Research Plan

Project: Mortality and Aberrated Body Morphology of 3rd-early 4thinstar Aedesaegypti

Larvae exposed to the ethanolic leaf extract Bougainvillea spectabilis (Bogambilya).

Researchers: Joseph Samuel S. Athony (Team Leader)

Daniela M. De Chavez (Member)

Princess Joy S. Jarabe (Member)

Research Period: June 2018- September 2018

A. Rationale
Mosquitos are common especially the rainy season has already started. Excessive
water puddles because of heavy rainfall making the soil slow down when it comes to
absorbing the rain water causing an infestation of mother mosquitos to lay their eggs on
the water. Mosquitos lay their eggs in the water because that is where the larvae begin its
life. As we all know, mosquitos carry diseases that are undoubtedly lethal. The larvae of
mosquitos start of living in the water until they are ready to take off and start feasting like
their elders. Larvicides on the other hand were developed to limit the growth of larvae to
prevent them turning into harmful mosquitos. Larvicides are commonly made from plant
extracts that contain chemicals that harm the larvae which will result to the larvae dying
unable to continue its life. Thus, this study aims to determine if the Bogambilya leaf
extract is capable in eliminating the larvae limiting their chance into tuning into
mosquitos.

B. Questions or Problems Being Addressed

1. Is the extract from Bogambilya leaf effective to be a larvicide for Yellow fever
mosquito Larvae?
2. What is the morphology of the aberated body of the Aedesaegypti after the exposure
to the different treatments of the plant extracts?
3. What is the average mortality rate of the Yellow fever mosquito Larvae using
different treatments of Bogambilya leaf extract?
4. Is there a significant difference in the mortality rate of the Yellow fever mosquito
among the treatments?
5. What is the mode of action of the ethanolic extract of Bogambilya?

C. Goals/Expected Outcome/Hypothesis

Goal(1) To determine if the extract Bogambilya leaf is effective in killing the Yellow
fever mosquito (Aedesaegypti) Larvae
 Goal(2) To evaluate which treatment of Bogambilya leaf worked better than the others in
killing the Yellow fever mosquito (Aedesaegypti) Larvae
Expected Outcome: The extract of Bogambilya leaf could be an effective solution
against the Yellow fever mosquito (Aedesaegypti) Larvae.

Hypotheses:
Ho – There is no significant difference in the mortality rate among different
treatments ofBogambilya leaf on the Yellow fever mosquito Larvae.
Ha – There is a significant difference in the mortality rate among different
treatmentsofBogambilya leaf on the Yellow fever mosquito Larvae.

D. Description in detail of Methods or Procedures

D1. Procedures

A. Collection and Preparation of Plant Sample.

The Bogambilya plant will be gathered randomly within Bansud to Bongabong Oriental
Mindoro for identification in UP Institute of Biology in UP Diliman, Quezon City. After
the identification of the plant, the leaves are to be gathered. The leaves will be air dried
and crushed into to form of powder using a blender. After deconstructing the dried leaves
this will be transferred and kept in a clean and dry container. The plant will then undergo
phytochemical processes to determine the chemicals present within the plant.

B. Ethanol Extract Preparation

Five hundred fifty (500) grams of powdered Bogambilya leaves will be soaked and
moistened in two (2) liters of 95% ethanol for 48 hours. The ethanol will help extract the
desired component for the larvicide.

C. Extraction from Bogambilya Leaves

The Bogambilya leaves which were moistened will be prepared into the funnels with
cover for Maceration. The sample prepared will then undergo maceration overnight. The
macerated plant sample will undergo Rotary Evaporation followed by water bath.

D. Culturing Mosquito Larvae

The mosquito larvae will be cultured in an Ovicidal and Larvicidal trap, one for each
treatment. As the culturing proceeds, a total of One hundred fifty (150) larva need to be
used in the experimentation. The culturing process will be monitored until desired
amount of larvae is produced. 5 treatments are to be obtained each containing ten (10)
larvae for the treatment procedure to begin.
E. Application of Treatment
Five (5) treatments will be used to determine the effect of the plant extract towards the
larvae contained inside the OL (Ovicidal and Larvicidal) traps. The different treatments
will then be implemented within the OL traps. This process will be applied to all set ups
containing mosquito larvae. This process will be monitored after 24 hours after
implementing the treatment.

Treatment 1 = 100 mg/mL(Bogambilya leaf extract and distilled water)

Treatment 2 = 150 mg/mL(Bogambilya leaf extract and distilled water)
Treatment 3 = 200 mg/mL(Bogambilya leaf extract and distilled water)
Treatment 4 = crude bogambilya leaf extract
Treatment 5 = Negative Control(distilled water)

F. Experimental Design & Layout

An Ovicidal and Larvicidal traps with five treatments will be used. A total of50
Yellow Fever Mosquito larvae will be used in each treatment and will be observed after
24 hours. The number of deaths will be recorded afterwards after using the mortality rate
scale.

G. Statistical Analysis
Results will be expressed as means. To determine the significant difference in the
number of dead Yellow Fever Mosquito larvae among the treatments, One-Way Analysis
of Variance (ANOVA) will be used. For comparison of the treatment groups, Post Hoc
test (Scheffe) will be used with the significant threshold of 0.05.

H. Proper Waste Disposal

Disposal of waste will be highly observed throughout the study’s process. Chemical
wastes will be disposed according to their designated areas. Dead larvae will be out
casted after the experimentation. Some larvae that are still alive will be killed to prevent
the continuing of its growth. Other material used throughout the procedure will be
properly disposed following their standard procedures.

D2. Risk and Safety

Goggles, Laboratory gown, masks, and other Protective personal equipment (PPE) will be
used and observed during the extraction of the sample. Gloves and masks will also be used
during the collection and culturing of the mosquito larvae in the Ovicidal and Larvicidal
Traps.
D3. Data Analysis

Results will be interpreted as means. One-way Analysis of Variance (ANOVA) will be used
for comparison of the different groups of the treatment, Post Hoc test (Scheffe) will be used
with a significant threshold of 0.05.

E. Bibliography

Rajkumar, S., Jebanesan, A. (2007).Repellent activity of selected plant essential oils against
the malarial fever mosquito (Anopheles stephensi).Trop biomed, Volume 2, pp. 5-271.

Rahuman et al. (2000).Himalayan B. Effect of FeroniaIimonia on mosquito larvae

(Fitoterapia).Volume71, pp. 553-555.

Goswami, D., Rabha, B. (2007). Preliminary evaluation of mosquito larvicidal efficacy of

plant extracts. J Vector Borne Dis.). Volume 44, pp. 8-145.

Ghosh et al. (2006). Plant Extracts as Potential Mosquito larvicides.