CHAPTER I

THE PROBLEM AND ITS BACKGROUND

INTRODUCTION

This remains a worldwide public health problem with an estimated nineteen

percent prevalence of head lice among school children and two percent prevalence of

pubic lice in adult populations. This information is based on the research conducted in

May 2022 by Yi-Tian Fu, Chaoqun Yao, Yuan-Ping Deng, Hany M. Elsheikha, Renfu

Shao, Xing-Quan Zhu & Guo-Hua Liu.

Human blood-sucking lice has two families, Pediculidae and Pthiridae, with the

corresponding genera Pediculus and Pthirus, respectively. Pediculus humanus capitis

represents the body louse or clothes louse while Pthirus pubis refers to the pubic louse

or crab louse. Head lice spend their entire life on the host, body lice live mainly on the

folds of the host’s clothing and bedding. Head lice do not favor socioeconomic classes,

but body lice are often observed on homeless and are associated with poverty,

overcrowding and poor hygiene. Crab lice prefer thick coarse hair, such as pubic hair,

but can also infest other body locations. Transmission of head and body lice occurs via

close contact, from head-to-toe such as exchanging hats, sharing pillowcases, and

sharing clothes. Meanwhile pubic lice are transmitted from person to person via direct

skin-to-skin contact.

. A Research study year 2011 found that neem seed extract successfully killed

head lice larvae after 5 minutes of treatment and adult head lice after 10 minutes of

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treatment, this may be due to azadirachtin content. Azadirachtin has been identified as

neem’s principal active compound. It acts on insects by repelling them, disrupting their

growth, metamorphosis, and reproduction. Neem-based solutions do not usually kill

insects directly, but they can alter their behavior in significant ways to reduce their

reproductive potential. It has been shown to affect egg production and hatching rates. In

lice, azadirachtin can cause them a reproductive disability where they can no longer

infest in a person’s hair and will eventually die. Azadirachtin is mainly found from the

Neem oil which can be extracted from the plant Neem seeds.

Our research mainly focuses on treating head lice infestation which is the

Pediculus humanus capitis, or also known as the body and head louse. We, the

researchers, aim to make a solution to aid people who suffer from this health problem

by using organic plants that are easy to get from our natural resources. In doing so, this

will be more cost-effective and pose less risk to the health of the consumers. The

researchers invented a plan to create a bar of shampoo that is mainly composed by the

neem oil that can be found inside the neem seed. We, the researchers, wanted to try

our best to create a lice bar shampoo to test its effectiveness out of neem seed oil for

treating head lice infestation. Our main goal as a team is to aim for success of this study

from the start to the very end.

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 CONCEPTUAL FRAMEWORK

INDEPENDENT VARIABLE DEPENDENT VARIABLE

 Azadirachta Indica  Bar shampoo for

(Neem) seeds and treating lice infestation.
leaves.

 Respondents

Table 1. Conceptual framework of the study showing the relationship of the identified

variables.

The table above shows the independent and dependent variables for this

research study. The independent variable is the plant Azadirachta Indica seeds (and

leaves) which is the main material to be used to produce bar shampoo, and the

respondents to check the effectivity of it. The dependent variable is the product that is to

be created out of the Neem plant. There is yet no tests included in the independent

variables since this research is not yet conducted as laboratory research.

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 RESEARCH PARADIGM

INPUT PROCESS
 The use of Azadirachta  Extraction of neem oil.
Indica (Neem) as a
treatment for lice  Preparation of other
infestation. materials needed for
making the bar shampoo.
 Preliminary investigation

 Experimental observation
 Data gathering

OUTPUT
 Azadirachta Indica (Neem) bar shampoo

Table 2. Research paradigm shows the concepts of the researchers from input,

and process to the output of the researchers. It provides the researcher’s overview of

the process, the testing, and the output. The use of the main ingredient, for preliminary

investigation and data gathering as input of the study. The process includes the

collection of Azadirachta Indica (Neem) extracts and preparation for the process of

making shampoo and experimental observation. The Azadirachta Indica (Neem) as a

Treatment for Lice Infestation is the output of the study.

