scope and limits of the
study.
Part Seven, Definition of Terms, presents the conceptual and operational
meanings of important terms used in the study.
Background of the Study
For an organism to continue thriving in its environment, English naturalist and
biologist Charles Robert Darwin introduced evolution – a theory that explains how species
undergo a descent of modification throughout time which enables them to produce new
species and “share a common ancestor” (Khan Academy, n.d.). The mechanism of this
concept, according to Andaya et al. (2015), is supported by Natural Selection. This
fundamental process towards evolution emphasizes the significance of variation of genetic
information for a specific species’ existence on Earth; it highlights the importance of
passing of heritable traits that favor survival and reproduction of species towards their
offspring in order for them to adapt to their unstable environment.
One of the factors that can lead to evolution is through mutation, which is
described by the American Phytopathological Society (2018) as the “change in the DNA at
a particular locus of an organism.” It plays a significant role in evolution and is the
wellspring of genetic variation. A mutation in species could be caused by the following:
ultraviolet light, radiation, and some chemicals (Andaya et al., 2015). Although a first step
in evolution as it changes the DNA sequence of a particular gene to create new alleles
(American Phytopatholoigical Society, 2018), mutation does not guarantee to prolong the
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existence of a certain species in Earth; in fact, it could be the very reason of their
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extinction.
The U.S. National Library of Medicine in 2015 explained that each cell of an
organism heavily relies on numerous proteins to do their particular roles in the body
precisely in accordance to the place they are located and time to function. A slight
alteration to one of these proteins due to gene mutations can prevent them from working
properly, critically impairing the body to normally develop and acquire medical conditions.
Mutated cells can be directly acquired from the cataclysms of mutation or inherited from
parents leading to monogenetic disorders and multifactorial inheritance disorders,
respectively (National Human Genome Research Institute, 2015). Diseases belonging to
monogenetic disorders are sickle cell diseases which are prevalent among of African
descent, whereas most cancers and heart disease are under the category of multifactorial
inheritance disorders.
Acknowledging the severity of this problem, it becomes most imperative for the
general public to be aware of the possible mutagenic effects brought by their powdered
juices. A report by Adams (2017) suggested that most consumers turn to powdered drink
mixes because of its low price and convenience. This supports the statement made by
Kantar Worldpanel in 2013, wherein their study disclosed that the on-the-go lifestyle of
many Filipinos has made them constantly search for fast and easy food and beverage
choices. Conolly (2018) also reported that according to the Journal of AOAC International
in 2006, these beverages are designed to contain supplemental quantities of vitamins (e.g.
C, B-2, B-5) and minerals.
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˹ Despite its aim to provide supplemental quantities of nutrients to consumers,

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powdered juices contain a high amount of fructose corn syrup, in which the amount is
almost comparable to soft drinks (Elliot, 2016). Groves (2018) described fructose as a
monosaccharide that is “commonly found in vegetable and fruits and added to processed
food in the form of high-fructose corn syrup.” Before the body can use fructose for energy,
the human liver has to convert it into glucose. The excess fructose in the human body is
usually turned into cholesterol and triglycerides – leading to insulin resistance, deposition
of liver, and impaired body lipids that may result to negative health effects such as obesity,
fatty liver disease, and high cholesterol, respectively (Gunnars, 2018). With that in mind,
the researchers were prompted to examine the mutagenic effects of powdered juices
through subjecting it to fruit flies (Drosophila melanogaster), which are cheap yet reliable
experimental sources in the field of mutation.
The use of fruit flies (Drosophila melanogaster) in laboratory experiments was
pioneered by American zoologist and geneticist Thomas Hunt Morgan, which led him to
develop a hypothesis of sex-linked characters. Alongside with C.B. Bridges, A.H.
Sturtevant, and H.J. Muller, Morgan was able to explain and publish The Mechanism of
Mendelian Heredity after developing his work on fruit flies (Drosophila melanogaster) into
a large-scale theory of heredity. Morgan received the Nobel Prize in 1933 for his discovery
of Drosophila’s hereditary transmission mechanism (Allen, 2017). As of now, fruit flies
(Drosophila melanogaster) have remained to be a widely used subject in research
involving genetics since Nichols and Pandey (2011) affirmed in their study that common
fruit flies (Drosophila melanogaster) are highly tractable genetic models because 75% of
human disease-causing genes are credited to function similarly in the fruit flies’
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(Drosophila melanogaster) biological, physiological and neurological attributes. This, in
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turn, makes it a reliable model organism in this study.
Statement of Problem
Generally, this study was conducted to examine the mutagenic effects of different
powdered juices on the phenotypes of fruit flies (Drosophila melanogaster).
Specifically, this study aimed to answer the following questions:
1. What is the mutation that occurred in the fruit flies (Drosophila melanogaster)?
2. Is there a significant difference in the eye color of the fruit flies (Drosophila
melanogaster) when subjected to different powdered juices as mutagen?
3. Is there a significant difference in the body color of the fruit flies (Drosophila
melanogaster) when subjected to different powdered juices as mutagen?
4. Is there a significant difference in the wing structure of the fruit flies
(Drosophila melanogaster) when subjected to different powdered juices as
mutagen?
5. Is there a significant difference in the head formation of the fruit flies
(Drosophila melanogaster) when subjected to different powdered juices as
mutagen?
6. Is there a significant relationship between the sex of the fruit flies (Drosophila
melanogaster) and the mutation?
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Research Hypotheses
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In view of the aforementioned problems, the following hypotheses were
advanced:
1. There is no significant difference in the eye color of the fruit flies (Drosophila
melanogaster) that were subjected to different powdered juices as mutagens.
2. There is no significant difference in the body color of the fruit flies (Drosophila
melanogaster) that were subjected to different powdered juices as mutagens.
3. There is no significant difference in the wing structure of the fruit flies
(Drosophila melanogaster) that were subjected to different powdered juices
as mutagens.
4. There is no significant difference in the head formation of the fruit flies
(Drosophila melanogaster) that were subjected to different powdered juices
as mutagens.
5. There is no significant relationship between the sex of the fruit flies
(Drosophila melanogaster) and mutation.
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Research Paradigm
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Eye Color
(Phenotype)
Powdered Drink
Mixes (Mutagen)
Body Color
1. Orange (Phenotype)
2. Iced Tea
3. Pineapple
Sex of the Flies Wing Structure

(Phenotype)
1. Female
2. Male
Head Formation
(Phenotype)
Independent Variable Dependent Variable
Figure 1. The paradigm of the study.
Figure 1 shows the paradigm of the study. The researchers classified sexes and
powdered drink mixes as the independent variables of the study. Powdered drink mixes
were identified to be the mutagen as adding it to the experimental set-up may lead to
changes of physical characteristics of the test subjects, which are the fruit flies
(Drosophila melanogaster). These powdered beverages may be classified into three: (1)
Orange, (2) Iced Tea, and (3) Pineapple. On the other hand, the phenotypes of the fruit
flies (Drosophila melanogaster), namely the eye color, body color, wing structure, and
head formation, were considered as the dependent variables of the study. The changes in
their physical characteristics are present even after their given period of stayed in the
cultured environment.
This paradigm is supported by the Nutritional Epigenetic Theory. As stated by
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Landecker (2011), the Nutritional Epigenetic theory seeks to explain the effects of
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nutrition on gene expression. The surroundings of an organism can affect how its genes
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reveal characteristics and it can all be done without changing the genetic code itself. DNA
methylation and stable chromatin modification mediates epigenetic phenomena in animals
and plants. The scientific community has shown significant interest, whether external
factors modify the establishment and maintenance of epigenetic modifications, and could
thereby affect gene expression and phenotype. The thoughts or views that this research
explains also highlight the connections and the contribution of the subject's ancestral
environmental experiences.
Significance of the Study
The findings, which were revealed by the data, may benefit certain groups and
the benefits they may be able to gain are as follows:
Education. This study can give students and teachers, especially in the Science,
Technology, Engineering, and Mathematics strand, a deeper information or higher skill in
their field regarding genetics and inheritance.
Fast Food Industries. The results gathered in this study can help fast food
industries assess better alternatives in beverages for their customers’ health.
Future Researchers. The information gathered may serve as basis or as
supporting foundation for other researches. The researchers can also use the findings of
the study to check the validity of their work or improve this study by adding or
experimenting more variables or sample size.
Genetics. Especially in the field of mutation, researchers can acquire additional
knowledge on how different powdered juices can affect the genes of fruit flies (Drosophila
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melanogaster) – which was given us a broader scope on how mutation works in all
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species.
Government. This study can help the government implement protocols on
powdered beverage industries to ensure the safety and health of the general public.
Powdered Beverage Industries. Since powdered juice is mostly used in their
products such as beverages or food, food manufacturers can acquire additional
knowledge regarding the mutagenic effects of powdered juice. With that said, this study
could be the basis of the concentrations of powdered juice applied to their products.
Students. The students from the STEM strand can gain knowledge of how fruit
flies (Drosophila melanogaster) undergo mutation when subjected to a specific mutagen.
Other than that, they have an edge of how to conduct this type of experiment and may
yield better results.
Teachers. Teachers from the STEM strand can acquire insights on how to assess
this type of write-ups and experiments regarding the mutation of fruit flies (Drosophila
melanogaster) when subjected to a specific mutagen and may help their students perform
this activity better.
Scope and Delimitations
This paper examined the mutagenic effects of powdered fruit juice on fruit flies
(Drosophila melanogaster), especially in the following phenotypes: eye color, body color,
wing structure, and head formation. It did not ensure that all fruit flies to be observed
were wild. The study also revealed if there is a significant difference in the aforementioned
phenotypes when subjected to the selected mutagen.
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˹ Since the experiment usually lasts for a week or two, the researchers conducted
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this in their homes and brought it to their school for microscopic observation of the
subjects. The researchers compared twenty (20) fruit flies (Drosophila melanogaster)
from the controlled set-up and replicated it thrice to three (3) experimental set-ups that
contains twenty (20) fruit flies (Drosophila melanogaster) per set-up that were replicated
thrice as well. The data gathered and observed from the microscope were analyzed and
synthesized through the statistical tool called IBM-SPSS Statistics 22.
The study was carried out from February 14 – March 5, 2019. The experiment was
done in the laboratory of the Hua Siong College of Iloilo – Main Campus.
Definition of Terms
Banana. This is a tropical plant that is well-known to be a good source of
Potassium. If unripe, the color of this fruit is green and yellow when it has matured (Durr,
2010).
In this study, the banana refers to a fruit used to attract the common fruit flies
(Drosophila melanogaster) for the research. This also serves as one of the set-ups used in
the study.
Chloroform. This is a liquid with a scent designed to make people or any creature
subjected to it unconscious when they smell it (Cambridge Dictionary, 2018).
In this study, it is used as an instrument which was subjected to the fruit flies
(Drosophila melanogaster) in order for the observation under the microscope be
conducted.
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˹ Fruit Flies (Drosophila melanogaster). These are easy and inexpensive

