mosquito larvicidal activity of leaf and seed extracts of lantana camara on aedes aegypti

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Mosquito larvicidal activity of leaf and seed extracts of Lantana camara on

Aedes aegypti

Keywords: Aedes aegypti, Lantana camara, toxicity, hatchability, development time.

ABSTRACT: Background and Objectives: This paper reports the toxicity of Lantana camara to developmental stages of the yellow fever mosquito, Aedes aegypti. Aqueous extracts of leaf and seed of the plant were also tested for their effect on the hatchability of mosquito egg and age at pupation and emergence. Methods: Different concentrations of aqueous leaf and seed extract were prepared. The data of mortality rate were subjected to finney’s method of probit analysis. The plant was also tested for their effect on the hatchability of mosquito eggs. Results: Percent log LC50 / 24 h values of the leaf and seed extracts of L. camara to IV instar larvae were 2.25 and 2.25 respectively. Percent hatchability of mosquito eggs was remarkably reduced when treated with higher concentration of the toxicants. Extended time of pupation and emergence was observed for the larvae reared in different concentrations of the plant extract. Conclusion: The results suggested that leaf and seed extract of Lantana camera possessed remarkable larvicidal, ovicidal, and prolonged time of pupation and adult emergence against Aedes aegypti.

040-047 | JRAS | 2012 | Vol 1 | No 2

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Journal of Research in

Animal Sciences An International Open Access Online

Research Journal

Authors:

Sathya K2, Mohanraj RS1,

Dhanakkodi B2 .

Institution:

1. Department of Zoology,

Government Arts college

(Autonomous)

Coimbatore - 641 018,

Tamil Nadu, India.

2. Department of Zoology,

Kongunadu Arts and Science

College (Autonomous)

Coimbatore - 641 029,

Tamil Nadu, India.

Corresponding author:

Mohanraj RS.

Email: [email protected]

Web Address: http://ficuspublishers.com/

documents/AS0013.pdf

Dates: Received: 12 Sep 2012 Accepted: 01 Oct 2012 Published: 29 Oct 2012

Article Citation: Sathya K, Mohanraj RS, Dhanakkodi B . Mosquito larvicidal activity of leaf and seed extracts of Lantana camara on Aedes aegypti. Journal of Research in Animal Sciences (2012) 1(2): 040-047

Journal of Research in Animal Sciences

An International Online Open Access

Publication group Original Research

INTRODUCTION

Mosquitoes are the prominent vectors of human

diseases viz., Malaria, yellow fever, dengue, filariasis

and encephalitis (Lymio et al., 1992; Rawlins and Wan,

1995). Among the different species of mosquitoes,

individuals of genus Aedes are considered highly

dangerous because, these show more dependency on

human blood (Scott et al., 1993) and breed in artificial

containers like discarded automobile tires, flower vases,

tin cans, Jars, unused water closets, cisterns and the like

in and around human habitations (Barrera, 1996).

Aedes aegypti is the potential vector of dengue and

yellow fever (Mazzarri and Georghiou, 1995). A severe

out break of dengue with 16 death has been reported in

Tamil Nadu (The Hindu, 2001). In October, 2006, a total

of 5,710 cases were recorded in India. Delhi had the

highest number (1,737) of patients. Tamil Nadu had

307 patients, 103 deaths were also reported (The Hindu,

2006). In 2012 total of 3,000 cases and 50 deaths were

reports in Madurai, Tirunelveli and Kanyakumari

districts (Tamil Nadu) (The Hindu, 2012). Recently out

break of chikungunya has become the talk of the world.

According to central health secretary of India, in 2006,

13 lakh people affected by this disease. In Tamil Nadu

alone 63,000 persons were affected by this disease

(The Hindu, 2006). So, priority must be given to the

control of the mosquitoes for the protection of public

health.

Many control strategies for mosquitoes have

been suggested since ancient times. Physical control is

one of the method by which the physical nature of

environment is changed to reduce breeding of

mosquitoes. Physical control measures include digging

ditches; grading or filling low areas, laying tile etc.

