UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 1

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Allium sativum as a Natural Cockroach Repellent (Periplaneta

americana)

A Research
Presented to the
Senior High School
University of Santo Tomas

In Partial Fulfillment
of the Requirements of the Course
Practical Research 3

by

Alexis Nicole Lyica G. Paule

Sabrina V. Periabras
Francesca V. Quirino
Gabriel L. Quiling
Rex Christian DR. Plaza

12STEM-14

Month Year
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ABSTRACT

According to the World Health Organization, cockroaches are the

unhygienic scavengers in human settlements. Moreover, these also carry

Salmonella species, which can cause gastroenteritis and typhoid, and

Poliomyelitis, which causes polio. Other bacteria from cockroaches can also cause

dysentery that may lead to severe diarrhea with blood. Periplaneta americana is

the most common among these cockroaches in the Philippines, which is 35–

40mm in length and shiny reddish to chocolate brown color. Chemical

insecticides are known to practically repel these insects, but these repellents may

have adverse effects in humans because of their toxicity. Nowadays, organic

repellents have been taken as a healthy control measure. The most common

organic repellent uses Nepeta cataria or Catnip, which is not usual in the

Philippines (widely used in Europe and North America). The researchers seek to

develop a natural repellent which is made from garlic (A. sativum) that is widely

grown in the Philippines. This study evaluates the characteristics of A. sativum

and assesses its effectivity as a cockroach repellent (specifically in P. americana).

The researchers observed the cockroaches exposed to the A. sativum solution for

one day exposure categorized in different time periods, 1st hour, 6th hour & 24th

hour, within two trials in under the same conditions. The observations were
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time-lapse recorded, categorized in different time periods (1st, 6th, 24th hour).

The raw data were analyzed by statistically calculating the mean, percentage

repellency from the formula of Sakuman and Fukami, and excess proportion

index (EPI). The analysis shows that as the number of hours increase, the

repellency percentage also increases. Percentage repellency for trial 1 of the

cockroaches treated with A. sativum for trial 1 is 88.89%. Trial 2 yielded to

86.67% percentage repellency. In conclusion, garlic is an effective repellent in the

resistance of American cockroaches (P. americana) through prolonged exposure

to the garlic solution. Further improvements in its repellency and residual

application may be ascertained with consideration to the modification of its

solution. This study can be furtherly used for future references and

improvisations regarding natural repellents.

Keywords: repellency, EPI, Allium sativum, Periplaneta americana

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TABLE OF CONTENTS

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1.0 Introduction
1.1 Background of the Study 4-5
1.2 Statement of the Problem 5-6
1.3 Objectives 6
1.4 Significance of the Study 7
1.5 Scope and Limitations 8
1.6 Acknowledgements

2.0 Review of Related Literature

2.1 Allium sativum
2.2 Garlic’s Antibacterial properties and Repellent effects

2.3 Benefits of garlic as a repellent

2.4 Periplaneta americana
2.5 Cockroaches in Filipino Households

2.6 Cockroaches as Carriers of Bacteria

2.7 What Repels Cockroach?

3.0 Methodology
3.1 Periplaneta americana samples
3.1.1 Collection of samples
3.1.2 Identification of Periplaneta americana

3.2 Allium sativum solution

3.3 Efficacy
3.3.1 General procedure

3.3.2 Repellency test

4.0 Results and Discussion

5.0 Conclusion and Recommendations

5.1 Conclusion
5.2 Recommendations

References 40
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Appendix I: Timetable for Research 61
Appendix II: Budget Proposal 62

CHAPTER 1

INTRODUCTION

1.1 Background of the Study

According to Shahraki et al., in Southeast Asia, P. americana were found

to be the most dominant species. They are in constant search for food and are

carriers of bacteria and pathogens. It was observed that the Gram-negative

dominant species were Escherichia coli, Klebsiella species and Enterobacteria

species known to cause diseases and direct transmission of infection

(Chaichanawongsaroj et al., 2004).

Many methods have been used to repel and control cockroach problems

in the community. The most commonly used are insecticide sprays that can

effectively eliminate them but because of the chemical composition of the

insecticide spray such as toxicity, they are potentially hazardous to humans,

animals, and other organisms and the environment. Therefore, people who use

pesticides regularly or come in contact with these must understand the relative

toxicity, potential health effects and preventative measures. Due to the given risk

factors, people are looking for an alternative to control the cockroach problems

in their household. Therefore, turning to natural ingredients that would not pose

any risks to the health and at the same time be as effective. Garlic (A. sativum) is
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known as a powerful insect repellent that can be used to repel a variety of

crawling and flying insects. While synthetic repellents are being used extensively

for reduction of tick infestations on hosts, there is a growing awareness of the

risks associated with the rampant use of these substances. Consequently, there is

mounting interest in the use of alternative and environmentally friendly

arthropod pest control agents such as plant-based repellents. A. sativum is

attracting interest from researchers as a potential source of tick repellent,

especially following a report that the consumption of garlic protected soldiers

exposed to ticks from tick bites under field conditions. Also, there are many

anecdotes and ethnobotanical claims suggesting that garlic extracts repel pest of

arthropod origin (Nchu, et al., 2016).

1.2 Statement of the Problem

Cockroaches are one of the most common and allergenic of indoor pests.

Not only do they are associated with indoor dirt, but cockroaches can affect

breathing. They also carry a diverse pathogenic bacteria flora, different

protozoans, pathogenic worms, fungus, and viruses which can have a detrimental

effect on those prone to asthma and allergies (Do, et al., 2016).

Cockroaches can also contaminate food found in abundance near wet and

damp corners and areas of the household. Cockroach control must be measured

to disrupt cockroach life cycle, yet pesticides to repel cockroaches contain

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hazardous chemicals which may bring health risks and side effects through

inhalation, skin contact, and ingestion.

