Asian Journal of Research in Crop Science

Volume 9, Issue 4, Page 132-138, 2024; Article no.AJRCS.124954

ISSN: 2581-7167

Efficacy of Essential Oils Against Black

Bean Aphid (Aphis fabae Scopoli)
under Laboratory Conditions
Rabina Pandit a*, Bishnu Dawadi a, Amir Pandit a
and Gayatri Kumari Upadhaya b
a
Lamjung Campus, Institute of Agriculture and Animal Science, Tribhuvan University, Nepal.
b Campus of Live Sciences, Institute of Agriculture and Animal Science, Tribhuvan University, Nepal.

Authors’ contributions

This work was carried out in collaboration among all authors. All authors read and approved the final
manuscript.

Article Information
DOI: https://doi.org/10.9734/ajrcs/2024/v9i4305

Open Peer Review History:

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Received: 05/08/2024
Original Research Article Accepted: 11/10/2024
Published: 16/10/2024

ABSTRACT

The black bean aphid (Aphis fabae), poses a significant threat to cultivated crops, leading to food
insecurity and hunger concerns. Essential oils from plants offer promising low-risk alternatives to
synthetic chemicals for aphid control whose widespread use has raised serious environmental and
health issues. To assess the efficacy of plant essential oils, an experiment was carried out at the
Entomology Laboratory of Lamjung Campus, Tribhuvan University following a completely
randomized design with three replications using eight treatments including a control (distilled water
and ethanol). The essential oils used were Neem (Azadirachta indica), Eucalyptus (Eucalyptus
globulus), Palma rosa (Cymbopogon martini), Citronella (Cymbopogon winterianus), lemongrass
(Cymbopogon citratus), Juniper berry (Juniperus recurva), Mint (Mentha arvensis) and control

_____________________________________________________________________________________________________

*Corresponding author: E-mail: rabinapanditrp@gmail.com;

Cite as: Pandit, Rabina, Bishnu Dawadi, Amir Pandit, and Gayatri Kumari Upadhaya. 2024. “Efficacy of Essential Oils Against
Black Bean Aphid (Aphis Fabae Scopoli) under Laboratory Conditions”. Asian Journal of Research in Crop Science 9 (4):132-
38. https://doi.org/10.9734/ajrcs/2024/v9i4305.
 Pandit et al.; Asian J. Res. Crop Sci., vol. 9, no. 4, pp. 132-138, 2024; Article no.AJRCS.124954

(distilled water and ethanol) at the concentration of 1% and 2% with the exposure time of 12, 24 and
48 hours for contact mortality and 2 µL and 4 µL with the exposure time of 12 and 24 hours for
fumigation mortality. The result revealed that individual mortality rate increased with increasing oil
concentration and exposure time. Lemongrass with 2% concentration resulted in the high mortality
percentage (66.66%) whereas, Neem with 1% concentration demonstrated the lowest (23.3%) in
contact assay. Similarly, in fumigation assay, the highest mortality percentage was exhibited by 4 µL
Lemongrass (100%) when compared to 2 µL Neem and Juniper berry have lowest mortality
percentage (73.33%) after the control. Therefore, Lemongrass with 2% concentration for contact
assay and 4 µL Lemongrass for fumigation assay was considerably more effective than other used
essential oils. However, more research is necessary with locally available essential oils to assure the
long-term effectiveness in field and lab conditions with different concentrations and exposure time.
The result could benefit the farmer and gardener by developing essential oils as commercial
products to apply natural, eco-friendly and non-toxic management method which is alternative to
synthetic pesticides.

Keywords: Black bean aphid; essential oils; faba bean; mortality.

