Morphological Deformities and Abnormal Behavioral Responses of Fourth Instar Aedes

aegypti (Mosquito) Larvae Subjected to Multifarious Residential Commodities

A Science Investigatory Project

Submitted by:

Krizzia Eve Grace P. Alzate

Hillaenna Heart Delmo

Justine Rose Oropesa

Avril Mampusti

Elaine Villanueva

King Lara

Ken Gabriel Fajarito

John Nicole Quinico

Submitted to:

Romnick Ureta

BANSUD NATIONAL HIGH SCHOOL

Regional Science High School for Region IV-B (MIMAROPA)

Pag-asa, Bansud, Oriental Mindoro

 INTRODUCTION

The predominance of insect vectors such as mosquitoes in tropical and subtropical

countries has resulted to the transmission of serious human, animal and plant diseases that causes

millions of death annually. Mosquito is the most indisputable medicinal significant arthropod

vector of diseases. The vector-borne diseases caused by mosquito are one of the major health

problems in most of the countries. It is affecting the socio-economical status of many nations.

They transmit parasites and pathogens which continue to have disadvantageous impact on human

beings The diseases like filariasis, dengue, yellow fever, malaria, Japanese encephalitis, and

chikungunya are some of the deadly diseases spread by mosquitoes. And for tropical countries

like the Philippines, dengue is one of the most prevalent.

Dengue is vector-borne disease with 390 million infections per year extending over 120

countries of the world. According to the Department of Health (DOH), 622 people have died due

to dengue as of July 20 this year. These deaths came from the 146,062 dengue cases from

January to July – a number 98% higher than the recorded incidence during the same period last

year. Aedes aegypti is a primary vector for dengue viral infections for humans. This has

necessitated the need for research and development of environmentally safe, biodegradable, and

low cost indigenous method for vector control, which can be used with minimum care by

individuals and communities.

Current focus on application of natural product against mosquito vectors has been the

main priority for research due to its eco-safety. The extensive use of chemical insecticides has

led to severe health problems, environmental pollution, toxic hazards to human and non-target

species, and development of insecticide resistance on mosquitoes. Hence, there is a need for
developing biologically active natural chemical constituents which act as a larvicidal and

promising to reduce the risk to humans and harmful accumulated residues.

This study will be focused on as a biological control agent against Ae. aegypti larvae. To

date, no other studies have been conducted with residential commodities such as Malus pumilla

mill (Apple cider vinegar), Allium sativum (Garlic) juice, Sodium hypochlorite (Bleach), Saline

water (Saltwater) and Hydrochloric acid (Muriatic acid) for its larvicidal efficacies and

morphological and behavioural responses on Ae. aegypti larvae.

METHODOLOGY
Collecting of Aedes aegypti larvae

Ae. aegypti larvae were collected from Brgy. Poblacion, Bansud, Oriental Mindoro. All

replications were transported with 500 mL of water to the laboratory and kept at a room

temperature for 24 hours. The samples were identified as fourth instar larvae using a digital

microscope.

Preparation of Multifarious Residential Commodities

Malus pumila mill

20% of Malus pumila mill was dissolved in 80% of distilled water. From the solution,

different set-ups were serially observed for 1, 24, and 72 hours.

Saline water

20% of Saline water was dissolved in 80% of distilled water. From the solution, different

set-ups were serially observed for 1, 24, and 72 hours.

Sodium hypochlorite

20% of Sodium hypochlorite was dissolved in 80% of distilled water. From the solution,

different set-ups were serially observed for 1, 24, and 72 hours.

Allium sativum juice

20% of Allium sativum juice was dissolved in 80% of distilled water. From the solution,

different set-ups were serially observed for 1, 24, and 72 hours.

Hydrochloric acid
20% of Hydrochloric acid was dissolved in 80% of distilled water. From the solution,

different set-ups were serially observed for 1, 24, and 72 hours.

Morphological View and Visualization

Ae. aegypti fourth instar larvae was observed under the electronic digital microscope,

magnification of 0-1000X.

Record of deformities
During the course of lethal experiments, the morphological features of Ae. aegypti fourth

instar larvae from treated and controlled set-ups were compared. Any notable difference in

appearance between the treated and controlled set-up was recorded as deformity.
 RESULTS AND DISCUSSION

1a 1b

1c 1d

Figure 1. Behavior of Ae. aegypti fourth instar larvae exposed to Malus pumila mill

Figure 1b. Behavior of Ae. aegypti fourth instar larvae exposed to Malus pumila mill for 1 hour.