Number of insect repellents have been developed with the idea of

consumer benefits. In addition to already known advantageous application of

insect repellents, there is increasing concern about the potential toxicity in

environment leading to health caused by random use of these compounds.

Insect repellents may also trigger undesirable hazardous interactions with

biological systems with a potential to generate harmful effects (Roy, et al., 2017).

This research seeks to study the efficacy of natural A. sativum to repel

cockroaches in households that has a non-toxic mode of action and degrades rapidly

that has no adverse effects on humans.

1.3 Objectives

The study aims to determine the efficiency of A. sativum and assess its

effectivity as a cockroach repellent.

This study specifically aims to answer the following questions:

1. to test and determine if A. sativum has repellency effects through

performing repellency test on P. americana;

2. to know the effects after applying the A. sativum solution, and;

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3. to determine if A. sativum solution can help withstand the prevalence of

cockroaches in Filipino households.

1.4 Significance of the Study

This research aims to determine the efficacy of a natural product, A.

sativum, as a cockroach repellent. It is to maintain, develop, and use natural and

nontoxic resources to reduce detriments in health and in the environment.

Therefore, this study is mainly for the sustainability of the environment.

This research is for households to divest cockroaches, which are also pest

insects that are abundant in human dwellings, where some species can carry a

diverse pathogenic bacteria flora, different protozoans, pathogenic worms,

fungus and viruses (Tatfeng et al., 2005).

This research also aims to produce a natural repellent that will minimize

the harmful environmental damage caused by chemical substances− to protect

homeowners, users, and consumers from the chemicals present in synthetic

insecticides that lead to different adverse effects both to humans and their

environment.
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It is for the community for resourcefulness and to take a rise on

improvisation to garlic as a cockroach repellent. This research is also conducted

in order to build future references regarding this type of topic. It is also for

students who will attain the full completion of the research towards cockroaches

and repellents.

1.5 Scope and Limitations

The research took place in Sampaloc, Manila, Philippines. This study is

only limited to the extract of Garlic (A. sativum) applied to help in eliminating the

American Cockroach (P. americana). This research is to contribute to ways in

eliminating cockroach (P. americana) pests. Other species of cockroach, insects,

and pests are not included in this study and only Garlic (A. sativum) extract

serves as an independent variable.

The estimated time of the experiment from the searching of cockroaches in the

neighborhood to the testing of the efficacy of garlic on cockroaches may range from a minimum

time of 6 hours to 48 hours that will depend on the chance of finding a proper test subject and

the time it would take for the A. sativum solution to take effect.
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CHAPTER 2

REVIEW OF RELATED LITERATURE

2.1 Allium sativum

Garlic, A. sativum L. or ‘bawang’ is a perennial herb of the Amaryllidaceae

family. The plant produces bulb which is surrounded by sheaths and composed of

thin-shelled bulblets, cloves, or set, all of which can form a new plant. The

bulblet in fresh or in processed form has been used as food, condiment, and for

medicinal purposes. Aside from being an indispensable recado (ingredient), garlic

is also used to cure athlete’s foot and hypertension. It is used in preparation of

drugs, insecticide and explosives. The crop is widely cultivated in the Ilocos,

Southern Tagalog and central Luzon regions. All varieties grown in the Philippines

are of native origin. These are Batangas white, Ilocos white and Batanes white

(Mamaril, 2010).

According to Majewski (2014), Garlic (A. sativum) is one of the most

researched and best-selling herbal products on the market. For centuries it was

used as a traditional remedy for most health-related disorders. Garlic’s properties

result from a combination of variety biologically active substances which all

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together are responsible for its curative effect. The compounds contained in

garlic synergistically influence each other so that they can have different effects.

A lot of variation among garlic products can be sold for medicinal purposes.

Clinically, garlic has been evaluated for number of purposes, including treatment

of hypertension, hypercholesterole-mia, diabetes, rheumatoid arthritis, cold or

the prevention of atherosclerosis and the development of tumors.

Garlic has been used both as spice and medicine for thousands of years.

With the present craze on use of natural products, health maintenance or as

therapeutic agents, garlic, in its various preparations, has been popularly used for

high blood pressure, for high cholesterol, and for heart disease. It is also used to

prevent certain types of cancer such as stomach and colon cancers (Sia, 2008).

Medicinal plants such as thyme, onion, blackseed, lemon balm and nettle

are intensively used in traditional medicine, today. Garlic is among the most

important of these plants. Garlic is reported to be a wonderful medicinal plant

owing to its preventive characteristics in cardiovascular diseases, regulating

blood pressure, lowering blood sugar, and cholesterol levels, effective against

bacterial, viral, fungal and parasitic infections, enhancing the immune system and

having antitumoral and antioxidant features. It contains sulfur compounds

(allicin, alliin and agoene), amino acids such as cysteine, glutamine, isoleucine

and methionine which help to protect cells from the harms of free radicals, and

vitamins C, E, and A which help to protect us from oxidation agents and free
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radicals, and other vitamins such as niacin, B1 and B2 and betacarotene (Ayaz et

al, 2007).

2.2 Garlic’s Antibacterial properties and Repellent effects

Since the discovery and exploitation of antibiotic agents in the

20th century, the targeted selective toxicity of such agents has ensured their

widespread and largely effective use to combat infection; however, it has

paradoxically resulted in the emergence and dissemination of multi-drug

resistant pathogens. Antimicrobial resistance in garlic in both medicine and

agriculture is now recognized by the World Health Organization (WHO), along

with other various national authorities, as a major emerging problem of public

health importance (Woods-Panzaru et al., 2009).

According to Benkeblia (2004), onion and garlic are among the first of the

cultivated crops in the world due to their long storage time and portability. They

could be dried and preserved for several months. At the present time, the Allium

family has over 500 members, each differing in appearance, color and taste, but

close in biochemical, phytochemical, and neutraceutical content. Alliums were

revered to possess antibacterial and antifungal activities and contain the

powerful sulfur and other numerous phenolic compounds which arouse great.

