Evaluation of pesticidal activities of Lantana camara against Tribolium confusum and its

comparison with synthetic insecticide Cypermethrin

Haroon Ur Rashid1, Seema Tahir 1, Muhammad Rizwan Amjad2, Abdul Khaliq2, Adnan Amjad3,
Maheen Fatima2, Muhammad Aqil Qureshi2, Sehrish Kanwal4, Muhammad Farhan somro2
1
Department of Zoology, Karachi University Campus, Karachi, Pakistan
2
Pest Warning and Quality Control of pesticides Punjab, Lahore, Pakistan
3
Instiute of Food Science and Nutrition, Bahauddin Zakariya University Multan
4
Department of entomology PMAS Arid University of Agriculture, Pakistan

Corresponding Author
Muhammad Rizwan Amjad
rizwanamjad41@gmail.com

Abstract

Tribolium confusum is a notorious insect pest that destroyed a huge number of stored
commodities at the storage level. There is a need to find a naval, easily degradable and eco-
friendly replacement of conventional insecticides to control insect pest in stored grains. The
toxicity, residual toxicity, repellency and progeny effects of the plant extract Lantana camara
were compared with synthetic insecticide cypermethrin against T. confusum. Five different
concentrations of plant extracts (1%, 1.2%, 1.4%, 1.6%, 1.8%) were checked in each
experiment. Synthetic insecticide was applied at concentrations (0.0625%, 0.0313%,
0.0156%, 0.0078%, 0.0039%) in each bioassay. In toxicity bioassay, the LC 50 of
cypermethrin and L. camara were calculated to be 6.1µg/cm 2 and 298.8 µg/cm2 after 24 and
48 hrs, respectively. Maximum mortalities were recorded, 92.8% and 64.2% for cypermethrin
and L. camara, respectively, against T. confusum. Residual toxicity of cypermethrin (12.5
µg/cm2) produced percent mortalities 86.21%, 89.66%, 48.28%, 31.04% and 10.35% while
for L. camara at dose of 358.5 µg/cm2 were 44.83%, 48.28%, 41.38%, 31.04% , 13.79%
respectively after 1, 2, 3, 4 and 5 days. In the repellency test, cypermethrin and L. camara
produced percent repellency of 73.3% and 66.7% respectively after 5 hrs. In contrast, the
fecundity effect at LC50 dose inhibited 77.16% and 67.8% insect emergence after 42 days.
Current research showed that plant extract plays an important role in the management of T.
confusum. The tested plant effectively controlled the pest and suggested using against stored
products insect pest as a part of integrated pest management.

Keywords: T. confusum, L. camara, Mortality, Repellency, Fecundity

1.Introduction

The damage of stored grain, agricultural products and seeds from insect pests has been
a huge problem from the onset of agriculture development (Mahfuz and Khanam, 2007).
Wheat constitutes 80% of food among the cereals in Pakistan, is very sensitive to insects,
rodents and fungal attacks (Haq et al., 2005). During storage, a massive quantity of the
cereals destroyed in temperate countries; the losses ranged from 5-10%  whereas, in tropical
countries, the losses reach from 10-40% annually either qualitatively or quantitatively, only
owing to different stored product insect pests (Tripathi et al., 2009; Ahmedani et al., 2011;

1
Komaki et al., 2017). Food security is mainly dependent on some major mitigation like the
proper storage space, sanitation, quality assurance and handling and protection of stored
grains materials all around the globe (Ahmed, 1983, Belluco et al., 2013). Stored grain pests
reduce the nutritive and market values of food (Oboh et al., 2017; Ali, 2013). About 600
beetles, 70 months and 355 mites species attack agricultural products at storage level
(Rajendran, 2002). Nation backwardness is directly proportioned to their stored product
losses (Ashfaq et al., 2001). As agriculture sector is considered the backbone of Pakistan's
economy, which provides 20.9 GDP, called national GDP (Chandio et al., 2016). Out of 80
million hectares of land of the country, about 22 million hectares are cultivated for
agriculture (Aslam, 2016). Major agricultural crops are cotton, rice, maize, sugarcane and
wheat (Ahmed et al., 2017).

In Pakistan, losses of stored product commodities due to insect pest infestation have
been reported about 10-15% (Jilani, 1981). T. confusum, Sitophilus oryzae, Rhyzopertha
dominica, T. castaneum and Trogoderma granarium are major insect pests of wheat in
Pakistan (Prikhodko, 2013). Among these stored product pests, T. confusum is a major insect
pest also called confused flour beetle, is a cosmopolitan stored grain insect pest (Wenda-
Piesik et al., 2017). It is one of the notorious primary pests of stored grains (Hamzavi and
Moharramipour, 2017). T. confusum adult is reddish-brown in color, 4 - 4.5 mm in length and
1.0 - 1.2 mm in width, while morphologically, males and females are indistinguishable under
a light microscope. Setiferous patches on adult male legs are observed under the electron
microscope (Zohry, 2017).

