Chapter 1

The Problem and Its Background

The effective pest control is vital in protecting public health, especially when concerning cockroaches.
Cockroaches are carriers of pathogens that cause their main threats to human health. The American
cockroach (Periplaneta americana) has been identified as a significant public health issue because of
disease transmission including: salmonellosis, dysentery, and even allergies (Oliviera et al., 2023). Due to
the fact that cockroaches breed quickly, and are very adaptive, it is necessary to control cockroach
populations to not create additional public health threats.

The World Health Organization (WHO) suggests that cockroaches should be controlled to limit the
potential of transmitting pathogens (Oliviera et al., 2023). Chemical insecticides have been relied upon as
a remedy in pest control, but the over-reliance on chemical insecticides has created problems, and
health and environmental concerns. Thus, it is beneficial to explore natural and sustainable alternatives
to chemical cockroach pest control that increases dependency upon leather chemicals, while still being
able to reduce pest populations.

The resistance to wide range insecticides has significantly increased alongside the population of
cockroaches, especially in metropolitan areas with more significant populations and inconsistent
sanitation. The World Health Organization (WHO) claims that due to the cockroaches ability to carry
many pathogens, controlling their population is a must if maintaining public health is desired. Although
the use of chemicals in controlling the cockroaches has its long list of risks for both the environment and
human health as recent studies have shown the growing interest in botanical insecticides as a possible
remedy (Ajayi et al., 2023).

In the Philippines, the population of American cockroaches is continuously growing in both urban and
rural areas, in common places where the act of proper waste management and sanitation are hard to
maintain. The danger of cockroach infestation is versatile since they can exist in homes and food
establishments as well, not just in dirty and wet conditions. Several researches regarding the
cockroaches resistance to standard chemical insecticides like pyrethroids, reliance on chemical
insecticides may present difficulties (Ali et al., 2023). The national government has been open about the
possibility of producing organic treatments to cockroach infestation rather than using synthetic
chemicals to control cockroach reproduction.

The American cockroaches are commonly found in kitchens, restrooms and places where waste is kept,
especially in the slums of Metro Manila, including the city of Pasay. The Local Government Units (LGUs)
are constantly looking for safer alternatives in controlling the infestation since the resistance to chemical
insecticides remains a problem. Garlic (Allium sativum) bulbs possess several compounds that have
demonstrated potential insecticidal potential towards limited insects although further studies to
demonstrate efficacy are required as well as there is even less for insects like cockroaches. Garlic is
classified as a minimum-risk pesticide active ingredient by the United States Environmental Protection
Agency under the category of 25B minimum-risk pesticide as it represents little-to-no threat to human
health. Its insecticidal potential makes it a low-cost and environmentally sustainable alternative to other
chemical insecticides. By using garlic products for insect pests we can potentially reduce pest
populations even more while decreasing environmental or health risk (Sciarretta et al., 2022).
The objective of the present study is to evaluate the insecticidal properties of garlic bulbs as botanical
insecticides and the possible implications of such species on American cockroaches. Garlic contains
various elements known to possess antibacterial, antifungal, and insecticidal properties. Research
suggests garlic based treatments could possibly help serve as a botanical insecticidal replacement for
chemicals, particularly water-based products similar to Cyfluthrin. Cockroach infestations can pose a
public health threat at the local, national, and worldwide levels. This study aims to evaluate garlic
extracts as a naturally occurring and sustainable way to possibly help reduce cockroach infestations.

Research Questions

The primary objective of the study is to determine the insecticidal effectiveness of garlic (Allium sativum)
bulb extract as an alternative pesticide used against American cockroaches (Periplaneta americana).

Specifically, The study intends to answer the following questions:

1. What is the observed mortality rate per time interval (in minutes) of the American cockroach
when exposed to the extract?

2. Is there a significant difference between the observed mortality rate per time interval (in
minutes) between (a. positive & negative, b. positive & crude extract, c. negative & crude
extract)?

Hypothesis

1. Ho: Garlic bulb extract does not exhibit an insecticidal effect against American cockroaches

Ha: Garlic bulb extract exhibits an insecticidal effect against American cockroaches.

2. Ho: There is no significant difference in the observed mortality rate per time interval (in minutes)
between;

a. Positive and Negative

b. Positive and Crude extract

c. Negative and Crude extract

Ha: There is a significant difference in observed mortality rate per time interval (in minutes)
between;

a. Positive and Negative

b. Positive and Crude extract

c. Negative and Crude extract

Significance of the Study

Garlic (Allium sativum) bulb extracts have great potential as a natural alternative in controlling pests like
American cockroaches (Periplaneta americana). This study addresses the rising problems of chemical
resistance and the environmental damage created by synthetic insecticides. This research is into using
garlic bulb extract as a plant-based, eco-friendly option and presents alternative pathways for more
sustainable pest controls in homes and farms. This will mean reduced exposure to harmful chemicals
and, by extension, improved public health and safety. Ultimately, this study offers findings into future
policy-making for what could become greener approaches to pest control.

This study offers the following benefits:

Farmers

The study opens avenues for using garlic extracts in pest control programs for farms, a more natural and
environmentally friendly insecticide that would reduce health risks for farmers, prevent soil
contamination, and won’t affect non-target species like birds, bees, and farm animals.

Future Researchers

The research strengthens the foundation for plant-based insecticides as it provides more evidence of
garlic-based insecticides as a reliable alternative for pest control.

Homeowners

Homeowners may benefit from this study, as the research introduces garlic extract as a probably safer
substitute for managing the infestation of the American cockroaches reducing health risks associated
with synthetic chemical insecticides.

