CHAPTER ONE

1.0 INTRODUCTION
Protection of agricultural stored products against insect pests has proven to be of utmost
importance all over the world so as to achieve continuous and safe food supply. Plant extracts
contain biologically active compounds and hence has been the matter of interest for nearly
sixty years ago (Jiraungkoorskul, 2016). The phytochemicals are the major subject of interest
due to the fact that their large-scale synthesis and production for commercial use is immature.
This commercialization can only be achieved when immense knowledge of the
phytochemical components and their effect on the stored product as well as human health is
acquired (Caswell, 2019).
The demand for food increases with an ever-growing population. Thus, it is necessary to
protect crops and stored grains from pests. The multivoltine pest, cowpea weevil
Callosobruchus maculatus (Coleoptera: Bruchidae), causes significant damage to stored
pulses. It has been reported that C. maculatus alone can cause as much as 90% damage during
the three to six months of storage. Due to their potencies to cause lethality in most life stages
of a range of pests, synthetic pesticides are frequently used to protect both crop plants and
stored grains. Synthetic pesticides can adversely affect non-target organisms, including
humans, accumulate in the environment, pollute soil and ground water. Some of the synthetic
pesticides are also carcinogenic. The overuse of synthetic pesticides for insect control poses
risks to wildlife and even humans. Toxic potency of synthetic pesticides and their potential
effects have stirred interest from the public and regulatory agencies in alternative options for
pest management. Until the relatively recent development of synthetic pesticides and their
widespread application since the 1940s, phytochemicals have long been successfully used to
manage pests in crops. Extracts of plants and other secondary metabolites of plants,
microorganisms and enzymes are becoming increasingly popular as alternatives to synthetic
pesticides.
Bio-insecticides can be very effective, selective and have little potential for developing
resistance to target pests, as well as having minimal effects on non-target organisms. Cowpea
weevils is found as the most abundant bruchid that attacks and damage unpreserved cowpea.
Cowpea weevils (C. maculatus) is a cosmopolitan post-harvest pest of cowpea in the tropics.
It infest cowpea on field as well as on storage, rendering it unsuitable for consumption and as
they reproduce, great losses can be recorded (Jiraungkoorskul, 2016). The control of these
pest is achieved by mostly the used of pesticides for pest control despite the fact that small
scale farmers find it difficult to acquire due to financial or technical reasons. These pesticides
1
has also recorded negative effect on non-target animals especially on the field, other crops,
environment, and humans (Al-Moajel and Al-Fuhaid, 2016). Therefore, the development of
alternating, easy, safe and cheap control methods are needed for protecting stored cowpeas
against Callosobruchus maculatus. In developing an effective less toxic and environmentally
friendly method, attention is directed toward the use of natural products usually
phytochemicals of plants and their products in the control of food pests (Keita et al., 2021).
Most bioinsecticide researches were carried out on three biological responses; ovipositional,
feeding deterrent as well as contact and fumigant toxicity effect. Actual conditions of cowpea
storage are not taking into condition by most authors in laboratory bioassay. Long-term
storage condition that are close to possibly to on farm storage should be considered in a
controlled laboratory environment (FAO, 2015). In this review, we present a range of several
phytochemicals (mostly essential oils and alkaloids) of important plants and their toxic
effects on several control measures of cowpea weevils. We also present some active
compounds and their effect on stored grain as well as suitability for human health with
respect to the toxicity effect of the concentration of each extract used by various authors (Al-
Moajel and Al-Fuhaid, 2016; Roberts and Moreau, 2016; Al-Moajel and Al-Fuhaid, 2016;
Berger, 2018; Sharma et al., 2016).
1.1 Background of the study
Cassia occidentalis L. is an annual or perennial plant which is used in several traditional
medicines to cure various diseases. C. occidentalis is a spiny herb grows all over in Nigeria in
shade as well as under open condition. Generally found up to an altitude of 1,000 m in Africa
and wild throughout the plains on waste lands or in the coastal areas. It is also found in
deltaic region of western, eastern and southern African. Found particularly in the seacoast
throughout the hotter parts of Nigeria. It is also grown as an ornamental plant. This plant is
called in different regional/vernacular languages like Hausa (Sanga – Sanga), in Yoruba
( Ewe Asunwun), and in English Coffe sena, Rubbish cassia, stinking weed, Foetid cassia)
(Bhattacharyya et al., 2003).
In this review, an attempt was made to collect all possible ethnobotanical and nutraceutical
potential of C. occidentalis with reference to its food and medicinal applications. A baseline
survey was conducted during 2011-2015 and information’s about C. occidentalis were
collected through semi-structured interviews and discussion with the local healers, elderly
and experienced people. Additionally, all available literature on C. occidentalis was reviewed
and studied through online search engine Scopus and Google Scholar. Literature collection