Onions and garlic are composed mainly of water (85–90 g/100 g and 60–70

g/100 g fresh weight, respectively) and the most significant components,

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medicinally, are the organo sulfur-containing compounds. However, garlic

contains nearly three times as much sulfur-containing compound as onions (11–

35 mg/100 g fresh weight). The mature, intact Alliums contain mainly cysteine

sulfoxides, and when tissues are chopped, the enzyme allinase is released,

converting the cysteine sulfoxides into the thiosulfinates. These compounds are

reactive, volatile, odor producing and lachrymatory. During the last 50 last years,

protection of food from spoilers and pathogens aroused great interest and was

achieved by various physical and chemical methods. Various garlic preparations

have been shown to exhibit a wide spectrum of antibacterial activity against

Gram-negative and Gram-positive bacteria including species of Escherichia,

Salmonella, Staphylococcus, Streptococcus, Klebsiella, Proteus, Bacillus, and

Clostridium. Even acid-fast bacteria such as Mycobacterium tuberculosis are

sensitive to garlic. Among these numerous and abundant naturally occurring

compounds, essential oil extracts have been considered as natural preservatives

or food additives and can be used as additional methods of controlling pathogens

(Benkeblia, 2004).

A wide range of microorganisms including bacteria, fungi, protozoa and

viruses have been shown to be sensitive to crushed garlic preparations. Analysis

of steam distillations of crushed garlic cloves performed over a century ago

showed a variety of allyl sulfides. However, it was not until 1944 that Cavallito

and his colleagues isolated and identified the component responsible for the
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remarkable antibacterial activity of crushed garlic cloves. The compound turned

out to be an oxygenated sulfur compound which they termed allicin, from the

Latin name of the garlic plant, A.

sativum. Pure allicin is a volatile molecule that is poorly miscible in

aqueous solutions and which has the typical odor of freshly crushed garlic. Final

proof of the chemical structure of allicin came in 1947, when it was shown that

allicin could be synthesized by mild oxidation of diallyl disulfide. Moreover, in the

clove, alliinase is found closely associated with a lectin (natural defense on

plants). The site of linkage of the carbohydrate moieties of alliinase has been

identified at Asp 146. Significant homology has been reported between the garlic

and onion alliinases although alliin was not detected in the latter species (Ankrl

and Mirelman, 1999). Lectins show its effect right from sensory receptors of

mouth parts by disrupting the membrane integrity and food detection ability.

Subsequently, enter the gut lumen and interact with midgut glycosylated

proteins like alkaline phosphatase (ALP), aminopeptidase-N (APN), cadherin-like

proteins, polycalins, sucrase, symbionin and others. These proteins play critical

role in life cycle of insect directly or indirectly (Upadhyay et al., 2012). Garlic

cloves are odor-free until crushed. Cross-section studies have indicated that the

substrate alliin and the enzyme alliinase are in different compartments. This

unique organization suggests that it is designed as a potential defense

mechanism against microbial pathogens of the soil. Invasion of the cloves by

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fungi and other soil pathogens begins by destroying the membrane which

encloses the compartments that contain the enzyme and the substrate. This

causes the interaction between alliin and alliinase that rapidly produces allicin

and which in turn inactivates the invader (Ankrl and Mirelman, 1999).

Akballah and Suleiman (2014) said that the bactericidal effect of fresh

garlic juice was tested by Louis Pasteur onto growing bacterial colonies. It was

shown that fresh garlic juice inhibited the growth of most gram-positive and

negative bacteria. The action was comparable in vitro with penicillin,

streptomycin, chloramphenicol, tetracycline and erythromycin. Previous studies

further demonstrated that ethanol and aqueous extract exhibited growth

inhibition of most gram positive and negative as well as multi- drug resistant

bacteria. The chemistry of the Allium species has been dominated by many sulfur

containing compounds that give them a characteristic flavor and exhibit potent

anti-fungal properties. Among the most studied are allicin, alliinase and allin (S-

allyl cysteine sulfoxide), the precursor of allicin which upon crushing of garlic

bulb, it is hydrolyzed by the enzyme allinase to its active form allicin. It is

responsible for most of the biological property of garlic. One gram of allicin (S-

methyl-l-cysteine sulfoxide) has been equated to 15 IU of penicillin in its

antibiotic activity. However, a variety of non-sulfur compounds, work

synergistically to provide various health benefits. Freshly pressed juice of garlic

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has a strong antifungal effect on the major pathogenic molds, yeast and

dermatophytes.

Garlic (A. sativum), natural fungicidal properties that work effectively to

control pests, makes an excellent economical, non-toxic biological pesticide for

use in agriculture. The natural pesticide (garlic) has a strong aroma that can

provide an olfactory camouflage against insects, masking their normal host-

finding or feeding cues. Aphids, ants, termites, white flies, beetles, borers,

caterpillars, slugs and army worms are some of the pests that can be suitably

controlled using garlic (Magwenya T. et al., 2016).

Lyme borreliosis is the most common vector-borne disease in Sweden,

and as many as 10000 individuals are thought to be affected each year. Studies

suggested that diethyltoluamide is the best repellent against insect vectors and

permethrin against ticks. However, insect repellents may have adverse effects on

humans and animals. Because military personnel are at particularly high risk for

tick bites and tick-borne diseases, we conducted a prospective, randomized,

double blind intervention trial of garlic (A. sativum) to prevent tick bites among

Swedish marines. Of 100 individuals in Swedish military service in 1998, 50

consumed 1200 mg/d A. sativum in capsule form and 50 consumed placebos for

8 weeks, followed by a washout period of 2 weeks, and then a crossover to

placebo or A. sativum consumption for another 10 weeks. All participants wore

the same type of uniforms, consumed approximately the same diet, participated
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in similar activities, and spent equal amounts of time in tick-endemic areas. Tick

bites were recorded in a diary after daily self-inspection of the skin. Written

informed consent was obtained from all participants. The study was approved by

the Research Ethics Committee of Lund University and the Swedish Medical

Products Agency. Data were analyzed by both intentions to treat (involving all

participants present at the start of the study) and per protocol (only the 80

individuals who completed the study) (Stjenberg and Berglund, 2000).

