Koya University

Faculty of Science and Health

Biology Department

Title
Controlling of Thaumetopoea Solitaria With Crude Plant Extract

A research project or project submitted to the department of Biology, Faculty of

Science and Health, Koya University in partial fulfilment of the requirements for
the degree Bachelor in Science - Biology

Prepared by
Muhamad Ibrahim Malla

Supervised by

Mr. Muzafar Kanabi

2021-2022
 Acknowledgements

First of all, thanks to Allah who gives me courage to done this project then I want to thanks and
dedicate this project to people who helped me to continue this project.

To my dear mother, who still beacon to me in life and to my dear father, may Allah prolong their
life. This research gave to them a symbol of love and fulfilment and recognition of aboriginal me
on side and to you gave me all the love and all the support.

To my dear supervisor Mr. Muzafar Kanabi Omer without his support and work this project will
not be completed.

And to all my friends who helped me and constantly completion and continuation of this
scientific humble effort.

ii
 Table of Contents

ACKNOWLEDGEMENTS ................................................................................................ II

ABSTRACT ......................................................................................................................... V

1. INTRODUCTION ............................................................................................................ 1

2.MATERIAL AND METHODS ........................................................................................ 3

2.1 Plant Extract .............................................................................................................................. 3

2.2 Insect Collection ........................................................................................................................ 4

2.3 Mortality Test ............................................................................................................................ 5

3.RESULTS AND DISCUSSION ........................................................................................ 6

4.CONCLUSION ................................................................................................................ 13
4.1 Conclusion .............................................................................................................................. 13

5. REFERENCES ............................................................................................................... 14
5.1 References .............................................................................................................................. 14

iii
 List of Figures
Figure No. Figure’s Title Page No.
2-1 Plant Extract 3
2-2 Insect Collection 4
2-3 Mortality Test 5

List of Figures

Figure No. Figure’s Title Page No.

2-1 Plant Extract 3
2-2 Insect Collection 4
2-3 Mortality Test 5

3.1 1stage flower 6

3.2 1stage immature leaf 7
3.3 1stage mature leaf 7
3.4 1stage immature fruit 8
3.5 1stage mature fruit 8
3.6 2stage flower 9
3.7 2stage immature leaf 10
3.8 2stage mature leaf 10
3.9 2stage immature fruit 11
3.10 2stage mature fruit 11

iv
Abstract

Thaumetopoea solitaria is a severe pest in the United States. Pistachios can be found all
across the Mediterranean and its environs. The purpose of this study was to see how melia
azadarach affected thaumetopoea solitarea larvae as an alternative control approach with
minimal unfavorable side effects. Two larvae stages were tested with various concentration
of this plant extract and different part of this plant under controlling condition by rushing
them inside petri dish. The effect of melia azadarach was significantly higher on first stage
larvae then second stage. High larval mortality was achieved in first 24 hours after
treatment especially for the first larval stage with the application of the highest 3
concentration (25%, 50%, 100%). According to the findings Melia azadarach is a strong
option for decreasing T.solitaria populations in pistachio orchards, and could be employed
as a biological control agent.

