ANNONA SQUAMOSA LEAVES EXTRACT: A POTENTIAL DOG

GEL SOAP INGREDIENT FOR ERADICATION OF TICKS

A Research Paper
Presented to:
MS. RHONA BALDAGO
Of the Department of English
Senior High School
Lala National High School
Maranding, Lala, Lanao del Norte

In Partial Fulfillment
Of the Requirements for the Subject
Practical Research I
S.Y. 2019-2020

By:

BAUTISTA, TITO JR. V.

MELEGRITO, KEN SHANE
CUESTA, FLORESSAMIE JANE
LUMOSAD, ASHLEY JANE

August – 2019
 CHAPTER I
THE PROBLEM AND ITS SCOPE
Introduction
The Philippines, a country of more than 7,000 islands, is blessed with rare, exotic
flowering plants (Papaleng, 2016). The Philippine medicinal plant entrees now number
about 1100 (Stuart, 2019). The insecticidal and acaricidal properties of a number of plants
have been discovered long ago, and some of the plants can compete with synthetic
means of control (Hedin and Hollingworth, 2017). Especially remarkable are tropical
plants, from which hundreds of products of secondary metabolism with insecticidal
properties have been extracted (Hiiesaar et al., 2016). They are environmentally less
harmful than synthetic pesticides and acting in many insects in different ways. Custard
apple (Annona squamosa L.) extracts have shown promise for pest control against a
range of insect pests. Laboratory and field tests showed that extracts from custard apple
kernels were effective against crop pests like spotted stem borer, Chilo partellus (Swin.);
leafhopper, Nilaparvata lugens (Stal.), Spodoptera litura (Fab.), S. frugiperda,
Helicoverpa armigera (Hubner), hairy caterpillar, Spilosoma obliqua (Wk.); Brinjal spotted
leaf beetle, Henosepilachna vigintioctopunctata (Fabr.); cotton boll worm, Dysdercus
koenigii (Fab.); semi-looper Achaea janata Linn. and Aphids (Saito et al. 2016, Rao et al.
2018, Bhagawan et al. 2017, Ghatak and Bhusan 2016, Hiremath et al. 2015, Bhatnagar
and Sharma 2017, Mathew et al. 2019, Raman et al. 2016, Sonkamble et al. 2016,
Bhuiyan et al. 2016).
Ticks and fleas are common problems of both dogs and pet owners. Tick and flea
species are found worldwide. This study aims to produce soap that will help solve these
problems. Ticks are small but widespread arachnids that feed from blood of different
animals (including humans) throughout their life cycle and by doing so can transmit
pathogens (viruses or bacteria) that cause disease in humans. Although not all ticks are
infected, those that are cause a wide range of diseases worldwide – from bacterial
infections like borreliosis or tick spotted fever, to viral ones such as CCHF or even
parasitic infections like the poorly known babesiosis (Rodríguez, 2017).
In addition, ticks are becoming increasingly global. On one hand, they are more
active in more places and during longer periods of the year (beyond spring and summer,
their maximal periods of activity) due to climate change. On the other hand, migratory
birds, also affected by climate change, modify their flight routes and carry infected ticks
to places where they didn’t previously exist. Imported livestock can also contribute to the
spread of infected ticks. In addition, human encroachment of natural habitats for
agricultural activities, ecotourism and adventure travel increase the probability of contact
between humans and ticks. For international travelers, 14 tick-borne diseases have been
described (including Lyme disease, African tick fever, and tularemia) but the number will
likely increase with the emergence of new pathogens in new tourist areas (Rodríguez,
2017).
Recently, a phytochemical analysis in the laboratory of Department of Science and
Technology was conducted and it was proven that the leaves contain anthraquinones,
which is a derivative of anthracene that can be used as an insecticide. This test proves
that the Atis leaves extract contains an insecticidal property (Department of Science and
Technology, 2016). Herbal medicines include herbs, herbal materials, herbal
preparations, and finished herbal products that contain parts of plants or other plant
materials as active ingredients (World Health Organization, 2018). Using Atis extract as
an alternative treatment is an example of using herbal medicine. World Health
Organization states that the use of herbal medicines is safe and effective.
In Philippines, very few works have been done to explore the possibilities of
utilizing the local herbs and shrubs in veterinary practice. Therefore, the present research
has been designed to investigate the Annona Squamosa leaves extract as a potential
dog gel soap ingredient for eradication of ticks.
Statement of the Problem
This study aimed to investigate the Annona Squamosa leaves extract as a
potential dog gel soap ingredient for eradication of ticks. This sought to answer the
question:
1. Will the Annona Squamosa leaves extract be an effective way as an alternative
dog soap ingredient for eradication of ticks?
Scope and Limitations
This research sought to investigate the Annona Squamosa leaves extract as a