2
was done from 1965 to 2015 and all the information were compiled and presented in this
review work (Dalziel, 2016; Al-Moajel and Al-Fuhaid, 2016).
C. occidentalisis a perennial plant which is erect and annual. The plant grows up to 2m tall.
The annual herb is soft wooded, foetid. The stems are greenish brown in colour and are
smooth in texture. The primary roots of C. occidentalisare diarch in nature and they have a
dicotyledonous secondary growth. Cross-section of the root shows the presence of a few
number of radical vascular bundle with exarch xylem. The leaves are pinnately compound
and alternate in nature. Each leaflet is 4 to 6cm long and 1.5 to 2.5cm wide and is green in
colour. The shape of the leaflets is ovate or oblong, lanceolate and has a pointed tip. The
leaflets have an entire leaf margin; the leaf apex is acuminate and has fine hairs on the
margins of the leaflets. There is the presence of a swollen cavity in the leaf base. On crushing
of the leaf, there is a fetid smell which is a characteristic of the herb (Hudaib et al., 2008).
Insect pests are a major constraint on crop production, especially in developing countries.
The cowpea weevil, Callosobruchus maculatus, F., (Coleoptera: Bruchidae) is a serious pest
of stored grains in sub-sahara Africa (Al-Moajel and Al-Fuhaid, 2016). Postharvest losses of
cowpea 3-4 months in storage caused by C. maculatus infestation have been reported as high
as 50% in Northern Nigeria (Caswell, 2019) and 60% in Northern Ghana (Tanzubil, 2019).
The loss of cowpea is a serious problem in Africa where as much as 20-50% of the grain is
damaged by C. maculatus (FAO, 2015). The damage of this magnitude is incredibly high and
demonstrates the destructive nature of the pest which can threaten food security at both
household and national levels. This is a major agricultural problem for farmers in developing
countries (Berger, 2018; Sharma et al., 2016).
Weevil infestation causes weight loss, quality deterioration resulting in overall
unacceptability in markets and impaired germinability of grains (Keita et al., 2021). Infested
grains are rendered unfit for consumption and sale. Consequently, farmers are compelled to
sell their products early after harvest when prices are still low partly because of anticipated
losses of the grain in storage. Over the years, the destructive activities and menace of storage
pests have been effectively suppressed with synthetic organochlorine and organophosphate
compounds like carbon disulphide, phosphine, malathion, carbaryl, pirimiphos methyl and
permethrin (Adedire et al., 2021). The application of these chemicals as pest control agents
is, however, fraught with problems, such as high persistence of the compounds, resurgence
and genetic resistance of pests, negative effects on non-target organisms, poor knowledge of
application, direct toxicity to the users, non-availability of the chemicals and increasing costs
of application (Berger, 2018; Sharma et al., 2016).
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1.2 Statement of the problem
Several studies on the efficacy of botanical pesticides for controlling insect pests are reported
extensively in cereal crops such as maize, beans, rice, cowpeas both in the field and during
storage. However, few studies on the use of botanical pesticides for controlling insect pests
on cabbage leafy green vegetable are reported. Specifically, little information exists on
insecticidal actions and synergistic effects of various botanical pesticides including their
derivatives on insect pests infesting cabbage leafy vegetable crops found specifically in
Nigeria. As a result, these traditional practices have prompted researchers to conduct
scientific study to verify many therapeutic effects that Cowpea is said to have. For this
reason, several extracts of the plant have been studied in order to test its antimicrobial,
antifungal activities and even its hepato-protective effect. At the same time, other researchers
have been interested in the qualitative chemical composition of plant extracts. This study was
undertaken to contribute to the evaluation of its phytochemical constituents and also, to
explore the antiradical and antimicrobial potentials of its various extracts.
1.3 Aim and Objectives
Aim
The aim of this study is evaluate of insecticidal effect of methanolic extract of spinach on
cowpea weevels.
Objectives
The objectives of this study are:
i. To determine the effect of methanolic extract of spinach on cowpea weevels.
ii. To evaluate of insecticidal effect of methanolic extract of spinach on cowpea weevels.
1.4 Justification of the study
It is common practice in traditional African communities to use locally available plants for
medicinal purposes and in agriculture (Obeng-Ofori et al., 2016). Natural plants products
possess insecticidal properties against a wide range of insect pests. For instance cowpea seeds
treated with cashew nut liquid (Echendu, 2019), fruit powder from Piper guineense (Ivbijaro
and Agbaje, 2016), certain spices (Igbai and Poswal, 2015), essential oil from sandbox (Hura
crepitans L.) (Ajayi and Adedire, 2016) and leaves of Eucalyptus (Berger, 2019) were better
protected than untreated seeds. Therefore, more investigations are necessary to explore the
natural protectants available within the locality for a more sustainable approach in controlling
storage pests.

4
1.5 Scope and limitation of the study
The limitation of this study was that of time limitation due to the study period and the
available resource. The importance of this research is also to add to existing knowledge.