The use of synthetic insecticides for the control of stored product pests penetrates and
sometimes becomes more toxic (Ogendo et al., 2003). Some synthetic insecticides reported
having mutagenic and carcinogenic effects (Lingampally et al., 2013). Cypermethrin is a
member of pyrethroid; their insecticidal potential was reported from 1970. Cypermethrin is
abundantly used for the management of T. confusum (Coleoptera) and other pests of
ornamental and store cereals (Pankaj et al., 2016). These are broadly utilized in homes,
agriculture, public health, horticulture, textiles protection and forestry (Lin et al., 2011;
Zhang et al., 2011). In contrast, T. spp were found resistive to cypermethrin. It disrupts the
neuro-endocrine and biological functions of insects (Rowland, 1995). Imidacloprid is
neurotoxic in action and belongs to the class of neonicotinoids. It is used for the treatment of
seed coats and has significant control results against various pests (Zhang et al., 2011).
Insects of different orders, Lepidoptera, Coleoptera, Homoptera and Diptera, have been

2
controlled excellently with imidacloprid (Wakil and Schmitt, 2015). Due to the adverse
ecological and human health effects, researchers divert their attention toward alternative safe
methods to control pests (Hanif et al., 2016). Therefore, plant-origin natural compounds have
many advantages over synthetic pesticides and suggest an important perspective for pest
control programs (Kachhwaha et al., 2015). Therefore the main objectives of the current
study are to determine the efficacy (repellency and fecundity) of L. camara plant extract in
comparison with synthetic insecticides Cypermethrin against test insects. Also, determine the
lethal dose (LD50) and its residual toxicity against stored grain insect pests.

2.Materials and Methods

The present investigation was carried out at the Department of Zoology, Biological
Research Centre (B.R.C), Southern Agricultural Research Center (S.A.R.C), Pakistan
agriculture research center (P.A.R.C), University of Karachi.

2.1Insect rearing

T. confusum was mass-reared for experiments in the insectary of SARC. The culture
was maintained for six months. The susceptible test insect was obtained from the existed
culture of T. confusum at GSRL, SARC. Twenty grams of wheat flour was taken in a glass jar
and 1gram of yeast (1:20) was added to it, then thirty adult insects were put in each jar and
covered with muslin cloths and rubber bands. The jars were kept in an incubator at a relative
humidity of 65+5% and a temperature of 30+2C˚. The insects were left 10 days for
oviposition in the jars. After then the adults were shifted to new jars and the procedure was
continued for 4 months to obtain a homogenous population (same sized insect) for toxicity
bioassay. Eight days adult insects (mixed sex) were used in bioassays.

2.2Synthetic insecticides and plant extracts

Two plants extract, i.e., L. camara was used in the present study. While cypermethrin,
10% EC was used for positive control or comparison.

2.3Insecticide

Cypermethrin (10% EC) commercial-grade insecticide was obtained from the

Department of Plant Protection, Malir Halt, Karachi. Cypermethrin 1.25 gram was diluted in
the final volume of 100ml solvent (ethanol 50%) and 1 % stock solution (W/V) was
prepared.by using the above solvent, from the stock solution five different serial dilutions i-e
0.0625%, 0.0313%, 0.0156%, 0.0078% and 0.0039% prepared These concentrations were

3
converted into per unit area dose i-e 12.5 µg/cm2, 6.25 µg/cm2, 3.13 µg/cm2, 1.6 µg/cm2, and
0.8 µg/cm2, respectively.

2.4Collection of plants

L. camara was collected from the Department of Geography, University Of Karachi.

Both the plants were authenticated and identified by the Center of Plant Conservation Botany
Herbarium, University of Karachi.

2.5Preparation of plant extracts

2.5.1Leaves extract of L. camara

Fresh leaves of L. camara 2kgs were rinsed with tap water 3-4 times, air-dried in the
shade (to avoid destruction of the active component or alkaloid) for 15 days. Air-dried leaves
were macerated with an electric grinder. 100gm powder was soaked into 500ml of ethanol
50%. The sample was kept in an electric shaker of 120r/min and 28 C˚ for 7 days. After one
week, the sample was filtered 2 times from filter paper and the filtrates were run by a rotary
evaporator. All the solvent was evaporated and the gummy substance remained in the flask.
Three grams of plant compounds were dissolved into the final volume of 100ml ethanol
(50%) to prepare a 3% stock solution from that five dilutions of 1%, 1.2%, 1.4%, 1.6% and
1.8% were prepared, which was then converted into per unit area i.e 199.2 µg/cm 2, 239
µg/cm2, 278.9 µg/cm2, 318.7 µg/cm2 and 358.5 µg/cm2, respectively.

