Chapter 2

Literature Review
Mosquitoes
Mosquitoes belong to family Culicidae of the order Diptera and are similar in appearance
to other flies. They pass through four distinct stages in their life cycle: egg, larva, pupa, and
adult. Prodigious numbers of mosquitoes can hatch simultaneously under the proper conditions.
Eggs are white in color when first deposited but darken within 12 to 24 hours. Eggs of different
species vary from canoe-shaped to elongate-oval in, laid singly or glued together forming rafts.
The incubation period is dependent on environmental and genetic factors and varies considerably
among different species. Mosquitos deposit their eggs directly on the water surface (some in
crevices of soil, or on special substrates or niches that are or will subsequently be flooded) and
may hatch in one to four days depending on temperature. Larvae (wigglers or wrigglers) of all
mosquito species live in the water and most are filter feeders. Depending on the species and
environmental conditions, mosquitoes may take three to four days up to several weeks to
complete larval development. Mature 4th instar larvae then molt to the pupal stage. Pupae can be
very active and are often called tumblers due to their rapid, tumblinglike movement when
disturbed. They do not feed, they breathe through two respiratory horns when at the surface
and typically transform into adults in two or more days. Adult mosquitoes are terrestrial and
capable of flight. Females feed mostly on animal blood with their piercing-sucking mouthparts,
while males mouthparts are modified where no piercing is required (Public-Health Pesticide
Applicator Training Manual) FIND SPECIFIC SOURCE !!!
Mosquitoes are capable of transmitting several vector-borne diseases than any other
groups of arthropods and can affect millions of people. According to Massebo 2009, Mosquitoborne diseases contribute significantly to disease burden, death, poverty, and social debility all
over the world, particularly in tropical countries. Among these diseases, dengue remains the most
serious vector- borne disease in the Philippines. Dengue is a severe, often fatal, most rapidly
emerging febrile disease transmitted by female Aedes aegypti mosquitoes. It is an acute infection
that kills much faster than AIDS. In the Philippines, dengue fever and dengue hemorrhagic fever
are widespread in all its regions according to World Health Organization.
According to a research by Ghosh et al. 2012, many institutions and research bodies are
looking for solutions in controlling the vector of dengue because no specific therapy is still
available to date. Most of the measures and proven effective way available includes the use of
commercialized products containing chemicals in order to control vectors. However, synthetic
controls may cause resistance and resurgence of mosquito populations, poisoning to humans and
livestock, and hazardous to the environment. The reason for extensive use of plants as drugs may
be the fact that plants are available everywhere, wide range of medicinal plants and their distinct
form and thus are procured without any trouble. Medicinal plants have a long history of use and
their use is widespread in both developing and under developed countries (Kinghorn, 2010). This
aspect initiated the research for alternative measures that are environment-friendly and
degradable. Plant-based control methods are now being deliberated because plants consist of
several bioactive components that are of low toxicity and biodegradable. Unlike the conventional
insecticide that is based on single active ingredient, green insecticides contains botanical
combinations of bioactive chemical compounds which act concertedly on both behavioral and
physiological processes.

Banana peel utilization

There is an urgent need to seek resource and value-added use for fruit wastes as it
becomes one of the main sources of municipal solid wastes in many countries. In fact,
inexpensive and readily available use of agri-food industry waste is highly cost-effective and
minimizes environmental impact. One of the most beneficial approaches is to recover the
bioactive constituents, especially the phenolic compounds making full use of them in the food,
pharmaceutical as well as cosmetics industry. Phenolic compounds are one of the most important
categories of natural antioxidants of interest. In recent studies, the antioxidant potency and the
content of phenolic compounds were found to be high in the peel and seed of some fruits
indicating that fruit residues have the potential to be utilized as a source of bioactive compounds,
such as natural antioxidants (Deng, 2012)
The Department of Agriculture states that banana (Musa) is the leading fruit grown in the
Philippines and is a consistent top dollar earner. It is the second largest produced fruit crop in the
world after citrus and the Philippines ranked fifth in banana producing nation. Musa is genus
from Zingiberales and family Musaceae; it includes bananas and plantains. There are around 70
species of Musa with a broad variety of uses. The common was banana scientifically known as
Musa sapientum (Chabuck, 2013)
Several studies have proven that banana plant parts are useful in many ways, such as
antioxidant, anti-inflammatory, antimicrobial, and as an insecticide.
According to a study made by Wuyts, et. al, there are many composition of banana skin
like enzymes such as polyphenoloxidase, pectin as gelling agent and that the banana peel extract
is used alone or combined with a cream or ointment, medicinal benefits of the extract include
relief of pain, swelling and itching.
According to Emaga et. al, banana peel is a rich source of starch (3%), crude protein (69%), crude fat (3.8-11%), total dietary fiber (43.2-49.7%), and polyunsaturated fatty acids,
particularly linoleic acid and -linoleic acid, pectin, essential amino acids (leucine, valine,
phenylalanine, and threonine), and micronutrients (K, P, Ca, Mg).