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STATEMENT OF THE PROBLEM

This research is conducted to know the probability of Azadirachta Indica (Neem)

as bar shampoo for treating lice infestation.

Specifically, this research will answer the following questions:

1. How effective Azadirachta Indica (Neem) as bar shampoo for head lice?

2. What are the possible effects of Azadirachta Indica (Neem) in lice?

3. What are the other benefits of using Azadirachta Indica (Neem) bar shampoo?

ASSUMPTIONS

 The Respondents will rate the product truthfully.

 This study will be a success.

 The product will be a complete success.

 The effectiveness of Instruments used.

HYPOTHESIS

 Null Hypothesis:

 Alternative Hypothesis:

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SIGNIFICANCE OF THE STUDY

The significance of this study is to provide people with more knowledge regarding

the potential of Azadirachta Indica (Neem) as a Treatment for Lice Infestation.

Especially for the benefit of those who are infected of lice infestation, to aid them by

creating a product without the use of any harmful chemicals to avoid causing harm to

the consumers. Additionally, it will be beneficial for them as it is more cost-effective than

other options currently in the market. It will also benefit the future Researchers as they

could use this study as their reference to improve what we have started in this study in

creating a product out of a neem tree to treat lice infestations in human hair.

SCOPE AND DELIMITATIONS

This research covers the Problem, Research locale and Time frame. This

research aims to create a solution for lice infestation without the use of harmful

chemical that can cause anyone’s health harmed. Hence, we will use Azadirachta

Indica (Neem) as the main material to create a bar shampoo that kills lice.

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Problem/Scope: The scope of this study is limited to the making of Azadirachta Indica

(Neem) as bar shampoo for treating lice infestation, with other materials to be used

throughout the process.

Research locale: This research will be conducted at Holy Trinity University.

Time Frame: This research study started in February to April 2023.

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DEFINITION OF TERMS

Antiarthritic - relieves or prevents arthritic symptoms, such as joint pain and joint

stiffness.

Antibacterial - kills bacteria or stops them from growing and causing disease.

Antifungal - used for treating fungal infections, which most commonly affect skin, hair,

and nails.

Anti-gastric ulcer - it is for the prevention of gastric ulcer.

Anti-hypoglycemic - an agent to lower glucose levels in the blood.

Anti-inflammatory - reduces inflammation such as redness, swelling, and pain in the

body.

Antipyretic - used to prevent or reduce fever.

Antitumor - inhibiting the growth of tumor or tumors.

Azadirachta Indica – commonly known as neem, it is a tree native to the Indian

subcontinent, but can also be found around the Southeast Asia.

Azadirachtin – it is a chemical compound belonging to the limonoid group, is a

secondary metabolite present in neem seeds.

Head lice – also known as Pediculus humanus capitis, are tiny, wingless insects that

spend their entire lives on the human scalp and feeding exclusively on human blood.

Lice infestation – also known as pediculosis, it is a common condition caused by

parasitic insect Pediculus humanus capitis.

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Metamorphosis - it is a biological process by which an animal physically develops

including birth transformation or hatching.

Shampoo bar – a solid form of the traditional hair cleanser.

Therapeutics - treatment used to alleviate or prevent particular disease.

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 CHAPTER II

REVIEW OF RELATED LITERATURES

FOREIGN LITERATURE

The neem tree is primarily cultivated in the southern regions of Asia and Africa

where it has been seen used through many ages in medical folklore. We should note

that various parts of the neem tree, including the leaves, bark, fruit, flowers, oil, and

gum are associated with the aforementioned medical folklore in the treatment of certain

medical conditions such as cancer, hypertension, heart disease, and diabetes. The

potential effects that are seen when using these extracts can certainly be attributed to

both cellular and molecular mechanisms. These mechanisms include free radical

scavenging, detoxification, DNA repair, cell cycle alteration, programmed cell death

mitigation, autophagy, immune surveillance, anti-inflammatory, anti-angiogenic, and

anti-metastatic activities. They also provide the ability of modulation of various signaling

pathways. (Arumugam et al., 2014, Omóbòwálé et al., 2016, Patel et al., 2016).