5
 CHAPTER TWO
2.0 LITERATURE REVIEW
2.1 Cassia occidentalis
Cassia occidentalis, an annual herb, commonly known as chakwad or Wild Senna., belongs to
the 13 family Leguminosae. It is found all over India especially in the states of Rajasthan,
Uttar 14 Pradesh, Madhya Pradesh, Orissa, Jharkhand, Bihar. In Rajasthan it grows in semi
arid areas up 15 to a height of about 30 to 90 cm and has immense nutritional and medicinal
value. It occurs 16 along roadsides and in wastelands as a rainy season weed. Population
authentication has been 17 carried out of its samples from six states namely Uttarakhand,
Uttar Pradesh, Bihar, Jharkhand 18 and Odisha (Kumar and Roy, 2018). The plant Cassia
occidentalis can be regarded as a poor man’s food. 19 It contains proteins, a small amount of
fat, many minerals and natural fibers. Nutritional and 20 biological evaluation of Cassia
occidentalis seeds has been carried out by Mahajan (2013). Raw seeds of 21 Cassia
occidentalis contained 15 % protein, 1 % fat and 4 % fiber, 71 % carbohydrates. Some value
22 added food products namely coffee, chutney, breads of different types were prepared by
23 supplementing Cassia occidentalis seeds. These products were found acceptable as per the
organoleptic 24 evaluation (Mahajan, 2013). Leaves of Chakvad are traditionally used in
preparation 25 of “Sag” a leafy vegetable (Kar and Borthakur, 2008). The amino acid profiles
of the samples 26 revealed high protein quality of the vegetable (Kubmarawa et al, 2011).
29 Different parts of the plant (leaves, seed, and root) are known for their medicinal
importance 30 (Pawar and D’mello, 2011; Shadab et al, 2019). As such it is not only useful in
solving food 31 problems of the poor, it is also useful as a digestion and metabolism
corrective substance and as 32 a liver and cardio tonic. The dried seeds powder is given to
cure asthma (Punjani and Kumar, 2002). Cassia occidentalis seeds when fed to rats resulted
in a decrease in blood cholesterol . Seeds and 34 leaves were found to contain a number of
chemical constituents (Rai et al, 2000, Suradkar et al., 2017). Antinutritional factors like
phytic acid, trypsin inhibitors and tannins were also present in 36 seeds (Mahajan, 2013). No
work on nutritional analysis of leaves samples from Rajasthan has 37 been reported till date.
Hence leaves were analysed from different areas of Rajasthan so as to 38 study the variation
in nutrients and identify the region with maximum nutrient content.
2.1.2 Scientific classification Cassia occidentalis:
Botanical name - Cassia occidentalis
Family name - Caesalpiaceae
Kingdom - planatae
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Division - Magnoliophyta
Class - Magnoliopsida
Order - Fabales
Family - Fabaceae
Sub-tribe - Cassiinae
Genus - Cassia
Species - tora
2.2 BOTANICAL DESCRIPTION
Leaves are evergreen, lanceolate, compound, glossy leaf surface, deep tap root system, stem
is hard and woody, Dicot seed type with characteristic odour and bitter taste. Flowers are
yellow in colour with 1 to 2 cm diameter arranged in raceme type inflorescence, axillary and
also forming terminal, bracts are caduceus. Fruits are flat pods, 10-12 cm long with 10-30
seeds. Areolate seeds are pointed at end and blunt at the other (Evans and Trease, 2000).
2.3 PLANT DESCRIPTION
Cassia occidentalis Linn, usually grows in the southern part of Nigeria which is known as
Kasmard in Sanskrit, Kasondi in Hindi and C. occidentalisin English. The plant belongs to
Caesalpiniaceae family. The common name is Ponnavarai in Tamil. The roots, leaves and
seeds are the parts of the plant used. It is an erect herb, commonly found by road sides,
ditches and waste dumping sites. Cassia occidentalis has been widely used as traditional
medicine. Entire parts of the plant have medicinal values (GajaLakshmi et al., 2012).
Leaves:
The transverse section through the midrib region shows a plano-convex outline. There is a
single vascular bundle in the centre in which the phloem is more developed on dorsal side;
xylem is represented mostly by vessels, tracheids and thin walled parenchyma. It is also
exhibits a dorsal ventral structure. The upper epidermis is a single layered, covered externally
with moderately thick cuticle and 1-4 cellesed non-glandular trachoma’s. Rubiaceous
stomata are more abundant on upper surface than the lower ones.
Fruits and seeds:
The transverse section of seed shows the seed coat consisting of longitudinally elongated
cells followed by palisade layer compose columnar cells and parenchyamatus forming inner
layer of the testa in which a few vascular bundles are scattered. Embryo consists of radical,
plumule and two cotyledons, epidermis of cotyledons consist of single externally covered
with cuticle followed by two layers of palisade like cells of mesophyll. A mesophyll of