In the intention-to-treat analysis, 66 (66%) of 100 participants recorded

tick bites vs 55 (69%) of 80 participants in the per-protocol analysis. A total of

286 tick bites were recorded by the participants. On average, the participants

recorded 0.2 tick bites per week during military service, compared with 0.03 tick

bites during leave. There was significant reduction in tick bites when consuming

garlic compared with placebo in per protocol analysis (Wilcoxon test, P=.04). A

greater number of the participants were bitten by ticks during placebo

consumption (normal approximation of binomial assumption, relative risk by

intention to treat, 0.79 [95% confidence interval {CI}, 0.65-0.96]; relative risk per

protocol, 0.70 [95% CI, 0.54-0.90]) (Stjenberg and Berglund, 2000).

Considering the loss of storage products and adverse effects of chemical

pesticides, the use of plant extracts is one of the best methods for controlling

storage pests. Therefore, many plants essential oils and crude extracts have been
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evaluated for their insecticidal and repellence properties against field crop and

stored grain pests (Stjenberg and Berglund, 2000).

According to Debra & Mischeck (2014), herbs such as onion (Allium cepa)

and garlic (A. sativum) have been used in the intercropping systems because of

their ability to repel insects. Cole crops grown next to garlic and onion are less

prone to insect pest attacks. Onion and garlic plants produce excretions from

their roots as well as aromas from their leaves. These excretions and aromas

have beneficial effects on surrounding plants. They will discourage insects and

are therefore regarded as insect repellent plants. Natural pesticides are safer and

eco-friendly. In addition, they reduce cost of health management on farms

because they reduce inhalation of toxic fumes of synthetic chemicals. Garlic (A.

sativum) and onion (A. cepa) is widely grown and is easy to cultivate in gardens.

Garlic is reportedly effective against a wide range of disease-causing pathogens

and insect pests at different stages in their life cycles. Allicin is a compound

obtained from garlic. The compound is a colorless liquid which has a distinctively

pungent smell which reveals anti-bacterial and anti-fungal properties. Allicin in

garlic imparts defense mechanism against attacks by insect pests.

Plant extracts obtained from garlic (A. sativum) has a broad spectrum of

biological activity. Their anti-inflammatory, antibacterial and antifungal activities

are well known. Extracts from these plants possess also anti-insect properties.

Methanolic extracts of garlic have been reported to be larvicidal against larvae of

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several species of mosquitoes, antifeedant and insecticidal effects. Garlic extracts

have been reported to exhibit insecticidal activity against aphids M. persicae and

Brevicoryne brassicae (Dancewicz & Gabryś, 2008).

Upadhyay et al. (2012), said that the lectins or lectin-like compounds

(ASAL) in garlic may interfere with different aspects of the insect life cycle. Lectins

serve as plant defenses against insects, viruses, fungi, bacteria and mites. These

garlic compounds are toxic to many insects and can be a strong deterrent to

feeding and egg laying behavior. They bind to and damage sensory receptors,

midgut receptor proteins, intracellular proteins and other vital proteins within

the insects’ digestive tract.

2.3 Benefits of garlic as a repellent

Pesticides are widely used by developing countries such as the

Philippines for agricultural purposes. However, pesticide poisoning is still one

of the global health problems. It is estimated that about 5 million people die

every year as a result of intentional, accidental, and occupational exposures

worldwide (Lu, J. et al., 2010).

The repellent and anti-feeding effect of garlic oil was evaluated in

laboratory conditions against the bite of Phlebotomus papatasi females. The

effectiveness was evaluated by two different laboratory procedures: topical

application of garlic oil on five human volunteers, using the "standard cage test",
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and feeding sandflies on artificial membranes treated with the compound. Garlic

oil showed a significant protection by topical application on the skin of

volunteers, being the protection 97% and 40%, respectively at 1% and 0.005%

dilution. Garlic oil showed also an anti-feeding effect when tested on the artificial

membrane feeding apparatus; the anti-feeding effect was dose dependent, being

100% at 1%. The results of the present study confirm previous observations on

the repellent effect of garlic oil against haematophagous arthropods

(Vijayalakshmi, et al., 1999).

Garlic (A. sativum) has long been known to have uses in pest control for

its repellent effects. There is strong evidence to show it has great medicinal value

for its antibiotic, antibacterial, anticoagulant, and antiseptic properties (it was

called Russian penicillin in World Wars I and II) and it is often used as a cold, sore

throat, and flu remedy. Used as pesticide garlic has a non-toxic mode of action.

Garlic is not persistent in the environment since it degrades rapidly and has had

no adverse effects on humans. Formulated as a powder, distilled extract from

garlic cloves, or as an oil spray, garlic may be useful for pest control in some

situation. Organic gardeners have long been familiar with the repellent or toxic

effect of garlic oil on pests. Some studies also suggest that a garlic oil spray has

fungicidal properties. The repellent, insecticidal, nematicidal, fungicidal and

antibiotic properties of garlic has proved scientifically (Vijayalakshmi, et al.,

1999).
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Unlike synthetic pesticides which have only one active compound and

exhibit only one type of biological effect, single plant derived compounds may

have more than one biological effect. Detailed investigations have revealed that

several plants species have more than compounds and have diverse biological

effects. The chances of quick development of resistance to different chemicals in

one extract responsible for either one biological effect or different biological

effect one highly unlikely (Mathew et al., 2014).