potential dog gel soap ingredient for eradication of ticks. This was limited in finding out
other uses of Annona Squamosa leaves extract other than its potential for eradication of

ticks.

Significance of the Study

In considering the effects of using social media among the Junior High in Lala
National High School, the researchers believed that the findings of this study will be
benefit the following persons:
Administrators. The result of this study will benefit the administrators of high
schools because this is concerned with the possible effect of Annona Squamosa leaves
extract as potential dog gel soap ingredient for eradication of ticks. They may use this
study as a reference to the teachers who facilitate their student’s research.
Teachers. From the findings, the teachers will be benefited from the study
because this study will reveal how Annona Squamosa leaves extract be an effective way
as eradicator of ticks. They can find more ways and to enhance students’ learning and be
able to use Annona Squamosa leaves extract as tick eradicator.
Students. Since this study investigate the Annona Squamosa leaves extract as a
potential dog gel soap ingredient for eradication of ticks, the students will also be aware
that Annona Squamosa can eradicate ticks instead of buying expensive tick eradicator
soaps. Therefore, if they want to use Annona Squamosa leaves extract to eradicate ticks,
it is good for them because they can save more money despite of using harmful
chemicals.
Parents. They may also benefit from this study since parents have a role to play
by supporting their children usually in their studies.
Researchers. This study may be used by researchers in coming up with
something that can contribute in the field of science especially in the effectiveness of
Annona Squamosa as tick eradicator. They may also pursue the researcher’s
recommended topics for further studies.
Definition of Terms
The researcher provides meaning of the terms that are used throughout the study
in order to have a better and clearer understanding of the study. These terms are defined
conceptually and operationally;
 Acaricidal. It refers to a pesticide that kills mites and ticks (Merriam-Webster
Dictionary, 2019). In this study, this refers to a pesticide that can kill ticks.
Arachnids. It refers to a kind of animal that has eight legs and a body formed of
two parts (Merriam-Webster Dictionary, 2019). In this study, this refers to ticks.
Encroachment. It refers to gradually move or go into an area that is beyond the
usual or desired limits (Merriam-Webster Dictionary, 2019). In this study, this refers to an
organism which it moves one place to another.
Insecticidal. It refers to a chemical substance for killing insects (Cambridge
Dictionary, 2019). In this study, this refers to a property of a specific plant that can kill
insects.
Kernel. It refers to a soft part inside the nut (Merriam-Webster Dictionary, 2019).
In this study, this refers to a part of the custard apple.
Parasitic. It refers to an animal or plat that lives in or on another animal or plant
and gets food or protection from it (Merriam-Webster Dictionary, 2019). In this study, this
refers to an organism which is attached with another organism.
Pathogens. It refers to something that can cause diseases (Merriam-Webster
Dictionary, 2019). In this study, this refers to a virus or bacteria that can be transmitted
form one organism to another.
CONCEPTUAL FRAMEWORK

