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Tithonia diversifolia (Hemsl.) A. Gray Essential Oil: A Potential
Biopesticide for Management of Sitophilus zeamais (Maize Weevils)
Emmanuel Amukohe Shikanga
Department of Chemistry, Maseno University, Private Bag Maseno, Kenya
DOI: https://doi.org/10.51584/IJRIAS.2025.1010000061
Received: 22 September 2025; Accepted: 28 September 2025; Published: 05 November 2025
ABSTRACT
This study evaluated the pesticidal activities of essential oils (EO) from the leaves of T. diversifolia against
Sitophilus zeamais using fumigation, repellency and contact toxicity assays. Forty-five compounds were
identified in the EO constituting 96.7% of the oil from GC/MS analysis. The main constituents included α-
pinene (42.2%), β-pinene (16.2%) and β-caryophyllene (12.2%). The oil displayed strong activities against
adult insects as expressed by fumigant activity with LC50 value 10.2 mg of oil/L of air (p<0.05), a contact
toxicity mortality with LD50 value of 12.3 μg/adult insect and class III repellency achieved within 30 min at a
very low conc (30 ul/cm2 paper discs). The high potency of the oils could be attributed to the major
components (α-pinene, β-pinene and β-caryophyllene), making it a potential biopesticide for protection of
maize grains from Sitophilus zeamais.
Keywords: Tithonia diversifolia, essential oils, Sitophilus zeamais, pesticidal.
INTRODUCTION
The repetitive use of synthetic pesticides for protection of cereal grains and control of pests may result in
development of resistant insect populations, toxic residues adhering to food and accumulation in the
environment [1]. The use of biopesticides including essential oils (EO) to control pests has gained momentum
since they are effective, present smaller effects on the fauna and flora, pose greater availability, and degrade
easily compared to synthetic compounds [2]. They are also cheap, non-toxic to non-target organism, and are
less likely to result in resistance in the target organisms. Many plants that are environmentally-friendly and
readily available in nature can be used in minimization of grain losses during storage by control of pests in
stored products. The susceptibility of crop plants to insect pests and the increase in costs of synthetic pesticides
have caused the need for application of alternative effective and biodegradable substances including EO,
extracts and isolated compounds [3].
Plant secondary metabolites including alkaloids, monoterpenes, sesquiterpenes, phenols and coumarins, are
known to exhibit toxicity, antifeedant, repellency and growth regulating effects against a range of insect pests
including Sitophilus zeamais (Motsch.) (Coleoptera: Curculionidae), commonly referred to as the maize weevil
[4]. This is a storage pest that has been associated with destruction of maize grains resulting in a decline in
production over the years [5].
Most synthetic pesticides are environmental hazards causing serious problems including direct toxicity to
predators, pollinators, fish and man. Over the years, synthetic pesticides such as organochlorines,
organophosphates, carbamates, pyrethroids and neonicotinoids have been effective for control of S. zeamais
[6]. However, the use of these chemicals has resulted in numerous challenges including development of
resistant insect strains, toxic residues in foods and humans, workers’ safety and most of them are expensive
[6]. Tithonia diversifolia [Hemsley] A. Gray (Asteraceae), also referred to as wild sunflower is a perennial
plant known to contain compounds with antioxidants, antibacterial, pesticidal and medicinal properties [7]. It
is commonly found scattered along rivers and roadsides, wastelands, hedges, along narrow paths and around
crop fields, and homesteads [8]. It grows both within and outside the tropics parts of central, south and North
America, Pacific islands like Hawaii, Guam, New Caledonia, French, Polynesia, Palau and Indian island [9]
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and in tropical parts of Asia as well as Africa [10]. In Africa, it is common in many countries including Egypt,
Guinea, Nigeria Cameroon, South Africa, Zimbabwe, Malawi, Congo, Ethiopia, Tanzania, Uganda and Kenya
[7, 11, 12]. In Kenya it was first introduced as an ornamental plant in 1940s and now has since spread to many
regions including Coastal, Mount Kenya, parts of the Rift Valley and western Kenya [7, 13, 19].
