LARVICIDAL POTENCY OF Arcangelisia flava (ABUTRA) STEM EXTRACT

AGAINST MOSQUITO LARVAE

In Partial Fulfillment of the Requirements in

PRACTICAL RESEARCH II:

Enriquez, Blessy

Ligutom, Princess Beatrice

Marwan, Al Dimar

Pecore, Krizel Babe

Serabani, Khizana Ameera D.

 TABLE OF CONTENTS

CHAPTER I.....................................................................................................................................
INTRODUCTION..........................................................................................................................1
1.1 Background of the Study......................................................................................................1
1.2 Aims of the Study.................................................................................................................3
1.3 Hypotheses...........................................................................................................................4
1.4 Significance of the Study......................................................................................................4
1.5 Scope and Delimitation.........................................................................................................5
CHAPTER II..................................................................................................................................6
REVIEW OF RELATED LITERATURE......................................................................................6
2.1 Conceptual Literature...........................................................................................................6
2.1.1 Mosquito Larvae ...........................................................................................................6
2.1.2 Mosquito Larvicide .......................................................................................................7
2.1.3 Arcangelisia flava (Abutra) ..........................................................................................8
2.2 Related Studies ....................................................................................................................9
CHAPTER III...............................................................................................................................14
METHODOLOGY.......................................................................................................................14
3.1 Research Locale..................................................................................................................14
3.2 Research Samples...............................................................................................................14
3.2.1 Arcangelisia flava (Abutra) ..........................................................................................8
3.2.2 Mosquito Larvae ...........................................................................................................8
3.3 Research Design.................................................................................................................16
3.4 Research Procedures...........................................................................................................16
3.4.1 Preparation of Samples ...............................................................................................16
3.4.2 Plant Extraction .............................................................................................................8
3.4.3 Preparation of Test Concentrations ...............................................................................8
3.5 Larvicidal Potential Bioassay..............................................................................................19
3.5.1 Mortality Rate ...............................................................................................................8
3.6 Data Collection ..................................................................................................................19
3.7 Statistical Analysis .............................................................................................................21
3.8 Waste Disposal ..................................................................................................................22
CHAPTER IV...............................................................................................................................23
RESULTS ....................................................................................................................................23
 4.1 Descriptive Statistics...........................................................................................................23
4.2 ANOVA .............................................................................................................................25
4.3 Post-hoc Analysis ..............................................................................................................27
CHAPTER V................................................................................................................................30
DISCUSSIONS............................................................................................................................30
CHAPTER VI...............................................................................................................................31
CONCLUSION AND RECOMMENDATIONS..........................................................................31
REFERENCES LIST ...................................................................................................................32
APPENDIX I................................................................................................................................35
RAW DATA.............................................................................................................................35
APPENDIX II...............................................................................................................................36
STATISTICAL ANALYSIS....................................................................................................36
APPENDIX II...............................................................................................................................37
RESEARCH MATERIALS......................................................................................................37
 ABSTRACT

Arcangelisia flava, commonly known as Abutra, is a plant native to Southeast Asia. It has

been traditionally used for medicinal purposes due to its pharmacological properties. The plant

contains various compounds like alkaloids, flavonoids, saponins, and terpenoids, which have

biological activities and defense against insects. One of its alkaloids, berberine, has shown

antibacterial properties and larvicidal activity. Mosquitoes are significant disease vectors,

causing millions of deaths annually. Plant extracts have been used as alternative mosquito

control methods for a long time. This study investigated the potential of A. flava stem extract as

a larvicide. This study utilized Randomized Complete Block Design (RCBD) with four (4)

treatments replicated 3 times. The following served as the treatments of the study. T1 – 3 grams,

T2 – 6 grams, T3 – 12 grams, and T4 – positive control (Commercial Larvicide). The results

showed that treatment and duration of exposure affected larval mortality. Treatment 4

(Commercial Larvicide) was found to be the most effective, with a mortality rate of 100% at all

time durations. Treatment 3 (12 grams) also had a high mortality rate across all time durations

and can be considered a viable alternative to Treatment 4. These findings emphasize the

importance of treatment type and exposure duration in reducing larval mortality. Further research

is needed to generalize these results to other populations or conditions. Overall, the study

provides valuable insights into A. flava extract as an alternative larvicide for mosquito control.
 INTRODUCTION
CHAPTER I

1.1 Background of the Study

Mosquitoes are the most important single group of insects in terms of public

health importance, which transmit several outrageous diseases like dengue, malaria,

filariasis, japanese encephalitis, etc. causing millions of deaths every year (Rathy et al.,

2015). Mosquito-borne diseases are transmitted to humans through the bite of infected

mosquitoes, which are considered major carriers of arboviruses (Souza-Neto et al., 2019).

