Journal of Entomology and Zoology Studies 2018; 6(4): 185-189

E-ISSN: 2320-7078
P-ISSN: 2349-6800
JEZS 2018; 6(4): 185-189
Preparation of dust formulation of essential and
© 2018 JEZS
Received: 29-05-2018
aromatic oils and testing the bioefficacy against
Accepted: 30-06-2018 pulse beetle Callosobruchus maculatus (Fab.)
Manju K
M.Sc. (Agri.) Entomology,
(Coleoptera: Bruchidae) in green gram storage
Department of Agricultural
Entomology, TNAU Agricultural
College and Research Institute, Manju K, J Jayaraj and M Shanthi
Madurai, Tamil Nadu, India
Abstract
J Jayaraj
In the present study, the toxicity of seven essential and aromatic oils viz; neem oil, mint oil, citronella oil,
Department of Agricultural
Entomology, TNAU Agricultural
mustard oil, geranium oil, coconut oil and eucalyptus oil were evaluated against pulse beetle,
College and Research Institute, Callosobruchus maculatus Fab. in green gram seeds through contact, fumigant, repellency and settling
Madurai, Tamil Nadu, India behavior bioassay. All the treatments were found significantly superior in causing mortality of the pulse
beetle compared to the untreated check. Citronella oil showed a maximum mortality for both fumigant
M Shanthi and contact toxicity assays (100% and 93.75%) and strongest repellent (100%) against pulse beetle. The
Department of Agricultural toxicity of the oils is in the order of efficacy is citronella ˃ geranium ˃ mint ˃ mustard ˃ coconut ˃ neem
Entomology, TNAU Agricultural ˃ eucalyptus. Generally the oil persistency was minimum; hence in order to increase the persistency, the
College and Research Institute, dust formulation of essential oils of citronella, geranium and mint oil were prepared at 5, 10 and 15
Madurai, Tamil Nadu, India percent. The results indicated, the dust formulations of citronella and mint oil were found to be effective
at 10% and geranium oil at 15%.

Keywords: green gram, Callosobruchus maculatus, essential oils, aromatic oils

1. Introduction
Pulses are the major source of protein in the developing world [3]. Green gram is the most
important pulse crop. The 68th UN General Assembly declared, 2016 as the International Year
of Pulses, which encouraging awareness on the nutritional composition of green gram as
follows: 100g of green gram provides 30 calories and consist of approximately 3g proteins, 6g
carbohydrates and 2g dietary fibers [3]. Green gram seeds are bound to show rapid and great
losses both qualitatively and quantitatively due to the attack of bruchid insects at post-harvest
stages [12]. Among the bruchids C. maculatus can cause damage of pulse seeds upto cent
percent during storage [5]. Protecting stored pulses against C. maculatus is known to depend on
the use of synthetic chemicals. Nowadays broad spectrum insecticids have been reported to
cause development of resistance in insect populations [4].
Some products extracted from aromatic plants shows insecticidal activity against bruichid
species [10] Essential and aromatic oils can be used as fumigant, contact insecticides, and as
repellent in the control of stored grain pests [7, 11]. Plant material based essential oils are target
specific, non-toxic to human and beneficial organisms, less prone to insect resistance and
resurgence and biodegradable and also act as promising grain protectants [18]. In light of the
adverse effect of insecticides on the environment, these methods of pest control are now
attracting greater attention and research input [19]. Hence the present study was conducted to
investigate the potential of toxicity and repellent action of selected essential and aromatic oils
on pulse beetle C. maculatus in stored green gram.

2. Materials and methods

The present study was conducted in the Insectary, Department of Agricultural Entomology,
Correspondence Agricultural College and Research Institute, Madurai, Tamil Nadu during 2017 – 2018.
Manju K
M.Sc. (Agri.) Entomology,
Department of Agricultural 2.1 Mass culturing of pulse beetle, C. maculatus
Entomology, TNAU Agricultural Pulse beetles required for the study were mass reared on green gram (CO- 5) in the transparent
College and Research Institute, plastic containers of 600 ml at Insectary. Each container was filled with 200g of green gram
Madurai, Tamil Nadu, India and ten pairs of matured adult beetles were released. The plastic jars were covered with muslin
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cloth tightly, secured with a rubber band. Provide air passage treated halves to untreated halves of the same dimensions
to avoid fungal growth and placed in dark to facilitate with cello tape. Ten newly emerged adults were released at
maximum oviposition maintained at a room temperature of 30 the centre of each petridish and subsequently covered. The
± 5°C and 70 ± 5% RH throughout the period of study [15]. seven treatments were replicated for 3 times. Insects that
Oviposition was completed within five days. After 25 to 30 settled on each half of the filter paper disc were counted after
days, adults that emerged from the culture were utilized for a time lapse of 1, 2 and 3 hour. The data were expressed as
maintenance of sub cultures following the same procedure as percent repellency [2]. The percentage repellency using the
described above. Freshly emerged adults were used for following formula.
conducting the experiments.