T. diversifolia has been evaluated for antibacterial, antimalarial and insecticidal activities. It has been used for
treatment of menstrual pains and diabetes mellitus and for control of insects pests such as aphids, beetles,
mosquitoes, maize weevil [13, 14]. Most studies have focused on the potency of extracts (aqueous, methanol,
ethyl acecate, DCM and hexane extracts) against insects such as aphids, beetles, mosquitoes, maize weevil,
bean weevil and whiteflies among others [7, 14, 15, 16]. However, information on activity of essential oils
against insect pests including Sitophilus zeamais is limited. This study aimed at characterizing of the EO of T.
diversifolia and determination of its pesticidal activities against S. zeamais.
MATERIALS AND METHODS
Rearing of Sitophilus zeamais
The insects were cultured under laboratory conditions of 25 ± 2 ⁰C and 65 ± 5% relative humidity in Maseno
University Department of Zoology laboratory. A mass of 300 g of the maize grains were transferred into 1 L
glass jars which were covered with a fine mesh cloth for ventilation and to prevent the insects from escaping.
Fifty adult of S. zeamais were then introduced into the each glass jar and kept in the incubated for seven days
for the insects to lay eggs and multiply.
Plant material, extraction and analysis of the essential oil
Fresh leaves of T. diversifolia were collected in April 2019 at the beginning of flowering season from Maseno
forest within Maseno University. The species were identified at Department of Botany Maseno University and
a voucher specimen (MU-TDV-020-2019) was deposited. 500 g of fresh leaves was subjected to hydro
distillation in a Clevenger-type apparatus for 6 h. The EO obtained was extracted with n-hexane, dried using
anhydrous sodium sulphate and analysed using gas chromatography mass spectrometry (GC/MS). The GC/MS
system used comprised of a HP 6890 Series GC system (Agilent Technology, California, USA), coupled with
a 5973 mass selective detector and an HP-5ms fused silica capillary column with a 5% phenyl-
methylpolysiloxane stationary phase (30 m X 0.32 mm X 0.25 μm). The oven temperature program was
initiated at 40 °C for 1 min then raised to 230 °C at a rate of 3 °C min-1 for 10 min. Helium was used as the
carrier gas at 1.0 mLmin-1 flow rate with a split ratio of 1/50. Detector and injector temperatures were 250 and
230 °C, respectively and spectra were obtained following electron impact ionization at 70 eV (35 to 550 m/z).
The essential oil compounds were identified by comparing their retention indices, mass spectra fragmentation
with those on the NIST® library version 2004.
Fumigant toxicity
Fumigation assay involved impregnating 3-cm diameter Whatman No. 41 filter paper discs with 10 μL of oil at
a different concentration (0: acetone, 1.6, 3.2, 4.6 and 6.15 mg/L in acetone). This corresponded to 0, 6.4, 12.8,
18.6 and 24.6 mg of oil/L of air in the glass jars. Acetone was used as negative control while MeBr was used
as the positive control. Each impregnated filter paper was allowed to dry for 15 min and thereafter, attached to
the bottom of the screw caps of a gas tight 250 mL glass jars. Preliminary experiments showed that that 15 s
were adequate for the evaporation of acetone. The caps were tightly screwed on glass jars containing 10 adult
insects. The insects had no contact with the suspended impregnated filter paper and stayed at the bottom of the
jars throughout the experiments. Control insects were kept under the same conditions although the filter paper
discs were impregnated with only acetone but no oil. The treatments and controls were incubated at 28–30 °C
and 70–80% relative humidity for 24 and 48 h. This was done in five replicates. The number of dead and live
insects in each jar was counted at the end of a 24 and 48 h exposure periods. Insects were considered dead
when no leg or antennal movements were observed. Percentage insect mortality was calculated using Abbott’s
[32] correction formula for natural mortality in untreated controls. Experiments were arranged in a completely
randomized design. The LC50 (conc required to attain 50% mortality for a given time for each conc of extract)
values with their fiducial limits were calculated by probit analysis using SPSS version 16.0 software package.