Independent Variable Dependent Variable Output

Effective Annona
Eradication effect of
Squamosa leaves
Annona Squamosa
Extract of Annona Squamosa extract as potential
leaves extract as tick’s
leaves dog gel soap
eradicator
ingredient for
eradication of ticks
 CHAPTER II
REVIEW OF RELATED LITERATURE
This chapter presents the literature that will help in the analysis and will make us
know the effects of using social media to students. This literature are taken from a reliable
source in the internet.
The Philippine abounds with thousands of plant species of medicinal value. For
generations of rural to sub-urban fold, the use of herbal medicine has been main stay
treatment. This tradition takes its root several centuries back and has been preserved
through generations. With the continued escalation of health care, unabated rise of drug
prices, it is very clear that there is a need to find alternative ways to treat their conditions
(Mosqereda, 2015).
Instead of using synthetic drugs that can be bought in pharmacies, the Department
of Health (Department of Health, 2017) is convincing people to use traditional herbal
medicine. They attest that it is cheaper, accessible, and have little or no side effects.
In some Asian and African countries, 80% of the population depends on traditional
medicine for primary health care. In many developed countries, 70% to 80% of the
population has used some form of alternative or complementary medicine (e.g.
acupuncture).
Herbal treatments are the most popular form of traditional medicine, and are highly
lucrative in the international market place. Annual revenues in Western Europe reached
US$ Five billion in 2003-2004. In China sales of products totaled US$ 14 billion in 2005.
Herbal medicine revenue in Brazil was US$ 160 million in 2007(World Health
Organization, 2017).
The socioeconomic status of these cases shows no boundaries, but it stands to
reason that the poor, homeless, and children do account for the vast majority of the cases
because these groups traditionally find themselves with more opportunity for head-to-
head contact as a result of close living conditions, cohabitation, and simply the increased
population in a given geographical area. Age is a factor that plays a substantial role in the
occurrence of infestations, with children representing an abundantly large percentage of
the population with lice. When it comes to gender, studies vary as to whether or not there
is a statistical difference in incidence between boys and girls. Although not designed to
evaluate gender specifically, one study found a significant disparity, with girls being 7.4
times more likely to have lice (26.7% versus 3.6%) and 13 times more likely to be infested
with nits (23.8% versus 1.8%) than boys (n = 280) (Heukelbach, 2017).
According to Maglaya 2018, Environment plays a direct influence on the health of
people. Unsanitary environment is a major factor that causes problems like dengue,
diarrheal diseases, and Pediculosis. State of the country’s environment is the direct
result of the interaction of a number of factors such as industrialization, government
policies, poverty and an uncaring attitude towards the environment. Unfortunately, many
of us do not care for our environment. Community health nursing should therefore include
the environment in its health programs.
Potential for transmission is highest when people are in crowded, institutionalized
conditions or in war or prison camps, where sanitation is bad and clothing cannot be
changed often. Physical contact and stray hairs, even under sanitary condition easily
transmit them. (Schmidt & Roberts, 2018)
Annona squamosa Linn or what we call Atis here in the Philippines, belonging to
family Annonaceaeis commonly found in India and cultivated in Thailand and originates
from the West Indies and South America. It is mainly grown in gardens for its fruits and
ornamental value. It is known as custard apple, sugar apple, sweet après in English, and
sharifa in Hindi & sitaphalam in Telugu in India & corossolier & cailleux, pommier cannelle
in French (Barve, 2017).
Atis was introduced by the Spaniards at an early date. The plant is a small tree
with only a height of three to five meters. The leaves when young are hairy, oblong in
shape and eight to fifteen centimeters in length, with a petiole 1 to 1.5 centimeters long.
The flowers are pendulous, hairy, three-angled and greenish-white or yellowish. The fruit
is large, somewhat heart-shaped and six to nine centimeters in length. When the fruit is
already ripe it turns into yellow green (Burkill, 2019).
The Atis tree grows just about anywhere a seed fall. And it grows fast so that you
can get to harvest fruits in a little over a year from seedling stage. A tree bears fruits about
three times a year. They are in season from March to September, with June being the
peak (Department of Agriculture, 2019).
 There are many health benefits of Atis. The fruit contains calories, amino acids,
vitamins and minerals. Each 100 grams of the edible portion of Atis has about 50 mg of
ascorbic acid, 60 mg of lysine and 23-55 mg of phosphorus. The seed can be pounded
