Chapter 1

PROBLEM AND ITS SCOPE

Background of the Studies

In the Philippines most family households has pet dogs in them. And mostly in those dogs
are living ticks that sucks blood in the dogs. The skin where ticks attach to humans can become
red and irritated. Ticks that transmit diseases to your dog can also transmit many of the
same diseases to people. ... Diseases, such as Lyme disease and Rocky Mountain spotted fever,
which have already been described in dogs, can also be very serious in humans. It could be
dangerous to those children or adults who come in contact with theqdir dogs. We used atis leaves
because it could be found mostly anywhere around in our area and it is free from those
dangerous chemicals found in other insecticides. We also used atis leaves because of its
composition making it insecticidal. The extract of the atis leaves can be used as a killer substance
to dog ticks more specific Dermacentor variabilis. 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, 2010).

Statement of the Problem

Diseases that comes from dog ticks can be dangerous to our healths. The researchers goal is to
make an alternative ixodicide product using Annona Squamosa leaves extract.
Specifically, this study aims to answer this following questions.
1. Will the product be effective on the subject?
2. Would there be other effects on the subject?
3. Will there be other function for the product?
Statement of Hypothesis
The researchers thought that by spraying the Annona Squamosa leaves extract to the subject,
the subject would be killed because of the chemical in the extract.
Significance of the Study
To these, Filipino households that has pet dogs in them are the main beneficiaries. This study
helps them from possible bites of dog ticks that might transfer from their dogs to them, and
possible prevention of transfer of tick borne diseases. The dog themselves can benefit from this
study for it would remove the ticks that sucks their blood and prevent diseases that could occur
to the pet dogs or stray dogs.
Scope and Limitation
This study only limits in using atis leaves extract as dog tick killer.

Definition of Terms
To fully understand the content of the study, here are the terms defined.

Dog Tick – Insect used as test subject for the alternative dog tick killer.
Atis Leaves – The material that is going to be extracted.
Ethanol – Helps in bringing out the atis leaves extract.
Anthraquinones – The chemical that will be extracted from the atis leaves.

Theoretical Background

Chapter 2
Review of Related Studies and Literature

Related Literature
Atis (Annona squamosa ) known in English as sugar apple or sweetsop, a member of plantae
Kingdom and is the most widely grown species of Annona. Studies show that its fruit, leaves and seeds
contains compositions that makes them vermicidal and insecticidal. It is native to the tropical Americas
and West Indies and the Spanish traders of the Manila galleons were the ones who brought it to Asia,
where its old Mexican name ate may still be found in Bengali ata, Nepalese aati, Sinhalese mati anoda,
Burmese awzar thee, Indonesia “ Srikaya”’ and atis in the Philippines. It is also known as Seetaphal in
India and Shareefa Pakistan and in the Philippines and in Australia. The name is also used in Portuguese
as atan.

Dermacentor variabilis is a blood sucking insect but bigger than lice, it is known to carry
bacteria responsible for several diseases in humans and animals. Depending on what species its
lifecycle varies. Most ticks go through four stages: egg, six-legged larva, eight-legged nymph,
and adult.Ticks that are feeding are the ones who transmits pathogen to the host, preparing to
feed can take from 10 minutes to 2 hours. When the tick found a spot, it sticks to it, cut the
surface and insert its feeding tube. Some species even secrete a cement-like substance to be
firmly attached during its feeding time. It also secretes a small amount of saliva with anesthetic
properties making the host not 9noticing the tick. If the host animal has a blood borne disease
then the tick will ingest the pathogens with the blood. Ticks can also enter a small amount of its
saliva to the host animal and if the tick contains pathogen, the organism may be transmitted in
the process. After feeding the tick mostly drops off from its host and prepare for its next life
stage where it could transmit an acquired diseases to its new host. They can also live up to 2
years without a host.

Related Local Study

According to Ballesteros et al (2014) the Annona Squamosa leaves extract is found to have
pediculicidal effect potential against lice found within the Philippines. Their results show it is
less effective compared with the commercially prepared Licealiz for they have observed that in
just a span of four hours the mortality of the of lice increased but the treatment with the fastest
pediculicidal effect was with Licealiz. They concluded that the habitat of the lice is also a major
factor, food also is an important factor because lice dies in just a span of four hours with or
without treatment.

Related Foreign Study

According to Lary C. (2018) that the Apple Cider Vinegar (Malus pumila Mill) is a natural
solution for tick prevention and there are multiple ways to use it to your dogs. She concluded that
the Mallus pumila Mill contains insecticidal properties that removes ticks from the dog but
doesn’t kill it. The substance alone cannot kill the tick for ticks, so it is combined with neem oil
and water, mix it and spray to the ticks and the results will be seen after.
Jaswanth A. et al (2001) stated in their study that the methanolic extract of A.squamosa leaves
contains results that suggest the potential mosquitocidal effect of A.squamosa against
C.quinquefasciatus. The leaves were collected in the fruit seasoning of September 2000, then the
leaves were shade dried, pulverized by a mechanical grinder, sieved through 40 mesh. The
powdered leaves then were extracted with methanol using soxhlet extraction apparatuses and
then its was concentrated and dried under reduced pressure. The substance then was dissolved in
the deodorized kerosene and filtered using Whatman filter paper No.1 and stored in a well closed
container. The extract was then called liquid mosquito insecticide extract formulation (LMIEF).
 Chapter 3
Methodology

A. Collection of the Materials

Here are the materials used for this study, there were materials that were collected and others
were bought. Here are the list:
 Atis Leaves
 Dog Ticks
 Mortar & Pestle
 Funnel
 Filter Paper
 Ethanol
 Measuring Spoon (5mL)

B. Preparation of the Materials

1. Atis Leaves Preparation
The atis were washed thoroughly to remove unnecessary dirt and any other substance that
might affect the process. The leaves were soaked in distilled water for about 12 hours.