7
ventral side composed of polygonal cell filled with round to oval starch grain, vascular
bundles and few rosette crystal of calcium oxalate.
Root:
The transverse section of shows the single layered epidermis composed of cubical cells.
Epidermis is followed by 3-4 layers of parenchymatous cortex shows distinct endodermis
with casparian dots on the radial walls.
Habitat:
The plant of C. occidentalisis native to Bahamas, Barbados, Belize, Cayman Islands, Mexico
and other regions of North American continent. Also in the South American subcontinent, it
is indigenous to Argentina, Brazil, Ecuador, Colombia, Paraguay, Peru and other regions. C.
occidentalisis introduced in various other countries like Egypt, Angola, Madagascar,
Mauritis, Mali, Hawaii, Papua New Guinea and various other countries. This plant is spread
widely throughout the continent of Asia. It can be found in Bangladesh, Bhutan, China,
Brunei, Hong Kong, Nigeria, Iran, Iraq, Japan, Nepal, Pakistan and other regions. In the
Nigerian subcontinent Cassia occidentalis grows up to an altitude of 1500 m. It can be found
in Andhra Pradesh, Sikkim, Lakshadweep, Andaman and Nicobar Islands and in Gujarat
(Tagboto and Townson, 2001).
2.4 Ecological Requirement of Cassia occidentalis:
Cassia occidentalis and other s Cassia species are annual under shrub grows all over the
tropical sub-tropical countries and grows well in a wasteland as rainy season weeds. It is also
Cassia occidentalis grows in low lying coastal area, river banks, abundant in a waste places
and others moist places like uncultivated fields.
The Cassia occidentalis Species have been identified as the weeds that were withstand of
various environmental changes or factors and also well grows and abundant in temperate,
tropical regions were the temperature is high or low (35co -75co).
2.5 Uses and Economic Importance of Cassia occidentalis:
i. Cassia occidentalis is used as a coffee substitute and has a maturing and anodyne
action.
ii. The alcoholic or vigenar maceration of poundfresh leaves is used externally to treat
Eczema and dermatomycosis
iii. Cassia occidentalis it is useful in treating skin diseases like ringworm and itching or
body scratchand psoriasis.
iv. Decoction of the fruits of Cassia occidentalisis used in treating fever.
v. Cassia occidentalis acts as a liver stimulant, Mild laxative and heart tonic.
8
 vi. The herb helps the body in maintaining the normal level of cholesterol.
vii. It is also consumed in worm infestation and the infection occurring in the body.
viii. Its paste is used for treating skin ailment and also for getting the rid of chronic
diseases
ix. The leaves and seeds, are useful in leprosy, flatulence, colic, dyspepsia, Constipation,
cough, bronchitis and cardiac disorders.
x. The juice extracted from the cassia occidentalis leaves are useful in the case of skin
ailment, rashes, and allergic.
xi. It is used in an antidote in case of various poisonings.
xii. Its powdered proves useful in combating indigestion, Toning up heart muscles and
purifying blood.
xiii. It is worthwhile in treating piles and hemorrhoids as well as relieving the pain caused
on excretion.
2.5.1 Chemicals constitentof Casssia tora:
(+)-rhein, aloe-emodin, chrysophanol,7% resine, cathatrine, calcium, iron, phosphorus, 1,3,5,
trihydroxy-6-7—dimethoxy-2-methyla-throquinone, beta-sitosterol, naptho-alpha-phyrone-
toralactune, chrysophanol, physcion, emodine rubrofusarin, cchrysophonic acid-9-anthrone,
tricontan-1-01, stigmasterol, palmitic, stearic, succinic and d-tartaric acid, aridine, quercitrin,
isoquercctrin.
2.6 TRADITIONAL USES
Leaves were used externally and internally in skin diseases, itches, scabies and ringworm.
Hot decoction preferred to quinine for its tonic properties. Paste of leaves and calcium
hydroxide applied to abscesses for quick opening and clearing of pus. The paste of leaves was
externally applied to wounds, sores, itch, cutaneous diseases, bone fracture, fever, ringworm,
skin diseases, throat infection and to cure sore eyes. It was also used in hematuria, in
rheumatism, for typhoid fever and tuberculosis, asthma, haemoglobin disorders, to cure
leprosy and to treat diabetes. The leaves/roots were an ingredient of many popular herbal
liver tonics and medicines for liver disorders. People use it also for the treatment of insect
bites, snakebite, scorpion sting, constipation, oedema, fever, inflammation and rheumatism.
Its roots, leaves, flowers and seeds were used as laxative and purgative. The plant was also
used as febrifuge, vermifuge, anticonvulsant and against chicken pox, guinea worm and black
quarter. It was said that the infusion of the leaves of C. occidentalis is used as an effective
treatment for hepatitis. In Unani medicine it was used as an antidote of poisons, blood
purifier, expectorant, anti-inflammatory agent and a remedy for the treatment of liver
9
diseases. In Nigeria it was used for haematuria, rheumatism, typhoid, asthma and disorders of
haemoglobin (Kolhapure and Mitra, 2004).
 It is a traditional remedy for ailments such as typhoid, malaria, liver complaints, snake
bites and dog bite.
 It is also used to treat diabetes, pains, rheumatism, venereal diseases, fevers, hematuria
and convulsion.
 It is used in Ayurvedic traditional medicine in Nigeria, being the source of the drug
Kasamarda (Jain et al., 2018).
 The whole plant is diuretic, febrifuge, stomachic and tonic.
 It is used in the treatment of hypertension, biliousness, ringworm and eczema.
 Plant is boiled and gargled for treating throat troubles.
 Externally, it is pounded and mixed with wood-ash and rubbed on areas of
leishmaniasis and eczema (Basri and Fan, 2005).
 The root is cholagogue, emetic and purgative.
 An infusion is used in the treatment of bilious fever, ordinary fever, stomach-ache, and
to ease menstruation.
 Tincture of the root is rubbed onto rheumatic areas.
 Tea made from roots and dried flowers is used as a treatment for colds and upset
stomach.
 Leaf is used as a remedy for renal calculi.
 Leaves are made into a tea for treating afterbirth problems, fevers, coughs and colds,
headaches, hemorrhage and thrush.
 An ointment prepared from the leaves is applied as a remedy for ringworm and other
affections of the skin.
 Flowers are used in a preparation to reduce stomach acid in children.
 The seed is febrifuge and sedative.
 An infusion is drunk to calm one’s nerves, and as a treatment for kidney problems,
hemorrhage, worms, and cleaning womb and tubes.
 Extracts of the leaf and seed have shown antibiotic activity (Lynch and Berry, 2007).
2.6.1 USES IN MODERN MEDICINE
Various biological activities and medicinal properties of C. occidentalis is widely consumed
by the local people as a substitute for coffee. It is an ingredient in Himoliv, a polyherbal
Ayurvedic formulation. It is also proved that it prevents the carbontetrachlorideinduced
hepatotoxicity in rats. Based on the observation they suggested that Himoliv increases the
10
protective enzymes superoxide dismutase (SOD) and catalase in liver homogenate of rats. It
is also present in other polyherbal formulation Liv.52 tablet and syrup which are extensively
used for Hepatitis A (Mohammed et al., 2012).
The whole plant is diuretic, febrifuge, stomachic and tonic
It is used in the treatment of hypertension, dropsy, diabetes, fevers, biliousness, rheumatism,
ringworm and eczema
 The plant is boiled and gargled for treating throat troubles
Applied externally, it is pounded and mixed with wood-ash and rubbed on areas of
leishmaniasis and eczema.
 The root is cholagogue, emetic and purgative.
An infusion is used in the treatment of bilious fever, ordinary fever, stomach-ache, and to
ease menstruation.
 A tincture of the root is rubbed onto rheumatic areas.
A tea made from roots and dried flowers is used as a treatment for colds and upset stomach.
 The leaf is used as a remedy for renal calculi.
Leaves are made into a tea for treating afterbirth problems, fevers, coughs and colds,
headaches, haemorrhage and thrush.
An ointment prepared from the leaves is applied as a remedy for ringworm and other
affections of the skin.
The flowers are used in a preparation to reduce stomach acid in children.
 The seed is febrifuge and sedative
An infusion is drunk to calm one's nerves, and as a treatment for kidney problems,
haemorrhage, worms, and cleaning womb and tubes (Lynch and Berry, 2007).
2.7 Pharmacological Use
There are various pharmacological applications, which have been experimentally examined
for Senna. tora.
2.7.1 Anti-Oxidant Activity
Several diseases/disorders are associated to oxidative stress caused by free radicals
(Gutteridge, 2019; Mukherjee and Gogoi, 2011). Antioxidants behave as a key defense
system against free radical mediated toxicity by protecting the damages (Lee et al., 2003).
Anti-oxidants can act as free radical scavengers, lipid peroxidation inhibitor and savior to
other free radical mediated processes, protecting the human organs against several
pathologies such as Parkinson’s disease, atherosclerosis, Alzheimer’s disease and cancer
(Mates et al., 2019; Rosenkranz, 2002; Uttara et al., 2009; Rejiya et al., 2009). Scalbert et al.
11
(2005) was suggested that polyphenols may protect cell ingredients against oxidative damage
and, by that mean they limit the risk of various degenerative diseases associated with
oxidative stress. The polyphenolic content of C. tora is high (3.7 g kg-1) in dried leaves.
Keeping the fact of rich polyphenolic content in dried leaves of C. tora, in mind, Rejiya et al.
(2009) evaluated the nitric oxide scavenging activity of methanolic leaves extract of C. tora
and reducing power assays using Rutin and BHT (butylhydroxytoluene) as standards. The
extract was also studied for its lipid peroxidation inhibition assay using rat liver and brain.
Methanolic leaves extract of C. tora showed better nitric oxide scavenging activity when
compared to Rutin, so it can be used to minimize or retard the damage from nitric oxide
radicals. The methanolic leaves extract of C. tora is very effective in inhibiting lipid
peroxidation also (Rejiya et al., 2009).
2.7.2 Anti-Inflammatory Activity
The Methanolic extract of the leaves of C. tora showed good activity against carageenin,
serotonin, histamine and dextran induced rat hind paw oedema in a dose dependent manner
(Maity et al., 2018; Jain and Patil, 2010).
2.7.3 Anti-Proliferative Activity
Rejiya et al. (2009) explored the anti-proliferative potential of C. tora methanolic extract of
leaves with Cisplatin, anticancer drug in human cervical cancer cells (HeLa). This study
confirmed that C. tora methanolic extract strongly inhibited the growth of human cervical
cancer cells (Rejiya et al., 2009).
2.7.4 Hypolipidemic Activity
Ethanolic extract of seeds of C. tora and its fractions were examined for hypolipidemic
activity on triton induced hyperlipidemic profile in Albino rats. Ethanolic extract and its ether
soluble and water soluble fraction decreased the serum level of total cholesterol, LDL-
cholesterol and triglyceride while slightly increased the HDL-cholesterol level (Patil et al.,
2004; Meena et al., 2010). Cho et al. (2005) supplemented, a mixture of C. tora fiber
consisting of 2 g, 200 mg of alpha-tocopherol, 500 mg of ascorbic acid and 300 mg of
maltodextrin to type II diabetic patients for 2 months. They observed that the level of serum
total cholesterol, triglycerides and LDLcholesterol declined in the C. tora group compared
with the age and gender matched placebo group, while the Fasting blood glucose, HbA1c,
blood urea, creatinine and activities of serum Aspartate Aminotransferase (AST) and alanine
Aminotransferase (ALT) were not altered (Cho et al., 2005).

