Butterfly Pea (Clitoria ternatea) flower extract as an Alternative

Solution for Ant Control

ANSALE, SHEENA MARIE U.

JAGONIA, ALYSSA JEANNE S.
JENKINS, JADEN ART HOWARD G.
LABADAN, TONI SHEKAYENA E.
NUENAY, FAYE V.
SERONAY, DENYEL ONA BRIANNA
TAMPOS, JHULIA ALEXIS B.

A Research Proposal
Submitted to the Faculty of
Integrated Developmental School
Mindanao State University at Naawan
9023 Naawan, Misamis Oriental
In Partial Fulfillment of the
Subject

Research 2

February 2025
 INTRODUCTION

1.1. Background of the study

The infestation of red fire ants (Solenopsis invicta) has become a problem for
households, shops, and other establishments. These insects are spreading quickly, causing
infrastructure damage and caused by their nests for both homeowners and shopkeepers. This
issue has grown into a concern, as many places are experiencing considerable harm and
inconvenience.

Clitoria ternatea, also known as butterfly pea, might be a great solution. This plant,
with its bright blue flowers, has been studied for its insecticidal properties on other insects.
This research aims to determine how effective butterfly pea flower extract is at controlling
the population red fire ants (Solenopsis invicta) and whether it can be a good, eco-friendly
alternative to ant control. By exploring this natural option, the researchers hope to find a safer
and more environmental way to control red fire ants (Solenopsis invicta).

The aim is to harness the strength of butterfly pea flower extract as an alternative for
ant control, offering a natural and efficient substitute for conventional synthetic’s. By using
this extract, it can reduce the detrimental effects of the Red fire ants more sustainably,
without harming humans, animals, and the environment. Additionally, by researching
potential novel and natural ant control, we can help advance the development of agriculture
practices that are more environmentally friendly and contribute to a healthier, greener future.

Additionally, it focuses on a harmful species of ant that can cause destruction and
economic damage if left unchecked. It is also important because of the growing awareness of
the need to protect and preserve our environment, making research into green and sustainable
pest control methods an intriguing topic.

In agriculture, insect pests damage crops, leading to reduced yields and economic
losses for farmers. Public health is also at risk, as many insects spread diseases like malaria,
dengue, and Zika virus. Control in insect these disease-carrying insects and protect public
health. Economically, pests cause significant losses in agriculture, forestry, food storage, and
manufacturing. Effective help reduce these losses. Developing new, eco-friendly pests
control is essential to maintain ecological balance and reduce these issues
1.2. Statement of the problem
1. How effective is Clitoria ternatea extract in repelling or eliminating ants compared to
commercially available ants control solutions?
2. What is the optimal concentration of Clitoria ternatea extract for maximum
effectiveness in ant control?
3. How does the application of Clitoria ternatea extract affect the behavior and mortality
rate of ants over a specific period?

1.3 Null Hypothesis

There is no significant difference between the efficacy of the different concentrations
of Butterfly Pea (Clitoria ternate) flower extract as an insecticide.

1.4 Significance of the study

The study focuses on analyzing the effectiveness of flower extract from the Butterfly Pea
plant as an alternative for ant control against Red fire ants and thus, it exhibits a unique
viewpoint by presenting a new, environmentally friendly method of pest management. This
research not only has the potential to reduce environmental harm but also introduces a fresh
and effective approach to managing pests sustainably. Specifically targeting pests like ants,
can revolutionize how to control these nuisances, offering more environmentally-friendly
solutions. In addition to having the potential to reduce environmental impact, this research
represents a novel contribution to sustainable pest management and can reverse established
agricultural and biodiversity preservation paradigms.

1.5 Scope and limitation

This study focuses on the butterfly pea flower extract as an alternative for ant control,
in preventing the Red Fire Ants from entering the inhabitant’s residence. The researchers
will be utilizing only the full-blown flower excluding the buds of that species to create
efficient and an eco-friendly way to control these invasive ant species (Selonopsis
Invicta) so the researchers will be able to investigate the Butterfly Pea (Clitoria ternate)
flower extract as an alternative in population control.
1.6 Conceptual Framework
The conceptual framework of the study depicts all of the independent and dependent
variables relevant to our study. The independent variables, which are the butterfly pea
(Clitoria ternatea) flower extract and red ants (Solenopsis invicta), will be evaluated
using the dependent variables provided. This study proposes how varying concentrations
of Butterfly Pea (Clitoria ternatea) flower extract and Red Ants (Solenopsis invicta) will
likely influence various dependent variables. Using a conceptual framework, this study
will examine the independent variables and assess their effects on dependent variables.

Independent Variable Dependent Variable

Butterfly pea flower Red ant

extract Ant Control

Figure 1. Conceptual framework of the study

1.7 Definition of terms

To facilitate the understanding of this study, different terms are defined herein.
Ant Control. refers to the methods and strategies used to manage or eliminate
ant infestations in homes, agriculture, and other environments.