Phenolic compounds

Bioactive components of plants as larvicidal/ insecticidal agent

The use of different parts of locally available plants and their various products in the
control of mosquitoes has been well established globally by numerous researchers. The larvicidal
properties of plants have also been documented in many countries along with the repellent and
anti-juvenile hormones activities (Singh and Bansal, 2003). Traditionally, plants and their
derivatives were used to kill mosquitoes and other household and agricultural pests. In all
probability, these plants used to control insects contained insecticidal phytochemicals that were
predominantly secondary compounds produced by plants to protect themselves against
herbivorous insects (Shaalan et. al, 2005; Precti Sharma et. al, 2009).

Hamouda et.al (2015) conducted a study utilizing pomegranate fruit peel extract as
insecticide against two pests, Myzus persicae (peach-potato aphid) and Phthorimae operculella
(potato tuber moth), and antifungal to four fungi, Botrytis cinerea, Fusarium sambucinum,
Penicillium digitatum and Aspergillus niger. Subjects were treated with aqueous, ethanol, and
methanol extracts used. Results showed that all extracts of the fruit peel that were examined had
a significantly high toxicity effect on Myzus persicae as compared to untreated control.
Ethanolic, methanolic and aqueous peel fruit extracts of pomegranate had a high preventive
effect on potato tuber moth larval penetration with 77% less than the untreated control.
Application of all extracts of the fruit peel on potato tubers reduced egg-laying of P. operculella
with an average number of 4.8 (ethanol), 6.6 (methanol), 8.4 (aqueous). These results
demonstrated that pomegranate extracts had inhibitory effects against the pest. Koide et.al
reported that toxicity caused by the fruit is due to the astringent properties of tannins contained in
the peel fruit which stop insects infestation. Moilanen and Salminen showed that peel is rich in
ellagitannins that are considered as toxic against insects.
Another study made by Hamouda et.al reveals that phytochemicals or crude plant extracts
of pomegranate peel have insecticidal activity against Tribolium castaneum (red flour beetle). T.
castaneum is one of the most severe secondary insect pests that feeds on a widespread range of
long-lasting stored grain products. The results showed insecticidal effects of the three extracts,
aqueous, ethanol, and methanol, of pomegranate fruit peel. However, ethanol was found to be the
most toxic having the highest mortality rate on larvae and adult beetles. Toxicity of ethanol
extract on T. castaneum was expressed by 56% larval mortality after ingestion. In this respect,
Koide et.al reported that toxicity caused by pomegranate fruit is also due to tannin content of the
peel that has astringent properties.
Aloe vera is a perennial plant belonging to the family of Liliaceae. Experiments using the
leaf extract exhibited 34% mortality of 1st instar larvae of A. aegypti at 80 ppm, and 89% when
increased to 400 ppm (Subramaniam, 2012).
Torres et. al (2014) evaluated the larvicidal toxicity of ethanol and hexane leaf extracts of
Annona muricata L. (graviola/ soursop) collected from different localities in the Philippines
against 3rd and 4th instars larvae of A. aegypti following the World Health Organization bioassay
method. The mortality was observed 24 and 48 hours after treatment and data probit analysis was
conducted to determine lethal concentrations (LC50 and LC90). The extract was characterized by
phytochemical analysis. The results revealed that both hexane and ethanol extracts exhibited
evidence of strong larvicidal toxicity. The hexane extract of the leaves showed the highest
toxicity with LC50 value of 25.99 mg/L and LC90 value of 49.47 mg/L while the ethanol extract
yielded an LC50 of 46.36 mg/L and LC90 of 195.39 mg/L. Characterization of the ethanol
extract showed presence of flavonoids (leucoanthocyanins), condensed tannins, unsaturated
steroids and triterpenoids and fats and oils.
A study of the flowering plant, sage, Lantana camara belonging to the Family
Verbanaceae, of Kumar and Maneemegalai, evaluates its larvicidal effect against A. aegypti and
Culex quinquefasciatus. Larvicidal effect on 3rd and 4th instar larvae of mosquito species A.
aegypti and C. quinquefasciatus have been investigated in a dose dependent manner for 24 hours.
With 1.0 mg/ml concentration of extracts of Lantana camara, maximum mortality was observed
in A. aegypti exposed for 24 hours. In the case of C. quinquefasciatus the mortality was seen
maximized when the concentration increased to 3.0mg/ml. Presence of saponin, flavonoids,

