UNIVERSITY OF THE EAST

SENIOR HIGHSCHOOL- SCIENCE AND TECHNOLOGY, ENGINEERING &

MATHEMATICS

A Comparative Analysis in the Effectivity of Coffea arabica (Arabic

coffee) and Cyanobacteria (Blue-green Algae) as an Organic
Larvicide Against Musca domestica (House Fly) Bred Through
Rearing Method

SUBMITTED BY:
GROUP 5- STL12-2P

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 CHAPTER 1
INTRODUCTION

I. Background and Rationale of the Study

The common housefly, Musca domestica, is one of the most prevalent species of
insects in the world. This type of insect feeds on various things such as food scraps,
and wastes. They also live in dense areas as well as breed in feces or garbage. Under
these conditions, they are well-known to be carriers of various diseases and is a
known health-hazard among those who have poor sanitary conditions.

130 pathogens were identified in an average house fly in which most are bacteria,
including the life-threatening kind. This can cause serious infections among humans
and animals. (Khamesipour, F., et. Al 2018) Due to this becoming a nuisance, various
researches have advanced to innovate different ways on how to eliminate these kinds
of insects.

It is known that in some plants, caffeine acts as natural pesticide that enables the
paralysis of insect or at times, even death. In a study conducted by Bicudo, Jose, and
Theodoro (2003) it is said that caffeine and coffee grounds were able to block the
growth of larvae from the species Aedes aegypti. Lower concentration levels resulted
in more advanced stages in the growth of larvae which implies that high
concentration levels of coffee grounds have significant effect on the growth of larvae.

Limited studies have also looked into Cyanobacteria, known as Blue-green algae,
to contain toxins that are able to kill larvae. (Marten, G. 2007) It is said that because
of its toxic components, in higher concentration and quantity, it may offer
possibilities as larvicides.
 This study aims to ascertain the effectivity of both Coffea arabica, and
Cyanobacteria as organic larvicides in different concentrations as it proposes better
alternative options in the elimination of insects, specifically Musca domestica.

II. Research Objectives

The research aims to test the effectivity of Arabic Coffee (Coffea arabica) and
Blue-green algae (Cyanobacteria) against the larvae of a common housefly (Musca
domestica). Specific objectives that the researchers aim for are the following:

1. To measure the difference in mortality rate caused by Arabic coffee and Blue-
Green algae.

2. To know which organic larvicide is much more effective against Musca

domestica.

3. To find the concentration level at which Arabic coffee is most effective against
the larvae of Musca domestica.

4. To find the concentration level at which Blue-green algae is most effective against
the larvae of Musca domestica.

5. To compare how much the concentration changes affects the effectivity of Coffea
arabica and Cyanobacteria against Musca domestica.

III. Research Hypothesis

I. Null Hypothesis

There is no significant difference on the mortality rate of the fly larvae between
the 5 test experiments. The 5 test subjects showed almost the same (if not the same)
mortality rates after applying both organic larvicides.

II. Alternative hypothesis

 There is a significant difference in the mortality rate of fly larvae between the 5
test subjects. The mortality rates varied on the type of organic larvicide used in each test
subjects.

• The mortality rate of fly larvae was higher when the concentrated Arabic coffee
was applied on test subjects.

• The mortality rate of fly larvae was higher when the concentrated blue-green
algae was applied on the test subjects.

IV. Conceptual Framework

Rearing Method
(supported by
IRAC)

Coffea arabica

Effectivity based
To
Musca domestica on the mortality
rate

Cyanobacteria

Analyzed using
 Caffeine is the most common drug that human intake. Although there are
numerous positive reasons as to why it is regularly taken, there are also adverse health
effects unbeknownst to some. Since caffeine is a central nervous system inhibitor, one of
the aspects it affects is locomotion. According to Barry D. Smith, Uma Gupta, and B. S.
Gupta (p. 47, 2007), low doses of caffeine increases locomotion activity on rats but
decrease it with high concentrations. A remarkable example of it is the study conducted
by DR. James Nathanson who observed that insects displayed a change in behaviour and
growth when exposed to caffeine. Their Caffeine and Activation theory will be used in
the current study, to provide explanations on the effects of caffeine in house flies during
their larval stage and how it affects their behaviour or induces death.

In determining, the relationship between the stimuli (dose) to the mortality rates
of biological organisms- in this case the blue-green algae and coffee beans- the
researchers will use Probit Analysis. Probit analysis is a statistical treatment that is
applicable to deduce the response of toxicity levels of substances to the dependent
variables. Sparling (2016), described in his book ‘Ecotoxicology Essentials’ that “Probit
is well-suited for characterizing binomial response variables (e.g., live or dead, diseased
or healthy) such as those derived from dose–response experiments.” This will simplify
the data as it will be presented in a linear form (Figure 1) to be followed by a regression
analysis.

