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Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 7
.
RREEVVIIEEWW OOFF LLIITTEERRAATTUURREE
The literature pertaining to the study, “In silico approaches for characterizing
antimicrobials and larvicidal activity of selected plant extracts against the filarial vector,
Culex quinquefasciatus (Diptera: Culicidae)” is reviewed under the following headings:
2.1. Larvicidal activity of plant products against mosquitoesThe use of biologically active plant materials with anti-larval properties has
attracted considerable interest of scientists all over the world. Because of their
biodegradable nature and being relatively safer to human beings and non target organisms
in the environment, extensive survey of the flora was undertaken to search for potential
plant extracts which could be used in the management of agricultural and household pests.
Moreover, investigations on the insecticidal properties of plant extracts have gained great
impetus because of imposition of restrictions on the use of chemical pesticides for insect
control.
Saxena and Sumithra (1988) noted that the leaf extracts of Ipomoea carnea
fistulosa significantly increased the average larval and pupal development periods of the
mosquito, C. quinquefasciatus and also resulted in increased larval and pupal mortality.
Evans and Raj (1988) observed that the crushed aqueous extracts of Quassia amara,
T. neriifolia, Anacardium occidentale, Carica papaya, Hevea brasilionsis and Nerium
indicum showed larvicidal activity when tested against C. quinquefasciatus.
Thangam and Kathiresan (1988) reported that the acetone extracts of 12 sea weeds
showed larvicidal activity against the fourth instar larvae of A. stephensi and
C. quinquefasciatus. Growth inhibitory properties and toxicity of Melia volkensii fruit
extract fractions against mosquito larvae had been reported for A. arabiensis (Mwangi and
Mukiama, 1988), A. aegypti (Mwangi and Rembold, 1988) and C. quinquefasciatus
(Al-Sharbok et al., 1991).
Evans and Raj (1991) studied the larvicidal efficacy of Quassin against
C. quinquefasciatus. Crushed aqueous extracts of leaf, wood, bark and flowers of Quassia
amara showed antilarval activity against C. quinquefasciatus. Quassin has been identified
2
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 8
to be the antilarval principle present in this plant and was effective against mosquito larvae
at a concentration of 6 ppm. Jalees et al. (1993) investigated the crude ethanolic extract of
C. sativa, to determine its insecticidal properties against the larvae of An. Stephensi,
C. quinquefasciatus and Ae. aegypti.
Achary et al. (1993) evaluated the efficacy of Iopmoea carnea to control the larvae
of C. quinquefasciatus. Mwaiko and Savaeli (1994) carried out tests on lemon peel oil
extract as a mosquito larvicide. The oil was found to be toxic to larvae, pupae and eggs of
C. quinquefasciatus. Mohsen et al. (1995) found that the crude ethanolic extracts of the
weed. I. cylindrical exerted moderate larvicidal and oviposition deterring activities against
C. quinquefasciatus and produced 100% mortality against third instar larvae at 1500 ppm.
The leaf extract of Vitex negundo, Nerium oleander and seed extract of Syzigium
jambolanum exhibited larvicidal activity against C. quinquefasciatus and A. stepensi larvae
(Pushpalatha and Muthukrishnan, 1995).
Mittal et al. (1995) determined the efficacy of six neem products against fourth
instar larvae of An. stephensi, C. quinquefasciatus and Ae. aegypti. These neem products
showed poor larvicidal activity within 24 h exposure, but some proved to have good insect
growth regular activity. Larvae of An. Stephensi were generally the most susceptible to
these neem products while Ae. aegypti were the least.
Bowers et al. (1995) obtained organosoluble extracts from 55 Turkish mecidinal
plants and tested them understandardized condition for biological activity against third
instar larvae of mosquitoes, A. aegypti and A. gambiae. Eight extracts demonstrated
significant larvicidal activity with A. gambiae being more susceptible than A. aegypti in all
cases. Macedo et al. (1997) tested 83 plants in the state of Minus Gerais, Brazil for the
larvicidal activity against the mosquito A. fluviatilis. The ethanol extract of Tagetes minuta
showed active results whereas extracts of Achyryrocline satureoides, Gnaphalium
spicatum, Senecio brasiliensis, Trixis vanthieri, T. patula and Vernonia ammophila showed
less active.
Thangam and Kathiresan (1996) investigated the sea weeds, sea grasses and
mangrove plants for their larvicidal, skin and smoke repellent activities against mosquito
species. Some of them were effective in killing the larvae or repelling adult female
mosquitoes. Leaves of Excoecaria agallaha and Acanthus ilicifolius were found to show
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 9
smoke repellent activity. Isolation and identification of active compounds from the
effective samples would be useful in synthesizing mosquito larvicides or repellents on a
large scale.
Karmegam et al. (1997) investigated bioassay studies of laboratory colonized
larvae and field collected larvae of C. quinquefasciatus with 5 concentrations of petroleum
ether extracts of 12 plants collected and showed that the extracts of Argemone mexicana,
Jatropha curcus, Pergularia extensa and Withania somnifera had acute toxicity.
Batra et al. (1998) examined neem oil water emulsion was used in mosquito
‘breeding’ habitats to determine its larvicidal effect on immatures of different mosquito
species. Sharma et al. (1998) reported larvicidal activity of Gliricidia sepium against
mosquito larval of A. stephensi, A. aegypti and C. quinquefasciatus. Crude ethanol extracts
of dried leaves, fresh leaves, dried petioles and stem bark of G. sepium against third instar
larvae of A. stephensi, Ae. aegypti and C. quinquefasciatus. Okorie and Lawal (1998)
tested the larvicidal properties of ethanolic extracts of fruits of P. guineense (African black
pepper) on larvae of Aedes aegypti (L) at different concentrations.
The extracts of Cryptotaemia Canadensis, both chloroform and water were
bioassayed against fourth instar larvae of C. pipiens at concentrations between 5 and
50 ppm (Eckenbach et al., 1999). The larvicidal activity of petroleum ether extract of root
of Glycosmis pentaphylla on the juveniles of C. quinquefasciatus, A. stephensi and
A. aegypti was determined (Latha and Ammini, 1999). Al-Dakhil and Morsy (1999) tested
the larvicidal actions of three ethanol extracts of peel oils of lemon, grape fruit and naval
orange against the early fourth instar larvae of C. pipiens and the resulting pupal stages.
Pushpalatha and Muthukrishnan (1999) studied the efficacy of two tropical plant
extracts for the control of mosquitoes. The larvicidal activity of seed and leaf extracts of
Calophyllum inophyllum and leaf extracts of Rhinacanthus nasutus on the juveniles of
Culex quinquefasciatus, Anopheles stephensi and Aedes aegypti was determined. Ethyl
acetate soluble fractions of C. inophyllum and a petroleum ether fraction of R. nasutus
extracts showed very high larvicidal activity.
Thomas and Callaghan (1999) reported the use of garlic and lemon peel extracts as
Culex pipiens larvicides. Both garlic and lemon were toxic to mosquitoes. Latha et al.
(1999) evaluated petroleum ether extracts of 41 indigenous plants abundant in different
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 10
parts of Kerala, India, were studied for larvicidal activity against Culex quinquefasciatus
and C. sitiens. The extracts of 18 plants showed larvicidal activity against the fourth instar
larvae of C. sitiens. Thyagaraj (1999) reported the effect of neem based insecticides against
the mosquito (Culex quinquefasciatus) larvae.
Sagar et al. (1999) reported the bioactivity of ethanol extract of Karanja seed coat
against mosquitoes. Treatment of A. aegypti and C. quinquefasciatus larvae with ethanol
extract of P. glabra seed coat significantly increased the larval mortality and development
period in proportion to increases in the extract concentrations Aedes proved more sensitive
to the effect of extract in terms of mortality than Culex. Markouk et al. (2000) studied the
larvicidal properties of 16 extracts of four Moroccan medicinal plants, C. procera, Cotula
cinerea, S. sodomaeum and S. elaeagnifolium were tested against A. labranchiae mosquito
larvae.
Dwivedi et al. (2000) examined the acetone extracts of leaves of four plants,
Jasminum arbores, Eucalyptus rudis, Bignomia carpreolata and Acacia nilotica were
against A. aegypti. The extracts proved to be potent larvicides causing complete
developmental arrest of fourth instar larvae. Obutour and Onajobi (2000) evaluated the
aqueous and n-butanol fractions of methanolic extracts of Raphia hookori fruit mesocarp
for cytotoxic properties. The n-butanol fraction exhibited greater lytotoxity than the
aqueous fraction when tested against C. quinquefasciatus.