Infestation. refers to the presence and rapid multiplication of unwanted

organisms, such as insects, rodents, or parasites, in a particular area, causing harm or
nuisance.

Infrastructure. refers to the fundamental physical and organizational

structures, facilities, and systems needed for the operation of a society, economy, or
institution, such as transportation, communication, power supply, and water systems.

Inconvenience. refers to a state of discomfort, difficulty, or disruption that

affects normal activities or causes annoyance and inefficiency.
 Alternative. refers to an option or choice that serves as a substitute for
something else, often considered as a different approach or solution to a problem.

Harness. refers to a device or system designed to control, secure, or utilize the

power or strength of something.

Conventional. refers to something that is based on established customs,

practices, or standards.

Synthetic. refers to something that is artificially created or manufactured,

typically by combining various elements or compounds, as opposed to being naturally
occurring.

Detrimental. refers to something that causes harm, damage, or negative effects.

It suggests that an action, substance, or situation has a harmful impact on a person,
system, or environment.

Malaria. is a serious infectious disease caused by Plasmodium parasites,

transmitted through the bite of infected female Anopheles mosquitoes.

Dengue. is a viral illness transmitted by Aedes mosquitoes, particularly. It is

characterized by sudden high fever, severe headaches, pain behind the eyes, joint and
muscle pain, rash, and nausea. In severe cases

Zika virus. is an infection caused by the Zika virus, confirmed through blood
or urine tests. The impact is measured by the incidence of infections, the occurrence
of birth defects, and public health response efforts in affected areas.

Intriguing. refers to something that arouses curiosity, interest, or fascination

due to its unusual, mysterious, or thought-provoking nature.
 Viewpoint. is a particular perspective, opinion, or way of understanding and
interpreting a subject, event, or issue.

Nuisances. refer to anything that causes annoyance, inconvenience, or harm,

disrupting comfort, health, or well-being.

Revolutionizing. means to bring about a radical or fundamental change in a

system, process, or field, often leading to significant improvements or advancements.

Biodiversity. refers to the variety of life forms within a given ecosystem,

including different species of plants, animals, fungi, and microorganisms, as well as
the genetic diversity within species and the diversity of ecosystems.

Depict. means to represent, describe, or illustrate something visually, verbally,

or symbolically.

Concentrations. refer to the amount or proportion of a substance within a

specific volume or area.

Extract. means to remove, obtain, or separate something from a larger whole,

often through a specific process.

Excluding. refers to the act of leaving out, omitting, or preventing someone or

something from being part of a group, category, or process.

Invasive. refers to something that spreads aggressively or intrusively, often

causing harm to the environment, ecosystem, or system it enters.

Utilizing. refers to the act of making use of something effectively or efficiently

to achieve a particular purpose or goal.
 Exhibits. refer to objects, displays, or presentations that are shown to the
public or an audience, often for educational, informative, or artistic purposes.

Substitute. refers to an item, person, or method that replaces another, typically

serving the same function or purpose.

Efficient. generally, refers to the ability to achieve a desired result with the
least number of resources.

Sustainability. refers to the ability to maintain or support a process or system

over the long term without depleting resources or causing harm to the environment,
economy, or society.

Agriculture. is the practice of cultivating soil, growing crops, and raising

animals for food, and other products used to enhance human life.

Yields. in the context of agriculture refer to the amount of crop or agricultural

product that is produced per unit of land area, often expressed in terms of weight, and
volume.

Exhibit. something means to show or display it in a visible or public way,

often to demonstrate or prove a point.
 2. REVIEW OF RELATED LITERATURE AND STUDIES

2.1 Butterfly Pea Plant

‘Butterfly Pea (Clitoria ternate) also known as the Asian Pigeon Wings, Blue Bell
Vine, Blue Pea, Cordofan Pea, and Darwin pea, is a marvelous herb that originates from
tropical Asia, more precisely from the equatorial region.

Derived from a plant that is common to most Southeast Asian countries. Blue ternate
(Clitoria ternate). is a potential source of phytochemicals and hormones with nutritional and
helpful benefits. The plant exhibits morphological variations, particularly in flower color and
structure, which contributes to its genetic richness, according to (Suarna,2021).

Butterfly pea (Clitoria ternate) is a multi-purpose forage legume. It is adaptable to a

wide range of temperatures, rainfall, and altitude Butterfly pea, a plant that has gathered
attention on account of its potential as an eco-friendly pesticide, is the origin of Sero-X.
Butterfly pea (Clitoria ternatea) stands out as the sole leguminous plant known for its
production of stable cyclic plant defense peptides called cyclotides. These cyclotides have
garnered significant attention for their potential as eco-friendly pesticides, exemplified by the
recent registration of a butterfly pea extract, Sero-X®, for agricultural use.

