Sarhad J. Agric. Vol.28, No.

2, 2012

EFFECT OF EGG PARASITOID, TRICHOGRAMMA CHILONIS, IN COMBINATION

WITH CHRYSOPERLA CARNEA AND NEEM SEED EXTRACT AGAINST TOMATO
FRUITWORM, HELICOVERPA ARMIGERA
MUHAMMAD USMAN, MIAN INAYATULLAH, AMJAD USMAN
KAMRAN SOHAIL and SYED FAHAD SHAH
Department of Entomology, Agricultural University, Peshawar – Pakistan.
E-Mail: sohailagrian@gmail.com
ABSTRACT
To investigate the efficiency of Trichogramma chilonis, T. chilonis in combination with Chrysoperla
carnea and neem extract against tomato fruitworm, Helicoverpa armigera, field experiments were carried out at
the Research Farm of Agricultural University, Peshawar, Pakistan during summer 2009. The plot size was 8 x 5
meters. Five treatments were used i.e. trichocard having 300 parasitized eggs (T1), trichocard having 300
parasitized eggs in combination with 45 Chrysoperla 2nd instar larvae (T2), trichocard having 300 parasitized
eggs and neem extract (T3), trichocard having 300 parasitized eggs in combination with 45 Chrysoperla larvae,
neem extract (T4) and control (T0). The lowest number of H. armigera larvae (0.68 plant-1) recorded in T4 was
significantly lower than those of all other plots which were followed by T3 and T2 i.e. 0.85 and 0.97 plant-1 while
the highest number of larvae plant-1 (1.40) was recorded in T0. The yield of tomato was highest (11570 kg)
recorded in T4 and the lowest yield (7250 kg) was recorded in T5. The percent weight loss was minimum
(11.04%) in T4 while maximum (20.45%) in T0. Based on yield and percent fruit damage, treatment having
trichocard having 300 parasitized eggs in combination with Chrysoperla and neem extract is the most
promising for effective management of H. armigera on tomato.
Key Words: Egg parasitoid, Trichogramma chilonis, Chrysoperla carnea, neem, Helicoverpa armigera.
Citation: Usman, M., M. Inayatullah, A. Usman, K. Sohail and S.F. Shah. 2012. Effect of egg parasitoid,
Trichogramma chilonis, in combination with Chrysoperla carnea and neem seed extract against tomato
fruitworm, Helicoverpa armigera. Sarhad J. Agric. 28(2):253-257
INTRODUCTION
Tomato, Lycopersicum escolentum Mill, is an important and widely used vegetable crop. It is very
nutritive and delicious; very few vegetables can match its nutritional value. Tomato is one of the most important
vegetable crops cultivated for its fleshy fruits and considered as important commercial and dietary vegetable
crop. It is short duration crop and gives high yield, it is important from economic point of view and hence area
under its cultivation is increasing day by day (Kevany et al., 2008).
In Pakistan the fruit borer Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is a key pest of
several crops. Tomato, cotton, pigeon pea, and chickpea are consistently suffered by this insect in different parts
of the country. The fruit borer is destructive in larval stages and cause serious damage to tomato fruit. The moth
(adult) generally appears during April-May and lay eggs on leaves. The young larvae feed on leaves, flowers
and bore in tomato fruit and complete most of its development inside the fruit. Larvae have 5-6 instars, all larval
stages damage tomato fruit. During development the larvae may emerge from one fruit and enter another and
cause infestation. The last larval instars come out from the tomato fruit, falls down to the soil and pupate in soil.
A single caterpillar can damage up to 12 fruits and hence lowers the market value of the fruit and ultimately
farmers bear great losses (Hussain and Bilal, 2007).
Among the parasitoids the Trichogramma species are the most commonly used egg parasitoids in
biological control program released on a large scale against lepidopterous pests. Adults of T. chilonis
(Hymenoptera: Trichogrammatidae) are tiny wasps that kill the eggs of hosts by laying one or more of their own
eggs inside the much larger eggs of Lepidoptera. Recognizing the potential of the Trichogramma species as a
biological control agents, entomologist in the early 1900 began to mass rear Trichogramma for insect control.
Today Trichogramma species are the most widely used insect natural enemy in the world because their mass
rearing is easy on one hand and they attack many important crop insect pests on other hand (Ayvaz et al., 2008).
Chrysoperla or green lacewing belongs to the insect order Neuroptera and is found in most
environments throughout the world. There are several species and the most common is C. carnea Stephen. The
adults feed on pollen, nectar or honeydew of aphids, but the larvae are voracious predators (Easterbrook et al.,
2006). Neem, a plant originated (botanical) pesticides which is environment friendly and is well known for its
diverse pest control properties. It works as an insect feeding deterrent, but in various forms it also serves as a
repellent, growth regulator, oviposition (egg deposition) suppressant, sterilant, or toxin. Neem caused higher
reduction in the larvae of shoot and fruit borer Earias vittella and E. insulana and recorded the highest yields of
 M. Usman et al. effect of egg parasitoid, trichogramma chilonis against tomato fruitworm… 254

4600 and 4180 kg ha-1 in okra crop (Dhingra et al., 2008).

