Responses of Aphis fabae Scop (Hom., Aphididae) and its
steno-phagous predator Cheilomenes propinque Muls.
(Col., Coccinellidae) to neem seed water extract
By
Mohammed Abdalla Suliman Mohammed
B.Sc. Agric. (Honours) Sci. (2002).
University of Gezira
A Thesis Submitted in Partial Fulfillment of the Requirements
for the Degree of M. Sc. in Crop Protection
Supervisor:
Dr. Hamadttu Abdel Farag El Shafie
Department of Crop Protection
Faculty of Agriculture
University of Khartoum
September - 2005
1
DEDICATION
To my father
To my dear mother
To my grand mother (Haja Sakina)
To my brothers and sisters
To my teachers and friends
With love and respect
`É{tÅÅxw
Acknowledgements
2
First of all, I render my gratitude and praise to the Almighty “Allah”.
I wish to express my sincere gratitude to Dr. Hamadttu Abdel Farag
Elshafie, for his guidance continuous encouragement, and efforts to solve
the problems arised during the accomplishment of this research.
Iam deeply grateful to Dr. Abdelwahab Hassan for choosing a
suitable design for the experiment and Dr. Salah Hamed Elturabi for his
help with the statistical analysis.
I would like to extended my thank to Dr. Moutassim Mohamed
Khalfallah (Environment and Natural Research Institute)
I acknowledge with thanks the help rendered to me by my colleagues
and staff members of the Department of crop protection, Faculty of
Agriculture, University of Khartoum.
Finally, special thanks are extended to my family and my friends
ABSTRACT
3
Two green house experiments were conducted at Shambat area
to investigate the systemic and contact effect of neem seed kernel
aqueous extracts on the bean aphid, Aphis fabae Scop. and their stenophagous Coccinellid predator Cheilomenes propinque (Muls.).
In the first experiment, different concentrations of neem seed
kernel aqueous extract (25g, 50g and 75g/liter of water), and
insecticide Actara® (1g per 1.25 litter of water) were tested. Three
methods of application: drenching in the soil, spraying on the foliage
and topically on the aphid body were attempted.
In the second experiment, the same concentrations of neem and
the same insecticides were applied topically (direct spraying) on the
different stages of C. propinque (Muls)(eggs, larva, adults).
The results indicated that, Actara® was the most effective
product to control aphids and kill their predator C. propinque (Muls.),
compared with neem seed aqueous extracts.
The results in all experiments indicated that the effect of neem
seed kernel aqueous extract was effective in the first 3–4 days of the
experiment. Also during these days, the high concentration of neem
gave high mortality.
The results have also shown that the neem products were less
effective on the C. propinque (Muls.) than aphids.
4
‫ﺑﺴﻢ اﷲ اﻟﺮﺣﻤﻦ اﻟﺮﺣﻴﻢ‬
‫ﺨﻼﺼﺔ ﺍﻷﻁﺭﻭﺤﺔ‬
‫ﺃﺠﺭﻴﺕ ﺘﺠﺭﺒﺘﺎﻥ ﺩﺍﺨل ﺍﻟﺼﻭﺒﺔ ﺍﻟﻤﺤﻤﻴﺔ ﺒﻤﻨﻁﻘـﺔ ﺸـﻤﺒﺎﺕ ﻟﺩﺭﺍﺴـﺔ ﺍﻟﻔﻌـل ﺍﻟﺠﻬـﺎﺯﻱ‬
‫ﻭﺒﺎﻟﻤﻼﻤﺴﺔ ﻟﻤﺴﺘﺨﻠﺼﺎﺕ ﺒﺫﻭﺭ ﺍﻟﻨﻴﻡ ﻋﻠﻰ ﺤﺸﺭﺓ ﻤـﻥ ﺍﻟﻔـﻭل ﺍﻟﻤـﺼﺭﻱ ‪Aphis fabae Scop‬‬
‫ﻭﻤﻔﺘﺭﺴﻬﺎ ﺨﻨﻔﺴﺎﺀ ﺃﺒﻭ ﺍﻟﻌﻴﺩ )‪.Cheilomenes propinque (Muls‬‬
‫ﻓﻲ ﺍﻟﺘﺠﺭﺒﺔ ﺍﻷﻭﻟﻰ ﺃﺴﺘﺨﺩﻤﺕ ﺜﻼﺜﺔ ﺘﺭﻜﻴﺯﺍﺕ ﻤﻥ ﻤﺴﺘﺨﻠﺹ ﺒﺫﺭﺓ ﺍﻟﻨﻴﻡ )‪ 25‬ﺠـﺭﺍﻡ‪50 ،‬‬
‫ﺠﺭﺍﻡ‪ ،‬ﻭ ‪ 75‬ﺠﺭﺍﻡ ﻟﻜل ﻟﺘﺭ‪/‬ﻤﺎﺀ( ﻭﻤﺒﻴﺩ ﺃﻜﺘﺎﺭﺍ ®‪ Actara‬ﺍﻟﺤﺸﺭﻱ ﺒﺘﺭﻜﻴﺯ )‪ 1‬ﺠﺭﺍﻡ ﻟﻜـل ‪1.25‬‬
‫ﻟﺘﺭ ﻤﺎﺀ( ﻭﻗﺩ ﺃﺴﺘﺨﺩﻤﺕ ﺜﻼﺜﺔ ﻁﺭﻕ ﻟﻤﻌﺎﻤﻠﺔ ﺤﺸﺭﺓ ﺍﻟﻤﻥ‪ :‬ﺍﻷﻭﻟﻰ ﺍﻟﺭﺵ ﻋﻠﻰ ﺍﻟﺘﺭﺒﺔ ﻭﺍﻟﺜﺎﻨﻴﺔ ﺍﻟﺭﺵ‬
‫ﺍﻟﻤﺒﺎﺸﺭ ﻋﻠﻰ ﺍﻟﺠﺯﺀ ﺍﻟﺨﻀﺭﻱ ﻤﻥ ﺍﻟﻨﺒﺎﺕ ﻭﺍﻟﺜﺎﻟﺜﺔ ﺍﻟﺭﺵ ﺍﻟﻤﺒﺎﺸﺭ ﻋﻠﻰ ﺠﺴﻡ ﺍﻟﺤﺸﺭﺓ‪.‬‬
‫ﻓﻲ ﺍﻟﺘﺠﺭﺒﺔ ﺍﻟﺜﺎﻨﻴﺔ ﺃﺴﺘﺨﺩﻤﺕ ﻨﻔﺱ ﺍﻟﺘﺭﻜﻴﺯﺍﺕ ﺍﻟﻤﺨﺘﻠﻔﺔ ﻟﻠﻨﻴﻡ ﻭﻨﻔﺱ ﺍﻟﻤﺒﻴﺩ ﺍﻟﺤﺸﺭﻱ ﺒﻁﺭﻴﻘﺔ‬
‫ﺍﻟﺭﺵ ﺍﻟﻤﺒﺎﺸﺭﺓ ﻋﻠﻰ ﺃﻁﻭﺍﺭ ﺨﻨﻔﺴﺎﺀ ﺃﺒﻭ ﺍﻟﻌﻴﺩ ﺍﻟﻤﺨﺘﻠﻔﺔ )ﺍﻟﺒﻴﻀﺔ ‪ ،‬ﺍﻟﻴﺭﻗﺔ‪ ،‬ﺍﻟﺤﺸﺭﺓ ﺍﻟﻜﺎﻤﻠﺔ(‪.‬‬
‫ﺃﻭﻀﺤﺕ ﺍﻟﻨﺘﺎﺌﺞ ﺃﻥ ﻤﺒﻴﺩ ﺃﻜﺘﺎﺭﺍ ﺍﻟﺤﺸﺭﻱ ﺃﻜﺜﺭ ﻓﻌﺎﻟﻴﺔ ﻤﻘﺎﺭﻨﺔ ﺒﻤﺴﺘﺨﻠـﺼﺎﺕ ﺍﻟﻨـﻴﻡ ﻓـﻲ‬
‫ﻤﻜﺎﻓﺤﺔ ﺤﺸﺭﺓ ﺍﻟﻤﻥ ﻭﺍﻟﻘﻀﺎﺀ ﻋﻠﻰ ﻤﻔﺘﺭﺴﻬﺎ ﺃﺒﻭ ﺍﻟﻌﻴﺩ‪ .‬ﻜﺫﻟﻙ ﺃﻭﻀﺤﺕ ﺍﻟﻨﺘﺎﺌﺞ ﻓﻲ ﻜل ﺍﻟﺘﺠﺭﺒـﺔ ﺃﻥ‬
‫ﺘﺄﺜﻴﺭ ﻤﺴﺘﺨﻠﺼﺎﺕ ﺍﻟﻨﻴﻡ ﺒﻜل ﺘﺭﻜﻴﺯﺍﺘﻬﺎ ﺇﻨﺤﺼﺭﺕ ﻓﻲ ﺍﻟﺜﻼﺜﺔ ﺇﻟﻰ ﺃﺭﺒﻌﺔ ﺃﻴﺎﻡ ﺍﻷﻭﻟﻲ‪ .‬ﻜﻤﺎ ﺃﻭﻀﺤﺕ‬
‫‪5‬‬
‫ﺍﻟﻨﺘﺎﺌﺞ ﺨﻼل ﺍﻟﻴﻭﻡ ﺍﻟﻭﺍﺤﺩ ﺃﻥ ﺍﻟﺘﺭﻜﻴﺯ ﺍﻟﻌﺎﻟﻲ ﻤﻥ ﻤﺴﺘﺨﻠﺹ ﺍﻟﻨﻴﻡ ﻜﺎﻥ ﻫﻭ ﺍﻷﻓﻀل ﺒﻴﻥ ﻤﻌـﺎﻤﻼﺕ‬