Ignacimuthu (2000) studied the larvicidal activity of 50 medicinal plants collected
from Western Ghats. Results revealed the Streblus aspera, Vitex negunda, Phyllanthus
deblis, L. aspera, Evolvulus aljinoides and Chloroxylon swietenia showed strong larvicidal
activity against the filarial vector C. quinquefasciatus. Crude acetone extract of seeds of
A. squamosa (L) against third and fourth instar larvae of laboratory strain of
C. quinquefasciatus indicated a dose dependent larvicidal and pupicidal activity.
Venkatachalam and Jebanesan (2000) tested the plant parts of 156 plants, extracted
with different solvents. The results indicated that the plant extracts acted as general
toxicants and specifically as larvicides, insect growth regulators, repellents and adulticides.
Rahuman and Venkatesan (2000) assessed the larvicidal activity of ethyl acetate, acetone,
methanol and petroleum ether extract of five indigenous plants, namely Acacia arabica,
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 11
D. alba, Morinda citrifolia, Mukia scabrella and Zingiber officinale against the fourth
instar larvae of C. quinquefasciatus.
Nicolescu et al. (2000) studied the effects of 82 species of plant extracts against
larvae and adults of mosquitoes, A. aegypti and A. atropareus. These plant extracts acted as
toxicants, growth, developmental and reproduction inhibitors and repellents. Lee (2000)
found methanol fruit extract of Piper longum active against larvae of C. pipiens pallens at
10 mg/ml after 24 h. A piperidine alkaloid, piperenon alanine was responsible for this
activity with 24 h median lethal dose (LD50) value of 21 mg/litre.
Ansari et al. (2000) tested the oil of M. piperita for larvicidal activity against
A. aegypti, A. stephensi and C. quinquefasciatus. Of the three species tested
C. quinquefasciatus was most susceptible followed by A. aegypti and A. stephensi.
Rahuman et al. (2000) examined the bioassay guided fractionation of the acetone extract of
Feronia limonia dried leaves afforded a potent mosquito larvicide, which was identified as
n-hexadecanoic acid and found to be effective against fourth instar larvae of
C. quinquefasciatus, A. stephensi and A. aegypti. Namrata et al. (2000) studied the
larvicidal effects of essential oil extracted from the leaves of four plants namely Tagetes
erecta, Ociumum sanctum, Mentha piperita and Murraya koenigii were evalued against
Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti.
Ciccia et al. (2000) studied the insecticidal activity of 11 extracts from nine South
American medicinal plants using the A. aegypti larvicidal assay. Eight of the 11 plant
extracts studied showed toxicity against the A. aegypti larvae. Mehra and Hiradhar (2000)
studied the crude acetone extract of seeds of Annona squamosa Linnaeus tested against
third and fourth instar larvae and pupae of a laboratory strain of C. quinquefasciatus.
Latha and Ammini (2000) reported the leaves and tubes of Curcuma raktakanda as
the mosquito larvicide against the early fourth instar larvae of C. quinquefasciatus,
C. sitiens, A. aegypti and A. stephensi. The petroleum ether extract of the leaves and tuber
exhibited toxicity towards all the test species. Huang et al. (2000) examined a preliminary
study on the toxicity of insecticidal plant against the mosquito larvae of Culex pipiens
pallens.
Momin and Nair (2001) analysed the methanolic extract of Apium graveolens seeds
for bioactive compounds and isolated the mosquitocidal, nematicidal and antifungal
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 12
compounds, sedanolide (1), senkyun-olide-N (2) and senkyunolide – J (3). Crude aqueous
extracts of dried fruit pericarp, flower, root and stem of solenostemma argel were tested for
larvicidal activity against the third instar larvae of the mosquito, C. quinquefasciatus.
Extracts of the fruit pericarp was most effective (El-kamali, 2001). The larvicidal activity
of the plant, Hydrocotyle javanica against C. quinquefasciatus was evaluated using
petroleum ether, benzone, chloroform, ethylacetato and methanol (Venkatachalam and
Jebanesan, 2001).
Redwane et al. (2002) studied the efficacy of extracts and fractions of
Quecuslusitania infectoria galls against second and fourth instar larvae of C. pipiens. The
methanolic, chloroform and ether extracts of Euphorbia tirucalli latex and stem bark were
evaluated for larvicidal activity against C. quinquefasciatus.
Vahitha et al. (2002) reported larvicidal efficacy of leaf extracts of Pavonia
zeylanica and Acacia ferruginea was tested against the late third instar larvae of
C. quinquefasciatus. Jaybala and Rajkumar (2003) determined the larvicidal activity of
plant extracts of D. metal (seed), Cinnamomum zeylanicum (leaf), Baliospermum
montanum (seed) in A. stephensi.
Prabhakar and Jebanesan (2004) tested the larvicidal efficacies of extracts of five
species of Cuccubitacious plants, Momordica charantia, Trichosanthes anguina, Luffa
acutangula, Benincasa cerifera and Citrullus vulgaris against the late third larval stage
group of C. quinquefasciatus. Rajkumar and Jebanesan (2004) evaluated the toxicity of the
plant Moschosma polystachum on C. quinquefasciatus. The crude leaf extract and active
compound octacosane showed negligible mortality against early third instar larvae of
C. quinquefasciatus. Cheng et al. (2004) assessed the chemical position of leaf essential
oils from eight provenances of indigenous Cinnamon (Cinnamomum osmophloeum Kanch)
were compared. The larvicidal activities of leaf essential oils and their consitutents from
the five chemotypes of indigenous cinnamon trees were evalued by mosquito larvicidal
assay.
Chapagain and Wiesman (2005) reported the larvicidal activity of the fruit
mesocarp extract of B. aegyptiaca and its saponin fractions against A. aegypti. Fruit
mesocarp extract of B. aegyptiaca was tested against the laboratory-reared third instars
larvae and compared with their LC50 values.
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 13
Govindarajan et al. (2005) examined the larvicidal effect of extra cellular
secondary metabolites of different fungi against the mosquito, C. quinquefasciatus. The
culture filtrates of five different fungi (A. flavus, A. parasiticus, Penicillium falicum,
Fusarium vasinfectum and Trichoderma viride were tested for the larvicidal activity
against third instar larvae of mosquito vector.
Nath et al. (2006) studied the methanol extracts of 19 indigenous plants as
mosquito larvicide. Among these, pericarp of Zanthoxylum limonella was found to have
the most promising larvicidal properties against A. albopictus and C. quinquefasciatus.
Das et al. (2007) studied the larvicidal activity of methanol and ethanol extracts of
five aromatic plant species against Ae. Albopictus and C. quinquefasciatus larvae varied
according to plant species. Methanol extract of Aristolochia saccata roots was found to be
the most effective against Ae. albopictus larvae.
Mustafa and Al-khazraji (2008) investigated the effects of the extracts of eight
plant species collected from Ninavah governorate on the second instar stage of Culex
pipiens molestus Forskal. Three out of the eight plant extracts Azadirachta excelsa Jack.
Cleome glaucescens Dc. and Quercus infectoria caused 100% mortality of larvae at a
concentration of 200 g/ml after 3 days of treatment. Mustafa (2005) reported that the
seeds extract of Pimpinella anisum L. has high toxicity to second instar larvae of C. pipiens
molestus.
Maheswaran et al. (2008) studied that larvicidal activity of Leucas aspera against
the larvae of Culex quinquefasciatus and Aedes aegypti. The hexane extract of Leucas
aspera showed highest larvicidal activity against the two vector mosquitoes followed by
chloroform and ethanol.
Cheng et al. (2008) examined the mosquito larvicidal activities of methanolic
extracts from different plant parts of red heartwood-type Cryptomeria japonica against the
fourth-instar larvae of Aedes aegypti and Aedes albopictus. Results showed that the larval
mortality of the methanolic extracts of C. japonica heartwood, back and leaf extracts did
not exceed 50.0% indicating no significant toxicity to A. aegypti and A. albopictus, while
the sapwood caused 100% larval mortality in 24 h.
Sathishkumar and Maneemegalai (2008) investigated the larvicidal effect of
Lantana camara extract against early third and fourth instar larvae of Aedes aegypti and
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 14
Culex quinquefasciatus. Extracts from leaves, flowers and roots of plants and oils were
found to have mosquito larvicidal activity.
Senthil Kumar et al. (2009) used the powders of neem seed, leaf, gum, flower, bark
and root for studying antipupational, adulticidal and adult repellency properties against
A. stephensi. Rahuman et al. (2000) undertook bioassay studies using the acetone extract of
Feronia limmia leaves. This extract served as a potent mosquito larvicide and was effective
against fourth-instar larvae of C. quinquefasciatus, A. stephensi and A. aegypti. Row et al.