The study focuses on exploring the variability in cyclotide expression and insecticidal
properties among butterfly pea accessions sourced globally. Analyzing peptide extracts from
23 accessions across 11 countries, we identified substantial diversity in cyclotide expression.
Some accessions lacked the typically abundant cyclotide Cter M, attributed to missense
mutations in CterM-like precursor genes. Additionally, one accession showed undetectable
levels of other cyclotides (cliotide T1, cliotide T4, Cter A, and Cter Q). Surprisingly,
cytotoxicity against Sf9 (Spodoptera frugiperda) cells was not solely dependent on Cter M, as
accessions lacking this peptide still exhibited cytotoxic effects. These findings contribute
fundamental insights for the selective breeding of butterfly pea varieties with enhanced
insecticidal properties. (Gilding et al., 2019)

Pesticide usage has undeniably played a significant role in enhancing agricultural

productivity, both in terms of yield and quality, thereby boosting farmers' incomes, especially
in developed nations. However, improper application of pesticides—without following safety
guidelines—has resulted in serious health hazards for humans, wildlife, and the environment.
This includes risks from exposure to farm workers, contamination of air and water, and the
presence of pesticide residues in food products. Consequently, there has been an increasing
demand for food safety and quality over recent decades, leading to stringent safety
regulations on imports and strict limits on pesticide residues in commodities. Additionally,
higher standards for product quality continue to be established.Public awareness regarding
the harmful impacts of pesticides on food safety and environmental health has risen
significantly in recent years. This has intensified the search for alternatives to conventional
chemical pesticides, such as biopesticides. As a result, the pesticide industry has experienced
substantial transformations over the past few decades. These changes have led to more
efficient pesticide use through advancements in pest management technologies and practices
within Integrated Pest Management (IPM) frameworks. Such developments have improved
pest management strategies, reduced pesticide consumption in some instances, and slowed
the increase in demand for chemical pesticides.

2.1.1. Chemical Properties of Butterfly Pea

The butterfly pea plant, a member of the pea family, is generally inedible by humans
and contains toxins in its root and seeds. Eating causes serious digestive upset, including
nausea, vomiting, and diarrhea. Even a weak tea made from the flowers has been reported as
causing these symptoms in adult humans. Eating a great deal guarantees prolonged misery
and possibly more serious complications. Small children should be fenced away from this
plant. Clitoria ternate flowers, their characteristic blue color, and cyclotides, ultra-stable
macrocyclic peptides that are present in all tissues of this plant.

The latter are potent insecticidal molecules and are implicated as the bioactive agents
in a plant extract used commercially as an insecticide. To analyze cyclotide content, a single-
seed descent plant was grown for each of the 23 butterfly pea accessions. For each plant, two
similar but distinct compound leaves were separately harvested as technical replicates.
Peptide extraction was conducted by adding 2 mL of 50% acetonitrile (MeCN) in 1% formic
acid to every 100 mg of ground fresh sample. The samples were vortexed for one hour and
left to stand overnight at room temperature. The following day, they were centrifuged for 15
minutes at 13,300 rpm, and the peptide-containing supernatant was collected for LC–MS
analysis. Before analysis, the peptide extracts for each butterfly pea accession were spiked
with an internal standard. The ratio used was 30 μL of internal standard control peptide (5
μM kalata B2), 30 μL of extracted sample, and 10 μL of 50% MeCN. Sample injections (10
μL) were separated using a linear MeCN gradient at a flow rate of 0.25 mL/min on a
ZORBAX Rapid Resolution High Definition C18 UPLC column (150 × 2.0 mm, Agilent
Technologies, Santa Clara, CA, USA), maintained at 40°C and interfaced with a TripleTOF
5600 LC–MS (SCIEX, Ontario, Canada). The ionspray voltage was set to 5000 V, and the
source temperature to 500°C. LC–MS data were analyzed using MultiQuant™ 2.1.1 and
MarkerView™ 1.3 (SCIEX, Ontario, Canada). Signal intensities from the extracted ion
chromatograms were calculated based on the average area of the monoisotopic mass for
individual peptides, relative to the signal intensity of the kalata B2 internal standard (kB2,
3192 Da). Peptide profiling was performed twice on separate occasions for each accession to
ensure reliability according to (Oguis et al., 2020).
2.2. Red Fire Ant

Fire ants, particularly Solenopsis invicta (red imported fire ants, RIFAs), have
established themselves as globally invasive species with significant ecological and economic
impacts. These ants have been introduced to every continent except Antarctica and numerous
islands, posing a major threat to biodiversity and infrastructure (Wylie et al. 2019). Their
invasions may also have an impact on arthropod communities and have shown serious
detrimental effects on vertebrate animals, particularly ground-nesting birds and herpetofauna.
(King 2020a; Epperson et al. 2021). Because islands' ecosystems tend to be more delicate and
their species have fewer defenses against alien predators, the effects on wildlife are more
alarming.

