International Journal of Trend in Scientific Research and Development (IJTSRD)

Volume 4 Issue 4, June 2020 Available Online: www.ijtsrd.com e-ISSN: 2456 – 6470

Insecticidal Activity of Stem Bark Extract of Lophira Alata

(Ekki) Against Cowpea Bruchid (Callosobruchus Maculatus)
Ifelolu A. Remi-Esan1, Olusola O. Bankole2
1Environmental
Biology Unit, 2Biochemistry Unit,
1,2Department of Science Laboratory Technology, Federal Polytechnic, Ilaro, Ogun State, Nigeria

ABSTRACT How to cite this paper: Ifelolu A. Remi-

Methanolic extract of Lophira alata was evaluated for its efficacy as contact Esan | Olusola O. Bankole "Insecticidal
and fumigant insecticides on cowpea bruchid, Callosobruchus maculatus in the Activity of Stem Bark Extract of Lophira
laboratory at ambient tropical conditions of temperature and relative Alata (Ekki) Against Cowpea Bruchid
humidity. The plant powder tested was applied at rates 0.0 (control), 2.0 g and (Callosobruchus Maculatus)" Published in
3.0 g / 20 g of cowpea seeds either directly for contact with the insect pest or International Journal
in plastic containers to assess its fumigant toxicity. Results of contact toxicity of Trend in Scientific
assay showed that powders of L. alata was effective against the adult C. Research and
maculatus causing 90 % mortality (4.00 ± 0.57) within 2 days of application at Development
3.0 g / 20 g of cowpea seeds as compared with 90 % mortality (5.38 ± 0.50) (ijtsrd), ISSN: 2456-
recorded on day 4 of 2.0 g concentration application. The results of fumigant 6470, Volume-4 |
assays showed that L. alata had the highest insecticidal activity causing 95 % Issue-4, June 2020, IJTSRD31207
mortality of C. maculatus within 4 days of application at rate 3.0 g / 20g of pp.776-781, URL:
cowpea seeds in contrast to 80 % mortality recorded in 96 hrs of 2.0g www.ijtsrd.com/papers/ijtsrd31207.pdf
concentration application. The phytochemical screening of the plant revealed
alkaloids, saponins, glycosides, phytosterols, tannis, flavonoids and terpenoids Copyright © 2020 by author(s) and
while reducing sugar was absent. This study showed that the tested plant International Journal of Trend in Scientific
product is toxic to cowpea bruchid and the powders can be mixed with Research and Development Journal. This
cowpea seeds to prevent hatching of the eggs thereby helping in their is an Open Access article distributed
management. under the terms of
the Creative
KEYWORDS: Callosobruchus maculatus, Lophira alata, Fumigant, Cowpea, Commons Attribution
Insecticidal License (CC BY 4.0)
(http://creativecommons.org/licenses/by
/4.0)
INTRODUCTION
Cowpea is one of the most significant pulse crops local to cowpea seeds are used for preparing delicasies for
Africa. Cowpea is called vegetable meat because of the high consumption purposes. In Nigeria, cowpea is associated to
measure of protein in grain with better organic incentive on pestivorus attack during pod and dry seed stages in the field
a dry weight premise. On a dry weight premise, cowpea during cultivation and in storage after harvesting. C.
grain contains 23.4 percent protein, 1.8 percent fat, and 60.3 maculatus has caused huge weight reduction and reduced
percent sugars and it is rich in calcium and iron. Aside from market value of cowpea seeds [4]. It has been accounted for
this, cowpea structures brilliant scrounge and it gives a that both quantitative and subjective loses emerging from
substantial vegetative development and spreads the ground physical, chemical and biological factors happen during
so well that it checkmates the soil erosion disintegration. As stocking of grains in different forms of facilities and stores
a leguminous crop, it fixes around 70 – 240 kg for each ha of [4].
nitrogen every year [1]. Cowpea is chiefly cultivated in
tropical and sub-tropical areas for its vegetable, seed and to The control of insect pests of stored grains and pulses are
lesser degree as a fodder crop. It is an adaptable crop in view achieved through use of synthetic chemical insecticides, such
of its covering nature, drought resistant nature, soil as methyl bromide. Several nations have however banned
reestablishing properties and multi-consumption methods. the usage due to environmental concerns and health hazards
As a pulse, cowpea fits well into a large portion of the associated to human consumption of these grains [5]. The
trimming frameworks of cropping [1]. The current day adverse effects of chemical insecticides have led researchers
cultivars display lower productivity, non-simultaneous to try to find new avenue of insect control, which has led to
blooming and fruiting, non-reaction to high portions of the discovery of plant products as an alternative way of
sources of organics like composts and fertilizers, water controlling insects [6,7]. Moreover, tropical regions are
system, culturing, resistance to major insect pest and believed to be endowed with many plant species with
diseases which cause considerable damage and very poor insecticidal properties and some of them are with medicinal
harvest [1]. The cowpea bruchid is planted and found properties [1]. Therefore, this research investigates the
everywhere, with its underlying invasion beginning in the insecticidal activity of oil extracts Ekki (Lophira alata)
field not long before harvest and are conveyed into the store against C. maculatus which is an important insect pests of
where it quickly increases [2,3]. Ripe and matured dry cowpea in storage.

