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INSECTICIDAL ACTIVITIES OF TWO MELIACEOUS PLANTS AGAINST Trogoderma

granarium Everts (Coleoptera: Dermestidae)

Article · January 2012

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I.J.S.N., VOL. 3(3) 2012: 696-701 ISSN 2229 – 6441

INSECTICIDAL ACTIVITIES OF TWO MELIACEOUS PLANTS AGAINST

Trogoderma granarium Everts (Coleoptera: Dermestidae)
Abdalla Abdelrahim Satti & Mokhtar Mohamed Elamin
Department of Alternatives to Pesticides and Biocontrol, Environment and Natural Resources Research Institute, National Centre for
Research, P.O. Box 6096, Khartoum, Sudan
E-mail: satisattisat@yahoo.com; Mobile: 00249 916017797

ABSTRACT
The khapra beetle (Trogoderma granarium) is one of the major economic store pests in Sudan. Application of chemical
insecticides, either in form of fumigation or spraying, is the sole measure of control adopted. Since these chemicals were
linked with several drawbacks, attempts are in progress to find environmentally sound and economically feasible
alternatives. Therefore, laboratory experiments were carried out to evaluate the insecticidal activities of several extracts
prepared from two meliaceous plants, viz., Khaya senegalensis (mahogany) and Azadirachta indica (neem), against the 3rd
instar larvae of T. granarium. The results showed variable insecticidal activities by the tested extracts. Mahogany leaves
exerted better actions than those of the neem leaves. Nevertheless, all leaves extracts manifested relatively low and delayed
mortalities as compared with neem seeds. Accordingly, the neem seeds hexane extract (oil) at 5%v/v, was the best
treatment induced significant knock down effect on the larvae 48hrs post treatments. It showed a progressive increase in
effectiveness with time to attain 92.5% mortality on the third week of exposure. This extract also repelled the pest (85.4%)
and saved sorghum seeds (45.5%) significantly as compared with the untreated control. Moreover, in spite of what has
been mentioned about the low mortality effect of mahogany leaves water extract, it recorded the superior saving of
sorghum grains (54.5%) suggesting the presence of potent repellent and/or antifeedant active constituents in this extract.
Therefore, additional studies are required to ascertain the actual bioactivities in mahogany leaves, and to proceed forwards
in formulating botanical insecticides from the neem oil.

KEYWORDS: Mahogany; neem; Trogoderma granarium; water extract; organic extracts.

INTRODUCTION numbers were now approaching ten million trees.

Pests control in most tropical countries, as in many other However, another meliaceous species of economic
regions of the world, depends largely on the usage of importance is the mahogany tree, Khaya senegalensis
conventional pesticides. But, due to several drawbacks (Des.) A. Juss., which is abundant along rivers and
linked with these chemicals, including mainly the seasonal streams especially in western and southern parts
development of resistant pest strains, toxicity to man and of the country (El Amin, 1990 and Bein et al., 1996). This
animals, environmental pollution and the increasing cost species is also known to contain active ingredients with
of pesticides, hence the search for new alternatives was some biological activities (Bamaiyi et al., 2006 and
thought to be important. Botanical pesticides were Ibrahim et al., 2006), but it has got little research as
considered among important promising tools attempted compared to the neem tree. Although, both of the
and showed compatibility with other tactics for storage foregoing trees are utilized for many socio-economic
pests’ management (Ahmedani et al., 2007). Although purposes, particularly as timbers, but no real exploitation
natural products were recognized since ancient times, their is recognized regarding their rich phytochemical
usages have been suspended after the appearance of constituents. Therefore, in the course of studies aiming to
synthetic pesticides. Examples of plants exhibiting find potent plants for preparing natural pesticides, the
pesticidal properties are numerous starting with current research was intended to evaluate and compare the
Chrysanthemum, Nicotiana and Derris species in earlier insecticidal activities of different extracts prepared from
times and ending with neem extracts in recent decades neem and mahogany trees. However, the 3rd instar larvae
(Whitehead and Bowers, 1983; Schmutterer, 1995). of the Khapra beetle (Trogoderma granarium Everts) were
In the Sudan, various studies were performed to evaluate used in the bioassays, as test insects.
the insecticidal activities of different indigenous plants for
controlling agricultural pests of field crops and store MATERIALS AND METHODS
products (Siddig, 1991; Satti et al., 2003; Sir El Khatim, Plant materials used
2005; Yousif and Satti, 2008). Among these plants, extract All plants parts used in this study were collected from
of neem (Azadirachta indica A. Juss.) tree (Meliaceae) has various locations in Khartoum State, during different times
received the greatest attention during the last decades as between 2009 and 2011. They included leaves of
potent source for biologically active ingredients. The neem mahogany tree (K. senegalensis) and leaves and seeds of
tree is widely distributed in the country, and its total the neem tree (A. indica). The neem fruits were already
stored in the laboratory since 2009. They were collected