‫ﺍﻟﻨﻴﻡ ﺤﻴﺙ ﺃﻋﻁﻲ ﻤﻌﺩل ﻤﻭﺕ ﻋﺎﻟﻲ‪.‬‬
‫ﻜﺫﻟﻙ ﺃﻭﻀﺤﺕ ﺍﻟﻨﺘﺎﺌﺞ ﺃﻥ ﺘﺄﺜﻴﺭ ﻤﺴﺘﺨﻠﺹ ﺍﻟﻨﻴﻡ ﻜﺎﻥ ﻀﻌﻴﻔﹰﺎ ﻋﻠﻰ ﺍﻟﻤﻔﺘـﺭﺱ ﺃﺒـﻭ ﺍﻟﻌﻴـﺩ‬
‫ﺒﺎﻟﻤﻘﺎﺭﻨﺔ ﺒﺤﺸﺭﺓ ﺍﻟﻤﻥ‪.‬‬
‫‪6‬‬
LIST OF CONTENTS
Page
Dedication………………………………….…………………………………….……
i
Acknowledgements ……………………………….………………………….……
ii
Abstract …………………………...……….…………………………………….……
iii
Arabic Abstract ……………………………………………………………….……
iv
List of Contents ……………………………………………………………….……
v
List of Tables and Figures ….….…….…………………………………….……
ix
List of Plates ………………………………...……………………………….……
x
List of Abbreviations……………………………………………..………….……
xi
CHAPTER ONE: INTRODUCTION …….………………..…………….…
1
CHAPTER TWO: LITERATURE REVIEW…….……..…………….…
3
2-1-1 Classification of aphids…………..…………………..……………….…
3
2-1-2 Nature of damage………………………………………………………..…
3
2. The control measures adopted against aphids…….……………….…
4
2.1. Conventional method….………………….…………………………….…
4
2.1.1. Chemical control….…………………………..……………………….…
4
2.1.2. Biological control…….………………...……………………………….…
4
7
2.2. The use of botanical (Neem tree) in control of aphids……...…
5
2.2.1. Chemical constituents of neem…….……………………...……….…
5
2.2.2. The mode of action of azadirachtin…….……………………….…
5
2.2.3. The side effect of neem kernel Aqu- Ext on the aphids….…
6
2.3 The Coccinellids…….………………………...…………………………….…
8
2.3.1. Classification of C. propinque (Muls)…….………………….…
8
2.3.2. Population dynamic of C. propinque (Muls) …….……………
8
2.3.3. The efficiency of coccinellids in checking aphid popula-tion
…….…………….…………….………………….…………………………………
9
2.3.4. Side–effect of neem seed Aqu-Ext on the Coccinellids
10
CHAPTER THREE: MATERIALS AND METHODS…….…
13
3.1. Aphids culture….……………………….………………………………….…
13
3.2. Collection and rearing of Cheilomenes propinque (Muls) …….
13
3.3. Preparation of test materials……..…………………………………….…
15
3.3.1. Neem seeds kernel powder (NSKP) …….…………………………
15
3.3.2. Preparation the aqueous- extract…….………………………………
15
3.4. Synthetic insecticide…….………………..……………………………….…
15
3.5. Equipment and spraying technique…….……………..…………….…
16
8
3.6. The effect of neem aqueous-eztract on the Aphis fabae (Scop)
…….………………………………………………………..…………….…
16
3.6.1 Soil drenching experiment…….…………………………………….…
16
3.6.2. Foliar application experiment…….……………………………….…
20
3.6.3. Topical application experiment …….…………………………….…
20
3.7. The efficacy of neem aqueous extract on the Cheilomenes
propinque (muls) …….…………………………………………..……………….…
21
3.7.1. Egg stage…….……………………………………….…………………….…
21
3.7.2. Larval stage…….………………………………………………………….…
22
3.7.3. Adult stage…….………………………………….……………………….…
22
3.7.4. Feeding of C. propinque (Muls) on aphids treated topically……
22
CHAPTER FOUR: RESULTS……………….……………………………
25
4.1.1. The effect of NSK Aqu-Ext, applied as soil drenching, on
aphids (drenching experiment) …….………………………………….…
25
4.1.2. The effect of NSK Aqu-Ext sprayed on the faba bean
plants…….…………………………………………………….…………………….…
27
4.1.3. The effect of NSK Aqu-Ext sparyed topically on the
aphids…….………………………………………………………...………………….…
27
4.2.1. The effect of NSK Aqu-Ext on the Egg hatchability of C.
propinque (Muls) 48 hours after treatment .…………………. ……….…
9
30
4.2.2. The effect of NSK Aqu-Ext on the Aduts C. propinque Muls.
30
4.2.3. The effect of NSK Aqu-Ext on the 3rd, larval instar…….…
30
4.2.4. The feeding of C. propinque (Muls) on aphids treated topically
with NSK Au- Ext………………………………………………….…
32
CHAPTER FIVE: DISCUSSION …….……………………………….…
36
5.1. General…….………………………………………………..………………….…
36
5.2. The effects of NSK Aqu-Ext on aphids…….………………….…
36
5.3. The effects of NSK Aqu-Ext and Actara® on the different stages
of the Coccinellid Cheilomenes propinque (Muls) …….…
38
5.4. Conclusions and recommendations……………………………….…
39
REFERENCES…….……………...………………..…………………………….…
41
APPENDICES …….…………………………...……………………………….…
51
10
LIST OF TABLES & FIGURES
Table Title
1.
Page
Survival and corrected mortality (CM) of A. fabae (20
insects) fed on seedling of faba bean treated with NSWE
(soil application)……………………………………………………………
2.
26
Survival and corrected mortality (CM) of A. fabae (20
insects) after foliar application of NSWE on faba bean (post
infestation). ………………………….………………………………………
3.
Survival and corrected mortality (CM) of A. fabae (20
insects) topically sprayed with NSWE……………………………
4.
29
The effect of topical application of NSK Aqu-Ext on the egg
hatchability of (C. propinque Muls) 48 hours after treatment
5.
28
31
Survival and corrected mortality (C.M) of C. propinque
adults (10 insects) after direct spraying with, neem seed
water extract (NSWE)…….………………………………………………
6.
33
Survival and corrected mortality (C.M) of 3rd larvae (10
insects) of C. propinque after direct spraying with, neem
seed water extract (NSWE)……………………………………………
7.
34
Survival and corrected mortality (C.M) of C. propinque
adults (10 insects) fed on aphids (A. fabae) treated topically