2.6Bioassay/Experiments

2.6.1Toxicity

Toxicity bioassay was conducted on newly emerged adults of T. confusum by filter

impregnation method adopted by (Arif et al., 2013). For this purpose, 72 Petri dishes of 8 cm
diameter were rinsed, sterilized and dried in the oven. All Petri dishes were equal in size and
shape. The diameter of each Petri dish was 8 cm. The filter paper was cut according to the
diameter of the petri dish and placed in all Petri dishes, impregnated by different doses of
insecticides and plant extracts. Synthetic insecticide dilutions i-e 0.0625%, 0.0313%,
0.0156%, 0.0078% and 0.0039% and plant extract i-e 1%, 1.2%, 1.4%, 1.6% and 1.8% used
in Petri dishes along with reference /control. The experiments were replicated three times.
The 1ml dose was applied with the help of a pipette. Filter papers were air-dried for 15
minutes. Ten insects of the same size and age were kept to each petri dish. Insect mortality
was observed and recorded after 24, 48 and 72 hours. Each experiment was repeated four

4
times. LD50 of each dose was recorded. Mortality was corrected by the Abbot formula
(Abbot, 1925). The data were analyzed statistically and the regression model was developed
by Microsoft excel 2007 and standard error, standard deviation and confidence limit 95 % of
data were calculated.

2.6.2Residual toxicity

For residual bioassay, 8cm diameter Petri dishes rinsed and sterilized were used and the
filter papers were impregnated with respective insecticides dilutions and plant extracts then
dried for 15 minutes. Ten adult insects of the same age and size were kept to each Petri plate.
Mortality was observed after 24 hours and all insects were removed after 24 hours and fresh
10 adult insects were added to each Petri plate. Data was taken after 48 hours. All the insects
were removed after 48 hours and fresh insects were added. The experiment was continued till
the mortality reached up to 0%. The insecticide dilutions were (0.0625%, 0.0313%, 0.0156%,
0.0078% and 0.0039%) then converted into per unit area dose i-e 12.5 µg/cm 2, 6.25 µg/cm2,
3.13 µg/cm2, 1.6 µg/cm2 and 0.8 µg/cm2, respectively. While plant extract dilutions were 1%,
1.2%, 1.4% 1.6% and 1.8% converted into per unit area i.e 199.2 µg/cm2, 239 µg/cm2, 278.9
µg/cm2, 318.7 µg/cm2 and 358.5 µg/cm2, respectively.

2.6.3Repellency test

Eight cm diameter of filter paper was sliced into half and utilized for the experiment.
Insecticide dilutions i-e 0.0625%, 0.0313%, 0.0156%, 0.0078% and 0.0039% concentrations
converted into per unit area dose i-e 12.5 µg/cm2, 6.25 µg/cm2, 3.13 µg/cm2, 1.6 µg/cm2 and
0.8 µg/cm2, respectively while plant extracts 1%, 1.2%, 1.4%, 1.6% and 1.8% concentrations
converted into per unit area i.e 199.2 µg/cm2, 239 µg/cm2,278.9 µg/cm2, 318.7 µg/cm2 and
358.5 µg/cm2, respectively were used in experiments. Half of the filter paper was treated with
0.5 ml of concentration by pipette, while half was left as control. Both parts were rejoined
with squash tape and placed in Petri plates. Ten adult insects were released at the center of
each petri dish. A small amount of diet was provided to each side of the filter paper. Data was
taken after 1 hour and 5 hours. Repellency percentage was calculated by the formula adopted
by (Talukdar and Howse, 1995). The given formula was adopted for calculation

PR ( % ) =(Nt − Nc)/(Nt + Nc)×100

PR (%) = percentage repellency, Nt =number of insects in control half, Nc = number of

insects in treated half

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2.6.4Fecundity

From the mass culture of T. confusum 5th instar, newly emerged larvae were shifted to
media treated with LD50 doses of insecticide (cypermethrin) and plant extracts (L. camara)
with a control (untreated) batch. The male and female sex of T. was separated by the method
adopted by (Halstead, 1963). After adult emergence, 10 days old insects from treated media
were paired. Five pairs (10 insects) were shifted to Petri plates with 5 grams of food; control
was maintained from untreated media. The adults were left 14 days to oviposit in the Petri
plates. Observations were made after 14, 28 and 42 days as adopted by (Shikdar and
shahjahan, 2011).

Formula: Percentage = number of adults/number of eggs x 100

For egg inhibition and emergence, the following formula was used:

Control-treated/control x 100

Statistical analysis
Toxicity: From mean mortality = average mortality /total insects x100
Conversion of ml to µg/cm2

µg/cm2=concentration /100 v/area of petri dish x 106

Abbot’s formula = %mortality-control mortality /100-control mortality x 100.