1. INTRODUCTION are complicated, organic, volatile compounds

with a distinct smell that are produced by
Aphids being the most hostile pests for almost all aromatic plants as secondary metabolites. They
cultivated crops, seriously diminish the are often produced via steam or hydro-
productivity of crops and cause spoilage by distillation, which Arabs invented in the Middle
sucking the sap from the leaves as well as Ages [11]. The majority of research
delicate stems [1]. The annual average yield concluded that essential oils have significant
losses brought on by aphids range from 30 to potential for pest management, particularly in
50% [2]. Aphid’s saliva includes non-enzymic, greenhouses and fields [12]. Essential oils are
reducing compounds, which, in the presence of very bioactive and can be very effective
oxidases, can mix with and inactivate protective insecticides. As a result, there should be a lot
phytochemicals, including those generated in more research done on them to help manage
response to damage and transmitted in the aphids [13].
phloem sieve tube sap on which aphid feed [3].
The black bean aphid (Aphis fabae) is a major 2. MATERIALS AND METHODS
pest of field beans which are sown in the season
of spring and the data of field studies have The experiment was carried out from March to
revealed that yield losses by the black bean April of 2023 in the entomology laboratory of the
aphid infestations can exceed 50% of the yield Institute of Agriculture and Animal Science,
[4]. Black bean aphids directly damage the plants Lamjung campus, which is in Sundarbazar, the
by feeding on the phloem that results in severe Lamjung district of Gandaki province. The site
reductions of plant development and productivity was located at an elevation of 610m above sea
[5]. In addition to direct plant injury, black bean level with latitude: 28.2847° N and longitude:
aphid infestation harms faba bean due to 84.3637° E under humid sub-tropical climate.
honeydew excretion on the leaves, which The highest and lowest temperatures recorded
interferes with physiological processes in the were, respectively, 28–30°C and 20–22°C.
host plant [6]. These aphids can indirectly Apterous (wingless) black bean aphid was
damage by acting as vectors for plant viruses [7] collected from the field of faba bean located
such as the bean leaf rolling virus and the faba nearby Lamjung campus by cutting aphid prone
bean necrotic yellows virus [8]. It has been long stem of plant with the help of sickle. The brushes
since botanical insecticides have been regarded were used to transfer the aphid from cut stem to
as acceptable alternatives for killing the harmful petri plate. The required essential oils were
insects because of their low retention in the obtained from Herbs Production and Processing
environment, minimal toxicity to mammals, better Company Limited, Kathmandu. Two factorial
selectivity and the popularity [9]. Numerous plant Completely Randomized Experimental Design
species' essential oils have a variety of was selected as the design of the research.
insecticidal, anti feedant, repellent, oviposition- Seven treatments of essential oils namely:
deterrent, growth-regulating, and anti vectorial Neem, Eucalyptus, Palma rosa, Citronella,
effects on nuisance insects [10]. Essential oils Lemongrass, Juniper berry, Mint along with one

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control were used. Three replication per using Duncan´s Multiple Range Test at 5% level
treatments were done. The aphid mortality was of significance.
assayed through two parameters during the
experiment. 3. RESULTS