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laboratory pets that served as a versatile model in biomedical researches (Jennings, 2011).
These creatures can be genetically modified in many ways.
In this study, this term refers to the subjects experimented exposed to the
selected mutagen.
Mutagen. Any cataclysm of mutation that is capable of remodelling the cell’s
genetic material through altering the DNA’s structure (Encyclopaedia Brittanica, 2018).
In this study, the term refers to the different powdered drink mixes added to the
banana as experimental set up which were used to trigger the mutation of common fruit
flies (Drosophila melanogaster).
Mutation. A process in which the cell of prokaryote or eukaryote undergoes an
alteration in the genetic material (Griffith, 2017). It can be passed off to the organism’s
descendants.
In this study, mutation refers to the change in the eye color, body color, wing
structure and head formation that occurred in the fruit flies (Drosophila melanogaster).
Powdered Drink Mixes. These are beverages in powdered form that are
formulated to correspond to its liquid counterparts (Sutherland and Varnam, 2012).
In this study, powdered drink mixes were served as the mutagen that were
subjected to fruit flies (Drosophila melanogaster). It is also the independent variable of
the study.
Sex. It is the animals’ set of biological attributes that is associated with physical
and physiological features (Canadian Institutes of Health Research, 2015).
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In this study, the sex of the fruit flies (Drosophila melanogaster) served as one of
the independent variables in the study.
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˹ Chapter 2
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Review of Related Literature
This chapter reviews some concepts and research literature which has significant
bearings and relevance on the present study. The purpose of this section is to provide
general information about research studies that have been conducted and are related to
the present investigation.
Chapter 2 covers seven parts: (1) Conceptual Literature, (2) Related Studies, (3)
Synthesis, (4) Theoretical Framework, (5) Conceptual Paradigm, (6) Research Hypotheses,
and (7) Definition of Terms.
Part One, Conceptual Literature, tackles the key concepts of the study.
Part Two, Related Studies, summarizes the major findings of previous studies
Part Three, Theoretical Framework, discusses the theories, Natural Selection and
Sex-Linked Chromosomes, where the study is anchored on.
Part Four, Synthesis, combines the studies presented in the previous section and
draws conclusions from them, as well as relating them to the current study while
accentuating the research gaps.
Conceptual Literature
Nutritional Epigenetics
This study relies on the concept of nutritional epigenetics, which was studied by
Hannah Landecker in a research study entitled “Food as exposure: Nutritional epigenetics
and the new metabolism” last 2011. Nutritional epigenetics, for social science, is a
subtopic of life science whose analysis reveals a concentrated form of a wider shift in the
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understanding of the relationship of food and metabolism. Instead of the classic food to
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energy and body matter metabolism, food can now affect the genome and the physiology
of the organism's body. The researcher believed that nutrients consumed in the prenatal
and early postnatal stages of others’ life impacts early adult diseases. Exposure to food at
a young age can help build immunity later on in the long run.
Landecker’s research analyzes how food has become environment or a subtopic in
nutritional epigenetics, focusing on the experimental formalization of food. Rodent models
were mirror human life in this experiment generated a concept of food as a form of
molecular exposure. This scientific discourse has profound implications for how food is
perceived, manufactured and regulated, as well as for social theories and analyses of the
social body that have a long history of imbrication with scientific models of metabolism.
Mutation in Fruit Flies
An online article published by Exploratorium Exhibit explains that mutated fruit
flies can have different changes in their wing structure, eye color, body color, and head
formation, depending on the gene that was defected. To illustrate, a normal fruit fly is
described to have wings, black and tan striped bodies, and bright red eyes with normal
antennas sticking out. Alterations in the wing structure can produce short-winged flies and
curly-winged flies, wherein short-winged flies have a defect in both of its vestigial gene on
the second chromosome, impairing their ability to fly; whereas, only one copy of the
second chromosome of the curly-winged flies were altered. On the other hand, yellow and
ebony fruit flies exhibit abnormal body color. Yellow flies have a defect in their X
chromosome, while ebony ones have a defect in their third chromosomes. Odd colors in
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the eyes of fruit flies occur if there is a defect in the white gene, which normally produces
red pigment in the eyes. For instance, the orange-eyed fruit flies’ white gene only works
partially, thereby producing fewer red pigments. The white gene of white-eyed flies is
wholly defective, which produces no red pigments at all. In the head formation of fruit flies,
mutations could either lead to eyeless flies or leg-headed flies. Eyeless flies have a defect
in their “eyes absent” gene. The previously mentioned gene is in charge of instructing cells
in the larvae to form eyes. Conversely, the leg-headed flies have a defect in the
“antennapedia” gene, which is in charge of instructing some somatic cells to become legs.
In their case, the antennapedia gene wrongly instructs the antenna to become legs.
Drosophila melanogaster in experiments
The use of Drosophila melanogaster in experiments concerning genetics or
mutation is supported by Echalier’s “Drosophila Cells in Culture (Second Edition)” in 2018.
Echalier states that Drosophila melanogaster offers a balance of simplicity and complexity
with its DNA content, compared to that of Escherichia coli, is 50 times greater and when
compared to that of mammals, it is 30 times smaller. It has been used in various
researches for more than a hundred years and offers many advantages for experimental
studies. Functional and comparative genomic studies revealed a high degree of gene
conservation between flies and other species. In experiments, the use of cultured cells
offers a wide number of advantages. Many Drosophila cultured cell lines, as well as media
conditions, have been established, facilitating a wealth of studies.
Nichols and Pandey’s “Human Disease in Drosophila melanogaster and the role of
Fly in Therapeutic Drug Discovery” in 2011 also affirmed Echalier’s claim of fruit flies’
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being in gene-related experiments. In understanding the molecular mechanism of the
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human diseases, the common fruit flies are considered to be a highly tractable genetic
model because 75% of human disease-causing genes are credited to function similarly in
the fruit flies’ biological, physiological, and neurological attribute. Small molecules based
primarily in vitro cell culture and enzymatic or receptor binding assays were subjected to
high-throughput screening during the traditional approach in the therapeutic discovery
process. Most of the positive effects in vitro screens were deemed to be toxic and
ineffective in subsequent validations of experiment towards whole-animal models –
leading to urge researchers to search for new tools and platforms in the discovery area to
overcome the limitations. The fruit flies holds a great promise in discovering for an
enhanced rate of discovery of higher quality leads. Powerful genetics, highly conserved
disease pathways, and very low comparative costs are some of their several unique
features as models of human diseases. Other than that, they can be used from low to
high-throughput drug screens. In this study, Nichols and Pandey (2011) reviewed the
fruit fly’s basic biology, discussed human disease models and the possible opportunities
that may lead to therapeutic discoveries of central nervous system and inflammatory
disorders, cardiovascular disease, cancer, and diabetes. The researchers also provided
sources for people interested in making fly as their research model in human disease and
using Drosophila melanogaster in the drug discovery process.
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Related Studies
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Fruit Flies as Model in Experiments
Bellen, Wangler, and Yamamoto’s “Fruit Flies in Biomedical Research” in 2015
reported that many scientists complained and experienced financial problems in managing
their equipment in research experiments. There was a general decline in fund support for
the scientific research from the National Science Foundation. The Drosophila
melanogaster was suggested to be used as a cheaper instrument in experiments
pertaining to biological process and genetic material. It became a better concept for
modelling that has the translational impact for genetic disease and other medical
implication.
The claim of Bellen, Wangler, and Yamamoto were affirmed in Jenning’s
“Drosophila - a versatile model in biology and medicine” last 2013. The study states that
the fruit fly Drosophila melanogaster has been used in biomedical research across century
to study a wide range of phenomena. A few of the technical advantages of utilizing
Drosophila melanogaster than other vertebrate models are the following: they are cheap
and not hard to culture in the laboratory, have a shorter life cycle, they propagate large
numbers of externally laid embryos, and can be genetically modified in many ways.
Studies concerning Drosophila melanogaster have made advancements in understanding
regenerative biology and can give large contributions to the regenerative medicines in the
future.
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Process of Making Powdered Beverages
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The “Preparation of Beverages Powder from Fruits” of Hla and Khaing in 2011
described the process behind preparing fruit juices in powder form. Powdered beverages
have a storage stability that does not change in time. It can preserve organoleptic
properties such as flavor, taste, and aroma. It can be transported easily due to its volume
and size. Hla and Khaing’s study emphasized the preparation of the fruit juice in powder
form with the aim of retaining its natural flavor and aroma when it is rehydrated. The
powders were characterized by its physical and chemical properties such as its acidity, pH,
ash content, soluble solid content, total solid content, moisture content, specific gravity,
and protein content in order to determine and differentiate the quality of other branded
products in Myanmar’s local markets. It was revealed that storage temperature plays a
pivotal role in dried juices. The researchers believed that the processes involved in making
dried powder should be done more meticulously in order to lower fruit powder’s moisture
and to reduce its hygroscopic nature. Fiber content should also be measured for the
preparedness of the fruit powder.
Properties of Powdered Juices and its Feasibility as Mutagen
The research paper of Bezerra et. al. (2016) entitled “Cytotoxic and genotoxic
potential of powdered juices” explains that the inexpensive cost, availability in different
fruit flavors, and minimal effort exerted in preparing beverages makes powdered juices
more favorable than industrialized beverages; thus, being widely consumed by the
general consensus. These products contain several kinds of food additives resulting in its
complex formulation. Since no scientific studies were conducted in determining the
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toxicity of these products, Bezerra et. al. evaluated the toxicity at the cellular level of
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industrialized powdered juices of orange and guava flavors of three different food
companies. The researchers analyzed this through the use of meristem cells of Allium
cepa L. at the exposure times of 24 and 48 hours, and two concentrations, 30 g/1000 mL
which are considered ideal for consumption according to the label of the products, and 30
g/500 mL was made. Both flavors of juices from the three companies in both
concentrations and the two exposure times developed significant antiproliferative effect to
root meristem cells and generated a statistically substantial number of mitotic spindle
changes and micronuclei in cells in the test system used. This study concluded that all the
samples of powdered juices under this research’s studied conditions displayed cytotoxic
and genotoxic potential.
Physical and chemical properties of pineapple juice, agglomerate and drink
For the physical and chemical properties of pineapple juice, agglomerate, Lee
(2005) explains that the process of fluidized bed drying and agglomeration can convert
pineapple juice into powder or agglomerate. Ground sugar was selected as the carrier by
the researcher while other necessary minor additives such as anhydrous citric acid,
pineapple flavor and permitted food color were abled to strengthen the flavor, sugar-acid
balance, and make the product appealing to the eyes. The process temperature for the
optimum operation variable used was 40 °C, 20 m3/ha as the volumetric airflow rate
which resulted in an air velocity of 1.5 m/s, an atomization pressure of 3 bar and a pump
flow rate of 6 g/min. The binder which was used for the agglomeration process within the
fluidized bed dryer that was sprayed at the start of the drying process was fresh pineapple
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juice. The juice recovery was 43.82% and the moisture content of the fruit was
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comparatively high at 88.36%. The juice that was strained was light yellow in color with
L* = 44.44, a* = 1.14, b* = 25.43 color value, total titratable acidity (TTA) = 0.47%, pH
= 3.75, total soluble solids (TSS) = 9.5 and viscosity = 2.55 cP. The size distribution of the
agglomerate’s particles exhibited that less than 10% of the particles was maintained on
the large mesh no. (mesh nos. 20, 30, 40 and 45). Approximately 15% of the particle size
was between 250-300 μm and 45% of the particles was finer than 250 μm. The pineapple
agglomerate has a color value of L* = 90.57, a* = –0.51, b* = 25.93, 0.67 g/ml was the
bulk density and 3.1% was the moisture content. The pineapple drink was correlated to
the fresh pineapple juice which had colour values of L* = 64.62, a* = 0.53, b* = 25.90,
the TTA was 0.40%, the pH was 2.88 and the TSS was 9.0. The viscosity was slightly more
viscous than the juice at 2.77 cP. The sensory evaluation’s results revealed that there are
no significant differences in the color, flavor, sweetness, sourness, and overall
acceptability of the drink and juice; however, all of the study’s attributes acquired higher
scores given to the juice except for the overall acceptability which gave the same score for
both the drink and juice. It was also emphasized that due to the sugar used as the carrier,
the result of the pineapple agglomerate’s chemical analysis showed that the total content
of sugar and carbohydrate and energy values were high. The amount of the other
elements were considered comparatively low except for the content of potassium for the
pineapple agglomerate in comparison to the orange flavored drink powder which contains
high content of calcium, sodium, potassium and vitamin C. This can be interpreted by the
fact that fortification of the orange flavoured drink powder added to increase the
amount of these contents.
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Genotoxicity Analysis of Pineapple Extract on Oral Cancer Cell Line by DNA
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Fragmentation
Gayathri, Mahtani, and Priya (2016) aimed to evaluate the genotoxicity of the
pineapple extract on oral cancer cell line using the DNA fragmentation with the purpose of
determining the genotoxicity’s effect towards oral cancer cells through DNA fragmentation.