For serial dilution, Charles's equation was used:

C1V1=C2V2

In the above equation:

C1= stand for initial concentration of solute

V1= stand for initial volume of solute

C2= Final concentration of solvent

V2= final volume of solvent.

Results

Toxicity

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 Cypermethrin toxicity against T. confusum was determined by the filter impregnated
method. Mean mortalities were calculated after 24 hrs duration as 13.7%, 20.6%, 37.9%, 55.1
% and 82.7 % at per unit area dose of 12.5µg/cm 2, 6.25 µg/cm2, 3.13 µg, 1.6 µg/cm2 and 0.8
µg/cm2, respectively. The LC50 value calculated was 6.1 μg/cm2 (Table 1 and Fig. 1).
Cypermethrin toxicity after 48 hrs by the filter impregnated method against T.
confusum was calculated. Mean mortalities were 17.2 %, 24.1%, 44.8%, 58.6% and 86.2% at
per unit area dose of 12.5µg/cm2, 6.25 µg/cm2, 3.13 µg/cm2, 1.6 µg/cm2 and 0.8 µg/cm2,
respectively. The LC50 value calculated was 5.37 μg/cm2 (Table 2 and Fig. 2).
Cypermethrin toxicity after 72 hrs by the filter impregnated method against T.
confusum was calculated. Mean mortalities were 17.8%, 28.5%, 50%, 57.1% and 92.8% at
per unit area dose of 12.5µg/cm2, 6.25 µg/cm2, 3.13 µg/cm2, 1.6 µg/cm2 and 0.8 µg/cm2,
respectively. The LC50 value calculated was 4.78 μg/cm2 (Table 3 and Fig. 3).
L. camara toxicity against T. confusum was determined by filter impregnated method.
Mean mortalities were calculated after 24 hrs duration as 10.3%, 17.2%, 24.1%, 27.5% and
44.8% at per unit area dose of 199.2 µg/cm 2, 239 µg/cm2, 278.9 µg/cm2, 318.7 µg/cm2 and
358.5 µg/cm2, respectively. The LC50 value calculated was 398.8 μg/cm2 (Table 4 and Fig. 4).
L. camara toxicity after 48 hrs by filter impregnated method against T. confusum was
calculated. Mean mortalities were 27.5%, 31%, 37.9%, 55.1% and 58.6% at per unit area
dose of 199.2 µg/cm2, 239 µg/cm2, 278.9 µg/cm2, 318.7 µg/cm2 and 358.5 µg/cm2,
respectively. The LC50 value calculated was 298.8 μg/cm2 (Table 5 and Fig. 5).
L. camara toxicity after 72 hrs by filter impregnated method against T. confusum was
calculated. Mean mortalities were 30%, 43.3%, 46.6%, 53.3% and 66.67% at per unit area
dose of 199.2 µg/cm2, 239 µg/cm2, 278.9 µg/cm2, 318.7 µg/cm2 and 358.5 µg/cm2,
respectively. The LC50 value calculated was 318.7 μg/cm2 (Table 6 and Fig. 6).

Residual toxicity

Cypermethrin residual toxicity against T. confusum was observed. After 1 day of

treatment residual effects of cypermethrin caused mortalities of 17.24%, 31.04%, 58.62%,
75.87% and 86.21% on concentrations of per unit area doses of 0.8 µg/cm 2, 1.6 µg/cm2, 3.13
µg/cm2, 6.25 µg/cm2 and 12.5 µg/cm2respectively. On next second day of insects treatment
percentage mortalities was noted to be 24.14%, 37.93%, 65.52%, 79.32% and 89.66 % on
concentrations of per unit area doses of 0.8 µg/cm 2, 1.6 µg/cm2, 3.13 µg/cm2, 6.25 µg/cm2
and 12.5µg/cm2, respectively. After second day percent mortalities increased gradually and
were found 89.66% at 12.5µg/cm2. Residual effects decreased gradually and on 3 rd day the

7
value of % mortality was observed 48.28 % at highest concentration of 12.5 µg/cm 2 while the
remaining concentrations created percentage mortalities 10.35%, 17.24%, 31.04% and
34.49% at per unit area doses of 0.8 µg/cm 2, 1.6 µg/cm2, 3.13 µg/cm2, and 6.25 µg/cm2,
respectively. On next 4th day the percent mortalities reduced gradually and were found
10.35%, 13.79%, 20.69%, 20.69% and 31.04% at concentrations of per unit area doses of 0.8
µg/cm2, 1.6 µg/cm2, 3.13 µg/cm2, 6.25 µg/cm2 and 12.5µg/cm2, respectively. While after 5th
day percent mortality reduced to 6.9%, 10.35%, 10.35%, 13.8% and 10.35% at
concentrations of per unit area doses of 0.8 µg/cm 2, 1.6 µg/cm2, 3.13 µg/cm2, 6.25 µg/cm2
and 12.5µg/cm2, respectively (Table 7 and Fig. 7).

