Fumigant Repellency of Oleic Acid on

Periplaneta Americana (American Cockroaches)

Jhonloyd G. Batralo1, Neil Ivan N. Briones2, Althea Jade G. Condes3, Casselyn A. Medrana4,
Jaron Medrana5, Renz Aldwin U. Montealegre6, Genesis E. Perez7, Jet Nathaniel M. Pitogo8, Elisa
Samantha C. Sta. Ana9, Janna Mekylla U. Villegas10
ABSTRACT
Oleic acid has been shown to be the necromone for a wide variety of insects. Studies suggest that
this chemical is responsible for necrophobic behavior in most cockroach species. Contrary to early
research, anecdotal statements from most pest control blog websites argue that oleic acid attracts American
cockroaches rather than repels them. This research aims to validate which side is refutable by testing the
fumigant repellency of pure oleic acid on American cockroaches and whether sex and food factors affect
the subjects' behavior using a choice box divided into three chambers (options): Feeding (FC), Treatment
(TC) and Repel Chamber (RC) with the latter two further divided to 5 levels. Each mark has 10 cm
increments which were used to measure the mean range of the chemicals. Two concentrations (30 & 60%)
were diluted in acetone to test if concentration affects general repellency. Results show an inverse
proportion between concentration and repellency percentage as the 30% group induced a high repellent
activity (95%), while the 60% and 100% group showed lower (87.67% & 68.33% respectively). Sex and
food factors did not show significant effects on the repellency of oleic acid solutions (sex: p=0.104 & food:
p=0.742). Apparently, the negative control also showed high repellent activity of 90% which was
hypothesized to be linked to its uncertain purity. The results indicate that pure oleic acid is more effective
on contact than it is from a distance due to its low volatility.

INTRODUCTION bacterias, viruses and other harmful

Most organisms have their function and
ecological value. Cockroaches for example are
biological recyclers, they help break down
organic materials and fertilize the soil (Yusuf,
2022). Although most are useful in agriculture,
some species of cockroaches have adapted to
households which is what classified them as
pests. The American cockroach, Periplaneta
Americana (Linnaean) is among the four
house-infesting species of its kind (Britanicca,
2022;Hahn and Ascerno, 2019) and is regarded
as the most important invasive urban pest
(Nasirian and Salehzadeh, 2018). Moreover, the
Periplaneta Americana is one of the two most
abundant species of roaches in local households
and infrastructures (i.e. restaurants, groceries,
etc.) which poses a threat to (local food
processing franchises and) overall public health
in general (Labayog, 2019).
American cockroaches naturally occur
outdoors particularly in sewers (Nasirian and
Salehzadeh, 2019) and only enter houses in
search of food (Barbara, 2000/2021). Their diet
consists of almost anything organic including
decaying materials, glue, grease, leather,
bookbindings, and sometimes even the carcass of
their fellows (Hadley, 2019). They also feed on
human food and are vectors of different
microorganisms that they usually get from their
origins: sewers. According to the World Health
Organization (1999/2020) some of the common
illnesses linked to these pathogens are infections
on the urinary tract, gastroenteritis, salmonella
infections and asthma attacks (Poowuttikul et al.,
2019) in some cases. Due to its medical and
health significance, American cockroaches have
been the subject of pest control research for
decades.
Efforts on preventing infestations of
American cockroaches and other species are
well-documented. Chemicals such as boric acid
(Luna, 2019), emulsifiable concentrate, carbaryl,
diazinon, chlorpyrifos and fog formulations
(Zahraei-Ramazani et al., 2018) are among the
commonly used synthetic chemicals for pest
management. However, multiple studies also
suggest that the use of these synthetic or
man-made chemicals promotes tolerance on
American cockroaches. For instance, a study on
American cockroaches chemical resistance
conducted by Zheng et al. (2022) posits that the
genes of American cockroaches associated with
detoxification were significantly expanded which
links to pesticide resistance of the species.
Likewise, Tahir et al. (2017) also studied
resistance of the said species against malathion

1
in which results suggest that a higher activity of
insecticide detoxifying enzymes was observable.
Furthermore, Rahimian et al. (2019) suggested
that Periplaneta Americana had fair levels of
resistance against different insecticides. These
findings suggest that toxic chemicals may
backfire in the long run for building tolerance.
Therefore, it may be beneficial to utilize natural
and organic chemicals found in the species
themselves in order to cease their rapid
adaptation by triggering the natural evasion
behavior of the species.
Now, the Periplaneta Americana and
generally all species of cockroaches have
sophisticated senses that can detect certain
chemicals. One of the chemicals that
cockroaches and most insects in general are
sensitive to is called oleic acid, a fatty acid
secreted by their dead counterparts (Shibao et al.,
2022). According to an old study by Rollo et al.