Genotoxicity was defined to be the chemical substances’ property of destroying genetic
information – leading to mutation that might possibly develop into cancer. Bromelain is a
mixture of substances that can be acquired from the pineapple stem and core. A group of
protein-digesting enzymes called cysteine proteinases can also be acquired from the
pineapple crude extract, which contains Vitamin C. The oral cancer cell lines were
maintained as Bromelain was added to it, which was then followed by the DNA isolation
and DNA fragmentation. After the Agarose gel electrophoresis, the DNA was observed
using ethidium bromide under the Ultraviolet light. The results of this study revealed that
DNA fragmentation occurred in all the three concentrations. This study has proven that
Bromelain contain anti-cancerous properties.
Effect of thermal treatments on antioxidant and antiradical activity of blood
orange juice
Blood orange juice is known for its richness in phenolic compound and its
antioxidant properties. Iannoccari et. al. (2003) subjected orange segments to thermal
treatments before it produced juice to evaluate the changes in antioxidant and chemical
composition. The thermal treatment involved in this study increased the main phenolic
substances of orange juices such as anthocyanins and total cinnamates whereas it
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decreased the ascorbic acid. Results showed that the antioxidants properties increased in
thermally-treated samples when evaluated in a lipoxygenase-inocleic acid system. On the
other hand, free radical scavenging activity was enhanced in untreated juices when
evaluated through ESR spin trapping of hydroxyl radical and DPPH quenching. The study
inferred that there is a possible relationship between the changes in composition and the
changes in antioxidant and antiradical properties.
Influence of orange juice of the genotoxicity induced by alkylating agents: An
in vivo analysis
Erdtmann et. al (2005) suggested in their research study that there is an
association between diets rich in fresh fruit and vegetables and a decreased incidence of
cancers as supported by the substantial epidemiological evidences. Alkylating agents such
as Methyl methanesulfonate (MMS) and cyclophosphamide (CP) differ in the mode of their
actin as Methyl methanesulfonate (MMS) is a directly-acting or monofunctional agent
whereas cyclophosphamide (CP) is a bifunctional agent which obligates metabolic
activation to a reactive metabolite. Prior to and after the treatment with orange juice, the
mice were treated orally using the Methyl methanesulfonate (MMS) and
cyclophosphamide (CP) in order to evaluate whether the orange juice lessened the
damage in the DNA which was induced by the alkylating agents. The comet assay was
used to evaluate the DNA damage in the peripheral white blood cells. These experimental
conditions indicated that orange juice reduced the extent of DNA damage caused by the
mutagens. For Methyl methanesulfonate (MMS), the antigenotoxic effect of orange juice
was both protective (orange juice pre-treatment) and reparative (orange juice
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post-treatment). On the other hand, in cyclophosphamide (CP), the effect was only
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reparative. The components of orange juice exhibit several biological effects, including
modulating metabolization/detoxification routes and acting as targets of toxicants. As the
two drugs differ in mechanisms, it also suggests that both also differ in protective effects.
The ability of the in vivo comet assay to detect in vivo modulation of Methyl
methanesulfonate (MMS) and cyclophosphamide (CP) mutagenicity by the orange juice
was demonstrated in the results.
Potential Dangers of Instant Iced-Tea by Kimberly Stakal (2011)
Lacking of nutritional supplements and antioxidants in instant iced tea brings
potential dangers. Stakal (2011) reported that many researchers have discovered that
instant iced tea have other mixes unlike the freshly brewed teas as they contain only little
to no antioxidants in their chemical makeup. It was also revealed that these commercial
drinks contain things that should not be there, such as fluoride, artificial sweeteners, and
BHA.
During the preparation of instant iced tea, they are exposed to low pressure before
being freeze dried and finally powdered. The USDA discovered that more than 90 to 100
per cent of the original antioxidants content was lost during the production of instant iced
tea. Stakal also cited that the study of Journal of Agriculture and Food Chemistry revealed
that pre-bottled iced green tea preparations are devoid of the nutritive polyphenols called
catechins.
School of Medicine researchers has disclosed that there are dangerously high
levels of fluoride in instant iced tea. The EPA sets the limit of fluoride consumption at
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4ppm but some commercial iced tea contains 6.5 ppm upwards. Back in 2005, this news
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spread all over the nation, as a middle-aged woman who suffered from spine pain due
to hyper-dense bones because of commercial iced tea.
There are also a number of additives present in instant iced tea ranging from
artificial sweeteners to artificial dyes to preservatives. BHA is an additive that is used to
“preserve freshness” in the food and it is linked to all types of organ toxicity. Several
studies conducted by the National Institute of Health showed the correlation between BHA
and liver, thyroid and kidney problems in lab tests. Additionally, the Convention for the
Protection of the Marine Environment of the North-East Atlantic listed BHA as a chemical
of potential concern because of its toxicity and ability to bioaccumulate in the
environment.
Theoretical Framework
The renowned theory of evolution, proposed by the English naturalist and biologist
Charles Darwin in 1859, is where the present study is anchored on. It seeks to describe
how heritable physical or behavioral traits of a certain organism are modified throughout
time. The mechanism behind this is through natural selection, which has wealthy
evidences of its validity coming from the following scientific disciplines: paleontology,
geology, genetics, and developmental biology (Than, 2018).
Aside from Darwin’s theory, this study heavily relies on Thomas Hunt Morgan’s
experiment in fruit flies (Drosophila melanogaster) – which developed the hypothesis of
sex-linked characters. As Morgan observed thousands of fruit flies in his experiment, one
white-eyed fruit fly was obtained. Morgan then began to breed the white-eyed fruit fly and
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noticed that the trait was only present in males of one generation of fruit flies. He
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continued breeding the flies discovered “the genetic factor controlling the eye color in flies
was on the same chromosome that determined sex” (Gleason, 2017). With the results,
eye color and sex were both tied to chromosomes, which led to the establishment that
chromosomes carry the genes that allow traits to be inherited by the offsprings.
Morgan studied the Drosophila melanogaster in which he identified as mutants, or
individual flies that has its heritable characteristics. In May 1910, he observed a single
white eye male fly that was classified as a mutant out of the thousand fruit flies as both
male and female flies usually possess red eyes. It was inferred that the fruit fly’s abnormal
eye color resulted from recessive mutation in the genetic information. The mutant fly was
bred to pure-red female fruit flies in order for Morgan to determine how mutation was
inherited through successive generations. It was revealed in the results that female fruit
flies from the second generation have red eyes. To check if it followed Mendel’s
inheritance patterns, Morgan mated the second generation red-eyed female fruit flies to
the original white-eyed mutant males. There should be one white-eyed fly in every three
red-eyed flies, regardless of sex, in the resulting generation. Although the expected ratio
of white-eyed to red-eyed fruit flies was observed, Morgan concluded that Mendel’s
inheritance pattern did not equally distribute its population across males and females as
most of the white-eyed flies were male.
After the aforementioned experiment, Morgan hypothesized that the controlling
eye color is located on the X chromosomes. Female flies have two X chromosomes, while
males have one X chromosome and one Y chromosome. X-linked is a trait that correlates
with the factor of X chromosomes. All of the X-linked traits were inherited by male flies
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because they only have one X chromosome. Female flies, on the other hand, need
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X-linked trait in both X chromosomes. In his breeding experiment, the first generation of
flies contained only males with white eyes because of the controlling eye color on the X
chromosome. Male flies inherit white eyes because it was present on their X chromosome.
Female flies do not inherit white eyes because it is only present on one of their X
chromosomes.
To test if the white-eyed trait was on the X chromosome, Morgan mated other
group of fruit flies and predicted what were be the ratios of sex and eye color if his
hypothesis were true. He compared the experiment’s results with his predicted results and
determined his hypothesis is supported only by one mating. Morgan mated a red-eyed
male and white-eyed female fruit fly. He observed and predicted that half of the flies
would be red-eyed female and the other half would be white-eyed male. The results
showed that only male flies were capable to inherit the white eyed trait because they only
have one X chromosome.
Sex-Linked Recessive
As explained by the US National Library of Medicine in 2018, sex-linked diseases are
passed from generation to generation through one of the sex chromosomes: X and Y
chromosomes. Dominant inheritance happens when a gene from one parent carries a
disease-causing gene that dominates the normal gene from the other parent. On the other
hand, recessive inheritance occurs when both parents have abnormal genes. The condition
of the disease may be mild or non-existent if only one parent is the carrier of such disease.
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A carrier is referred to someone with one abnormal gene but does not exhibit the
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symptoms of the disease; he or she can pass the abnormal gene to his or her children
Sex-linked recessive is often referred to as X-linked. As males only have one X
chromosome, X-linked recessive diseases mostly occur in males. A single recessive gene on
the X chromosome could cause the disease. Since the Y chromosome, which is the other
half of the XY gene pair in males, does not contain most genes of the X chromosome, males
are not protected from the sex-linked diseases. Some diseases which occurred from a
recessive gene on the X chromosome are hemophilia and Duchenne muscular dystrophy.
If the child’s mother is a carrier of a certain disease while his father is not, there is
a 25% chance that the child will be a healthy boy, a boy with the disease, a healthy girl, or
a girl with a disease. If only the father is the carrier, the outcomes would either be a
healthy boy or a carrier girl without a disease.
Females can also be affected with an X-linked disorder, but the occurrence is very
rare. Since females have two X chromosomes, they will be affected by the disease if both
X chromosome from each parent carries an abnormal gene. This could occur in the two
scenarios below.
For instance, if the mother is a carrier and the father suffers from the disease, there
is a 25% possibility that their child would be a healthy boy, a boy suffering from the disease,
a healthy girl, and a girl suffering from the disease. If both parents suffer from the disease,
their children will also inherit their disease regardless of their sex.
As these two scenarios rarely occur, X-linked recessive diseases are sometimes
referred to as a male only disease. However, this is not technically correct. The “skewed
X-inactivation” occurs when female carriers can have a normal X chromosome that is
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abnormally inactivated. Symptoms similar to males or mild symptoms may be experienced
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by these females.
Synthesis
In the literatures presented above, Drosophila melanogaster has proven to be an
efficient model in studying genetics or mutation as explained by Nichols and Pandey in
2011, Jennings in 2013, and Echalier in 2018; however, there were plenty of gaps present
in the study’s independent variable, which is the different powdered drink mix.
Powdered beverages are favored by most people because of its storage stability,
ability to preserve organoleptic properties longer, inexpensive cost, and convenience in
preparing (Hla and Khaing, 2011; Bezerra et. al., 2016). Even so, the properties of
powdered juices were revealed to have cytotoxic and genotoxic potential in the research
of Bezerra et. al. Other scholarly journals have also disclosed its adverse health effects to
human health like in the report of Stakal regarding instant iced tea. Stakal (2011)
explained that during the preparation of instant iced tea, 90 to 100 per cent of the original
antioxidants of the content was lost. Moreover, it contains dangerous levels of fluoride
and BHA. Meanwhile, Lee (2005) founded out that pineapple agglomerate has high sugar
content, carbohydrate content, and energy values. Lastly, the study of Iannaccori in 2003
showed that the orange segments of blood orange juice indicated a decrease in ascorbic
acid.
Other than the study regarding the properties of pineapple and orange juice
properties to be limited and outdated, it becomes most noticeable that the previous
studies did not determine the powdered beverages’ mutagenicity despite being able to
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reveal harmful contents of a specific powdered juice and its cytotoxicity and genotoxicity.
It must be understood that not all genotoxic substances are mutagenic. The absence
thereof prompted the researchers to fill in the gaps by testing the mutagenic effects of
different powdered juices to Drosophila melanogaster.
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˹ Chapter 3
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Research Design and Methodology
Chapter 3 consists of three parts: (1) Research Design, (2) Materials and
Procedures, and (3) Data Analysis Procedure.
Part One, Research Design, restates the reason for conducting the study, identifies
the tools that are used, and justifies the research paradigm of the study.
Part Two, Materials and Procedures, describes the instruments used and the steps
in gathering the data.
Part Three, Data Analysis, discusses the procedures for data analysis.
Research Design
This study was conducted to determine the mutagenic effects brought by different
powdered juices to the common fruit flies. In a more particular manner, this study
intended to examine the changes on the following phenotypes of the research subjects: (1)
eye color, (2) body color, (3) wing structure, and (4) head formation. In addition, the
relationship between sex and mutation was also investigated.
The design used in this study was experimental research, which is under the
breadth of quantitative research. Barrot (2017) explained that the experimental research
aims to “establish a cause-effect relationship” and “applies variable manipulation more
extensively.” He further elaborated that this type of design helps ensure that the
researchers exhibit greater control of their subjects and provides less threat to the study’s
validity. Posttest Only Design was employed by the researchers to determine the
intervention’s (X) effects to the group of subjects (fruit flies) that were introduced to it and
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to compare it to the controlled set-up through a microscopic examination. The subjects,
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sharing the same characteristics were randomly assigned (RA) to the controlled group and
three (3) experimental groups. All of the set-ups had three replications to ensure the
experiment’s reliability.
Control Group Posttest
Experimental Group X Posttest