L. camara residual toxicity against T. confusum was observed. After 1 day of

treatment residual effects of L. camara caused mortalities of 10.35%, 17.24%, 24.14%, 27.59
and 44.83% on concentrations per unit area dose of 199.2 µg/cm 2, 239 µg/cm2, 278.9
µg/cm2, 318.7 µg/cm2 and 358.5 µg/cm2, respectively. On second day mortalities increased
and noted to be 17.24%, 27.59%, 31.04%, 37.93% and 48.28% at per unit area dose of 199.2
µg/cm2, 239 µg/cm2, 278.9 µg/cm2, 318.7 µg/cm2 and 358.5 µg/cm2, respectively. On next 3rd
day highest dose of 358.5 µg/cm2 of L. camara produced 41.38% mortality while the
remaining concentration created mortalities 31.04%, 34.49%, 37.93%, 37.93% at per unit
area dose of 199.2 µg/cm 2, 239 µg/cm2, 278.9 µg/cm2, 318.7 µg/cm2respectively, due to the
residual effects. The mortality reduced gradually on 4 th day and observed to be 17.24% for
three doses 199.2 µg/cm2, 239 µg/cm2and 278.9 µg/cm2, whereas the remaining two doses
318.7 µg/cm2 and 358.5 µg/cm2 produced 27.59% and 31.04% mortalities, respectively. The
lowest residual effects on 5th day observed to be 3.45% at 239 µg/cm2dose, 6.9 % mortality
on 278.9 µg/cm2 and 318.7 µg/cm2 whereas 13.79% observed at highest dose of 358.5 µg/cm2
(Table 8 and Fig. 8).

Repellency

Repellency of T. confusum towards cypermethrin enhanced with the duration of time

exposure. The highest concentration (12.5µg/cm2) of cypermethrin after 1 hour caused a
maximum of 46.6% repellency while a minimum 13.3 % repellency was observed by the
lowest concentration of (0.8 µg/cm2) whereas the highest concentration (12.5 µg/cm2) of
cypermethrin after 5 hours produced maximum 73.3% repellency and the lowest (0.8 µg/cm 2)
concentration produced minimum 20% repellency (Table 9 and Fig. 9).
Repellency of T. confusum towards L. camara enhanced with the duration of time
exposure. The highest concentration (358.5 µg/cm2) of L camara after 1 hour caused a

8
maximum of 26.6% repellency while a minimum of 6.7 % repellency was observed by the
lowest concentration of (199.2 µg/cm2) whereas the highest concentration (358.5 µg/cm2) of
L. camara after 5 hours produced maximum 66.6% repellency and the lowest (199.2µg/cm 2)
concentration produced minimum 13.3% repellency (Table 10 and Fig. 10).

Fecundity

For the determination of physiological effects on egg production, the emergence of

adult T. confusum was treated with LC50 doses of the respective insecticides and plant
extracts. The fecundity effect of cypermethrin was noticed after 14 days of initial treatment.
LC50 5.37 µg/cm2 of cypermethrin against T. confusum by diet incorporated technique. The
mean number of eggs and mean emergence in both treated and control were observed. In this
bioassay, in treated dishes, percent egg lying inhibition was (40%), percent emergence
inhibition (47.5%) whereas, average emergence number observed in both control and treated
dishes was found 81.56% and 71.11%, respectively. After the duration of 28 days, percent
egg lying inhibition was (58%), percent emergence inhibition (75%) while average
emergence number observed in both control and treated dishes was found 88.89% and
53.50%, respectively. Observations after the duration of 42 days T. confusum percent egg
lying inhibition (72.36%), percent emergence inhibition was (77.16%), and average insect
emergence number observed in both control and treated dishes was found 70.06% and
57.73%, respectively (Table 11 and Fig. 11).

Fecundity of T. confusum was observed after the application of LC 50 389.8 µg/cm2 of L.

camara extract by diet incorporated technique. The percentage of eggs lying inhibition in
treated dishes was (46.4%), the percentage of emergence inhibition (55.5%) after 14 days,
while the average amount of emergence observed in both control and treatment dishes was
87.06 percent and 72.18 percent, respectively. After the duration of 28 days, percent egg
lying inhibition was (49%), percent emergence inhibition (56.6%), while average emergence
number observed in both control and treated dishes was found 93.08% and 80.24%,
respectively. Observations after a duration of 42 days, T. confusum percent egg lying
inhibition (66.5%), percent emergence inhibition was (67.8 %), and average insect emergence
number observed in both control and treated dishes was found 85.35% and 82.08%,
respectively (Table 12 and Fig. 12).
Table 1. Toxicity of cypermethrin against T. confusum after a period of 24 hours by the paper
impregnated method. LC50 value calculated was 6.1 µg/cm2 (Mortality was corrected by abbot
formula).