Even at low populations, fire ants can have an impact on wildlife, in contrast to many
nonvenomous invading ants. One colony has the power to adversely affect neighboring eggs
and juveniles, causing major disturbances in population stability and reproductive
performance (Allen et al. 2017). Fire ants have long been considered a threat in North,
Central, and South America, but their growing widespread in the Asia-Pacific area makes
them an even more significant global issue (Gruber et al. 2017, 2021; Wylie et al. 2019).

Solenopsis invicta is a very versatile species that may live in urban, agricultural, and

natural settings. It is a generalist forager that is aggressive and able to establish dense

populations that control the majority of food sources. The strong invasiveness of the species

is a result of its capacity for quick reproduction and dissemination, effective colony

relocation, and exploitation of human disruptions (Gunawardana 2022). In addition, S. It has

been shown that invicta can build nests in both indoor and outdoor electrical equipment, such

as irrigation systems, network cable converters, streetlights, and telephone switchboards. This

conduct presents serious dangers to public safety and the economy because of electrical short

circuits and equipment failures (Liu et al. 2021).

 Although a lot of research has been done on the ecological and infrastructure effects

of fire ants, little is known about possible mitigation techniques. A growing field of study is

the application of biochar, a result of the pyrolysis of organic materials and a common soil

supplement in agricultural environments. However, its effects on S. invicta populations

remain largely unknown (Jiantao Fu et al. 2024). Developing sustainable and efficient

strategies to manage fire ant invasions and lessen their damaging effects on human

infrastructure and biodiversity requires an understanding of these relationships.

2.2.1. Ant Control

Invasion by the red imported fire ant (Solenopsis invicta) has devastating effects on
native biodiversity, agriculture, and public health. The ant’s aggressive foraging behavior and
high reproductive capacity have facilitated the establishment of wild populations in nearly all
regions where it has been introduced. According to (Ujiyama and Tsuji, 2018), invasive ants
can severely impact native ecosystems, agricultural productivity, and human health. The red
imported fire ant, which is highly aggressive and capable of stinging humans, has caused an
increasing number of individuals to suffer the consequences of its stings. Clinical reactions to
fire ant stings vary from mild discomfort to life-threatening anaphylaxis, with surveys
indicating that 0.6% to 6% of those stung experience anaphylactic reactions. The
consequences of fire ant infestations extend beyond public health. Fire ants can interfere with
mechanical and electrical infrastructure, often causing short circuits that lead to the
malfunction of equipment such as traffic signals. The economic impact of fire ant invasions
has escalated alongside their range expansion. By the early 21st century, the annual costs of
control measures, medical treatments.

The red imported fire ant was unintentionally introduced to the United States
approximately 80 to 90 years ago, spreading rapidly across the southern regions of the
country. More recently, it has been introduced to other parts of the world, including the
Caribbean, Australia, China, Taiwan, Japan, South Korea, and New Zealand. Thanks to its
aggressive foraging behavior and reproductive capability, the fire ant has established wild
populations in all introduced regions except New Zealand, Japan, and South Korea. It is,
therefore, plausible that wild colonies may eventually appear in these and other currently
uninvaded regions, according to (Ujiyama and Tsuji, 2018).
 Ants continue to pose a significant problem for microirrigation systems in various
agricultural regions, as evidenced by frequent reports from growers attributing system
failures to ant activity. In Spain, for instance, over 3 million hectares are irrigated annually,
with drip irrigation accounting for more than 40% of agricultural water use and up to 80% in
certain Mediterranean provinces according to (Pedro and Sanchez, 2022). Consequently, ant-
related damage poses a substantial threat to the economies of these areas, making it
imperative to find effective solutions to this issue according to (Pedro and Sanchez, 2022).

2.2.2. Previous Study of Ant Control regarding Plant

The red imported fire ant (RIFA) is one of the most detrimental invasive species,
threatening native ecosystems, human health and economic activities worldwide. In the
quarantine zone of Taiwan, RIFA re-infestation frequently occurs despite the intensive
application of synthetic pesticides, making its control costly and ineffective. Thus, there is an
urgent need to identify eco-friendly and sustainable alternatives for controlling RIFA
populations. This study examined the efficacy and feasibility of planting herbal species for
RIFA control. Five herbal species, Tagetes lemmonii, Armoracia rusticana, Cymbopogon
citratus, Cymbopogon nardus and Chrysopogon zizanioides, were grown in a RIFA-infested
field with local weeds as controls. Bait and pitfall traps and RIFA-intruded plants were used
to compare the ant activity in the control fields and those containing herbal plants. We further
evaluated the RIFA repellent activity of the five herbal plants and their basal soil through
digging bioassays. Generally, the field surveys showed more ants and intruded plants in
control than the herbal groups; however, the significance varied based on the trap type and
plant species. The digging bioassays demonstrated that the aboveground parts of T.
lemmonii, C. nardus, C. citratus and the belowground parts of T. lemmonii, C. citratus and C.
zizanioides effectively repelled RIFA. The basal soil of T. lemmonii, C. citratus and C.
nardus also exhibited deterrent activity towards RIFA. Our results demonstrated that herbal
plants are eco-friendly, sustainable alternatives for controlling and preventing RIFA
infestation in severe infested and non-infested areas. Tsai, C. C., Hung, S. H., Lin, X. R., &
Huang, R. N. (2022). Herbal plants as alternatives for the management of the red imported
fire ant, Solenopsis invicta. Journal of Applied Entomology, 146(, 975-989.)