Fig. 1: Probit Analysis (https://www.sciencedirect.com/topics/agricultural-

and-biological-sciences/probit-analysis
V. Significance of the Study

The findings of this study aims to identify the effectivity between Coffea arabica
(Arabic Coffee) and Cyanobacteria (blue-green algae) as organic larvicide against Musca
domestica (housefly), as it will benefit the following:

1. Community. Areas which have poor sanitary conditions have high probabilities
of infestations from insects such as Musca domestica which may greatly affect those who
are residing there. The study’s goal is to find an effective alternative organic larvicide
which may be utilized by different communities.

2. Environment. The environment in which we live in is greatly affected by

numerous substances and chemicals that are being used to fight off pests. The research
will be able to determine whether there are much more effective alternative options that
are safer, and cheaper to use.

3. Farmers. One of the main concerns of farmers especially those who are in
poultry

4. Future researchers. Researchers who will be focusing on this field may be able
to obtain additional knowledge about the mortality rate of insects such as Musca
domestica in organic variables such as Cofea arabica and Cyanobacteria.

VI. Scope and Delimitation of the Study

 The main focus of this study is to identify and compare the mortality rate between
Coffea arabica (Arabic coffee) and Cyanobacteria (Blue-green algae) as organic
larvicides against the larvaes of Musca domestica (Housefly). The researchers will be
utilizing the method of breeding through rearing in obtaining samples of larvae from the
houseflies. The researches shall be limiting the variables to Coffea Arabica and
Cyanobacteria as the organic larvicides, while this is limited as well to the species of
Musca domestica as the subject. As this study is only limited to houseflies, the obtained
data will not be applicable to other insects.

VII. Definition of Terms

For a better understanding of the study, the following terms are defined in the
context of this research.

1. Coffea arabica. It refers to the scientific name for Arabica Coffee that makes the
organic larvicide.
2. Cyanobacteria. It is the scientific name for Blue-green algae that makes the
organic larvicide along with the Arabica Coffee
3. Musca domestica. It refers to the scientific name of housefly.
4. Rearing Method. It is the process or method that will use on raising house flies.
 CHAPTER II
REVIEW OF RELATED LITERATURE
In this chapter, literature and studies that are associated with the topics and provides an
overview with the present study are thoroughly discussed. This includes journals, articles, and
excerpts from published & unpublished studies and books.

A. Larvicidal potential of five Philippine plants against Aedes aegypti (Linneus) and
Culex quinquefasciatu (Say). Southeast Asian J Trop Med Public Health.

Anona squarnosa (atis), Azadirachta indica (neem) and Codiaeurn variegatem

(Croton) Eucalyptus globulus (olive gum eucalyptus), Lansium dornesticum (lansones),
are all used in a study conducted by Monzon, Alvior, Luczon, Morales, and Mutuc
(1994). Their study aimed to use different plants found in the Philippines noted by their
insecticidal properties as an organic alternative insecticide. They subjected two species of
mosquitos namely Aedes aegypti and Culex quinquefasciatus to varying concentrations
of the five plants then used Probit Analysis for analysing the results. They discovered that
Lansones is the most potent against Aedes aegypti while Atis is more effective against
Culex quinquefasciatus. The variation is said to be because of the level of toxicity present
in the plants as well as the mosquito’s physiological properties.

B. The larvicidal activity of brown algae Padina minor and Dicyota linearis against the
dengue vector, Aedes aegypti

A research was conducted in 2013 by Bantoto and Dy about brown algae and it’s
larvicidal effects on Aedes aegypti. The brown algae solution used were tested on
different concentrations; 20 mg/ml, 40 mg/ml, 60 mg/ml, 80 mg/ml, and 100 mg/ml. At
 20% concentration, the Padina minor showed 8% mortality rate while the Dicyota linearis
showed zero. At 40% -80%, the mortality rates rose with Padis minor consistently being
higher. At 100% both algal extracts showed 100 percent mortality rates. The study shows
that varieties of algae are effective as larvicidal agents and can be used as alternative to
chemical larvicides.
C. The larvicidal effect of used ground coffee (Coffea) Aedes mosquito larvae

Bakrie, et al. (2017) conducted a research on the larvicidal effect of coffee on

fourth instar (Aedes) mosquito larvae. The researchers used a concentration of 1% and
5% of coffee and are compared to baygon (positive control) with the same concentration.
These concentrates are applied on 175 mosquito larvae. The results showed that at 1%
concentration, there is a 43% mortality rate on the mosquito larvae. As for the 5%
concentrate, it resulted to 63% morality rate. On the other hand, the experiments where
baygon is applied showed 100% mortaliy rate. The study shows that coffee is already
potent at 5% concentration and could be more effective when used on higher
concentrations

D. The effectiveness of arabica coffee (Coffea arabica L) grounds on mortality and

growth of Aedes aegypti Larva

Aditama, Zulfikar, and Sitepu (2016) conducted a research on Coffea arabica as a

larvicide against Aedes aegypti . The study made use of ground coffee as an inhibitor on
the growth and as a causal agent on the mortality of Aedes aegypti larvae. The mosquito
larvae were exposed to different doses of Coffea Arabica. The mortality and growth data
were then analysed by specific methods. The results of the study showed that the
mortality rates of the Aedes aegypti larvae ranged from 6.67%-76.67% varying from the
levels of concentration of Coffea arabica. The results also showed a decrease in the
growth index (GI) value of the larvae, which shows the hindrance on the growth of the
larvae.