Mosquitoes are highly mobile flying insects that can readily detect and avoid many

intervention measures. Therefore, targeting vector mosquitoes at the larval stage is the

best alternative since larvae are relatively immobile and confined within a given

geographical area, cannot change behavior to escape the effects of insecticides, and thus

become more vulnerable as compared to adult mosquitoes (Cheng et al., 2014).

Dengue is a large-scale health challenge in the Philippines (Undurraga et al.,

2017). From January 1 to December 17, 2022, the Philippines’ Department of Health

(DOH) reported 220,705 dengue cases, which is 182% higher compared to the 78,223

cases reported in the same period in 2021. While in Zamboanga City, the regional

Department of Health-9 has reported 3,884 dengue cases in 2022, an average of 971 cases

per quarter with 27 mortalities (Garcia, 2023).

Application of active toxic agents from plant extracts as an alternative mosquito

control strategy was available from ancient times (Ghosh et al., 2012). Plant extraction
can be important in the production of mosquito larvicides, as many plants contain natural

compounds that have larvicidal properties. These compounds can be extracted from the

plant and used to create effective larvicides that can be used to control mosquito

populations. Several studies have shown that various plants contain bioactive substances

with larvicidal activity, such as alkaloids, glycosides, phenolic compounds, flavonoids,

tannins, and saponins (Al-doaiss et al., 2021). The extraction of these compounds from

plants can be a cost-effective and environmentally friendly way to produce larvicides.

Additionally, using plant-derived compounds can reduce the risk of resistance developing

in mosquito populations, as they are less likely to develop resistance to natural

compounds compared to synthetic ones.

Arcangelisia flava, locally known as Abutra, is a plant species native to Southeast

Asia. This plant has been traditionally used for medicinal purposes due to its various

pharmacological properties. Arcangelisia flava contains secondary metabolites such as

alkaloid, flavonoid, saponin, and terpenoid compounds, all of which have the potential to

significantly produce biological activities and chemical defenses against insects (Chiet et

al., 2014). Arcangelisia flava contains berberine, an alkaloid which is present in the stem.

Berberine has been found to be active against gram-positive and gram-negative bacteria

(PROSEA, 2022) and has presented larvicidal activity greater than 95 % against

Rhipicephalus microplus larvae (Silva et al., 2021).

Despite the availability of various plant extracts containing bioactive compounds

with larvicidal activity, there is a lack of research on their effectiveness and feasibility as

a mosquito control strategy in the Philippines, particularly in Zamboanga City. The

 efficacy of these plant extracts in controlling mosquito populations, their environmental

impact, and their cost-effectiveness compared to conventional larvicides have not been

extensively studied in the context of the dengue outbreak in the region. This research gap

calls for further investigation and evaluation of the potential of plant-derived compounds

as a sustainable alternative to synthetic larvicides in controlling mosquito-borne diseases,

particularly in high-risk areas such as Zamboanga City.

1.2 Aims of the Study

This study aimed to determine the potential of Arcangelisia flava (Abutra) stem

extract as an alternative larvicide against mosquito larvae.

Specifically, this study sought to;

1. Determine which treatment with varying concentration of Arcangelisia

flava (abutra) stem extract has a lethal effect on mosquito larvae:

a. T1 – 3.0 grams of Arcangelisia flava (Abutra) Stem Extract + 100 mL

Distilled Water

b. T2 – 6.0 grams of Arcangelisia flava (Abutra) Stem Extract + 100 mL

Distilled Water

c. T3 – 12 grams of Arcangelisia flava (Abutra) Stem Extract + 100 mL

Distilled Water

d. T4 – 5 mL Commercial Larvicide + 100 mL Distilled Water

2. Evaluate the efficacy of the treatments in terms of:

 a. Mortality rate

b. Duration

1.3 Hypotheses
H0: there is no significant effect of using Arcangelisia flava (abutra) stem extract as

larvicide against mosquito larvae.

