Kaunlaran High School

RESEARCH PLAN

Title

Assessment of affectivity of Gambusia affinis as biological control for Aedes aegypti larvae in
artificial fish tank installed in Kaunlaran High School

A. RATIONALE

Dengue is fast emerging pandemic-prone viral disease in many parts of the world. Dengue
flourishes in urban poor areas, suburbs and the countryside but also affects more affluent
neighborhoods in tropical and subtropical countries. Dengue is a mosquito-borne viral infection
causing a severe flu-like illness and, sometimes causing a potentially lethal complication called
severe dengue. The incidence of dengue has increased 30-fold over the last 50 years. Up to 50-
100 million infections are now estimated to occur annually in over 100 endemic countries, putting
almost half of the world’s population at risk. Severe dengue (previously known as dengue
haemorrhagic fever) was first recognized in the 1950s during dengue epidemics in the Philippines
and Thailand. Today it affects Asian and Latin American countries and has become a leading
cause of hospitalization and death among children and adults in these regions. The full life cycle
of dengue fever virus involves the role of mosquito as a transmitter (or vector) and humans as the
main victim and source of infection. (“Dengue and severe dengue.” World Health Organization,
2019, https://www.who.int/news-room/fact-sheets/detail/dengue-and-severe-dengue)

Elimination of mosquito has been tried by various methods, however, due to certain limitations,
related with environmental issues; there is a shift in approach to control the mosquitoes
population. Due to ban or restrictions by environmental protection agencies, there are now fewer
chemicals available than there have been for the last 20 years (Walker et al., 2002).

Mosquito control has been a point of focus in scientific communities, especially health-providing
units. Various methods have been tried and many more still being developed to eliminate or
reduce mosquito populace. Certain environmental issues related with chemical control of
mosquitoes have convinced experts to opt for eco-friendly methods. Among those methods are
the biological control methods, which suppress vector populations through
introduction/manipulation of organisms (natural enemies, e.g., parasites, disease organisms and
predatory animals). Present study was focused on using edible fish as predator of mosquito
larvae. Experimental results revealed that, edible fish have tremendous potential to be used as
larvivorous predator of mosquito. It was further found that, larvivorous fish (Gambusia Affinis),
 used in this study preferred live larvae when compared with commercial food. (Noreem M.
et al. “Biological control of Mosquito larvae using edible fish.” Department of Veterinary
Parasitology, Sindh Agriculture University Tandojam-Pakistan.
https://www.researchgate.net/publication/324439144_BIOLOGICAL _CONTROL
_OF_MOSQUITO_LARVAE_USING_EDIBLE_FISH

Concerns have also been felt about introducing exotic fish for mosquito control due to their possible
impact on native fish species. The introduction of Gambusia affinis in Greece (Europe) led to a
decline of the endemic fish species Valencia letourneuxi (Hoddle, 2004) and similar findings were
reported in United States, Spain and Australia (Economidis, 1995).