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 Insecticidal activities of two meliaceous plants against Trogoderma granarium

from the ground after being dropped under neem trees many times to complete the required number of larvae in
during the fruiting period (June-August). However, the some experiments.
leaves of both tree species were picked fresh just one week Evaluation of mortality and repellent effects
before the commencement of the bioassay experiments, Three experiments were conducted successively to
and dried under shade. All samples were ground into fine compare the insecticidal actions (mortality effects) of the
powders using an electric blender (Moulinex) and kept in previously prepared treatments against the 3rd instar larvae
dark bottles until being extracted. of T. granarium, as test insects. The first experiment was
Preparation of botanical treatments and test insects conducted in January 2010 (winter season) to test the
Regarding organic solvent extractions, three solvents (viz., water and organic leaves extracts of mahogany and neem,
hexane, ethyl acetate and methanol) were used and then supported by another experiment during summer
successively in a soxhlet apparatus to extract the apolar period in May 2010 using merely leaves water extracts.
(oil), intermediate and polar compounds in neem seeds, The third experiment in February 2011 was dealt with
respectively. A weight of 20 grams seeds powder was neem seeds organic and water extracts. However, the
loaded on the soxhlet thimble and extracted to 9 hours to means room temperatures (maximum and minimum) and
free it from oil. The process was repeated 3 times with relative humidity were recorded for the study period. All
new samples to get the required quantity. Upon the above experiments were executed separately in Petri
completion of each round, the filtrate was air dried and re- dishes. According to the number of treatments, each 10
extracted successively by the ethyl acetate and the grams of clean sound sorghum (S. bicolor) grains were
methanol to obtain the other components of the sample, in treated with one of the different extracts concentrations,
the same way. However, the leaves samples of the two replicated four times, left for five minutes to dry, and then
plants were extracted only with ethyl acetate and methanol placed into the Petri dishes. Ten 3rd instar larvae of T.
in the same sequence. All extracts were kept separately in granarium were introduced in each Petri dish, including
dark bottles and placed in refrigerator until used. the untreated control, and covered. All experiments were
However, any plant sample was re-weighed at the end of assigned in Completely Randomized design. However,
each extraction step so as to determine the percentages of inspections of the Petri dishes were carried out
the extracted materials in relation to seed weight. periodically on the 2nd, 7th, 14th and 21st day following
Moreover, the extract obtained was also weighed for more treatments. Hence, the number of dead insects and other
confirmation. Water extraction was done for all the above observations were recorded. This allowed to evaluate the
mentioned plant materials of the neem and mahogany knock down and delayed effects of different treatments.
trees. Plant powders were prepared one day before the The collected records were tabulated and statistically
start of each experiment. Accordingly, the amount of analyzed. Whenever significant differences were achieved,
powder needed was weighed in a conical flask where half means separation was performed using Duncan's Multiple
volume of water was added, mixed thoroughly with a glass Range Test. The best botanical treatment (i.e., 5% neem
rod and allowed to stand overnight. In the next day the seed kernel hexane extract) that achieved from mortality
mixture was agitated manually for few seconds before experiments was selected for repellent test against the
filtration, using fine mesh. The volume of the extract was same pest. A locally made repellent apparatus was
completed with water to attain the highest concentration prepared and utilized in this study as described by Berndt
(5%w/v) indicated for the study. Consequently, the other (1963), but slightly modified in the control hole of
concentrations (2.5 and 1.25% w/v) were prepared through platform to accommodate a Petri dish 5cm in diameter.
serial dilutions. Preparation of treatments was performed The experiment was carried out in April 2011. Sorghum
as a pre-step for biological assays in each experiment. grains treated with 5% neem seed kernel hexane extract, in
Therefore, the intended organic extracts were firstly addition to the untreated control were placed randomly in
diluted with water to prepare the highest extract the peripheral holes of the repellent apparatus. A number
concentration (5%). However, in case of oil extract, 0.1% of 200 3rd instar larvae of T. granarium were introduced in
liquid soap was added as an emulsifier. Consequently, the a Petri dish located in the central hole of the platform, then
other concentrations (2.5 and 1.25%) were prepared by closed by a glass cover provided with a glass rod fitted
serial dilutions with water. On the other hand, the three into the top opening cover of the equipment. After a
rates of water extracted materials were prepared on the moment, the glass rod was pulled up to permit releasing of
same day following the extract preparation as shown the insects. In the second day (24 hours) the number of
previously. The different extracts concentrations were thus insects detected in each peripheral hole was recorded.
ready to be evaluated in bioassay experiments. Based on Such experiment was repeated successively in subsequent
these experiments, the concentration which reflected the two days, following the same procedures. Hence, three
best mortality performance was selected for a repellent counts were taken and analyzed to calculate the
test. The number of larvae allocated for each experiment attractancy or repellency effect of the treatment according
was segregated from the culture with the help of a small to Leonard and Ehrman (1976) formula, as shown below:
camel hairs brush, white A4 paper sheet and a Petri dish.
This was done each time by taking some amount of A=
infested grains from the jar and spread carefully on the
sheet paper, then returned back again into the jar to expose
different stages of larvae clinging to the paper. Using the Where; A = attractancy (+) or repellency (-); No = number
brush, only the 3rd instar larvae were dropped from the of insects in the test hole; Nb = number of insects in the
sheet down into the dish. The process may be repeated control hole; Nt = the total number of insects in both
holes. The output of this equation ranges from +1 (100%