RA 1
2
3
Figure 2. Posttest Design of the study.
Materials
The materials used in the study were categorized under the following processes:
Luring of Common Fruit Flies, Preparation of Control and Experimental Set-ups,
Microscopic Observation and Data Gathering and Data Analysis.
Luring of Common Fruit Flies
For the luring of common fruit flies, the researchers gathered at least twelve (12)
bananas from one stem, twelve (12) rubber bands, six (6) used foot stockings, thirteen
(13) plastic jars of the same area and volume, and a spoon.
Preparation of Control and Experimental Set-ups
After the fruit fly collection set-up, the researchers prepared the following
materials for the controlled and experimental set-ups in the school laboratory: twelve (12)
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bananas from one stem, another twelve (12) bananas from one stem weighing 44.5
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grams for the experiment proper, thirteen (13) plastic jars from the fruit fly collection
set-up, twelve (12) used stockings, twelve (12) rubber bands, three (3) different
powdered juices weighing 4.45 grams each, twelve (12) watch glasses, twelve (12) plastic
basins, mortar, pestle, water, a triple beam balance, and a plastic bag. The researchers
wore their personal laboratory gowns, gloves, and masks for sanitary purposes.
Microscopic Observation of the First Filial Generation
Three (3) dissecting microscopes, three (3) watch glasses, three (3) pairs of
tweezers, at least three (3) microscope slides, beaker, dropper, chloroform, cotton balls,
and four (4) check sheets were used by the researchers during their first and second
microscopic observation of the specimens. For sanitary purposes, all of the researchers
wore their respective laboratory gowns, gloves, and masks during the observation of their
subjects.
Data Analysis
Finally, the check sheets that were served as their data from the microscopic
observation and laptops with an SPSS program were used in determining the results of
this experiment.
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Procedures
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The procedures that were conducted for the experiment were the following:
•Collection of Common Fruit Flies