with, neem seed water extract (NSWE)………………….…………
11
35
LIST OF PLATES
Plate
Title
Page
1.
Aphid and Coccinellids rearing culture…………………………
14
2.
The sprayer used in experiments……….…………………………
17
3.
The experimental unit…………………..….…………………………
19
4.
The test stages of C. propinque …………………………...………
24
12
LIST OF ABBREVIATIONS
ANOVA
Analysis of variance
ARC
Agricultural Research Center
AZA
Azadirachtin
CM
Corrected mortality
cm
Centimeter
D.f.
Degree of freedom
Ec
Emulsifiable concentrate
Fcal
The ratio of ms of treatment to ms of experimental error
Fig.
Figure
IPM
Integrated pest management
L
Liter
NSO
Neem seed oil
NSK
Neem seed kernel
NSK Aqu-Ext
Neem seed kernel aqueous extracts
NSKP
Neem seed kernel powder
NSWE
Neem seed water extract
Ss
Sum of square
13
CHAPTER ONE
INTRODUCTION
Broad bean (Vicia faba L.) has many names but the name faba bean
has recently been widely used. It is considered as good source of protein,
carbohydrates and fiber (El Tinay et al., 1993.).
Faba bean is subject to attack by several insect pests and disease from
the early development phase to the post harvest time. The main pest that
attacks faba bean in the early development phase is aphid.
Aphids are pests of world-wide importance and they cause heavy
crops losses by feeding on plant sap and transmitting viruses as vectors. In
addition, they also secrete copious amount of honey dew which attracts
sooty mould. Although aphids are readily killed by various insecticides, yet
they are not easily controlled because they multiply rapidly and many
species feed on the lower side of the foliage and therefore not easily
reached by sprays or dusts (Frohlch and Rode Wald, 1963). Aphid, are also
known to develop resistance against insecticides rapidly.
Therefore, it is necessary to find alternative methods of control
preferably with agents non- toxic to humans and less hazardous to the
environment.
The natural enemy of aphids [C. propinque (Muls)], as an example
taken in this study plays a fundamental role in integrated pest management
program, Coocinellids were found to be the common predators attacking
cereal aphids in spring cereal fields (Dean, 1974, Jones, 1972). They are
also important predators of aphids in several field crops (Nevenschewander
et al. 1976, Kring et al. 1985. Rice and Wide 1988).
14
Obvious pollution problems in the environment, and the toxic effects
of synthetic chemicals on non-target organisms have prompted investigation
on exploiting pesticides of plant origin. Natural plant products and their
analogues are important source of new agricultural chemicals used in the
control of insect pest (Cardllina 1988, Emosairue and Ukeh 1996 Gulter,
1998, Schmutterer and Huber, 2005).
Recent studies have shown the importance of natural chemicals as
possible source of non- phytotoxic, systemic, and easily biodegradable
alternative pesticides (Singh, 1994; Qasem and Abou-Blan, 1996, Mason
and Mathew, 1996). Furthermore, pesticides of plant origin are cheap,
readily available and cost effective in developing countries.
Many pesticides of plant origin have been used as pest control agents.
About 2000 plant species were reported to posses pest control properties,
out of which very little are being used for insect control purposes. Products
from the neem tree (Azadirchata indica A Juss) proved to have promising
effects against pests. Mortality, antifeedent, repellency development and
oviposition inhibitions are some of the impacts of the neem on insects.
This studied was taken under the green house conditions with the
main following objectives.
To evaluate the contact and systemic action of the extracts from the
neem seed kernel (NSKWE) on faba been aphid Aphis faba (Scop).
To elucidate the negative impact of neem, if any on the
developmental stages of the steno-phagous aphid, on predator [Cheilomenes
propingue (Muls)].
15
CHAPTER TWO
LITERATURE REVIEW
2-1-1 Classification of aphids:
Aphids belong to the order: Hemiptera, Sub order: Homoptera, super
family: Aphidoidea and family Aphididea (Imms, 1957).
2-1-2 Nature of damage.
Aphids are pests of world-wide importance and they cause heavy
crop losses by feeding on plant sap and transmitting viruses as vectors.
Inaddition, they also secrete copious of honey dew which attracts sooty
moulds.
Many aphids species are vectors of viral disease and they can cause
serious damage at low densities. Some species of aphids such as the green
peach aphid Myzus persicae (Sluzer) which is vector of many viral disease
has developed resistance to insecticides (Fernhurst et al., 1989).
The virus of bean yellow mosaic (BYMV) is transmissible by sap
inoculation and by aphids in a non- persistant manner. The well known
aphid vectors include Aphis craccivora, Aphis fabae, A. Pisum and Myzus
persicae (Quantz, 1954; Nitzany and Cohen, 1962; Evans 1973).