By exposing the larvae to the solution of 20%Malus pumila mill and 80% distilled water and

observing it for 1 hour, it was determined to staying still and shows no existence of life. There

are no visible changes in the structure of the larvae.

Figure 1c Behavior of Ae. aegypti fourth instar larvae exposed to Malus pumila mill for 24 hours.

As observed it was determined that the larvae was killed. The visible is its body becoming thin

and became longer. It also shows brown pupa with some melanization.

Figure 1d Behavior of Ae. aegypti fourth instar larvae exposed to Malus pumila mill for 72 hours.
As observed, it became a whitish color larvae with an opaque swelling thorax. It was determined

to be dead.

2b

2a

2c 2d
Figure 2. Behavior of Ae. aegypti fourth instar larvae exposed to Saline water

Figure 2b. Behavior of Ae. aegypti fourth instar larvae exposed to Saline water for 1 hour. By

exposing the larvae to Saline water and observing it for 1 hour, it was showing a normal

behavior for a larvae. No visible changes in the structure of the larvae are checked.

Figure 2c. Behavior of Ae. aegypti fourth instar larvae exposed to Saline water for 24 hours. The

body changes its length and have some deformities.

Figure 2d. Behavior of Ae. aegypti fourth instar larvae exposed to Saline water for 72 hours. It

exhibits dwarf pupa retarded abdomen.

 3a

3b

3c

Figure 3. Behavior of Ae. aegypti fourth instar larvae exposed to Sodium hypochlorite

Figure 3b. Behavior of Ae. aegypti fourth instar larvae exposed to Sodium hypochlorite for 1

hour. As observed it was determined that the larvae was killed. It started losing color which

turned into white.

Figure 3c. Behavior of Ae. aegypti fourth instar larvae exposed to Sodium hypochlorite for 24

hour. As observed it was determined that the larvae was killed. The visible changes of the larvae

is the color; the larvae became visible white.

Leaving it for 72 hours, the body had dissolve into the concentration.
 5a

5b
5d

5c
Figure 5. Behavior of Ae. aegypti fourth instar larvae exposed to Hydrochloric acid
Figure 5b. Behavior of Ae. aegypti fourth instar larvae exposed to Hydrochloric acid for 1 hour.

As exposing it to the solution of muriatic acid and water, it started showing a behavior in which

its wiggling too much and stayed still after that. no visible changes of the body is determined.

Figure 5c. Behavior of Ae. aegypti fourth instar larvae exposed to Hydrochloric acid for 24

hours. It was determined that the larvae is lifeless and the changes to its body is that it loss the

black color in the abdomen which became white and the head became red and also the thorax

swelled.

Figure 5d. Behavior of Ae. aegypti fourth instar larvae exposed to Hydrochloric acid for 72

hours. It was determined that the larvae is lifeless and the changes to its body is that it became

white and the head became red and also the whole body swelled.
 6a

6b

6c
Figure 6. Behavior of Ae. aegypti fourth instar larvae exposed to Distilled water as

Controlled Set-up

Figure 6b. Behavior of fourth star larvae exposed to Distilled water for 1 hour

By exposing it to distilled water, it still acts normal. No visible changes are checked.

Figure 6c. Behavior of fourth star larvae exposed to Distilled water for 24 hours

As observed, it evolves into a pupa. The body structure change into a pupa

Figure 6d. Behavior of fourth star larvae exposed to Distilled water for 72 hours

Evolved into mosquito.

 CONCLUSION

In conclusion, discovering an environmentally friendly insecticide to control mosquito

vectors is considered to be a vital role which reduces the negative impacts caused by chemical

insecticides in our environment. Thus, this current research provides evidential framework for

malus pumilla mill (apple cider), allium sativum (garlic) juice, sodium hypochlorite (bleach,

saline water and hydrochloric acid(muriatic acid) as potential larvicidal agents. (Kuha ng onti sa

results ) On the basis of the above results, we can conclude that these residential commodities

has a paramount larvicidal importance. The synthetic products used can be obtained easily at a

very low cost. The result clearly reveals that these domestic items could serve as a potential

larvicidal agents against the dengue vector A. aegypt Furthermore, its application may alter the

morphology and behavioural process of Ae. aegypti larvae. Therefore, it can be concluded that

the said residential commodities have desired compounds in larvicidal bioassays and

substantially reduce the possibilities of physiological resistance development in mosquito

population.