2.1 Contact Mortality Assay (Leaf Dip 3.1 Effect of Treatments on the Contact
Bioassay) Mortality of Aphid

Using ethanol as a non-polar solvent, each At 12hrs, the highest mortality percentage was
treatment oil was dissolved before being utilized found in Lemongrass (26.66%) which was
to investigate the direct contact effect of essential statistically par with Palmarosa and Mint and the
oils on aphid species. There were two lowest mortality rate was in control .The lowest
concentration levels 1% and 2% -prepared for aphid mortality after control was in Neem (10%)
each treatment. In three replications, each which was statistically par with Citronella and
individual faba bean leaf was immersed in the oil Eucalyptus. Similarly, at 24hrs, highest aphid
solution of every treatment for about five mortality was seen in Lemongrass (43.33%) and
minutes. The volatile solvent was then allowed to lowest mortality was in Neem (15%) but
evaporate by letting them dry for a while. Eucalyptus and Juniper berry were also
Following treatment, the leaves were placed in significantly at par. In the 48hrs, similar result
sterile Petri plates with damp filter paper within. was observed where Lemongrass had the
In each petri dish, ten aphids were released and highest aphid mortality rate (61.66%) which was
given time to settle on the leaves that had been statistically similar with Palmarosa and
treated. Then the petri plate was covered by Eucalyptus. The lowest mortality percentage was
muslin cloth so that aphids present inside the shown by Eucalyptus (30%) which was
petri plate could not escaped away. After 12 statistically par with Neem.
hours and 24 hours, the number of dead aphids
was counted. Using a little brush to gently strike The mortality rate had increased with exposure
the aphid and keeping an eye out for any time and higher the concentration of Essential
movement of its legs or antennae, one may oils from 1% to 2%, more was the mortality rate.
identify dead aphids. From the interaction table, in 12hrs it was found
that Mint at 2% concentration revealed the
2.2 Fumigation Mortality Assay highest mortality percentage (33.33%) which was
statistically par with Lemongrass (30%) and
Saturated filter paper was stored in three layers Palmarosa (26.66%) but Neem at 1%
within sterile 9 cm petri dishes. At the bottom of concentration demonstrated lowest mortality
the filter sheets was a tiny, rectangular (5×2 cm) (6.66%). In 24hrs, Lemongrass with 2%
filter paper. Each essential oil was directly added concentration exhibited the highest mortality rate
to the rectangular piece of filter paper with a (53.33%) whereas lowest mortality was shown by
micropipette, at concentrations of 2 µL and 4 µL. Neem, juniper berry and citronella at 1%
To avoid direct contact between the oils and the concentration with 13.33% .Similarly, in 48hrs,
experimental pest, ten aphids were then Lemongrass with 2% mortality exhibited the
introduced onto the upper layer of the petri dish. highest mortality (66.66%) whereas Neem with
To keep the treated oils from leaking, parafilm 1% mortality showed the lowest (23.33%).
was then used to enclose the entire apparatus.
Similar methods were described in Chalise & 3.2 Effect of Treatments on Fumigation
Dawadi, 2019. Mortality of Aphid

Mortality of aphids was counted after 12hrs, From the table, at 12hrs, the highest mortality
24hrs and 48hrs. Mortality of treated aphid was percentage was shown by Palmarosa and Mint
then calculated by dividing the number of dead with 85% but statistically, it was similar with
aphids in petri plate by total number of aphids in Citronella, Eucalyptus and Lemongrass. Mortality
petri plate and expressing it in terms of percentage in Lemongrass was highest (98.33%)
percentage. The collected data of aphid mortality at 24hrs which were significantly par with
was processed with MS Excel 365 and analysis Citronella, Eucalyptus and Mint. Neem at 12hrs
of variance was done using R- Stat software (71.66%) and 24hrs (75%) showed the lowest
(version 4.1.0). Mean comparison was done mortality percentage after control.

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Table 1. Effect of Essential oils on aphid mortality on contact assay

Factor A Contact mortality after treatment application

(Essential oils)
12hrs 24hrs 48hrs
Neem 10.00c 15.00d 31.66cd
Eucalyptus 15.00bc 18.33cd 30.00d
Palmarosa 25.00a 35.00b 55.00a
Citronella 13.33bc 23.33c 43.33b
Lemongrass 26.66a 43.33a 61.66a
Juniper berry 16.66b 18.33cd 38.33bc
Mint 23.33a 33.33b 45.00b
Control 1.66d 6.66e 8.33e
CV% 17.73 13.13 15.19
F-test *** *** ***
LSD 5.36 6.57 6.99
SEM 2.97 4.28 5.82
Factor B 12hrs 24hrs 48hrs
(Concentration)
1% 12.08b 18.33b 32.91b
2% 20.83a 30.00a 45.41a
CV% 17.73 13.13 15.19
F-test *** *** ***
LSD 2.68 3.28 3.49
SEM 4.375 5.835 6.25
Grand mean 16.45 24.16 39.167
P value . ** **
*, ** and *** at P=.05, P=.01 and P=.001 level of significant respectively.