(2009) examined the essential oil and methanolic and aqueous extracts of Piper betle L. for
their antimicrobial activity, mosquito larvicidal activity, antioxidant property and
mushroom grosinase inhibition. The methanolic and aqueous extracts showed strong
activity against the yeasts: C. albicans and M. Pachydermatis.
Deore and Khadabadi (2009) revealed that the laboratory studies carried out to
ascertain that larvicidal properties of Chlorophytum borivilianum sant. and Fernand
belonging to family. Liliceae is a very well known plant for its aphrodisiac as well as
immunomodulatory properties (Oudhia, 2001). Roots of the plant are used both in
Ayurveda and Unani system to treat oligospermia, arthritis, diabetes and dysuria (Wealth
of India, 1996). Anticancer (Arif, 2005), immunomodulatory (Singh et al., 2004), anti-
diabetic (Govindarajan et al., 2005), antistess (Gopalkrishna and Patil, 2006), aphrodisiac
(Thakur et al., 2006), antimicrobial (Deore and Khadabadi, 2007) and anti inflammatory
(Deore et al., 2008) activities of root extracts have been evaluated. Roots of this plant
contain carbohydrates, phenolic compounds, saponins and alkaloids (Deore et al., 2008).
Samidurai et al. (2009) studied larvicidal, ovicidal and repellent activities of
Pemphis acidula Forst (Lythraceae) against Filarial and Dengue vector mosquitoes. The
larval mortality was observed after 24 h exposure.
Dua et al. (2009) observed that larvicidal activity of neem oil formulation against
mosquitoes. Kihampa et al. (2009) examined the larvicidal activity of seventeen Tanzanian
plant species against the malaria vector. Anopheles gambiae S.S. Giles larvae. The crude
extracts from the leaves, stems and root barks of the investigated plants were obtained by
solvent extraction and then bio-assayed following WHO protocols.
Aina et al. (2009) studied the efficacy of some plant extracts on An. gambiae
mosquito larvae. The ethanolic and aqueous extracts of the fruits of Physalis angulata,
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 15
Xylopia aethiopica and seed of Piper guineense schum and Jatropha curcas Linn. were
tested on the second instar larvae of A. gambiae (L) at varying concentrations.
Borah et al. (2010) studied the larvicidal efficacy of nature fruits and leaves of
Toddalia asiatica against the larvae of A. aegypti and C. quinquefasciatus.The hexane,
acetone and methanol extracts of nature fruits and leaves were investigated to establish its
bio-control potentiality under laboratory condition against fourth instars larvae of
A. aegypti and C. quinquefasciatus.
Patil et al. (2010) studied the larvicidal activity of crude chloroform,
dichloromethane and methanol extracts of the leaves and roots of the six Indian plants,
Aegle marmelos L., Balanites aegyptica L., Calotropis gigantica L., Murraya koenigii L.,
Nyctanthes arbor-tristis L. and Plumbago zeylanica L. were tested against the early fourth
instar larvae of Aedes aegypti L. and Anopheles stephensi. The larval mortality was
observed after 24 h of exposure. All extracts showed moderate larvicidal effects.
Tomass et al. (2011) studied that larvicidal effects of Jatropha curcas L. against
Anopheles arabiensis. The LC50 and LC90 values of Jatrophas curcus crude methanol leaf
extract were found respectively.
Mehdi et al. (2011) assessed the larvicidal and IGR properties of leaf extract of
Cassia fistula and Saraca indica. The early fourth instar larvae of mosquito were exposed
to acetone leaf extracts of Saraca indica and Cassia fistula. Leaf extracts showed moderate
to high larvicidal activity. Zewdneh et al. (2011) observed that larvicidal effects Jatropha
curcas L. against Anopheles arabiensis.
Adediwura et al. (2011) investigated the larvicidal activities of the petroleum ether,
chloroform fractions and methanol extract of Buchholzia coriacea seed as potential agent
in vector control for malaria. The extract and fractions of B. coriacea tested showed that
the chloroform fraction was more effective against the third and fourth instar larvae of
A. gambiae than the petroleum ether fraction and methanol extract. Manilal et al. (2011)
assessed the laboratory and field evaluation of some chemical and biological larvicides
against Culex spp. Tomaphos Bti and Phyriproxyfen.
Maragathavalli et al. (2012) evaluated the mosquito larvicidal activity of leaf
extract of neem. Larvicidal effect on third and fourth instar larvae of Aedes aegypti and
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 16
Culex quinquefasciatus were tested. Rabha et al. (2012) reported the aqueous solutions
obtained during the stream distillation of medicinal and aromatic plants. The hydrolates of
four plants Zanthoxylum limonella, Zingiber officinale, Curcuma longa and Cymbopogon
citrates were evaluated for their larvicidal activity against two laboratory reared mosquito
species Aedes albopictus and Culex quinquefasciatus.
Poonguzhali and Nisha (2012) studied the larvicidal activity of two seaweeds, Ulva
fasciata and Grateloupia lithophila against mosquito vector Culex quinquefasciatus.
Increasing documentation of negative environmental and health impact of synthetic
insecticides and increasingly stringent environment regulation of pesticides have resulted
in the development and use of bioinsect management products for controlling mosquitoes
and pest. Of the two algae screened G. lithophila was found to effective against the larva
Culex in all the three extracts methanol, acetone and benzene.
Anstrom et al. (2012) evaluated mosquitocidal properties of natural product
compounds isolated from Chinese herbs and synthetic analogs of curcumin and their ability
to inhibit binding of cholesterol of Ae. Aegypti sterol carrier protein – 2 in vitro. Kumar
et al. (2012) determined the effectiveness of seaweed (Sargassum wightii) extract
combined with Bacillus thuringiensis val. Israclensis for the control of Anopheles
sundaicus Liston, a malaria vector that occurs in the coastal areas of Peninsular India.
Nour et al. (2012) reported larvicidal activity of extracts from different parts of
neem (Azadirachta indica) against Aedes aegypti mosquitoes. The toxic effect of different
solvents (acetone, chloroform, ethanol) extracts from different parts (bark, leaf, root and
seed) of neem, Azadirachta indica against A. aegypti larvae.
Illahi et al. (2012) examined larvicidal activities of different parts of Melia
azedarach Linn. against Culex quinquefasciatus. Various concentrations (50, 100, 500,
1000, 1500 and 2000 ppm) of aqueous extracts of leaves, fruits and bark of Melia
azedarach were tested for larvicidal activity against Culex quinquefasciatus. There
occurred a continuous increase in mortality of third and fourth instar larvae with increase
in concentration of the extracts.
Sakthivadivel et al. (2012) studied the evaluation of toxicity of plant extracts
against vector of lymphatic filariasis, C. quinquefasciatus. The toxic effects of petroleum
ether leaf extracts of plants viz., Argemone mexicana, Clausena dentala, Cipadessa
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 17
baccifera, Dodonaea angustifolia and Melia dubia were evaluated under laboratory
conditions individual an in combination against third instar and fourth instar larvae of
Cx. Quinquefasciatus.
Illahi and Ullah (2013) reported the larvicidal activity of different parts of
Artemisia vulgaris Linn. Against C.quinquefasciatus. The third and fourth instar larvae of
C.quinquefasciatus were exposed for 24 hrs to various concentrations (50, 100, 500, 1000
and 1500 ppm) of methanol extracts of different parts of Artemisia vulgaris.
2.2. Antimicrobial ActivityNaseimento et al. (2000) investigated that antimicrobial activity of plant extracts
and phytochemicals was evaluated with antibiotic susceptible and resistant
microorganisms. Extracts like Achillea millifolium, Caryophyllus aromaticus, Melissa
officinalis, Ocimun basilucum, Psidium ghajava, Punica granatum Rosmarinus officinalis,
Salvia officinalis, Syzygyum joabolanum and Thymus vulgaris were analysed.
Adeleye et al. (2003) tested antimicrobial activity of some local herbs on common
skin pathogens. Extracts from Anogeissus leoicarpus, Daniellia diveri and Xylopia
aethiopica were screened for antimicrobial activity against Staphylococcus aureus,
Candida albicans, P. aeruginosa, Trichophyton mentagraphytes and As. fumigatus using
disc and well diffusion methods. Both aqueous and ethanolic extracts of the three plants
showed antimicrobial activities against two microorganisms.
Lai and Roy (2004) reported the antimicrobial activity of plant extracts may reside
in a variety of different components, including aldehyde and phenolic compounds.
Naturally occuring combinations of these compounds can be synergistic and often result in
extracts having greater antimicrobial activity than the purified individual constituents
(Delaquis et al., 2002).