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Insect repellency assay
Insect repellency tests were conducted according to method by Islam et al. [33] with slight modifications. 14
cm diameter filter papers (Whatman no. 41) were each placed in a petri dish with same size. A volume of 1 ml
essential oil was then was uniformly applied to half of the papers to result in concentrations of 10, 20, 30, 40
and 50 ul/cm2 and absolute acetone was used a negative control was applied to the rest of the filter papers. The
contents of each petri dishe were left for 10 min at 25 ± 1 °C and 70 ± 10% relative humidity to evaporate the
solvent and ten S. zeamais adults were placed in the centre of each dish. The numbers of weevils in the control
(NC) and treatment (NT) dishes were recorded after 30, 60, and 90 min in 5 replicates. The percentage of
repellency (PR) was obtained by using the equation 1.
Repellency was further classified into class 0 (PR ≤ 0.1%), class I (PR = 0.1–20%), class II (PR = 20.1–40%),
class III (PR = 40.1–60%), class IV (PR = 60.1–80%), and class V (PR = 80.1–100%) in as described by Benzi
et al. [26].
Contact Toxicity
Determination of contact toxicity was conducted as described by [19] with modifications. Five serial dilutions
of EO (120.0, 200.0, 350.0, 500.0 mg/L) were prepared using acetone which was also used as negative control
while pyrethrine (120.0 mg/L) was used as positive control. An aliquot of 50 μL of each dilution and control
was applied topically to the dorsal thorax of each insect, using a 1 mL Hamilton micropipette. This translated
to conc levels of 6.0, 10.0, 17.5 and 25.0 µg of EO/adult insect respectively for the EO, and 0 and 6.0 µg/adult
insect for the negative and positive controls respectively. For each concentration level, ten insects were treated
and transferred to glass vials with culture media (maize flour). This was repeated for the insects treated with
the control solvent. The experiment was conducted in six replicates. The vials containing the insects (10
insects/vial) were kept in incubators at 29-30 °C and 70- 80% relative humidity. Mortality of the insects was
observed after 6, 12, 24, 36 and 48 h. The observed mortality data were corrected using Abbott’s [32]
correction formula. Results from all replicates were subjected to probit by analysis using SPSS version 16.0
software package for determination of LD50 (the amount of a chemical that is lethal to one-half (50%) of the
experimental animals exposed to it).
RESULTS AND DISCUSSION
Characterization of essential oil from T. diversifolia leaves
The essential oil (EO) obtained from hydro-distillation of T. diversifolia leaves was light yellow in colour with
a yield of 0.0025% w/w and the density of the concentrated essential oil was determined to be 0.80 g/mL. This
yield was higher than that reported by Wanzala et al. [13] from hydro distillation of T. diversifolia leaves
obtained from the slopes of Mt Elgon (0.000015% w/w) in western Kenya but lower than value reported by
Moronkola et al. [17] (0.019% w/w) from Nigeria. The difference in the yields could be due to changes in
environmental and climatic conditions in the different places [18]. The oil was observed to be insoluble in
water but soluble in organic solvents including n-hexane, diethyl ether, DCM and DMSO.