v
 1. Introduction

Iran, Turkey, USA and Israel are the largest producers of pistachio which is important both
as a commodity for producer countries and in national and international trade markets [
FOA (2006)]. The most common insect pest of pistachio in Iran [Davatchi AG (1958) ] ,
Turkey[ Mart C, çelık MY, yığıt A (1995): Mart C, Karaat (1990)], as well as Greece
[Maurikies PA, Tsourgianni A(1998) ] and Israel [Haperin J(1983) ] are Thaumetopoea
solitaria (Lepidoptera : Thaumetopoeidae). This insects larvae damage many parts of
pistachio like buds and leaves until they grow, resulting in a reduced number of shoots and
stagnation in tree development. Overall the scientists discovered that T.solitaria reduce the
yield of pistachio nuts [Mart C, Eskalen A (2000): Uygun N, Ulusoy MR, karaca I (2002)].
Over the last 50 years prior insects pest have been examined with synthetic insecticides to
discovery of organochlorin and organophosphate insecticide in the late 1930 and early
1940s . botanical insecticide are significant product for pest management in industrial
countries [Isman MB(2006)]. Most insecticide compound are related to 4 main classes :
organochlorines, organophosphates, carbamats , and pyrethroids , among this major classes
that use today are organophosphate and carbamates [Ware GW(1998); Dorow E(1993)]. In
pesticide management there are many negative effect on non-target organisms that could be
present including human and environment [Rembold H (1984): FAO(1992)].The organic
synthetic insecticide are more hazardous to handle leave toxic residues in food product and
are not easily biodegradable, in addition to their deleterious influence on the environment,
unlike synthetic ones that kill both pests and predators, natural insecticides are relatively
enactive against the litter. The botanical insecticides are generally pest-specific and are
relatively harmless to non-target organisms include human, they are also biodegradable and
harmless to the environment [Jacobson M(2004)]. Furthermore, unlike conventional
insecticide that are based on a single active ingredient, plant-derived insecticide comprise
an array of chemical compound which act concertedly on both behavioral and physiological
processes, thus the chances of pest developing resistance to such substances are less likely
[Saxena RC(1987)]. Same plant have substances with wide range of activities; for example
extract from the neem tree Azadirachta indica are antifeedant , antioviposition, repellent
and grouwth-regulation [Rembold H(1984); Franzen H(1993)]. In contrast, the toxicity of
conventional synthetic insecticide is mainly restricted to neuro-muscular function [Ware
GW(1998)]. Conventional synthetic insecticide require special safety procedures and
1
equipment during production and application because of exposure risks for human and
environment [Franzen H(1993)] and food [FAO(1992)]. The synthetic insecticides are
expensive and have in many cases only produced moderate results along with major
ecological damage [Franzen H(1993)]. In contrast, the low toxicity of insecticides makes
the processing and botanical application of the product inexpensive. In many cases, the
materials are locally available and affordable [Childs FJ. Chamberlain JR(2001)]. The
supply of natural insecticides could be made continuous at a cheaper rate by regular
cultivation. The use of organochlorine insecticides has been banned in developed
countries, and alternative methods of insect pest control are being investigated [Franzen
H(1993)]. Botanicals are a promising source of pest control compounds. The pool of plants
possessing insecticidal substances is enormous [FAO(1992)]. These have generated
extraordinary interest in recent years as potential sources of natural insect control agents.
Today over 2000 species of plants are known to possess some insecticidal activity (15). The
main purpose of agricultural studies is to increase the yield of product.

2
 2.Material and Methods

2.1 Plant Extract

Firstly, fresh plant parts of (Melia azedarach) such as pre-mature leaf, mature leaf, flower,
pre-mature fruit, and mature fruit were collected from Koya town. Then, plant parts washed
with water, dark-shade dried, and ground to powder by using an electric blender. After that,
about 20gm of each plant part’s powder was soaked in 100ml of tap water in different glass
bottles. After one day the solutions were filtered by using filter paper and stored in the glass
container in a refrigerator (4co) (Negahban M., Moharramipour S., Sefidkon F.’ 2007).
Plant part extracts were tested to determine the mortality percentage of Thaumetopoea
solitarea larvae.

Figure 1

3
2.2 Insect Collection
Two to three weeks larvae of (Taumetopoea solitarea) were collected and separated from
the infected pistachio tree [Madkour HM, Zaitoun AA and Singer F.( 2013)]. The insect
was reared at a hot place approximately between (20-25)℃ prior to the test.

Figure 2

4
2.3 Mortality Test
Plant part extracts were used to test the larvae mortality rate of (Taumetopoea solitarea).
The extracts were changed to different concentration (0%, 25%, 50% and 100%).Then
prepare 60 petri dish it means for each concentration and plant part three time repeat it for
more accurate our result and filter paper placed inside the each petri dish (6 cm diameter)
and then 5 larvae of the insect were released into each petri dish after that they were
sprayed with each concentration from each part of the plant until the insect body was
rushed. The same number of insects were treated with tap water and used as a control in the
test. Four replications were done for each concentration and control in each part of the plant
(Sahaf, et al, 2008). The larvae mortality of insects was recorded at each hour in total 24
hours after treatment. The number of insect mortality was measured by means of direct
observation, and when the insect has no motion, the insect was dead [Muhammad, (2008)].