9
 S.N %Conc. Mean % Confidence limit
Conc. /unit S.D S.E
o used Mortality at 95%
area (µg/cm2)
1 Control -- 00 -- -- --
2 0.0039% 0.8 13.7% 1.15 0.67 1.872-1.461
3 0.0078% 1.6 20.6% 1.53 0.88 1.539-3.128
4 0.0156% 3.13 37.9% 1.00 0.58 2.205-5.794
5 0.0313% 6.25 55.2% 2.08 1.20 2.872-8.461
6 0.0625% 12.5 82.8% 1.53 0.88 4.539-12.127
Table 2. Toxicity of cypermethrin against T. confusum after a period of 48 hours by paper
imprengnated method. LC50 value calculated was 5.37 µg/cm2.

S.N %Conc. Conc. /unit Mean % Confidence limit

S.D S.E
o Used area (µg/cm2 Mortality at 95%
1 Control -- 00 -- -- --
2 0.0039% 0.8 17.2 1.00 0.578 2.205-1.794
3 0.0078% 1.6 24.1 1.155 0.667 1.872-3.461
4 0.0156% 3.13 44.8 0.577 0.333 2.872-6.461
5 0.0313% 6.25 58.6 1.732 1.000 3.205-8.794
6 0.0625% 12.5 86.2 1.155 0.666 4.872-12.461

Table 3. Toxicity of cypermethrin against T. confusum after a period of 72 hours by the paper
impregnated method. LC50 value calculated was 4.78 µg/cm2.

S.N %Conc. Conc./unit Mean % Confidence limit

S.D S.E
o used area(µg /cm2 Mortality at 95%
1 Control -- 00 -- -- --
2 0.0039% 0.8 17.8 1.154 0.666 2.538-2.127
3 0.0078% 1.6 28.5 1.154 0.666 2.538-4.127
4 0.0156% 3.13 50.0 1.154 0.666 3.538-7.127
5 0.0313% 6.25 57.1 1.732 1.000 3.205-8.794
6 0.0625% 12.5 92.8 0.57 0.333 5.538-13.127

Table 4. Toxicity of L. camara against T. confusum after a period of 24 hours by the paper
impregnated method. LC50 value calculated was 398.8 µg/cm2.

%Conc. Conc./unit Mean % Confidence limit

S.No S.D S.E
used area(µg/cm2) Mortality at 95%
1 Control -- 00 -- -- --
2 1.000% 199.2 10.3 0.58 0.33 1.538-1.127
3 1.200% 239.0 17.2 1.00 0.58 1.205-2.794
4 1.400% 278.9 24.1 0.57 0.33 0.872-4.461
5 1.600% 318.7 27.5 1.00 0.57 0.205-5.794
6 1.800% 358.5 44.8 0.58 0.33 0.872-8.461

10
Table 5. Toxicity of L. camara against T. confusum after a period of 48 hours by the paper
impregnated method. LC50 value calculated was 298.8 µg/cm2.

%Conc. Conc./unit Mean % Confidence limit

S.No S.D S.E
used area(µg/cm2) Mortality at 95%
1 Control -- 00 -- -- --
2 1.000% 199.2 27.5 1.00 0.58 3.205-2.794
3 1.200% 239.0 31.0 0.58 0.33 2.538-4.127
4 1.400% 278.9 37.9 1.00 0.57 2.2054-5.794
5 1.600% 318.7 55.2 1.15 0.67 2.872-8.461
6 1.800% 358.5 58.6 1.00 0.57 2.205-9.794

11
Table 6. Toxicity of L. camara against T. confusum after a period of 72 hours by the paper
impregnated method. LC50 value calculated was 318.7 µg/cm2.

%Conc. Conc./unit Mean % Confidence limit

S.No S.D S.E
used area(µg/cm2) Mortality at 95%
1 Control -- 00 -- -- --
2 1.000% 199.2 25.00 2.645 1.527 3.205-2.794
3 1.200% 239.0 39.2 1.527 0.881 3.538-5.127
4 1.400% 278.9 42.9 0.577 0.333 2.872-6.461
5 1.600% 318.7 50.00 3.214 1.855 2.538-8.127
6 1.800% 358.5 64.2 2.516 1.452 2.872-10.461

Table 7. Residual toxicity of cypermethrin against T. confusum after 1, 2, 3, 4 and 5 days of

different doses of treatment.
Residual mortality after a range of
Conc.
S.No %Conc. intervals
(µg/cm2)
used 1 day 2days 3days 4days 5days
1 Control …….. 000 000 000 000 000
2 0.0039% 0.8 17.24 24.14 10.35 10.35 6.90
3 0.0078% 1.6 31.04 37.93 17.24 13.79 10.35
4 0.0156%, 3.13 58.62 65.52 31.04 20.69 10.35
5 0.0313%, 6.25 75.87 79.32 34.49 20.69 13.79
6 0.0625% 12.5 86.21 89.66 48.28 31.04 10.35