Shanab et al. (2004) studied that antibacterial activities of hot water, methanol and
ethanol extracts of five plants extracts utilized in Palestine. The dried extracts of Syzyium
aromaticum, Cinnamomum cassia, Salvia officinalis, Thymus vulgaris and Rosmarinus
officinalis were tested in vitro against 4 bacterial species by disk diffusion and micro-
dilution. The patterns of inhibition varied with the plant extract, the solvent used for
extraction, and the organism tested. Methicillin resistant Staphylococcus aureus and
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 18
Bacillus subtilis were the most inhibited microorganisms. S. aromaticum extract was the
most active against multidrug resistant P. aeruginosa and enterohemorrhagic E. coli.
Owoyale et al. (2005) observed that the methanolic, ethanolic and petroleum ether
extracts of Senna alata leaves for phytochemical, antibacterial and antifungal activities.
The phytochemical screenined revealed that most active chromatographic component is a
flavonoid glycoside. Senna alata leaves crude extract showed both antibacterial and
antifungal activities. The methanolic extract was the most active of the three crude extracts
tested against bacterial and fungal organisms. It is appeared that the most active
chromatography component is only slightly effective both in degree of inhibition and
spectrum of activity on all the test organisms to the methanolic crude extract.
Prabuseenivasan et al. (2006) reported the antibacterial activity of 21 plant essential
oils against six bacterial species. The selected essential oils were screened againts four
Gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, Pseudonomonas
aeruginosa, Proteus vulgaris) and two Gram positive bacteria Bacillus subtilis and
Staphylococcus aureus at four different concentrations. Doughari et al. (2006) reported
antimicrobial activity of Tamarindus indica. The antimicrobial activities of extracts of the
stem bark and leaves were evaluated against Gram negative, Gram positive bacteria and
fungi.
Egwaikhide and Gimba (2007) studied the analysis of the phytochemical content
and anti-microbial activity of Plectranthus glandulosis by using hexane, ethyl acetate and
ethanol. The ethyl acetate extract of Plectranthus glandulosis is the most active, showing
activity against three Gram negative and three Gram positive bacterial strains. Ushimaru
et al. (2007) demonstrated antibacterial activity of medicinal plants extracts against E. coli,
Salmonella typhimurium, Staphylococcus aureus and Enterococcus species. The
methanolic extract of Caryophyllus aromaticus exhibited the highest anti S. aureus activity
and was effective against all bacterial strains tested.
Sokovic et al. (2007) examined the chemical composition and antibacterial activity
of essential oils from 10 aromatic plants namely Matricaria chamommilla, Mentha
piperita, M. spicata, Lavandula angustifolia, Origanum vulgare, Salvia officinalis, Citrus
limon and C. aurantium have been determined. Antibacterial activity of these oils and their
components, were assayed against a variety of human pathogenic bacteria. The highest and
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 19
broadest activity was shown by Origanum vulgare oil. Carvacrol possessed the highest
antifungal activity among the components tested.
Parekh and Chanda (2007) evaluated the antibacterial effect of some selected
Indian medicinal plants on bacterial strains like Bacillus cereus, Staphlococcus aureus,
Enterobacter aerogenes, Escherichia coli and Klebsiella pneumoniae. The solvents used
for the extraction of plants were water and methanol. The most susceptible Gram-positive
bacteria was B. cereus, while the most susceptible Gram-negative bacteria was
K. pneumoniae. The extracts of Abrus precatorius, Cardiospermum lialicacabum and
Amelina asiatica could not inhibit any of the bacteria strains investigated. The most active
antibacterial plant was Caesalpinia pulcherrima.
Parekh et al. (2007) examined the antibacterial activity of 12 plant extracts against
six bacterial strains. The aqueous extract of Caesalpinia pulcherrima, Casuarina
equisetifolia, Euphorbia hirta showed antibacterial activity. The methanol extracts of all
the plants exhibited less antibacterial activity. Doughari et al. (2007) studied the bioactive
compounds of root extracts of Carica papaya, using water and organic solvent, and were
investigated for antibacterial activity against some pathogenic bacteria using the cup plate
agar diffusion method.
Mahesh and Sathish (2008) examined that methanol leaf extracts of Acacia nilotica,
Sida cordifolia, Tinospora cordifolia, Withania somnifer and Ziziphus mauritiana had
significant antibacterial activity against Bacillus subtilis, Escherichia coli, Pseudomonas
fluorescens, Staphylococcus ameus and Xanthomonas axonopodis, Paramathivelur.
Malvacearum and antifungal activity against Aspergillus flavus, Dreschlera turcica and
Fusaitum verticillioides when compared to root / bark extracts.
Doughari et al. (2008) stated the antimicrobial activity of leaf extracts of Senna
oblusifolia against both clinical and laboratory isolates of both bacteria and fungi using the
disc diffusion method. Mathad and Mety (2010) evaluated phytochemical and
antimicrobial activity of Digera muricata. Among the bacteria used viz. (K. pneumonia,
E. coli, S. aureus, S. typhi and V. cholerae) showed the petroleum ether extract showed
highest zone of inhibition and similarly in fungi (As. flavus, A. niger, A. terreus and
C. albicans) ethanol extract exhibited highest zone of inhibition.
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 20
Philip et al. (2009) examined antimicrobial activity of some medicinal plants
namely Pereskia bleo, Pereskia grandifolia, Curcuma aeruginosa Roxb, Curcuma zedoria,
Curcuma mangga, Curcuma Tnodora off. Blatter, Zingiber officinale vcu, Officinale and
Zingiber officinale val. rubrum were screened for their antimicrobial activity against both
Gram positive bacteria and Gram negative bacteria using agar disc diffusion assay.
Krishnaiah et al. (2009) studied the antibiotic resistance that develop from
prolonged usage of certain drugs has led to continuous efforts in searching for metabolites
that possess antimicrobial activities. Plants secondary metabolites (alkaloids, terpenoids
and phenolic compounds) are potential antimicrobial agents that can help to alleviate
problem of antibiotic resistance.
Row and Ho (2009) studied the antimicrobial activity, mosquito larvicidal activity,
antioxidant property and Tyrosinase inhibition of piper betle. The methanolic and aqueous
extracts showed strong activity against the yeasts: C. albicans and M. pachydermatis. The
crude essential oil exhibited a broad spectrum strong antimicrobial activity against all test
organisms. The strong activity was observed against C. albicans, followed by S. aureus
and M. pachydermatis.
Lelono et al. (2009) examined isolation antifungal compounds from Gardenia
jasminoides. Methanol extracts showed the highest level of antifungal activity against
Pleurotus ostreatus, a wood-rotting fungus, compared to five other methanol plant extracts
(Thuja orientalis L, Datura innoxia, Ligustrum jasponicum T, Juniperus chinensis var.
Procumbens and Mallotus japonica M.
Sivanarayan and Suriyavathana (2010) examined preliminary studies on
phytochemicals and antimicrobial activity of Delonix elata and Prosopis cineraria. The
invitro antimicrobial activity was performed by agar disc diffusion method against
bacterial viz. Staphylococcus aureus, Bacillus subtilus, Klebsiella, E. coli, Proteus sp. and
Sp. and fungi viz. As. niger, Pencillium sp. Candida albicans. Arun et al. (2010) assessed
the invitro antibacterial activity and flavonoid contents of Lawsonia innermis.
Khan and Nasreen (2010) reported the phytochemical analysis, antifungal activity
and mode of action of methanol extracts from plants against pathogens. Among all
extracts, L. innermis showed greatest inhibition of mycelial growth of target fungi followed
by Withania somnifera.
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 21
Mostafa et al. (2011) examined antifungal and antiaflatoxigenic activities of five
methanolic plant extracts against the anti-aflatoxigenic activities followed by Zingiber
officinalis. P. granatum extract absolutely arrested aflatoxin B1 production at 5 mg/ml and
inhibited 100% the mycelial growth of A. flavus at 10 mg/ml. Zingiber officinalis and Olea
europaea extracts showed a moderate antifungal activities and exhibited a significant anti
aflatoxigenic efficiency as they completely inhibited aflatoxin B1 production at 15 mg/ml.
Lantana camara extract showed a weak antifungal activities while no effect was detected
with Allium sativum L.
Dash et al. (2011) studied the medicinal and antibacterial activities of methanol and
acetone extracts of the two species Trigonella foenum L. and Coriandrum sativum L.
available in Bangladesh. Crude extract of the spices with methanol and acetone were
screened for antibacterial activities against four Gram negative pathogenic bacteria.