Although these measures are expected to provide

effective mosquito control, the problem is that they are

relatively expensive and must be cooperatively carried

out by landowners, the public and government agencies.

Use of synthetic pesticides is still the most

convenient method all over the world. However,

extensive and indiscriminate use of these pesticides

caused environmental contamination and showed

unwarranted effects on non-target beneficial organism as

well as on human beings (Selvaraj Pandian et al., 1994).

These serious effects arising due to the use of synthetic

chemicals indicate the need for considering alternative

control measures for mosquito.

In recent years, scientists try a variety of

botanical derivatives to eradicate many harmful insect

pests including mosquitoes. Some of the plant products

tested for their insecticidal properties on the medically

and economically important mosquitoes are : Cake

powders of linseed, mustard, castor, mahuana

(Bowry et al., 1984); aqueous, petroleum ether extracts

of Carica papaya, Nerium indicum (Evans and Raj,

1988); steam distilled oils of marigold, Tagetes patula

(Green et al., 1991); methanol and acetone extracts of

Mentha piperita (Selvaraj Pandian et al., 1994);

petroleum ether extracts of Karanja seeds and

Garlic bulbs (Das et al., 1995); oils of Acorus calamus,

Cedrus deodara, Eucalyptus sp., Pongamia glabra,

S e s a m u m i n d i c u m , R i c i n u s c o m m u n i s ,

Madhuca longifolia, Zanthoxylum limonella (fruits)

Citrus aurantifolia (leaf) (Vrushali, 2001;); fruits of

Piper longum (Yang et al., 2002); Crude extract of

Celery, Apium graveolens (Choochote et al., 2004);

aqueous extracts of long pepper (Piper retrofractum)

(Chansang et al., 2005); aqueous leaves extracts of

Lepidagathis alopecuroides (Obomanu et al., 2006) and

leaf extract of Ageratina adenophora (Rajmohan and

Ramaswamy, 2007). Insecticidal activity of neem has

been reported (Mohanraj and Dhanakkodi, 2007, 2008);

aqueous extract of neem leaves (Arunpandiyan, 2011);

methanolic extract of Acalypha alnifolia leaves

(Kovendan et al., 2012).

041 Journal of Research in Animal Sciences (2012) 1(2): 040-047

Sathya et al.,2012

Prohit et al., (1989) demonstrated that plant

products are easily biodegradable, eco-friendly and they

leave no poisonous residues.

Lantana camara (Family: Verbenaceae),

commonly called unnichedi in Tamil is a large shrub

found in tropical and subtropical regions of the world. It

is completely adapted to varied conditions of soil in

India. Chemical constituents of the whole or different

parts of this plant are pentacyclic triterpenoids,

oxosteroid–lanncamarone, triterpene-lantanolic acid and

lantic acid.

Though, the plants are widely distributed in

India, Tamil Nadu in particular, easily accessible and

extensively used for medical purpose, investigations on

their insecticidal properties are scanty, while neem which

was known for its medicinal properties but later on

exploited as pesticide could not be left unnoticed at this

juncture. Very few preliminary studies have been so far

carried out, to evaluate the toxicity and repellency of

Lantana camara (Seyoum et al., 2002; Dua et al., 2003).

It is therefore desirable to study multitargeted

effects of these plants on insect pests, mosquitoes in

particular.

MATERIALS AND METHODS

Animal

Different larval stages of the yellow fever

mosquito, Aedes aegypti were used as experimental

animals in the present study. Eggs of Aedes mosquitoes

were first collected. They were hatched and reared in the

laboratory. The adults were identified and maintained in

cages for generations. The larvae obtained from this

stock were used for different experiments.

Test Compounds

Powdered leaf and seed of plants

Lantana camara which are locally called Unnichedi in

Tamil were used as test compounds in the present study.

Preparation of water extracts of plant parts

Leaf and seed were carefully collected from the

plant and air dried under shade. The dried leaf and seed

were pounded with mortar and pestle to a course powder.

The powder was stored in clean containers until further

use.