Table 8. Residual toxicity of L. camara against T. confusum after 1, 2, 3, 4 and 5 days of

different doses of treatment.
Residual mortality after a range of
Conc.
S.No %Conc. intervals
(µg/cm2)
used 1 day 2days 3days 4days 5days
1 Control …….. 000 000 000 000 000
2 1.00% 199.2 10.35 17.24 31.04 17.24 0.00
3 1.20% 239.0 17.24 27.59 34.49 17.24 3.45
4 1.40% 278.9 24.14 31.04 37.93 17.24 6.90
5 1.60% 318.7 27.59 37.93 37.93 27.59 6.90
6 1.80% 358.5 44.83 48.28 41.38 31.04 13.79

Table 9. Cypermethrin repellency against T. confusum after treatment of 1 and 5 hours on

different doses.

S.NO %Conc. used Conc /unit area 1 Hour After 5 Hour

12
 (µg/cm2) After
1 0.0039% 0.8 13.3% 20.0%
2 0.0078% 1.6 20.0% 26.6%
3 0.0156%, 3.13 26.6% 33.3%
4 0.0313%, 6.25 40.0% 40.0%
5 0.0625% 12.5 46.6% 73.3%

Table 10. L. camara repellency against T. confusum after treatment of 1 and 5 hours on
different doses.

Conc /unit area 5 Hour

S.NO %Conc. used 1 Hour After
(µg/cm2) After
1 1% 199.2 6.7% 13.3%
2 1.2% 239.0 13.3% 20.0%
3 1.4% 278.9 26.7% 26.7%
4 1.6% 318.7 20.0% 40.0%
5 1.8% 358.5 26.7% 66.7%

Table 11. Fecundity effects of cypermethrin on T. confusum after 14, 28 and 42 days on LD50
doses.

S. Average Average Average Egg-laying Emergence

Days Treatment
No eggs adult emergence inhibition % inhibition %
1 After control 150.00 122.33 81.56 0 0
14
2 treated 90.00 64.00 71.11 40 47.5
days
1 After control 264.00 234.67 88.89 0 0
28
2 treated 109.67 58.67 53.50 58 75
days
1 After control 825.00 578.00 70.06 0 0
42
2 treated 228.67 132.00 57.73 72.36 77.16
days

Table 12. Fecundity effects of L. camara on T. confusum after 14, 28 and 42 days on LD50
dose.

S.N Day Treatmen Averag Averag Average Egg-laying Emergence

O s t e eggs e adult emergenc inhibition inhibition

13
 e % %
1 After Control 237.00 206.33 87.06 0 0
14
2 Treated 127.00 91.67 72.18 46.4 55.5
days
1 After Control 549.33 511.33 93.08 0 0
28
2 Treated 276.67 222.00 80.24 49 56.6
days
1 After Control 739.33 631 85.35 0 0
42
2 Treated 247.33 203 82.08 66.5 67.8
days

90 Fig. 2. Regression graph showing toxicity

80 f(x) = 1149.41652349951 x + 14.2330705552386 of cypermethrin against T. confusum after
R² = 0.96795078700794
70
48 hours.
60
50
%Mortality

100
40
90 f(x) = 1186.56412396483 x + 20.5560345824885
30 R² = 0.947145465105372
80
20 70
10 60
%Mortality

0 50
0 0.01 0.02 0.03 0.04 0.05 0.06 0.07
40
% Concentration 30
20
10
0
Fig. 1. Regression graph showing toxicity 0 0.01 0.02 0.03 0.04 0.05 0.06 0.07

of cypermethrin against T. confusum after % Concentration

24 hours.

Fig. 3. Regression graph showing toxicity

of cypermethrin against T. confusum after
100
90
72 hours.
f(x) = 1136.3376403961 x + 18.6883306550064
80 R² = 0.951881917003204
100
70
90
%Mortality

60 80
50 70
40 60
30
ortality

50
20 40
%M

f(x) = 3965.75660390336 x − 30.6912221532478

10 30 R² = 0.931335628438842
0 20
0 0.01 0.02 0.03 0.04 0.05 0.06 0.07
10
% Concentration 0
0.80% 1.00% 1.20% 1.40% 1.60% 1.80%

% Concentration

14
Fig. 4. Regression graph showing toxicity 100
80
of L. camara against T. confusum after 24 60

Mortality %
hours. 40
20
0 Day 1
Day 2
100 Day 3
90 Day 4
80 Day 5
70
60
f(x) = 4310.64211324934 x − 18.2767777088098
%Mortality