The Azadirachta Indica (neem tree) was traditionally employed intensively as a

folklore remedy for a wide spectrum of diseases in India (Mulla and Su, 1999).

Azadirachta Indica has a wider arrary of uses than any other herb. The first recorded

use of neem is attributed to the ancient East Indian ‘Harrappa Culture’ which added the

plant to dozens of health and beauty aids 4500 years ago. The centuries old healing

system, Ayurvedic medicine, has utilized these timeless neem solutions as a mainstay

of Ayurvedic pharmacy. Its medicinal qualities are outlined in the earliest ‘Sanskrit’

writings that state use of various parts of Azadirachta Indica to treat bacterial, fungal,

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and viral infections which in turn boost the immune system. Also, its usefulness as a

natural non-toxic insecticide among other fascinating properties increases it

phenomenal applications (Wealth of India, 2000). Numerous scientific reports validate

the traditional uses of neem in both the maintenance of general health and skin care.

Practically every part of Azadirachta Indica (leaves, bark, fruit, flowers, oil, and gum)

have been reported to be associated with various remedial properties such as,

antimicrobial effects (Sai Ram et al., 2000), storage behavior (Sacande et al., 2000),

reduction of paracetamol-induced liver damage (Bhanwara et al., 2000), enhancer of

hepatic glutathione and glutathione-dependent enzymes (Arivazhagan et al., 2000), in

vitro antiviral activity (Badam et al., 1999), insecticidal activity (Et-shazly and et-

sharnoub, 2000), antibacterial agent (Das et al., 1999), etc. Chemopreventive potential

of Azadirachta Indica has also been evaluated recently on 7,12-dimethyl

benz(a)anthracene (DMBA)-induced hamster buccal pouch carcinogenesis (Balasenthil

et al., 1999) and against IBD virus in broilers (Sadekar et al., 1998).

FOREIGN STUDY

As for the study that aimed to evaluate in vitro-effectiveness of shampoo

formulation containing neem oil as an anti-head louse. An experimental method was

used to examine the formulation of shampoo, irritation test, testing the effectiveness of

shampoo against head lice (Pediculicidal activity), and testing the effectiveness of

shampoo against nits (Ovicidal activity).

The effectiveness test employed an in vitro-method where neem oil shampoo

was tested directly towards head lice and nits isolated from humans, then the mortality

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was observed. To recognize the safety of the shampoo, skin and eye irritation tests

were carried out by applying 0.5 ml on the skin and dripping 0.1 ml shampoo into New

Zealand white rabbit’s eyes. Data were analyzed using Kruskal Wallis and Mann

Whitney.

The study results showed that the primary irritation index for 5%, 10%, 15%, and

20% neem oil shampoo were 0; 0111; 0.222; and 0.222, respectively. All of the

shampoos were very mild irritation response category. Eye irritation tests showed that

only the 20% neem oil shampoo gave a mild irritation response with degree of corneal

opacity ≥ 1 and recovered after 7 days of observation. The 5%, 10%, 15%, and 20%

neem oil shampoo were effective as an anti-head louse. The 15% and 20% neem oil

shampoo had anti head lice effectiveness activity as same as 1% permethrin lotion. In

conclusion, the 15% neem oil shampoo was chosen as the best shampoo, its high anti

head lice effect and light irritation response

Another study conducted on 2018, by the UERM Health Sciences Journal, where

they conducted a study on the effectiveness of neem oil neem seed oil methanolic

extract shampoo versus permethrin shampoo in the reduction of head lice infestation in

children, shows that there was a statistically significant difference in lice count after

treatment for both neem and permethrin, with mean reductions of 17.8 ± 23.97 (p =

0.043) and 22.5 ± 23.47 (p = 0.014), respectively. Repeated Measures ANOVA showed

a summary p-value of 0.041 for neem, 0.013 for permethrin, and 0.193 for the shampoo

alone with a linear trend indicating a significant decrease in the lice counts from the

baseline to the third application of neem and permethrin shampoo, but not in the

shampoo group. There was no significant difference in the mean decrease in lice count

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from baseline to the third application between the neem and permethrin shampoo

groups.