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I.J.S.N., VOL. 3(3) 2012: 696-701 ISSN 2229 – 6441

attractant) to -1(100% repellent) when compared to the Regarding mahogany leaves, methanol (20.5%) and water
control. (17.4%) extracts gave the best significant yields. However,
neem seeds produced higher yields than the leaves extracts
Weight loss of sorghum grains of the two plants (mahogany and neem). Neem seeds
Weight loss of sorghum seeds according to larval damage hexane extract gave the highest significant yield (46.7%),
during the insecticidal bioassay experiment was calculated followed by neem seeds methanol extract (28.9%) and
for four selected treatments. These included; mahogany neem leaves methanol extract (16.9%). It is clear that in
leaves water extracts (5% w/v), neem leaves methanol both plants ethyl acetate extracts gave the lowest yields.
extract (5% w/v), neem seeds hexane extract (5% w/v) and These results suggested that the content of polar materials
the control (untreated sorghum seeds). Evaluation of in most tested plant parts were higher than the
damaged seeds was carried out 45 days following the intermediate polarity compounds. Also, the higher oil
treatments. Firstly, the insect larvae were removed from content in neem seeds was considered as an indicator for
Petri-dishes, then the infested grains were subjected to rich apolar constituents with diversified biological
sieving to get rid of cast skin, grain dust, insect excretion activities in this extract. However, literature showed
and other debris occurred due to insect feeding and variable levels of oil content in seeds which reported to
development. Lastly, Petri-dishes contents (sorghum contain numerous active ingredients (e.g. azadirachtin,
seeds) were weighed again for calculating the percentages salannin, nimbinen, salannol, salannolacetate, and stearic,
of damaged seeds based on the initial weights (10g/dish). oleic and linoleic acids) with pesticidal properties
(Schwinger, 1984 and Bashir, 1994). The oil level
RESULTS AND DISCUSSION achieved in this study (46.7%) agreed with Maydell (1986)
Extraction yields of plant samples who mentioned that the neem seed kernels contain about
The result of extraction yields (quantity and percent) of the 45 – 60% oil.
studied plant samples were explained in Table (1).

TABLE 1. Yields of extracted materials from different parts of mahogany (Khaya senegalensis) and neem (Azadirachta
indica) trees, using water and organic solvents.