1
•Preparation of Experimental and Controlled Set-ups

2
•Release of Parental Generation

3
•Experiment Proper
4
•Microscopic Observation of the Fruit Flies

5
Figure 3. Procedures of the experiment
Fruit Fly Collection
After the researchers acquired the needed materials, they found the best location
in attracting the common fruit flies which is at 342 Rizal Street, Lapuz, Iloilo City. Once the
place was settled, the researchers put six (6) of the plastic jars in the area where the fruit
flies are most accessible. The one in charge then lightly mashed the bananas with the help
of a spoon and assigned two (2) of the bananas with their skins in each jar as these
bananas were served as the baits for the subjects. The researchers left them for at least
3-5 days until the desirable amount of fruit flies was achieved in every jar. They then
covered the jars with foot stockings and secured them with rubber bands before
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transferring them to the school laboratory.
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Experimental and Controlled Set-Ups
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This section is sub-divided into three: (1) Preparation, (2) Release of Parental
Generation, and (3) Experiment Proper.
Preparation
The researchers neatly arranged the six (6) plastic jars filled with fruit flies and six
(6) empty jars in a spacious area within the laboratory. After that, the researchers
removed the rubber band of the plastic jar that is filled with fruit flies and placed one (1)
empty plastic jar above it. They made sure that both jars were sticking close to each other.
The researchers swiftly removed the foot stocking of the filled jar and waited for a few
minutes until a sufficient amount of fruit flies has transferred to the empty jar. To finally
separate them, the researchers lightly raised the upper jar in a miniscule distance and
they quickly covered the lower jar with foot stocking and the upper jar with a plastic bag
simultaneously. They then carefully placed the upper jar in the working area and covered
it with a foot stocking quickly after removing the plastic bag. After that, the researchers
peeled a banana and lightly mashed them with the aid of a mortar and pestle and placed
their skin and flesh inside the empty jar by swiftly uncovering the foot stocking and
covering them again. This was done in a quick manner to avoid many fruit flies from
escaping. While the previous step was being executed, the researchers removed the old
bananas from the filled jar by slightly uncovering the foot stocking and replaced them with
a newly peeled banana without forgetting to quickly cover the jar after it was done. They
then tightened both of the jars’ lids with rubber bands. They repeated the previous steps
until all jars that were secured with a foot stocking and rubber bands had a desirable
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amount of fruit flies with a lightly mashed banana for them to feed on. To avoid ants from
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swarming the jars, the researchers filled the twelve (12) plastic basins with a small
amount of water and placed one jar on each basin. They left these set-ups for at least 3-5
days for the fruit flies to reproduce.
Release of Parental Generation
Once there was an adequate supply of maggots in each jar, the researchers
released the parental generation of fruit flies by transferring them outside the school
laboratory momentarily. They carefully removed the foot stockings in each jar so that all
of the adult fruit flies can escape. After a few minutes of ensuring that only the maggots or
the first filial generation were left, they covered the jars with foot stockings and brought
them back in the school laboratory.
Experiment Proper
The researchers prepared the different powdered juices, twelve (12) bananas,
triple beam balance, and other materials needed in the working station. They put one (1)
banana in the triple beam balance to determine its mass. Since all of the bananas that
were subjected to the set-ups must have the same mass, the researchers decided on what
should be the bananas’ uniformed mass. In this case, the researchers agreed that all of
the bananas must weigh 44.5 grams. A spatula may be needed to adjust the mass of
bananas. After achieving the banana’s desired mass with the aid of the triple beam
balance, the researchers placed it in a watch glass. This was done repeatedly until twelve
(12) of the bananas were done. For the powdered juices that served as the mutagens, its
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mass should be one-tenth of the banana which is 4.45 grams. The researchers placed a
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thin piece of paper and put the powdered juice on the triple beam balance to adjust its
mass to 4.45 grams. Once the desired mass of the mutagen is achieved, they placed them
in one of the bananas. They did this thrice for each powdered juice. After nine (9) of the
bananas were subjected to the mutagen, the student researchers labelled the jars with
the A being the controlled set-ups and B, C, D as the experimental set-ups. They took out
the foot stocking of one jar and removed the old banana placed there. The researchers
replaced them with the banana in accordance with their label. They covered it with the
foot stocking again. The researchers repeated the last three procedures until all of the
set-ups were finished. They neatly arranged them in a Randomized Complete Block
Design and waited for at least 5-7 days for the filial generation to become adult fruit flies.
The researchers utilized the Randomized Complete Block Design. Grant (2018)
explained that this design divides the subjects into subgroups called blocks in order to
determine the differences of different treatments and to minimize the variability among
the units within groups (blocks). Therefore, within each block, the conditions were as
homogeneous as possible. It can have any number of treatments and blocks. In this
study’s case, there were about four treatments with three replications each. Each block
was designed to contain at least twenty specimens for microscopic observation.
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Block 1 Block 2 Block 3
A1 B2 C3
C1 D2 A3
D1 A2 B3
B1 C2 D3
Figure 4. Four (4) treatments with three (3) replications arranged in a Randomized
Complete Block Design (RCBD)
Microscopic Observation
The researchers assembled three (3) dissecting microscopes with its microscope
glass in the working station. They brought the thirteenth empty jar alongside with a set-up
that was to be observed. They then placed the empty jar above the set-up and quickly
removed the foot stockings of the jar filled with the fruit flies. After a desirable amount of
fruit flies had gone up to the empty jar, the researchers simultaneously covered the upper
jar with plastic and the lower jar with foot stocking. They placed the upper jar in the
working station and carefully remove the plastic. Chloroform was then poured into the
beaker and two (2) cotton balls were placed in a watch glass. With the aid of a dropper,
the researchers released five (5) drops of chloroform in each cotton ball and swiftly
inserted it to the set-up. They waited until all of the fruit flies were unconscious. At least
five (5) unconscious fruit flies were then transferred into the microscope slides and they
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were observed in terms of their phenotypes in the dissecting microscope. The cotton ball
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on the watch glass that was subjected with chloroform was displayed near the observed
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specimens to keep them from moving. The researchers called out the characteristics of
each fruit fly’s phenotype (eye color, body color, wing structure, and head formation) and
sex so that the other researcher in charge of the checklist can determine whether the
specimen exhibits a mutagenic effect on its phenotypes and can input the data accurately.
After the specimen had been observed, the researchers placed them in a watch glass and
disposed all of the specimens in that set-up were observed. They repeated the previous
procedures for another set-up until all twelve (12) of them were finished. The researchers
then transferred all of the set-ups outside the laboratory and released the first filial
generation.
Data Analysis Procedure
For the analyses of the data, version 22 of the leading statistical software acquired
by IBM in 2009 named SPSS Statistics was utilized. Data was inputted in the following
labelled columns: fruit flies’ number, fruit flies with or without mutagen, change in eye
color, change in body color, change in wing structure, change in head formation, and sex.
Variables were all nominal. Fruit flies with or without mutagen served as the independent
variables alongside its sex. Meanwhile, the change in eye color, body color, wing structure,
and head formation of fruit flies were treated as dependent variables. The researchers
used One-Way ANOVA, a parametric test that was designed to assess significant
differences between three (3) or more set-ups (Laerd, 2018). This test was used in
determining whether there is a significant difference in the eye color, body color, wing
structure, and head formation of the research subjects. They further employed the
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Fisher’s least significant difference – a test “determining the smallest significance between
the two means” so that the researchers can generate direct comparisons between two (2)
set-ups since One-Way ANOVA cannot tell which set-up differed from another (Glen,
2014). Pearson’s r, which Laerd (2018) described as a parametric test, was designed to
“measure the strength and direction” of two variables. This test was used to answer
whether there is a significant relationship between the sex of the fruit flies and the
mutation.
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˹ Chapter 4
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Results and Discussion
Chapter 4 covers three parts: (1) Introductory Paragraph, (2) Descriptive Findings,
and (3) Inferential Findings.
Part One, Introductory Paragraph, provides a restatement of the research
questions or objectives of the study.
Part Two, Descriptive Findings, presents the descriptive data and their relevant
analyses and discussion.
Part Three, Inferential Findings, presents results of statistical analyses and
discussion.
Introductory Paragraph
The present study intended to examine the mutation that occurred in the fruit flies
when subjected to different powdered juices. It specifically aimed to determine if there is
a significant relationship between the sex of the fruit flies and the mutation, and the
significant difference in the following phenotypes: (1) eye color, (2) body color, (3) wing
structure, and (4) head formation of the fruit flies.
Descriptive Findings
The total quantity of fruit flies was 240 from twelve (12) different containers. Each
container consisted of twenty (20) fruit flies with different mutagen. The mean
represented the percentage population of fruit flies that mutated or were affected by the
mutagen. The phenotype that received the most number of mutations was the wing
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structure with the mean of 0.57 or 57% population of the fruit fly. After the wing structure
was the head formation, which had a mean of 0.51 or 51% population of the fruit fly. It
was followed by the eye color of the fruit flies that mutated with the mean of 0.38 or 38%
of the population. The body color attained to be the least number of mutations with the
mean of 0.16 or 16% of the fruit flies’ population.
Table 1
Descriptive Statistics of the Mutations in Phenotypes
N Mean Std. Deviation
EYE COLOR 240 0.38 0.49
BODY COLOR 240 0.16 0.37
WING STRUCTURE 240 0.57 0.50
HEAD FORMATION 240 0.51 0.50
Total 0.40 0.46
Inferential Findings
Difference in Eye Color of the Fruit Flies
Table 2 shows the Analysis of Variance of the difference between the eye color of
the fruit flies. ANOVA revealed that there is a significant difference (F = 100.001, p=
0.000) in the eye color of the fruit flies that were subjected to different powdered juices as
mutagens as the p value is lower than the alpha level; thus, the researchers rejected their
first null hypothesis.
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Table 2
˺
Analysis of Variance of the difference in the eye color of the fruit flies when subjected to
different mutagens
N Mean Std. Deviation
EYE COLOR 240 0.38 0.49
BODY COLOR 240 0.16 0.37
WING STRUCTURE 240 0.57 0.50
HEAD FORMATION 240 0.51 0.50
Total 0.40 0.46
Table 3 Post Hoc Analysis (Fisher’s LSD) further revealed that there was indeed a
significant difference between control and iced tea (MD = -.81667*, p = .000) in favor of
the iced tea; whereas, the pineapple juice also showed a significant difference when
compared to the control group and with pineapple juice inducing more mutations (MD =
-.33333*, p = .000). Iced tea and pineapple juice both displayed a significant difference
when compared to the orange juice in favor to the iced tea (MD = -.93333*, p = .000) and
pineapple juice (MD = -.45000*, p = .000). Between the set-ups of iced tea and pineapple
which both exhibited a significant difference, iced tea was able to induce more mutations
(MD = -.48333*, p = .000) in the eye color of the fruit flies. A mutation occurred in the
control set-up because the researchers did not ensure that all of the fruit flies were wild.
This table implied that iced tea has the greatest influence in altering the eye color of the
fruit flies compared to the other mutagens.
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Table 3
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Post hoc analysis (Fisher’s LSD) of the difference in the eye color of fruit flies when
subjected to different mutagens
MUTAGEN Mean Difference Sig.
CONTROL
ICED TEA -.81667* .000
PINEAPPLE JUICE -.33333* .000