Black bean aphids (Aphis fabae Scop) is commonly recognized as
pest of number of crop in Europe. In Poland this species has been recorded
as a harmful pest of broad bean, field bean and sugarbeet (Goszczyhski et
al., 1992, Hurej, 1991). In field bean Vicia faba, yield losses due to this
species were found to depend on the timing and intensity of colonization
16
(Hins et al, 1981). In the Sudan, a number of aphid species were reported
from agricultural research stations. They include, The wheat aphids
Schizaphis graminum (Rondani), Rhopalosiphum maidis (Fitcg), the cotton
Aphid gossypii (Glover), the cowpea aphid Aphis craccivora Koch, the
sorghum aphid Aphis sorghi (Theo); the milk weed or the oliander aphid,
Aphis nerii Fonscolombe; and Aphis lutescents Monell.
2. Control measures adopted against aphids:
2.1. Conventional methods:
The conventional methods of insect pests control used in Sudan are
applicable also for aphid control viz. chemical control use of resistant
varieties, good crop husbandry…etc.
2.1.1. Chemical control:
Mudathir (1971) found that various insecticides gave a good control
and effectively reduced aphid population. These were mostly organo
phosphorus compounds namely Folimat®, 80% Primor® 25%, vamifen®
25%, Thimulion®, Anthio ® 25%, Ekatin® 25%, Dimethoate® 32%,
cytrolane® 50%, Metasytox® 50%, Lannate ® 90%, Cp5 11%, Trihion 4E,
Tamarn® 50 % and Bidrin® 24%.
2.1.2. Biological control:
Cereal aphids as example are attacked by wide range of parasitoids,
predators and fungal pathogens. Bacteria and viruses have not been reported
(Elamin, 1975). Coccinellids were found to be the common predators
attacking cereal aphids in spring cereal fields (Dean, 1974; Jones, 1972).
17
In the Sudan Gezira, the following aphid predators were reported and
found to play an important role in aphid control: the beetle Coccinella
undecimpunctata. (L), the green lace wing (GLW) Chrysoperla
(=chrysopa) zastrowi Ebson-petrson, C. paudica Naras, the lady bird
Cheilomenes
propinque
propinque
(Mulsant)[=Cydonia
propinque
(Mulsant)], the ladybird Hippodamia variegata (Goeze), the syrphid
Ischiodon aegyptium Wieden mann, and the lady bird Hyperapsis spp. (
Munir et al. 1992).
2.2. The use of botanical (Neem tree) in the control of aphids:
Neem is a forest plant in Sudan .It had been introduced from India. It
is now extensively grown throughout the country as shade tree in public
parks, street and residential areas. It is also used in shelter belts in dry area.
The origin of neem tree is south-east Asia, where it has been used in
traditional medicine, production of insecticides, lamping oil, soap from
manufacturing the margosa oil pressed from its seed kernels which contain
up to 40% oil (Siddig, 1991).
2.2.1. Chemical constituents of neem:
All of the well characterized compounds identified in Neem tree
belong to the class triterpenoids. These include Azadirachtin, Salannin,
Salannol, Salannol acetate, 3-deacetyl-salannin, nimbin and nimbidin and
nimbolid ( Schmutterer and Zebitz, 1984).
2.2.2. The mode of action of azadirachtin:
Within azadirachtin molecule, the declain fragment is responsible for
the insect growth regulation and development effects observed, while the
18
hydroxylfuran fragment causes the anti-feedant effects more widely
observed among target species (Aldhous, 1992). The IGR and anti-feedant
effects of azadirachtin are independent of each other, but both remain
relative to concentration (Koul & Isman 1991).
Schmutterer (1990) suggested that azadirachtin modifies the
developmental programs of insects by influencing hormonal systems,
especially that of ecdysone. The effects of azadirachtin are both dose and
time dependent, prevent both ecdysis and apolysis, and can cause insect
death.
2.2.3. The side effect of neem kernel Aqu- Ext on the aphids:
Neem seeds and leaves extract gave better control of Aphis gossypii
on cotton and potato, compared to the standard insecticides Sevin + Anthio
and Endosulfan (Venkatrtan et al., 1987 and Sidding, 1991). Elshafie
(2001) investigated the effect of three different neem formulations
including NeemAzal T/S on Aphis gossypii, Jacobiasca lybica and Bemisia
tabaci damaging potato and egg plants. He reported that neem preparation,
particularly NeemAzal gave results which were comparable with
Sumicidin- Neem seed water extracts gave good result against the flea
beetle, Podagrica spp., white fly Bemisia tabaci Genn, cotton jassid
Jacobiasca lybica Pruthi, Aphis spp. and spying boll worm Earias insulana
Boisd on okra crop in the Sudan (Siddig, 1991).
In the Sudan, neem seed and leaves water extracts at 1kg /40L of
water,repelled foliage pest of potato i.e Bemisia tabaci, Jacobiasca lybica
and Aphis gossypii increased yield by 0.5 tones/ha. Therefor, a pest
19
management program should include neem treatments for combating potato
pests in Sudan (Siddig, 1987).
Tertiary–methyl–butyl ether extract and methonolic extract of
neem seed kernel (MENSK) were toxic to the first instar of Aphis pisum
and Aphis fabae (Schauer, 1984). Dimetry and Schmidt (1992) counted
more apetrous females of the bean aphid, Aphis fabae on plants in a choice
test where as “NeemAzals” especially after application of higher
concentrations (0.1 – 2%) deterred the aphids.
A mixture of plant extract including neem at (1%, 4%, 10%) was
highly repellent to the bean aphids, Acyrthosiphon pisum (Hunter and
Ullman, 1992). Molting failure was observed in young aphids, Aphis pisum,
exposed to bean plants sprayed with NSKEs (Schauer, 1984). Persistence of
toxic neem oil to aphids was limited to a bout 3 to 7 days outdoors and it
remained active for longer periods on plant in green house ( lowery , 1992).
Lowery (1992) observed that the third instar of Myzus persica and
Nasonovia ribisnigri exposed to 5% neem oil molted successfully to adults,
but they were generally much smaller in size than those in the control.
Many homopterous and aphids are highly sensitive to neem products
(Elshafie, 200l). Neem based insecticides can be effective in controlling
aphid (Schauer 1984, schmutterer, 1990 and Nisbet et al., 1994).
2.3 The Coccinellids:
2.3.1. The Classification of Cheilomenes propinque (Muls):
20
The Coccinellid Cheilomenes propinque (Muls) belong to the order
Coleoptera, suborder: polyphaga, superfamily Cucujoidea and the family
Coccinellidae Imms, 1957) .
C. propinque is a polyphagous Coccinelld with a wide range of prey
acceptance. It has been reported by many workers to prey on different aphid
species and other agronomic and horticultural pests in Africa. In the sudan
this predator was previously reported to feed on Melaphis sacchari
(Zehnter) and Myzus utricae (Kalt) Cited by Bashir (1968). Pea cock (1913)
and Lamborn (1914), also reported that A. gossypii infesting cotton in south
Nigeria was kept well in check by C. propinque (Ibrahim, 1988).