2.2 Essential and aromatic oils treatment

Neem oil, eucalyptus oil, citronella oil, geranium oil, coconut
oil, mustard oil and mint oil which had insecticidal and
repellency action were identified for this study. The oils were
made to the following concentration by using acetone as a Where,
solvent. A = Average number of insect present on untreated portion
B = Average number of insect present on the treated portion
Treatment Common name Scientific name Dose
T1 Neem oil Azadirachta indica 1% Repellency rate Class
T2 Eucalyptus oil Eucalyptus globules 0.5% >0.01-0.1% 0
T3 Citronella oil Cymbopogon nardus 1% 0.1-20% I
T4 Geranium oil Pelargonium graveolens 1% 20.1-40% II
T5 Coconut oil Cocos nucifera 1% 40.1-60% III
T6 Mustard oil Brassica juncea 1% 60.1-80% IV
T7 Mint oil Mentha longifolia 1% 80.1-100% V

2.3 Contact toxicity 2.6 Studies on settling behavior

The insecticidal activity of essential and aromatic oils against The one per cent oil solutions were sprayed outside the stored
C. maculatus was studied by direct contact application of bags (15×10cm) kept in the experimental cages. Fifty newly
various treatments of essential oils on filter paper [14]. The emerged adults were released in each cage (25×25×35cm).
prepared solvents were uniformly sprayed in the filter paper The seven treatments were sprayed in each cage using hand
and were allowed to evaporate for 20 min. The filter paper sprayer. The treatments were replicated for three times.
was then placed in the petriplate and followed by the release Untreated check was also maintained simultaneously. The
of three day old 20 numbers of unsexed adults separately into seven treatments were replicated for three times. Adult beetles
each petriplate. Each treatment had four replications. Insect that settled on each bags were counted 48 and 72 hour after
mortality was recorded after 24h exposure. the release.

2.7 Dust (D) formulation of essential oils

Dusts were made up of a finely ground mixture of low
concentration of active ingredient (10% or less by weight)
combined with very fine and dry inert carrier made from talc
in powder form [9]. The dust formulation of essential oils was
Where, prepared at 5% (0.52 ml/10g), 10% (1ml/10g) and 15%
X = Per cent living in the check (1.7ml/10g) and the dust was applied along with green gram
Y = Per cent living in the treatment seeds in plastic containers. For testing the persistency, the
X – Y = Per cent killed by the treatment dust formulation was applied immediately after preparation
and which was compared with the same formulation after one
2.4 Fumigant toxicity month. Released 12 pairs of 3 days old unsexed pulse beetle
Filter paper disc of 4 cm diameter were impregnated with adults and recorded the percentage mortality after 24hr. Three
different solvents of essential oils, to give equivalent fumigant replications of treatments and untreated checks were
concentration of 26.5µl/ 250ml in air [8]. The impregnated maintained.
filter paper was then attached to the inner surface of the screw
cap of the glass bottle (250ml). Twenty adults of known age 3. Results and Discussion
were released in each glass bottles before the cap were Considering the public demand for wholesome and pest free
screwed tightly. Number of dead insects was determined at 3, food products, there is a need for developing the biorational
6, 12, and 24 h after exposure [14] and the per cent mortality pest management strategies by reducing the use of contact
was recorded. chemical insecticides and fumigants in stored products.