Students

This study would be particularly beneficial to students, especially those in Biology, Agriculture, and
Environmental Science circles, as it presents the possibility of using garlic bulb extracts as a natural
insecticide against American cockroaches. It is an important reference study for ecological and
sustainable pest control studies.

General Objective

The study aims to assess the insecticidal properties of the garlic (Allium sativum) bulb extract,
particularly its constituents, and evaluate its potential as a botanical insecticide alternative to synthetic
chemicals in pest control.

1. To determine the mortality rates of cockroaches when exposed to garlic crude extract and assess
its potential to serve as a viable alternative chemical insecticide.

2. To determine the comparison of garlic crude extract with Cyfluthrin in controlling the American
cockroach infestations to evaluate the efficacy of these substances as pest control agents.

3. To determine the effectiveness of garlic crude extract compared to water in controlling American
cockroach infestations and assess whether the garlic bulb is a natural pest control agent.

4. To determine the crude extracts; impact on mortality over a varying duration of time, which will
provide insights into the speed and effectiveness of garlic as a pest control agent.
Conceptual Framework

Table 1.1: IVDV Model

The conceptual framework illustrates the relationship between garlic (Allium sativum) crude extract and
the mortality rate of the American cockroach (Periplaneta americana). The independent variable is garlic
extract, while the dependent variable is mortality rate of the cockroaches. The study aims to evaluate
the botanical insecticidal potential of the garlic extract as a natural alternative to synthetic insecticides.

Data will be gathered based on the mortality rate of cockroaches which is exposed to the crude garlic
extract. Additionally, the efficacy of the crude extract will be compared with both a positive control (a
commercially available insecticide) and a negative control (no treatment or solvent only). Phytochemical
analysis will also be conducted to identify the active constituents which are present in the garlic extract
that may contribute to its insecticidal activity.

Definition of Terms

Different terms are defined herein and to facilitate the understanding of the study.

Crude Extract

The product acquired from the extraction of the bioactive component from fresh garlic bulb using
a solvent that results in a solution or preparation. The extraction method to be used involves crushing
the fresh garlic bulbs to release their active compounds.

Insecticidal Effect

The ability of the molecule drawn out from the garlic bulb causes damage, kill, or change the
behavior of American cockroaches. This can be measured by evaluating the percentage of American
cockroaches that die. At a specified time point, (2 hours and 35 minutes) after treatment with garlic bulb
extract.

Mortality Rate

The percentage of American cockroaches that die from exposure to garlic bulb extract over a given
time. Mortality will be calculated through counting the number of deaths among the cockroaches in
each container to the total number of cockroaches that are exposed to the treatment during a total
observation period of 2 hours and 25 minutes.

Chapter 2

Review of Related Literature and Studies

This chapter includes some of the related literature and studies of both foreign and local literature
conducted on Garlic (Allium sativum) bulb extract and its insecticidal effects on insects, specifically
American cockroach (Periplaneta americana) species. Similar studies conducted by various researchers
are presented to get more data that could contribute to understanding the current research study. These
investigations will enable the researchers to utilize their data and compare it with the outcomes of
experiments. The literature that is relevant to our investigation, both local and foreign, is briefly
reviewed in this chapter.

Insecticidal effects of Garlic (Allium sativum) crude extract

The bulbous plant known as garlic (Allium sativum), which belongs to the Amaryllidaceae family, is
utilized extensively because of its potent flavor and several health advantages. Garlic extracts in both
powder and oil carry active components like amino acid called allicin (Zekeria et al., 2020). These
chemicals are antibiotics that are efficient against a variety of bacterial and fungal species, making this
plant beneficial in food preservation to limit fungal activity. However, allicin does not exist in fresh garlic
bulbs or cloves until it is crushed or cut because an injury to the bulb causes activation to the enzyme
called alliinase, which will metabolize allin to allicin (Onasanya, et al., 2021).
Figure 2.1: Botanical diagram of Garlic (Allium sativum Linnaeus) Plant

Garlic, namely Allium sativum Linnaeus (Asparagales: Amaryllidaceae), has garnered significant interest
because of its strong antibacterial properties. These plants have therapeutic benefits because of certain
bioactive components. The most prominent of these bioactive compounds in garlic are alkaloids, tannins,
flavonoids, phenolic compounds, and organosulfur such as allicin, alliin, diallyl sulfide, diallyl disulfide,
diallyl trisulfide, ajoene and S-allyl-cysteine among many others (Shang et al., 2019, as cited in, El-Saber
Batiha et al., 2020). The repellent activities of different medicinal plants have been reported on
cockroaches (Rejitha et al., 2014, as cited in, Soonwera et al., 2022). Although there aren't many studies
on the impact of garlic on cockroaches' bacterial composition, adding garlic to their diets showed
antibacterial efficacy against several infections. It can therefore be concluded that the inclusions of garlic
at lower concentrations were more effective than when used at higher concentration. (Oladunni et al.,
2023)

Because of the distinctive aroma of Allium sativum it is included in a list of possible plant-based
insecticides, the aroma is described as very pungent while the taste of garlic is said to drive away many
animals and insects, an example is when worms move away because of the aroma of garlic (Hardiansya
et al., 2020). According to Caong et al. (2022) during their experiment, garlic extract yields the most
mortality on cockroaches when they tested four organic insecticides. The organic insecticides they used
in their experiment were Capsicum annuum, Allium cepa, Allium sativum, Nicotiana. The plant garlic
(Allium sativum) contains lectins or chemicals that resemble lectins, like allicin. These may disrupt
various stages of the insect life cycle and give the plant its potent pungency and scent.