Where in conclusion neem seed oil methanolic extract shampoo is non-inferior

and comparable to permethrin in the reduction of head lice count. There were no

reported dermatologic adverse effects such as burning sensations, redness, skin

irritation, and allergic reactions.

LOCAL LITERATURE

According to Asher, the Neem tree has a wide spectrum of activity which

includes anti-helminthic, anti-feedant, parasiticide, insecticide and pediculocide

properties. The plant’s seed is enclosed by a sweet pulp containing 1-3 kernels, in

which azadirachtin a biologically active compound known to be toxic to insects, is

contained. Azadirachtin is a complex Tetranortiterpenoid limonoid, and this compound

affects both physiologic and behavioral aspects in a wide variety of insects: anti-

feedancy, severe growth reduction, increased mortality, and abnormally delayed molts.

Neem seeds also contain other Triterpenoid components such as 3-tigloylazadirachtin

(azadirachtin B), nimbin, and salannin. These components have an additive effect when

combined with azadirachtin and are useful in preventing resistance.

Schmall also stated that another important effect of neem is stunting in the

development of immature insect stages.15 Due to the reduced food intake by adults,

postembryonic development of species delayed. Neem also causes disruption of the

endocrine system leading to disturbances of molting, pupation, and adult emergence.

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The reproduction of insects is also greatly affected, due to egg sterility and shortening of

the longevity of male and female insects. (UERM Health Sciences Journal, 2018)

In the same article provided by the UERM Health Sciences Journal year 2018, it

was stated that the World Health Organization has also identified Neem tree as an

environmentally powerful natural pesticide for it’s potential use in pest management,

environment, protection, and medicine.

LOCAL STUDIES

One study using the quasi experimental were conducted about the effect of

Azadirachta Indica seed extract on the severity of head lice infestation among selected

children in Barangay Kamputhaw, Cebu City (Arboladura NJM, Arnoco KRV, Arnoco

LGD, Barrios SMD, Borgonia PMAG, Caminong AD, Cayanong LC, Chang JW,

Demafeliz JJC, Ebo CZM, Lo MKN, Pepito JP)

Prior to the actual data gathering, three evaluators, who were chosen by the

group, with 20/20 visions, were taught by Dr. Canete to assess the severity of head lice

infestation. Three evaluators assessed the hair for head lice infestation of each

participant. The evaluators checked the presence of lice and the nits within six

millimeters from the scalp were counted. On day 1 to 5 the extract was applied from the

scalp to the tip of the hair using a paint brush. With gloved hands, the scalp was

massaged for 3-5 minutes and covered with a shower cap for 20 minutes. After 20

minutes, the hair was uncovered, washed and shampooed thoroughly. On day 6, the

three evaluators again examined the hair of each participant following the same

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procedures as with Day 0. Prior to the treatment, the median score on the severity of

head lice infestation obtained was 3 which was categorized as severe infestation. After

the application of the experimental treatment, a median score on the severity of head

lice infestation of 1 was obtained which was categorized as mild infestation. With these,

the null hypothesis was rejected. Thus, there was a significant effect on the severity of

head lice infestation after neem seed extract application.

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 CHAPTER III

RESEARCH METHODOLOGY

This chapter shows the research design, research instruments and materials,

and the following procedure.

RESEARCH DESIGN

Qualitative method will be used in data gathering conducted by the researchers

using the experimental design which allowed them to collect the following data needed

in this research in a given period of time. Quantitative analysis of the Azadirachta Indica

oil extract will be used to determine its efficacy to treat head lice infestation. It will also

be used in determining the amount of extracts needed in order to make a bar shampoo.