terpenoids and cardiac glycosides have also been observed and GC/MS analysis was carried out
on methanol flower and leaf extract to find out the components.
A study in the efficacy of the phytochemical saponin in the fruit extract of Balanites
aegyptiaca (desert date) against third instar larvae of Aedes aegypti mosquito was made by
Chapagain & Weisman (2005). Treatment composed of methanolic extract of mesocarp (MCE),
crude ssaponin extract (CSE), and pure saponin fraction (PSF). The results showed that all three
extracts used in the experiment were found active against the larvae. However, PSF was found to
be most active with regard to its efficacy to inhibit adult emergence. Exposure of the early fourth
instars A. aegypti larvae to 35 ppm prevented the emergence of 50% of the treated populations,
whereas a concentration of 260 ppm and 85 ppm of each saponin extract (CSE) and methanolic
extract (MCE) respectively was needed to inhibit 50% of the tested population. The results in the
study clearly show that saponin fraction of the B. aegyptiaca fruit mesocarp is highly larvicidal
against the A. aegypti mosquito larvae compare to crude saponin and methanolic extracts.
Anupam Ghosh, et. al 2012, listed in their study the various plant extracts that has a
potential use as mosquito larvicide. The study summaries the application of phytochemicals
found in plants, the nature of active ingredients responsible for larval toxicity, and the mode of
action of phytochemicals in target insect body.
High tannin concentrations are found in nearly every part of the plant, such as in the bark,
wood, leaves, fruit, roots, and seed. Frequently an increased tannin production can be associated
with some sickness of the plant. Therefore, it is assumed that the biological role in the plant of
many tannins is related to protection against infection, insects, or animal herbivory. The tannins
appear as light yellow or white amorphous powders or shiny, nearly colorless, loose masses, with
a characteristic strange smell and astringent taste (Khanbabaee 2001; Haslam; Porter 1989).

Bioactive components of Musa as larvicidal/ insecticidal agent

According to the study made by Maniyan et. al., fruit peels are an important source of
bioactive compounds including antioxidants and proteins. Phytochemicals are diverse range of
biologically active compounds found in plants, which provide color, flavor and natural protection
against pests to the plants. These compounds play a role in reducing the risk of developing many
forms of cancer (lung, prostate, etc.) and risk of cardiovascular disease. Additional important
bioactive components present in fruit peels are tannin, phytic acid, and proteases with each
having their own significance and biological roles. Results of the study shows that banana
yielded a carbohydrate content of 32.64 mg/ml, 0.11 mg/ml of proteases, and 25 g/ml of tannin.
The astringent taste of unripe banana is due to the presence of phenolic compounds.
Bananas are rich in phenolic compounds and flavonoids, which have antioxidant properties.
Dopamine, which is an antioxidant reacts with polyphenol oxidase, monophenol monooxygenase
an o-diphenoloxidase, which produces tannins resulting in brown spots on peel (Kanazawa,
2000; Alothman, 2009). A change in the peel color is largely due to degradation of chlorophyll or
unmasking of carotenoids, rather than carotenoid synthesis.
In addition to enzymes, phlobatannins, alkaloids, glycosides and terpenoids were found to
be present in the peels of genus Musa. These phytochemicals have been reported to exert
multiple biological and pharmacological effects such as antibacterial, antihypertensive,