Another similar study conducted in 2023 revealed that sulfur compounds from garlic have toxic effects
on various insects. They focused on the toxic effects of organosulfides from garlic using different
concentrations and compared the results with the toxic effects of synthetic chemical insecticides on
different insects. Their study drew the conclusion that the potential of the garlic as an alternative to
some synthetic chemical insecticides and is generally safer for humans and animals and is effective
against pests (Yueyang Liu et al. in 2023)

The biochemical compounds found in garlic bulbs, preparatory procedures of garlic pesticidal products
from fresh and aged bulbs, and the discovery process of bioactive phytochemicals from garlic bulbs. It
also addresses concerns regarding the synthesis of natural insecticides from garlic and explains the use
of garlic products for insecticidal, fungicidal, bactericidal, and nematicidal purposes. Usage of garlic
herbal preparations by growers can reduce non-target exposure to hazardous pesticides and curb
resistance development in pests, thus enabling sustainable organic crop protection. (Samuel & Ariyo,
2022)

According to Rizky et al. (2022), flavonoid compounds themselves function to eradicate insects through
the digestive system or stomach toxins when exposed to insects. They used A. sativum as a botanical
pesticide against Armyworm (Spodoptera litura F.) on lettuce. The active compound content influences
the speed of death. Although caterpillars (such as armyworms) have no direct or close biological
relationship with cockroaches, botanical insecticides like garlic (or garlic-based solutions or powders) can
have some effect on pests alike. Still, they are known as less potent or reliable than synthetic chemical
insecticides based on previous studies.

According to the 2022 book Organic Farming for Sustainable Development by Anjorin Toba Samuel and
Adeniran Lateef Ariyo, the components of garlic could be utilized to create substitutes for traditional
pesticides in the control of agricultural pests and disease-causing pathogens. The naturally occurring
compounds found in garlic bulbs, the techniques for making pesticidal products utilizing aged and fresh
garlic bulbs, and the procedure for identifying bioactive phytochemicals from garlic bulbs are then
elaborated further. Along with discussing concerns regarding the creation of natural pesticides from A.
sativum, it also illustrates how garlic products are used to kill insects, fungi, bacteria, and nematodes.
(Jeyabalan Sangeetha, KAS, 2022)

The primary bioactive ingredient in A. sativum, allicin, accounts for the 70–80% of the sulfur compounds
that give garlic its characteristic smell (Salehi et al., 2019). However, allicin quickly breaks down and
produces other stable organosulfur compounds since it is extremely unstable, reactive, and sensitive to
pH, oxygen, temperature, and organic solvents (Rouf et al., 2020).

These bioactive compounds also give Allium sativum its insecticidal qualities, which work through a
variety of mechanisms such as disrupting cellular functions, interfering with insect respiratory systems,
and discouraging feeding and oviposition (Al-Shuraym LA,2022., Golubkina, et al., 2022). Furthermore, it
is well recognized that A. sativum essential oil and extract targets a number of particular proteins
involved in the metabolism of insects. A. sativum’s efficacy and relative safety for the environment and
organisms that are not targets makes it a suitable organic insecticide. Garlic-based formulations are
generally less hazardous to non-target species than many chemical pesticides, which have a higher risk of
toxicity to the surroundings. Garlic's great biodegradability also guarantees low environmental
persistence, allaying worries about chemical residues and long-term ecological effects (Michel, et al.,
2024).

Garlic may be viewed as an alternative for the currently used organophosphates and carbamates, but it
has a hidden potential risk of increasing toxicity and morbidity to the main organs and neurological
systems of humans. But focusing on the research, Garlic contains an essential oil that can be extracted by
different methods, the best of which is steam distillation. An alternative to organophosphates and
carbamates, the oil derived from garlic has been shown to have insecticidal effects on flying and crawling
insects, particularly cockroaches, houseflies, and mosquitoes. Many previous studies show that garlic oil
effectively kills various insects. (Olakunbi, et al., 2021)

There were notable variations between the treatments and the control utilized in a recent study on the
life cycle of insects. Extensive use of synthetic insecticides in the long term has resulted in residue
accumulation in food products and soil and caused adverse health effects for humans. Therefore,
applying natural insecticides in household and public health sectors is the best alternative to synthetic
insecticides. (Ahmad et al., 2024)

According to Ngegba et al. (2022), Commercially available synthetic pesticides are harmful not only to
people and other animals but also to non-target plants, the environment, and nearby organisms. The
emergence of insect resistance to these artificial pesticides is another issue that has raised worries. This
has led to the growing popularity of biopesticides, which are certain kinds of pesticides made from
natural sources such as bacteria, plants, fungus, animals, and some minerals, as effective alternatives to
conventional pesticides. Biopesticides are environmentally friendly and safer pesticides.

Raw garlic is processed using a variety of techniques in the production of garlic products, including
chopping, smashing, heating, steeping, macerating, lyophilizing, steam distillation, and more. Processing
transforms certain components of raw garlic into other compounds, and processing methods affect the
preparation's chemical and physical characteristics. Both hydrophilic and hydrophobic chemicals are
among its components. Depending on the circumstances and the manner of processing, the components
of raw garlic might transform into different compounds. The aging process produces both hydrophilic
and hydrophobic substances by the enzymatic and chemical reactions, including transformation,
hydrolysis, isomerization, the Maillard reaction, etc. (Kodera et al., 2019)

American Cockroach (Periplaneta americana)

The American cockroach, or Periplaneta americana, is a common urban hygiene pest found in warm,
humid climates worldwide and is strongly associated with human activities. The biggest species of
cockroach is the American cockroach. An adult "waterbug" or American cockroach has a reddish brown
body that is 1.5 to 2 inches long and has a yellowish stripe behind the head. They prefer to live in warm,
dark, moist areas during the day, such as the basements, sewers, and sub-floors. Although they can
survive outdoors at temperatures over 80F, they avoid colder environments. They frequently gather near
boilers, refrigerator motors, hot water pipes, and other heating equipment indoors. There are many
numerous kinds of plants and animals that the American cockroach will consume. Females can live up to
1.5 years, and eggs take 6 to 8 weeks to incubate. Under the right conditions, they quickly grow to
massive proportions (Yap et al., 2021).