2.5 Repellency test 3.1 Contact and fumigant toxicity

Filter papers (Whatman No.40) of 9 cm in diameter were cut The results clearly demonstrated that the selected essential
in to two halves. One ml of 10 percent solution prepared in and aromatic oils showed potent fumigant and contact activity
acetone and applied to one half of the filter paper (treated against the pulse beetle, C. maculatus. All the treatments were
half) and on the other half, 1 ml of acetone was applied as found significantly superior in mortality of the pulse beetle.
uniformly as possible with a pipette. The treated filter paper Among the contact toxicity assay, citronella oil 1% and mint
was then air dried. Full discs were then re-made by attaching oil 1% showed cent percent mortality with in 24hr (Table 1)
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followed by geranium oil 1% (96%); coconut oil 1% (35%), Dust formulation of 10 and 15% Citronella oil recorded cent
eucalyptus oil 0.5% (20%) and neem oil 1% (11.25%). The percent mortality on the day of preparation of dust
lowest mean mortality was in mustard oil 1% (8.75%). formulation and also 30 days after preparation the 15% dust
Among the fumigant toxicity assay, highest mortality percent formulation of mint oil effected 100% mortality on the day of
(93.75%) was showed by citronella oil after 24 hours (Table preparation of the dust and its effects reduce slightly after a
1). This was on par with geranium oil (87.5%), mint oil period of 30 days thus effecting only 97.2% mortality of C.
(79.5%) and eucalyptus oil (79.5%) followed by coconut oil maculatus in 24hr (Table 4). geranium 15% dust was effected
(16.37%) mustard oil (9.65%) and neem oil (8.05%). only 63.8% and 51.3% mortality after 1 and 30 day after
preparation of the formulation.
3.2 Repellency assay Increasing the concentration of the tested oil increased the
Among the essential oils tested citronella oil 1% (100%) rate of insect mortality. Higher concentrations (15%) of
(Class V) was the strongest repellent against C. maculatus by geranium, mint and citronella oils induced the highest
registering cent percent repellency (Class V) (Table 2). mortality rate against C. maculatus (57.5%, 98.6%, and
Followed by geranium oil 1% (97.7%) (Class IV) which was 100%), respectively. On the other hand, the lower
on par with citronella oil. The other oils significant repellent concentrations of 5 and 10 percent of geranium, mint and
activity with neem oil 1% (57.7%) (Class III) and followed by citronella oils induced a less mortality against C. maculatus
coconut oil (51.1%) (Class III) on par with each other. The (22.85%, 36.8%, 81.8%, 22.8%, 93% and 100%) respectively.
lowest repellency was recorded with the eucalyptus oil 0.5% The effect of dust formulation was higher when the dust
(20) (Class II). Similar results were also reported by [16], when applied immediately after preparation when compared with
essential oil and botanicals tested against pulse beetle, C. one month. Dust formulation of geranium oil showed the
maculatus in green gram, citronella oil showed highest higher mortality rate (27.7%, 29.03% and 63.8%) of C.
repellency (100%), toxicity effects, reduced oviposition and maculatus when compared with the dust formulation after one
no adult emergence (0.0%) at 14,15,16 and 17 days after month (18%, 16.6% and 51.3%) respectively. Similarly mint
treatment. oil had the highest mortality rate (37.5%, 95.8% and 100%) of
C. maculatus when compared with the dust formulation
3.3 Settling behavior prepared after one month (36.1%, 90.2% and 97.2%)
After four days pulse beetle showed no settling behavior (0%) respectively. [6] Also reported that citronella at 2.5 ml/kg of
to citronella oil 1% (Table 3) which was on par with geranium seed effectively controlled C. maculatus population by
oil (2%). followed by mustard oil (6.1%), mint oil (9.1%) reducing oviposition rate and these treatments also recorded
neem oil (13.2%) and coconut oil (21.3%). The high settling the least seed damage and weight loss due to pulse beetle
behavior was shown by eucalyptus oil with 76%. [17] Have infestation, as well as the highest percentage of gram seed
also proven that the tested essential and aromatic oils showed germination.
minimum settling behavior, oviposition and seed damage
(7.7%) when green gram was treated with citronella oil at five 4. Conclusion
per cent when compared with untreated check (27.35%). The results clearly showed that the tested essential and
aromatic oils in this study have affected the settling,
3.4 Dust (D) formulation of essential and aromatic oils oviposition and development of the pulse beetle. Thus they
Among the essential and aromatic oils, geranium, citronella have high value for utility as the most promising essential and
and mint oil showed promising result in repellency, contact aromatic oils for the control of pulse beetle damage in stored
and fumigant toxicity and settling behavior assays. The green gram. Among all the essential and aromatic oil
effective essential and aromatic oils namely geranium, treatments, dust formulation of citronella oil at 10 per cent
citronella and mint oil were prepared as dust formulations and can be recommended for the management of the pulse beetle
tested for repellent effect settling behavior of C. maculatus. in storage.