(1994), the universal "death stench" for a wide
variety of insects, including roaches, is oleic acid
– a compound that many insects are repelled by
to avoid what caused the death of their comrades.
However, succeeding literature on cockroaches'
response to the substance asserts the opposite.
The idea that cockroaches are attracted to the
compound released by the carcass of their
fellows is almost verbatimly stated in most pest
control blog websites. However, these blog
websites failed to present research sources of
their information nor datas that supports their
claim; These claims are mostly based on
anecdotal information. Conversely, some
literature still supports a counter argument for
this attractive perspective. A literature by
Andersen (2022) supports the initial findings by
Rollo et al. (1994) that the release of oleic acid
upon death is a warning that predators or harmful
chemicals must be around and they will avoid
the carcasses unless they are starving.
Nonetheless, he was also unable to cite sources
other than the original findings. Furthermore,
multiple researchers have also tested oleic acid
on other insects such as on honey bees: triggered
hygienic response and corpse removal (McAfee
et al., 2018); on ants: burial and cannibalism of
treated subjects (Sun et al., 2018); and on
crickets: repelled by the scent (Aksenov and
Rollo, 2017).
With all the accumulated contrasting
findings on this topic, this compound and its
effects on the behavior of multiple insects
particularly on American cockroaches remains
vague (Domingue et al., 2020). The current
research is concerned about the interaction
between Periplaneta Americana and oleic acid
and the substance's potential to be used as a
cockroach repellent. To make this possible, oleic
acid should be managed and spread effortlessly
within less concentrations. One way to achieve
such is by mixing it with a chemical capable of
dissolving and carrying it. Acetone is a widely
used chemical in dispersing chemicals such as
essential oils for repellency testing not only
limited to research on cockroaches but also on a
wide variety of insects. Additionally, most
research suggests that acetone elicits low to zero
percent repellency against American
cockroaches when used as negative control
treatment (Ngoh et al, 1998; Appel et al., 2001;
Schultz et al., 2004; Omara et al., 2013).
Therefore, it may serve as a carrier without
overwhelming the effect of the active chemical.
OBJECTIVES OF THE STUDY
The objective of the current study
includes two major aims: (1) To discover the
effect of oleic acid on American cockroaches'
behavior in terms of aggregation or repulsion in
order (2) To create a product that effectively
utilizes the interaction between the substance and
the species of concern. This study intends to
monitor how cockroaches respond to oleic acid –
whether they are attracted to or repelled by it –
and leverage their response. More specifically,
this research aims to provide answers for the
following topic under question:
I. General Effect of Substance
To discover the relationship between
oleic acid and the behavior of American
Cockroaches towards it; Whether the findings
will support or oppose the initial research in the
topic. The research also seek answer to the
following sub-topics:
1. Sex as factor
Aside from the effect of the substance
on the subjects in general, the specificity of the
results with respect to the sex of the subjects will
also be assessed; To discover whether the
substance significantly affects only a particular
sex group.
2. Food as factor
To test whether the discriminatory
conditions regarding food supply and availability
significantly affects the behavior of the subjects
towards oleic acid.
2

II. Oleic Acid Repellent Spray
To produce a repellent utilizing the
influence of oleic acid on American cockroaches'
behavior and test the potential product in terms
of:
1. Concentration
The study will examine the
concentration ratio demands of the oleic acid to
acetone for it to be, if it will be, an effective
repellent. The different concentration will be
diluted in acetone using conventional mixing.
2. Feasibility
The oleic acid repellent will be tested
under controlled conditions, and if elicited a
positive result, will subsequently be followed by
tests under natural conditions to determine its
feasibility of use in practical situations. The
experiment will use ready-made containers with
dimensions enough for interaction between
cockroaches to cockroaches, cockroaches to the
environment and cockroaches to oleic acid.
3. Overall Efficacy
The product will be tested in terms of
its capability on doing its supposed purpose – to
repel American cockroaches. The effect of the
oleic acid solution will be assessed in terms of
overall efficacy.
SCOPE, DELIMITATION AND
LIMITATION OF THE STUDY
This study will focus on American cockroaches,
Periplaneta Americana. The age of the subjects
will be limited to their adult life stage only as
they can be easily observed and gathered
compared to younger conspecifics. The subjects
will include both male and female American
cockroaches. They will be evenly distributed to
designated containers specific to their sex. The
major limitation of the current study is the
procuring of chemicals and achieving the subject
population demands of the study as they were
caught from common households due to
unavailability of laboratory reared cockroaches
locally.
Another limitation that the study may
encounter is the complexity of other
environment-present factors that may affect the
results of the interaction. This includes some
variables that require specific equipment to
control, modify and measure such as humidity,
light intensity, moisture and temperature.
SIGNIFICANCE OF THE STUDY
This research will be conducted to
provide crucial information and new insights
about the chosen topic - whether the findings
will support or oppose the initial knowledge on
the topic.