The egg, nymph, and adult are the three life stages of this hemimetabolous insect. Whether or not they
mate, females can have fertile offspring as adults, and all of these children are diploid. The American
cockroach frequently produces 12–16 eggs. After being constantly emitted and individually coated in
secretions produced by the colleterial glands, the mature eggs finally develop into an ootheca. The
effective cumulative temperature of eggs is 415.8 ± 38.5 °C days before hatching, with a developmental
threshold temperature of 15.8 °C. Newly born first-instar nymphs must undergo 6–14 molts before
they mature into adults. The American cockroach was shown to have 9–14 nymphal stages when raised
alone at room temperature. A key feature of cockroach life is the adult lifespan, which varies from 29 to
37 degrees Celsius (HI, USA). Hemimetabolous insects often display this intraspecific variation in
nymphal instar numbers, which is influenced by both genetic and environmental factors. The average
longevity of mated adult females was 311 days, whereas the average lifespan of unmated females was
397 days. When the temperature of their environment is at 30 °C, the average longevity of mated adult
females was 187 days, while the average life expectancy of unmated females was 212 days. Data about
the lifespan of the American cockroach may help us understand their species better (Lin et al., 2022).

Figure 2.2: Reproduction of P. americana. (A) An overview of the first oviposition cycle. Red symbol ♀
indicates female adult and ♂ indicate male adult.

Cockroaches are one of the insects that can transmit many illnesses. They are among the most common
bugs aboard trucks, vessels, and aircraft worldwide, especially in areas where food is stored. Chemical
control and environmental management are two methods that can be used to address cockroaches
(Manyullei et al., 2022). A total of 290 (96.6%) of the bacteria collected from cockroaches were
identified. Acinetobacter species, Bacillus species, coagulase-negative Staphylococci (CoNs), Escherichia
coli, Enterococcus species, Enterobacter aerogenes, Klebsiella species, Proteus species, Pseudomonas
aeruginosa, and were among the bacterial species that were isolated. The most prevalent strains of
these were Bacillus, Escherichia coli, and CoNs. (Jalil et al., 2023).

Historically, the use of pesticide sprays has formed the foundation of cockroach control strategies in the
United States. However, in other cases, relying just on insecticides to control cockroaches result in
problems including the emergence of insecticide resistance and cause societal concern because of the
effects on the environment and human health. It is advised to utilize reduced-risk pesticide solutions in
conjunction with other strategies to get around these issues. The use of insecticide baits, traps to
measure infestation levels, cleanliness and hygiene promotion, and public education are all noteworthy
examples of effective cockroach control initiatives (Gondhalekar et al., 2021). Integrated Pest
Management (IPM) practices are essential for long-term, effective treatments for American cockroaches,
especially in light of the growing problem of pesticide resistance. Using only chemical techniques is not
as effective over time as American cockroaches become resistant to conventional pesticides (Damir,
2024).

The study revealed that the most effective repellent for cockroaches was garlic extract. Garlic is
incredibly helpful against cockroaches It emits scents from its leaves and excretions from its roots. These
smells and excretions deter insects, which is why they are considered insect repellent. (Asianparent
(2018); Irina (2019); Srikanth (2018); as cited in Caong et al., 2022)

Throughout history, cockroaches have existed as a disease vector and precursor. An infestation of
cockroaches is a serious home issue. Salmonella is one of the bacteria that cockroaches can carry on
their legs and then transfer to food, where it can lead to food poisoning or infections. The adults must be
kept under control because they are a disease reservoir and are lively and easily spotted wandering
around. Cockroach excrement and body pieces found in household dust can cause asthma attacks and
allergic responses in certain people. Allicin has been demonstrated to possess antiviral, anti-infective,
fungicidal, and bactericidal qualities. Recent research has indicated that garlic is a helpful herbal plant
because it can be used to repel insects that are harmful to some plant pests like cockroaches (Nagal,
2019).

Given how challenging it is to get rid of cockroaches, these insects can deceive people into
believing they are dead. It is uncommon for Periplaneta americana to act as though they are dead unless
there are extreme conditions present. Cockroaches have numerous defense mechanisms to help them
avoid threats. The insect's speed serves as one of their protective mechanisms. Due to their small
stature, they may get away from predators swiftly.

When it comes to feigning to be dead, the majority of insects from the aforementioned species are
excellent actors. They will abruptly fall onto their backs after tucking their small legs into their stomach.
The bug will efficiently hide from the threat by burying its body in an undergrowth and then disappear
after the threat has passed. Cockroaches can still have functioning neurological and muscle systems after
they die. This action causes the roach to twitch and/or move after it dies. Furthermore, the chemicals in
insecticides that are used to kill cockroaches will impact the nervous system, causing loss of coordination
and muscle spasms (Khan, 2024).

Plants and plant-based solutions (essential oil) are safe, affordable, and environmentally beneficial ways
to manage cockroaches. Studies have examined the insecticidal properties of several plant components,
including leaf, stem, seed, flower, and rhizome extract, against cockroaches (Aldo et al., 2021; Rahayu et
al., 2020; as cited in Ukoroije and Bawo, 2019). Plants were used to control two cockroach species,
Periplaneta americana and Blatella germanica, throughout the majority of the research. This research
used Periplaneta americana as the variable in the experiment.

