Dengue Mosquito borne

Zika Virus
Fever
diseases

• Japanese
• Malaria
Encephalitis

Chikungunya
Used coffee Caffeine
grounds content
LARVICIDAL
EFFECT OF USED
COFFEE
GROUNDS
AGAINST
MOSQUITO
LARVAE
• Highest incident rate each year, about 1 million people died of
the said disease (Oxitec, 2017)
• One of the deadliest animals in the world (Urdahl, 2017)
• The ability to carry and spread disease to the people not
infected with the disease (Urdahl, 2017)
• Usually, the female mosquitoes are the disease-carrying
mosquito because of the need of blood to nurture its babies
and able to reproduce more larvae.
• A lot of disease are
carried by
mosquitoes namely:
malaria, Zika virus
disease, dengue
fever and
chikungunya (CDC,
2016)
• Due to the certain
geographical area of
the Philippines, there
were mosquitoes
that can multiply
easily (DOST, 2008)
Dengue
• Dengue fever is a viral disease and is transmitted
by the bite female Aedes aegypti and
alborpictus.
• Aedes aegypti is the primary vector in spreading
the disease; it usually bites during the day.
• Dengue has become an all year-round disease
which become endemic in the country
(InterHealth Worldwide, 2016).
Chikungunya
• Chikungunya is a mosquito borne disease first
discovered in 1952 at Tanzania. It is an RNA virus
that belong to the genus Alphavirus and family
of Togaviridae.
• It is transmitted by the infected female
mosquitos or via contact with infected blood.
Malaria
• Malaria is also one of the most common mosquito-borne
disease in the world.
• It is considered a life-threatening disease and
transmitted through the bite of an infected female
Anopheles mosquito that carries
the Plasmodium parasite (World Health Organization,
2017).
Zika Virus
• Zika virus disease is a mosquito borne Flavivirus transmitted
by the Aedes mosquitoes, same mosquito that carries dengue,
chikungunya and yellow fever.
• Zika virus disease causes birth defects in babies whose mother
is infected with the disease. The infected babies are born with
undeveloped head and brain damage.
• It can be transmitted sexually, bite of a mosquito, blood
transfusion and can be found in blood, semen, urine, saliva
and the fluid in the eyes (WebMD, 2005).
• The most common symptom when you have Zika virus disease
is rashes, fever, joint pains, muscle pain and headache which
can last for days to a week.
Japanese Encephalitis
• it is a mosquito-borne flavivirus related to dengue, yellow
fever and West Nile viruses
• Permanent neurologic or psychiatric sequelae can occur in
30%–50% of those with encephalitis.
• There is no cure for the disease & the treatment is focused on
relieving severe clinical signs and supporting the patient to
overcome the infection. (WHO, Dec 2015)
Mosquitoes
• are of major medically importance due to their role as vector
for transmission of various diseases.
• Mosquitoes from the subfamilies Anophelinae and Culicinae
are public health important that provide a piercing-sucking
mouth parts several times longer than the head.
Morphology
• They are Holometabola with the first stage differing
completely from the last one in form, structure and habits.
• Most of the common and important mosquitoes as
transmitter of pathogens for animal and man belong to four
genera: Anopheles, Culex, Mansonia and Aedes. (Companion
Vector Borne Diseases, 2017).
Development
• The adult life span can range from two weeks to a month
depending on environmental conditions.
• The life cycle can be completed one and a half to three weeks.
• The larval stage is completed within 4 to 9 days
• Adult: Male and female adults feed on nectar of plants;
however, female mosquitoes need blood to produce eggs, and
are active in the daytime (Center for Disease Control and
Prevention, 2012).

Habitat
• Mosquitoes prefer stagnant water within which to lay their
eggs. They most commonly infest ponds, marshes, swamps
and other wetland habitats.
Feeding Habits
• They need protein found in blood to develop their eggs.
• They choose their targets through a combination of smell,
heat and visual cues, and continue seeking blood meals until
their abdomens are full.
• They feed every two or three nights during that time.
• Using sensors on their antennae to detect trace amounts of
carbon dioxide released in human breath.
• They feed on a lower frequency of other hosts including
bovine, swine, cat, rat, and chicken.
 Mosquito Ovicidal-Larvicial
(OL) Trap
• It is made to address the problem
in dengue infection in the
Philippines by inhibiting its early
stages.
Coffee
• high content of compounds that exert antioxidant and
other beneficial biological properties.