Ha: there is a significant effect of using Arcangelisia flava (abutra) stem extract as

larvicide against mosquito larvae.

1.4 Significance of the Study

Mosquitoes are persistent pests that spread some of humanity’s deadliest

diseases. The increase of the mosquitos’ population has become a main problem as it will

also increase the number of dengue carriers.

This research will benefit the community. It will greatly benefit everyone,

especially those who have been infected with mosquito-borne diseases. Since the

materials can be collected, it is possible to save money instead of buying expensive and

dangerous commercial pesticides.

This will also be beneficial for future researchers as they can use this study as a

guide for future research. They can also improve this study by using materials that are

more effective, exploit new ideas and use new methods.

 This study will provide information as regards on the effect of different amounts

of Arcangelisia flava (abutra) against mosquito larvae.

1.5 Scope and Delimitations

This study focused on the potential of Arcangelisia flava (abutra) stem extract as

an alternative larvicide against mosquito larvae. The raw materials were collected at

Katatagan, Brngy. Upper Calarian Zamboanga City. This study is only delimited in

determining the duration and mortality rate of the mosquito larvae after exposure to

different treatments, utilizing (RCBD). This study was conducted at Caldwell Adventist

Academy's Laboratory located at R.T. Lim, Boulevard Zamboanga City on April 10,

2023 to May 8, 2023.

 CHAPTER II
REVIEW OF RELATED LITERATURE
 

2.1 Conceptual Literature

2.1.1 Mosquito Larvae

The dengue vector Aedes aegypti (Linnaeus) belongs to the family Culicidae and

the order Diptera. Aedes aegypti is a known vector of several viruses including yellow

fever virus, dengue virus chikungunya virus and Zika virus (European Centre for Disease

Prevention and Control, 2023). The female A. aegypti preferably lay eggs in artificial

collections of water. The hatched larvae undergo growth and metamorphosis. In their life

cycle, four larval stages and the pupal stage are aquatic, and their adults are aerial.

Growth changes in form and size occur during their larval development (Bar & Andrew,

2013).

The morphology of A. aegypti larval body parts of head, neck, thorax, and abdomen

like mouth brush, palatium, mentum, compound eye, antenna, comb spines, siphon tube,

pecten teeth and anal papilla were described by various researchers. In 1 st instar stage, A.

aegypti larval head is narrow and triangular. In later stage, in the head capsule, convexity

appears. The head becomes large and attains a globular shape.

A. aegypti larvae are found in different aquatic habitats mainly in small water

collections. Various environmental factors like temperature, salinity, pH, dissolved

nutrients, and gases in the aquatic habitat influence the growth of mosquito larvae.
Extremes of temperature, lack of food and increased salinity result in reduced A. aegypti

larval growth and delayed development. (Bar & Andrew, 2013)

2.1.2 Mosquito Larvicide

Larvicides are a type of insecticide used to control mosquitoes indoors and

outdoors. They work by killing mosquito larvae and pupae before they can grow into biting

adults (Centers for Disease Control and Prevention, 2020). Larvicides target larvae in the

breeding habitat before they can mature into adult mosquitoes and disperse. Larvicide

treatment of breeding habitats helps reduce the adult mosquito population in nearby areas.

Liquid larvicide products are applied directly to water using backpack sprayers and truck or

aircraft-mounted sprayers. Tablet, pellet, granular, and briquet formulations of larvicides

are also applied by mosquito controllers to breeding areas (US EPA, 2023).

The major tool in mosquito control operation is the application of synthetic

insecticides such as organochlorine and organophosphate compounds. But this has not been

very successful due to human, technical, operational, ecological, and economic factors. In

recent years, use of many of the former synthetic insecticides in mosquito control programs

has been limited (Ghosh et al., 2012). Indeed, even if synthetic pesticides played a major

role in reducing the number of several diseases worldwide, their massive overuse has been

found responsible for resistance development in targeted vectors along with serious non-

target effects on human health and the environment (Ranson and Lissenden, 2016).