into paste that is applied to the head to get rid of lice, an annoying hair irritant (Philippine
Council for Health Research and Development, 2017).
Annona Squamosa L. or Atis leaf is a plant belonging to the family Annonaceaeis.
It is popularly cultivated in all parts of Thailand, especially in the northeast, as sweet fruit.
The seed of this plant is well known for killing head lice in many countries.
The human head louse (Pediculus humanus capitis) is a small insect causing a
public health problem, especially in poor sanitary conditions. In Thailand, research has
shown the anti-head lice activity of A. squamosa reported that the extract of custard apple
seeds in coconut oil at ratio of 1:2 could kill 98% of head lice within 2 hrs. while leaf extract
shows less potency (Chavasiri, 2016).
Atis is native to Tropical America and widely grown in arid regions of India. Its root
leaves and fruits are medicinally important. The leaves contain saponin and are
suppurative and insecticidal. They are used for treatment of ringworm (Taenia versicolor)
,destroying lice, proctoptitis in children and skin diseases, and as a poultice to produce
suppuration. It is also used as an anthelmintic, pediculicide, scabicide, and anti-
inflammatory. Person shaving hysterical on fainting fits are made to inhale crushed leaves
or their juice. Crushed leaves are applied to the nose area in cases of fainting spells.
Poultice of fresh leaves is useful in dyspepsia (Kumar, 2017).
In Indonesia, leaves of Atis are used as pediculicide. While in the Philippines the
leaves and bark of Atis are used as astringent. The ripe fruit is recommended in cases of
anemia. The seeds provoke vomiting. The powder of the seed, mixed with alcohol,
applied externally, is good for dandruff. The poultice from the fresh leaves is
recommended for dyspepsia. Crushed leaves can be applied to the nose to relieve
fainting spells. The unripe fruit and seeds have vermicidal and insecticidal properties. The
decoction of the root is a drastic purgative. Seed is used in the Philippines as a
pediculicide by crushing, mixing with coconut, and applying topically. In Thailand seeds
and leaves are used as a pediculicide. Crush 10-20 seeds (or 7-8 leaves, if seeds are not
available), remove seed coat, mix with animal or vegetable oil (1:2), strain and press to
obtain most of the oil. Apply the oil to the hair, cover with a thin cloth and wash thoroughly
after 1-2 hrs. (Best reported combination is ground seed mixed with coconut oil (1:2); it
kills 98% of lice in 2 hours; it is used externally to relieve scabies, ringworm and Tinea
versicoloe. A clinical trial showed that the seed and the leaf can treat hair lice with minor
side effects or irritation. The seed contains a caustic resin that is a toxic irritant principle
and is used to get rid of hair lice (Ecoport, 2016).
The leaves, fruit and the seeds contain acrid principle that possess vermicidal and
insecticidal properties. The crushed seeds is mixed with water as a paste and applied to
the hair scalp to destroy head lice. The seeds are also used as an abortifacient if applied
to the utero in pregnant women. The leaves with salt make a good cataplasm to induce
suppuration. The fresh crush leaves of atis can be applied to the nostrils to help relieve
fainting (Sanyal and Ghose, 2019).
Atis or sugar-apple (Annona Squamosa) leaves extract is the main variable in this
study. It contains substances that can repel insects or more specifically it can irritate and
remove lice. Recently, a phytochemical analysis in the laboratory of DOST was conducted
and it was proven that the leaves contain anthraquinones, which is a derivative of
anthracene that can be used as an insecticide. This test proves that the atis leaves extract
contains an insecticidal property. Such processes like preparation, filtration, squeezing,
and pounding was undertaken for the possibility of making a lice remover out of Atis
(Annona squamosa) leaves extract. As the researchers used the finished product to some
infected subjects, it was concluded, and that it was effective. Many lice were found in the
towel used to dry the hair of the infected subjects. The result of the study shows that lice
were removed on the infected subjects. Therefore, the researchers concluded that atis
leaves extract is useful and can be used as a lice remover to threat those who are
infected. Aside from the fact, it is cheaper compared to commercial products (Montoya,
2018).
According to Heong 2017, the insects should be homogenous, exclude all
variations such as age, sex, condition. Some insects such as beetle can be reared without
difficulty in large numbers; others such as human lice may be troublesome to produce in
quantity. With precise experimental conditions and insects difficult to rear batches as
small as 5 to 10 may be used. In allocating, it is best to apportion them in such a way that
the insects are randomized. Otherwise, the insects chosen first may all occur in the same
batch, and if their susceptibility is slightly abnormal; it will bias the mortality.
Parasite is specie that feeds on the body of another without killing it and generally
attacks only a single host. The eggs are usually laid upon the host. Among the insects,