Forty five compounds were identified in the EO constituting 96.7% of the oil (Table 1). The major components
included α-pinene (42.2%), β-pinene (16.2%), β-caryophyllene (12.2%), followed by limonene (8.5%) and
(E)-nerolidol (6.5%). Although the percentage weight of monoterpenes (68.3% w/w) in the oil was higher than
those of sesquiterpenoids (29.3% w/w), a majority of the constituents comprised of sesquiterpenoids
representing 46.7% (21 of the 45 components), followed by monoterpenes accounting for 42.2% (19 of the 45
compounds) of the whole oil. The main components were similar to those obtained from leaves of T.
diversifolia obtained from Mt Elgon in western Kenya (α-pinene: 63.64%, β-pinene: 15.0%, iso-caryophyllene:
7.62% and nerolidol: 3.70%) [13], and in Osun state, Nigeria (α-pinene: 32.9%), β-pinene (10.9%), and β-
caryophyllene (20.8%) [17], although their respective percentages in the oils varied.
PR = x 100 …….. Equation 1
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Table 1: Composition of the essential oil obtained from the leaves of T. diversifolia
Peak No.
Compounds
RI
1
α-Pinene
942
2
Camphene
953
3
Sabinene
969
4
β-Pinene
980
5
β-Myrcene
990
6
α-Phellandrene
1002
7
∂-3-carene
1011
8
p-Cymene
1020
9
Limonene
1027
10
1, 8-Cineol
1030
11
(Z)-β-Ocimene
1032
12
(E)-β-Ocimene
1050
13
α –Terpinene
1054
14
α-Terpinolene
1088
15
Linalool
1100
16
Terpinen-4-ol
1177
17
α-Terpineol
1188
18
Decane-2,6,8-Trimethyl
1205
19
Geraniol
1249
20
α-Cubebene
1345
21
α-Copaene
1376
22
α-Gurjunene
1409
23
Bicyclo-2,2,2-octa-2,5-diene, 1,2,3,6-tetramethyl
1412
24
β-caryophyllene
1419
25
β-Gurjunene
1428
26
Aromadendrene
1440
27
α-Humulene
1452
28
β-Humulene
1454
29
(E)- β-Farnesene
1456
30
β-Ionane
1462
31
Germacrene D
1481
32
1-Tridecanone
1497
33
β-Bisabolene
1509
34
γ-Cadinene
1513
35
δ-Cadinene
1525
36
Elemicin
1555
37
(E)-Nerolidol
1564
38
Caryphyllene oxide
1581
39
Juniper camphor
1594
40
Cyclodecene
1605
41
Pentadecanone
1612
42
Cycloundecanone
1630
43
α-Bisabolol
1685
44
Farnesol
1717
45
1-Octadecanol
2024
Total
*Tr: trace, i.e., <0.1%; aRetention index on HP-5ms fused silica capillary column.
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Fumigant toxicity assay
High mortality rates (>80%) were obtained with concentrations T. deversifolia EO above 18.6 mg of oil/L of
air after 24 h and 12.8 mg of oil/L of air for 48 h exposure times (Table 2). Concentrations of 24.6 and 18.6 mg
of oil/L of air for 24 h and 48 h exposure times respectively, were adequate to result in 100% death of the
insects indicating a dose/time-dependent response in mortality rates of S. zeamais. The essential oil was less
toxic than the MeBr (positive control) which attained 100% mortality within 24 h at a conc of 12.8 mg of oil/L.
Table 2. Cumulative percentage mortality of adult S. zeamais after 24 and 48 h at different concentrations of T.
diversifolia essential oil by fumigation assay.
Conc of
EO(mg/L of air)
%mortality
24 h
%mortality
after 48 h
LC50 at 24 h
(mg/L of air)
Slope ± SE
Chi-
square
df
p
0(Acetone)
0.0±0.0a
0.0±0.0a
10.2
(8.7- 12.3)
1.21±0.18
1.98
3
0.54
6.4
32.5±3.5b
60.6±4.4b
12.8
60.4±5.1c
84.8±6.0c
18.6
81.1±6.4d
100.0±0.0d
24.6
100.0±0.e
100.0±0.0d
MeBr
100.0±0.0e
100.0±0.0d
*EO=essential oil. **Values on the table represent mean±SD (N= 5). ***Column means followed by different
letters are significantly different by Turkey test (p<0.05). Acetone solvent =-ve control; MeBr = +ve control.