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Materials and methods Phytochemical screening
Study area Phytochemical analysis and screening was carried out using
The study was conducted in the Zoology Departments simple chemical tests to detect the presence of secondary
Research Laboratory, University of Lagos and Pharmacology plant constituents such as alkaloids, tannins, flavonoids,
Department Laboratory, Lagos University Teaching Hospital saponins, triterpenes, sterols, phenols, glycoside, reducing
Lagos. sugar and soluble carbohydrate in the sample was carried
out as described by the general method of [12].
Insect culture
The insects used to establish a laboratory colony of C. Test of contact toxicity of plant extract on adult
maculatus came from a batch of infested cowpea seeds, mortality, oviposition and progeny development of
Vigna unguiculata (L.) Walp variety Ife brown collected from Callosobruchus maculatus
Nigeria Stored Products Research Institute Lagos, Nigeria. Plant extract of L. alata was admixed with cowpea seeds at
Beetles were reared subsequently by replacement of the rates of 2.0g / 10g of cowpea seeds in 250ml plastic
devoured and infested cowpea seeds with fresh un-infested containers. Ten pairs of adult C. maculatus (2 to 3 days old)
cowpea seeds in 2-L kilner jars covered with muslin cloth to sexed according to the methods described by [13] were
allow air circulation. Insect rearing and the experiments introduced into the treated. Untreated cowpea seeds were
were carried out at ambient temperature of 28 + 2 oC and 75 similarly infested. Four replicates of the treated and
+ 5 % relative humidity. untreated controls were laid out in Complete Randomized
Block Design in insect cage. Insect mortality was assessed
Plant collection every 24 hours for four days. Adults were assumed dead
The plant evaluated in this work was Lophira alata (Stem when probed with sharp objects and no response is made. At
bark). It was obtained in fresh form, free of insecticides from the end of day 4, all insects, both dead and alive were
LUFASI Nature Park, Eti-Osa, Lagos State, Nigeria and removed from each container.
authenticated by the Botany Department, University of
Lagos, Lagos State. The plant materials were rinsed in clean Test of plant extract as fumigant insecticide on adult
water to remove sand and other impurities, cut into smaller mortality, oviposition and progeny development of C.
pieces before air dried in a well-ventilated laboratory for maculatus
four weeks and ground into very fine powder using a Ten grams of the cowpea seeds were weighed into 50ml
mechanical blender. The powder was packed in plastic transparent plastic tubes that had been cut opened at the
containers with tight lids and stored in a refrigerator at 4 oC bottom and sealed with muslin cloth. Plant extract of L. alata
prior to use. weighing 2 and 3g concentrations were put into another
half-cut 25ml plastic tubes. The 50ml tube and 25ml tube
Collection of cowpea seeds were then joined together with the aid of gum [14]. Ten pairs
Cowpea seeds used for this study were obtained from a of adult C. maculatus (2 to 3 days old) sexed according to the
newly stocked seeds free of insecticides at Nigeria Stored methods described above were introduced to the tube
Products Research Institute Lagos State, Nigeria. Firstly, the containing 10g of cowpea seeds and tightly sealed [14].
seeds were cleaned and disinfested by keeping at -5 oC for 7 Untreated cowpea seeds were similarly infested. Four
days to kill all hidden infestations. This is because all the life replicates of the treated and untreated controls were laid out
stages, particularly the eggs are very sensitive to cold [8]. in Complete Randomized Block Design in insect cage.
The disinfested cowpea seeds were then placed inside a Bruchid mortality was assessed every 24 hours for four days.
Gallenkamp oven (model 250) at 40 oC for 4 hours [9] and Adults were assumed dead when probed with sharp objects
later air dried in the laboratory to prevent mouldiness [10] and made no responses. The experiment was kept inside the
before they were stored in plastic containers with tight lids. insect cage for another 30 days to allow for the emergence of
the first filial (F1) generation. The number of adults that
Extract preparation emerged from each replicate was counted with an aspirator
Hydroalchoholic extracts of L. alata was carried out using and recorded. The percentage adult emergence was then
cold extraction method. 730 g of the powder was weighed calculated using the method described by [13].
using an analytical balance (OHAUS, model number: PA124),
and soaked separately in an extraction bottle containing 5 % Progeny development = No of adult emerged x 100
liters of absolute methanol and water in the ratio 8:2. The No of eggs laid 1
mixture was stirred occasionally with a glass rod and
extraction was terminated after 72 hrs. The resulting Statistical Analysis
mixture was filtered using a double layer muslin blanket and Statistical Package for Social Science (SPSS, version 23) was
the solvent was evaporated using a rotary evaporator used in the analyses of the quantitative data. Data were
(EYELA, model number SB-1300) at 30 to 40 oC with rotary subjected to one-way analysis of variance (ANOVA). Where
speed of 3 to 6 rpm for 8 hrs. The resultant moisture solid significant differences existed, treatment means were
was transferred into a desiccator for 24hrs to remove any separated using the Tukey test and considered significant at
residual moisture, until a dry weight was achieved. The solid P ≤ 0.05.
extract was ground into a fine powder using a mortar and
pestle and the powder was stored at 40C in the refrigerator
prior to further use. [11].