Pseudomonas spp., E. coli, Shigella dysentiriae and Salmonella typhi. The invitro
antibacterial activity was performed by T. foenum and sativum revealed an elevated
antimicrobial activity against Pseudomonas spp. whereas acetone extract of spices
exhibited highest activity against E. coli.
Rayavarapu and Kaladhar (2011) evaluated the antibacterial activity of hexane,
chloroform and methanol extracts of plant extract from Mucuna pruriens leaves against
Erwinia carotovora, Pseudomonas syringae, Pseudomonas marginalis, Pseudomonas
aeruginosa, Xanthomonas campestris. Study of antifungal activity of whole plant extract of
Mucuna pruriens against Curvularia lunata, Fusariumoxy sporum, Pencilium expansum.
Rhizoctonia solani, Tiarosporella phaseolina and Ustila gomaydis. All extracts exhibited
antimicrobial activity against different species of bacteria and fungi. The methanol extract
of Mueuma pruriens showed highest antimicrobial activity against all the bacterial and
fungal species tested, whereas chloroform extracts showed moderate antimicrobial activity
against both bacterial and fungal species.
Elumalai et al. (2011) assessed the antibacterial activity of various leaf extracts of
Merremia emarginata. The antibacterial activity and phytochemical screening of the
aqueous, methanol and petroleum ether leaf extracts were evaluated by agar well diffusion
method against four bacterial species. The methanol extract was more effective against
Bascillus cereus and E. coli, whereas aqueous extract was more effective against
Staphylococcus aureus and P. aeruginosa.
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 22
Zerroug et al. (1998) studied the antibacterial activity of extracts of Ajuga iva and
Teucrium polium. The methanolic extracts of these plants were tested against five bacterial
species E. coli, B. subtilis, P. diminutus, Paracoccus paratrophus and Micrococcus luteus.
Kim et al. (2011) evaluated antimicrobial activity of 12 plant extracts against
Salmonella typhimurium, E. coli and Listeria monocytogenes by using a disc diffusion
assay, and Syzygium aromaticum (clove) showed the highest inhibitory effect. The efficacy
of clove extract inactivates pathogens on lettuce, inoculated lettuce with S. typhimurium,
E. coli and L. monocytogenes were treated with diluted clove extracts or distilled water for
0, 1, 3, 5 and 10 min. clove extract treatment significantly reduced populations of the
3 tested pathogens from the surface of lettuce.
Singh (2011) illustrated antimicrobial effect of callus and natural plant extracts of
Premna serratifolia. It is an important medicinal shrub used in traditional system of
medicines as cardio toxic, antibiotic, anti-coagulant, carminative, hepato protective,
antitumor etc. Evalution of antimicrobial activity against the selected human pathogens
Bacillus sp., Enterococcus faecalis, E. coli, Klebsilla pneumonia, Non-haemolytic
streptococci, Streptococcus epidermidis, P. aeruginosa and Salmonella typhimurium by
using natural leaves, roots and its respective calli induced with help of various plant growth
regulators.
Madhumathi and Rajendran (2011) examined the antimicrobial activity of leaf
extract of Passiflora incarnate. The ethanolic extract of leaves was screened for
phytochemical constituents and antimicrobial activities towards bacteria and fungi. The
phytochemical analysis of the P. incarnate leaf extract shows the presence of tannins,
alkaloids, flavonoids and phenolic compounds. Tannins have been found to form
irreversible complexes with proline-rich proteins resulting in the inhibition of the cell
protein synthesis. The leaf of P. incarnate were effective against bacteria and fungi.
Adu et al. (2011) studied the effects of Acanthosperum hispidum extract on the
antibacterial activity of amoxillin and ciprofloxacin. The effect was assayed by disc
diffusion method. The extract demonstrated antibacterial activity against S. aureus,
Klebsiella pneumoniae, Bascillus subtilis, Salmonella thypi, Proteus vulgaris and
P. aeruginosa with MICs ranging between 11 and 53 mg/ml. 5 mg/ml extract enhance
significantly (p < 0.05) ciprofloxacin activity (upto 38 folds) against all the test bacteria
except P. aeruginosa.
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 23
Wendakoon et al. (2011) examined the evaluation of selected medicinal plants
extracted in different ethanol concentration for antibacterial activity against human
pathogens. Penmus boldus, Agathosma betulina, Echinacea angustifolia, Humulus lupulus,
Glycyrrhiza glabra, Mahonia aquifolium, Usnea barbata and Anemophsis california were
screened for antibacterial activity against four Gram-positive and Gram-negative
pathogens.
Sumathi et al. (2011) observed the antibacterial activity of the plant extract of S.
officinale against selected pathogenic bacteria. The effect of ethanol, methanol, chloroform
and ethyl acetate extract of S. officinale leaves were investigated by streak plate method for
antibacterial activities against six bacterial strains. The result showed that the ethanol, ethyl
acetate and methanol extracts of leaves of S. officinale possess measurable antibacterial
activities against all the bacterial strains, while chloroform extracts of the plant did not
produce any measurable antibacterial activity.
Misra et al. (2011) studied the antibacterial activity of roots, stembark and leaves of
various solvent (Hexane, Benzene, Isopropanol, Ethylacetate, Methanol and Water)
extracts of Alstonia scholaris using well diffusion techniques on Gram positive
(Staphylococcus aureus, Bacillus cereus and Lactococcus lactis) as well as Gram negative
bacteria (Aeromonas species, Enterobacter aerogenes, Escherichia coli, Pseudomonas
aeruginosa and Proteus mirabilis).
Soyelu and Masika (2012) reported antibacterial and antioxidant activities of some
selected plants used for the treatment of cattle wounds. 1-Diphenl-2-picrycl-hydrazyl
(DPPH) radical scavenging activity of the methanol and water extracts of A. ferox,
P. obliquum and C. aurea when compared with the standard (BHT) butylated
hydroxytoluene at varying concentrations.
Parameswari et al. (2012) studied the invitro antibacterial activity in the extracts of
Solanum nigrum. The anti-bacterial activity of ethanolic and methanolic extracts of stem,
berries and whole plant of S. nigrum was detected against Bacilus subtilis, E. colia,
Klebsiella pneumonia and P. aeruginosa. S. nigrum plant extracts of 10 g, 50 g and
100 g were taken as per CLSI standard control streptomycin. The methanolic extracts
were showed highest antibacterial activity than ethanolic extracts.
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 24
Arora et al. (2012) studied the total phenolic content and antimicrobial activity of
seed, pomace and leaf extracts of sea buckthorn (Hippophae rhamnoidas L.) against food
borne and food spoilage pathogens. The methanolic extract of leaves exhibited high total
phenolic content (278.80 mg GAE/g extract) and minimum inhibitory concentration (MIC)
value of 125 g/ml against Listeria monocytogenes.
Subedi et al. (2012) assessed the methanol extract of the bark of Machilus
odoratissima and its antioxidant and antibacterial properties. The phytochemical screening
demonstrated the presence of different types of compound like terpenoids, tannins, deoxy
sugar, saponins and phenolic compounds.
Gautam et al. (2012) evaluated the antibacterial potential of petroleum ether,
acetone, methanol and aqueous extracts of Nepeta ciliaris against selected respiratory tract
pathogens. The extracts from the aerial parts of N. ciliaris at concentration of 200mg/ml
were screened against three Gram-positive (Staphylococcus aureus MTCC 1144,
Streptococcus pneumonia MTCC 655 and Streptococcus pyogens MTCC 442) and one
Gram-negative (Pseudomonas aeroginosa MTCC 2474) bacterial pathogens. The agar well
diffusion method was adopted to examine antibacterial and minimum inhibitory
concentration (MIC) values of most effective extracts against the susceptible bacteria.
Erythromycin was used as positive control to determine the sensitivity of the strains.
Mahmood and Qureshi (2012) investigated the antimicrobial activities of crude
mathanolic extract of leaves of Acacia nilotica L., Albizia lebbeck L. and activity was
studied by agar well diffusion method against one Gram-positive B. subtilis and three
Gram-negative P. aeruginosa, E. coli and Klebsiella pneumonia. Crude extract of all plants
showed best activity against Gram-negative bacterial strains while minor inhibition zones
were found against Gram-positive bacterial strains. Antifungal activity of crude plant
extract was screened by agar tube dilution method against Aspergillus nigar and
Aspergillus flavus.
Zain et al. (2012) studied the animicrobial activities of Saudi Arabian desert plants.
The ethanol extracts of Alhag maurorum Medic., Cheropodium murale L., Convolvulus
fatmensis G. Kunze, Conyza dioscoridis (L). Desf., Cynandrum acutum L., Diplotaxis acris
(Forssk) Boiss, Euphorbia cuneata Vahl., Origanum syriacum L., Solenostemma argel
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 25
(Del) Hayne and Tamarix aphylla L. (Karst) showed significant antimicrobial activity
against Gram negative, Gram positive bacteria, unicellular and filamentous fungi.