Table 2. Interaction of essential oils and concentration on aphid mortality in contact assay

Essential oils 12hrs 24hrs 48hrs

1% 2% 1% 2% 1% 2%
Neem 6.66ghi 13.33 efg 13.33gh 16.66fg 23.33g 40.00de
Eucalyptus 10.00fgh 20.00cde 10.00gh 26.66de 26.66fg 33.33ef
Palmarosa 23.33bcd 26.66abc 33.33cd 36.66bc 43.33cd 66.66a
Citronella 10.00fgh 16.66def 13.33gh 33.33cd 30.00fg 56.66b
Lemongrass 23.33bcd 30.00ab 33.33cd 53.33a 56.66b 66.66a
Juniper berry 10.00fgh 23.33bcd 13.33gh 23.33ef 33.33ef 43.33cd
Mint 13.33efg 33.33a 23.33ef 43.33b 40.00de 50.00bc
Control 0.00i 3.33hi 6.66h 6.66h 10.00h 6.66h
LSD 7.59 3.28 9.89
Grand mean 16.45 24.16 39.16
CV% 17.73 24.23 15.19

The mortality rate had increased with 4. DISCUSSION

exposure time and concentration of Essential oils
from 1 µl to 2 µl. From the interaction Tomova et al. [13], Ebrahimi et al. [9], Alghamdi
table, in 12hrs, it was observed that [2] also reported that the action of essential oils
Lemongrass with 4 µl concentration on the aphid is dosage dependent that the
showed the highest mortality (93.33%) and in individual mortality percentage increased with
24hrs, highest mortality was observed in increasing oil concentration. Hence, lemongrass
Lemongrass with 4 µl concentration was effective against black bean aphid in both
(100%) whereas lowest mortality was contact and fumigation assay which
shown by 2 µl Neem and Juniper berry with was in agreement with Shahid Nisar et al.
73.33%. [14] reported that the best results with the

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Table 3. Effect of Essential oils on aphid mortality in fumigation assay

Factor A (Essential oils) Fumigation mortality after using treatments

12hrs 24hrs
Neem 71.66c 75.00d
Eucalyptus 81.66 ab 95.00ab
Palmarosa 85.00 a 90.00bc
Citronella 83.33 ab 96.66ab
Lemongrass 78.33 abc 98.33a
Juniper berry 76.66 bc 83.33c
Mint 85.00 a 95.00ab
Control 5.00d 6.66e
CV% 8.39 7.21
F-test *** ***
LSD 6.99 6.78
SEM 9.54 10.83
Factor B 12hrs 24hrs
(concentration)
2µl 66.25b 77.08b
4 µl 75.41a 82.91a
CV% 8.39 7.21
F-test *** **
LSD 3.49 3.39
SEM 4.58 2.91
Grand mean 70.83 80.00
P value *** .
*, ** and *** at P=.05, P=.01 and P=.001 level of significant respectively

Table 4. Interaction of essential oils and concentration on aphid mortality in fumigation assay

Essential oils 12hrs 24hrs

2µL 4 µL 2 µL 4 µL
Neem 66.6ef 76.6cd 73.33d 76.66d
Eucalyptus 73.3de 90.0ab 93.3abc 96.6ab
Palmarosa 83.33 bc 86.6ab 86.66c 93.3abc
Citronella 83.33 bc 83.3bc 96.66ab 96.6ab
Lemongrass 63.33 f 93.33a 96.66ab 100.0a
Juniper berry 70.0 def 83.3bc 73.33d 93.3abc
Mint 83.33 bc 86.6ab 90.00bc 100.0a
Control 6.6g 3.3g 6.66e 6.66e
CV 8.39 7.21
LSD 7.29 9.59
Grand mean 70.83 7.21

highest mortality of aphid was shown by present in the oil of Lemongrass attributed to its
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high toxicity against Aphis citricola. The various imbalance, membrane damage, and cytotoxicity
bioactive cyclic and acyclic terpene constituents in the host [17]. In addition to its major

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