A known quantity of the leaf/seed powder of

plant was taken in a container with 200 ml of filtered

unchlorinated water, stirred well and kept for 24 h. The

mixture was again stirred well and filtered through

Whatman No. 1 filter paper. The filtrate was used as

experimental medium in which larvae were reared for

experimental purposes. The amount of powder taken at a

time was in proportion to the concentration of the

medium required. For example, one percent of test

medium of either leaf or seed of Lantana camara was

prepared by dissolving 2 gm of the powder in 200 ml of

water according to the method adopted for preparing of

neem water extracts by Mourier (1997); Mohanraj and

Dhanakkodi (2005).

Test for toxicity of plant extracts

Different concentrations of the test compounds

were prepared using unchlorinated water as described

earlier. Batches of 20 larvae were exposed to 200 ml of a

particular concentration of the test solution. Mortality

rates of larvae were recorded after 24 h. Five or more

concentrations of a test compound recorded between 0 to

100% mortality of larvae at IV instar stage were tested.

Two replicates were done at each concentration. The data

of mortality rate were subjected to Finney's method of

probit analysis as detailed by Regupathy and Dhamu

(1990), toxicity was expressed in terms of LC50 (Median

Lethal Concentration).

Test for the effect of plant extracts on hatchability of

A. aegypti eggs

Fresh eggs of A. aegypti were placed in media

treated with different concentrations (0.5%, 1.0% and

1.5%) of leaf and seed extracts of the test plant and

control as fifty eggs each. Eggs/egg shells were collected

from each container after 96 h from the commencement

of the experiment and the number of hatched and

Journal of Research in Animal Sciences (2012) 1(2): 040-047 042

Sathya et al.,2012

unhatched eggs were recorded. The eggs which found

broken were counted as hatched and those found

complete were considered as unhatched. The time

(96 h) was fixed, because it was demonstrated that the

completion of embryogeny occurs with in four days

(Judson and Gojrati, 1967). The experiment was repeated

thrice. The data were statistically examined using

student ‘t’ test.

Determination of effect of plant (leaf / seed) extracts

on the development time of A. aegypti

Three batches of first instar larvae as 50 each of

A. aegypti, were introduced into the media of particular

concentration of the test extracts. They were maintained

with fish food ad libitum until all surviving larvae

pupated. The pupae were also allowed to emerge as

adults. The medium was replaced with freshly prepared

test solutions every alternate day. Number of larvae

completed ecdysis i.e., conversion from one instar stage

to the subsequent one, was recorded every 24 h till

emergence. The daily record of mortality among the

larvae reared in each concentration of the plant extract

and control was maintained. Accordingly the percentage

of larvae pupated and emerged into adults and mean time

required for pupation and emergence was calculated

(Sagar and Sehgal, 1997). The data were statistically

examined using student ‘t’ test.

RESULTS AND DISCUSSION

Toxicity of Lantana camara to different larval stages

of A. aegypti

Log LC50 / 24 h values of leaf extracts of

Lantana camara to the IV instar larvae of the mosquito

was 2.25 (LL-2.22 and UL - 2.28) and that of the seed

extract it was 2.25 (LL-2.19 and UL-2.31), respectively.

(Fig 1and 2).

Sathya et al.,2012

Concentration No. of eggs introduced No. of eggs hatched % hatch Per cent reduction over control

Control 50 40.33 ± 1.25 80.66 -

0.5% 50 40 ± 0.82 80 0.82

1% 50 30 ± 0.82 60 25.6

1.5% 50 18.33 ± 1.25 36.66* 54.55

* Significantly different (P < 0.01) from that of control

Table 1 Change in the hatchability of Aedes aegypti eggs treated with

different concentrations of leaf extract of Lantana camara and control.