12
2.7.5 Anti-Diabetic Activity
The hypoglycemic activity of C. tora has been reported by many scientists. Nam and Choi
(2008) studied the effects of C. tora L. seed butanol fraction (CATO) on postprandial glucose
control and insulin secretion from the pancreas of the normal and streptozotocin induced
diabetic
rats. They observed that in normal rats fed with CATO have lower postprandial glucose
levels. In diabetic rats, the levels in the CATO fed group have lower postprandial glucose
during the 30~180 min.
2.8 Cowpea weevil (Callosobruchus maculatus)
2.8.1 Description
Cowpea weevil (Callosobruchus maculatus) (Coleoptera: Bruchidae) is an invasive species
of beetle that attacks stored legumes (Fabaceae). Callosobruchus maculatus is distributed
throughout the tropical and subtropical regions of the world but it is dominant in Africa.
Cowpea weevil belongs to the genus Callosobruchus in the subfamily Bruchinae and family
Chrysomelidae (Kergoat et al., 2017). Callosobruchus maculatus adults are 2.0-3.5 mm long
and lack the snout of a true weevil. It is reddishbrown overall, with black and gray elytra
marked with two central black spots. Two forms of this species exist; a dispersal (flying)
form and a sedentary (flightless) form. The sex difference in flightless form is quite
distinctive while the sexes in flying form are subtly dimorphic (Beck and Blumer, 2018).
Figure 2.2 show male and female cowpea weevil.