One of the most effective alternative approaches under the biological control

program is to explore the floral biodiversity and enter the field of using safer insecticides of
botanical origin as a simple and sustainable method of mosquito control. Further, unlike

conventional insecticides which are based on a single active ingredient, plant derived

insecticides comprise botanical blends of chemical compounds which act concertedly on

both behavioral and physiological processes. Thus, there is very little chance of pests

developing resistance to such substances. Identifying bio-insecticides that are efficient, as

well as being suitable and adaptive to ecological conditions, is imperative for continued

effective vector control management. Botanicals have widespread insecticidal properties

and will obviously work as a new weapon in the arsenal of synthetic insecticides and in

future may act as suitable alternative product to fight against mosquito borne diseases

(Ghosh et al., 2012). 

2.1.3 Arcangelisia flava (Abutra)

The genus Arcangelisia (A.), which belongs to the family Menispermaceae,

comprises 3 species (A. gusanlung, A. flava and A. tympanopoda) distributed in Southeast

Asia (Zhang et al., 1995; Suzuki et al., 2011). These plants possess various medicinal

properties and have wide-ranging applications in traditional and modern medicine.

Arcangelisia flava, locally known as Abutra, is a woody, perennial, climbing plant with a

very long stem growing from the ground level to the canopy of trees. It has been

traditionally used by local people for the treatment of several diseases, such as malaria,

dysentery, fever, abortion, the healing of hepatitis, indigestion, and as atonic agent. In

addition, A. flava was used as an important component of folk medicines for the treatment

of jaundice, smallpox, sore eyes, aphtha, water flea and as a stomachic and anti- helminthic

agent (Subeki et al., 2005).

 According to Maryani et al. (2013), phytochemical screening of A. flava yielded

alkaloid, flavonoid, saponin, and terpenoid compounds, all of which have the potential to

significantly produce biological activities and chemical defenses against insects (Chiet et

al., 2014). The plant yields several alkaloids: berberine, the principal alkaloid, with

jatrorhizine, columbamine and shobakunine. Several studies have shown that the ethanol

extract of A. flava stems has excellent antimicrobial activity against various microbes both

bacteria and fungi (Setyowati et al., 2014; Pratama, 2016). In a study conducted by

Solsoloy et al. (1987), crude aqueous extracts of A. flava showed insecticidal activity

against cotton bollworm (Helicoverpa armigera) in the Philippines. The pharmacological

effects of the plant are largely attributable to the alkaloid berberine, which is present in

concentrations of up to 5% in the stem (PROSEA, 2022).

Berberine is an alkaloid found in the barks, leaves, twigs, rhizomes, roots, and/or

antimicrobial, antiprotozoal, and antidiarrheal agent in Ayurvedic medicine and

positive as well as gram-negative bacteria, such as Diplococcus pneumoniae,

Escherichia hemolyticus and S. paradysenteria in different media. It had about the

same antibacterial against third instar larvae of A. aegypti (Paul et al., 2020). Berberine has

also presented in Philogène et al. (1984), larval, pupal, and adult survival of A. atropalpus

were significantly affected following treatment with berberine.

2.2 Related Studies

In the study ‘In vitro and in silico studies of the larvicidal and anticholinesterase

activities of berberine and piperine alkaloids on Rhipicephalus microplus’ conducted by Silva et

al. (2021), results showed that berberine has high in vitro larvicidal action on R. microplus. This
compound can be considered as a promising candidate for the development of new acaricidal

drugs.

In the study ‘Larvicidal Efficacy of Andrographis paniculata and Tinospora cordifolia

against Aedes aegypti: A Dengue Vector’ conducted by Paul et al. (2020), the study findings

suggest that the bioactive compound berberine can be a prospective candidate against 3 rd instar

larvae of Aedes aegypti.

 CHAPTER III
METHODOLOGY

3.1. Research Locale

 This study was conducted at Caldwell Adventist Academy Science Laboratory,

wherein the researchers extracted the A. flava (Abutra) stem varying in different

concentrations in determining the mortality rate and duration of the mosquito larvae.

3.2 Research Samples

3.2.1 Arcangelisia flava
A total of 500g (half-kilogram) of Arcangelisia flava (Abutra) were handpicked in

Katatagan, Brgy. Upper Calarian, Zamboanga City. The sample was then transferred to

Caldwell Adventist Academy Science Laboratory.