Through this study, the pest
management community will further understand
how they can utilize the chemical that
cockroaches and possibly other insects release
upon death.
Additionally, by helping the pest
management community, the results of the study
may help promote public health and reduce cases
of infestations from American cockroaches
which results in multiple disease transmission.
Furthermore, the findings of this study
may provide the framework for further research
into the potential repelling or attracting
properties of oleic acid on American
cockroaches.
DEFINITION OF TERMS
American Cockroach (Periplaneta
americana). A species of cockroach that
originated from South America. Typically at
30-40 mm with a Reddish-brown color
(Zahraei-Ramazani et al., 2018).
Pest. Invasive organisms, particularly of
the insect kingdom, are capable of significant
damage to agriculture and biodiversity
(Epanchin-Niell et al. 2021).
Oleic acid. A kind of monounsaturated
fatty acid that is frequently present in olive oil
and also appears in other vegetable-type oils and
animal goods (Tutunchi et al., 2020).
Fatty acid. Consists of a carbon chain
with oppositely placed methyl and carboxyl
groups. Saturated and unsaturated fatty acids are
the two categories of Fatty acids (Lund and
Rustan, 2020).
Insect Repellent. A compound used for
repelling insects such as mosquitoes, and be used
as protection or preventative for mosquito
3
borne-illnesses like malaria or dengue (Ling Lo
et al., 2018).
RESEARCH METHODOLOGY
A quantitative experimental design was
followed. Research subjects (Periplaneta
Americana) and the environment for experiment
were assessed for different factors that may pose
underlying effects on the results of the study.
Moreover, the study randomly assigned the
subjects to sets of experimental and control
groups.
Test Insects
Approximately 60 adult American
cockroaches were caught from common
households and sewers using a trap mechanism
and manual catching. The subjects were reared in
an aquarium (61 x 30.5 x 31 cm) provided with
pellet Pedigree Beef pro and mineral drinking
water as main foraging sources, and cartons as
harborages; Food and water sources were
replaced ad libitum. A carton covered with
plastic was attached to the rearing container as
the lid and was randomly poked to prevent the
subjects from suffocating. The aquarium was
placed on a high table to avoid ant invasions.
Dead cockroaches were discarded as soon as
they showed behaviors related to death (e.g.
lying flat on their back, weak reaction to
antennal tactile stimulation) to avoid infections
from spreading within the culture. The subjects
were randomly assigned to a set of positive
control, negative control and three experimental
sets of groups.
Test Chemicals and Apparatus Used
The test chemicals were purchased from
verified online chemical shops. One liter of Oleic
acid was purchased from Dalkem Corporation.
The substance was kept in its packaging before
and after use to avoid external conditions from
deteriorating the chemical. In search of pure
acetone for the solution solvent, 4 bottles of
Wins Cloud Premium Acetone were bought from
Serna Brands online shop which the seller
testified to be 100% Acetone. Meanwhile, 50 ml
of Clove Oil was purchased from scentfullyyours
online shop. Clove oil was chosen as the positive
control for its proven repellent effect against the
insects under question (Omara et al., 2013).
Moreover, the laboratory tools and
equipment were also bought from online sources.
Two 10 ml pipettes were purchased from Eshop
Beauty & Bottles cosmetic shop while the
beaker, glass stirrer, wash bottle, and filter paper
were bought from The Lab Store online shop. On
the other hand, the petri dish was donated by the
School Laboratory of Tanauan Institute
Incorporated. Moreover, protective garments
such as rubber gloves and KN95 face masks
were issued by the local Health Center of
Barangay Pantay Matanda Tanauan City through
a verbal request.
Experiment Container
Essentially, the container design of the
current study followed the concept of Yoon et al.
(2009) wherein the test insects were given two
options: either to enter the treated chamber or the
untreated ones which corresponds to the effect of
the substance to the subjects' behavior. Four
identical containers primarily made of plastic
sheets supported by wood beams with openable
lid were used in the experiments. Five identical
holes were poked in the lid of the containers to
allow the respiration of the cockroaches.
Moreover, the container was divided into three
sections namely the Treatment chamber (TC) and
Repel chamber (RC) on either side, and the
Feeding chamber (FC) in between the two, each
serving distinct purposes. The dimensions
(Length x width x height) and appearance of
each chambers are as follows (Fig.1):
Experiment Container (Fig. 1)
The floor and wood beams were all covered by the plastic.
The measurements of each chamber was based on the area
covered by the plastic – the wood beams were not factored as
part of the chamber dimension.