2.3.2. Population dynamic of C. propinque (Muls).
Adults and larvae of C. propinque were common in lucerne fields
during April to October. They seemed to migrate from by nearly sorghum
fields, they where already present in large numbers during January-April
and when M. sacchari started to decline (Basher, 1968). Similar
observations were made by Mohamadein (1979), but he added that C.
propinque was the least prevalent in lucerne fields. Bashir (1968), showed
that this predator is of less important in the natural control of A. craccivora.
The sensitivity of C. propinque to pesticides was reported by Mohamadein
(1979). The two species almost disappeared from the sprayed fields
until the end of September under natural conditions, the efficiency of C.
propinque as a predator of the cowpea aphid may be affected by inter and
intra specific competition ….etc. However, the laboratory data are
supportive of considering C. propinque as an important biotic control agent
of the cowpea aphid (Ofuya, 1986).
21
2.3.3. The efficiency of Coccinellids in checking aphid population:
The predators (Coccinellids) play an important role in the control and
reduction of aphids internationally. The following reports are of pertinent.
Kenchaiah and Porte (1989) reported that field surveys were
conducted in several districts of Kanataka in India during Kharif (autumn)
and rabi (spring) seasons of 1984 – 1989 to determine the major insect pests
of groundnut. The average incidence of aphids during the Kharif and rabi
seasons was moderate (16-35 %), and low (1–5%), respectively.
The maximum incidence of aphid population on groundnut was
recorded between July and September, 1987–1988. Increases in the number
of aphids were controlled by the Coccinellid predators Coccinellia sp, and
Menochilus sexmoculatus Fabricius (Cheilomenes sexmoculata F.). In the
Harihar and Chitradurga districts, 5–10 larvae of aphid populations during
mild weather.
The predation efficiency of the Coccinellds Coccinella transversalis
Fab, and M. sexmaculatus against 3 species of aphids (Aphid faba Scop., A.
Gossypii, and A. nerrii) were discussed in relation to prey preference, and
bioenergeties of the predators. C. sexmeculata tended to exhibit lower
predatory efficiency than C. transsversalis. Feeding preference, longevity
and reproductive potential, beside predatory efficiency of these predators
showed significant correlation with the aphid prey quality (Babu and
Ananthakmishan, 1993).
The relationship between A. craccivora on groundnut and its
predators was studied in Gujarat, India, 1981. The predators collected were
22
identified as H. variegata, C. Septempunctata, and M. sexnaculatus. An
infestation index was calculated by dividing the number of plants showing
various extents of damage by the total number of plant examined. A
positive correlation was observed between aphid index and population of
active states of the predators during March, while in April the aphid
population decreased with increased abundance of the Coccinellids. The
predators population declined by the 1st week of may, most of them having
migrated to other areas, due to decrease in prey abundance. It was
concluded that application of insecticides is unnecessary, if the Coccinellds
are present in the groundnut crop in Summer (Butani and bharodia, 1984).
2.3.4. Side–effects of neem seed Aqu-Ext on the Coccinellids.
The side effects on beneficials and other non- target organisms is
important aspect concerning the use of neem products in integrated
production and organic farming.
Neem products can cause some mild effects on non- target insects
and therefore, recommended for pest control especially in IPM
(Schmutterer, 1995). Experiments with adults of the Coccinellid beetle
(Coccinella septempunctata) kept on neem oil did not show increased
mortality or reduction of fecundity when compared with untreated control,
but the metamorphosis of the larvae was interrupted (Schmutterer, 1981).
Lowery and Isman (1995), reported that the number of larvae of
predacious Cecidoryiid’ was reduced in the field after application of neem
seed extract and neem oil as compared with the control.
23
Neem was found to be harmless or slightly harmful to natural
enemies like predators and parasites. Predtors spider. L pseudonulata was
not affected even by 50g neem oil per individual (Saxena, et al., 1984).
Lowery and Isman (1995) reported that the edosion of adult Coccinellids
(Coccinella undecimpunctata) and the syrphid (Eupeodes fumipennis) was
negatively affected.
El shafie (2000) reported that the application of neem water extract,
neem oil and NeemAzal did not reduce the population of the aphid
predators Scymnus spp. He observed that the reduction in the population of
the Coccinellid was correlated to the reduction of the prey and was not due
to the dirct effect of neem preparation.
Neem research has led to the assumption that fewer side-effects on
beneficial organisms could be expected when neem is compared with the
numerous pesticides with strong contact and neuro –toxic mode of action
against the active Coccinellids. The application of neem formulation in the
laboratory, when used with the same concentration as in the filed, lead to a
strong side-effect on beneficial organisms than the field, where, they are
often not hit by sprays and where relatively rapid degradation of active
ingredients takes place. Spraying of neem oils against the sorghum aphid
Melanoaphis sacchari in the field trials at concentration of 0.1, 0.2, 0.5, and
1.0, did not harm the larvae of Coccinellids or the larvae of syrphids
(Schmutterer, 1995).
Green house experiments were conducted at Shambat area to
investigate the systemic and contact effects of different neem formulations
on the Coccinellid Hippodamia variegata (Goeze). The results indicated
24
that Sumicidin® 20% Ec (0.5mL/L) was the most effective on the different
developmental stages of the Coccinellids specially, the larvae compared to
the neem products, while neem seed water extract (2.5%) showed the least
effect compared to celatlor® (4mL/L). It is found that feeding of
Coccinellid on aphid treated topically was the most effective treatments
compared to direct spraying and aphid treated systemically. Also feeding of
Coccinellid on aphid treated systemically gave weak effect as compared
with other treatments (Hamad, 2003).
25
CHAPTER THREE
MATERIALS AND METHOD
3.1. Aphids culture:
Four plastic cups, 30 cm in diameter, were sown with faba bean
(Vicia faba L.) cultivar Hudieba 93 and every cup was kept inside small
cages (62.5×45×45cm) made of wooden frames and wire-mesh at the sides
and covered with muslin cloth at the top (plate 1). Adults of the Black bean
aphid [Aphis faba (Scop)] were obtained from the Demonstration farm of
the Faculty of Agriculture, University of Khartoum in Shambat from
untreated faba bean plants. The insects were collected in Petri-dishes with
the aid of fine camel hair brush and were used to infest the faba bean plants.
Insects were left to reproduce and develop for many generation before
being used in the different experiment.
3.2. Collection and rearing of Cheilomenes propinque(Muls):
Stock culture of C.vcina (Muls) established from pupal stages
collected from the medicag sativa grown in the farm of the Faculty of
Agriculture, University of Khartoum in Shambat. The collected pupae were
put in Petri-dishes. Cages (62.5×45×45cm) were used to rear the
Coccinellid in the green house (plate 1). The Adult insects were f ad libitum
on aphid reared on faba bean plants. They were left to reproduce and kept
as test insects until the end of all experiment.
26
Plate 1. Aphid and Coccinellids rearing cage
27
3.3. Preparation of test materials:
The collection of neem seeds, the preparation of powder, the
extraction and methods of storage were carried out according to those
described by Dreyer (1984), Ermal et al (1984), Schmutterer (1988) and
Siddig (1990).