2.4 Periplaneta americana

Cockroaches may become pests in homes, schools, restaurants, hospitals,

warehouses, offices and virtually in any structures that has food preparation or

storage areas. They contaminate food and eating utensils, destroy fabric and

paper products and impart stains and unpleasant odour to surface they contact

(Mathew et al., 2014).

Cockroaches are one of the oldest known winged insects and maintain

close contact with humans. Approximately thirty of the over four thousand

species of cockroaches found to date are harmful to humans. Generally,

cockroaches exist in environments with large amounts of toxic substances,

including pollutants, microbial toxins, insecticides and other xenobiotics. One of

the most common domestic species of cockroaches is the American cockroach (P.

americana), also known as Water bug or Palmetto bug (Zhang et al., 2016).
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The cockroach P. americana, is an obnoxious and notorious pest of the

world, with a strong ability to adapt to a variety of complex environments.

According to R. C. Perrott (2010), This specific specie of the cockroach is the

largest of the house-infesting roaches in households. This type of cockroaches

has been spreading throughout the world by commerce as P. americana

maintains close contact with humans and exists in environments with abundant

toxic substances (Zhang et al., 2016). American cockroaches prefer to live in

humid and tropical environments, like conditions created by humans. In line with

this is the increasing rate of their infestation in the Southeast Asia (Shahraki et

al., 2013).

The American cockroach is a large cockroach. The adult body length is

approximately 1-1/2 inches long (4cm). The antennae extending from the head

are equally as long as the body if they are intact (Perott, 2010). The female

produces an egg capsule (ootheca) that may protrude from the body for a few

hours up to four days and are then deposited near a food source. A total of 16

eggs per capsule are possible, but the average number is ten. Over the span of

about a year, an American cockroach can molt, or shed its skin, as many as 13

times before reaching maturity. Cockroach adults can then live for up to a year,

making the whole life cycle about two years under the best conditions. Because

they live in sewers, trash compactors and landfills, cockroaches are known to

carry bacteria and other diseases on their bodies (Alpert & Frye, 2014).
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As stated in the study of Perott (2010), when American cockroaches

aggregate, their presence is primarily an aesthetic nuisance. However, members

of this species are also known to carry infectious bacteria on their bodies and in

their gut. These bacteria may be transferred to food and other items that the

cockroaches contact. Several bacteria commonly associated with American

cockroaches are known to cause food poisoning, dysentery, and diarrhea in

humans.

2.5 Cockroaches in Filipino Households

In the study of Pai et al. (2005), P. americana and Blattella germanica

were collected from 40 households in Kaohsiung City and Kaohsiung County,

Taiwan. Cockroach infestation was found in 50% of the studied households and

226 cockroaches (123 P. americana and 103 B. germanica) collected by trapping.

P. americana was more often found in the kitchen (70.7%) whereas B. germanica

in the storage room (51.5%) and kitchen (36.9%). There was no significant

difference between the percentages of P. americana (99.9%) and B. germanica

(98.0%) carrying bacteria. A total of 25 species of bacteria was isolated from P.

americana and only 21 from B. germanica.

According to the study of Sia Su et al. (2016), there are currently 127 true

cockroach species from 5 families are recorded in the Philippines, 12 of which are

introduced while 63 are endemic to the country. Several new country records

were noted. All cockroaches collected in the streets of Metro Manila were
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taxonomically identified as P. americana as the distal segment of its cercus is

elongated, length more than twice its width. The insect color is reddish brown

and a pale brown to yellowish band around the edge of their pronotal shield. The

wings of the male extend beyond tip of abdomen and have caudal tergite that is

deeply notched, distal portion of plate thin projecting as hood over its

corresponding terminal sternite. This study has assessed the parasites infesting

on the external surface of cockroaches obtained in the streets of Metro Manila.

The occurrence of parasites in the cockroaches examined indicates the need to

raise people’s awareness that they need to safeguard themselves and their food

sources from being contaminated by cockroaches, as it may play an important

role in the mechanical transmission of parasite-related morbidities. Proper

personal hygiene and proper environmental sanitation must always be observed

to reduce the possible risks that exposure to the microorganisms harbored by

cockroaches may bring in our communities.

2.6 Cockroaches as Carriers of Bacteria

Cockroaches are insects recognized to be mechanical vectors of

disease-causing agents (Sia Su et al., 2016). It is stated that a great variety of

insects are carrier of pathogens. Cockroaches (Diploptera punctata) are

known to carry a diverse pathogenic bacteria flora, different protozoans,

pathogenic worms, fungus and viruses, but their role in the direct
 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 25
PAGE
transmission of infection has seldom been established and will be found in

abundance near leaking faucets or pipes, wet sponges, persistently damp

corners, areas where there is frequently standing water, or areas where

continual moisture is usually available, such as drains, kitchens, bathrooms

and maintenance rooms with sink traps. Water and steam pipes frequently

serve as migration routes from room-to- room. The filthy breeding habits,

feeding mechanisms and indiscriminate travel between filth and food make

some groups of synanthropic insects such as nonbiting flies and cockroaches

efficient vectors of human enteric protozoan parasites. Twenty-one species of

filth flies have been listed by regulatory agencies concerned with sanitation

and public health as causative agents of gastrointestinal diseases based on

synanthropy, endophily, communicative behaviour and strong attraction to

filth and human food. Outbreaks and cases of food- borne diarrheal diseases

in urban and rural areas are closely related to the seasonal increase in

abundance of filth flies, and enforced fly control is closely related to

reductions in the occurrence of such diseases (Tatfeng et al., 2005).

The World Health Organization has described cockroaches as

important pests because they spread filth and ruin food, fabrics and book-

bindings. They disgorge portions of their partially digested food at intervals

and drop feces. They also discharge a nauseous secretion both from their
 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 26
PAGE
mouths and from glands opening on the body which give a long-lasting,

offensive cockroach smell to areas or food visited by them.