The rapid evolution of pesticide resistance and the risks pesticides pose to human and
ecosystem health call for sustainable agricultural practices. Biological control of pests is a
promising tool in which natural enemies regulate pest densities and reduce damage.
Biological control (e.g. providing natural enemies in the ecosystem) not only reduces the use
of pesticides and production costs but also helps to maintain local biodiversity. However, the
success of biological control depends on many factors, such as the environmental factors and
traits of the species involved. The citrus growers in China were pioneers in biological control
using ants centuries ago. Over time, these organisms have been used to control pests around
the world, such as Spodoptera exempta (Walk.) in Kenya, forest pests in Canada, cocoa pests
in Ghana, crop pests in Nigeria and many other pests in different countries. Despite the
gradual evolution over time in the use of ants against pests, a major challenge still is to
identify positive and negative ant–crop matches, using management to boost positive effects
(services) and decrease negative effects (disservices).

Ants are considered as natural enemies of arthropods because they are abundant
generalist predators. As predators, ants perform services on crops such as reducing pest
abundance and plant damage (e.g. lost leaf area, fruit and seed damage), leading to an
increase in crop yield. However, the role of ants in agriculture is not yet completely clear
because they can also cause disservice. For example, ants can spread pathogens, increase the
density of honeydew-producing pest species (e.g. mealybugs, soft scales, aphids, psyllids or
whiteflies among others), and reduce the abundance of other natural enemies and pollinators.
For instance, pollinators can detect and avoid flowers if ants are present, decreasing
pollination services and compromising fruit formation. Therefore, it is essential to understand
the net effects of ants on biological control. The main biotic traits driving the role of ants in
the biological control of arthropod pests are related to the biology of the species involved (i.e.
ants, pests and natural enemies). For instance, aggressive ant species, which are usually
abundant and/or large-bodied ant species, are expected to have a greater capacity to reduce
pest abundance, mainly non-honeydew-producing ones. This reduction can lead to a decrease
in plant damage and an increase in crop yield. By contrast, these large-bodied or aggressive
ants can also affect negatively the abundance and behavior of natural enemies of honeydew-
producing pests. The specialization and dispersal type (winged or wingless species) of these
natural enemies may be affected by ants due to the lack of elaborate defense mechanisms
(most non-parasitoid species) and locomotion capacity of these enemies, respectively.
Therefore, it can lead to an increase in honeydew-producing pests, often causing disservices
to agroecosystems. Besides biotic traits, other factors such as field size, crop system and ant
exposure time (i.e. experiment duration) might affect their ecosystem services on biological
control in agriculture. For instance, the effect of field size on pest density depends on the pest
and natural enemy biology but how field size as well crop system might interact with ant
services have rarely been evaluated. For instance, more conservative farming with less
intensive management (e.g. shaded crops) is expected to conserve or even increase ant
diversity and may reflect positively on ants’ services such as herbivore predation. Finally,
numerous studies have evaluated the effect of ant exclusion on pests and other natural enemy
densities, but the experiment duration may have contrasting effects on pest abundance and
consequently in plant damage and crop yield.

Despite some meta-analyses testing ant-plant protection interactions, these studies

focus on natural systems rather than agriculture ones. Moreover, with the ongoing increase in
the conversion of natural systems into cultivated land [40], we need to understand how
biological and environmental traits drive these ant-plant outcomes in agricultural systems.
Here, our main goal was to review the role of ants in biological control, considering their
services and disservices. To do this, we evaluated the impact of ants on (i) pest abundance,
(ii) natural enemy abundance, (iii) plant damage and (iv) crop yield. We used a meta-analytic
approach, gathering information from published papers that compared the impacts of ant
presence in crops. We also evaluated whether ant effects were modulated by biotic traits,
namely: (1) body length of candidate ants (i.e. most abundant species); (2) pest type
(honeydew-producing species versus non-honeydew-producing species); (3) pest group
(taxonomic level of non-honeydew-producing species); (4) natural enemy specialization
(specialist versus generalist) and (5) natural enemy dispersal type (winged versus wingless).
In addition, we evaluated whether ant effects were modulated by: (6) field size; (7) crop
system (monoculture, intercropped or shaded crops) and (experiment duration (e.g. ant
exclusion experiment).