3.2.2 Mosquito Larvae

The 1st instar mosquito larvae were acquired by using a bucket filled with one liter

of tap water and was placed surrounded with plants. Once the 1 st instar larvae were

collected, it was moved to the Caldwell Adventist Academy Science Laboratory for

further preparation of test samples.     

 
3.3 Research design

            
Block 1 Block 2 Block 3 Block 4

T3 T1 T4 T3

T2 T4 T2 T1

T4 T3 T1 T2

Figure 1 – Experimental Layout in RCBD

             This study utilized Randomized Complete Block Design (RCBD) wherein 3

treatments with different concentration of A. Flava (abutra) stem extract were used with

randomly selected mosquito larvae. The study has 4 treatments: treatment 1 (3 grams A. flava

stem extract + 100 mL Distilled Water), treatment 2 (6 grams A. flava stem extract + 100 mL

Distilled Water), treatment 3 (12 grams A. flava stem extract + 100 mL Distilled Water),

treatment 4 positive control (Commercial Larvicide + 100 mL Distilled Water). Each

treatment has 3 replicates which totals to 12 replicates overall and in each replicate contain

fifteen (15) samples. A total of 180 random mosquito larvae were used.       

Legend:

T1 – 3 grams A. flava Stem Extract + 100 mL Distilled Water

T2 – 6 grams A. flava Stem Extract + 100 mL Distilled Water 

T3 – 12 grams A. flava Stem Extract + 100 mL Distilled Water

T4 – 5 mL Commercial larvicide + 100 mL Distilled Water

Sn - Sample Number

Figure 1 – Experimental Layout in RCBD

3.4. Research Procedure

3.4.1 Preparation of Sample

3.4.1.1 Arcangelisia Flava
The stems of Abutra were sorted and washed extensively to remove any

impurities or extraneous substances. Afterwards, they were dried and were cut into pieces

measuring about 2 inches long. These were then grounded using a grinder.

3.4.1.2 Mosquito Larvae

      After collecting the 1st instar larvae, the larvae were transferred to shallow buckets

containing 2 L of distilled water. Larvae were fed yeast at predetermined times and

observed daily. After 5 days, a homogenous population of late 3rd or early 4th instar

(3–5 days old, 4–5 mm long) was obtained. 

3.4.2. Plant Extraction

In a container, 180 grams of A. flava (abutra) stem each were placed. It was

immersed in 235.5 ml of 95 percent ethanol and sufficient distilled water to make a

415.5 ml solution. The A. flava (abutra) stem extract was stirred and stored in a cool,

dry area, covered, and carefully labeled. For 72 hours or 3 days, the container were
 stored in a cold, dry environment and applies maceration. After 3 days, the A. flava

(abutra) extract was filtered using filter paper to obtain the extracts. Purified extracts

were stored and labeled in a plastic container.

The filtered ethanolic extracts of A. flava (abutra) stem extract was evaporated

using open-dish extraction. It was accomplished by vaporizing water in a half-filled

beaker on an electric burner. The result of the open-dish evaporation of the A. flava

(abutra) stem extract would most likely be a residue that was correctly deposited in

another sterile vial. It was weighed digitally until a 72 grams residual of the extract is

obtained and stored in a vial appropriately labeled to prepare test concentration.

3.4.3 Preparation of Test Concentrations

                 Three (3) test concentrations were used and these are 3 grams A. flava (abutra)

Stem Extract + 100 mL Distilled Water, 6 grams A. flava (abutra) stem Extract + 100

mL Distilled Water, 12 grams A. flava (abutra) stem Extract + 100 mL Distilled

Water, Commercial Larvicide.  The treatment 1, a 3 grams of A. flava (abutra) stem

extract was dissolved in a 100 ml distilled water; for Treatment 2, a 6 grams of A.

flava (abutra) stem extract was dissolved in a 100 ml of distilled water; for Treatment

3, a 12grams of A. flava (abutra) stem extract was dissolved in a 100 ml of distilled

water; for treatment 4, a 5 mL Commercial Larvicide mixed with 100 mL Distilled

Water was used, it was utilized as the study’s positive control treatment. These

concentrations were utilized as the study’s experimental treatments. Each

concentration was added to a 100 ml of distilled water producing 3 different

concentrations from A. flava (abutra) and 1for commercial larvicide.  