Properties
• they are stronger, more resistant to pests and disease
• higher temperatures: 24-30°C (75-86° F)
• rainfall: 2000–3000 millimeters (79-118 inches)
• grows in lower elevations
• sea level to 700 meters (about 2,300 feet)
• 40-50 % higher caffeine levels
Caffeine (1,3,7-trimethyl-xanthine)
• Classified as a central nervous stimulant and
an ergogenic compound.
• Metabolized primarily in the liver, with
assistance from the kidneys and brain.
• Stimulates the central nervous system as an
adenosine-receptor antagonist.
• Natural component of tea, guarana,
chocolate and coffee.
Effects of Caffeine to
mosquitoes
• Strengthens the validity of considering used coffee
ground preparations as possible alternative control of
Aedes aegypti. (Guirado and Bicudo, 2007)
• Attractiveness of bio insecticides caffeine and used
coffee grounds in the choice of oviposition site by Aedes
aegypti (Guirado and Bicudo, 2015)
Theoretical Framework
• Previous studies have been discovered that the used of coffee
grounds help to reduce the rapid growth population of
mosquito particularly Aedes aegypti. Coffee contains large
amounts of chemicals many of which are high toxic to the
developing larvae of mosquito which has been hypothesized
by Satho et al. (2015). Coffee also decreases the reproductive
capacity of mosquitoes which helps to lessen the laying of
eggs for female mosquitoes (Guirado & Bicudo, 2015).
Considering the importance of the oviposition site in the
reproductive success, the caffeine will interfere in the choice
of the oviposition site by Aedes aegypti(Guirado & Bicudo,
2015). With the following literatures, it was suggested that
caffeine can be used for alternative insect control. The afore-
mentioned studies played an important role in developing our
research study.
Conceptual Framework
Independent Variable Dependent Variable
Coffee ground Mortality rate of
concentrations (5 days mosquito larvae
incubation)
• 0.5 g/ml
• 1.0 g/ml
• 1.5 g/ml
• 2.0 g/ml
• 2.5 g/ml
• 3.0 g/ml
Positive Control
• Mosquito OL Trap
Negative Control
• Distilled water
Statement of the Problem
This study aims to investigate the larvicidal effect of used coffee
grounds against mosquito larvae and specifically answers the
following:
1.) What is the mortality rate of mosquito larvae when expose
to different concentrations used coffee grounds:
1.1.) 0.5 g/ml;
1.2.) 1.0 g/ml;
1.3.) 1.5 g/ml;
1.4.) 2.0 g/ml;
1.5.) 2.5 g/ml and;
1.6.) 3.0 g/ml?
Statement of the Problem
2.) What is the mean mortality rate of mosquito larvae at 5
days incubation in Mosquito OL trap?
3.) Is there a significant difference in the mean mortality rate
of the different concentration of coffee and the Mosquito OL
trap?
Null Hypotheses
• Ho1: Hypothesis free
• Ho2: Hypothesis free
• Ho3: There is no significant difference in the mean mortality
rate of mosquito larvae when exposed to different volumes of
the coffee grounds and in Mosquito OL trap.
CHAPTER II
METHODS
Design
• Study will be a experimental post test with control
design
• Allocated to experimental group and control group
• In our experiment:
① Coffee concentration (experimental group)
② Positive and negative control (control group)
③ Mortality rate of mosquito larvae
Setting
• Mindanao Medical School Foundation College
laboratory
Subjects
• Used coffee grounds
• mosquito larvae
• Mosquito OL Trap
Samples/Specimens
• Used coffee grounds
• ordered from selected coffee shops in Davao City
• Mosquito larvae
• isolated in Sarphil Village, Agdao, Davao City
• pass through four instars
• fourth-instar larva was longer than the other (8 mm long)
• based on time and sizes
• Mosquito OL trap
• requested from DOST
Measures
• To determine the significant difference in the
mortality rate of mosquito larvae in different
concentrations of aqueous used coffee grounds
suspensions
One way ANOVA
• used when comparing two or more group
means on a continuous dependent variable.
• standardized mortality ratio
• calculated by dividing the observed number of
dead cases by the expected number of death
 Procedures
A. Collection of used coffee grounds
The used coffee grounds will be ordered from selected coffee
shops in Davao city and will be stored up until experimentation in
Mindanao Medical Foundation College.
B. Preparation and screening of treatments
The different concentrations of used coffee grounds are mixed
with 200 mL of water to make an aqueous used coffee ground
suspension. The procedures are as follows:
b.1. Label 13 black cups with the following: 2 cups for 0.5g/ml, 2
cups for 1.0 g/ml, 2 cups for 1.5 g/ml, 2 cups for 2.0 g/ml, 2 cups for
2.5 g/ml, and 2 cups for 3.0 g/ml for coffee concentrations, 2 cups for
OL Trap as positive control, and 1 cup for distilled water for negative
control.
Procedures
b.2. Place the 250 ml of different concentrations of coffee
extract in the different black cups.
b.3. In OL Trap, place 250 ml of water then add 2 packs of
pellets in its cups.
b.4. Place ten (10) larvae in each black cup.
b.5. Cover with fine mesh fabric fixed with rubber band.
b.6. Three (3) batches will be conducted.
b.7. Check the set-up daily, same time of the day.
b.8. After 5 days incubation, check for live and dead larvae in
different set-ups of concentrations and record.
 Approval of the proposal

Transmittal Letters will be sent to the Department Dean and Director of Laboratories

Implementation of the Experiment

Isolation of Mosquitoes

Breeding of Mosquitoes

Collection of Used Coffee Grounds

Preparation of Treatments and Controls. 0.5 g/ml, 1.0 g/ml, 1.5 g/ml, 2.0 g/ml, 2.5 g/ml, 3.0 g/ml
for coffee concentrations, Mosquito OL Trap (2 packs of pellets dissolved in 250 ml water) and
distilled water for positive and negative control

Larvae will be transferred accordingly in each cups that contained different volumes of used coffee
grounds, positive, and negative control.

Data will be collected after five (5) days of incubation

Experiment will be carried out for three (3) trials to ensure certainty of results.
Methods
• Guirado and Bicudo (2007)
Suspension of caffeine
Alternative control of Aedes aegypti
Target stage: 3rd instar larvae
Biosafety considerations
• Many factors are needed to be considered when dealing
biological organisms because it is not only hazardous but
also takes time.
• Wearing of proper personal equipment is a must. When
dealing with mosquitoes, your skin needs to be fully
covered to avoid the possibility of acquiring mosquito-
borne diseases.
• Proper disposal of chemicals should be observed to avoid
spreading harm to another incognizant individual
following the seminar that will be conducted at the
Department of Health Region XI.
Biosafety Considerations
• The individuals performing the experiment should be
knowledgeable of the proper handling of specimen to
avoid unnecessary accidents. It should be noted when
lives of any organism, in our case, the lives of the
mosquito should be considered. The mosquitoes should
be handled in an enclosed area where they could not
escape and may potentially spread an endemic.