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Results
Phytochemical screening
The phytochemical analysis of the methanolic extract of Lophira alata stem bark reveals that the plant has alkaloids, saponins,
glycosides, phytosterols, tannis, flavonoids and terpenoids while reducing sugar was absent as shown in Table 1 below.

Table 1: Result of phytochemical screening of the methanolic extract of Lophira alata stem bark
Phytoconstituents Lophira alata
Alkaloids +
Reducing sugar -
Saponins +
Glycosides +
Phytosterols +
Tannis +
Flavonoids +
Terpenoids +
Cyanide -
“+” represent class of phytochemicals present
“-” represent class of phytochemicals not present

Table 2: shows the percentage mortality, standard error of mean of adult Callosobrochus maculatus treated with
Lophira alata powder at rate 2.0 g / 20 g and 3.0 g / 20 g of cowpea seeds for contact toxicity.
Mortality % ± SEM after
Treatment
24hrs 48hrs 72hrs 96hrs
2g/20g 0.75 ± 0.25b 1.38 ± 0.18b 2.50 ± 0.19b 5.38 ± 0.50c
3g/20g 3.38 ± 0.46c 4.00 ± 0.57c 2.00 ± 0.68b 0.63 ± 0.38b
t-value 5.01 4.41 0.71 7.62
p-value 0.02* 0.07* 0.02* 0.34*
Control 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a 0.00 ± 0.00a

Each value is a mean ± standard error of four replicates. Means within the same column, with different
letters are significantly different at P < 0.05 using Tukey’s test.

Effectiveness of plant powder as contact insecticide at rate 2.0 g / 20 g of cowpea seeds

Figure 1 presents the effect of plant powder on mortality of C. maculatus. Within 24 - 48 hrs of application. The plant powder
was less potent on the insects, causing 25 % mortality and at 72 hrs, about 35 % insect mortality was also recorded. The plant
powder had a significantly high mortality effect on the insects at 96 hrs (Day 4), killing 90 % of the tested insects.

Figure 1: Percentage mortality of adult C. maculatus treated with powder of L. alata at 2.0 g / 20 g cowpea
seeds for contact toxicity

Effectiveness of plant powder as contact insecticide at rate 3.0 g / 20 g of cowpea seeds

Figure 2 presents the effect of plant powder on mortality of C. maculatus. The plant powder had a significantly high mortality
effect on the tested stored product insects at 24 - 48 hrs (Days 1-2), killing 90 % of the insects. However, the insect biomass
reduced as shown in the graph (Figure 2).