Figure 3

5
 3.Results and discussion

Statistical analysis showed that there was a significant difference in larvae mortality
between the concentration of each plant part on both stages. We are illustrating the results
in each 6 hour after treatment for totally 24 hours, we repeated each test three times for
each concentration which contain 5 larvae, the average death rate for total 15 larvae as
follows, in the first stage :

Flower part

For (0%) concentration in the first 6H are 0, in 12H are 1, in 18H are 4, in 24H are 4.
For (25%) concentration in 6H are 2, in 12H are 5, in 18H are 5, in 24H are 6.
For (50%) concentration in 6H are 2, in 12H are 4, in 18H are 6, in 24H are 9.
For (100%) concentration in 6H are 6, in 12H are 8, in 18H are 9, in 24H are 11.

1.2

0.8 control

0.6 25%
50%
0.4
100%
0.2

0
1hr 6hr 12hr 24hr

Figure 4

6
Immature leaf

For (0%) concentration in 6H are 0, in 12H are 0, in 18H are 2, in 24H are 3.
For (25%) concentration in 6H are 2, in 12H are 3, in 18H are 5, in 24H are 5.
For (50%) concentration in 6H are 4, in 12H are 4, in 18H are 7, in 24H are 8.
For (100) concentration in 6H are 5, in 12H are 7, in 18H are 8, in 24H are 8.

1.2

0.8 control

0.6 25%
50%
0.4
100%
0.2

0
1hr 6hr 12hr 24hr

Figure 5

Mature leaf

For (0%) concentration in 6H are 0, in 12H are 0, in 18H are 2, in 24H are 2.
For (25%) concentration in 6H are 1, in 12H are 1, in 18H are 1, in 24H are 1.
For (50%) concentration in 6H are 6, in 12H are 10, in 18H are 12, in 24H are 12.
For (100%) concentration in 6H are 4, in 12H are 6, in 18H are 10, in 24H are 12.

1.2

0.8 control

0.6 25%
50%
0.4
100%
0.2

0
1hr 6hr 12hr 24hr

Figure 6
7
Immature fruit

For (0%) concentration in 6H are 0, in 12H are 1, in 18H are 2, in 24h are 2.
For (25%) concentration in 6H are 2, in 12H are 3, in 18H are 3, in 24H are 4.
For (50%) concentration in 6H are 4, in 12H are 5, in 18H are 5, in 24H are 6.
For (100%) concentration in 6H are 9, in 12H are 11, in 18H are 12, in 24H are 12.

1.2

0.8 control

0.6 25%
50%
0.4
100%
0.2

0
1hr 6hr 12hr 24hr

Figure 7

Mature fruit

For (0%) concentration in 6H are 2, in 12H are 2, in 18H are 2, in 24h are 2.
For (25%) concentration in 6H are 0, in 12H are 2, in 18H are 6, in 24H are 7.
For (50%) concentration in 6H are 7, in 12H are 11, in 18H are 11, in 24H are 13.
For (100%) concentration in 6H are 5, in 12H are 10, in 18H are 10, in 24H are 10.

1.2

0.8 control

0.6 25%
50%
0.4
100%
0.2

0
1hr 6hr 12hr 24hr

Figure 8

8
The data indicate that noticeable mortality for the first stage are started in the first 6 hours
after rushing larvae of T.solitaria of all plant parts especially in the concentration of 50 and
100 it was more evident then the other concentration due to large number of death rate
which are quite enough for knowing that the plant extraction was worked very good on this
stage and the death rate rise during 12,18,24 hours. In other concentration also there was a
good result for the morality of larvae but they are noticed more in 18 hours after rushing.
However , the highest concentration showing more death rate. Over all according to this
information the greatest number of larvae are killed by mature fruit extraction section at
this stage it implies that have a significant impact then others

For the second stage this data are recorded for death of larvae

Flower part

For (0%) concentration in the first 6H are 0, in 12H are 0, in 18H are 0, in 24H are 0.
For (25%) concentration in 6H are 3, in 12H are 3, in 18H are 4, in 24H are 5.
For (50%) concentration in 6H are 0, in 12H are 1, in 18H are 2, in 24H are 3.
For (100%) concentration in 6H are 0, in 12H are 1, in 18H are 2, in 24H are 2.