In the last few decades, conservationists have increasingly called for action to protect native animal
species and habitats from ‘‘biological invasions’’ of invasive alien species. The presence of animals
and plants that adversely affect both the ecological integrity and the local economy in areas in
which they are not indigenous is common across the globe. Their dispersal is often due to voluntary
and involuntary human activities. The negative impact of invasive alien species on biodiversity is
second only to habitat loss. Mosquitofish (Gambusia affinis) is an example of a freshwater fish
species whose remarkable diffusion outside its native range has led to it being placed on the list of
the world’s hundred worst invasive alien species (International Union for Conservation of Nature).
Biological invasions rank among the main global threats for biodiversity. (Polverino et al.
“Mosquitofish (Gambusia affinis) Preference and Behavioral Response to Animated Images of
Conspecifics Altered in Their Color, Aspect Ratio, and Swimming Depth.” Department of
Mechanical and Aerospace Engineering, Polytechnic Institute of New York University, Brooklyn,
New York, United States of America.
http://sci-hub.tw/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3546983/
Background
of the Study Dengue illness is the most important mosquito-borne viral disease in the world. It is
(What is the
current epidemic in the Philippines and considered one of its eight pervasive infectious diseases.
situation, Recently, Department of Health requested the National Disaster Risk Reduction and
what is your Management Council to convene a declaration of national dengue epidemic in the wake
sample, cite
sources of the 146,062 cases recorded since January up to July 20, 2019, 98% higher than the
generously) same period in 2018 that result to 622 deaths. Based on the DOH Dengue Surveillance
Report, Region VI (Western Visayas) had the most number of cases at 23,330, followed
by Region IV-A (CALABARZON) with 16,515, Region IX (Zamboanga Peninsula) with
12,317, Region X (Northern Mindanao) with 11,455, and Region XII (SOCCSKSARGEN)
with 11,083 cases. Creating different strategies in controlling the populace of Mosquito
that carry dengue virus (DEVN inclined due to the rise of Dengue incidents. Current
mosquito control techniques implemented involve larval source reduction, larviciding, and
space spray of adulticides which are costly, laborious, and of debatable efficacy. Due to
different issues in chemical usage in eradicating Mosquito-borne diseases, Biological
control was introduce. Biological control, particularly using larvivorous fish, was important
to malaria control programmes in the 20th century, particularly in urban and periurban
areas for immediate use in developed and developing countries . It has a very positive
role to play in the integrated control methodologies in which both pesticides and fish or
other biotic agents have their own roles. Biological control refers to the introduction or
manipulation of organisms to suppress vector populations. A wide range of organisms
helps to regulate mosquito populations naturally through predation, parasitism and
competition. As biological mosquito control agents, larvivorous fish (i.e., those that feed
on immature stages of mosquitoes) are being used extensively all over the world since
the early 1900s.

Mosquito Fish (Gambusia affinis), a fresh water fish species was one of biological
control for mosquito due to its diet is predominantly consists of large amounts of
mosquito larvae and can also survive in waters with low oxygen purposely introduced
from its native Texas (Southern USA) to the Hawaiin Islands in 1905 . In 1921, it was
introduced in Spain; then from there in Italy during 1920s and later to 60 other countries
including the Philippines. However, recent studies prove that Mosquito fish is an invasive
freshwater fish. A group of Spanish researchers has analysed, the evolutionary changes
of this animal through its genes. According to the study, genetic variability has allowed
this fish to adapt and spread throughout its new environment. Havoc can be wreaked on
native ecosystems when a foreign species is introduced, becomes acclimated and
expands. Biological invasions are one of the main global threats to biodiversity and, for
that reason, the subject of careful study. The Eastern gambusia (Gambusia holbrooki),
also known as the mosquitofish, was taken from the United States to Spain in the 1920s
as a way to naturally control disease-carrying mosquitoes, such as the ones carrying
malaria. This animal, however, has had an enormous environmental impact on local
fauna, thus becoming one of the world's 100 most harmful invasive exotic species,
according to the International Union for Conservation of Nature (IUCN).

Based on diet, mosquitofish are classified as larvivorous fish. Their diet consists

of zooplankton, small insects and insect larvae, and detritus material. Mosquitofish feed
on mosquito larvae at all stages of life. Adult females can consume hundreds of
mosquito larvae in one day. Maximum consumption rate in a day by one mosquitofish
has been observed to be from 42%–167% of its own body weight. They can suffer
mortalities if fed only on mosquito larvae, and survivors show poor growth and
maturation Mosquitofish have also shown cannibalistic behavior in laboratory
 experiments; however, whether these traits are hereditary is unknown

Mosquitofish were intentionally introduced in many areas with large mosquito

populations to decrease the population of mosquitoes by eating the mosquito
larvae. However, most introductions were ill-advised; in most cases native fish had
already proven to supply maximal control of mosquito population and introducing
mosquitofish has been more harmful to indigenous aquatic life than to the mosquito
population. Introductions outside the mosquitofish's natural range can be harmful to the
nonnative ecosystems. Mosquitofish have been known to kill or injure other small fish by
their aggressive behavior and otherwise harm them through competition. They are now
considered just slightly better at eating mosquitoes than at destroying other aquatic
species. The ecological impacts of mosquitofish are partly dictated by their sex ratio,
which can vary dramatically across their introduced range. Mosquito fish in Australia are
considered noxious pests where they pose a threat to native fish and frog populations
and no evidence indicates they have controlled mosquito populations or mosquito-borne
diseases.