 3.5 Larvicidal Potential Bioassay

             The study used twelve (12) containers, corresponding to the 4 treatments with 3

replicates. Fifteen (15) late 3rd or 4th instar mosquito larvae were placed in each of these

container using a dropper. Damaged larvae was removed and replaced.

      One hundred milliliters (100 mL) of each prepared concentration were applied to

15 mosquito larvae in the plastic container in three (3) replicates. It was remained on the

premises for 72 hours. 

3.6. Statistical Analysis

The data was analyzed using a statistical tool, and the mean, standard deviation, and

One-way analysis of variance (ANOVA) was employed at a 5% level of significance to

determine if there is a significant difference between the 4 treatments based on mortality

rate and time exposure.

Tukey’s Post hoc analysis was measured to determine which of the treatments is

significantly different based on mortality rate and time exposure.

 
 3.8. Waste disposal

 All waste should be properly disposed of. The area where the experiment was

carried out should be clean. This material, including its basin, should be disposed of

properly. Avoid spilled material and runoff dispersal, as well as contact with soil,

waterways, drains, and sewers.

 
 CHAPTER IV
RESULTS AND DISCUSSIONS

4.1 Descriptive Statistics

Table 1. Summary of 24-hour period mortality rate.

Treatment Mean Minimum Maximum Std. Std.Dev

Error

Treatment 1 0.2 0.133 0.333 0.0587 0.101

Treatment 2 0.17 0.066 0.266 0.0587 0.101

Treatment 3 0.53 0.466 0.6 0.0384 0.066

Treatment 4 0.77 0.666 0.86 0.0587 0.101

24 Hours
1

0.9

0.8

0.7

0.6

0.5

0.4

0.3

0.2

0.1

0
Treament 1 Treatment 2 Treatment 3 Treatment 4

Replicate 1 Replicate 2 Replicate 3

Figure
1. 24-hour mortality rate
 Based on the results provided, the column chart shows the mean mortality rates of

mosquito larvae treated with different treatments for a 24-hour duration.

The results suggest that treatments 3 and 4 were the most effective in terms of

mosquito larvae mortality rates. Treatment 3, which consisted of 12 grams of

Arcangelisia flava stem extract and 100 mL of distilled water, had a significantly higher

mean mortality rate compared to treatments 1 and 2, while treatment 4, which was the

commercial larvicide, had the highest mean mortality rate among all treatments.

Table 2. Summary of 48-hour period mortality rate.

Treatment Mean Minimum Maximum Std. Std.Dev

Error

Treatment 1 0.64 0.53 0.73 0.0587 0.101

Treatment 2 0.51 0.4 0.6 0..0587 0.101

Treatment 3 0.86 0.8 0.93 0.0384 0.066

Treatment 4 0.97 0.93 1 0.0222 0.038

Figure 2. 48-hour duration result

Based on the results provided, the mean mortality rates of mosquito larvae treated
with different treatments for a 48-hour duration are shown in the data.

Similar to the 24-hour duration, treatments 3 and 4 were the most effective in
terms of mosquito larvae mortality rates. Treatment 3, which consisted of 12 grams of
Arcangelisia flava stem extract and 100 mL of distilled water, had a higher mean
mortality rate compared to treatments 1 and 2, while treatment 4, which was the
commercial larvicide, had the highest mean mortality rate among all treatments.
 In addition, the mean mortality rates for all treatments were higher compared to
the 24-hour duration, which may suggest that a longer exposure time to the treatments
may lead to higher mortality rates of mosquito larvae.

Table 3. Summary of 72-hour period mortality rate.

Treatment Mean Minimum Maximum Std. Std.Dev

Error

Treatment 1 0.93 0.86 1 0.038 0.066

Treatment 2 0.75 0.66 0.86 0.058 0.101

Treatment 3 0.95 0.93 1 0.022 0.038

Treatment 4 1 1 1
72 Hours 0 0

1.2

0.8

0.6

0.4
Figure 3. 72-hour duration result
0.2

0
Treament 1 Treatment 2 Treatment 3 Treatment 4

Replicate 1 Replicate 2 Replicate 3

 Based on the data, it appears that all treatments had a high mortality rate after 48

and 72 hours. Treatment 4, the commercial larvicide, had the highest mortality rate at all

time durations, with a mean of 0.77 at 24 hours, 0.97 at 48 hours, and 1 at 72 hours.