The LC50 of the EO was 10.2 mg of oil/L of air at 95% upper and lower fiducial limits (FL) (8.7 to 12.3 mg
oil/L of air), for 24 h exposure time. The linear regression equation of S. zeamais mortality after 24 h duration
was y= 4.84x - 3.26. The activity of the essential oil was lower than that of methyl bromide (MeBr) which has
been used as the positive control in the study with a reported LC50 value of 0.67 mg oil/L air [19].
This EO exhibited stronger fumigant activity against S. zeamais than that obtained from of aerial parts of A.
subdigitata with LC50 of 17.01 mg oil/L [20], A. sieversiana with LC50 of 15.0 mg/L [21] and Illicium simonsii
with LC50 of 14.95 mg/L [22]. The EO of T. diversifolia had weaker fumigant activity compared to oil from
Artemisia giraldii with LC50 of 6.29 mg oil/L of air [20], Murraya exotica with LC50 of 8.29 mg/L of air [23]
and A. mongolica with LC50 of 7.35 mg/L of air [21]. Therefore, the essential oil of T. diversifolia has great
potential for used as a possible natural fumigant for the control of S. zeamais.
Insect repellency assay
The EO from T. diversifolia leaves repelled S. zeamais adults in a dose and time dependent manner as
expressed in Table 3. The essential oil attained class III repellency within 30 min for a 30 ul/cm2. Similar level
of repellency was attained at 60 min for essential oil conc of 20 ul/cm2 and 90 min for a conc of 10 ul/cm2.
This results are in agreement with results by Tavares et al. [24], where methanol extract of T. diversifolia
flowers exhibited class III of repellency after 90 min exposure against Sitophilus zeamais adults in corn grains.
Repellency against S. zeamais adults was also observed in a dose and time dependent manner for DCM and
ethylacetate extracts of T. diversifolia as indicated by Gitahi et al. [25]. The major components in the DCM
extract were β-Amyrin, squalene, and hexadecanoic acid, whereas those of the ethyl acetate extract were
hexadecanoic acid, squalene, methyl linoleat and phytol. These components differed from the major
components in the EO of T. diversifolia, indicating that repellency in the different case studies could be due to
different components.
Table 3. Repellency of Sitophilus zeamais adult insects by different concentrations of essential oil from the
leaves of T. diversifolia at different exposure times.
Conc of EO (ul/cm2)
% mean repellency of adult insects
30 min
60 min
90 min
Acetone (control)
0.0±0.0a (I)
0.0±0.0a (I)
0.0±0.0a (I)
10
5.0±2.3b (I)
23.0±3.2b (II)
48.0±4.1b (III)
20
36.0±3.5c (II)
59.0±6.3c (III)
69.0±5.5c (IV)
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30
53.0±5.2d (III)
71.0±7.6d (IV)
100.0±0.0d (V)
40
75.0±7.5e (IV)
100.0±0.0e (V)
100.0±0.0d (V)
50
100.0±0.0f (V)
100.0±0.0e (V)
100.0±0.0d (V)
*Repellency Class: class 0 (PR≤ 0.1%), class I (PR = 0.1–20.0%), class II (PR = 20.1–40.0%), class III (PR =
40.1–60.0%), class IV (PR= 60.1-80.0%), and class V (80.1-100.0%) [26], Cumulative mean±Sd followed by
the same letter within each vertical column are not significantly different (p<0.05).
Contact Toxicity
The lowest concentration of EO (6.0 µg/adult insect) had a mortality rate of 94.0±5.4% in 48 h while the
highest concentration (500.0 µg/adult insect) exhibited a mortality rate 100±0.0% in 12 h (p<0.05) (Table 4).