Kasim et al. (2012) assessed the antimicrobial activity of six selected plants against
some strains of pathogenic organisms. Crude ethanolic extracts of selected six Nigerian
plants based on ethnobotanical reports of their antifungal and antibacterial actions were
subjected to phytochemical and antimicrobial screening. All the six plants namely
Acalypha fimbriata, Glaphae brevis, Vernonia amygdalina, Struchium sparganophora,
Celosia argentea and Amaranthus spinosus were screened against strains of Candida
albicans, Trichophyton metagophyte, Malassezia furfur, Aspergillus flavus, Escherichia
coli, Pseudomonas aeruginosa, Salmonella typi and Staphylococcus aureus.
Garba and Okeniyi (2012) examined the antimicrobial activity of the total alkaloids
from five medicinal plants namely Jatropha curcas, Calotropis procera, Magifera indica,
Carica papaya and Psidium guajava used in the treatment of various oilments was
evaluated using disc diffusion assay. The extracts were used against three bacterial isolates
and two fungal isolates including Staphylococcus aureus, Stretococci, Lactobacillus spp.,
Actinomycetes and Candida albicans respectively. The results shows that all the plants
contains various level of alkaloids and an appreciable level of antimicrobial activities were
observed at a concentration level of 6 x 102 g/cm3 indicating that the plants could be a
potential source of alkaloids that may be used for the treatment of various microbial
diseases caused by the tested organisms.
Panthi and Chaudhary (2012) studied the antibacterial activity of some selected
folklore medicinal plants from west Nepal. Eighteen plant species were tested for their
antibacterial activity by the disc diffusion method. The bacteria employed were Gram-
positive (Staphylococcus aureus) and Gram-negative (Escherichia coli, Pseudomonas
aeruginosa and Shigella boydii). Extracts of eight plants showed encouraging results
against three strains of bacteria, while others showed activity against one or two strains.
Saad et al. (2012) investigated the antimicrobial property of mangrove plant
Sonneratia alba (S. alba). The antimicrobial activity was evaluated using disc diffusion
and microdilution methods against six micro organisms. The methanol extract appeared to
be the most effective extract while n-hexane extract showed no activity. The antimicrobial
activities were observed against the Gram-positive bacteria Staphylococcus aureus
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 26
(S. aureus) and Bacillus cereus (B. cereus), the Gram negative E. coli (E. coli), and the
yeast Cryptococcus neoformans, P. aeruginosa and Candida albicans.
Sharmeen et al. (2012) investigated the antibacterial activity of 35 aqueous extracts
against a total of 20 clinical Klebshiella species isolates. The maximum antibacterial
activity was found as 90% in crude extract of Syzygium aromaticum (leaf) and Citrus limon
L. (fruit) followed by 85% in Spondias pinnata (Leaf). Sensitivity of these isolates was
also evaluated for eight commercial antibiotic discs following disc diffusion assay where
most of the isolates found to develop resistance against multiple commercial antibiotics,
85% of isolates exhibited resistant to chloramphenicol and erythromycin and 80% were
resistant to sulfamethoxazole and cephradine. The isolates showed their resistance between
55 – 60% to the other four antibiotic discs, Viz. gentamycin, streptomycin, ciprofloxain
and azithromycin. Among 35 herbal extracts tested, 19 herbal extracts were found to
possess antimicrobial activity in all multi-drug resistant isolates. Therefore these herbal
extracts could be used as alternative therapeutic agents for the treatment of human diseases
caused by Klebsiella species.
Prabhakar et al. (2012) studied the invitro studies on antimicrobial screening of leaf
extracts of Sapindus saponaria against common dental pathogens of Staphylococcus
aureus, Proteus vulgaris, P. aerogenos and Micrococcus abbus. The results of
antimicrobial assay revealed that the extract showed good inhibitory activity against all the
tested pathogens compared with standard antibiotic drug streptomycin.
Subedi et al. (2012) reported the methanol extracts of the bark of Machilus
odoratissima was subjected to investigate its antioxidant and antibacterial properties. The
phytochemical screening demonstrated the presence of different types of compound like
terpenoids, tannins, deoxy sugar, saponins and phenolic compounds. The methanol extract
of the plant was tested for antioxidant activity using scavenging activity of DPPH (1, 1-
diphenyl-2-picrylhydrazil) radical method and antibacterial activity against Staphylococcus
aureus, Enterococcus faecalis, Escherichia coli and Pseudomonas aeruginosa bacteria
using cup plate method.
Mamidala and Gujjeti (2013) studied the phytochemical and antimicrobial activity
of the petroleum ether, chloroform, ethyl acetate and methanol extracts of Acmella
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 27
paniculata and tested for antimicrobial activity against 12 strains of micro organisms using
agar dilution method.
Raji and Raveendran (2013) reported the antifungal activity of five plant extracts
namely Abrus precatorius, Aegle mannelos, Aporosa lindleyana, Areca catechu and
Brassica juncea against phytopathogenic fungi As. niger.
2.3. Phytochemical screening
Oloyede (2005) studied the unripe pulp of Carica papaya to test the presence of
certain phytochemicals. Chemical position of the pulp were determined. Phytochemical
screening of Carica papaya showed the presence of saponins and caldenolides while
chemical analysis revealed the presence of potassium, sodium, calcium, iron, phosphorus,
zinc, copper, magnesium and manganese in considerable quantities.
Egwaikhide and Gimba (2007) examined the extracts of Plectranthus glandulosus
by using hexane, ethyl acetate and ethanol as solvents were screened for secondary
metabolites and antibacterial activity. The extract revealed the presence of alkaloids,
tannins, anthraquinones, glycosides, reducing sugars, saponins, flavonoids, phlobatanins,
steroids and terpenoids.
Chhetri et al. (2008) assessed the phytochemical and antimicrobial evaluations of
some medicinal plants. A qualitative phytochemical analysis was performed for the
detection of alkaloids, glycosides, terpenoids, steroids, flavoids, tannins and reducing
sugar.
Suresh et al. (2008) assessed the antimicrobial and phytochemical investigation of
the leaves of Carica papaya L., Cynodon dactylon (L) Pens, Euphorbia lista L., Melia
azedaluch L., Psidium guajava L. The plant leaves contain a number of medicinally
important compounds. The phytochemical analysis reveals the presence of amino acids, a
amylase, -amylase, carbohydrate, glutamine, protein, proline, phenollic compounds and
the presence of various secondary metabolites such as alkaloids, anthroquinone, catachol,
flavonoids, phenols, saponins, steroids, triterpenoids and tannins were determined.
Asaolu et al. (2010) studied the evaluation of phytochemicals and antioxidants in
the dry leaves of aqueous and methanolic extracts of Vernonina amygdalina, Carica
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 28
papaya, Persea Americana and Cnidosculous aconitifolius. Methanolic extract was found
to contain the highest value for sterols, tannin, saponin, flavonoids, alkaloids, phenols,
phlobatannins, anthraquinones, triterpenes and cardiac glycosides.
Aiyegoro and Okoh (2010) examined the preliminary phytochemical screening and
in vitro antioxidant activities of the aqueous extract of Helichrysum longifoliu. In the
phytochemical analyses revealed the presence of tannins, flavonoids, steroids and saponins.
The presence of gallic acid has been reported in the leaves extract (Manpony et al., 2009)
and in the stem, bark and root extracts (Diwani et al., 2009) of Jatropha curcas plant. The
presence of benzoic acid in the residues of nodes, leaves, stem and root of Egyptian
J. curcas has been reported by Diwani et al. (2009).
Bolla and Bhogavalli (2010) examined the antibacterial properties of Antigonon
leptopus against certain Gram positive bacterial strains (Bacillus subtilis, Bacillus peritolis
and Salmonella typhi) using disc diffusion method. The flower extracts of A. leptopus
were prepared using different solvents like ethanol and chloroform and are screened
for its antibacterial activity.
De et al. (2010) examined phytochemicals present in the tubes extract of
Amorphaphallus paeoniifolius and development of new solvent system for thin layer
chromatography of petroleum ether and ethanol extract of Amorphaphallus paeoniifolius
tuber. The tubes of A. paeoniifolius contain phytoconstituents like alkaloids, steroids, fats
and fixed oil, flavonoids, tannins, proteins and carbohydrates.