0.0 0.5 1.0 1.5 2.0 2.5 3.0

1

2

3

4

5

6

7

8

9

Pro

bit o

f M

ort

alit

y

Log dose

Fig 1 Probit regression line for the toxicity of

Lantana camara (leaf) on fourth instar larvae of

Aedes aegypti

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5

1

2

3

4

5

6

7

Pro

bit o

f M

ort

alit

y

Log dose

Fig 2 Probit regression line for the toxicity of

Lantana camara (seed) on fourth instar larvae of

Aedes aegypti

Fig 1 r value =0.478 slope=24 Fig. r value=0.758 slope=38

Effect of the plant extracts on hatchability of

A. aegypti eggs

Percent hatch of the mosquito eggs in control

medium was 80.66. This was reduced to 36.66 and 8 for

the eggs exposed to 1.5% of leaf and seed extracts of

L. camara (Tables 1and 2).

Effect of Leaf and seed extracts of Lantana camara on

mean age at pupation and emergence of A. aegypti

Larvae of A. aegypti reared in control medium

pupated at 132.67 h (5.5 days) and adults emerged at

184.67 h (7.6 days). Leaf and seed extracts of L. camara

prolonged the period of pupation and adult emergence.

Mosquito larvae reared in the leaf extract (1.5%) of

L. camara completed pupation in about 176 h

(7 days) and those developed in a similar concentration

of seed extract attained pupation at 196.3 h (8.2 days)

(Fig. 3 and 4). This was significantly different (p < 0.01)

from that of the control.

In the present study, leaf & seed extracts of

L.camara showed notable toxic effect on fourth instar

larvae of A.. aegypti. The log median lethal concentration

(Log LC50 / 24 h) of leaf and seed extracts of L. camara

was 2.25 respectively.

Though exact comparisons are not available, due

to differences in the methods of the experimental set up,

reports of Dua et al., (1996) and Seyoum et.al., 2002 on

insecticidal properties of L. camara may be quoted in

support of the present observation.

Leaf and seed extracts of L. camara as observed

in the present study, showed ovicidal effect on the eggs

of A. aegypti. Percent hatching of eggs placed in control

medium (filtered tap water) was 80.66. This was reduced

to a considerable level of 36.66 and 8 when treated with

1.5% of the plant extracts.

Reduction in the hatching of eggs from

lepidopteran and hymenopteran insects on treatment with

aqueous extracts of plant derivatives such as black

pepper, neem seed and garlic bulb was recorded by

Ekesi (2000). It was opined that chemicals synonym to

insect growth regulators diffuse into eggs and effect vital


Sathya et al.,2012

Concentration No. of eggs introduced No. of eggs hatched % hatch Per cent reduction over control

Control 50 40.33 ± 1.25 80.66 -

0.5% 50 35 ± 0.82 70 13.22

1% 50 7 ± 0.82 14 82.6

1.5% 50 4 ± 0.82 8* 90.1

Table 2 Change in the hatchability of Aedes aegypti eggs treated with

different concentrations of seed extract of Lantana camara and control.

* Significantly different (P < 0.01) from that of control

0

50

100

150

200

250

300

Control 0.50% 1% 1.50%

Ho

urs

0

2

4

6

8

10

12

Day

s

Mean age at pupation (hours) Mean age at Emergence (hours)

Mean age at pupation (Days) Mean age at Emergence (Days)

Fig 3 Change in development time of Aedes aegypti

larvae reared in different concentrations of leaf

extract of Lantana camara and control

0

50

100

150

200

250

300

Control 0.50% 1% 1.50%

Ho

urs

0

2

4

6

8

10

12

Day

s

Mean age at pupation (hours) Mean age at Emergence (hours)

Mean age at pupation (Days) Mean age at Emergence (Days)

Fig 4 Change in development time of Aedes aegypti

larvae reared in different concentrations of seed

extract of Lantana camara and controls

physiological and biochemical processes associated with

embryonic development, there by inhibiting eclosion of

eggs (Boradbent and Pree, 1984).

Crude water extracts obtained from leaf and seed

of L. camara were found to prolong the larval period of

A. aegypti and thus delay pupation and emergence. Mean

age at pupation for the larvae reared in control medium

was 132.67 h. This was extended to 176, 196.3 h in

larvae exposed to the media (1.5%) of leaf and seed

extracts of L. camara, respectively. The prolonged

life-span of aquatic larvae was described as a

consequence of low feeding rate (Pandian and

Madhavan, 1974).

CONCLUSION

The finding of the present investigation revealed

that aqueous leaf and seed extract has good toxic,

ovicidal and prolong the larval period of A. aegypti and

thus delay pupation and emergence.

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