In the Philippines, controlling cockroaches is a crucial component of pest management. Cockroaches,

including common pests like the American cockroach, can multiply quickly and infest your home or place
of work. During their lifetime, cockroaches can have up to 1,000 offspring, making them extremely
resilient pests. Thus, for long-term control, efficient pest management methods are required
(Environment, 2024).
 Chapter 3

Methodology

The methodologies employed in the study are presented in this chapter. The study's design,
sources of data, data collection process, and statistical data treatment are also included.

Research Design

This study utilized a quasi-experimental research design, which does not randomly assign
samples. This study used a quantitative experimental research approach. This study ascertained the
insecticidal efficiency of the extract from garlic (Allium sativum) bulbs against the American Cockroach
(Periplaneta americana). The treatment applied was made from garlic bulb extracts, which were
controlled and measured their insecticidal effects objectively made possible by the quasi-experimental
approach.

Sample Population and Sampling technique

Periplaneta americana, or American cockroaches, was the study's population. Cockroaches had been
purchased from a controlled laboratory environment and had been bred intentionally at the University
of the Philippines Los Baños in Los Baños, Laguna. The cockroach samples used in this experiment
include both male and female specimens. Selection was conducted randomly without bias toward
gender, ensuring a representative and inclusive sample population. A similar approach was applied to
the criteria of sizes in choosing cockroach samples; no specific size of the insect was required as long as
they are the same species and are deemed adults.

The cockroaches were divided into positive, negative, and crude garlic extract groups without
randomization. The insecticidal impact of garlic bulb extract was evaluated by spraying the extract on the
treatment groups. Water and crude garlic extract were used in testing the control group, while Cyfluthrin
was used to test the positive control group. Before being subjected to the treatment, only the healthy
cockroaches were chosen; they were evaluated individually based on their activeness and alertness,
ensuring that each cockroach was fully functional before contact with the treatment.

The garlic plants that were used came from a single distributor and were collected not more than a week
before the extraction, ensuring the plant's freshness and reducing the possible factors that may affect
the condition of the plant. The size of the bulbs was not limited to a specific size range as long as they
were from the same source and underwent the same conditions.

Instrumentation

The main objective of this study was to assess the ability of compounds from garlic to exhibit
insecticidal effects when extracted from the garlic bulb to use against P. americana by determining the
mortality rate of the crude garlic extract. The data were collected during the experiment and analyzed
through Analysis of Variance (ANOVA), and the rate of mortality of cockroaches exposed to the extracted
garlic bulbs were measured.

Periplaneta americana was kept in an arena made from plexiglass, a transparent plastic that's often used
as a lightweight, shatter-resistant alternative to glass, with a height of 40.64 cm, width of 40.64 cm and a
length of 40.64 cm, each section has a same size of 40.64 cm both length and its width. A mesh fabric
was placed on top before the lid that kept them from escaping, and a lid made from plexiglass as well.
Hand spray bottles were used to precisely apply two (2) ml of the crude garlic (A. sativum) extract per
cockroach. Fifteen (15) cockroaches were placed in each container, and exactly 30 mL of the treatment
was applied.

According to Toryali et al. (2018), in their qualitative examination of the main phytochemicals and
bioactive components of Allium sativum in its aqueous solution have shown the presence of volatile oil,
glycosides, organosulfur, and saponins. Allicin, an important component that gives garlic its pungent
smell, which contributes to its insect repellent abilities, is predominantly connected to the elements
mentioned.

Materials and Methods:

Phytochemical screening of Allium sativum

The phytochemical composition of A. sativum was determined through a total of 18 ml of garlic crude
extract that was used for all three tests, namely, the Froth test for the presence of saponins, Sodium
Nitroprusside test, and Sulphur Test / Lead Acetate Test for the presence of organosulfur. These tests
were performed to identify the chemical constituents. Garlic mainly contains the sulphur components,
so two sulphur tests were performed and one for the saponin content.

A. Saponins: Froth Tests

2 ml of distilled water was added to the garlic extract in a test tube and shaken vigorously for 30
seconds. Allowed it to stand in a vertical position and was observed for 10 minutes. The presence of
saponins was confirmed by a soapy appearance in the mixture. Honeycomb froth greater than 3 cm
above the surface.

B. Sodium Nitroprusside Test

added 3 to 4 drops of sodium nitroprusside to 2 mL of the extract that had been taken out of the test
tube. After adding 2 to 3 drops of 40% sodium hydroxide, the medium became alkaline. The presence of
sulfur in the specific extract was revealed by the emergence of a deep violet color. To make sure there
were no oversights, these two tests were run three times each.

C. Sulphur Test / Lead Acetate Test

In a test tube, 2mL of extract were combined with 3 to 4 drops of lead acetate and 2 to 3 drops of 40%
sodium hydroxide. The mixture was then stirred until the precipitate was gone. 2 minutes were spent
boiling the test tube and then chilled. The presence of sulfur in the produced garlic extract was verified
by the formation of a brownish black precipitate.

To quantitatively assess the insecticidal effects of garlic on P. americana, plexiglass was treated with
positive (Cyfluthrin), negative (Water), and crude garlic extract solution. A precise volume (30 mL) of
each treatment group was evenly spread on the inner surface of the respective plexiglass section,
introducing it to the cockroaches. Groups of 15 cockroaches were placed in each container for 2 hours
and 25 minutes.

Data Gathering Procedure

This experiment followed certain procedures when they gathered the data. Experimentation was the
main tool that was used to gather data. It was conducted between two variables, namely: Garlic bulb
extract as the independent variable, and the insecticidal effect against American cockroaches as the
dependent variable.

Preparation of crude garlic bulb extract

Garlic plants were collected from Pantabangan, Nueva Ecija, from a garlic distributor in January 2025.
The bulbs were identified by the Bureau of Plant Industry (DA-BPI) at Malate, Manila.