3.3.1. Neem seeds kernel powder (NSKP):
Mature fallen neem fruit berries were collected in Shambat area. The
seeds were washed and crushed lightly by stone to break the seed coat
which were then separated from the broken seed kernel by winnowing,
Pestle and motor were then used to mill the kernels into fine powder.
3.3.2. Preparation the aqueous- extract:
The neem seeds kernel powder prepared as above, was put in tap
water in plastic bucket and stirred thoroughly with a piece of wood for 10
min, and allowed to stand overnight. After 24 hours the mixture was reagitated for 10 min and filtered, using a light cloth, to prevent blocking of
the sprayer’s nozzle.
3.4. Synthetic insecticide:
Actara® neonicotinoid was used at recommended dose (1g/1.25 liter
water). This was to allow the comparison of its side–effects with that of
aqueous-extract of neem seeds kernel. This insecticide was chosen because
it has been proved as effective agent against 160 pest species in 115 crops.
Moreover, it has a broad action against a wide spectrum of sucking and leaf
–feeding pests.
28
3.5. Equipment and spraying technique:
A plastic hand sprayer (0.5 liter) was used to apply the test chemicals
in all experiment. The pressure was maintained by hand continuously
during spraying (plate 2) . This sprayer gives very small droplets and hence
a good surface coverage. The sprayer was washed thoroughly with water
after each concentration used. The preparations were poured into the
sprayer using a glass flask and filtered through piece of fine cloth.
3.6. The efficiency of neem aqueous-extract on the Aphis fabae (Scop):
Three experiments were carried out in the Green house of the Faculty
of Agriculture, University of Khartoum, Shambat “15°. 40 °North latitude,
longitude 32°. 32´ and 280 meters above sea level” for the purpose of
assessing the systemic and contact efficacy of neem seed kernel and time of
application in controlling the black bean aphid Aphis fabae (Scop). The
experiments were carried out during the period from November to
December 2004.
3.6.1. Experiment–I (Soil drenching):
This experiment was carried out when the faba bean (Vicia faba L)
seedling reached the four leaves stage in winter season to study the systemic
effect of the aqueous extract of NSKP on the faba bean aphid when applied
by drenching the soil before infestation (pre- infestation application).
29
Plate 2. The sprayer used in the experiments
30
Twenty plastic pots 10 cm wide at the top and half-filled with sterile
soil, were planted with faba bean (Vicia faba L) Cultivar (Hudieba 93).
Three seeds were put in each pot and after germination, the seedlings were
thinned to two plants per pot. Each pot was then covered with mulslin cloth
(transparent cloth) to allow the access of plants to sun light and to prevent
contamination of test plants with any other insect (plate 3). The pots were
treated with the different treatments before aphids were transferred on them.
Each treatment was replicated four times. The infestation was carried out by
placing 20 aphids on the plants in each pot. The aphids transfer was carried
out 24 hours after treatments (post-infestation application). The soil
drenching of each pot was carried out by spraying the soil by 20 ml. The
control pots (untreated) were drenched with water only. The treatments
were as follows:
1-
25 g NSKP/Liter water.
2-
50 g NSKP/Liter water.
3-
75 g of NSKP/Liter water.
4-
Actara® neconicotinoid used at recommended Rate (1g/1.25Liter
water).
5-
Control (water only).
The number of live insects (Aphids) were counted in each pot every 24
hours for 7 days.
31
B4
Plate 3. The experimental unit
32
3.6.2. Experiment II (Foliar application):
This experiment was performed during the same period when
seedlings reached the four leave stage in the winter season. The specific
objective of the experiment was to study the effect of the NSK aqueous
extract on faba bean aphids when applied by spraying the plant before
infestation. Twenty plastic pots (10 cm) at the top and each pot semi-filled
with sterile soil, were planted with faba bean (Vicia faba) Cultivar
“Hudieba 93” three seeds per pot.The seedlings were thinned to two plant
per pot. Each pot was coverd with muslin cloth, the pots were treated with
the different treatments before they were infested. Each treatment was
replicated four times. The infestation was carried out by placing (20) aphids
on the plants in each pot. After 24 hours, the aphids were placed on the
plants (post infestation application). A plastic hand sprayer as described
above was used. The foliar parts of the plants on each pot was sprayed
carefully to ensure good coverage of the shoot part of the plants. The
control plots (untreated) were sprayed with water.The same treatments as
mentioned in the first were used. The number of live insects (aphids) were
counted and recorded in each pot every 24 hours for 7 days.
3.6.3. Experiment – III (Topical application):
This experiment was performed consonantly with the above two
experiments, in the winter season to study the effect of NSK aqueous
extract when applied by direct spraying (Topical application). Twenty Petridishes were arranged in a completely randomized design (CRD), twenty
aphids were placed in every petri-dishes and were then treated with
different treatments. The insect was fed with a parts of faba bean plants
33
daily. Each treatment was replicated four times. The same treatments, as
mentioned in the above experiments were used. The number of live insect
(aphids) were counted every 24 hours for 7 days.
3.7. The side-effects of neem aqueous extract on the Cheilomenses
propinque (muls):
Four experiments were carried out, for the purpose of assessing the
effect of the neem seed kernel applied topically on the natural enemies C.
propinque (Muls). All experiments were carried out during the period from
the November to December 2004. The different stages of the C. propinque
(Muls)(Plate 4) were sprayer with neem seed water extract (NSWE) and the
Actara® insecticide.
3.7.1. Egg stage:
Ten eggs in a petri-dishes containing filter papers were sprayed to
determine the side effect on hatchability. Acompletely randomized design
(CRD)was used toarrange the experimental units. Each treatment was
replicated four times, the data was taken 48 hours after treatment.
3.7.2. Larval stage:
Ten larvae (3rd Instar) in a petri-dishes containing filter papers were
sprayed to determine the mortality percentage. The each treatment was
replicated four times.
3.7.3. Adult stage:
Ten adult insects of C. propinque (Muls) in a petri-dish containing
filter papers were treated topically with the different treatments to study the
34
mortality percentage. Each treatment was replicated four times. The
treatments used in this experiments were as follows:
1.
25 g of NSKP per liter water.
2.
50 g of NSKP per liter water.
3.
75 g of NSKP per liter water.
4.
Actara® used at recommended rate of (1 gram bar 1.25 liter of
water).
5.
Control (water only).
3.7.4. Feeding of C. propinque (Muls) on aphids treated topically:
This experiment was carried out to determine the effect of treated
aphids on the feeding of the adult of C. propinque (Muls). The aphids
Aphis fabae (Scop) were sprayed with neem seed water extract (NSWE),
Actara and water (control). They were offered to adult insect of C.
propinque (Muls) to feed freely on them. The aphids were kept in Petridishes, and then the mortality percentage was calculated. Each treatment
was replicated four times.
The treatments were as follows:
1.
25 g NSKP per liter water.
2.
50 g NSLP per liter water.
3.
75 g NSKP per liter water.
4.
Actara® neonicotinoid used at recommended Rate (1gram bar 1.25
liter of water).
35
5.
Control (water only).
The number of live insects (Coccinellids) were counted in each dish
every 24 hours for three days.
Statistical analysis:
The collected data was statistically analyzed using ANOVA. The
means were separated by DMRT.