E. Mosquito larvicide from Cyanobacteria

 Berry (2014) wrote a dissertation about Cyanobacteria as a larvicide against
Aedes aegypti. The study tested different types of lower forms of Cyanobacteria. One-
hundred twenty (120) test samples were made and are dosed with varieties of
Cyanobacteria concentrated on different solvents. The polar extracts of the Cyanobacteria
samples were separated and mixed with varieties of solvents and applied to the test
subjects. The experiment results showed that out of 120 experiments, 31 showed twenty-
five percent (25%) or more mortality rates. This shows that Cyanobacteria is an effective
larvicide against Aedes aegypti even when concentrated on different solvents.

F. Coffee and its waste repel gravid Aedes albopictus females and inhibit the
development of their embryos

Satho, et al. (2015) studied about the effects of coffee on Aedes albopictus
females on their oviposition and larval eclosion on different concentrations. The
experiment used Coffea canephora that was dissolved on water on different
concentrations. The mosquitoes were placed in an environment where they were allowed
only to breed in cups containing water and the others containing coffee on different
concentrations. The tests results showed that the mosquitoes preferred laying eggs on
water containing cups. The amount of eggs laid on a cup containing coffee only ranged
from 3% - 11% varying on levels of coffee concentration, as the mosquitoes preferred to
lay on water containing cups.

G. Toxicity
Due to the growing concern of insecticide resistance and the search for an
insecticide that does not negatively affect non – targets, researchers have been trying to
find an alternative organic insecticide that affects pests or insects during their larval
stage. Phankaen et al. (2017) studied how the extracts of Coffea arabica affected
Tribolium castaneum (Habst) under laboratory conditions. Hexane, dichloromethane,
ethyl acetate, and methanol are the four solvents used along with the coffee extract. They
 found out that caffeine extract showed very strong repellency on olfactometer. Fumigant
toxicity test also showed that dichloromethane was more toxic compared to others.

H. Algae
Seven out of 20 marine algae found in South West Coast of India, proved to have
the capacity to be a possible organic substitute to chemical insecticides. The algaes were
extracted in methanol and were tested against Culex quinquefasciatus (Say) and Aaedes
aegypti (Linnaeus) during the third to fourth instar larvae stage. Lobophora variegate is
the most potent among the seven. Padina tetrastomatica exhibited the least larvicidal
activity. Maninal et al. (2011) aimed to compare the potential of the algaes to prevent the
growth of mosquito and say.

I. In vitro Mosquito Larvicidal Activity of Marine Algae Against the Human Vectors,
Culex quinquefasciatus (Say) and Aedes aegypti (Linnaeus) (Diptera: Cullicidae)

The study of Manilal et al. (2006) focuses on the larvicidal effect of different
marine algae against the human vectors Southern house mosquito, and Housefly. The
main objective of this study is to compare the ability of organic extracts of marine algae
against the two human vectors. Twenty marine algae were collected from the southeast
coast of India and extracted it in methanol. Out of the twenty marine algae collected,
early stage larvae were found to be very sensitive in seven of them. After comparing the
larvicidal potency among the seven marine algae, Padina tetrastromatica ranked highest
in larvicidal efficacy. In conclusion, this marine algae is found as a potential resource of
natural insecticide that can be developed to replace inorganic insecticides in the future.

J.
 CHAPTER III
MATERIALS AND METHODS

A. Materials

The researchers shall be utilizing a variety of materials in the making of the

research. The independent variables will be the Arabic coffee and cultured Blue-green
algae. Larvae will also be harvested from mature houseflies.

For the rearing procedure, a large storage box shall be used to create a controlled
environment and safe space where live houseflies may be able to thrive and breed. The
bait that will be utilized are food scraps such as rotten fish which are known baits.

For the experiment itself, tools such as forceps will be utilized that will be needed
in transferring specimens to different containers. A beaker shall also be utilized for
accurate measurements in the concentration of the independent variables. Additional test
tubes as well as test tube racks may be needed in order to contain excess concentrated
variables. A dropper shall also be utilized once the solutions are ready to be transferred to
the larvae.

For the containers where the controlled variables as well as dependent variables
will be placed, a plastic jar which amounts to 450ml will be used. Cheese cloth shall be
 utilized in order for the houseflies to be contained once there will be test subjects who
will be able to fully develop in the adult stage under the given circumstances.

B. Procedure

C. Schematic Diagram
Breeding Musca
domestica through
Rearing method

Extract the Coffea arabica

and Cyanobacteria

Collect the larvae then

transfer it into a different
container

Container Container Container Container Container Container Container

A B (1) B (2) B (3) C (1) C (2) C (3)

Controlled Coffea arabica with Cyanobacteria with varying

Group varying concentrations will concentrations will be
be exposed to houseflies in exposed to houseflies in
these containers these containers