● Treatment chamber (50 x 20 x 20 cm)
● Feeding chamber (15 x 20 x 20 cm)
● Repel chamber (50 x 20 x 20 cm)
Each container is linked by two
identical sliding doors with 12 cm wide and 4 cm
high openings. The floors of both the Treatment
4
and Repel chambers were marked with thin black
tape dividing the chambers into 5 regions with
10 cm spaces. These regions are referred to as
levels 1 to 5 with respect to the wall opposite to
the sliding door or the side where the treatment
will be placed in the TC (farthest: 1 ; nearest: 5).
The indicated levels were used as a basis to
measure how attracted or repelled the subjects
were. Furthermore, the containers were situated
in an experiment room walled with black mantle
to maximize control over lighting conditions.
Repellency Test
The repellency of Oleic acid solutions
were tested against adult male and female
American cockroaches in separate simultaneous
treatments. Subjects with intact antennae were
distributed to 4 groups specific to their sex (2
male groups and 2 female groups) corresponding
to 4 individual experiment containers which
constitute a single set. Each set has a male and a
female group provided with both food and water
while another male group and female group were
provided with water only. These sources were
placed in the Feeding chamber of each container.
(Table 1 - Subject Population and Distribution)
Two concentrations of the substance
(30% and 60% oleic acid) dissolved in acetone
were tested for repellency. A resulting volume of
5 ml was treated to a circular filter paper (The
Lab Store 90 mm) placed in a 90 mm petri dish.
Additionally, pure oleic acid was also tested for
repellency. The treatments were placed at the
outermost side of the treatment chamber of each
container. The cockroaches were first weighed
before transferring them to the experiment room.
They were acclimatized in their respective
container for a total of 48 hours (Reza et al.,
2010; Ubulom et al., 2021) maintaining a light
and dark period of 12h:12h (Babri et al. 2012).
During this phase, the sliding doors are opened
allowing the cockroaches to access all parts of
the container. Before the treatment phase at the
end of the 48 hour familiarization, the subjects
were transferred to individual tupperwares to
weigh their post-acclimatization mass.
Afterwards, the cockroaches were enclosed to
the Feeding chamber while keeping the sliding
doors closed. Then, the treatment is placed in its
chamber followed by a diffusion period of the
substance. The time of diffusion was calculated
based on the amount (in ml) of acetone in the
solutions following the prior work by Kamaldin
and Jaal (2018) in which they stated that the
evaporation rate of acetone on filter paper is
116.3 mg/min. Since 1 ml of acetone equates to
785 mg, the diffusion time was solved using the
formula:
Diffusion Time = acetone (ml) × 785mg/ml
116.3mg/min
After the allotted time for chemical
accumulation, the sliding doors were opened
simultaneously. Moreover, since the onset of the
familiarization phase began with a dark period,
the treatment was also held during a dark period,
particularly the third. To account for the dark
period, four identical carton boxes were
assembled such that it blocked the light while
exposing one side of the container through which
an observer can see the subjects inside; The
lights are not turned off during the dark period
but are rather blocked by the cartons creating a
dark ambience from the shadow. This setup is
ideal to prevent the effects of light on the
behavior of the cockroaches without jeopardizing
the observability of the experiment. Furthermore,
after the exposure of the treatment substance, the
subjects' behavior was recorded for every 30
minutes for the first 2 hours followed by an 8
hour-long exposure. As such, the cockroaches
were exposed to the substances for a total of 10
hours. The location of each cockroach was
recorded during the 30 mins, 60 mins, 90 mins,
120 mins, and 10 hour mark. The repellency of
the substances were calculated using the formula
as applied by Ubulom et al. (2021) which were
modified to match the context of the current
study:
5
 Repellency (%) = 100 - T × 100 RESULTS
N
where
Repellency
T = the number of visits to the
The repellent activity of the oleic acid
Treatment Chamber.
solutions was tested against male and female
N = the summation of all visits to all
adult American cockroaches. An inverse
chambers.
proportion between oleic acid concentration and
repellency was observed (Table A) from the
Experimental Groups
gathered data analyzed using Two-way ANOVA
As mentioned earlier, three
with the help of JAMOVI online statistical
concentrations of oleic acid (30, 60, and 100%)
software. Among the three concentrations, the
will be tested for repellency in separate
30% oleic acid group was recorded to have the
experiments. The experimental sets were
highest repellency. Table B shows that both the
grouped into four. (A1 & B1 are) A group of
30% and 60% groups showed no significant
males with food and without food, and a group
difference compared to the positive control,
of females with food and without food. Distinct
p=0.999 and p=0.182 respectively, indicating a
groups were treated with either 30%, 60% or
repellent activity comparable to that of the Clove
100% of oleic acid with a final volume of 5 ml
oil. Apparently, the negative control groups also
per treatment.
showed no difference when compared to the
positive control (p=0.901) which suggests that
Control Groups
both Clove oil and acetone induced a repellent
Each container holds 3 cockroaches of a
effect on the subjects. On the other hand, the
particular sex group and food availability
100% oleic acid group showed a significant
condition. Similar to the experimental groups,
difference when compared to both the 30% oleic
the Positive control included four groups: males
acid group (p=0.005) and positive control group
and females provided with food and water, and a
(p=0.003) indicating a less repellent activity
male and female group respectively provided
compared to the latter two.
with water only. The positive controls were
treated with 2 ml Clove oil mixed with 3 ml Table A. Overall Repellency (%) of test chemicals against
acetone arriving at a solution with 40% Clove oil American cockroaches
concentration. Similarly, the Negative control
was grouped into four namely the C1 & C2: male
and female respectively provided with food and
water, and C3 & C4: male and female
respectively provided with water only. The
negative controls were exposed to 5 ml pure
acetone smeared in a filter paper.