2.2.4. Ant Control Solution Production

Ant colony optimization is one of well-known techniques to solve the global multi-
objective optimization problem. In the article, the authors present a solution of the production
scheduling problem by means of an ant colony optimization algorithm. A case study of the
algorithm efficiency estimated against some others production scheduling algorithms is
presented. Advantages of the ant colony optimization algorithm and its beneficial effect on
the manufacturing process are provided.
2.2.5. Standard Ant Control Methods

The red imported fire ant (RIFA) (Solenopsis invicta) is recognized as one of the most
damaging pest species globally. Workers swarm in large numbers when their nests are
disturbed, delivering stings that cause wheal-and-flask reactions with sterile pustules. These
stings result in persistent pain and itching, lasting for weeks, and may lead to complications
such as inflammation, secondary infections, anaphylactic shock, or even death. RIFA attacks
occur both indoors and outdoors, posing significant risks to vulnerable individuals such as
infants and immobilized patients. In 2011, the medical costs of RIFA-related incidents in
Australia reached AUD 114 million, and developing Pacific Island nations are projected to
face medical expenditures of up to USD 35.1 million if invaded by RIFA according to
(Gruber et al., 2021). RIFA management predominantly relies on synthetic insecticides like
pyrethroids (cypermethrin, cyhalothrin), nicotinoids (imidacloprid), indoxacarb, and growth
regulators (pyriproxyfen, methoprene, diflubenzuron). While effective, these chemicals have
several drawbacks, including harm to non-target beneficial species, the need for frequent
application, and the potential for bioaccumulation, which can disrupt ecosystems by causing
the local extinction of key species according to (Fu et al., 2018).

To reduce reliance on synthetic pesticides, sustainable approaches such as the use of

natural enemies (e.g., the phorid fly-borne pathogen Kneallhazia solenopsae and competitive
ant species Monomorium minimum), as well as advanced aerial surveillance systems, have
been proposed according to (Spring et al., 2017). Among alternative strategies, plant-based
repellents are particularly appealing due to their eco-friendliness. However, many plant-
derived repellents are volatile and require specialized formulations to prolong their effects.
Live herbal plants, on the other hand, continuously produce bioactive compounds, offering a
sustainable alternative according to (Chen and Oi, 2020). To examine the feasibility of using
herbal plants for RIFA control, five species (Tagetes lemmonii, Armoracia rusticana,
Cymbopogon citratus, Cymbopogon nardus, and Chrysopogon zizanioides) were planted in a
RIFA-infested field, with local weeds serving as controls. Ant activity and plant intrusion
were assessed using bait and pitfall traps. Results indicated lower ant activity and fewer
intruded plants in fields with herbal plants compared to control fields, though significance
varied by plant species and trap type.
 Digging bioassays further confirmed the repellency of the aboveground parts of T.
lemmonii, C. citratus, and C. nardus and the belowground parts of T. lemmonii, C. citratus,
and C. zizanioides. Additionally, the basal soil of T. lemmonii, C. citratus, and C. nardus
exhibited deterrent activity according to (Tsai et al., 2021). While the initial cost of planting
herbal species may seem high, the long-lasting repellent effects make them a cost-effective
solution in the long term. Carefully selected plants should be robust, resilient, and capable of
outcompeting weeds, requiring minimal maintenance once established. Additionally, plants
with added economic value—such as citronella, lemongrass, vetiver, and marigold, which
can be used for essential oils or as condiments—may increase landowners’ willingness to
invest in their cultivation according to (Tsai et al., 2021).

Medicinal plant extracts have been widely recognized for their biological activity
against various insect species according to (Eva et al., 2017; Singh and Pandey, 2018;
Noaman and Bahreininejad, 2024). These extracts hold promise for development into novel,
natural insecticides. However, there is limited research on the biological activity of these
extracts specifically against red fire ants (Solenopsis invicta). Previous studies have
demonstrated that medicinal plants from families such as Labiatae, Umbelliferae,
Zingiberaceae, and Neemaceae exhibit significant bioactivity against various pests.
According to (Guo et al., 2025) reported that crude extracts derived from 10 plant sources
displayed varying degrees of contact toxicity toward S. invicta, indicating their potential as
natural control agents despite differences in efficacy. Other research has supported the
potential of plant extracts as sustainable pest management tools according to (Aminu Sulhath
et al., 2024; Pietro et al., 2022; Mohammad et al., 2024). This highlights the biological
activity of plant-derived crude extracts against S. invicta, further expanding the application of
botanical solutions in pest control. Plant extracts demonstrate not only significant pest control
activity but also environmental sustainability. Exploring extracts with potent repellent or
toxic effects on pests can provide a theoretical foundation for developing green,
environmentally friendly insecticides according to (Guo et al., 2025).