ORANGE JUICE
ICED TEA -.93333* .000

ICED TEA
PINEAPPLE JUICE .48333* .000
Although the study of Bezerra et. al. (2016) tested the cytotoxicity and genotixicity
of powdered juices to a plant’s phenotypes, the findings of the research can be directly
linked to previous study as all cytotoxicity, genotoxicity, and mutagenicity may lead to a
cell’s abnormal development. Furthermore, it affirms the report of Stakal (2011) that
instant iced tea indeed brings adverse effects to human health alongside with Lee’s study
(2005) regarding pineapple agglomerate and Iannaccori’s (2003) research on orange
segments from blood orange juice. It may have suggested that instant iced tea’s
dangerous levels of fluorine and BHA, pineapple agglomerate’s high sugar and
carbohydrate content, and orange juice’s decreased in ascorbic acid are what prompted
the defect of the fruit flies’ white gene – which normally produces red pigments in their
eyes. A defect in the white gene may result to the specimens having orange or white eyes
instead of red.
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Difference in Body Color of the Fruit Flies
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Table 4 shows the Analysis of Variance of the difference between the body color of
the fruit flies. ANOVA revealed that there is no significant difference (F = 1.670, p= 0.000)
in the body color of the fruit flies that were subjected to different powdered juices as
mutagens as the p value is lower than the alpha level; thus, the researchers retained their
second null hypothesis.
Table 4
Analysis of Variance of the difference in the body color of the fruit flies when subjected to
different mutagens
Sum of
df Mean Square F Sig.
Squares
Between
.679 3 .226 1.670 .174
Groups
Within
31.983 236 .136
Groups
Total 32.663 239
Unlike with the eye-color results, these findings cannot be directly linked the study
of Bezerraet. al. (2016) which tested the cytotoxicity and genotixicity of powdered juices
carry to a plant’s phenotypes as there is no significant mutagenicity displayed by the fruit
flies’ body color. It does not necessarily refute the reports of Iannaccori, Lee, and Stakal
that orange juice, pineapple agglomerate, and orange segment from juice may potentially
threaten human health; but it does imply decreased in ascorbic acid of orange juice,
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dangerous levels of fluorine and BHA of instant iced tea, and high sugar and carbohydrate
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˹
content of pineapple agglomerate are not enough to alter fruit flies’ “yellow gene” and
˺
“ebony gene” – which are located the X chromosome and third chromosome, respectively.
A defect in the “yellow” gene may result to the specimen’s body being wholly yellow
whereas a defect in the “ebony” gene leads the black pigments to accumulate all over the
fruit fly’s body the normal body color of fruit flies is described to be a black-and tan
striped.
Difference in Wing Structure of the Fruit Flies
Table 5 shows the Analysis of Variance of the difference between wing structure of
the fruit flies. ANOVA revealed that there is a significant difference (F = 100.001, p=
0.000) in the wing structure of the fruit flies that were subjected to different powdered
juices as mutagens as the p value is lower than the alpha level; thus, the researchers
rejected their third null hypothesis.
Table 5
Analysis of Variance of the difference in the eye color of the fruit flies when subjected to
different mutagens
Sum of Mean
df F Sig.
Squares Square
Between
7.333 3 2.444 11.180 .000
Groups
Within Groups 51.600 236 .219
Total 58.933 239
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˹ Table 6 Post Hoc Analysis (Fisher’s LSD) further revealed that there was indeed a
˺
significant difference between control group and iced tea (MD = -.46667*, p = .000), in
favor of the iced tea. Between the pineapple and the control group which showed a
significant difference when compared, it was pineapple that induced more mutations (MD
= -.23333*, p = .007) as shown in Table 7. Orange and iced tea exhibited a significant
difference in favor to the iced tea (MD = -3.6667*, p = .000). A significant difference was
also displayed between the iced tea and pineapple set-ups wherein it was further proven
by the table below that it was iced tea that induced more mutations than the pineapple
(MD = -.23333*, p = .007) in the wing structure of the fruit flies. A mutation occurred in
the control set-up because the researchers did not ensure that all of the fruit flies were
wild. Akin to the results of the fruit flies’ eye color, iced tea has the greatest influence in
altering the wing structure of the research subjects.
Table 6
Post Hoc analysis (Fisher’s LSD) of the difference in the wing structure of fruit flies when
CONTROL ICED TEA -.46667* .000
ORANGE JUICE ICED TEA -.36667* .000
ICED TEA PINEAPPLE JUICE .23333* .007
With these findings, it can be inferred that the study of Bezerraet. al. in 2016
˻
which tested the powdered juices’ cytotoxicity and genotixicity to a plant’s phenotypes can
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˹
be directly linked to this study’s outcomes as all cytotoxicity, genotoxicity, and
˺
mutagenicity may lead to a cell’s abnormal development. Moreover, it affirms the report of
Stakal (2011) that instant iced tea indeed brings adverse effects to human health
alongside with Lee’s study (2005) regarding pineapple agglomerate and Iannaccori’s
(2003) research on orange segments from blood orange juice and may have suggested
that instant iced tea’s dangerous levels of fluorine and BHA, pineapple agglomerate’s high
sugar and carbohydrate content, and orange juice’s decreased in ascorbic acid are what
prompted the defect of the fruit flies’ “vestigial” gene and “curly” gene located on the
second chromosome. A defect in the “vestigial” gene will lead to short-winged fruit flies,
which is a display of recessive mutations, whereas a defect in the “curly gene”
demonstrates dominant mutation.
Difference of Head Formation of Fruit Flies
Table 7 shows the Analysis of Variance of the difference between head formation
of the fruit flies. ANOVA revealed that there is a significant difference (F = 11.557, p=
0.000) in the head formation of the fruit flies that were subjected to different powdered
juices as mutagens as the p value is lower than the alpha level; thus, the researchers
rejected their fourth null hypothesis.
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˹
Table 7
˺
Analysis of Variance of the difference in the head formation of the fruit flies when subjected to
different mutagens.
Sum of Mean
df F Sig.
Squares Square
Between Groups 7.683 3 2.561 11.557 .000
Within Groups 52.300 236 .222
Total 59.983 239
Table 8 Post Hoc Analysis (Fisher’s LSD) further revealed that there was indeed a
significant difference between control group and iced tea (MD = -.48333*, p = .000), in
favor of the iced tea. A significant difference was also displayed when the iced tea set-ups
were compared to the orange set-ups; it was further proven by the table below that iced
tea induced more mutations (MD = -.36667*, p = .000) than orange. Between the iced
tea and the pineapple set-ups which exhibited a significant difference when compared,
iced tea successfully altered more head formations of the fruit flies than the pineapple
set-ups (MD = -.31667*, p = .000). A mutation occurred in the control set-up because the
researchers did not ensure that all of the fruit flies were wild.
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˹
Table 8
˺
Post Hoc Analysis (Fisher’s LSD) of the difference in the head formation of fruit flies when
CONTROL ICED TEA -.48333* .000
ORANGE JUICE ICED TEA -.36667* .000
ICED TEA PINEAPPLE JUICE .31667* .000
The findings of this study can be directly linked to Bezerra et. al.’s research in 2016
which tested the cytotoxicity and genotixicity of powered juices to a plant’s phenotypes
since all cytotoxicity, genotoxicity, and mutagenicity may lead to a cell’s abnormal
development. It also affirms the report of Stakal (2011) that instant iced tea indeed brings
adverse effects to human health alongside with Lee’s study (2005) regarding pineapple
agglomerate and Iannaccori’s (2003) research on orange segments from blood orange
juice. It may have suggested that instant iced tea’s dangerous levels of fluorine and BHA,
pineapple agglomerate’s high sugar and carbohydrate content, and orange juice’s
decreased in ascorbic acid are what prompted the defect of the fruit flies’ “eyes absent”
gene and “antennapedia” – which normally instructs cells in the larvae to form eyes and
body cells to become legs, respectively. A defect in the “eyes absent” gene may result to
the specimens having no eyes, whereas a defect in the “antennapedia” gene results fruit
flies to have leg-like antennas in their formed head.
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˹
Relationship between Sex and the Mutation
˺
Table 9 revealed that there is no significant relationship (r = 0.000, p = 1.000)
between the fruit flies’ sex and mutation and that the association between the two was
negligible.
The results of this study supported the concept of Sex-Linked Recessive that both
parents should be the carrier of such disease in order their offspring to possibly inherit and
suffer from the disease, regardless of their sex. It also affirmed the claim of the US National
Library of Medicine in 2018 that it is technically incorrect to refer X-linked recessive
diseases as a male only disease.
Table 9
Pearson’s r correlation of coefficient in the relationship of sex of the fruit flies and mutation
SEX MUTATION
SEX Pearson Correlation 1 .000
Sig. (2-tailed) 1.000
N 240 240
MUTATION Pearson Correlation .000 1
Sig. (2-tailed) 1.000
N 240 240
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˹ Chapter 5
˺
Conclusions and Recommendations
Chapter 5 consists of three parts: (1) Summary, (2) Conclusions, and (3)
Recommendations.
Part One, Summary, summarizes the essential points of the investigation and
findings.
Part Two, Conclusions, presents the inferences drawn from the findings of the
study.
Part Three, Recommendations, offers some recommendations based on the
findings and conclusions drawn from the study.
Summary
Generally, this study was conducted to examine the mutagenic effects of different
powdered juices on fruit flies.
Specifically, this study aims to answer the following questions:
1. What is the mutation that occurred in the fruit flies?
2. Is there a significant difference in the eye color of the fruit flies when subjected
to powdered juices as mutagen?
3. Is there a significant difference in the body color of the fruit flies when
subjected to powdered juices as mutagen?
4. Is there a significant difference in the wing structure of the fruit flies when
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˹ 5. Is there a significant difference in the head formation of the fruit flies when
˺
6. Is there a significant relationship between the sex of the fruit flies and the
mutation?
The researchers compared four (4) different set-ups with each set-up being
replicated thrice. Per block, twenty (20) fruit flies were examined under the dissecting
microscope – having a total of two hundred forty (240) fruit flies being observed.
The data that were simultaneously recorded while observing them was then
analyzed through the statistical tool SPSS. It revealed the following results:
1. There is a significant difference in the eye color of the fruit flies that were
subjected to different powdered drink mixes as mutagens.
2. There is no significant difference in the body color of the fruit flies that were
3. There is a significant difference in the wing structure of the fruit flies that were
4. There is a significant difference in the head formation of the fruit flies that
were subjected to different powdered drink mixes as mutagens.
5. There is no significant relationship between the sex of the fruit flies and
mutation.
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˹
Conclusions
˺
Based on the findings of the study, the researchers concluded that:
1. The researchers concluded that there is a significant difference in the eye
color, wing structure, and head formation of fruit flies as the Iced Tea induced
the most mutations in the aforementioned phenotypes. It is further inferred
that Iced Tea possesses compounds that can trigger mutation in the fruit fly’s
eye color, wing structure, and head formation.
2. Powdered juices such as Orange, Iced tea, and Pineapple juices are
dangerous for the fruit flies to consume. It could be further inferred that
compounds present in the powdered juices can alter the body color of the fruit
flies.
3. A mutation can happen to any fruit flies regardless of its sex.
Recommendations
After the results and conclusions of the study have been posited, this study offers
the following recommendations:
1. Do not to subject Drosophila melanogaster to powdered Iced Tea juice as it is
likely to induce mutations in eye color, wing structure, and head formation. It
is also recommended that humans consume less amount of instant Iced Tea
because 75% of human disease-causing genes are credited to function
similarly on the fruit flies biological, physiological, and neurological attributes.
2. Do not to subject Drosophila melanogaster to powdered Orange, Iced Tea, and
Pineapple juice as it is likely to induce mutations in body color. It is also
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˹ recommended that humans consume less amount of Orange, Iced Tea, and
˺
Pineapple juice because 75% of human disease-causing genes are credited to
function similarly on the fruit flies’ biological, physiological, and neurological
attributes.
3. Future researchers with similar topics can use these findings presented for
support and validation.
4. To be of help to the general public, the scientific community should conduct
several research studies regarding powdered juices and its compounds in
order to inspect if their composition really triggers gene alteration or mutation.
The findings of these studies may improve the health security of powdered
juice consumers.
5. To be of help to the scientific community, the Philippine government should
support and encourage researches concerning this matter. This is because
different powdered juices are widely used by Philippine fast food industries and
widely consumed by many Filipinos; thus, affecting many people. Through
aiding the scientific community in conducting this type of researches, less of
the general consensus will suffer subtle harmful side effects brought by
powdered juices.
6. Lastly and most importantly, the researchers recommend that further studies
should be made wherein its topics aim to explore the mutagenic effects of
different concentrations of different types and hues of powdered juices since
this study only made use of three powdered juices. It will also be better for the
succeeding studies to have wild fruit flies instead of common fruit flies as their
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˹ ˺
subjects so that the results will be more reliable and use a bigger sample in
their experiments by conducting more microscopic observations towards the
second, third, or more filial generations of Drosophila melanogaster.
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˹ ˺
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Andaya, M., Aquino, M, Biong, J., & Valdoz, M. (2015). Science Links: Worktext for
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Bellen, H.J., Wangler, M. F., & Yamamoto, S. (2015). Fruit flies in biomedical
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Bezerra, M.D.S., Malaquias, G.D.S. , Peron, A.P., & Sousa, J.M.D.C. (2016). Cytotoxic and
genotoxic potential of powdered juices. doi.org/10.1590/1678-457X.0006
Canadian Institutes of Health Research. (2015). Definitions of Sex and Gender. Journal of
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Connolly, B. (n.d.). Does Drinking Powdered Drinks Count as Drinking Water?. Retrieved
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˹ https://www.livestrong.com/article/552761-does-drinking-powdered-drinks-count
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-as-drinking-water/
Desiderio, L. (2013, Seotember 9). Rural households driving demand for consumer goods.
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households-driving-demand-consumer-goods
Durr, J. (n.d.). Characteristics of the Banana Fruit. Retrieved from
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Echalier, G. (2017). Drosophila Cells in Culture (Second edition).
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Elliott, B. (2016). 20 Foods With High-Fructose Corn Syrup. Journal of Nutrition. Retrieved
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Exploratorium. (2007). Mutant Fruit Flies. Retrieved from
https://bugscope.beckman.illinois.edu/pdfs/insects/mutant.pdf
Gayathri, R., Mahtani, A.A., & Priya, V. (2016). Genotoxicity Analysis of Pineapple Extract
on Oral Cancer Cell Line by DNA Fragmentation. Retrieved from
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Gleason, K. (2017). “Sex Limited Inheritance in Drosophila” (1910), by Thomas Hunt
Morgan. Embryo Project Encyclopedia. Retrieved from
https://embryo.asu.edu/pages/sex-limited-inheritance-drosophila-1910-thomas-h
unt-morgan
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Griffiths, A.J.F. (2018). Mutation. Retrieved from
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https://www.britannica.com/science/mutation-genetics
Groves, M. (2018). Sucrose vs Glucose vs Fructose: What's the Difference?. Journal of
Nutrition. Retrieved from
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Gunnars, K. (2018). Is Fructose Bad for You? The Surprising Truth. Journal of Nutrition.
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Helmenstine, T. (2019). The Difference Between Control Group and Experimental Group.
Journal of Science. Retrieved from
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Hla, P.K. & Khaing, T.T. (2011). Preparation of Beverages Powder from Fruits. Retrieved
from http://www.myanmar-education.edu.mm/wp
content/uploads/2013/05/27_Pansy_Kyaw_Hla.pdf
Iannoccari, T., Lo Scalzo, R., Morelli, R., Rapisarda, P., & Summa, C., (2004). Effect of
thermal treatments on antioxidant and antiradical activity of blood orange juice.
Journal of Food Chemistry, 85, 41-47. doi.org/10.1016/j.foodchem.2003.05.005
Jennings, B.H. (2011). Drosophila – a versatile model in biology & medicine.
doi.org/10.1016/S1369-7021(11)70113-4
Khan Academy. (2017). Darwin, evolution, & natural selection. Retrieved from
https://www.khanacademy.org/science/biology/her/evolution-and-natural-
selection/a/darwin-evolution-natural-selection
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Laerd Statistics. (n.d.). Spearman’s Rank- Order Correlation. Retrieved from
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https://statistics.laerd.com/statistical-guides/spearmans-rank-order-correlation-st
atistical-guide.php
Landecker, H. (2011). Food as exposure: Nutritional epigenetics and the new metabolism.
Retrieved from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3500842/
Lee, S.Y. (2005). Physical and chemical properties of pineapple juice, agglomerate and
drink. Retrieved from
http://ejtafs.mardi.gov.my/index.php/publication/issues/archive/69-2005/volume
-33-no2/847-330214
National Institutes of Health. (2015). What are Genetic Disorders. Retrieved from
https://www.genome.gov/19016930/faq-about-genetic-disorders/
National Institutes of Health. (2019). What is a gene mutation and how do mutations
occur. Retrieved from
https://ghr.nlm.nih.gov/primer/mutationsanddisorders/genemutation
Nichols, C. & Pandey, U.B. (2011). Human Disease Models in Drosophila
melanogaster and the Role of the Fly in Therapeutic Drug Discovery. Journal of
Pharmacological Reviews, 63, 411-436. doi: 10.1124/pr.110.003293
Stakal, K. (2011). Instant Iced Tea: A Refreshing Glass of Potential Dangers. Journal of
Energetic Health. Retrieved from
https://www.organicauthority.com/health/instant-iced-tea-a-refreshing-glass-of-p
otential-dangers
Than, K. (2018). What is Darwin's Theory of Evolution. History. Retrieved from
https://www.livescience.com/474-controversy-evolution-works.html
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˹ ˺
The Editors of Encyclopaedia Britannica. (2016). Mutagen, Biochemistry. Retrieved from
https://www.britannica.com/science/mutagen
Varnam, A. & Sutherland, J.M. (2012). Beverages: technology, chemistry and
microbiology. Retrieved from
https://books.google.com.ph/books?id=m0vaBwAAQBAJ&pg=PA98&lpg=PA98&d
q=varnam+sutherland+powder+drink+mix&source=bl&ots=x5xbZFF9lK&sig=AC
fU3U05AkZ_knAZgnBz_oUumrISNMgkQ&hl=en&sa=X&ved=2ahUKEwiJ3faeyLPg
AhXIF4gKHdKDC_MQ6AEwB3oECAcQAQ#v=onepage&q=varnam%20sutherland
%20powder%20drink%20mix&f=false
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˹ ˺
APPENDICES
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Iloilo City 63
˹ Appendix A
˺
Communication Letter to Research Adviser
July 18, 2018
JAYSON I. GULA
Faculty Member, High School Department
Hua Siong College of Iloilo – Main Campus
Sir:
Greetings!
In line with partial fulfilment in the subject Practical Research 2, the Grade 12 students in
the Science, Technology, Engineering, and Mathematics (STEM) strand are currently
entitled to conduct a quantitative research related to their field.
We are writing to honourably request your service and expertise to serve as an adviser for
our study. We believe that your knowledge and insights will be valuable and give great
contributions to our research.
Thank you for your consideration and we hope you will be able to fulfill our request.
Respectfully yours,
MILAN MIL GRACE T. ARAGON NIEL FRANKIE T. LIM