50 R² = 0.935628742514984
40 Concentration %
30
20 Fig. 7. Cypermethrin residual effects
10
0
against T. confusum
0.80%1.00%1.20%1.40%1.60%1.80%2.00%
% Concentration
60
Fig. 5. Regression graph showing toxicity
of L. camara against T. confusum after 48 50
Mortality %

hours.
40
Day 1
100
30 Day 2
90
Day 3
80 20
70 Day 4
60 f(x) = 4464.44515875569 x − 18.2077931354694 10 Day 5
%Mortality

50 R² = 0.958588957055214
0
40 1.00% 1.20% 1.40% 1.60% 1.80%
30
Concentration %
20
10
Fig. 8. L. camara residual effects against
0
T. confusum
% Concentration
1 HOUR AFTER 5 HOUR AFTER
80.00
Fig. 6. Regression graph showing toxicity
70.00
of L. camara against T. confusum after 72
60.00
hours.
50.00
40.00
30.00
20.00
10.00
0.00
1 2 3 4 5

15
Fig. 9. Cypermethrin repellency against T. CONTROL TREATED
confusum after 1 and 5 hours. 100
80

Fecundity
60
1 HOUR AFTER 5 HOUR AFTER 40
70.00 20
60.00 0
14 28 42
50.00
40.00
Days
30.00
20.00 Fig. 11. Fecundity effects of cypermethrin
10.00 extract against T. confusum after 14, 28
and 42 days of LC50 dose treatment by
0.00
1% 1% 1% 2% 2% filter paper technique.

Fig. 10. L. camara repellency against T. CONTROL TREATED

confusum after 1 and 5 hours. 100
80
Fecundity

60
40
20
0
14 28 42
Days

Fig. 12. Fecundity effects of L. camara

extract against T. confusum after 14, 28
and 42 days of LC50 dose treatment by
filter paper technique.

Discussion

Toxicity with increased concentration rate, which is reported in previous researches.

LC50 value for cypermethrin was calculated to be 6.1 µg/cm2 after 24 hrs, 9.6 µg/cm2 for
imidacloprid, 298.8 µg/cm2 for L camara and 342 µg/cm2 for A. javanica after 48 hrs. The
present investigation is similar to the previous study. However, test insect and test
compounds were different. LC50 to be 6.1µg/cm2, while neem extract caused 34% mortality
on the tested insect. In our study LC 50 value for cypermethrin was calculated to be 6.1µg/cm2
after 24 hrs, while L. camara extract caused 64.2% mortality against T. confusum. The

16
present study is in agreement with the previous investigation that pyrethroids are most toxic
than a plant extract. Five concentrations ranged from 1-10% w/w were prepared compared
with untreated and an insecticide actellic super for positive control of five-eight days old
insects used in the experiment. Percent mortality caused by L. camara and T. vogelii was
90% and 93 %, respectively, on 21 days of exposure (Kachhwaha et al., 2015). LT50 for both
extracts was five-six days at a concentration of 7.5-10%.and seven-eight days at a
concentration of 2.5-5 w/w. Their results showed that a good relationship between mortality
and concentration was found. In the present work, L. camara produced 64.2% mortality at the
highest dose of 358.5µg/cm2 after 72 hrs against T. confusum. The results of the current study
are not similar to the previous investigation due to different test insect and different solvents
used; however, a dose-response relationship was found, which an agreement with the
previous work is. Athanassiou et al. (2004) evaluated the residual toxicity of pyrethroids i-e
alpha-cypermethrin, beta-cyfluthrin and deltamethrin against T. confusum. Cypermethrin
produced mortalities of 86.2%, 89.66%, 48.28%, 31.04% and 10.35% after, 1, 2, 3, 4 and 5
days of treatment at the highest dose of 12.5µg/cm 2, respectively. Islam and Talukder (2005)
compared the residual toxicity of Azadirachta indica, Cynodon dactylon and Tagetes erecta
with Malathion and Carbaryl against T. castaneum. In the present study, residual toxicity of
L. camara and A. javanica was tested against T. confusum. Maximum residual toxicity for L.
camara was observed 48.28% and A. javanica 43.33% at 1.8% concentration after 48 hrs and
72 hrs, respectively. Khanam et al. (2008) evaluated the repellent activities of rhizome and
leaves extract of Zingiber cassumunar in different solvents on T. castaneum and T. confusum
showed repellency to the rhizome and leaves extract of Z. cassumnar. Ogendo et al. (2003)
reported the fecundity effects of methanol extracts of two plants Tephrosia vogelii and
Lantana camara, against maize weevil Sitophilus zeamais reduced F1 progeny up to 75 % as
compared to untreated control. Maji et al. (2014) investigated the ethanol extracts of 06
plants i-e Lantana camara, eucalyptus leaves, turmeric, garlic, and neem, against biological
parameters of Callosobrucus chinensis where no adult formation (0.00%) was recorded while
4% adult emergence for neem (4.76%) and turmeric (6.67%) were noted (Kachhwaha et al.
2015).

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