there are two general groups of parasites; those that attack vertebrates and those that
attack insects. The former include (Anoplura) the bird lice (Mallophaga) some of the louse
flies (hippoboscidae) and the fleas (Siphonaftera) (Gullan, 2016).
Lice are parasitic insects that infest mammals. Infestation with lice is called
Pediculosis. Hundreds of varieties of lice infest humans. Three common kinds are
Pediculus Capitis (head lice) Pediculus Corporis (body lice) Pediculus Puris (crab lice).
Pediculus Capitis is found on the scalp and tends to stay hidden in the hair; the nurse can
suspect their presence in the clothing if (a) the person habitually scratches (b) there are
scratches on the skin and (c) there are hemorrhagic spot on the skin where the lice have
suck blood (Kozier and Erb’s, 2018).
Lice require blood from the host and feed several times a day. The victim is often
unaware of these silent passengers until itching occurs, which is a result of sensitization
to louse saliva, develops several weeks later. Scratching can result in secondary bacterial
infections. The head louse has legs especially adapted to grasp scalp hair. During a
lifespan of a little over a month, the female louse produces several eggs (nits) a day. The
eggs are attached to hair shafts close to the scalp. The very young stages of louse are
also called nits. Empty egg cases are whitish and more visible. They do not necessarily
indicate the presence of live lice. As the hair grows (at the rate of about 1cm a month),
the attached nit moves away from the scalp. A point of interest is that the incidence of
Pediculosis among blacks in the United States, lice have become adapted to the
cylindrical hair shafts found in whites. In Africa, lice have adapted to the not cylindrical
hair shafts of blacks (Tortora, 2018).
The adult louse is 2 to 3 mm long and usually light gray, although color may differ.
The female lives up to 3 to 4 weeks. After mating, an adult female louse lays one to six
eggs a day for up to one month until death. The eggs incubated by body heat, hatch in
10 to 14 days. Once the eggs hatch, nymphs leave the shell casing, breed for about 9 to
12 days, and develop into an adult lice and mate, and then females lay eggs. If not treated,
this cycle may repeat itself every 3 weeks. While living on the head, the louse feeds by
injecting small amounts of saliva and taking tiny amounts of blood from the scalp every
few hours (Fursule, 2019).
The term or span of life is limited for each species. Females usually live longer
than males and unmated females live longer than mated females. Food is an important
factor in determining the length of lice. Insects that have well developed mouth will not
live long if they do not receive food (Fundaments of Entomology, 2016).
Lice cannot survive for more than 48 hours away from a human scalp. Nits (lice
eggs) can survive for up to 10 days but need a blood meal from a human one-hour after
hatching. For that reason, human head lice cannot survive for long periods of time on
animals, but animals can be the carrier of the lice. For instance, if two children are playing
with a dog, the dog can 'transport' the louse from one child to another (Insight
Pharmaceuticals, 2016).
The nurse can suspect their presence in the clothing if (a) the person habitually
scratches (b) there are scratches on the skin and (c) there are hemorrhagic spot on the
skin where the lice have suck blood (Kozier and Erb’s, 2018).
If someone in your household or other close contact has lice or scabies, it is
possible for you to get them too. To prevent this from happening, the person that has lice
or scabies needs to be treated as soon as possible. It is also important to treat the whole
household. All clothing and bed linens that the person wore or came in contact with in the
two days before treatment should be washed in HOT water and dried in high heat. The
person’s combs and brushes should be soaked in rubbing alcohol or a disinfectant for
one hour. Finally, floors and furniture should be vacuumed (Organization of Teratology
Information Specialist, 2018).
A preparation containing 20% custard apple (Annona squamosa) seed extract was
shown to be highly effective against head lice infestations in a small controlled trial. More
than 90% of lice were dead three hours after application of the product to infested
schoolchildren, as compared to 60% dead lice in the control group, treated with 25%
benzyl benzoate emulsion. It has been used in traditional Thai medicine against
pediculosis since long ago (Spear, 2017).
 A filter paper diffusion bioassay method was carried out in order to determine the
licicidal activity of extracts. Petroleum ether extract of each plant tested individually,
showed high levels of mortality on adult lice, both plant extract showed significant
decrease in the mean time required to kill lice with 1% and 10% concentrations whereas
increase in the meantime was observed with 0.1% concentration when compared to 1%
lindane. Annona squamosa L. extract showed more potent activity than Azadirachta
indica A. extract at all concentrations 0.1, 1 and 10%. These results demonstrate the