Figure 2.2 Cowpea weevil (♂: male, ♀: female) The images are of the same scale and the
squares are 1mm. Source: Beck and Blumer, (2018).
2.8.2 Life cycle
Upon infestation, the weevil lays a single fertilized egg on the surface of the grain. The egg is
characteristically spindle-shaped, clear and shiny. It develops into a larva that burrows into
the endosperm and attacks the grains from within. The developing larva of the weevil feeds

13
entirely on the grains. After a series of molting, the larva is ready for pupation. It burrows to
a location beneath the seed coat where it emerges as an adult insect. The emergence of the
weevil leaves a hole in the grain. The adult weevil attains maturity 24 to 36 hours after
emergence. The adult weevil has a lifespan of about 1 to 2 weeks. Adulthood of C. maculatus
is wholly devoted to mating and egg-laying (Beck and Blumer, 2018). Under favorable
conditions, the whole metamorphosis occurs within 45- 48 days (Devi, 2018).
Figure 2.3 shows different developmental stage of cowpea weevil during metamorphosis.
2.9 Storage pests control strategies
2.9.1 Cultural and sanitary control
Cultural and sanitary strategies of insect pest control involve specific practices implemented
to reduce the likelihood of insect infestation and damage. Sanitary control techniques entail
crop hygiene procedures such as cleaning of storage vessel to remove debris, laid eggs, larvae
and dead insects left after consumption of the grain. It also involves aeration of granaries
before the next batch of grain is brought in for storage (Newman, 2018). Cultural method of
pest control involves cultivation of resistant crop varieties, crop rotation and/or the use of trap
crops to keep the pest away from the desirable crop (Beck and Blumer, 2018).
2.9.2 Physical control
Physical control of insect pests forms an integral part of IPM. This method plays a role in
pest prevention, monitoring and control (Alder, 2020). To prevent an infestation, a uniform
temperature above 55°C is applied to the grains for 60 minutes or 60°C for about one minute.
Temperatures can also be lowered below 13°C to prevent insect development. Grains can also
be stored in hermetic storage structures or packed in insect-proof packages (Riudavets et al.,
2017).
For the control of pests, extreme temperatures and mechanical methods are employed. The
process involves the application of vacuum to the stored products in order to eliminate
oxygen from the inter-granular spaces. Carbon dioxide can also be applied in pressure
chambers at high pressures of up to 35 bars for few hours. In addition, high temperatures
above 50°C can be applied to the structure. The stored product can also be cold treated at
temperatures of about -20°C for 24 hours (Adler, 2016). The advantage of using physical
method of insect control is that the insects do not develop resistance. The quality of the
product is not altered and is free from toxic residues (Berger, 2018; Sharma et al., 2016).
2.9.3 Biological control
Biological control is an effective tool in pest management either at the fields or at the storage
level. It involves strategies such as the use of cultivars resistant to pest, pheromone traps and
14
the use of natural enemies to control or suppress insect pests (Hodges, 2018). Most biological
agents used against insects include parasitoids, predators and pathogens (Futch, 2020). The
use of parasitoids has been commercially exploited for the control of pyralid moths and
beetles of stored products (Schöller et al., 2016). The parasitoids used in the control of
cowpea weevils are the larval parasitoids Dirnamus basalus and Eupelmus ssp and the egg
parasitoid Uscana lariophaga (Stolk et al., 2018). The use of biological method possesses a
number of advantages. It does not pose health and environmental hazards. The use of natural
predators is specific to the target pest and do not harm the beneficial organisms. The demerits
associated with biological control of pests is that it is a costly method of pest control. It rarely
eliminates pest and therefore not the ideal method for the control of heavy infested grains
(Scholler, 2020).
2.9.4 Chemical control
Chemical control of insect pests entails prevention and eradication of pests using insecticides.
Insecticides could be synthetic or naturally occurring chemicals that kill or alter physiological
functions of insects. These chemicals vary in chemical composition and structural
arrangement of compounds. Commonly used classes of pesticides include organophosphates,
organochlorides, carbamates and pyrethroids (Schöller et al., 2016).
Insecticides use different routes of entry into the target site in the insect. Stomach poisons are
ingested with food, contacts insecticides are absorbed via the cuticle and fumigants enter via
the spiracles and tracheae. Insecticides employ different modes of action in the insect. They
can be neuroactive agents, insect growth regulators, metabolic inhibitors and physical
poisons. Organophosphate and carbamates are synaptic poisons that target the
acetylcholinesterase (AChE). They inhibit AChE by phosphorylation (organophosphates) and
carbomylation (carbamates). This inhibition results in accumulation of acetylcholine at the
synaptic junctions. This causes tremors, uncoordinated movements, paralysis and eventual
death of the insect (Das, 2013). Pyrethroids, the derivatives of pyrethrin, are axonal poisons.
They target the voltage-gated sodium ion channels. These sodium channel modulators
prolong the action potential of the nerves. Juvenile Hormone Analogue such as the hydropene
and methopene; fenoxycarb and Pyriproxyfen mimic the juvenile hormone in the insects
(Schöller et al., 2016).
The use of synthetic chemicals is a popular method of pests control because they are
effective, fast-acting and easy to use as compared with other control options (Futch, 2020).
This method, however, is bedeviled by a number of economic and ecological shortcomings.
These include residual toxicity, adverse effects on the beneficial and nontarget organisms, the
15
risk of user’s contamination, pesticide resistance and high-cost of purchase (El-Kamali,
2019). Aldrin and dieldrin caused the death of seed-eating game birds, raptors that preyed on
the game birds and also the death of fish-eating birds (Gay, 2021). Pirimiphos-methyl
(Actellic), an organophosphate pesticide, has been established to affect male rat reproductive
performances (Ngoula et al., 2017). The IPM program directs the use of chemical pesticides
as the last resort in the control of pest; and it should be specific to the target (DAERA, 2016).
2.10 Insecticidal properties of plants
Plants are a rich source of bioactive compounds including the alkaloids, terpenoids, steroids,
carotenoids, flavanoids, glycosides and a range of essential oils. These compounds are
products of secondary metabolism in plants. They do not serve any physiological functions in
plants and neither are they present in all plant species. Phytochemicals serve a defense
function in plants. Their evolution is attributed to selective pressure exerted by plants in a
process of self-defense against pests (Arnason et al., 2019).
The existing literature indicates that these phytochemical constituents contain an array of
properties including insecticidal, antioxidant, anti-diabetic and antibacterial among others.
Phytochemicals are produced in larger quantities during pest invasion. They have varied
effects on insects. They can kill the insect, repel or attract, alter their development and even
affect oviposition (Chowański et al., 2016). Various mechanisms of action are employed by
these secondary metabolites. Some act on the nervous system. They target the
acetylcholinesterase enzyme, the acetylcholine receptors, the gamma-aminobutyric acid
receptors and the voltage-gated sodium ion channels. Some acts as growth hormone
regulators (IGRs). They mimic the Juvenile hormone and ecdysone hormone in the insects
altering their growth and development (Silva, 2018). The biocidal activity of phytochemicals
occurs when they target the central nervous system (Silver et al., 2018).
Phytochemicals may also disrupt the basic metabolic functions of the cells. Utilization of
secondary metabolites as plant protectant dates back to about 400 BC during the ancient
Rome (Dayan et al., 2019). In the 17th century, nicotine derived from tobacco leaves was
used to protect plum from beetles (Silva, 2018). Other botanicals that have been employed in
the management of pest throughout history include sabadilla, rotenone, pyrethrum and neem
(Grdisa and Grsic, 2016). Neem has been intensively exploited and is the only plant from
which several commercial products have been developed (Obeng-Ofori, 2010).