The study will be beneficial to the following;

1. The study will create a method of using Mosquito fish (Gambusia affinis) as
biological control for Aedes aegypti without harming the indigenous aquatic life in
Significance
fresh water ecosystem and at the same time not harming the wellbeing of used
of the Study
(What is the Mosquito fish (Gambusia affinis)
importance of 2. The study will assess the affectivity of using Mosquito fish (Gambusia affinis) as
your research
biological control for Aedes aegypti larvae
based on
your 3. It will provide a design plan for creating artificial fish tank where the Mosquito fish
background?) (Gambusia affinis) can live while used as biological control for Aedes aegypti
4. The study will contribute in lessening the use of pesticides and larvacides which

hazardous to human health and other living things

Brief Specially, the research sought answers to the following questions:

statement of
the problem 1. How effective to use Mosquito fish (Gambusia affinis) as biological control for
Aedes aegypti that caries dengue virus (DENV)
2. Does the Mosquito fish (Gambusia affinis) can adopt in the artificial fish tank
3. How will the study benefited the Government mosquito control programs
implemented to lessen or eliminate the mosquito that carries Dengue Virus
(DENV) in terms of:
 a. Effectiveness;
b. Affordability; and
c. Time management

RESEARCH QUESTION(S), HYPOTHESIS(ES), ENGINEERING GOAL(S), EXPECTED

OUTCOMES
RESEARCH Is there a significant positive effect of using Mosquito fish (Gambusia affinis) as
PROBLEM Biological Control of Aedes aegypti larvae in Kaunlaran High School?
(Declarative or
Interrogative)

GOALS
(Enumerate The research sought to answer the effectiveness of using Mosquito fish (Gambusia
your targets
affinis) as Biological Control of Aedes aegypti larvae in Kaunlaran High school
as you do your
research)

There is a significant positive effect of using Mosquito fish (Gambusia affinis) as a

EXPECTED
OUTCOMES Biological Control of Aedes aegypti larvae in Kaunlaran High School

Null Hypothesis:
There is no significant effect of using Gambusia affinis as a Biological Control of Aedes
HYPOTHESES aegypti larvae in Kaunlaran High School
(Null and
Alternative or Alternative Hypothesis:
the If… then…) There a significant positive effect of using Mosquito fish (Gambusia affinis) as Biological
Control of Aedes aegypti larvae in Kaunlaran High School

Experimental Design
METHODS
Procedure:

1. Gathering of Materials
Researchers will collect the needed part for the fish tank and will look for 10
local female Mosquito fish (Gambusia affinis), also known as kataba. The
gender of Mosquito fish must be analyze carefully to avoid fertilization and
spread of Mosquito fish population in the fish tank. Using female Mosquito fish is
more effective than the male since it can eat hundreds of larvae a day and
easily identify due to its difference of appearance from the male Mosquito fish

2. Constructing and installing the fish tank

The fish tank will be place at the back of mate-shift classroom since the place is
shady, where mosquito usually stay. The fish tank will be near the garden so
 that when the water of the fish tank replace it can be used to water the plant in
the garden and also mosquitos typically want place with lots of plants and
vegetation. There should be no cover in the tank and will place 3 stick of
lawanet for mosquitos to lay their eggs. The solar panel will install at the top of
the roof where it can reach the sunlight. The alternative power supply for aerator
will be place away from the tank that cannot be reach by the water

3. Installing the Mini V380 1080P IP Camera

The V380 1080P IP Camera will be installed at the side of the fish tank and will
stick a small led light at side of the fish tank so that the Cam would be able to
record clearly