Table 1: Insecticidal activity of essential oils against Callosobruchus spp. in contact and fumigant toxicity assay.
Treatment Dose Contact toxicity mg/cm2 Fumigant toxicity mg/L
T1- Neem oil 1% 11.25 (19.4)d 8.05 (13.5)b
T2- Eucalyptus oil 0.5% 20 (26.0)d 78.75 (70.1)a
T3- Citronella oil 1% 100 (89.5)a 93.75 (82.1)a
T4- Geranium oil 1% 96 (81.8)b 87.5 (78.3)a
T5- Coconut oil 1% 35 (36.1)c 16.37 (12.6)b
T6- Mustard oil 1% 8.75 (16.7)d 9.65 (12.3)b
T7- Mint oil 1% 100 (89.5)a 79.5 (70.2)a
SEd 3.5182 14.4
CD(.05) 7.3164 30.1
CV 9.68 42.20
Letters followed by mean values are significantly different from each other by LSD. Values are
average of four replications. Original data were corrected by Abbott’s formula and then
transformed into arcsine percentage values during ANOVA test.

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Table 2: Average repellency of essential oils to Pulse beetle adults using treated filter paper assay
Average % Repellency rate at hours after insect
Mean repellency
Treatments release Repellency class
(%)
1 2 3
T1- Neem oil @ 1% 60 (50.7) 53.3 (46.8) 60 (50.7) 57.7 (49.4) III
T2- Eucalyptus oil @ 0.5% 26.7 (31.1) 13.3 (21.3) 20 (26.5) 20 (26.3) I
T3- Citronella oil @ 1% 100 (89.5) 100 (89.5) 100 (89.5) 100 (89.5) V
T4- Geranium oil @ 1% 86.7 (68.6) 73.3 (58.8) 66.67 (54.7) 97.7 (60.7) IV
T5- Coconut oil @ 1% 46.6 (43.0) 53.3 (46.8) 53.3 (46.8) 51.1 (45.5) III
T6- Mustard oil @ 1% 40 (39.2) 46.6 (43.0) 33.3 (35.2) 39.9 (39.1) II
T7- Mint oil @ 1% 53.3 (46.8) 40 (39.2) 33.3 (35.2) 30.2 (40.4) II
SEd 9.1873 10.3603 10.3603 3.9273
CD(.05) 19.4765 21.9631 21.9631 8.3255
CV% 26.8352 23.07 31.11 10.11
The average values were classified based on the rate of repellency (%) as given below; 0= ˃0.01 - ˂0.1, I= 0.1 - 20, II= 20.1 – 40.0,
III= 40.1 – 60.0, IV= 60.1 – 80.0, V= 80.1 – 100.0. Original data were corrected by Abbott’s formula and then transformed into arcsine
percentage values during ANOVA test.

Table 3: Mean settling behavior of Pulse beetle adults on green gram stored bags treated with various essential oils
Treatment Dose Mean settling behavior
T1- Neem oil 1% 13.2 (21.2)b
T2- Eucalyptus oil 0.5% 76 (60.6)d
T3- Citronella oil 1% 0 (0.4)a
T4- Geranium oil 1% 2 (8.1)a
T5- Coconut oil 1% 21.3 (27.4)c
T6- Mustard oil 1% 6.1 (14.0)a
T7- Mint oil 1% 9.1 (17.5)b
SEd 2.8390
CD(.05) 6.7135
CV % 15.56
Values are average of thee replications. Original data were corrected by Abbott’s
formula and transformed into arcsine percentage values during ANOVA test. Letters
followed by mean values are significantly different from each other by LSD.

Table 4: Efficacy of the dust formulation of selected essential oils

Dose Percent mortality rate at 24hr after insect release
Treatment Percent mean mortality
(%) Immediately after preparation of dust 1 month after preparation of dust
5 27.7 (31.7)e 18 (25.1)e 22.85 (28.4)e
Geranium oil 10 29.03 (32.5)e 16.6 (24.0)e 22.8 (28.2)e
15 63.8 (53.0)d 51.3 (45.7)d 57.5 (49.3)d
5 37.5 (37.7)e 36.1 (36.8)e 36.8 (37.2)e
10 95.8 (78.1)b 90.2 (71.7)b 93 (75.0)b
mint oil
15 100 (89.5)a 97.2 (80.3)a 98.6 (85.0)a
5 86.08 (68.2)c 77.6 (61.8)c 81.8 (65)c
10 100 (89.5)a 100 (89.5)a 100 (89.5)a
Citronella oil
15 100(89.5)a 100 (89.5)a 100 (89.5)a
SEd 2.7 4.8 4.3
CD(.05) 5.8 10.2 9.7
CV% 5.3 10.1 7.1
Values are average of thee replications. Transformed into arcsine percentage values during ANOVA test. Letters followed
by mean values are significantly different from each other by LSD.

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