Other methods which include the Phytochemical Testing of the efficacy of the product is

expected to be undergone under the quantitative method to be conducted by the

researchers.

INSTRUMENTATION

Collection and Preparation of Materials

 6 tbsp. neem tree seed extract

 ½ cup coconut oil

 ½ cup olive Oil

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  ¼ cup liquid glycerin

 ¾ cup distilled water

 Hand blender

 Pan

 Spoon

 Measuring cup/beaker

 Silicon Molder

 Gloves

 Safety Goggles

 White-clean Cloth

Materials needed in Phytochemical Testing

 Ethanol

 Graduated cylinder

 Flask

 Test tube racks

 Medicine dropper

 Spatula (Ceramic)

 Beaker (250 ml)

 Laboratory heater

 Tongs

 Hot plate

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Other materials

 Distilled water

 Gauze pad

 Empty jar (4 Clean and Sterilized)

 Paper tape

 Marker

 Surgical gloves

 Face mask

 Laboratory gown

 Goggles

PROCEDURE

Preparation and Extraction of Plants

The process will start by the gathering of the seeds of Azadirachta Indica, the

main ingredient and the preparation of the materials to be used. The researchers will

then prepare the neem seeds for grinding. Followed by the extraction of oil from the

mixture of the ground neem seeds and water. After extracting the oil, the other

ingredients together with the main ingredients will be mixed in a bowl to prepare it for

proceeding to the making of bar shampoo. The process will take hours of waiting for the

bar shampoo to be fully molded and ready for packaging.

Phytochemical Screening of Azadirachta Indica

Alkaloid Test

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 Add 2 ml of filtrate to 1 drop of 1% HCI then steam it for 15 minutes, after the

solution is divided into 2 test tubes 1% each. In one test tube, add 6 drops of

dragendorff’s solution and for the other test tube, add 6 drops of Mayer’s reagent.

Creamy participate and brownish red/orange precipitate will indicate the presence of

alkaloid.

Steroid Test

Mix 2 ml of filtrate with 2 ml of acetic anhydride concentrated to sulfuric acid

(H2SO4). A ring formation with the color blue green will indicate the presence of steroid.

Saponin Test

Add 5 ml filtrate with 5 ml distilled water, shake it for about 30 seconds. If 2

inches of “honey combed” froth is formed and stay for 10 minutes, the test subject is

positive for saponin.

Tannin Test

Add 3 ml of filtrate to 2 ml of FeC13, Blue-black precipitate will indicate the

presence of tannin.

Cardiac Glycoside Test

Add 2 ml of filtrated Azadirachta seeds with 1 ml of glacial acetic acid, 1 ml of

FeC13 and 1 ml of concentrated Sulphur acid (H2SO4). Green-blue color will indicate

the presence of cardiac glycoside.

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Flavonoid Test

Mix 2 ml of filtrate with 2 ml concentrated hydrochloric acid (HCI) and 4 strips of

magnesium. Tomato red color will indicate the presence of flavonoids.

NEEM SEED OIL EXTRACTION PROCESS

1. Gather the main ingredients which is the seeds of the neem tree (Azadirachta

Indica).

2. Prepare the materials needed for the extraction of the seed oil.

3. After the preparation of the ingredients and materials, start by washing the seeds

to make sure that it is thoroughly cleaned.

4. Blend the seeds of the neem using a blender.

5. Put the ground neem seeds into a bowl and cover it with water.

6. Stir it once and wait for the oil to start flowing upon the water.

7. Skim the oil and put it in a container.

PROCEDURE IN MAKING THE BAR SHAMPOO

1. Get a clean bowl and mix the 6 tablespoon of neem oil extracts with ½ cup of

coconut oil, ½ cup of olive oil, ½ cup lavender oil, ¾ cup of cool distilled water,

and ¼ cup of liquid glycerin.

2. After putting all the ingredients, mix it constantly with a hand blender.

3. Put the mixture into a silicon molder and let it cool.

4. Wait for some hours until the bar shampoo is ready and fully molded.

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