Statistical Analysis
Both One-way and Two-way Analysis
of Variance (ANOVA) were utilized to analyze
the collected data and test the hypothesis of the The repellency (%) was calculated using the same formula
current study. The analyses were conducted in used by Ubulom et al. (2021). Repellency is defined as the
number of visits to Feeding Chamber and Repel Chamber.
the JAMOVI statistical software version 2.3.25.

Additionally, a System Usability Survey

was utilized to take the insights of 30 Grade 12
STEM students from Tanauan Institute Inc. about
the acceptability of the potential product based
on its general characteristics. Four factors
namely the Production/Market Cost,
Environmental-User friendliness, Materials used
and Intended Purpose were discussed in the
survey. The responses were analyzed using
Microsoft Excel version 14.0.4734.1000 (32-bit)
in Windows 10.

6
Table B. Tukey HSD Post Hoc Test.
Repellency comparison between test chemicals based on number of FC and RC
female group treated with 60% oleic acid
visits. (p=0.014); Lastly, the 60%-male group also
showed significant difference to 100%-male
group (p=0.031) (Table 1.5).
Concentration-wise, only the 30% group showed
difference to the 100% group (p=0.006) while
the 60% group showed no significant difference
to both the 30% group (p=0.173) and 100%
group (p=0.339) which implies that the former
lies between the latter two in terms of repellency
(Table 1.6). Nevertheless, the general
comparison between all oleic acid groups
showed that both sex and food factors
individually do not have a significant effect on
the repellency of the solutions (sex: p=0.104 ;
food: p=0.742) (Table 1.4).
Table 1.1. ANOVA. 30% Oleic acid group: Sex-Food factor

(*p<0.05, **p<0.01, ***p<0.001)

Sex and Food as factor Table 1.2. ANOVA. 60% Oleic acid group: Sex-Food factor
This study was conducted to determine
whether the difference in sex and food
availability conditions will have a significant
effect on the behavior of cockroaches towards
the oleic acid solutions. The hypothesis was
tested using Two-way ANOVA to discover if the
repellency of the substances varies according to
the conditions posed to the subjects. Results Table 1.3. ANOVA. 100% Oleic acid group: Sex-Food factor
from the Tukey's HSD Post Hoc analysis shows
that the sex specific grouping and food
availability did not have a significant effect on
the repellency of both 30% (sex: p=0.572 &
food: p=0.572) and 100% oleic acid solutions
(sex: p=0.565 ; food: p=0.565) as shown in
Table 1.1 and 1.3 respectively. Table 1.4. ANOVA. Oleic acid groups comparison.
Counterintuitively, results of the 60% oleic acid Effects of sex and food to the repellency of the oleic acid
shows that both factors have a significant effect solutions.
on the repellent activity of the concentration
(sex: p<0.001 ; food: p=0.44). An interaction
between the two factors was also recorded to be
significant in the 60% group (p=0.044) as shown
in Table 1.2. This difference also manifested in a
separate analysis between all oleic acid groups as
results show that there was an interaction
between the concentration of oleic acid and sex
of the subjects (p=0.011). Tukey's HSD Post Hoc
Test shows that there was a significant difference
between 30%-male groups and 60%-female
group (p=0.031); A significant repellency
difference was also recorded between male and

7
Table 1.5. Tukey's HSD Post Hoc Test.
Interaction between concentration and sex towards repellent
activity
Table 1.6. Tukey's HSD Post Hoc Test.
Repellency comparison between oleic acid concentrations
Meanwhile, neither of the two factors
elicited a significant effect on the repellent
activity of Clove oil as shown in Table 1.7 (sex:
p=1.000 ; food: p=1.000). The results of the
negative control, however, showed that both
factors have a significant effect on repellency of
acetone (sex: p=0.035 ; food: p=0.035).
Specifically, Tukey's HSD test reveals that there
is a significant difference between male group
with food and female group without food
(p=0.023). However, no interaction between the
two factors was recorded to have a significant
effect as shown in Table 1.8.