In Pakistan tomato pests are usually controlled by using chemicals and no serious efforts have been
made to use non chemical methods. Published information shows that efforts have been made in many countries
of the world to control tomato fruit worm by using natural enemies including C. carnea and Trichogramma. In
Khyber Pakhtunkhwa Inayatullah (2007) studied the effect of Trichogramma alone and in combination with C.
carnea on the tomato fruit worm and reported significant reduction in fruit worm infestation and percent yield
loss. The author, however did not record any significant increase in yield. In the present studies the effect of
Trichogramma alone and the combined effect of Trichogramma with Chrysoperla and neem seed extract were
studied. In the present research, we tried:
i. To study the effect of T. chilonis alone and in combination with C. carnea, and neem extract against tomato
fruit worm, H. armigera, at the Research Farm of the Khyber Pakhtunkhwa Agricultural University
Peshawar, and
ii. to study the effect of T. chilonis alone and in combination with C. carnea, and Neem extract on the yield of
tomato.
MATERIALS AND METHODS
Experiments were conducted at the Research Farm of Agricultural University, Peshawar, Pakistan during
summer 2009. The experiment was composed of 5 treatments each with 3 replications. The plot size was 8 x 5
m and between each treatment a 3 meters buffer zone was maintained. The distances between the replications
were 2 meters. The experiment was designed as RCB Design.
Rearing of Trichogramma and C. carnea
The rearing of both Trichogramma and Chrysoperla was done at the research laboratory of the
Department of Entomology, Agricultural University, Peshawar, Pakistan on standard diet by the procedure
adopted by Inayatullah, (2007).
Field Preparation and Release of Trichogramma and Chrysoperla
The field was prepared for tomato with normal cultural practices. The experiment was conducted on well-
prepared seedbed, ploughed twice for obtaining bumper crop. Transplantation was done in the month of March.
The release of trichocards was started when the tomato crop reached the flowering stage. Total six releases were
done at ten days interval throughout cropping season. The trichocards (from the laboratory culture) from which
the emergence was expected in few hours were selected in laboratory; the required numbers of parasitized eggs
on card were counted. Trichocard having number of 300 parasitized eggs, in combination with Chrysoperla and
neem extract were applied to each plot except control. The trichocards were hooked up with the metal rod and
were placed in the center of each plot. The lower portion of the rod was greased to prevent the access of ants and
any other crawling insects. The required numbers of second instars larvae of C. carnea were obtained from the
Chrysoperla culture established in the entomology research laboratory and were taken to the field. Test tubes
containing the second instar larvae were shaken on the leaves of plants at different sites in the same plot.
Preparation of Neem Extract
Mixture of half Kg crushed neem seed and 10g surf (detergent) were tightened in muslin cloth and placed
in 5 liter water for 24h. This gave us 10% solution of neem extract using the following formula (Shah, 2004).
V1C1=V2C2
V1= volume of water which is required
V2= volume of known quantity
C1= given concentration
C2= required concentration
Data Collection
Ten plants plot-1 were selected randomly and the number of larvae on all the selected plants were counted
and recorded at one week interval. This process was repeated for all the six releases. The mature tomato fruit
were collected separately from each plot. Weight and number of damaged tomato fruit were recorded for each
plot separately. The total yield of tomato was determined by adding the yields of tomato per picking of each
plot.
Yield = Total weight of tomato collected from each plot
Percent weight loss = wt. of damage fruit x 100
Total wt of tomato
Sarhad J. Agric. Vol.28, No.2, 2012 255