antidiabetic and anti- inflammatory activities. The presence of these bioactive substances in
banana peels therefore suggests that the peels possess valuable medicinal potential yet to be
explored as the bioactive compounds contained in plants are majorly responsible for their
medicinal properties (Ighodaroro, 2009).
Velumani (2016) identified banana (Musa paradaisica), as being grown worldwide and
consumed as ripe fruit or used for culinary purposes. Peels form about 18-33% of the whole fruit
and are a waste product. With a view to exploiting banana peel as a source of valuable
components, the nutritional composition, antioxidant components were determined. The
phytochemical analysis of aqueous and organic solvent extracts of banana peel reveal the
presence of alkaloids, flavanoids, carbohydrates, proteins, tannins, terpenoid, saponins,
glycosides and anthroquiones. The phytochemicals alkaloids, flavanoids and tannins have a very
good antioxidant property while saponins and terpenoids have antimicrobial activity. The
qualitative analysis of aqueous and organic solvent extracts of banana peel reveals the presence
of reducing sugar (glucose and fructose). The qualitative analysis of amino acids in banana peel
reveals the presence of tryptophan, phenylanine, tyrosine, ariginine and cysteine. The
quantitative analysis of starch, proteins were done. The proteins and carbohydrates contents were
higher in M. paradisiaca peel extracts of ethanol. The qualitative analysis of tryptophan were
carried out in M. paradisiaca peel extract and the tryptophan was found to be higher in water and
ethanol extracts.
The aqueous and soxhlet extracts of the Elaeis guineensis (palm tree) tusks, Musa
paradisiaca (plantain) peels and potassium carbonate (potash) were screened for the presence of
phytochemicals and their antimicrobial activities against the bacteria isolate from fermented
Pentaclethra macrophylla seeds were investigated by Kemka-Evans, 2013. The bacteria isolate
were Bacillus sp. and Streptococcus sp. The extract showed secondary metabolites such as
tannins, saponin, anthranoid, anthraquinone, phenol, alkaloid, phylobatannins and cardiac
glycoside. The extracts inhibited the growth of the bacteria isolates. The zones of inhibition
exhibited ranged from 7mm -8mm in M. paradisiaca peels extract, 7mm 8mm in E. guineensis
tusk extract and 7mm 8.5 in potassium carbonate extract.
Deng et.al (2012) systematically evaluated the presence of bioactive compounds on
various fruit peels. The results showed that banana peel contains 2.360.10 and 1.210.33 of fatsoluble and water soluble fraction using Ferric-Reducing Antioxidant Power (FRAP) (mol
Fe(II)/g) of peel, respectively, with a total of 3.570.43, antioxidant capacity. The bioactive
compounds quantified and identified were catechin, cyanidin 3-glucoside, epicatechin, galangin,
gallic acid, homogentisic acid, kaempferol, and chlorogenic acid.
Phenolic compounds are the most studied secondary metabolites because of their
considerable involvement in plant-environment interactions. They are molecules belonging to
very diverse chemical families having in common an aromatic ring bearing at least one phenol
hydroxyl substituent. Some phenolic compounds have several hydroxyl group substituents,
which can undergo esterification, methylation, etherification or glycosylation reactions (Raven
et. al., 2003; Macheix et. al., 2005; Lattanzio et. al., 2006). The phenolic compounds of plants
include the simplest forms (hydroxybenzoic and hydroxycinnamic acids), condensed forms
(tannins), and forms related to non-phenolic macromolecules (certain glucidic components of the

pecto-cellulosic wall, cutin and suberin). Only two groups of phenolic compounds are mainly
related to the plant walls: firstly, low- molecular-weight hydroxycinnamic acids related to
various cell wall compounds, and secondly, lignins, which are polymers of monolignol units
bound by oxidative coupling (Ewan, et. al. 2012)

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