The activity of the EO was lower than that of pyrethrine (6.0 µg/adult insect) the positive control, which
attained 100% mortality within 12 h exposure period. The EO of the leaves of T. diversifolia exhibited contact
toxicity against S. zeamais adults insects, with LD50 values of 12.3 μg/adult insect at 95% upper and lower
fiducial limits (FL) (10.5 to 14.7 μg of oil/adult insect), for an exposure time of 12 h. The linear regression
equation of S. zeamais mortality after 12 h duration was y= 4.94x–0.355. This indicated that this EO was
approximately three times less toxic against S. zeamais than pyrethrine with a reported LD50 value of 4.29
μg/adult insect [21]. This EO was however, more toxic to S. zeamais than those obtained from Artemisia
giraldii with LD50 (40.51 μg/adult insect) and A. subdigitata with LD50 of 40.51 and 76.34 μg/adult insect
respectively, at an exposure time of 24 h [20].
Table 4. Mean cumulative percentage mortality for contact toxicities of essential oil from T. diversifolia
against S. zeamais (the maize weevil)
Conc (µg EO /Insect)
Percentage mortality
6 h
12 h
24 h
36 h
48 h
0(-ve control)
0.0±0.0a
0.0±0.0a
0.0±0.0a
0.0±0.0a
0.0±0.0a
6.0
12.0±5.5b
38.0±5.4b
44.0±5.5b
72.0±8.4b
94.0±5.4b
10.0
25.0±6.4c
46.0±6.2c
66.0±7.1c
96.0±4.0c
100.0±0.0c
17.5
42.0±8.4d
72.0±7.1d
100.0±0.0d
100.0±0.0c
100.0±0.0c
25.0
68.0±10.0e
100±0.0e
100.0±0.0d
100.0±0.0c
100.0±0.0c
6.0(+ve control)
86.0±6.2f
100.0±0.0e
100.0±0.0d
100.0±0.0c
100.0±0.0c
*Values on the table represent cumulative percentage mean±SD (N= 6). ** Means followed by different letters
in column are significantly different by Turkey’s test (p<0.05). –ve control = acetone, +ve control =
pyrethrine.
Monoterpenes including α-pinene, β-pinene and limonene, have been reported to exhibit fumigant activity,
antifeedant, insect repellents and growth regulatory activities against numerous pests, including S. zeamais
(maize weevil) and Callosobruchus maculatus (bean weevil) [27, 28]. Sesquiterpenoids such as α-copaene,
germacrene D, γ-cadinene and δ-cadinene, have also been reported to possess insecticidal properties.
According to Choi et al. [29], the toxicity of EO for stored-product insects could be influenced by their
chemical composition. Therefore, insecticidal activities of T. diversifolia EO could be associated with
components including α-pinene, β-pinene, and caryophylene among other constituents. These three
components accounting to approximately 70.6% of the oil could be responsible for these properties since they
have been reported to possess insecticidal activities against several insect pests including aphids, termites and
beetles [29, 30]. Resistance development in insects can be reduced by the use of essential oils due to
synergistic action between different molecules of the oil [31].
CONCLUSIONS
The essential oil of T. diversifolia as determined by GC/MS mainly comprises of monoterpenes and
sesquiterpenes, with α-pinene, constituting the greatest percentage. This oil exhibited strong fumigant,
repellency and contact toxicity activities against S. zeamais adult insects and thus has great potential of for use
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a potent insecticide. For the practical application of these essential oil as novel insecticide, further studies on
the safety of the essential oil to humans and isolation and identification of the active constituents in this
essential oil are necessary. In addition, studies of individual and combined activities of the different
components in the oils are necessary to ascertain whether the compounds act individually, synergistically or
antagonistically.
ACKNOWLEDGMENTS
The author appreciates the technical staff in the Departments of Botany and Zoology in Maseno University, for
authentication of plant materials and rearing insects for pesticidal activity assays of T. diversifolia.
DECLARATION OF INTEREST STATEMENT
The author reports there are no competing interests to declare.
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