2.2.6. Alternative Methods of Ant Control

1. Given the environmental issues mentioned above, alternate ant control techniques,
characterized as having little to no effect on species that are not the target, have gained more
attention, with a focus on using "natural" remedies that are said to be repellant and
insecticidal action, such as hydrogels; essential oils as insecticides and repellents and
pheromones, both by themselves and in combination with other goods. Evaluations of
substitute Argentine ant pest control methods have already been published.

2. In urban and suburban settings, ants (Hymenoptera: Formicidae), particularly the

Argentine ant, Linepithema humile (Mayr), can be serious nuisance pests. The mainstay of
conventional interventions has been the application of chemical insecticides, specifically
bifenthrin and fipronil, as residual contact treatments around the exterior of infested
structures. Significant water contamination results from the continued reliance on insecticides
to control ants, even though stricter regulations are restricting the range of insecticide
applications in urban areas. The U.S. EPA has further limited the ways in which many ant-
control insecticides are used in both professional and over-the-counter markets. This review's
goal is to provide an overview of the pertinent research on managing nuisance pest ants, with
a focus on L. modest, concentrating on low-impact, alternative (to broadcast applications)
pest control techniques and avoiding the use of liquid broadcast applications of EPA-
registered insecticides. RNA Interference (RNAi), Food Source Reduction, Hydrogels,
"Virtual" Baiting, and Extremely Low Bait Concentrations, Mass Trapping, Trail Pheromone,
Use of Behavior-Modifying Chemicals, and Deterrents are some of the specific subsections.
 3. METHODOLOGY

3.1 Research design

3.2 Research setting

3.3. Research instruments

3.4. Data Gathering Procedure

3.5 Data Analysis

 LITERATURE CITED

Aminu, T.A. Sulhath, Visakh, N.U. Pathrose, B. George, S.B. (2024) Investigating the
insecticidal properties of essential oils extracted from wild turmeric (Curcuma
aromatica salisb) leaves waste against three key stored product pests Sustainable Chem.
Pharm., 38, Article 101482. Available at: 10.1016/j.scp.2024.101482

Anjos, D. V., Tena, A., Viana-Junior, A. B., Carvalho, R. L., Torezan-Silingardi, H., Del-
Claro, K., & Perfecto, I. (2022). The effects of ants on pest control: a meta-analysis.
Proceedings of the Royal Society B, 289(1981), 20221316.

Allen, C. R., Epperson, D. M., & Garmestani, A. S. (2004). Red imported fire ant impacts on
wildlife: a decade of research. The American Midland Naturalist, 152(1), 88-103.

Chen, J. Oi, D.H. (2020) Naturally occurring compounds/materials as alternatives to synthetic

chemical insecticides for use in fire ant management. Insects 11:1–18. Available at:
https://doi.org/10.3390/insects11110758

Chernigovskiy, A. S., Kapulin, D. V., Noskova, E. E., Yamskikh, T. N., & Tsarev, R. Y.
(2017, October). Production scheduling with ant colony optimization. In IOP
Conference Series: Earth and Environmental Science (Vol. 87, No. 6, p. 062002). IOP
Publishing.

Damalas, C. A., & Koutroubas, S. D. (2018). Current status and recent developments in
biopesticide use. Agriculture, 8(1), 13.

De Pedro, L., & Sanchez, J. A. (2022). Natural Repellents as a Method of Preventing Ant
Damage to Microirrigation Systems. Insects, 13(4), 395.
https://doi.org/10.3390/insects13040395

Epperson, D. M., Allen, C. R., & Hogan, K. F. E. (2021). Red Imported Fire Ants Reduce
Invertebrate Abundance, Richness, and Diversity in Gopher Tortoise Burrows.
Diversity, 13(1), 7. https://doi.org/10.3390/d13010007

Eva, M.G. Nolwenn, D. Naresh, S. Jürgen, G. (2017) The potential of medicinal and aromatic
plants (MAPs) to reduce crop damages by Asian Elephants (Elephas maximus) Crop
Prot., 29–37, pp. 0261-2194. Available at: 10.1016/j.cropro.2017.06.002

Fu, H. Hardy, J. Duff, K.E. (2018) Selective vulnerability in neurodegenerative diseases.

Available at: https://www.nature.com/articles/s41593-018-0221-2

Fu J, Qin M, Liang Y, Lu Y, An Y, Luo Y. Toxicity and Behavioral Effects of Amending

Soils with Biochar on Red Imported Fire Ants, Solenopsis invicta. Insects. 2024 Jan
8;15(1):42. doi: 10.3390/insects15010042. PMID: 38249048; PMCID: PMC10816398.
Gruber, N. Boyd, P.W. Frölicher, T.L. Vogt M. (2021) Biogeochemical extremes and
compound events in the ocean. Available at: https://www.nature.com/articles/s41586-
021-03981-7

Guo, Y. Liu, Z. Ye, Y. Chen, Y. Wu, Z. (2025) Antifeedant and contact toxicity activity of
crude extracts from 10 plants to red imported fire ant, Solenopsis invicta Buren
(Hymenoptera:
Formicidae).Availableat:https://www.sciencedirect.com/science/article/pii/
S1226861524001559

Gunawardana,D,cabicompendium.50569,CABICompendium,doi:10.1079/
cabicompendium.50569, CABI International, Solenopsis invicta (red imported fire ant),
(2022)

King, J. R. 2020a. Fire ants (Solenopsis, in part). Pages xx-x in C. Starr, editor. Encyclopedia
of Social Insects. Springer, Cham, Switzerland.