Plant samples Extracted materials per 20 g

Mean±S.E. (%)
Ι. Mahogany:
M-L. eth.ac. extract 1.7±0.0b 08.7
M-L. meth. extract 4.1±0.1a 20.5
M-L. water extract 3.5±0.1a 17.4
C.V. % 5.6
IV. Neem:
N-S. hex. extract 9.3±0.0a 46.7
N-S. eth.ac. extract 2.0±0.0d 09.9
N-S. meth. extract 5.8±0.0b 28.9
N-L. eth.ac. extract 1.4±0.3e 07.0
N-L. meth. extract 3.4±0.2c 16.9
C.V. % 6.1

M-L.= mahogany leaves; eth.ac.= ethyl acetate; ext.= extract; meth.= methanol;
N-S.= neem seeds; hex.= hexane; N-L.= neem leaves.

Insecticidal effects of botanical treatments 31.2±4.6%R.H.). The highest concentration (5%) of both
Table (2) showed the mortality effects of mahogany and plants gave the best significant results since the second
neem leaves extracts on the 3rd instar larvae of day of treatment, with mahogany extract being superior at
Trogoderma granarium, as a result of winter experiment all intervals. Notably, summer experiment manifested
(average 28.6±1.4oC and 29.6±4.1%R.H.). No significant higher mortality rates than the winter experiment. Such
differences were detected between all plants extracts up to seasonal difference in results could be connected to
seven days post treatments. However, mahogany water variation in temperature degrees between the two seasons;
extract at 5% concentration reflected the highest a factor seems to affect the activity of the pest. It was
significant mortality means at 14 days interval (15.0% observed that, during hot times the larvae of T. granarium
mortality) and onwards, followed by 5% neem methanolic were activity feeding and crawling, while in cold they tend
extract. The rest of treatments exerted no significant to be more quiescent. Whatsoever, the superiority of
differences from the control. Similarly, Table (3) showed mahogany leaves water extract could be attributed either
the mortality results of leaves water extracts of the two to high concentration or more potent active ingredients in
plants which performed in summer time (36.7±0.2oC and this extract compared with the other plant.

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 Insecticidal activities of two meliaceous plants against Trogoderma granarium

TABLE 2. Mortality levels of Trogoderma granarium 3rd instar larvae as affected by mahogany (Khaya senegalensis)
and neem (Azadirachta indica) leaves organic and water extracts, during winter season (January 2010).

Treatments Mortality percent means (±S.E.) at different intervals

2 days 7 days 14 days 21 days
M-L. eth.ac.ext.1.25% 0.0±0.0a 00.0±0.0a 00.0±0.0c 02.5±0.2bc
M-L. eth.ac.ext. 2.5% 0.0±0.0a 02.5±0.2a 02.5±0.2c 05.0±0.3abc
M-L. eth.ac.ext. 5% 0.0±0.0a 00.0±0.0a 00.0±0.0c 07.5±0.5abc
M-L. meth.ext. 1.25% 0.0±0.0a 00.0±0.0a 00.0±0.0c 02.5±0.2bc
M-L. meth.ext. 2.5% 0.0±0.0a 00.0±0.0a 00.0±0.0c 00.0±0.0c
M-L. meth.ext. 5% 0.0±0.0a 02.5±0.3a 02.5±0.2c 05.0±0.3abc
M-L. water ext. 1.25% 0.0±0.0a 02.5±0.3a 02.5±0.2c 05.0±0.3abc
M-L. water ext. 2.5% 0.0±0.0a 05.0±0.3a 07.5±0.2abc 07.5±0.2abc
M-L. water ext. 5% 2.5±0.2a 10.0±0.4a 15.0±0.5a 15.0±0.5a
N-L. eth.ac.ext. 1.25% 0.0±0.0a 02.5±0.3a 05.0±0.5bc 05.0±0.5abc
N-L. eth.ac.ext. 2.5% 0.0±0.0a 07.5±0.3a 07.5±0.2abc 07.5±0.2abc
N-L. eth.ac.ext. 5% 0.0±0.0a 05.0±0.3a 07.5±0.2abc 10.0±0.4abc
N-L. meth.ext. 1.25% 0.0±0.0a 05.0±0.3a 05.0±0.3bc 05.0±0.3abc
N-L. meth.ext. 2.5% 2.5±0.2a 05.0±0.3a 07.5±0.2abc 07.5±0.5abc
N-L. meth.ext. 5% 0.0±0.0a 07.5±0.2a 12.5±0.2ab 12.5±0.2ab
N-L. water ext. 1.25% 0.0±0.0a 05.0±0.3a 05.0±0.3bc 05.0±0.3abc
N-L. water ext. 2.5% 0.0±0.0a 00.0±0.0a 00.0±0.0c 00.0±0.0c
N-L. water ext. 5% 0.0±0.0a 02.5±0.2a 05.0±0.3bc 05.0±0.3abc
Control 0.0±0.0a 00.0±0.0a 00.0±0.0c 00.0±0.0c
C.V. % 577.4 138.9 121.7 115.0
M-L.= Mahogany leaves; eth.ac.= ethyl acetate; ext.= extract; meth.= methanol; N-L.= neem leaves.