Table 1.7. Clove Oil: Sex-Food factor
Table 1.8. Acetone: Sex-Food factor
Weight Change as factor
The current study also aimed at tracing
a correlation between the repellency of oleic acid
solutions and the changes in weight of the
subjects – whether gaining or losing weight
affects the repellency. The weight of the
cockroaches from each group were averaged
before and after the treatment. The changes in
weight were categorized to either "gained"
(positive difference in average weight) or "loss"
(negative difference in average weight). Through
One-way ANOVA, the significant difference in
terms of repellency between groups within each
set (30, 60, 100%) were plotted. Results show
that weight changes did not significantly affect
the repellent activity of both the 30% and 100%
oleic acid p=0.735 and p=0.571 respectively.
The 60% group, however, showed a significant
difference in repellency between groups that
gained weight and groups that lost weight
(p=0.002). In the 60% set, lower repellency was
traced from groups that experienced weight loss
which may indicate that the subjects from this
group became attracted to the solution.
Table 2.1. One-way ANOVA.
30% Oleic acid group: Weight change as factor
8
Table 2.2. One-way ANOVA Tukey HSD Test.
60% Oleic acid group: Weight change as factor
(*p<0.05, **p<0.01, ***p<0.001)
Table 2.3. One-way ANOVA.
100% Oleic acid group: Weight change as factor
Time as factor
To test whether the effects of the
produced solutions change with time, the results
from each concentration were tested using a
Two-way ANOVA. Results show that there is no
significant difference between all groups treated
with the oleic acid solutions in terms of
repellency on each mark time (p=0.717) (Figure
1.1). This indicates that the general repellency of
the oleic acid solutions did not significantly
change during the whole 10 hour exposure.
Figure 1. Repellency of oleic acid solutions over the 10 hour
exposure
Levels of Repellency
The current study utilized an
experiment container design with partitions
referred to as levels. These levels were used to
measure how far from treatment the subjects
were located. After a Two-way Analysis of
Variance, results show that there is a significant
difference in the number of feeding chamber and
repel chamber visits per level between both
control and experimental groups (p<0.001). As
shown in Figure 2, the most frequently visited
area in all groups except the 60% and negative
control was the FC followed by the visits on each
level which vary with the test chemicals. This
location is 40 cm away from the treatment
substance. On the other hand, the 5th level of the
repel chamber (95 cm away from treatment) was
the most visited area in both 60% and negative
control groups indicating a long range effect of
these chemicals.
The least visited area in all groups aside
from the positive control was the third level (75
cm away from treatment) of the repel chamber.
Moreover, the results from the acetone group
show an equal number of visits in Levels 1, 2
and 3. Level 1 and 3 had the same number of
visits for the 30% oleic acid group. While, the
same number of visits to Levels 2 and 3 was
observed from the positive control group.
9
Figure 2. Frequency of visits to RC levels and
FC
System Usability Survey
The results from the analysis of the
responses showed low rating equivalents. A
grade of F corresponding to "poor" was recorded
for two factors: Production/Market Cost and
Intended Purpose. While, both
Environmental-User Friendliness and Material
factors got a grade of D which corresponds to
"awful". Observation-wise, these low grades
were due to the counterbalancing of X and Y
values. The survey generally had high values for
X, however, also had corresponding high values
of Y. Thus, the system was rendered to have a
poor rating.
DISCUSSIONS
Positive Control
The repellency (%) of the positive
control was the highest among all the test
chemicals. The positive control testing was
repeated twice in order to validate if the system
was working (experiment room and container).
In the first trial (not included in paper), 10 ml of
100% Clove oil was smeared to a filter paper and
followed the same process of treatment. This
quantity was later changed due to overflowing of
the chemical on the petri dish. Further, the first
trial of the positive control resulted in high
repellency with only 5 Treatment chamber visits
recorded (=91.67% repellency). Despite such
results, the experiment was reconducted as some
conditions such as the darkening mechanism
were not consistent and even. On the second
trial, however, only 10 ml of Clove oil was left
available for the experiment. To address this
scarcity, 8 ml of Clove oil was mixed with 12 ml
of acetone producing a solution with 40% Clove
oil concentration and a total volume of 20 ml.
Each group was treated with 5 ml of the solution.
The results of which is what is used as the data
for Clove oil repellency (=96.67% repellency) as
the system was rendered working. Therefore, the
results from the positive control group support
the data gathered by (Omara et al., 2013) whose
findings also show that Clove oil has a high
repellent activity against American cockroaches
even at low concentrations (10%).
Oleic Acid
Meanwhile, among the varying oleic
acid solutions, the concentration that resulted in
the highest repellency (%) was the 30% oleic
acid followed by 60% and 100% oleic acid
respectively. Moreover, only the 60% group
showed a significant effect attributed by both sex
and food factors. Despite inducing a repellent
effect, the findings from the 30% and 60%
groups might not be as conclusive since the
carrier, which is acetone, also elicited a high
repellency even exceeding the repellent activity
of 60% and 100% oleic acid. This may explain
the negative correlation between repellency and
the concentration of the oleic acid solutions since
the acetone was also assumed to be acting on the
subjects; Lower oleic acid concentration
corresponds to a higher acetone concentration.