The extraction of garlic bulbs was done at the laboratory of the Department of Pharmacy, Manila
Adventist College. The preparation of garlic bulb extract followed the method outlined by Azizah et al.
(2020) with modifications, where the extract was produced using a 1:1 ratio of garlic to water. For
testing, 100 mL of extract were needed. In a glass container, 1,000 grams of peeled garlic bulbs were
crushed, steeped in 1,000 mL of water for a whole day (24 hrs), and then stored at room temperature.
The garlic was soaked, then pressed with a fresh cloth to remove the extract from the crushed bulbs. The
resulting mixture was then filtered through filter paper and placed in a designated spray bottle.

Application on the American cockroach

The sources of the samples of P. americana came from the University of the Philippines Los Baños
(UPLB), located at Los Baños, Laguna, wherein the experimental group consisted of a total of 135
cockroaches. Three (3) sections of plexiglass with mesh fabric with the treatment groups (I) levels,
namely: I1 = positive control, I2 = negative control, I3 = crude garlic extract, where 15 cockroaches were
placed for 2 hours and 25 minutes, and monitored every 45 minutes. The test was conducted three
times, ensuring the results were consistent.

During the observation period, notes were taken based on the time and number of deaths after being
subjected to the treatment, then computed and analyzed the mortality rate and efficacy of each
treatment. After the data collection was completed, it was then analyzed statistically using the ANOVA to
determine the differences between the results. A table was used to illustrate the difference among the
treatment groups that have the highest mortality rate to the lowest mortality rate.

Post mortality observation and confirmatory test

After the exposure period, the mortality rate was recorded by counting dead cockroaches. The
data collected was calculated using ANOVA analysis based on the mortality rates for the three groups.
After collapsing, each cockroach has undergone confirmatory testing to endure their death. The test
included touching the insect on the abdomen using a stick. If the insect is unable to correct itself within
2 minutes or fails to show any movements, it is considered dead and counted for the mortality rate. This
test is based on the method used by Saipollizan et al. (2021) in their experiment.

Statistical Treatment of Data

A statistical method is employed in the research study to interpret the gathered data. The statistical
treatment of data aims to validate the general objectives and the progress of the research study.

1. Data collection:
 Mortality Rate: Percentage of American cockroaches killed after exposure in the garlic bulb extract,
water, and Cyfluthrin. The mortality percentage will be calculated using the formula M= (a/b) × 100%,
where ‘M’ represents the percentage of American cockroach mortality. ‘a’ denotes the number of
deceased cockroaches per treatment, and ‘b’ signifies the total number of cockroaches in each
statement.

Time of Mortality: How long it takes for the cockroaches to die after being exposed to garlic bulb
extract, water, and Cyfluthrin.

2. Group of Data;

Data is grouped into three categories (3):

A. Garlic bulb extract data which represents plant-based treatment.

B. Water data as a standard to prove efficacy.

C. Cyfluthrin data represents commercial insecticides.

3. Statistical test and analysis

The data collected based on the number of deaths of cockroaches in each treatment will be
subjected to ANOVA (Analysis of Variance) to compare mortality rates between garlic bulb extract, water,
and Cyfluthrin.

4. The tool and software

The Statistical Package for the Social Sciences (SPSS) will be used to analyze advanced statistics, as
we will use ANOVA.

Scoring and Interpretation

The scoring and interpretation criteria will allow the assessment of the insecticidal effects of different
treatments. To interpret the results properly and evaluate the efficacy of the extract from the
comparison between the results that will be obtained on subjects who will receive different treatments.

Treatment Number of Mortality rate every Number of total cockroach Mean

tested 45 minutes mortality after 2 hrs and 25
cockroaches mins (3 repetitions of 45
mins)
 T1 T2 T3 1 2 3 n (%)

I1 = Synthetic 15
Insecticide
(Positive control)

I2 = Water 15
(Negative control)

I3 = Crude Garlic 15
Extract

Table 3.1: Number of Periplaneta americana Mortality at Treatment groups of Crude Allium sativum
Extract After 3 repetitive 45 minutes of Treatment.

Score Range Percentage Category

0-3 1-20% Very Ineffective

4-6 21-40% Ineffective

7-9 41-60% Effective enough

10-12 61-80% Effective

13-15 81-100% Very effective

Table 3.2: Scoring table to interpret Mortality rate based on the number of deaths.

Scoring and Interpretation were adopted from a study done by Newyearsi et al. (2021)

Ethical Considerations

Ethical considerations were taken into account during the experimental study on insect subjects, in order
to maintain scientific integrity and humane treatment during the experiment.

During the research procedure, the well-being of the cockroaches were prioritized to mitigate any
potential harm or distress not associated with the insecticidal therapy. This commitment corresponds
with newly instituted protocols for invertebrate research, which underscore the need to nurture all living
organisms. All results, whether positive or negative, were disclosed in an open-access style to facilitate
information sharing and uphold scientific integrity. This study provided optimal environmental conditions
to the cockroaches to minimize stress, following the ethical guidelines for acclimatization by the
University of the Philippines Los Baños. Any potential discomfort before the actual experimentation was
minimized. Taking all issues into account, the study adhered to ethical standards and provided valuable
insights into pest management utilizing garlic bulb extracts as an efficacious natural insecticide.
 Chapter 4

Results and Discussion

Phytochemical test

Allium sativum contains different kinds of phytochemical constituents. However, the most
prominent of these bioactive compounds in garlic are alkaloids, tannins, flavonoids, phenolic
compounds, and organosulfur such as allicin, alliin, diallyl sulfide, diallyl disulfide, diallyl trisulfide,
ajoene and S-allyl-cysteine among many others (Shang et al., 2019, as cited in El-Saber Batiha et al.,
2020). The constituents we tested are the saponins and organosulfur of the extract. While the rest such
as its alkaloids, tannins, flavonoids, and phenolic compounds were not tested.