Treatment 3, with 12 grams of Arcangelisia flava stem extract, had the second-highest

mortality rate at all time durations, with a mean of 0.53 at 24 hours, 0.86 at 48 hours, and

0.95 at 72 hours. Treatment 1, with 3 grams of Arcangelisia flava stem extract, had the

lowest mortality rate among the three extract treatments, with a mean of 0.2 at 24 hours,

0.64 at 48 hours, and 0.93 at 72 hours. Treatment 2, with 6 grams of Arcangelisia flava

stem extract, had a slightly higher mortality rate than Treatment 1 at all time durations,

with a mean of 0.17 at 24 hours, 0.51 at 48 hours, and 0.75 at 72 hours. Overall, it

appears that the alternative solution using Arcangelisia flava stem extract has potential as

a larvicide against mosquito larvae, with some treatments showing comparable efficacy

to the commercial larvicide.

4.2 ANOVA

Table 4. Summary of One-way ANOVA mortality-rate results for each time-duration.

Sum of Squares df Mean Square F Sig.

Between Groups 0.715185 3 0.238395 26.819 .000

24 Hours Within Groups 0.071111 8 0.008889

Total 0.786296 11
Between Groups 0.4011111 0.133704 20.055 .000
3
0.053333
48 Hours Within Groups 8 0.006667
3
Total 0.4544444 11
72 Hours Between Groups 0.103704 3 0.034568 8.484 .007
 Within Groups 0.032594 8 0.004074

Total 0.136296 11

The ANOVA results indicate that there is a statistically significant difference in the

mortality rates of mosquito larvae between the treatments at all time durations tested (24, 48, and

72 hours). This is supported by the low p-values (<0.05) for all three tests, indicating that the

probability of obtaining these results by chance is less than 5%. Additionally, the F-values for

each test are much larger than the F-critical value, further supporting the conclusion that the

differences between treatments are significant. Therefore, we can reject the null hypothesis and

conclude that there is a significant difference in the mortality rates of mosquito larvae between

the commercial larvicide and the alternative solution (Arcangelisia flava stem extract) at all

tested time durations.

4.3 Post-hoc Analysis

Table 5. Summary of Tukey HSD Post-hoc analysis mortality-rate results for the 72 hours
duration.
Comparison q-statistic p-value Significant
T1 vs T2 4.8242 0.0373 Yes
T1 vs T3 0.6030 0.9000 No
T1 vs T4 1.8091 0.5933 No
T2 vs T3 5.4272 0.0207 Yes
T2 vs T4 6.6332 0.0068 Yes
T3 vs T4 1.2060 0.8115 No
 The post-hoc analysis was conducted using the Tukey HSD test to compare the means of

each treatment group at the final duration of 72 hours. The results indicate that there were

significant differences in mortality rates between some of the treatment groups.

The results showed that there was a significant difference in mortality rates between

Treatment 1 and Treatment 2 (p = 0.0373239), Treatment 2 and Treatment 4 (p = 0.0067898),

and Treatment 2 and Treatment 3 (p = 0.0207396). This suggests that Treatment 2 had a

significantly lower mortality rate compared to Treatment 1 and significantly higher mortality

rates compared to Treatment 3 and Treatment 4.

On the other hand, there were no significant differences in mortality rates between

Treatment 1 and Treatment 3, Treatment 1 and Treatment 4, Treatment 3 and Treatment 4 (p >

0.05). This suggests that the mortality rates in these groups were not significantly different from

each other.

4.4 Discussions

Based on the results obtained from the experiment, it can be concluded that the mortality

rate of larvae was influenced by the treatment and the duration of exposure. The ANOVA results

showed that there were significant differences in mortality rates between the treatments at all

three time durations tested (24, 48, and 72 hours).