1.2

0.8 control

0.6 25%
50%
0.4
100%
0.2

0
1hr 6hr 12hr 24hr

Figure 9

9
Immature leaf

For (0%) concentration in 6H are 0, in 12H are 0, in 18H are 0, in 24H are 1.
For (25%) concentration in 6H are 0, in 12H are 0, in 18H are 0, in 24H are 3.
For (50%) concentration in 6H are 0, in 12H are 0, in 18H are 0, in 24H are 0.
For (100) concentration in 6H are 0, in 12H are 0, in 18H are 0, in 24H are 1.

1.2

0.8 control

0.6 25%
50%
0.4
100%
0.2

0
1hr 6hr 12hr 24hr

Figure 10

Mature leaf

For (0%) concentration in 6H are 0, in 12H are 0, in 18H are 0, in 24H are 1.
For (25%) concentration in 6H are 0, in 12H are 1, in 18H are 1, in 24H are 6.
For (50%) concentration in 6H are 0, in 12H are 2, in 18H are 2, in 24H are 6.
For (100%) concentration in 6H are 0, in 12H are 0, in 18H are 0, in 24H are 0.

1.2

0.8 control

0.6 25%
50%
0.4
100%
0.2

0
1hr 6hr 12hr 24hr

Figure 11
10
Immature fruit

For (0%) concentration in 6H are 0, in 12H are 0, in 18H are 0, in 24h are 0.
For (25%) concentration in 6H are 1, in 12H are 2, in 18H are 4, in 24H are 5.
For (50%) concentration in 6H are 0, in 12H are 2, in 18H are 3, in 24H are 3.
For (100%) concentration in 6H are 0, in 12H are 5, in 18H are 5, in 24H are 6.

1.2

0.8 control

0.6 25%
50%
0.4
100%
0.2

0
1hr 6hr 12hr 24hr

Figure 12

Mature fruit

For (0%) concentration in 6H are 0, in 12H are 0, in 18H are 0, in 24h are 3.
For (25%) concentration in 6H are 0, in 12H are 1, in 18H are 2, in 24H are 2.
For (50%) concentration in 6H are 0, in 12H are 0, in 18H are 0, in 24H are 0.
For (100%) concentration in 6H are 0, in 12H are 1, in 18H are 1, in 24H are 1.

1.2

0.8 control

0.6 25%
50%
0.4
100%
0.2

0
1hr 6hr 12hr 24hr

Figure 13
11
According to this data there was a very little quantity of death rate in every plant part and
every concentration during these stage. In all concentration there is only a little level of
mortality seen in the last 24 hours of flower, mature leaf and immature fruit only in contrast
with others which are very little or we can say they are zero. As a result we conclude that
Melia azadarach has no effect on this stage hence cannot be used to treat the problem of
T.solitaria .When [Jaqueline Scapinello, Jacir Dal Magro (2014)] in there study extract
supercritical carbon dioxide (SC-CO2) in melia azadarache as an insecticide agent against
fall armyworm (Spodoptera frugipedra). They indicated that mortality increase with the
increasing extract concentration with 50% mortality (LC50) at a concentration of
376,74mg/kg and reaching 100% mortality at 5000mg/kg. The inhibition of insect growth
was observed at higher concentration due to the antifeedant action of extract. Also
[Mohamed Farag, Mohamed H. Ahmad, Heba Yousef and A.-H. Abdel-Rahman (2011)]
was used oil of melia azadarach for killing (Spodoptera littoralis) he observed that they are
slightly very effective against 2 and 4 instar larvae in different concentration. This finding
is in agreement with results obtained by [Carpinella (2007)].