Treatments
T1 Trichocard having 300 parasitized eggs
T2 Trichocard having 300 parasitized eggs in combination with 45 Chrysoperla 2nd instar larvae
T3 Trichocard having 300 parasitized eggs in combination with Neem extract
T4 Trichocard having 300 parasitized eggs in combination with 45 Chrysoperla 2nd instar larvae and neem
extract
T0 Control (no trichocards)
RESULTS AND DISCUSSION
Data collected on 1st week after the treatment indicated that the lowest number of H. armigera larvae
-1
plant recorded in treatments T3 and T4 i.e. recorded 0.50 and 0.60. The treatments T1 and T2 on par with T0
control which recorded 0.90 larvae plant-1. The treatments T3 and T4 were significantly different from the T0
Table I.
Table I Field layout of the experiment
T1 T2 T3 T4 T0
300 Trichogramma
300 Trichogramma
eggs + 45
300 Trichogramma eggs + 45 300 Trichogramma
Rep 1 Chrysoperla 2nd Control
eggs Chrysoperla 2nd eggs + neem extract
instars larvae and
instars larvae
neem extract
300 Trichogramma
300 Trichogramma
eggs + 45
300 Trichogramma 300 Trichogramma eggs + 45
Rep 2 Chrysoperla 2nd Control
eggs + neem extract eggs Chrysoperla 2nd
instars larvae and
instars larvae
neem extract
300 Trichogramma
300 Trichogramma
eggs + 45
eggs + 45 300 Trichogramma 300 Trichogramma
Rep 3 Control Chrysoperla 2nd
Chrysoperla 2nd eggs eggs + neem extract
instars larvae and
instars larvae
neem extract
Data in Table II recorded on 2nd week revealed the lowest number (0.63) of H. armigera larvae plant-1
was recorded in T4 followed by T3 i.e. 0.83 larvae plant-1. The number of H. armigera larvae plant-1 recorded on
T0 (0.93 larvae plant-1) was at par with T1 and T2. Significant difference was recorded between the T3, T4 and T0.
The number of H. armigera larvae plant-1 recorded on 3rd week was highest in T1 i.e. recorded 1.03
larvae plant-1 followed by 0.97 and 0.87 at T2 and T3 whereas the lowest number of 0.73 larvae plant-1 was
recorded in T4. The number of H. armigera larvae plant-1 recorded in T0 was 1.40. All the treatments were
significantly different with the T0.
Table II Effect of Bio agents and neem extract on the infestation of tomato fruit worm, Helicoverpa armigera,
plant-1 recorded on different dates during 2009
Treatments 15th April 22nd April 29th April 5th
May 12th May 19th May 26th May 3rd June 10th June Mean
T1 0.90a 1.03a 1.20b 1.33b 1.50b 1.37b 1.13b 0.97b 0.67b 1.12b
T2 0.80a 0.93a 0.97c 1.23b 1.40b 1.20b 0.90bc 0.70c 0.67b 0.97c
T3 0.60b 0.83ab 0.87cd 0.93c 1.10c 0.97c 0.90bc 0.77c 0.67b 0.85d
T4 0.50b 0.63b 0.73d 0.67d 0.90d 0.77d 0.80c 0.67c 0.50b 0.68e
T0 0.90a 0.93a 1.40a 1.63a 1.83a 1.67a 1.67a 1.57a 1.00a 1.40a
Means followed by the same letters within each column are not significantly different at 5 % level of probability (LSD –Test).
Results in Table II on 4th week indicated that the lowest number of H. armigera larvae was recorded in
T4 i.e. 0.67 larvae plant-1 which was followed by 0.93 larvae plant-1 at T3. The number of larvae recorded on T1
and T2 were 1.33 and 1.23 larvae plant-1 as compared to the T0 where the number of larvae recorded 1.63 larvae
plant-1. Statistical analysis of the data showed significant difference between the treatments and T0. Data
recorded on 5th week after the application showed the highest (1.50) number of H. armigera larvae plant-1 in T1
followed by 1.40 and 1.10 larvae plant-1 in T2 and T3 as compared to T4 where the number of H. armigera larvae
was recorded lowest (0.90 larvae plant-1). In T0 the number of H. armigera larvae was 1.83 larvae plant-1. Data
collected on 6th week revealed that the lowest number of H. armigera larvae plant-1 was recorded in T4 i.e. 0.77
larvae plant-1 followed by 0.97 larvae plant-1 at T3. The number of larvae recorded in T1 and T2 and T0 were 1.37,
1.20 and 1.67, respectively. The number of H. armigera larvae on 7th week was highest (1.13 larvae plant-1) in
T1 as compared to T0 where the number of larvae recorded was 1.67 larvae plant-1. The number of larvae plant-1
0.90 recorded in T2 on par with T3 and T4. On 9th week the highest number of larvae plant-1 was recorded in
control i.e. 1.57 larvae plant-1. The number of larvae 0.70 plant-1 recorded in T2 on par with T3 and T4. On 10th
week the number of larvae 0.67 larvae plant-1 recorded in T1 was also on par with T2, T3 and T4. In T0 the
M. Usman et al. effect of egg parasitoid, trichogramma chilonis against tomato fruitworm… 256