16
 CHAPTER THREE
3.0 MATERIALS AND METHODS
3.1 Equipment
Equipment Manufacturer Model No./quality/spesification
Aluminium Foil Bags Nigeria Aluminium Foil Bags
Mortar and Pestle Humboldt Mfg. Co. Model No. 7563
Weighing balance S.V Gupta Single-pan mechanical Balance
Filter paper BBLTM P220-B Grit Psa Paper roll
Petri dishes England W02-petri
Test tube SCM. Chaina Pyrex glass
Shovel
Soil
Beaker Pyrex glass
Measuring cylinder Pyrex glass
Conical flask Pyrex glass
Spatula
Test tube rack Pyrex glass
Syringe WW.AG-17024 Jangxi jin sheng/china
Heating mantel

3.1.2 Reagents

17
 Reagent Purity Specification Company
Distilled water 96%
Sulfuric acid (H2SO4) 98% E Merck
Acetic acid 98% E Merck
Hydrochloric acid (HCL) 97% E Merck
Methanolic 98% E Merck
Ferric chloride 85% E Merck

3.2 Sample collection

18
The sample was obtained from Ngaski Local Government Kebbi State, Birnin Kebbi Nigeria.
In a sterile polythene bags, this leaves were transported to the Department of Science
Technology, Waziri Umaru Federal Polytchnic and where kept in Biochemistry Laboratory
for further analysis.
3.3 Preparation of extracts
The plant material were chopped into bits and air dried for 2 weeks. And was then grinded in
to powder with micro plant grinding machine and was sieved through a 0.25mm pore size to
obtain a uniform fine dust particle (Hudaib et al., 2016). The resulting powder was stored in
separate containers with screw cap at room temperature prior to use. The amount of powder
mixed with the 100ml of water was calculated on weight by volume, ie: weight of powder
/volume of water 10 , 20, 30g of each grinded plant material will be soaked in 100ml of water
to obtain crude extracts of three concentration levels of 10, 20 and 30%(w/v).Each mixture
will be filtered with cheese cloth after 24 hours.
3.4 INSECT CULTURE
The insect pest used in this study was Cowpae weevil, cassia occidentalis. The weevil were
reared on cawpea seeds (L.) walp in the laboratory to adapt to the laboratory condition using
the method described by Hudaib et al., (2013).The spinach used as substrate to culture the
weevil were thoroughly cleaned and exposed in an oven to ensure the absence of Insect, mites
and disease-causing microorganisms. Batches of ten (10)treated samples were placed in five
different plastic containers previously washed, sterilized and dried. Five (5)weevils isolated
from the infested spinach samples were introduced in to each plastic container and covered
with polythene net fastened tightly with rubber band. The spinach-weevil mixture were kept
in the laboratory for 4 days to allow mating and oviposition to occur after which the parents
weevils were removed. The rearing was given sufficient time (2-5 days) until new adult
Insect emerged. The first finial generations (F1) adult weevils used for the experiment were
1-2 days old after emergence.
3.5 TOXICITY BIOASSAY OF EXTRACTS
The plant extracts were assayed for insecticidal potency using the method described by lwu,
(2013). Sample previously disinfected were divided into three lots of 10g each and replicated
thrice. Each set of sample was placed in a test tube (14.7×2.4 cm) and treated with the plant
extracts of different concentrations (50.0, 75.0 and 100.0 mg/mL). The tube were manually
rocked for two minutes to ensure that the sample were coated with the extracts after which
they were removed from the tube and placed on filter papers for 24 h to allow the solvent to
evaporate. Then each lot of seeds was placed in separate fresh test tubes and ten (10)adult.
19
3.6 PHYTOCHEMICAL SCREENING (QUALITATIVE TEST)
Phytochemical analysis of the cassia occidentalis extract were carried out in accordance to
the method described by (Hudaib et al., 2016)
3.6.1 Test for quinones :
1ml of extract + few ml of alcoholic potlussium hydroxydes red or blue colour shows the
presence of quinones
3.6.2 Test for authraquinones:
10ml of 10%NH3 solution + few ml of filtrate ( shaven vigoroush for 30sec). A pink, violet
or red colour indicate the presence of authraquinonots
3.6.3 Test for cardiac glycosides:
1ml of filtrate + 1 5ml glacial acetic acid + 1 drop of 5% ferric chloride + conic H2504. Blue
colored solution ( in acetic layer) indicate the presence of cardiac glycoside.
3.6.4 Test for taunins:
1ml extract + 3 ml distilled water +3 drops of 10% ferric chloride solution. Appearance of
blue-green colour indicate the presence of Tannins.
3.6.5 Test for phenolic compound :
1ml of extract + few drops of diluted hydrocloric acid. A transient red- coloured show the
presence of phenolic compound.
3.6.6 Test for terpinoides :
2 ml chloroform + 5ml plant extract ( evaporated in water bath) +3 ml conc. H2504 (boil in
water bath). A grey coloured solution show the presence of terpinoides.
3.6.7 Test for anthocyanin:
2 ml extract +2 ml2N Hcl+ few ml ammonia . pink- red solution which turns blue violet after
addition of ammonia shows the presence of Anthocyonins.
3.6.8 Test for volatile oil :
2 ml extract + shaken with 0.1ml Na0H + few drops of dil. Hcl. White pp+ indicate the
presence of volatile oil.
3.6.9 Test for carboxylic acid :
1 ml extract +1ml sodium bicarbonate solution appearance of efffervence shows the presence
of carboxylic acid.