Elumalai et al. (2011) examined the antibacterial activity and phytochemical
screening of the aqueous, methanol and petroleum ether leaf extracts of Merrenia
emarginata. The phytochemical screening shows the presence of tannins, flavonoids,
amino acids, starch, glycosides and carbohydrates in the different leaf extracts.
Misra et al. (2011) studied the antibacterial activity of roots, stembark and leaves of
various solvent (Hexane, Benzene, Isopropanol, Ethylacetate, Methanol and Water)
extracts of Alstonia scholaris using well diffusion techniques on Gram positive
(Staphylococcus aureus, Bacillus cereus and Lactococcus lactis) as well as Gram negative
bacteria (Aeromonas species, Enterobacter aerogenes, Escherichia coli, Pseudomonas
aeruginosa and Proteus mirabillis).
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 29
Zohra (2012) reported the triphytochemical screening of three extracts etheric,
ethanolic and aqueous of Malva sylvestris L. The seed contain alkaloids, sterols and
steroids, reducing sugars, tannins, emodols, starch, coumarins and the stem contain
flavonoids, tannins, starch, saponins, alkaloids, emodols, sterols and steroids, reducing
compounds, coumarins and anthocyanosides.
Unnikrishnan et al. (2012) assessed the preliminary phytochemical analysis of bark
of Ficus bengalensis Linn and antibacterial activity of its bark oil. Phytochemical screening
of the bark showed positive tests for sterols, flavanoids, glycosides, saponins and tannins.
Barkatullah et al. (2012) reported the ethanolic acid n-hexane extracts of leaves,
fruits and bark of Zanthoxylum armatum for their antimicrobial potential against bacteria
and fungi. The total flavonoids and phenolic contents were quantified in the ethanolic
extract of all parts. Highest flavonoids was found in ethanolic extract of Z. armatum fruit
followed by ethanolic extract of Z. armatum bark, while highest phenolic contents were
found in Z. armatum fruit followed by Z. armatum bark.
2.4. GC – MS AnalysisIsidorov and Vinogorova (2003) examined GC-MS analysis of compounds
extracted from buds of Populus balsamifera and Populus nigra. In hexane extracts,
54 “neutral” compounds were recorded. The greatest amount of them are sesquiterpenes
and n-alkanes. Among 56 components of ether extracts, many aliphatic acids and
hydroxyacids were detected.
Pichini et al. (2005) examined quantification of the plant-derived hallucinogen
salvinorin A in conventional and non-conventional biological fluids by gas
chromatography / mass spectrometry after Salvia divinorum smoking. The analytes were
extracted from biological matrices with chloroform / isopropanol (a:1, u/v).
Chromatography was performed on a 5% phenyl methyl silicone capillary column and
analytes were determined in the selected ion monitoring mode. The method was validated
over the concentration range 0.015 – 5 g/ml.
Row et al. (2009) examined the essential oil and methanolic and aqueous extracts
of Piper betle L. for their antimicrobial activity, mosquito larvicidal activity, antioxidant
property and mushroom grosinase inhibition. The methanolic and aqueous extracts showed
strong activity against the yeasts, C. albicans and M. Pachydermatis.
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 30
Delazar et al. (2009) studied the GC-MS analysis of Ornithogalum procerum.
A total of 23 compounds were identified from the essential oils of the aerial parts of
O. procerum. Cyclohexane, phenylacetaldehyde, hexa hydrofarnesyl acetone, 5-methyl
octadecane and docosane were the major components of the essential oils and
hexahydrofarnesyl acetone was the most abundant. The n-hexane extract of the bulbs,
seven hydrocarbons representing 99.39% of the total extract were identified and the
hydrolyzed methanolic extract of the bulbs, four polysterol type compounds accounting for
59.81% of the extract were detected.
Claveria et al. (2009) examined metabolic profiling of bioactive Pancratium
canariense extracts by GC-MS AchE inhibitory activities of the alkaloid fractions at a
concentration of 10 g/ml were 29.80 0.91, 40.93 4.60 and 58.06 1.18% for the
bulbs, leaves and fruits respectively. Seventy six metabolites-mono, di and trisaccharides,
fatty acid, amino acids, sterols as well as several Amaryllidaceae alkaloids were detected.
Purification of the alkaloids from the methanolic extracts resulted in the detection of
31 compounds including several potent AchE inhibitors such as habranthine and
galanthamine and the structural elucidation of 3-0 –acetylhabranthine, a new natural
compound with potential AchE inhibitory activity.
Nirmaladevi et al. (2010) stated the analyses of the methanolic extract of leaves of
Rhinacanthus nasutus. The phytochemical constituents of the leaves of Rhinacanthus
nasutus in order to understand the nature of the principle component responsible for its
medicinal property. A preliminary absorbance survey scan of the methanolic extract of
R. nasutus evidenced the presence of multiple components in the extract. Two peaks
observed in the HPLC spectrum showed the presence of two compounds in the extract.
GC-MS profile revealed that the active components present in the leaf extract might be
alkaloids or polyphenols. The results of IR spectrum revealed that band 1 possess
compounds of polyphenolic nature and band 2 possess compounds that are having a
hydroxyl and a carbonyl groups.
Charles et al. (2011) examined the GC MS analysis of bioactive components on the
bark extract of Alseodaphne semecarpifolia. In this GC MS analysis, 28 bioactive
phytochemical compounds were identified in stem bark of Alseodaphne semecarpifolia.
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 31
The 28 components predominantly phenolic derivatives are present included hydrocarbons,
carbohydrates, fatty acid, fatty acidester, alcoholic compounds, alkaloids, ketones and
alkenes compounds. These different active phytochemicals have been found to possess a
wide range of activities, which may help in the protection against incurable diseases.
Ajayi et al. (2011) revealed the gas chromatography mass spectrometry analysis
and phytochemical screening of ethanolic root extract of Plumbago zeylanica. The
phytochemical screening showed the presence of alkaloids, tannins, steroids, flavonoids,
saponins, anthraquinones, cardia-glycosides, phlobatinnins and carbohydrates. The gas
chromatography-mass spectrometry analysis also identified the presence of phytochemical
components like phenolics (Phenol, 2, 4 – bis (1, 1-dimethyl ethyl) (RT: 6.796).
Cyclopentadecane (RT: 7.305), fatty acids / methyl esters, feroprofen (RT: 9.602), 1, 1, 1,
5, 7, 7, 7 – Heptamethyl 3, 3-bis (trimethylsiloxy) tetrasiloxane (RT: 8.253), Indazol-4-one,
3, 6, 6-trimethyl-1-phthalazin-1-y1-1, 5, 6, 7-tetrahydro-(RT: 10.162), 2H-Indol-2-one,
1-(2,6-dichlorophenyl)-1, 3-dihydro-(RT: 11.191), 2-methyl-7-phenylindole (RT: 14.540),
1H-Indole-2-carboxylic acid, 6-(4-ethoxyphenyl)-3-methyl-4- 5, 6, 7-tetrahydro-, isopropyl
ester (11.528), and anthranilic acid, 3-chloro-N-(o-chlorophenyl)- (RT: 12.443).
Eddy et al. (2011) examined the GC MS study of ethanol extract of Andrographis
paniculata revealed the presence of hydrocarbons ((E)-tetradec-3-ene), alcohol (6-methyl
heptan-1-01), aromatics (undecan-5-ylbenzene, dodecan-5yl benzene, undecan-2-
ylbenzene, (2-methyl undecan-6-yl) benzene and dioctyl phthalate). Some of the identified
compounds were found to be green raw materials for polymer industries and
pharmaceutical companies.
Namuli et al. (2011) determined the phytochemical compounds in different solvent
extracts of Jatrapha curcas Linn. Aqueous, methanolic and hexane extracts of various
plant parts were analysed for phytochemical compounds by spectrophotometry, high
performance liquid chromatography (HPLC) and gas chromatography mass spectrography
analysis (GC MS). The root bark methanolic extract contained high phenolics (11.5 / mg
gallic acid equivalents / g DW) and flavonoids (0.94 mg rutin equivalents / g DW) and the
methanolic extract contained 1.13 mg/g DW phorbol esters. Abirami and Rajendran (2012)
examined the bioactive compounds of Vernonia cinerea have been evaluated using
GC-MS. The chemical compositions of the whole plant methanol extract of Vernonia
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 32
cinerea were investigated using Perkin – Elmer gas chromatography mass spectroscopy.
GC-MS analysis of V. cinerea plant methanol extract revealed the existence of the GC-MS
chromatoGram of the seven peaks presented. The major compounds n-hexadecanoic acid
(42.88%) (Retention time 16.26) and 1.2 benzene dicarboxylic acid disoocty ester (23.00)
(Retention time 24.81).