The garlic extract contains saponins such as furostanol and spirostanol saponins (Wang, et al., 2023).
Saponins' antifeeding, growth-regulating, and direct toxicity mechanisms work together to provide their
insecticidal effects. Saponins cause cytotoxicity at the molecular and cellular level by interfering with
membrane sterols. The reproductive organs, subcuticular layers, and the midgut are among the tissues
impacted by this method of action. In the cockroach's alimentary canal, saponins also interfere with
digestion and nutritional absorption (Shakeel et al., 2025).

Organosulfur compounds have been shown in some tests to be extremely toxic. They interfere with vital
enzymatic processes outside of neuronal targets. They could compromise vital metabolic processes by
interfering with cyclooxygenase and HMG-CoA reductase, which would affect cellular survival and
homeostasis. In exposed cockroaches, it causes oxidative stress by producing reactive sulfur species that
can harm organelles and cellular membranes, resulting in necrosis and tissue damage. Symptoms
observed in treated individuals include paralysis and necrosis, mainly in the stomach, the oral cavity, and
legs (Chaong et al., 2022).

The crude garlic extract exhibited a visible positive result from the sulfur test, showing a black precipitate
in all three repetitions which signifies that the extract contains sulfur. The crude garlic extract also shows
positive results from the saponin froth test. This concludes the presence of both saponins and
organosulfurs in the crude garlic extract. The positive results of the phytochemical results are
documented and can be seen in Appendix. HPLC was not used, therefore the exact amount of the
constituents are not measured.

Insecticidal effects on Periplaneta americana

Mortality was recorded and tabulated using the scoring and interpretation adopted from a study
done by Newyearsi et al. (2021) with modifications to accommodate the required information regarding
this experiment.

Treatment Number of tested Mortality rates every 45 minutes Number of total

cockroaches each cockroach mortality
test after 2 hrs and 25 mins

T1 T2 T3 1 2 3
 I1 = Synthetic 15 12 3 0 11 3 1 13 1 1 15 15 15
Insecticide
(Positive control)

I2 = Water 15 0 0 0 0 0 0 0 0 0 0 0 0
(Negative control)

I3 = Crude Garlic 15 0 5 5 3 5 6 2 1 6 10 14 9
Extract

Table 1: Scoring and Interpretation of the number of Periplaneta americana mortality from different
treatments.

From the results seen in Table 1, the positive control or I1 showed an outstanding insecticidal
performance as a synthetic insecticide while the negative control or I2 exhibits no insecticidal effect. The
crude garlic extract which is the I3 showed promising results as an effective botanical alternative for
chemical insecticides. The category of crude garlic extract will be categorized based on the number of
cockroach mortality per time intervals. Cockroach mortality is recorded every 45 minutes and only after
the confirmatory test to make sure which of the cockroaches are deceased and which are not.

The observed total mortality rate of the American cockroach when exposed to the extract, negative
control, and positive control is summarized using a statistical tool.

Positive control Negative control Crude garlic extract

Period Sum I Sum I Sum I

After 45 minutes 15 0 10

Another 45 minutes 15 0 14

Another 45 minutes 15 0 9

Table 2: Summarized results from each treatment.

The data provided in this paper give a positive indicator about the insecticidal properties of crude
garlic extract. However, the highest percentage of American cockroaches was obtained with the positive
control using synthetic insecticide. Comparative analyses will be used to determine the significant
differences between the three treatment groups to address the second research question of this study.

Using the Statistical tools, Friedman test and Wilcoxon Signed Ranks Test, results were analyzed to
compare the three treatments based on the number of mortality per time interval to determine the
significant differences between the efficacy of each treatment's insecticidal properties.

Groups Mean Rank Chi-square (df) Sig. Interpretation

 Positive control 2.33 11.88 (2) .003 Significant

Negative control 1.11

Crude garlic extract 2.56

Table 6: Summarized results from Friedman's test.

Based on the analysis using Friedman's test, it shows that there is a significant difference between
the negative control group, positive control group, and the experimental group using crude garlic extract.
This suggests that both control groups and the experimental group using crude garlic extract are
comparable. Therefore, the hypothesis is rejected or it is not accepted.

Groups Groups Mean Z value Sig. Interpretation

Rank

Negative control - Positive control Negative 4.50 - .011 Significant

ranks 2.536c

Positive ranks .00

Crude garlic extract - Negative Negative .00 - .011 Significant

control ranks 2.536c

Positive ranks 4.50

Crude garlic extract - Positive control Negative 7.00 -.423b .673 Not Significant
ranks

Positive ranks 3.00

Table 7: Summarized results of ranks based on the Wilcoxon Signed Ranks Test.

The Wilcoxon Signed Ranks Test shows a significant difference in cockroach mortality between the
negative control group (water) and the positive control group (Cyfluthrin). On the other hand, there is a
significant difference in cockroach mortality between the negative control group and the experimental
group given crude garlic extract. Additionally, there is no significant difference in the mortality of
cockroach positive control group and experimental group using crude garlic extract. This absence of
significant difference indicates that the crude garlic extract has insecticidal activity comparable to that of
a commercially available synthetic insecticide. Given these findings, crude garlic extract could be
regarded as a suitable botanical alternative for cockroach control. Its efficiency, combined with its
natural nature, distinguishes it as a safer and more environmentally sustainable alternative to synthetic
chemical pesticides, which are frequently associated with toxicity, environmental persistence, and the
development of pest resistance. As a result, the use of crude garlic extract has the potential to be an
environmentally friendly technique for integrated pest management programs targeting cockroaches.
 Hypothesis is rejected or it is not accepted for the negative control group and positive control as
well as the negative control group and experimental group.