36
Eggs
Larvae
3
37
Adult
Plate 4. The test stages of C. propinque.
38
CHAPTER FOUR
RESULTS
Generally, the synthetic insecticide Actara® Nenicotionid active
ingredient thiomethoxam used with a recommended dose of (560g/ha) was
significantly better in reducing the number of Aphis faba (Scop) on faba
bean. The same insecticide resulted in a 100% mortality of Coccinellid C.
propinque (Muls). The test neem products proved to be less harmful to the
predator compared with Actara.
4.1.1. The effect of NSK Aqu-Ext, applied in the soil as drenching on
aphids (drenching experiment).
The result of this experiment illustrated that all treatments on the 1st,
2nd, 3rd, and 5th days after treatment showed highly significant difference
(P<0.01) when compared with the control. However, on the 4th, 6th, 7th days
there was no significant difference between NSK Aqu-Ext and control as
whereas there was a highly significant difference between Actara
insecticide and control.
The NSK Aqu-Ext (75g) gave high mortality on aphids (A. fabae
Scop) followed by (50g). There was no significantly different mortality
between the concentration (25g) and the untreated control (Table 1, and
Appendix A).
39
Table 1. Survival and corrected mortality (CM) of A. fabae (20 insects) fed on seedling of faba bean treated
with NSWE (soil application).
Days
1st
Treatments
2nd
3rd
4th
5th
6th
7th
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Control
89.43a
-
86.34a
-
89.43a
-
81.88a
-
89.43a
-
86.34a
-
86.34a
-
25g NSKP/l
80.49a
5
74.94b
8
78.79b
5
72.94a
5.19
86.34a
1.25
73.59a
3.80
75.90a
3.80
50g NSKP/l
65.19b
19
69.25b
8
61.72d
22
70.09a
13.90
77.70b
6.25
82.38a
2.03
77.41a
5.06
75g NSKP/l
45.06c
50
73.23b
8
69.82c
12
68.60a
13.24
89.43a
0.00
80.17a
2.28
86.34a
0.00
Actara®
0.57d
100
0.57c
100
0.57e
100
0.57b
100
0.57c
100
0.57b
100
0.57b
100
C.V%
SE±
12.53%
11.25%
7.27%
15.46%
7.16%
14.02%
11.73%
3.52
3.42
2.18
4.55
2.46
4.53
3.83
Figures followed by different letters are significantly different at P = 0.05 (DMRT).
40
4.1.2. The effect of NSK Aqu-Ext sprayed on the faba bean plants.
The statistical analysis of the results showed that there was highly
significant difference (P< 0.01) between all treatments and control on the
1st, 2nd , 3rd, and 4th days after treatment, however, on 5th, 6th, and 7th days
there was no significant difference between NSK Aqu-Ext and control
(Table 2, and Appendix B).
The three concentrations of neem seed water extract showed the same
effect on the first day after treatment. There was a significant difference
between the neem treatment and the synthetic insecticide Actara® which
gave a corrected mortality of 100%. No significant difference between the
control and all treatments was obtained on 5th, 6th, and 7th, days after
treatment indicating that this method of applying neem products was
ineffective.
4.1.3. The effect of NSK Aqu-Ext sparyed topically on the aphids.
Table 3and Appendix C summarized the effect of NSK Aqu-Ext,
applied topiclly on the faba bean aphids. A highly significant difference
between all treatments and the control was obtained on the 1st, 2nd, 3rd, and
4th days after treatment. However, there was no significant difference
between the NSK Aqu-Ext and control.
On the 1st day after treatment, all concentrations of NSK Aqu-Ext
gave the same results. The neem treatments were however, significantly
different from the control. On the 4th, 5th, 6th, and 7th, days after treatment,
all concentrations of neem seeds kernel extracts showed no significant
effect on aphid compared with the control.
41
Table 2. Survival and corrected mortality (CM) of A. fabae(20 insects) after foliar application of NSWE on
faba bean (post infestation).
Days
1st
Treatments
2nd
3rd
4th
5th
6th
7th
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Control
89.43a
-
89.43a
-
86.34a
-
89.43a
-
81.88a
-
81.88a
-
83.26a
-
25g NSKP/l
72.49b
12.5
65.55b
17.5
65.56b
16.46
78.46ab
7.5
86.34a
2.60
84.96a
1.3
80.17a
1.28
50g NSKP/l
74.77ab
12.5
57.72c
28.75
61.74b
21.52
68.02b
15
83.26a
1.30
84.96a
1.30
78.79a
2.56
75g NSKP/l
59.55b
26.25
70.76b
11.25
59.78b
25.32
68.02b
15
84.96a
1.30
77.41a
2.60
86.34a
1.28
0.57c
100
0.57d
100
0.57c
100
0.57c
100
0.57b
100
0.57b
100
0.57b
100
Actara®
C.V%
SE±
17.32%
6.58%
9.76%
12.53%
10.49%
11.97%
9.22%
5.14
1.87
2.67
3.82
3.54
3.95
3.03
Figures followed by different letters are significantly different at P = 0.05 (DMRT).
42
Table 3. Survival and corrected mortality (CM) of A. fabae( 20 insects) topically sprayed with NSWE.
Days
1st
Treatments
2nd
3rd
4th
5th
6th
7th
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Control
89.43a
-
89.43a
-
86.34a
-
83.26a
-
84.96a
-
80.49a
-
81.88a
-
25g NSKP/l
56.86b
30
84.96a
2.5
72.76b
10.13
80.17a
1.28
89.43a
2.56
83.26a
2.63
77.41a
2.30
50g NSKP/l
53.86b
35
69.82b
12.5
50.85c
39.24
83.26a
0.00
86.34a
1.28
86.34a
3.95
86.34a
2.60
75g NSKP/l
51.61b
38.75
61.10b
25
58.83c
27.85
86.34a
1.28
86.34a
1.28
86.34a
3.95
86.34a
2.60
Actara®
0.57c
100
0.57c
100
0.57d
100
0.57b
100
0.57b
100
0.57b
100
0.57b
100
C.V%
7.97
11.08
15.32
8.94
8.04
10.14
10.16
SE±
2.01
3.39
4.13
2.98
2.79
3.42
3.38
Figures followed by different letters are significantly different at P = 0.05 (DMRT).
43
4.2.1. The effect of NSK Aqu-Ext on the Egg hatchability of C.
propinque.
In this experiment all treatments resulted in highly significant
difference (P < 0.01), 48 hours after application when compared with the
control. The best hatchability was obtained in the treatment of 25g NSK,
followed by 50g and 75g respectively. The synthetic insecticide Actara®
gave zero hatchability in contrast to the control (Fig1and Appendix D).