12
 4.Conclusion

4.1 Conclusion
 About 50% of larvae in the first stage are die, about16% of second stage are die.
 Increase concentration of plant extract increase death rate, also with passage of
more time after rushing death rate rises.
 Realy effect in first stage, it means they can be employed to support pests but only
in the early stage of larvae.

13
 5. References

5.1 References
1_ FOA (2006) food and agriculture organization of the united nations. FOA statistical
database, http://foastat.foa.org/

2_ Mart C, çelık MY, yığıt A (1995). biological observation and chemical control of
pistachio tig borer, kermania pistaciella Ams. injurious in pistachio orchards in turkey.
Acta Hortic 419:373-378

3_ Mart C, Karaat (1990) Guneydoğu Anadolu Bölgesi Antepfistıği alanlarinda

entomolojik sorunlar. Türkiye I. Antepfistığı Sim- pozyumu Bildirileri, Gaziantep, pp 160-
166

4_Maurikies PA, Tsourgianni A(1998) Chitzandis, A., piestechio nut insect pest and
mwans of control in Greece. Acta Hortiic 470:604-611

5_ Davatchi AG(1958) Etude Biologique de la faune Entomolojique des pistachio

sauvagest et Cultives Extrait de la Revue de pathologie vegetale et D'entomologie Agricole
de france, paris 37:49-52

6_ Haperin J(1983) Thaumetopoea solitaria freyer (Lepidoptera:Thaumetopoeidae) in

Israyl. phytoparasitica 11:71-82

7_ Mart C, Eskalen A (2000) Antepfıstığı zaralıları ve Hastalıkarı çifçi Broşürü. KSÜ

Bitki koruma Bölümü, Karamanmerş

8_ Uygun N, Ulusoy MR, karaca I (2002) Meyve ve Bağ zararlıları. ÇÜ ziraat fakültesi
Genel Yayin No :252 Adana

9_ Isman MB(2006), Neem and other botanical insecticides: barriers to commercialization,

Pyhtoparasitica 25:339-344, 1997

10_ Ware GW(1998), pesticides : Theory and application. Thompson publication, Fresno,
California: pp.308

14
11_Dorow E(1993). present practices of controlling desert locust outbreaks. In: New
strategies for locus control, Ed: Rembold H. ATSAF. Bonn: 89, pp 7-8

12_ Rembold H(1984). Secondary plant compounds on insect control with special reference
to azadirachtins. Advances in invertebrate reproduction 3:481-491

13_ Franzen H(1993). Need for development of new strategies for locust control. In: New
strategies for locust control. Ed: Rembold. H.ATSAF. Bonn;89. pp.9-13

14_ FAO(1992). Pesticide residues in food. Report no.116

15_ Jacobson M(2004) Insecticides from plant: A review of the literature. agricultural
handbook 461. US Department of Agriculture. Washington: 138 pp.

16_ Saxena RC(1987). Antifeedants in tropical pest management. Insect Sci Applic 8: 731-
736.

17_ Rembold H. Advances (1994) in invertebrate reproduction. Elsevier Science publishers

3: 481-491.

18_ schmutterrer H(1995). The neem tree Azadirachta indica A. Juss. and other meliaceous
plant. VCH Publishers: pp.696

19_ Childs FJ. Chamberlain JR(2001), Antwi EA et al. Improvement of neem and its
potential benefits to poor farmers. Department of International Development. UK: pp.32

20- Jaquelina scapinello , jacir dal dal margo(2014). the journal of supercritical fluids.
volume 93. page 20-26.

21- Mohamed Farag, Mohamed H. Ahmad, Heba Yousef and A.-H. Abdel-Rahman
(2011). zeitschrift fur naturforschung C, Tubungen. volum 129. page 406 4011.

22- Carpinella (2007). In vitro pediculicidal and ovicidal activity of an extract and oil from
fruit of Melia azadarach L. J. Am. Acad. Derm. 56, 250-256.

15