possibility of using Azadirachta indica A. and Annona squamosa L. products for
controlling head lice by tribals in this area, which are difficult to control because of their
resistance to the currently used anti-louse agents. The assay proved simple, effective
and gave reproducible results (Investigation of licidal activity of some plants from Satpuda
hills; Kosalge, 2019).
A lotion that you will apply to the hair and scalp. It is approved to treat people ages
6 years and older. It works by paralyzing and killing the lice and eggs. When used as
directed, it is safe but it can irritate the skin a bit as it works. Some people get dry hair or
their skin can burn or sting (U.S Food and Drug Administration, 2019).
Pyrethrins are derived from daisy like flowers of pyrethrum, Chrysanthemum
cinerariaefolium. These are now obtained chiefly from East Africa, though they came
originally from Dalmatia and for many years prior to 1940 from Japan. The pyrethrins are
extracted from flowers and dissolved in oil such as deodorized kerosene for control of
household pest such as flies and cockroaches. The pyrethrins are available in mixtures
with DDT, upon insects. Insects frequently recover from the effects of pyrethrin poisoning,
so it is advisable to combine pyrethrins with some such as DDT to assure good kill. DDT
or dichloro-dipheny-trichloro-ethane is a stable, white, crystalline chemical which may be
prepared in form of a dust or oil. It is a definitely a poison and typical symptoms of tremors
may result from consumption of rather large quantities (Applied Entomology, 2015).
Treatment for pediculosis relies on conventional household cleaning practices and
the use of some neurotoxic pediculicide, which have been found to produce resistance in
some strains of head lice. These household practices may not be effective in decreasing
the Prevalence of pediculosis due to the problem of reinfection. Also, they are too
cumbersome to perform. Pediculicide chemicals, such as Permethrin and Malathion, were
also found to Induce resistance and allergic reactions among children (Philippine Science
Letters, 2016).
The number of new commercially available natural products for head lice has
expanded over the last decade to a much greater extent than products containing defined
chemical insecticides. However, the evidence on the efficacy of these new products
based on published results of in vitro and clinical trials is markedly deficient. Evidence on
safety is also deficient. All over, the counter natural products should be supported by in
vitro data and well-designed comparative therapeutic trials using head lice derived from
the populations for whom the product is intended. Since the prevalence and degree of
insecticide resistance of head lice to pyrethrin, permethrin, and malathion is expected to
increase, alternative topical therapies for pediculosis are needed. It is possible that, in the
long run, plant extracts, or their constituent compounds, will replace chemical insecticides
on the market (Canyon, 2017).
 CHAPTER III
RESEARCH METHODOLOGY
This chapter presents the materials needed, collection of Annona Squamosa
leaves, preparation of the extract, application of the extract and statistical analysis.
Materials
Annona Squamosa leaves
Jars
Covered jars
Butter cheesecloth
Open container
Small bottles
Eye dropper cap
Collection of Annona Squamosa leaves
Five hundred (500) grams of Annona Squamosa leaves will be collected in
Darumawang Ilaya, Lala, Lanao del Norte near in the house of the researcher. Only green
and fresh leaves will be collected. Additionally, these leaves are clean and dry to prevent
the other organic substance to be extracted in extraction process. If the leaves is wet, let
it air dry first before doing the extraction process.
Preparation of the extract
Wash your hands, soak the leaves to allow moisture to stay. It is important to pick
leaves that are green and moist to allow for easy extraction of the extracts. Get a jar put
the leaves and pours enough alcohol until it covers it. Cover so the alcohol does not
evaporate and shake it up until couple of days or so. After three to seven days, strain the
alcohol from the solids and transfer it to a cover jar. Place the solids back to the original
jar, cover with fresh and put the lid on. After three to seven days strain out the alcohol as
before from the second extraction. Combine it with the alcohol from the first extraction
and filter it all through butter cheesecloth to remove any remaining solids. Place the
alcohol in an open container and allow it to evaporate for a few days. After the alcohol
evaporates, you’ll be left with the oils. Put the oils into small bottles with eye dropper cap.
Application of the extract
And test the gel soap for its effectiveness in killing the ticks and fleas of a dog by spreading
the gel soap to its whole body, scrubs it for 2-3 minutes and drain/cleanse it. And after
that we will measure the amount of ticks and fleas died by using new product we’ve made
and record the data in the data notebook.
Statistical Analysis
All data gathered were processed and analyzed by Mean Analysis.
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