10
Nonetheless, a higher percentage repellency was
recorded from the 30% oleic acid group
compared to the negative control. This may
indicate either an underlying synergy between
the two chemicals or just the carrier
overwhelming the effects of the supposed active.
In the sense of synergy, since more acetone was
mixed in the 30% oleic acid, its elicited higher
repellency may be due to sufficient amount of
acetone to totally disperse all the oleic acid
components of the solution; The large amount of
the volatile carrier dissolved and perhaps
evaporated decent amount of oleic acid allowing
it to disperse more of the fatty acid and reach the
subjects efficiently. Otherwise, the case of
repellency may just have been due to the direct
effects of acetone.
Eitherway, an argument may arise as to
why the pure oleic acid induced an attracted or
rather "unaffected" behavior on the subjects. The
current study, unlike other existing similar
research, used a different approach on treating
the subjects with the test chemicals. In
comparison, most research testing chemical
repellency uses a set up that divides a filter paper
into halves and then treating one side with the
experimental solution and the other with either
the negative control or nothing. Here, the filter
paper is directly in contact with the experiment
container floor such that when the filter paper is
treated, the cockroaches may step on the treated
paper unless they refuse to do so – repellency.
This approach known as Filter-paper choice
assay (Chang et al., 2017) was used by Sittichok
et al. (2013), and Takawirapat et al. (2021) in
testing the repellency of plant essential oils
against American and German cockroaches. The
current research, however, was concerned with
producing a repellent. Such products rely heavily
on the fumigant effects of the active ingredient
rather than its toxicity upon contact with the
target species. However, no fumigant toxicity –
by which most repellents operate – was yet
recorded in the past literature about oleic acid
which may be due to the fact that oleic acid is
not as volatile as other studied organic repellents
such as essential oils. Instead, a finding suggests
that oleic acid has a high contact toxicity against
honeybees requiring only 1μl to be treated to the
species' abdomen to induce death (McAfee et al.,
2018). This notion was tested by the current
study as part of an external experiment (a
random test hence no data is collected other than
observations). Upon treating approximately 5 ml
of pure oleic acid to a plastic container (22 x 14
x 9 cm) holding 9 cockroaches of random sex, an
almost instantaneous death was observed once
the subjects' body made contact with the
substance which occupied the floor of the
container. After roughly 5 minutes, 100%
mortality was observed (9 out of 9 died). In
another random test, a newly caught male
cockroach was trapped to the ground using a
circular plastic canister. To test the contact
toxicity of the oleic acid, roughly 1 ml of the
substance was pipetted to an ordinary bond paper
and was slided under the canister such that the
cockroach stepped on it. Almost immediately,
the trapped subject avoided the treated portion of
the paper and persisted to do so for the following
minutes. One member tried to keep its contact
with the treated paper by gently moving the
canister and the paper to match the location of
the trapped cockroach. Eventually, the cockroach
showed signs of weakness and slow movements.
After some more exposure time, the cockroach
was found dead on the spot where the oleic acid
was treated. These two experiments, although no
data was collected as they were completely
random, demonstrated the high toxicity of oleic
acid on American cockroaches similar to its
effect on honeybees. In line with this, Baldwin et
al. (2008) shows that potassium oleate, a
potassium salt of oleic acid, elicited the highest
contact toxicity among all the fatty acid salts
tested in the study on American cockroaches.
These two findings may prove the toxicity of
oleic acid to American cockroaches upon contact
making the substance plausible for cockroach
control. However, the current study used a
fumigant testing approach by treating the
solutions to a filter paper fitted into a petri dish
and allowing it to reach the olfactory senses of
the subjects, thus, there was no direct physical
interaction between the cockroaches and the
oleic acid solutions. This may explain the
"unaffected" behavior of the subjects towards the
100% oleic acid since they were only exposed to
its presence by air which is currently not proven
to be effective.
Negative Control
On the other hand, the results from the
negative control of the current study oppose the
recurring conclusion in multiple existing
research which mostly shows that acetone has no
significant repellent activity against various
species of cockroaches including the American
cockroach. Acetone is widely acknowledged and
used as a carrier for testing organic cockroach
repellents. For instance, multiple research on the
11
repellency of different essential oils on American
cockroaches utilizes acetone as its carrier. In a
study on the repellency of nine essential oil
constituents conducted by Ngoh et al. (1998),
acetone did not show significant repellency when
used as the control substance. Meanwhile,
another study by Appel et al. (2001), shows that
acetone induced a low repellent effect on
American cockroaches over the 14 day
experiment. Similarly, Schultz et al. (2004)
showed that acetone as control demonstrated a
mean repellency of 50% which may indicate that
there may be cases of repellency, however, was
concluded by the study to be statistically
insignificant. Moreover, Omara et al. (2013)
posits a similar result in a choice test where
acetone was treated on one side of the container
and the experimental substance on the other side.