TABLE 3. Mortality levels of Trogoderma granarium 3rd instar larvae as affected by mahogany (Khaya senegalensis) and
neem (Azadirachta indica) leaves water extracts, during summer period (May 2010).
Treatments Mortality percent means (±S.E.) at different intervals

2 days 7 days 14 days 21 days

M-L. water ext. 1.25% 0.0±0.0b 00.0±0.0c 10.0±0.4b 10.0±0.4bc
M-L. water ext. 2.5% 0.0±0.0b 00.0±0.0c 10.0±0.4b 17.5±0.6ab
M-L. water ext. 5% 7.5±0.2a 17.5±0.2a 22.5±0.2a 27.5±0.5a
N-L. water ext. 1.25% 0.0±0.0b 00.0±0.0c 00.0±0.0c 00.0±0.0c
N-L. water ext. 2.5% 0.0±0.0b 00.0±0.0c 02.5±0.2bc 02.5±0.2bc
N-L. water ext. 5% 5.0±0.3a 05.0±0.3b 05.0±0.3bc 12.5±0. 5b
Control 0.0±0.0b 00.0±0.0c 00.0±0.0c 00.0±0.0c
C.V. % 157.3 92.5 78.4 69.3

M-L.= Mahogany leaves; ext.= extract; N-L.= neem leaves.

TABLE 4. Mortality levels of Trogoderma granarium 3rd instar larvae as affected by neem seeds organic and water
extracts, during February 2011.
Treatments Mortality percent means (±S.E.) at different intervals
2 days 7 days 14 days 21 days
N-S. hex.ext. 1.25% 00.0±0.0b 00.0±0.0b 02.5±0.2d 05.0±0.3f
N-S. hex.ext. 2.5% 02.5±0.2b 05.0±0.3b 12.5±0.5cd 22.5±0.5e
N-S. hex.ext. 5% 45.0±0.9a 80.0±1.2a 87.5±1.0a 92.5±0.5a
N-S. eth.ac.ext. 1.25% 00.0±0.0b 00.0±0.0b 12.5±0.2cd 27.5±0.5e
N-S. eth.ac.ext. 2.5% 00.0±0.0b 00.0±0.0b 10.0±0.4cd 32.5±0.5cde
N-S. eth.ac.ext. 5% 00.0±0.0b 07.5±0.5b 35.0±0.5b 47.5±0.5b
N-S. meth.ext. 1.25% 00.0±0.0b 00.0±0.0b 07.5±0.2d 20.0±0.4e
N-S. meth.ext. 2.5% 00.0±0.0b 02.5±0.2b 12.5±0.5cd 25.0±0.3e
N-S. meth.ext. 5% 00.0±0.0b 07.5±0.2b 30.0±0.4b 42.5±0.2bcd
N-S. water ext. 1.25% 00.0±0.0b 00.0±0.0b 10.0±0.4cd 30.0±0.4de
N-S. water ext. 2.5% 00.0±0.00b 00.0±0.0b 22.5±0.2bc 45.0±0.3bc
N-S. water ext. 5% 00.0±0.0b 05.0±0.2b 30.0±0.4b 52.5±0.8b
Control 00.0±0.0b 00.0±0.0b 00.0±0.0e 00.0±0.0f
C.V.% 123.7 77.4 40.6 24.2
hex.= hexane; ext.= extract; N-S.= neem seeds; eth.ac.= ethyl acetate; meth.= methanol.