Hypothesis is accepted for the positive control group and experimental group using crude garlic
extract.

Limitations of the Study

This study was performed to examine the insecticidal action of garlic crude extract on American
cockroaches (Periplaneta americana) and the results obtained were positive, but some limitations need
to be addressed. First, the study was conducted entirely in a controlled environment that does not
provide the complexity found in the field. The variation in temperature and humidity and what other
species or refuge may be present will all collectively impact the efficacy of garlic extract in the field. Also,
the work used only adult American cockroaches either male or female, which restricts the extent of
conclusions made to other cockroach species or household pests.

Another significant limitation is the potential difference in chemical composition of garlic based
on variations in the cultivar, cultivation conditions, and extraction processes. Such variations may
influence the consistency and replicability in the insecticidal activities of the extract. Furthermore, this
experiment considered the short-term or acute consequences of the garlic extract, while taking into
consideration any potential long-term consequences, like changes in behavior, reproductive impact, or
resistance that would manifest after some time, was not carried out.

The comparison of the efficacy of garlic extract with available insecticides limits the opportunity
to putatively relate to the potential efficacy of a garlic extract as a pest management tool. What was
applied to the insects as part of this study was a crude extract, and not a formulated product which
could have advantages for shelf life, odor, method of application, and user acceptability. Stability testing
of the garlic extract after at least weeks after the extraction could have also been examined. Finally, the
only phytochemical assays conducted were for organosulfur and saponins. Also, HPLC was not
performed, and again, some of the specific amounts for certain components of crude extract, were not
quantified. Overall, these omissions serve as a strong reminder that further research to look at the
greater applicability and pragmatic use of garlic insecticides are warranted.
 Chapter 5

Summary of Findings, Conclusions, and Recommendations

Summary of Findings

The results of observing the effectiveness of crude garlic extract as a botanical insecticidal effect
in Periplaneta americana showed that the crude garlic extract could be killed. The experiment presented
in this paper showed that garlic extract could have been highly effective as a natural insecticide against
cockroaches. When garlic cloves are crushed and formulated into sprayed or barrier applications, the
strong sulfur-based aroma repels cockroaches and disrupts their usual foraging and movement patterns.
Based on the laboratory tested, this further demonstrates how garlic sprays can elicit contact toxicity
leading to death when prolonged contact arises.

Garlic (Allium sativum), a member of the Liliaceae family, which contains chemical elements like
organosulfur and saponins capable of killing the cockroach (Periplaneta americana). It is very rich in
antioxidant phytochemicals, which include organosulfur compounds and saponins which are the
constituents used to prove the insecticidal properties of garlic in this study. Crude extract from garlic
reveals a significant potency as a botanical insecticide against species of cockroaches, which is a benefit
being exerted due to the contribution of abundant amounts of bioactive organosulfur compounds. These
compounds formed mainly from the degradation of allicin and sulfur-containing molecules after
maceration.

In this experimental study, it showed that within minutes after being in contact with crude garlic
extract, some cockroaches experienced slower movements compared to the negative group. The results
of this study showed that crude garlic was able to kill the cockroach within 45 minutes. Based on the
observations made during the contact toxicity testing, the crude garlic extract exhibited a positive
potential as a botanical insecticide. This crude extract could have been used as an alternative insecticide
to prevent or eradicate the population of American cockroaches (Periplaneta americana).

Conclusion

This study showed that garlic (Allium sativum) bulb crude extract exhibits insecticidal effectiveness
and displayed a potential as an botanical insecticidal against American cockroaches (Periplaneta
americana). The crude extract, Allium sativum, exhibits a significant insecticidal effect against
Periplaneta americana compared to negative control (water) and demonstrates a comparable effect with
the positive control. The statistical analysis revealed that there is no significant difference observed in
the mortality rate per time between positive control and crude extract. However, there was a significant
difference in the observed mortality rate per time interval between the positive and negative control and
the negative control and crude extract.

Recommendations

The presented study's results have demonstrated that crude garlic (Allium sativum) extracts have
commendable insecticidal activity against Periplaneta americana, thus providing hope for the potential
use of this in an effective eco-friendly manner against synthetic insecticides. Following this up, the
recommendation to future researchers is to conduct behavioral studies on cockroaches exposed to
garlic-based treatments. These ought to include careful observations regarding avoidance behavior,
locomotor patterns, and time to mortality that could yield more information about the action
mechanism of garlic compounds beyond mortality. Observed behavioral responses would also throw
light on whether garlic extract behaves as a repellent, a contact toxicant, or both, hence helping
formulate a more focused and efficacious pest control strategy. For a detailed basis for the formulation
of a pest control strategy in greater depth, any behavioral study in this domain should also describe
active moiety studies.

Also, for future researchers, there are opportunities for different concentrations, exposures, and
combinations of other botanical extracts to achieve and improve overall efficacy. Furthermore, garlic-
based botanical insecticides are a safer alternative to chemical insecticides that are environmental
detractors, are often toxic to non-targeted species, and create potential risks for humans. In comparison
to synthetic chemicals that contribute to resistance in pests and create bioaccumulation, botanical
insecticides such as crude garlic extract pose an environmentally reduced hazard and have a faster
degradation rate with lower toxicity properties. Therefore, they can contribute to sustainability for long-
term use. Future studies should also include the cost-benefit analysis and environmental impact for
using botanical formulations in agricultural settings and mostly in the household which the researchers
focus on in this study. The continuation of such research will help researchers and households may
contribute to the green pest control solutions which are effective while addressing public health and
environmental safety.