Cockroaches move freely from building to building or from drains,

gardens, sewers and latrines to human habitations. Because they feed on human

feces as well as human food they can spread germs that cause disease. They are

proven or suspected carriers of the organisms causing: diarrhea, dysentery,

cholera, leprosy, plague, typhoid fever, or viral diseases such as poliomyelitis. In

addition, they carry the eggs of parasitic worms and may cause allergic reactions,

including dermatitis, itching, swelling of the eyelids and more serious respiratory

conditions (World Health Organization, 1997 ).

Cockroaches, insects commonly seen as household pests, are generally

regarded as a nuisance. However, the medical importance of cockroaches has

been largely overlooked though they have been shown to harbor a number of

pathogenic and non-pathogenic micro-organisms. Due to their omnivorous and

voracious feeding habits and their indiscriminate defecation and regurgitation

over human food, cockroaches are implicated in the spread of several infectious

agents that humans acquire by ingestion (Brown & Alhassan, 2015).

Various bacteria may simply be carried on the insect's cuticle or be

ingested and, some time later, regurgitated or excreted. Moreover, several

species of bacteria of public health significance have been isolated from, or have

passed through, cockroaches (P. americana) and their digestive tract, such as
 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 27
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Staphylococcus aureus, Streptococcus spp., Enterobacteriaceae, Pseudomonas

aeruginosa, and others. Numerous pathogenic bacteria, including Salmonella

spp., Shigella spp., Campylobacter spp., Pseudomonas aeruginosa, and Klebsiella

pneumoniae have been isolated from cockroaches and these insects greatly

contribute to food-borne disease outbreaks. As a result, insects like cockroaches

collected in hospitals and households have been found to harbor multidrug

resistant (MDR) bacteria and hospital cockroaches with drug-resistant Klebsiella

spp. have been suggested to play a role in the epidemiology of nosocomial

infections. In addition, a neonatal unit infested with cockroaches suffered an

outbreak of nosocomial disease due to extended-spectrum β-lactamase-

producing K. pneumoniae (Moges et al., 2016).

Cockroaches are likely to be encountered in favorable environmental

conditions with a ready source of food. The range of problems caused by the

presence of cockroaches vary between hospitals, food-handling establishments,

public institutions, and multi-family dwellings. In hospitals, it was found that

cockroaches can act as potential vectors in the epidemiology of nosocomial

infections, especially the transmission of drug-resistant bacteria. Escherichia coli,

Pseudomonas aeruginosa, Klebsiella spp and several other potential pathogens

have been isolated from cockroaches collected from hospitals. Cockroaches have

been associated with an outbreak of dysentery. Salmonella typhi, Shigella

dysenteriae and toxigenic strains of Escherichia coli can be retained in the gut of
 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 28
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cockroaches for up to several days. Transmission could occur by cockroach

regurgitation or fecal pellet deposition into human foodstuffs. Thus, these insects

also play a role in transmitting food-borne diseases. Their presence reduces the

perception of human well-being and sanitary standards (Chaichanawongsaroj et

al., 2004).

Cockroaches (Order: Blattodea) are important vectors of pathogens due

in part to their unsanitary lifestyle. Cockroach cuticle can harbor several

Enterobacteriaceae species including Salmonella spp., Klebsiella spp., and

Escherichia spp. A few medically important pathogens that can be vectored by

the American cockroach, P. americana (Linnæus) (Blattodea: Blattidae), include:

Campylobacter spp., E. coli, Salmonella spp., Shigella spp., Staphylococcus spp.,

Streptococcus spp., and Toxoplasma gondii (Barcay 2004). Cockroaches are also

able to transmit pathogens such as anthrax, cholera, diphtheria, pneumonia,

tetanus, and tuberculosis. Many of which could be used as bioterrorism agents

targeting animal or human populations (Pechal et al., 2007).

2.7 What Repels Cockroach?

Bug repellents and pesticides or chemical substances that deter,

incapacitate, ill, or otherwise discourage any pest from coming in contact the

covered object. Of bug repellents in pesticides the target pest includes insects,

parasites, plant pathogens, weeds, mollusks, birds, fish, nematodes and microbes
 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 29
PAGE
that destroy property, because nuisance, spread disease, or are disease paths

(Walte, 2017).

According to Coats et al., (2003), a repellent composition compromising

an amount of monoterpenoid or sesquiterpenoid in a combination with a carrier

is disclosed. In one embodiment, the monoterpenoid or sesquiterpenoid is from

a biorational source such as plant volatile. In a particular embodiment, the plant

volatile is a monoterpenoid, such as "nepetalactone" (or the individual

nepetalactone isomers) derived from catnip (Nepeta cataria). In another

embodiment, the plant volatile is any one or a combination of sesquiterpenoids

derived from the fruit of the orange tree (Maclura Pomifera) Such compositions

have repellency against anthropod, such as cockroaches, mosquitoes, mites,

ticks, spiders, and so forth method of repelling comprises applying an effective

amount of a composition comprising the compound together with a suitable

carrier in or near a target area, including applying the composition directly onto

humans, animals (e.g. pets, livestock), and so forth.

As stated by Behan & Bich (2003), certain compounds have long been

known to possess insect deterrent properties, some of this information coming

from what might be termed as folk knowledge. These materials include widely

known substances such as citronella, tolu, and Peru Balsams. Materials known

for their deterrent properties include those having camphoraceous odors such as

Camphor itself, Cypress oils, Galbanum etc.