Kucukboyaci et al. (2012) determined gas chromatography of quinolizindine
alkaloids in Genista sandrasica and their antimicrobial activity. Ten quinolizidine
alkaloids were identified in the alkaloid extract of G. sandrasica. The main alkaloids were
spartein (13.68%), N-acetylcytisine (6.48%), -methoxylupanine (13.12%), anagyrine
(40.49%) and baptifoline (10.76%).
The chemical composition of the essential oil and its fractions was analyzed by
GC/MS analysis, Eugenol (36.2%), Chavibetol acetate (16.9%), 4 –allylphenyl acetate
(9.4%) and 4-allylphenol (7.2%) were the main components, comprising 69.7% of the oil.
The fractionation of the essential oil gave two fractions. Fraction I was rich in eugenol
(71.3%) and fraction II in eugenol (46.4%), chavibetol acetate (19.4%) and 4-allylphenyl
acetate (11.8%).
Vasantha et al. (2012) tested hexane, petroleum ether, chloroform, acetone and
methanol extracts of leaf, stem and tuber of Radrostis foetidissima for qualitative and
quantitative phytochemical analysis. The leaf extracts revealed the presence of flavonoids
and steroids and steroids in all extracts. Flavonoids, tannins triterpenoids, phenols, steroids,
glycosides and cardiac glycosides are present in the chloroform, methanol and acetone
extract while saponin were detected only in methanol and acetone extract.
Kulkarni and Sathe (2013) examined the Hamiltonia suaveolens stem for
phytochemical compounds and GC-MS analysis. The presence of phytochemical
compounds was screened by qualitative method. The results showed the presence of
phytochemical compounds of carbohydrates, terpenoids, steroids, flavonoids, phenols.
Alkaloids were not detected.
2.5. In silico studiesQSAR
Toropov et al. (2009) studied the additive SMILES – Based carcino genicity
models: probabilistic principles in the search for Robust Predictions. Optimal descriptors
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 33
calculated with the simplified molecular input line entry system (SMILES) have been
utilized in modeling of carcinogenicity as continuous values (LogTD50). These descriptors
can be calculated using correlation weights of SMILES attributes calculated by the Monte
Carlo method.
Chaudhry et al. (2010) reported the global QSAR models of skin sensitisers for
regulatory purposes. The new European regulation on chemical safety, REACH
(Registration, Evaluation, Authorisation and Restriction of Chemical Substances) is in the
process of being implemented. Many chemicals used in industry require additional testing
to comply with the REACH regulations. Computational techniques such as QSAR have the
potential to offer an alternative for generating REACH data. The FP6 project CAESAR
was aimed at developing QSAR models for 5 key toxociological end points of which skin
sensitization was one.
Cassano et al. (2010) studied the caesor models for developmental toxicity.
Developmental toxicity is one of the most difficult end point to assess, on account of the
complexcity, lenth and costs of experimenta – following the encouragement of QSAR (In
silico) methods provided in the REACH itself, the CAESAR project the developed several
models. Two QSAR models for developmental toxicity have been developed, using
different statistical / mathematical methods.
Yadav and Khan (2013) studied QSAR, docking and ADMET studies of
camptothecin derivatives as inhibitors of DNA Topoisomerase – 1, quantitative structure –
activity relationship (QSAR) models of camptothecin derivatives against DNA
Topoisomerase – 1 (DNA Topo – 1) were developed by multi linear regression method
using leave one out validation approach.
Docking
Searls (2000) studied the key aspect of drug discovery in the genomic revolution,
contributing to both target discovery and target validation. The pharmaceutical industry has
embraced genomics as a source of drug targets and as a corollary, has recognized that
bioinformatics is crucial to exploiting the data produced on a genome wide scale.
Payne (2004) reported the antibacterial drug discovery has experienced a paradigm
shift from phenotypic screening for antibacterial activity to rational inhibition of
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 34
preselected targets. The pharmaceutical industry embraced genomic information as the
basis for a rational, target directed antibacterial drug discovery strategy to complement the
classic empirical approach.
Computer Aided Drug Design (CADD) is a widely used term that represents
computational tools and resources for the storage, management, analysis and modeling of
compounds. It includes development of digital repositories for the study of chemical
interaction relationships, computer proGrams, for designing compounds with interesting
physicochemical characteristics, as well as tools for systematic assessment of potential lead
candidate before they are synthesized and tested.
Klebe (2006) examined new technologies such as comparative modeling based on
natural structural homologues have emerged and began to be exploited in lead design.
These, together with advances in combinatorial chemistry, high-throughput screening
technologies and computational infrastructures, have rapidly bridged the gap between
theoretical modeling and medicinal chemistry. CADD now plays a critical role in the
search for new molecular entities.
Paramasivan et al. (2006) studied the prediction of 3-dimensional structures of
salivary odorant binding protein-2 of the mosquito Culex quinquefasciatus, the vector of
human lymphatic filariasis. A 3-dimensional model (3D) was developed for the salivary
gland odorant binding protein-2 of the mosquito Culex quinquefasciatus, a major vector of
human lymphatic filariasis. A homology modeling method was used for the prediction of
the structure. Two template proteins were obtained by mGenTHERADER, namely the high
resolution X-ray crystallography structure of a pheromone binding protein (ASPI) of
Apis mellifera L. (IR5R: A) and the aristolochene synthase from Penillium roqueforti
(1DI1: B). The structure of OBP of the mosquito Culex quinquefasciatus resembles the
structure of pheromone-binding protein ASP1 of Apis mellifera L. (IR5R: A).
Babu et al. (2008) examined the synthesis, characterization, anti-microbial activity
and docking studies of novel dispiro-oxindolopyrrolidines. A facile synthesis and
characterization of novel dispiro-oxindolopyrrolidines is delineated. The results of the anti-
microbial studies of novel dispiro-oxindolopyrrolidines are correlated with the docking
calculations performed on proteins S12 from 3OS ribosomal subunit.
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 35
Syed and Leal (2003) examined the nature compounds from natural compounds
from humans and birds, which are detected with extreme sensitivity by olfactory receptor
neurons (ORNs) on the antenne of Culex pipiens quinquefasciatus
Song et al. (2009) studied the improved design and management of data sources,
creation of computer proGrams to generate huge libraries of pharmacologically interesting
compounds, development of new algorithms to assess the potency and selectivity of lead
candidates, and design of predictive tools to identify potential ADME / Tox liabilities.
Yin et al. (2010) observed synthesis and antimicrobial activities of novel peptide
deformylase inhibitors. A new series of N-formylhydroxylamine compounds were
designed, optimized with the Auto Dock 4.0.1. to investigate the interactions between the
target compounds and the amino acid residues of the Escherichia coli. Ni enzyme, and then
synthesized through multi-step sequence starting from diethyl malonate. The structures of
the compounds were characterized on the basis of spectral (FT-IR, IH NMR and Mass)
analysis.
Mao et al. (2010) reported the odorant binding proteins in the female antenna play a
crucial role in sensing oviposition cues. The X-ray crystallography and NMR 3D structures
of Culex quinquefasciatus bound to an oviposition pheromone (5R, 6S)-6-acetoxy-5-
hexadecanolide (MOP).
Kavitha et al. (2011) studied the antimicrobial activity and molecular docking
studies of some novel tetrazole diazepine derivatives. The biological importance of
tetrazole and diazepam moieties, seven novel tetrazole diazepine derivatives (4a, x = -H;
4b, X = -NO2; 4c = (OCHO3)2; 4d = X = Cl; 4e = -Br; 4f = -OH; 4g = C6H5X =
furfuraldehyde) were synthesized.
Hoskeri et al. (2012) examined in-silico drug designing using -sitosterol for
antibacterial activity was investigated by docking -sitosterol molecule with the peptide
deformylase protein and compared with the commercial antibiotic liprofloxaxin. The active
domain of peptide deformylase protein consisted of Gly 45(A), Gln50(A), Leu 91(A), Glu
133(A) as catalytic residues with residues Gly 43(A), Gln 42(A), Arg 97(A), Glu 88(A),
His 132(A), Cys 129(A), Glu 133(A), Gln 50(A), Leu 91(A), His 136(A), Cys 90(A), Iie
44(A), Gly 89(A) in its active pocket. This pocket was docked with -sitosterol and
Results and Discussion
Insilico approaches for characterizing antimicrobial and larvicidal activity of selected plant extracts against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae) 36
ciprofloxycin at the torsional degree of freedom 0.5 units with genetic algorithm resulted
with the inhibition constant as 2.4 x 10-7 for -sitosterol, whereas inhibition constant of
ciprofloxacin was 2.73 x 10-8.
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