According to Ibrahim et al. (2006), mahogany leaves reported that neem leaves contain moderate concentration
contain several active compounds such as saponin, of tannins.The effects of organic and water extracts of
phenols and tannins. On the other hand, Biu et al. (2009) neem seeds were explained in Table (4). Contrary to what

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I.J.S.N., VOL. 3(3) 2012: 696-701 ISSN 2229 – 6441

have been shown by leaves extracts, relatively higher the potent treatment against the pest. It reflected
knock down effects were exerted by seeds, particularly the significant effect with 85.4% repellency on the larvae.
hexane extract, 48hrs of treatments. As a result, the 5% However, the current results demonstrated that the latter
neem seeds hexane extract showed significant mortalities extract constitutes very good repellent as well as toxicant
at all investigation intervals, as compared with the other active ingredients against the tested insect. This result
treatments and the untreated control. It is obvious that this agreed with Nasr (1993) and Bakheet (2008) who assured
treatment showed gradual increase in mortality with time that neem oil has a very high repellency effects to T.
from the second day (45.0% mortality) up to the third granarium. According to Balandrin et al. (1988) the
week (92.5%). In the same context, Table (5) proved the repellent action of neem oil could result from the presence
repellency effect of neem seeds hexane extract (5%), as of volatile sulphur containing compounds.
.
TABLE 5. Repellency percent of neem seed hexane extract (5%) against the 3 rd instar larvae of Trogoderma granarium,
during April 2011.
Treatments Mean (±S.E.) insects/ Repellency percent
treatment. (%)
N-S. hexane extract 5% 5.0±0.6b 85.4
Control 63.3±5.7a
C.V. % 29.1

N-S.= Neem seeds.

Moreover, the results of sorghum seeds lost after 45days including mainly several triterpenoid compounds as
as a result of larval feeding on certain treatments were mentioned before (Butterworth and Morgan, 1968; Gill
shown in Table (6). The mahogany leaves water extract and Lewis, 1971 and Schwinger, 1984). Such high oil
(5%) followed by neem seeds hexane extract (5%), were constituent in neem seeds was considered as an added
the best treatments reduced sorghum grains damage value to its highest biological activities against the pest.
significantly compared with the untreated control. This This is particularly important whenever this plant
may be resulted from the multi-activities (viz, insecticidal, component is needed for commercial production of natural
repellency and antifeedant effects) induced by these insecticides.
extracts (Ahmed, 1993; Jood et al., 1993; Sharma, 1999
and Mohamed, 2003). However, the antifeedant actions of CONCLUSION AND RECOMMENDATION
mahogany and neem extracts were reputed to the presence The study revealed that extracts of mahogany, Khaya
of certain active compounds in the two plants (Schwinger, senegalensis, leaves were more effective than those of the
1984 and Ibrahim et al., 2006). However, the superior neem leaves in controlling the larvae of Trogoderma
saving of sorghum grains showed by the mahogany leaves granarium. Nevertheless, neem seed oil (5%v/v) was
water extract (54.5%) need to be investigated in further proved to be the most potent extract of the studied
studies. Although, inferior mortality result was shown by materials which reflected the best significant mortality
this extract, it may constitute powerful repellent and/or action against the pest. This extract also significantly
antifeedant effects, compared with those of the neem repelled the pest and saved sorghum seeds as compared
extracts. As shown previously, the different plant samples with the untreated control. But, due to superiority of
have yielded variable levels of extracted materials which mahogany leaves water extract in saving sorghum grains,
performed differently in the bioassay tests. The highest this extract required additional studies to clarify such
mortality and repellent effects exerted by neem hexane finding. Mean while, the neem seeds hexane extract (5%)
extract could be attributed to the variable active was advocated to be formulated as natural biocide.
ingredients reported in the ample oil content of the seeds,

TABLE 6. Sorghum grains (seeds) damaged by the 3rd instar larvae of Trogoderma granarium, after 45 daysfollowing
treatments with different plants extracts.
Treatments Seeds weight loss Seeds weight saved
Mean (±S.E.) (%) (%)
N-S. hexane extract 5% 0.6±0.0b 06.0 45.5
N-L. methanol extract 5% 1.0±0.0a 10.0 09.1
M-L. water extract 5% 0.5±0.0c 04.8 54.5
Control 1.1±0.1a 11.0 -
C.V. % 10.5
N-S.= Neem seeds; N-L.= neem leaves; M-L.= mahogany leaves.
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Sudan. safety. Pak. J. Agric. Sci. 44(3), 481 – 493.

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