3.6.10 Test for alkaloids :

20
3ml of extract + few drops of iodine solution a blue colour, disppear on boiling and reappear
on cooling shows the presence of alkaloid
OR
Few ml extract +3-4 drops of 2%picnic acid solutions. An orange colour show the presence
of alkaloid.
3.6.11 Test for phytosterols:
2 ml of fitrate + few drops of conc. H2504 ( shaken well and allow to stand) appear of red
color in lower layer indicate the presence of phytosterols.
3.6.12 Test for flavonoides
1ml extract +2ml 2%Na0H solution + few drops of dil. Hcl An intensive yellow colour,
become, colorless in addition of dil acid.

21
 CHAPTER FOUR

RESULT, DISCUSSION, CONCLUSION AND RECOMMENDATIO

4.1 RESULT

Table 4.1 Result of quantitative phytochemical analysis of Guiera senegalensis

S/N Parameter Result

1. Quinone N.D
2. Anthraquinone N.D
3. Cardiac Glycoside ++
4. Taunins +++
5. Phenolic ++
6. Terpinoides Boil
7. Anthocyanin +
8. Volatile Oil N.D
9. Carboxylic Acid N.D
10. Alkanoids +
11. Phytosterols X
12. Flavonoids +

22
4.2 Discussion

The phytoconstitutent research on Cassia occidentalis showed the presence of diverse groups
of boiactive constituent whose biological activity can be valuable therapeutic index. The
result of the present phytoconstitutent showed the presence of flavonoid, saponin, while
tannin, alkaloid.
Flavonoids are now recognized as possessing an array of bioactivities with several
mechanisms relevant to potential reductions in the pathogenesis of chronic diseases (e.g.,
anti-inflammatory and antioxidant actions as well as alteration of redox-sensitive signal
transduction pathways and gene expression) (Ajayi et al., 2018).
Tannins were slightly present. Generally Tannins have different functions in that they serve
as chelating agents for metals ion, antioxidants in biological systems, and as protein
precipitating agents (Asokan and Vanitha, 2018).
Saponins were largely present in the seed oil sample. Because of their surfactant nature, they
are used industrially in mining and ore separation, in preparation of emulsions for
photographic films, and, extensively, in cosmetics, such as cleansing formulae. In addition to
their emollient effects, the antifungal and antibacterial properties of saponins are important in
cosmetic applications (Cook et al., 2015).
Alkaloids figure as a very prominent class of defense compounds. Alkaloids were found
largely present in Parinarimacrophylla seed oil extract which signifies their functionality in
repellence, deterrence, toxicity and growth inhibition by herbivores/ predators and in growth
inhibition and toxicity by microbes/ viruses and as secondary metabolites for UV- Protection
and Nitrogen storage.

23
4.3 CONCLUSION
In conclusion base on the research Cassia occidentalis L. is an annual or perennial plant
which is used in several traditional medicines to cure various diseases. C. occidentalis is a
spiny herb grows all over in Nigeria in shade as well as under open condition. C.
occidentalisis a perennial plant which is erect and annual. The plant grows up to 2m tall. The
annual herb is soft wooded, foetid. The stems are greenish brown in colour and are smooth in
texture. The primary roots of C. occidentalis are diarch in nature and they have a
dicotyledonous secondary growth.

24
4.4 Recommendation
It is recommended that more research to be conducted on cassia occidentalis rind in order to
establish the presence of other phytochemicals and also to quantify them.

25
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