Results show that the subjects were repelled by
the experimental substance rather than by the
acetone indicating less repulsion from the
control. It is also noteworthy that this acetone
dilution approach is utilized in studies involving
other species of cockroaches with results similar
to that of the aforementioned: Liu et al. (2011)
on German cockroaches; and Sharififard et al.
(2016) on Brown-banded cockroaches to name a
few.
Multiple research also suggests that
acetone induces a low to zero chance of
mortality in American and other species of
cockroaches (Kaakeh et al., 1997; Babri et al.,
2012; Sharawi et al. 2013; Yeom et al., 2013).
The cited studies seem to oppose the findings of
the current research. However, a similar case was
found in a study conducted by Ubulom et al.
(2021) which also used acetone as a control
substance but of lower purity (20%
concentration). The study showed that the
repellent activity of acetone exceeded that of the
essential oil that was being tested. Thus, the
reason for the recorded repellency of the acetone
used in the current study might be due to its
uncertain purity which, although validated by the
seller to be pure acetone, was not indicated in the
product label itself. As such, the research claims
that there was a gap with the materials used
specifically in the brand and perhaps the purity
of the acetone, and other factors which could
have been executed efficiently if held in
laboratories.
Context Dependent Effect of Oleic Acid
Moreover, no conclusion can be made
on the repellency of the oleic acid solutions,
specifically the 30% group despite having a
higher repellency percentage compared to pure
acetone since the 100% oleic acid group showed
low levels of repellency. In line with the latter
statement, the researchers suggest that even
though oleic acid is proven to be one of the most
prevalent chemicals released by cockroaches
upon death, it may not be enough to induce a
repellent effect; Other chemicals are synergizing
with this fatty acid which is what may be causing
the repulsion from the source of this so-called
necromone. This may be the case for the results
that Rollo et al. (1995) reported since they
included dead intact cockroaches in their
experiment, which means oleic acid may not be
the only chemical affecting the behavior of the
subjects.
CONCLUSION
According to the data collected in the
current study, the brand of acetone used does not
appear to be a feasible option as the carrier of
oleic acid. Nonetheless, despite high contact
toxicity, pure oleic acid showed low levels of
repellency rooted to its low volatility.
Additionally, a short ranged effect was observed
from the oleic acid solutions as the most
frequently visited area was the Feeding chamber
(40 cm from treatment) aside from the 60%
group which, despite lower repellency than the
30% group, had Level 5 as the most visited area
of the Repel chamber. Moreover, no conclusive
data showed that sex, food factor and even
weight change had a significant effect on the
repellent activity of oleic acid, hence, no
selective repellency was recorded.
RECOMMENDATIONS
The current research has definite
limitations such as in the chemical carrier used,
and the conditions including the experiment
room set up, experiment container, light
mechanism, temperature, and humidity which
was not as consistent as what laboratory settings
and secured chemical sources can offer.
Container-wise, a clear material like glass would
have been preferable over the used alternative
which was made of wood and plastic instead.
Moreover, the use of a larger subject population
may help in producing more accurate and
statistically diverse results. In short, a
replicability test of the current study must be
12
done in order to achieve empirical conclusions
on the topic and methods used. Finally, since
oleic acid works more efficiently on cockroaches
upon contact, a product that utilizes this
mechanism might be feasible such as a repellent
in a form of chalk similar to those available in
the market.
ACKNOWLEDGEMENT
The researchers express their utmost
gratitude to their research professor, Ms. Alora P.
Pozon, for actively guiding and suggesting
meaningful ideas that were helpful for the
completion of this research. The researchers also
acknowledge the sense of focus that Mr. Lyndon
Mercado instilled in the members by having
them write journal reports to occasionally record
and recall the progress of the study.
The researchers would also like to give
special thanks to the family of the researchers.
To one of the group member's mother, Mrs. Sta
Ana, for having provided the fund to purchase
the Oleic acid used in the research. To the
parents of another member, Ms. Josephine
Medrana and her family, for partially funding the
research and allowing the experiments to be
conducted in the vicinity of their house. To Mr.
Modesto Medrana and Mrs. Fe Medrana for
giving the permission to use their family wifi
during the research process.
The researchers also recognize the
deliberate effort and time granted by the students
of the STEM 12 Department for participating in
the system usability survey. Their coordination
was of vital part for the research completion.
Additionally, the analysis of the survey would
not have been possible without the help of a
fellow student, Ms. Fatima Ll. Asanza, who used
her laptop in the numerical interpretation of the
survey responses.
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16