Researcher Researcher
SHARINA D. HUANG KAI GABRIELLE R. SANICO

Noted:
PETER PAUL S. LAURENTE
˻ ˼
Research Teacher
SENIOR HIGH SCHOOL
Iloilo City 64
˹ Appendix B
˺
Communication Letter to Statistician
July 18, 2018
ROVELYN C. CALINAWAGAN
Ma’am:
Greetings!
In partial fulfilment of the subject Practical Research 2, the Grade 12 students in the
Science, Technology, Engineering, and Mathematics (STEM) strand are currently entitled
to conduct a quantitative research related to our field.
We are writing to honourably request your service and expertise to serve as a statistician
for our study. We believe that your knowledge and insights will be valuable and give great
Respectfully yours,


Noted:
JAYSON I. GULA
Research Adviser
˻ ˼
SENIOR HIGH SCHOOL
Iloilo City 65
˹ Appendix C
˺
Communication Letter to Research Grammarian
July 18, 2018
APRIL ROSE VALENCIANA

Ma’am:
Greetings!
We are writing to honourably request your service and expertise to serve as a grammarian
for our study. We believe that your knowledge and insights will be valuable and give great
Respectfully yours,


Noted:
JAYSON I. GULA
Research Adviser
˻ ˼
SENIOR HIGH SCHOOL
Iloilo City 66
˹ Appendix D
˺
Communication Letter to Laboratory Assistant
August 9, 2018
FAITH AGUSTIN
Senior High School Laboratory Assistant
Ma’am:
Greetings!
We are writing to honourably request you to allow us in using the school laboratory for our
experimental research.
Respectfully yours,


Noted:
JAYSON I. GULA
Research Adviser
˻ ˼
SENIOR HIGH SCHOOL
Iloilo City 67
˹ Appendix E
˺
Permission Letter for Laboratory Usage
November 19, 2018
WARREN K. UY
Principal, BED
Hua Siong College of Iloilo, Inc.
Sir:
Greetings!
In partial fulfillment in the subject Practical Research 2, the Grade 12 students of the
to conduct a quantitative research related to their field.
We are writing to honorably request for your approval in letting us use the school
laboratory from Novmber 26-December 10, 2018 for our experimental research entitled
“Mutagenic Effects of Different Powdered Juices to Drosophila melanogaster (Fruit Flies)”
as it will help us in our thorough observation of our subjects.
Respectfully yours,


Noted by:
JAYSON I. GULA
Research Adviser
˻ ˼
SENIOR HIGH SCHOOL
Iloilo City 68
˹ ˺
LIGAYA E. LEONARES
Head, Senior High Department
MARC LOUIE A. LIBO-ON

Over-all Head, SHS Department
Approved:
JOSE B. BARCELONA, PH.D.

Assistant Principal, BED
cc: ELY B. BARRERA

AVP for Academic Affair
˻ ˼
SENIOR HIGH SCHOOL
Iloilo City 69
˹ Appendix F
˺
Permission Letter for Laboratory Usage
February 14, 2019
ERIC VINCENT E. CONLU

Head, Administrative Department
Hua Siong College of Iloilo
Sir:
Greetings!
In partial fulfillment in the subject Practical Research 2, the Grade 12 students of the
to conduct a quantitative research related to their field.
We are writing to honorably request for your approval in letting us use the school
laboratory from February 14 – February 28, 2019 for our experimental research entitled
“Mutagenic Effects of Selected Powdered Juices to Drosophila melanogaster (Fruit Flies)”
as it will help us in our thorough observation of our subjects.
Respectfully yours,


Noted by:
JAYSON I. GULA
Research Adviser
˻ ˼
SENIOR HIGH SCHOOL
Iloilo City 70
˹ ˺
LIGAYA E. LEONARES
Head, Senior High Department
MARC LOUIE A. LIBO-ON

Over-all Head, SHS Department
Approved:
JOSE B. BARCELONA, PH.D.

Assistant Principal, BED
cc: ELY B. BARRERA

AVP for Academic Affair
˻ ˼
SENIOR HIGH SCHOOL
Iloilo City 71
˹ Appendix G
˺
Cover Letter for Validator
February 6, 2018
IVAN MON PANES

Dear Sir:
Greetings!
We, the undersigned, are conducting a quantitative research entitled “Mutagenic Effects
of Different Powdered Juices to Drosophila melanogaster (Fruit Flies)” in partial fulfillment
of the requirements for the subject Practical Research 2.
In line with this, we are writing to honorably request for your expertise in validating the
attached experimental procedures which will serve as our instrument for data collection.
Below are the statement of the problems of this study:
1. Is there a significant difference in the eye color of the fruit flies ( Drosophila
melanogaster) that were subjected to powdered juices as mutagen?
2. Is there a significant difference in the body color of the fruit flies ( Drosophila
3. Is there a significant difference in the wing structure of the fruit flies ( Drosophila
4. Is there a significant difference in the formed head of the fruit flies ( Drosophila
Your constructive suggestions in this research instrument will be taken into consideration
and will be highly appreciated. Should you have any questions or clarifications, you may
contact us through our mobile numbers: 09433891713 or 09253561100.
Thank you and we hope for your earliest positive response to this request.
Respectfully yours:
MILAN MIL GRACE ARAGON NIEL FRANKIE LIM

˻ ˼
SENIOR HIGH SCHOOL
Iloilo City 72
˹ ˺
SHARINA HUANG KAI GABRIELLE SANICO
Noted:
JAYSON I. GULA
Research Adviser
˻ ˼
SENIOR HIGH SCHOOL
Iloilo City 73
˹ Appendix H
˺
February 6, 2018
PETER PAUL LAURENTE

Dear Sir:
Greetings!
Respectfully yours:

˻ ˼
SENIOR HIGH SCHOOL
Iloilo City 74
˹ ˺
Noted:
JAYSON I. GULA
Research Adviser
˻ ˼
SENIOR HIGH SCHOOL
Iloilo City 75
˹ Appendix I
˺
February 6, 2018
CLYD OCTAVIANO
Dear Sir:
Greetings!
Respectfully yours:

˻ ˼
SENIOR HIGH SCHOOL
Iloilo City 76
˹ ˺
Noted:
JAYSON I. GULA
Research Adviser
˻ ˼
SENIOR HIGH SCHOOL
Iloilo City 77
˹ Appendix J
˺
Experimental Procedure
Materials:
 Bananas
 Chloroform
 Cotton Balls
 Dissecting Microscope
 Plastic Containers
 Powdered Juices
 Stockings
 Tweezers
 Watch Glass
Fruit Fly Collection Set-up
After acquiring the needed materials in this part, the researchers found the best
location in attracting the wild fruit flies (Drosophila melanogaster) which is at 342 Rizal
Street, Lapuz, Iloilo City.
1. Once the place is settled, put six (6) of the plastic jars in the area where the
fruit flies (Drosophila melanogaster) are most accessible.
2. Lightly mash the bananas with the help of a spoon and assign two (2) of them
with their skins in each jar as they were served as baits for the subjects.
3. Leave them for at least 3-5 days until the desirable amount of fruit flies
(Drosophila melanogaster) is achieved in every jar.
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Iloilo City 78
˹ 4. Cover the jars with foot stockings and secure them with rubber bands before
˺
transferring them to the school laboratory.
Preparation of Experimental and Controlled Set-ups
1. Neatly arrange the six (6) plastic jars filled with fruit flies (Drosophila
melanogaster) and six (6) empty jars in a spacious area within the laboratory.
2. After that, remove the rubber band of a plastic jar that is filled with fruit flies
(Drosophila melanogaster) and place one (1) empty plastic jar above it.
3. Make sure that both jars are sticking close to each other.
4. Swiftly remove the foot stocking of the filled jar and wait for a few minutes
until a sufficient amount of fruit flies (Drosophila melanogaster) has
transferred to the empty jar.
5. To finally separate them, lightly raise the upper jar in a miniscule distance and
quickly cover the lower jar with foot stocking and the upper jar with a plastic
bag simultaneously.
6. Carefully place the upper jar in the working area and cover it with a foot
stocking quickly after removing the plastic bag from above.
7. Peel a banana and lightly mash them with the aid of a mortar and pestle and
place their skin and flesh inside the empty jar by swiftly uncovering the foot
stocking and covering them again.
8. This must be done in a speedy manner to avoid many fruit flies (Drosophila
melanogaster) from escaping the jar.
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˹ 9. While the previous step is being executed, remove the old bananas from the
˺
filled jar by slightly uncovering the foot stocking and replace them with a
newly peeled banana without forgetting to quickly cover jar after it is done.
10. Tighten both of the jars’ grip with rubber bands.
11. Repeat the previous steps until all jars that were secured with a foot stocking
and rubber bands have a desirable amount of fruit flies (Drosophila
melanogaster) with a lightly mashed banana for them to feed on.
12. To avoid ants from swarming the jars, fill the twelve (12) plastic basins with a
small amount of water and place one jar on each basin.
13. Leave these set-ups for at least 3-5 days for the fruit flies (Drosophila
melanogaster) to reproduce.
Release of Parental Generation
1. Once there is an adequate supply of maggots in each jar, release the parental
generation of fruit flies (Drosophila melanogaster) by transferring them
outside the school laboratory momentarily.
2. Carefully remove the foot stockings in each jar so that all of the adult fruit flies
(Drosophila melanogaster) can escape.
3. After a few minutes of ensuring that only the maggots or the first filial
generation were left, cover the jars with foot stockings and bring them back in
the school laboratory.
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˹
Experiment Proper
˺
1. Prepare the different powdered juices, thirteen (13) bananas, triple beam
balance, and other materials needed in the working station.
2. Put one (1) banana in the triple beam balance to determine its mass.
3. Since all of the bananas that were subjected to the set-ups must have the
same mass, decide on what should be the bananas’ uniformed mass.
4. In this case, the researchers agreed that all of the bananas must weigh 44.5
grams.
5. A spatula may be needed to adjust the mass of bananas.
6. After achieving the banana’s desired mass with the aid of the triple beam
balance, place it in a watch glass; do this repeatedly until thirteen (13) of the
bananas were done.
7. For the powdered juices that were served as the mutagen, its mass should be
one-tenth of the banana which is 4.45 grams.
8. Place a thin piece of paper and put the powdered juice on the triple beam
balance to adjust its mass to 4.45 grams.
9. Once the desired mass of the mutagen is achieved, place them in one of the
bananas.
10. Do this thrice for each powdered juice.
11. After nine (9) of the bananas were subjected to the mutagen, label the jars
with the A being the controlled set-ups and B, C, D as the experimental
set-ups.
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˹ 12. Take out the foot stocking of one jar and remove the old banana placed there.
˺
Replace them with the banana in accordance with their label.
13. Cover it with the foot stocking again.
14. Repeat the last three procedures until all of the set-ups is finished.
15. Neatly arrange them in a Randomized Complete Block Design and wait for at
least 5-7 days for the filial generation to become adult fruit flies (Drosophila
melanogaster).
16. The researchers utilized the Randomized Complete Block Design. Grant (2018)
explained that this design divides the subjects into subgroups called blocks in
order to determine the differences of different treatments and to minimize the
variability among the units within groups (blocks).
17. Therefore, within each block, the conditions are as homogeneous as possible.
It can have any number of treatments and blocks.
18. In this study’s case, there are about four treatments with three replications
each.
19. Each block is desired to contain at least twenty specimens for microscopic
observation.
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˹ ˺
Block 1 Block 2 Block 3
A1 B2 C3
C1 D2 A3
D1 A2 B3
B1 C2 D3
Figure 4. Four (4) treatments with three (3) replications arranged in a Randomized
Complete Block Design (RCBD).
Microscopic Observation
1. Assemble three (3) dissecting microscopes with its microscope glass in the
working station.
2. Bring the thirteenth empty jar alongside with a set-up that is to be observed.
Place the empty jar above the set-up and quickly remove the foot stockings of
the jar filled with the fruit flies (Drosophila melanogaster).
3. After a desirable amount of fruit flies (Drosophila melanogaster) has gone up
to the empty jar, simultaneously cover the upper jar with plastic and the lower
jar with foot stocking.
4. Place the upper jar in the working station and carefully remove the plastic.
5. Pour 5mL chloroform into the beaker and place two (2) cotton balls in a watch
glass.
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˹ 6. With the aid of a dropper, release five (5) drops of chloroform in each cotton
˺
ball and swiftly insert it to the set-up.
7. Wait until all of the fruit flies (Drosophila melanogaster) are unconscious.
Transfer at least five (5) unconscious fruit flies (Drosophila melanogaster) into
the microscope slides and start observing their phenotypes in the dissecting
microscope.
8. A cotton on the watch glass that is subjected with chloroform may be
displayed near the observed specimens to keep them from moving.
9. Call out the characteristics of each fruit fly’s phenotype (eye color, body color,
wing structure, and head formation) and sex so that the other researcher in
charge of the checklist can determine whether the specimen exhibits a
mutagenic effect on its phenotypes and can input the data accurately.
10. After the specimen has been observed, place them in a watch glass. Dispose
them when all of the specimens in that set-up were thoroughly observed.
11. Repeat the previous procedures for another set-up until all thirteen (13)
set-ups were finished.
12. Transfer all of the set-ups outside the laboratory and release the first filial
generation.
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Iloilo City 84
˹
Sample Checklist
˺
A1
Put a check (✔) on the corresponding box if the specimen exhibits a mutagenic effect on
its phenotypes, cross (✘) if otherwise.
Head
Fly Number Eye Color Head Color Wing Structure
Formation
1
10
11
12
13
14
15
Figure 5. Checklist for Block A
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˹ Appendix K
˺
Documentation
Set-ups arranged in Randomized Complete Block Design
Application of mutagen to bananas during experiment proper
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˹ The fruit flies inside the plastic container

˺
Preparation of the Controlled and Experimental Set-ups
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Iloilo City 87
˹ Subjecting the specimens to chloroform before microscopic observation

˺
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˹ The microscopic observation of the fruit flies

˺
The actual image of the normal fruit fly
The microscopic view of the mutated fruit fly
˻ ˼

Chapters 1 5 References and Appendices

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HUA SIONG COLLEGE OF ILOILO

SENIOR HIGH SCHOOL

˹ Mutagenic Effects Of Different Powdered Juices To The Phenotypes Of Fruit Flies

Milan Mil Grace T. Aragon

Niel Frankie T. Lim

Kai Gabrielle R. Sanico

Introduction of the Study

specific problems alongside with the hypotheses of the study.

Part Three, Research Hypotheses, provides several statements as predictions of

the study’s outcome in relation to the given research questions.

variables of the study.

may be derived from the findings of the study.

Part Seven, Definition of Terms, presents the conceptual and operational

meanings of important terms used in the study.

Background of the Study

For an organism to continue thriving in its environment, English naturalist and

concept, according to Andaya et al. (2015), is supported by Natural Selection. This

fundamental process towards evolution emphasizes the significance of variation of genetic

information for a specific species’ existence on Earth; it highlights the importance of

offspring in order for them to adapt to their unstable environment.

a particular locus of an organism.” It plays a significant role in evolution and is the

wellspring of genetic variation. A mutation in species could be caused by the following:

parents leading to monogenetic disorders and multifactorial inheritance disorders,

respectively (National Human Genome Research Institute, 2015). Diseases belonging to

C, B-2, B-5) and minerals.

˹ Despite its aim to provide supplemental quantities of nutrients to consumers,

experimental sources in the field of mutation.

The use of fruit flies (Drosophila melanogaster) in laboratory experiments was

develop a hypothesis of sex-linked characters. Alongside with C.B. Bridges, A.H.

of Drosophila’s hereditary transmission mechanism (Allen, 2017). As of now, fruit flies

(Drosophila melanogaster) have remained to be a widely used subject in research

powdered juices on the phenotypes of fruit flies (Drosophila melanogaster).

Specifically, this study aimed to answer the following questions:

melanogaster) when subjected to different powdered juices as mutagen?

melanogaster) when subjected to different powdered juices as mutagen?

4. Is there a significant difference in the wing structure of the fruit flies

(Drosophila melanogaster) when subjected to different powdered juices as

5. Is there a significant difference in the head formation of the fruit flies

(Drosophila melanogaster) when subjected to different powdered juices as

melanogaster) and the mutation?

melanogaster) that were subjected to different powdered juices as mutagens.

melanogaster) that were subjected to different powdered juices as mutagens.

3. There is no significant difference in the wing structure of the fruit flies

(Drosophila melanogaster) that were subjected to different powdered juices

4. There is no significant difference in the head formation of the fruit flies

(Drosophila melanogaster) that were subjected to different powdered juices

5. There is no significant relationship between the sex of the fruit flies

(Drosophila melanogaster) and mutation.

Sex of the Flies Wing Structure

Independent Variable Dependent Variable

Figure 1. The paradigm of the study.

This paradigm is supported by the Nutritional Epigenetic Theory. As stated by

methylation and stable chromatin modification mediates epigenetic phenomena in animals

Significance of the Study

the benefits they may be able to gain are as follows:

Technology, Engineering, and Mathematics strand, a deeper information or higher skill in

their field regarding genetics and inheritance.

industries assess better alternatives in beverages for their customers’ health.

Future Researchers. The information gathered may serve as basis or as

experimenting more variables or sample size.

Genetics. Especially in the field of mutation, researchers can acquire additional

Government. This study can help the government implement protocols on

Powdered Beverage Industries. Since powdered juice is mostly used in their