The post-hoc analysis revealed that Treatment 2 had a significantly lower mortality rate

than Treatment 1 and Treatment 3 at 72 hours, indicating that Treatment 2 was the most effective

in reducing the mortality rate of larvae in the experiment. Additionally, Treatment 4 had the

highest mortality rate, indicating that this treatment was the least effective.
 The findings of this study suggest that the type of treatment and the duration of exposure

are important factors that need to be considered when attempting to reduce the mortality rate of

larvae. It is also important to note that the results obtained in this experiment may not be

generalizable to other populations of larvae or to different experimental conditions.

 CHAPTER V
CONCLUSION AND RECOMMENDATIONS

5.1 Conclusion
Based on the results of the study, it can be concluded that the different treatments had

varying effects on the mortality rates of the larvae. Treatment 4 was found to be the most

effective, with a mortality rate of 100% at all time durations. Treatment 3 also had a high

mortality rate across all time durations and can be considered a viable alternative to Treatment 4.

Treatment 1 and Treatment 2 had lower mortality rates, with Treatment 2 showing the lowest

overall effect. The results of the ANOVA and post-hoc analysis further support these findings.

5.2 Recommendations
Based on the results of this study, the following recommendation is suggested:

1. The study recommends using Treatment 3 (12g A. flava stem extract + 100mL

Distilled Water) as an effective larvicide for mosquito larvae due to its high

mortality rate. Other researchers can explore different parts of A. flava with

bioactive compounds for potential larvicidal properties. Further research on

the plant is recommended.

2. Regular monitoring of mosquito populations and larval mortality rates should

be conducted to assess the effectiveness of larval control measures and to

identify any potential issues early on.

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 APPENDIX I

RAW DATA

Treatment Time Replicate Number of Number of Dead Mortality

Duration Larvae Larvae Rate
Treatment 1 24 1 15 3 0.2
Treatment 1 24 2 15 5 0.33333333
3
Treatment 1 24 3 15 2 0.13333333
3
Treatment 1 48 1 15 10 0.66666666
7
Treatment 1 48 2 15 11 0.73333333
3
Treatment 1 48 3 15 8 0.53333333
3
Treatment 1 72 1 15 14 0.93333333
3
Treatment 1 72 2 15 13 0.86666666
7
Treatment 1 72 3 15 15 1
Treatment 2 24 1 15 4 0.26666666
7
Treatment 2 24 2 15 3 0.2
Treatment 2 24 3 15 1 0.06666666
7
Treatment 2 48 1 15 9 0.6
Treatment 2 48 2 15 8 0.53333333
3
Treatment 2 48 3 15 6 0.4
Treatment 2 72 1 15 13 0.86666666
7
Treatment 2 72 2 15 11 0.73333333
3
Treatment 2 72 3 15 10 0.66666666
7
Treatment 3 24 1 15 9 0.6
Treatment 3 24 2 15 8 0.53333333
3
Treatment 3 24 3 15 7 0.46666666
7
Treatment 3 48 1 15 14 0.93333333
3
Treatment 3 48 2 15 13 0.86666666
 7
Treatment 3 48 3 15 12 0.8
Treatment 3 72 1 15 15 1
Treatment 3 72 2 15 14 0.93333333
3
Treatment 3 72 3 15 14 0.93333333
3
Treatment 4 24 1 15 12 0.8
Treatment 4 24 2 15 10 0.66666666
7
Treatment 4 24 3 15 13 0.86666666
7
Treatment 4 48 1 15 14 0.93333333
3
Treatment 4 48 2 15 15 1
Treatment 4 48 3 15 15 1
Treatment 4 72 1 15 15 1
Treatment 4 72 2 15 15 1
Treatment 4 72 3 15 15 1

APPENDIX II

RESEARCH SAMPLES

Arcangelisia flava
 APPENDIX III

PREPARATION OF SAMPLES

Washing, Drying, and Cutting of

Arcangelisia flava Stems

Acclimation of Mosquito Larvae

 APPENDIX IV

PLANT EXTRACTION

Immersing Arcangelisia flava with

Ethanol

Filtration after Maceration

Evaporation
 APPENDIX V

PREPARATION OF TEST CONCENTRATION

Arcangelisia flava Stem Extract

 APPENDIX VI

APPLICATION OF TREATMENTS

Testing the Arcangelisia flava Stem

Extract to the Mosquito Larvae

Data Collection