2. Dog Tick Preparation

The ticks are freshly collected from the dogs for them to have eaten before the testing.They
are contained on different kinds of containers.

C. Extraction Proccess
The atis leaves will be pounded thoroughly and will be soaked in a cup filled with ethanol
for one hour. After an hour the ethanol containing the leaves will be filtered using filter paper.
The filtered extract will undergo in a process called water bathing, by the time it reaches its
boiling point which is 78.38 degrees celcius the extract would be filtered again in a new piece of
filter paper. After filtering you will get the atis leaves extract.

D. Preparation of the Extract

The extract then is going to be divided into three in preparation for the three trials. The first
contains 10 mL, the second one contains 20 mL and the third one contains 30mL.

E. Isolation Process
The ticks will be divided into 3 containers. First one contains 3 ticks, second one contains 6
ticks and third one contains 9 ticks.

F. Experimentation
The atis leaves extract are placed unto three various measurements; 10mL, 20mL, 30mL.
There would be three sets of testing to compare each results of each sets. The extracts will
then be applied to each containers.

Schematic Diagram

COLLECTION OF SCIENCE

PREPARATION OF THE
MATERIALS

EXTRACTION PROCESS

PREPARATION OF THE
EXTRACT

ISOLATION PROCESS

EXPERIMENTATION
 Chapter 4
Presentation, Analysis and Interpretation of Data
The following tables shows the results of the researchers experiment.

A. Results of the Experimentation

Table 1. Set 1 (10 mL)
Test Subjects Size Time of death
(minute)
Tick 1 Small 4 min
Tick 2 Medium none
Tick 3 Large none

Table 1 shows the result of experimentation of set 1 using the alternative ixodicide on the
dog ticks, the ticks that was labeled as none, were the ticks that didn’t die in the experiment.
However, base on the researchers observation these two ticks at first stopped moving at about 5
minutes and concluded that its was dead but after another 5 minutes the ticks were moving again
except for the small one.

Table 2. Set 2 (20mL)

Time of death
Test Subjects Size (minute)
 3 min
Tick 1 Small
6 min
Tick 2 Small
none
Tick 3 Medium
none
Tick 4 Medium
none
Tick 5 Large
none
Tick 6 Large

In Table 2, it shows the results of the second test done by the researchers to have more
results and compare them, the set 2 results shows that the small sized ticks are the only ones that
was only affected by the product.

Table 3. Set 3 (30mL)

Time of death
Test Subjects Size (minute)
Tick 1 Small 3 min

Tick 2 Small 2 min

Tick 3 Small 6 min

Tick 4 Medium none

Tick 5 Medium none

Tick 6 Medium none

Tick 7 Large none

Tick 8 Large none

Tick 9 Large none

 Table 3 shows that only the small sized ticks are only the ones that was taken effect by the
product of the researcher.

Discussion
As shown in these tables, the researchers had stated their results base on their observation
and analysis. As the researchers had presented in Table 1, they had conducted the first set
without the proper place and materials so the results might be inaccurate so they still put the
results base on their observation. After the first set was done the researcher gathered ticks again
and conducted experimentation again at the researchers house with the proper materials and
stated the results base on the observation, seeing the results the only ones that are affected by the
extract were the only small sized ticks and the effect would take effect in just a span of two to six
minutes and the results would be seen after the time.

Chapter 5
Summary, Conclusion and Recommendation

Summary of the Study

In the Philippines, pet dogs are common on family households and chances are these dogs
are infested with ticks that might carry diseases that would be dangerous to both human and
dogs. So the researchers conducted a study whether the Annona Squamosa leaves extract would
work as an alternative ixodicide. The researchers had gathered the materials and conducted a test
on the subjects, after the test they presented their results based on their observation. Results had
shown that even though the ticks are difficult to kill the extract had taken effect but to the only
small sized ticks only. It also observed that all the ticks stops moving when sprayed by the
product but later on the ticks moves except for the small ones.

Conclusion
 The researchers concluded that the extract of the atis (Annona Squamosa) leaves
containing the chemical Anthraquinones were effective against small sized dog ticks only
therefore it could be used as an alternative ixodicide against dog ticks.

Recommendations
Since the aim of the study is to test whether the product would work against the test
subjects. The researchers would like to recommend the following:
 To use the other parts of the atis such as seeds and fruit skin.
 To test whether
 To add another variable to the product to determine if there is an increase on its
effectiveness on the test subject.

REFERENCE