 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 30
PAGE
A method for attracting cockroaches to an object or area, involving

treating said object or area with a cockroach attracting composition containing a

cockroach attractant effective amount of at least one compound selected from

the group consisting of methyl, ethyl, propyl, or butyl esters of C 3-7 saturated or

unsaturated carboxylic acids and mixtures thereof, and optionally a carrier

material or carrier. A method for repelling cockroaches from an object or area,

involving treating said object or area with a cockroach repelling composition

containing a cockroach repelling effective amount of at least one compound

selected from methyl, ethyl, propyl, or butyl esters of C 3-7 saturated or

unsaturated carboxylic acids or mixtures thereof, and optionally a carrier

material or carrier (Vandermeer et al., 2012).

The fumigant toxicity of various essential oils and monoterpenoid

components were tested against the adult German cockroach. Responses were

varied according to plant material, concentration, and exposure time. High

insecticidal activity against adult B. germanica specimens was achieved with allyl

isothiocyanate (component of horseradish) and the essential oil of A. sativum.

The findings show that alllyl isothiocyanate and A. Sativum had high insecticidal

activity, and that they have potential as fumigants for use against the German

cockroach (Tunaz et al., 2008).

 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 31
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CHAPTER 3

METHODOLOGY

3.1 Periplaneta americana samples

3.1.1 Collection of samples

15 individual species of P. americana were obtained from Dexae’s pet

shop in Sta. Ana, Manila, Metro Manila manually using trapping method. The

trapping element was constructed from an empty jar, petroleum jelly and starch:

the cockroaches are attracted to the jar by bread and raisins placed at the

bottom, and a thin layer of petroleum jelly inside upper rim which will prevent

the insects from escaping. P. americana was used as they are common in the

households in the Philippines. The samples were kept in a box maintained at 25-

28˚C standard room temperature. The box was also surrounded by a thin layer of

petroleum jelly in the lid to prevent the cockroaches from escaping. The P.

americana samples were fed ad libitum with starches such as bread, biscuits and

raisins. Sheets of paper were placed on the substrate to prevent the stress from

insects.
 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 32
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3.1.2 Identification of Periplaneta americana

The adults measure to an average length of around 4cm and about 7mm

tall; reddish brown with a yellowish margin on the body region behind the head;

and have immature stages that resemble adults except they are wingless

(Ejimandu et al., 2015).

3.2 Allium sativum solution

The chemistry of the Allium species has been dominated by many sulfur

containing compounds that give them a characteristic flavor and exhibit potent

anti-fungal properties. Among the most studied are allicin, alliinase and allin (S-

allyl cysteine sulfoxide), the precursor of allicin which upon crushing of garlic

bulb. It is hydrolyzed by the enzyme allinase to its active form allicin. It is

responsible for most of the biological property of garlic (Benkeblia 2004).

The antimicrobial activities of garlic and other plant alliums are primarily

based on allicin, a thiosulphinate present in crushed garlic bulbs (Wallock-

Richards et al., 2015).

To make the repellent solution, 85 grams of crushed and minced A.

sativum was mixed thoroughly with 950mL of distilled water, 50mL of olive oil.

Distilled water was preferred to avoid impurities that could take effect to the

repellent. Olive oil was used to help stick the solution to the target area. The

measurements were based from the study of Vijayalakshmi, Subhashini, and Koul
 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 33
PAGE
of Plants in Pest Control: Garlic and onion. The soap was eliminated from the

materials used to lessen unnatural ingredients in the solution. The solution was

left for 15 minutes to let the mixture rest briefly. During this resting period, the

liquid absorbed the garlic. Hence, the solution was still shaken before using to

avoid oil from floating. A 3x3 cheesecloth was soaked in the garlic solution and

sat for another 15 minutes.

3.3 Efficacy

3.3.1 General procedure

Efficacy was studied through repellency test. An apparatus was

improvised as a box which is a mimic of an ordinary household with four corners.

It has dimensions of; 7.5 inches height, 15 inches width, and 20 inches length.

Cockroaches was introduced in to the box. They were observed for the first 15

minutes Then the soaked cheese cloth was placed on the center of the box.
 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 34
PAGE

The apparatus was observed for 24 hours. The observations were

recorded for the first hour, sixth hour, and twenty-fourth hour. The number of

cockroaches under NT (number of insects draw near the proximity chemical-

treated cloth) and NC (number of insects away from the chemical-treated cloth)

were listed per hour of observation. This timetable and experimentation we used

was acquired from the Study of Repellent Activity of Different Plant Powders

against Cockroaches (P. americana) of Mathew, Reshma, and Rejitha.

3.3.2 Repellency test

The number of insects trapped in the chemical-treated cloth and the

number of insects away in the control test chamber of both results from each

specie was recorded. From the obtained data repellency was calculated using the

formula of Sakuman and Fukami.

 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 35
PAGE

Where:

EPI : excess proportion index

NT : number of insects draw near the proximity of the chemical-treated

cloth

NC : number of insects away from the chemical-treated cloth

PC : Percentage repellency (percentage of animals trapped in control test

chamber)

CHAPTER 4
 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 36
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RESULTS AND DISCUSSION

A. sativum was assayed for its repellant property against cockroach, P.

americana. Observations were made by exposing insects to the A. sativum

solution for one day. The different exposures were categorized in different time

periods: 1st hour, 6th hour, and 24th hour. The results were analyzed and were

based on the repellant activity exhibited by the cockroaches. The hesitation of

the insects to move at the center of the apparatus, where the A. sativum solution

is placed, is considered its repellent effect. Two trials were done, and both were

under same conditions. Two trials were set to be conducted to evaluate that the

results didn’t happen by chance. The number of cockroaches which approached

the area of the solution was observed and data were calculated statistically by

excess proportion index and percentage of repellency of Sakuman and Fukami

(1985).

The number of insects leaving the treated cloth compared to the control

were the determining factor of the repellence activity of the A. sativum. From

the gathered data, the A. sativum showed its repellent property. Repellent

activity increases the potential value of A. sativum as a natural repellent to P.

americana. Excessive Proportion Index (EPI) values give an idea about the

repellency or attractancy of the chemical substance against an animal tested. A.

sativum showed both attractant and repellent activity.