number of larvae recorded was 1.00 larvae plant-1. In both weeks all the treatments were significantly different
from the control.
Results concluded from overall mean of the data indicated that the lowest number of H. armigera
larvae was recorded in T1 i.e. recorded 1.12 larvae plant-1 followed by 0.97 and 0.85 larvae plant-1 in T2 and T3 as
compared to T4 where the number of larvae was recorded lowest (0.68 larvae plant-1). In control the number of
larvae recorded was 1.40 larvae plant-1. The treatments T1, T2, T3 and T4 were significantly different from the
control.
Yield and Percent Weight Loss
The mean yield of tomato fruit recorded in T1, T2, T3 and T4 were 8428, 9039, 10010 and 11570 kg
respectively, as compared to control plot where the yield of tomato fruit was 7250 kg. Table II shows that
highest yield (11570 kg ha-1) was recorded in T4 which was non-significant from the yield recorded in T3 (10010
kg/ha) while significantly higher than T1, T2 and T0. Percent weight loss of tomato fruit recorded in T1, T2, T3
and T4 were 15.18, 13.65, 12.59 and 11.04%, respectively as compared to T0 plot where the percent weight loss
of tomato fruit was 20.45%. Statistical analysis of the data showed that the lowest percent damage (11.04%)
recorded in T4 was non-significant from T3 (12.59%) and significant with all other treatments.
Table III Effect of Bio agents and neem extract on tomato yield and percent weight loss by H. armigera during 2009
Treatments Yield (Kg) % weight loss
T1 107000 Trichogramma parasitized eggs/ hectare 8428 BC 15.18 B
T2 107000 Trichogramma parasitized eggs with 66000/ha Chrysoperla 9039 BC 13.65 BC
T3 107000 Trichogramma parasitized eggs with neem extract 10010 AB 12.59 CD
T4 107000 Trichogramma parasitized eggs with 66000/ha Chrysoperla, neem extract 11570 A 11.04 D
T0 Control 7250 C 20.45 A
Means followed by the same letters are not significantly different at 5 % level of probability (LSD –Test).
The above results show that Trichogramma parasitism, Trichogramma in combination with
Chrysoperla and neem (T4) does have an impact on population of tomato fruit worm. In the present experiment
Trichogramma in combination with Chrysoperla and neem extract (T4) decreases the population of tomato fruit
worm. These results are in conformity with those of Reddy and Manjunatha (2000) who used Trichogramma
chilonis alone and in combination with plant extracts and synthetic pesticides and reported positive results in
controlling lepidopterous pests of field crops. Shah (2008) after conducting field experiments also reported that
Chrysoperla carnea and Trichogramma chilonis are the best bio-control agents for controlling tomato fruit
worm in tomato crop. For significant reduction in pest population however, application of 50 2nd instar C.
carnea larvae and 400 Trichogramma wasps were significant in reducing the pest population. Despite
application of bio-control agent the pest survived, though in significantly less number, to cause some damage.
Workers around the world could not achieve 100% control with Trichogramma, Chrysoperla and neem extract.
This includes Shah (2008) who tested Trichograma, Chryosperla and neem extract respectively, against H.
armigera.
In the present studies the mean number of tomato fruit worm recorded show that the application of
Trichogramma in combination with Chrysoperla and neem (T4) significantly help in reducing the infestation of
tomato fruit worm. Although the treatment with the Trichogramma in combination with Chrysoperla and neem
(T4) significantly reduced the number of tomato fruit worm, it also put a positive impact on yield. Shah (2008)
achieved significant increase in yield.
In the present experiment, the reason for this low-significant reduction may be that due to less number
of pest larvae, they faced little competition and distributed well in the field plant-1 and caused damage. Figures
of percent weight loss show that all the treatments are better than control and application of Trichogramma in
combination with Chrysoperla and neem can be recommended. Release of Trichogramma in combination with
Chrysoperla and neem put a positive impact on reducing fruit worm infestation, yield and percent weight losses.
CONCLUSION AND RECOMMENDATIONS
Trichogramma and C. carnea in combination with neem seed extract work well against tomato fruit
worm and can be incorporated into and IPM strategy directed towards managing tomato fruit worm and can also
be used in the biological control programme. Despite the application of these bio-control agents losses due to
fruit worm were still high. More work should be done to enhance the efficacy of these bio-control agents.
Cannibalism exists in C. carnea, it is impossible to rear two or more C. carnea in a test tube which is extremely
laborious, costly and time consuming. Work should be done to minimize or modify this cannibalism behavior of
C. carnea.
Sarhad J. Agric. Vol.28, No.2, 2012 257

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