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Figure 2: Percentage mortality of adult C. maculatus treated with powder of L. alata at 3.0 g / 20 g cowpea seeds
for contact toxicity

Fumigant effect of plant powder of Lophira alata on adult Callosobrochus maculatus at rate of 2.0 g / 20 g and 3.0 g / 20
g cowpea seeds
At day 4 after post treatment, L. alata was able to cause 80 % of mortality of adult C. maculatus applied at 2.0 g / 20 g, days 1, 2,
and 3 had less mortality ranging from 15 %, 20 %, and 25 % respectively. The toxicities of the plant powder to cowpea bruchid
increased with an increase in concentration and period of exposure to the plant powder as shown in figure 3 below.

Bruchid mortality was rapid and highest at early expose rate of 3.0 g / 20 g on cowpea with about 95 % mortality recorded
between 48hrs exposure time. While the powder remains very active and potent due to its increased concentration as shown in
figure 4. The insect biomass had reduced before the day 4 (96 hrs) limit.

Figure 3: Percentage mortality of adult C. maculatus treated with powder of L. alata at 2.0 g / 20 g cowpea
seeds as fumigant insecticides

Figure 4: Percentage mortality of adult C. maculatus treated with powder of L. alata at 3.0g / 20g cowpea seeds as
fumigant insecticides.

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Discussion The high mortality and low progeny development caused by
Botanically derived insecticides are good alternatives to the powder of L. alata can be attributed to strong choky
popularly used synthetic chemical insecticides as many of odour disrupting respiratory activity of the beetles. The
them have often times yielded positive results when been results obtained from this study agree with those reported
used against insect pests of stored products, belonging to, by [29] in studies with six Citrus species peel oils against C.
but not limited to Coleoptera and Lepidoptera [15]. They are maculatus, S. zeamais and Dermestes maculatus.
known to be easily biodegradable and not toxic to none
target organisms which oftentimes are species of flora and In this study, the lethal effect of plant powder on the cowpea
fauna. The discovery of organochlorine and bruchid could be as a result of contact toxicity. Insects
organophosphate chemical insecticides in the late 1930s and breathe by means of trachea which usually opens at the
early 1940s, botanical insecticides have remained an surface of the body through spiracles [10]. These spiracles
important weapon in the farmers armory in managing insect might have been blocked by the powder thereby leading to
pests of their farm produce and consumables [16]. Many suffocation. The powder also prevented oviposition and
Nigerian plant species are medicinal and are proven to be progeny development when applied as contact insecticides.
effective against a wide range of insect pests [17-19]. The choky effect of these powder also disrupt mating
activities, sexual communication and inhibit locomotion an
In this current study, the methanolic extract of Lophira alata effect that have been reported by many researchers [30-32].
stem bark show a high effectiveness, it caused 100%
mortality of C. maculatus within 48 hrs and 96 hrs of Conclusion
application at all concentrations of 2.0g / 20g and 3.0g / 20g This research has proven that methanolic powder extract of
of cowpea seeds. The powder may have also blocked the L. alata is a good and cheap means to control insect pests of
spiracles of these insects which resulted in suffocation. This stored products, especially those insects in the order
results agree with the previous studies in which powders Coleoptera which have characteristic biting and chewing
and oils of A. indica, Z. zanthoxyloides, A. occidentale and M. mouthparts and are clumsy fliers. Therefore, can be
oleifera have been used as botanicals against different life integrated into other insect pest management systems for
stages of storage insects [20, 1]. effective pest control.

The preliminary phytochemical screening of methanolic Acknowledgement

extract of Lophira alata stem barks revealed the presence of The authors are grateful to Dr F. Adetoro of Applied
alkaloids, tannins, saponins, phytosterols, flavonoids, Entomology and Pest Management Unit, Zoology
triterpenoids and glycosides. Alkaloids, flavonoids, saponins, Department, University of Lagos for his timely intervention
triterpenes and tannins have been reported to possess in providing details on the identification and sexing of the
central nervous system modifying activity such as Bruchid used.
depression, sedation, anxiolysis, psychotropic, analgesic and
anti-convulsant activity [21-23]. The toxicity of L. alata to References
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