4. Realising Mosquito fish (Gambusia affinis) in the fish tank

10 local female Mosquito fish (Gambusia affinis) will be release in the fish tank.
The researcher will check regularly if the mosquito fish where able to fed larvae.
The researcher will give feeds or fish food twice a day so that they can still
survive even there is no larvae hatched from lawanet stick
5. Gathering data
a. The researcher will analyzing the data recorded by Mini V380 1080P IP
Camera
b. The researcher will gathered information about how many dengue cases
occurs from the previous month in the classrooms near vicinity where the
fish tank will be installed. After a month of using the fish tank the reasercher
will check the number of dengue cases occurs.
c. The researcher will provide quetionaires for the students in the classroom
near the place where the fish tank will be installed. The quetionaire tend to
ask the students about how serious the mosquito in the area, this will be
done before installation and a month after installing the fish tank

6. Interpretation of data

Materials

10 female Gambusia affinis, 5V Solar Power/DC Charging Oxygenator Fish Tank

Aquarium Air Pump Fishing Oxygen Pump Aerator, Medium size aquarium, improvise
rectangular water sprinkles, black printed plastic or 200 gsm paper, 5 meter cable wire,
5V electric supply, Mini V380 1080P IP Camera, 3 meter plastic pipes, synthetic
aquarium weeds
Risk and A. Fish tank
Safety Aquariums are a breeding ground for bacteria and, while many of those bacteria
are beneficial for the fish, there could be harmful bacteria present as well. To
protect the researcher from harmful bacteria, consider wearing rubber gloves when
cleaning the tank and will always wash the hand after coming into contact with tank
water. The risk for infection from contact with aquarium water is fairly low but if
open cuts or sores in hands, the risk of infection have may be elevated. Another
safety risk to consider is that, in rare cases, certain aquarium fish diseases can be
transmitted to humans. Tuberculosis, for example, is a deadly aquarium fish
disease that can be transmitted to humans from contact through open wounds or
sores with dead or infected fish. Another disease which can be transmitted to
humans is vibrio, a systemic infection which is typically transmitted through open
wounds. Though contracting disease from contact with aquarium fish is very rare, it
is still a risk of which should be aware when cultivating a freshwater tank. The
researchers will put sign that will keep students away from the fish tank

B. Power Supply
Another important aspect of aquarium safety is electrical safety. In order to keep
the fish tank running smoothly installing several pieces of equipment including filter
and lighting system. All of this equipment will need to be plugged into a wall outlet
and the proximity of that much electricity to all the water in the tank poses a very
real risk. One of the greatest risks in terms of electrical safety involves leaks and
drips – if water leaks or drips into the electrical supply, it could cause a short that
 might kill the fish in the tank or it could shock the person that touches the tank. To
prevent this from happening, researcher will ensure to arrange the cords of the fish
tank equipment. Prevent any water that drips down the cord from coming into
contact with the electrical supply. The researchers will put sign that will keep
students away from the fish tank

DATA ANALYSIS

The researcher will use pre observation and post observation. T-test will also be implied

BIBLIOGRAPHY
https://www.who.int/news-room/fact-sheets/detail/dengue-and-severe-dengue

Noreem M. et al. “Biological control of Mosquito larvae using edible fish.” Department of Veterinary
Parasitology, Sindh Agriculture University Tandojam-Pakistan.
https://www.researchgate.net/publication/324439144_BIOLOGICALCONTROL_OF_MOSQUITO_LARVAE_
USING_EDIBLE_FISH

Polverino et al. “Mosquitofish (Gambusia affinis) Preference and Behavioral Response to Animated Images
of Conspecifics Altered in Their Color, Aspect Ratio, and Swimming Depth.” Department of Mechanical and
Aerospace Engineering, Polytechnic Institute of New York University, Brooklyn, New York, United States of
America. http://sci-hub.tw/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3546983/

http://DOH.gov.ph