Oguis, G. K., Gilding, E. K., Jackson, M. A., & Craik, D. J. (2019). Butterfly Pea (Clitoria
ternatea), a Cyclotide-Bearing Plant With Applications in Agriculture and Medicine.
Frontiers in Plant Science, 10. https://doi.org/10.3389/fpls.2019.00645 Oram, R. N.
(1992). Register of Australian herbage plant cultivars. Aust. J. Exp. Agric. 32, 547–
548.

Liu, Y.S.; Huang, S.A.; Lin, I.L.; Lin, C.C.; Lai, H.K.; Yang, C.H.; Huang, R.N.
Establishment and social impacts of the red imported fire ant, Solenopsis invicta
(Hymenoptera: Formicidae) in Taiwan. Int. J. Environ. Res. Public Health 2021, 18,
5055. [CrossRef] [PubMed]

Mohammad, H. Amin, S. Reza, N. Mohsen, M.D. Hojjatollah, M. Maryamossadat, N.M.

Mahalat, M.-M. Roya, A. (2024) Biological activity of essential oils from Ferulago
angulata and Ferula assa-foetida against food-related microorganisms (antimicrobial)
and Ephestia kuehniella as a storage pest (insecticidal); an in vitro and in silico study
Fitoterapia., 175, Article 105937. Available at: 10.1016/j.fitote.2024.105937

Noaman, V. Bahreininejad, B. (2024) Evaluation of the acaricide activity of extracts from

eight species of medicinal plants on larvae of Hyalomma anatolicum (Acari: Ixodidae)
ticks J. Asia-Pac. Entomol., 27 (2), Article 102231. Available at:
10.1016/j.aspen.2024.102231

Oguis, G. K., Gilding, E. K., Huang, Y. H., Poth, A. G., Jackson, M. A., & Craik, D. J.
(2020). Insecticidal diversity of butterfly pea (Clitoria ternatea) accessions. Industrial
Crops and Products, 147, 112214.

Pietro, F. Domenico, R. Domenico, C. Mauro, M. Gian, C.M. Sergio, G. Aldo, T. (2022)

Composition and biological activity of essential oils from Artemisia roxburghiana
Besser and Elsholtzia fruticosa Rehder cultivated in Italy Ind. Crops Prod., 187, Article
115317. Available at: 10.1016/j.indcrop.2022.115317
Spring, D. Croft, L. Kompas, T. (2017) Look before you treat: increasing the cost
effectiveness of eradication programs with aerial surveillance. Biol Invasions 19:521–
535. Available at: https://doi.org/10.1007/s10530-016-1292-1

Singh, P. Pandey, A.K. (2018) Prospective of essential oils of the genus Mentha as
biopesticides: a review Front. Plant Sci., 1295. Available at: 10.3389/fpls.2018.01295

Suarna, I. W., & Wijaya, I. M. S. (2021). Butterfly Pea (<i>Clitoria ternatea</i> L.:
Fabaceae) and Its Morphological Variations in Bali. Journal of Tropical Biodiversity
and Biotechnology, 6(2), 63013. https://doi.org/10.22146/jtbb.63013

Suiter, D. R., Gochnour, B. M., Holloway, J. B., & Vail, K. M. (2021). Alternative methods
of ant (Hymenoptera: Formicidae) control with emphasis on the Argentine ant,
Linepithema humile. Insects, 12(6), 487.

Tsai, C. C., Hung, S. H., Lin, X. R., & Huang, R. N. (2022). Herbal plants as alternatives for
the management of the red imported fire ant, Solenopsis invicta. Journal of Applied
Entomology, 146, 975-989.

Tsai, C. Hung, S. Lin, X.R. Huang, R.N. (2021) Herbal Plants as Alternatives for the
Management of the Red Imported Fire Ant, Solenopsis Invicta (Hymenoptera:
Formicidae). Available at: https://www.researchsquare.com/article/rs-676405/v1

Ujiyama, S., & Tsuji, K. (2018). Controlling invasive ant species: a theoretical strategy for
efficient monitoring in the early stage of invasion. Scientific Reports, 8(1), 8033.
Wylie, R., Yang, C. C. S., & Tsuji, K. (2020). Invader at the gate: The status of red imported
fire
ant in Australia and Asia. Ecological Research, 35(1), 6-16.