 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 37
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Mathew et al. (2014) said that the Excessive Proportion Index ;(EPI) was

classified as: if values <1 it indicates repellency; if 1 it is neutral; and if >1 the

material used is attractant in nature. The study of Mathew et al. (2014) also

explained that some studies show that sometimes, the reason for these types of

insects’ behavior might be due to the presence of an olfactory stimulus. The

presence of volatile compounds having strong odor could block the tracheal

respiration of the insect therefore, leading to their death.

Table 1: Number of NT and NC (where NT: number of insects draw near the

proximity of the chemical-treated cloth and NC: number of insects away from the

chemical-treated cloth)

15 P. americana TIME DURATIONS

(treated with A.
1st HOUR 6th HOUR 24th HOUR
sativum)
NT NC NT NC NT NC
Trial 1 3 12 2 13 0 15
Trial 2 4 11 2 14 0 15

The observations were time-lapse recorded, which were categorized in

different time periods (1st, 6th, and 24th hour). The frequency of the cockroaches

which approached the solution in the center was shown. Table 1 shows the

number of insects which draw near the area of the chemical-treated cloth (NT)

and the number of insects (NC) which were repelled from it. The two trials were

recorded and the first hour has the highest frequency on both trials. The 24 th
 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 38
PAGE
hour has the highest number of insects which were repelled from the chemically-

treated cloth. As the number of hours increased, the NC also increased.

Sulfur-containing compounds, stated by Benkeblia, give A. sativum a

characteristic flavor and exhibit potent anti-fungal properties. Among the most

studied are allicin, alliinase and allin (S-allyl cysteine sulfoxide), the precursor of

allicin is the crushing of the Garlic bulb, it is hydrolyzed by the enzyme allinase to

its active form allicin. These support the repellent activity of A. sativum with the

present findings of the study.

Table 2: Percentage repellency of P. americana treated with A. sativum for 1st, 6th,

and 24th hour.

A. sativum TIME DURATIONS

treatment st
1 HOUR 6th HOUR 24th HOUR
PC EPI PC EPI PC EPI
TRIAL 1 80% -0.6 87% -0.7 100% -1
TRIAL 2 73% -0.5 87% -0.7 100% -1

The percentage repellency was calculated through the formula of

Sakuman and Fukami (1985). Percentage repellency is the percentage of

cockroaches which are repelled from the proximity of A. sativum treated cloth. In

trial 1, the percentage repellency increases from 80% in the first hour, 87% in the
 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 39
PAGE
sixth hour, to 100% in the 24 th hour. In trial 2, the percentage repellency varies

from 73% in the first hour, 87% in the sixth hour, and 100% in the 24 th hour.

Percentage repellencies for the 1st, 6th, and 24th hour were accumulated

per trial. The total percentage repellency for trial 1 is 89%. Trial 2 yielded to 87%

percentage repellency.

The negative value of EPI indicates that the number of insects trapped in

the chemical-treated test chamber is less than the number of insects trapped in

the control test chamber. This implies that EPI and Percentage repellency have an

inverse relationship as shown in table 2. In the experiment, as the number of

hours in exposure increases, EPI value gradually reduced from -0.6 to -1 in trial 1

and -0.5 to -1 in trial 2. Thereby, this gives the values of percentage repellency

from 80% to 100% in trial 1 and 73% to 100% in trial 2. As the time duration

differs (1st hour, 6th hour, and 24th hour), the percentage repellency and EPI also

varies. Due to their inverse relationship, as the time duration progresses, the

percentage repellency increases while the EPI decreases.

 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 40
PAGE

CHAPTER 5

CONCLUSION AND RECOMMENDATIONS

5.1 Conclusion

Through the prolonged exposure of P. americana (American cockroaches)

to the Garlic solution, A. sativum is proven effective in repelling P. americana.

The percentage repellency of the treated cockroaches increased as the time

duration (1st hour, 6th hour, and 24th hour) progressed. The repellency effects of
 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 41
PAGE
A. sativum as a natural cockroach repellent were determined through the

Percentage Repellency (PC) and Excess Proportion Index (EPI). The analysis

indicated the repellency effects of A. sativum. In conclusion, A. sativum can aid

withstand the prevalence of cockroaches in Filipino households as an effective

repellant.

5.2 Recommendations

This study determined the characteristics of A. sativum and assessed its

effectivity as a cockroach repellent (specifically in P. americana). Based on the

results and conclusion of the study, the researchers have recommendations for

improvements of future studies regarding natural repellents. The researchers

suggest to use a larger version of the apparatus in order to mimic an ordinary

household. Through this, observations on the behavior of cockroaches will be

improved. The future researchers must also utilize a more suitable equipment

and methods to observe the cockroaches. The behavior of the cockroaches at

any period is important to the findings and analysis of the research. Precise and

accurate measurements shall be observed in gathering data and recording

observations. Consequently, it is recommended to have a thorough background

information of the chemical properties and biological activities of the main

variables before starting and editing the paper works and experiment itself.
 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 42
PAGE

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Appendix I Timetable for Research

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(Example)

Research Objectives 2010 2011

Jun Jul Aug Sept Oct Nov Dec Jan Feb Mar Apr May

1. Writing of Proposal
2. Collection of Algal
Materials
3. Fractionation of
Polysaccharides
4. Collection of Blood and
Isolation of PBMCs
5. Cell Viability Assay
6. ELISA Assay for Cytokines
7. MRNA extraction
8. Chemopreventive assays
9. Data Collation/Analysis
10. Final Defense
 UNIVERSITY OF SANTO TOMAS SENIOR HIGH SCHOOL 54
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Appendix II Budgetary Requirements

(Example)
SOURCES OF EXPENSES Quantity Amount
(PhP)
Collection …….

Isolation of ….

Tissue Culture Reagents

Total