4.2.2. The effect of NSK Aqu-Ext on the 3rd, larva instar.
There was no significant difference (P> 0.01) between the NSK (25g) ,
(50g) and the control on the 1st day after treatment. However, the
concentration (75g) gave a significantly different mortality compared with
the control. The same trend was observed on the 2nd day after treatment
concerning the effect of NSK–Aqu- Ext All neem treatments were
significantly different from the control. On the three days following the
treatment, Actara resulted in a significant different mortality of 100%
(Table 4 and Appendix F)
4.2.3. The effect of NSK Aqu-Ext on the Aduts of C. propinque (Muls).
Table 5 and Appendix E showed that there was a significant
difference (P<0.01) between all treatments and control. The mortality
caused by Actara was 100% on all days after the treatment. The NSK-AquExt concentration gave the same results on the 1st day after treatment and
the same trend was observed on the 2nd ,3rd, days after treatment.
44
.
90
80
Hatchability %
70
60
50
40
30
20
10
0
Control
25 gNSKE
50 gNSKE
75 gNSKE
Treatment
Fig. 1. The effect of topical application of NSK Aqu-Ext on the egg hatchability of
(C. vicina "Muls") 48 hours after treatment
45
Table 4. Survival and corrected mortality (CM) of C. propinque adults (10
insects) after direct spraying with neem seed water extract.
Days
1st
Treatments
2nd
3rd
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Control
89.43a
-
89.43a
-
84.96a
-
25g NSKP/l
67.50b
15
78.46ab
7.5
78.46ab
5.13
50g NSKP/l
59.41b
27.5
84.96a
2.5
65.47b
15.38
75g NSKP/l
67.94b
20
69.08b
17.5
71.97ab
10.26
Actara
0.57c
100
0.57c
100
0.57c
100
C.V%
14.38
14.51
15.16
SE±
4.10
4.68
4.57
Figures followed by different letters are significantly different at P = 0.05 (DMRT).
46
Table 5. Survival and corrected mortality (CM) of 3rd larvae (10 insects) of
C. propinque after direct spraying with neem seed water extract.
Days
1st
Treatments
2nd
3rd
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Control
82.93a
-
89.43a
-
84.96a
-
25g NSKP/l
70.25a
13.16
60.72b
30
84.96a
0.00
50g NSKP/l
75.30a
7.89
56.25b
35
78.32ab
10.26
75g NSKP/l
48.75b
42.11
66.77b
20
62.58b
20.51
Actara
0.57c
100
0.57c
100
0.57c
100
C.V%
SE±
22.46%
22.55%
18.39%
6.24
6.17
5.73
Figures followed by different letters are significantly different at P = 0.05 (DMRT).
47
4.2.4. The feeding of C. propinque (Muls) on aphids treated topically
with NSK Au- Ext.
On the frist day after treatment, there was no significant difference
(P> 0.01) between concentration 25g NSK Aqu-Ext and the control,
however, there was a highly significant difference among the other
treatments (Table 6 and Appendix G).
On the 3rd day after treatment, the results showed highly significant
difference (p<0.01) between 25g of NSK –Aqu- Ext,75g NSK Aqu-Ext
concentration and the control, but there was no significant difference
between the 50 g NSK Aqu- Ext concentration and control.
48
Table 6. Survival and corrected mortality (CM) of C. propinque adults (10
insects) fed on aphids treated topically with
neem seed water
extractNSWE.
Days
1st
Treatments
2nd
3rd
Surviving
C.M%
Surviving
C.M%
Surviving
C.M%
Control
89.43a
-
80.49a
-
76.03a
-
25g NSKP/l
76.43a
10
75.57ab
7.89
60.27b
18.92
50g NSKP/l
54.53b
35
74.00ab
5.26
74.00a
2.70
75g NSKP/l
55.39b
35
59.13b
23.68
64.33ab
13.51
0.57c
100
0.57c
100
0.57c
100
Actara
C.V%
SE±
18.59%
18.73%
14.91%
5.14
5.43
4.10
Figures followed by different letters are significantly different at P = 0.05 (DMRT).
49
CHAPTER FIVE
DISCUSSION
5.1. General:
In the last decade, the neem tree Azadirachta indica has come under
close scientific scrutiny as a natural source of noval pest control materials.
Neem derivatives were found to have systemic and contact effects against
many sucking and chewing insect pests. As Homopteran plant insect pests
generally feed by sucking plant sap, neem derivatives may affect them
systemically.
The natural enemies play a fundamental role in integrated pest
management programs (Michel et al., 1997) and the study of the side effect
of botanicals on them is very essential before being incorporated in any
control program. In the Sudan, Coccinellids are important predators in
several field crops specially in winter. A part from some limited laboratory
experiments, there is no extensive field research in Sudan to determine the
deleterious impact of neem products on natural enemies and other beneficial
insects (Osman and Bradley, 1993). Schmutterer, (1992) stated that only
little work has been generally carried our on the effects of neem on
parasitoids or predators.
5.2. The effects of NSK Aqu-Ext on Aphis fabae :
As shown in table 1. in the soil drenching experiment, the chemical
insecticide Actara® showed the best results in reducing the aphids numbers.
Mudathir (2000) found similar results, by using Sumicidin® in the field to
control aphids on okra. The effectiveness of NSK Aqu-Ext increased with
50
time up to the first three days after application and decreased in the
following last three days after treatment, this might be due to the high
concentrations of neem extracts in the plant tissue in first 3 days. Heyde et
al. (1984) found that the mortality of aphids increased up to (8 days) when
he used NSK powder. He went to say that the residual effect of bioactive
ingredients will be prolonged if a systemic effect (translocation of active
material) takes place, directing the uptake of material by the treated plant.
In foliar application – experiment, as shown in Table 2. Actara®
NSK Aqu-Ext were significantly effective in reducing aphids numbers
compared to the control, the Actara® gave highly mortality than NSK AquExt. This could be explained by the fact that neem products have no knockdown effect compared with synthetic pesticides. The NSWE treatment (Post
infestation) was significantly different (P < 0.01) on the first 4 days
following application. Lowery and Isman (1993) reported that neem seed
oil, 48 hrs following aphid infestation (post-infestation) were not
significantly different (P > 0.05) from the control. This insignificant effect
might be due to the slow absorption of neem oil by plants.
As shown in the Table 3, (the topical experiment), Actara® gave
highly mortality than NSK Aqu-Ext concentrations. Lowery et al. (1993)
found in a field experiment, that foliar applications of NSO resulted
significantly better control of M. persicae on pepper. The difference
between his results of the findings of present investigation could be due to
different preparations of neem and the different conditions under which the
experiments were undertaken.
51
5.3. The effects of NSK Aqu-Ext and Actara® on the different stages of
the Coccinellid Cheilomenes propinque (Muls):
As shown in Fig. 4, the Actara® pesticide gave zero hatchability
while the hatchability was slightly affected by NSWE. The same results was
obtained by other authors on different species of Coccinelld particularly
Coccinella septempunctata (Schmutterer, 1981 and Kaethner, 1990). The
two authors used AZT-VR-K (1000 ppm) and combined it with neem oil
(250 – 30000 ppm).
As shown in table 5, the insecticide Actara® spryed on Adults C.
propinque (Muls) topically gave 100% mortality. The same results were
obtained by (Sharma and Adlakha, 1981) who found that “Feniforthion”
was very toxic to other species of Coccinellid C. septempunctata whereas
“endosulfan” did not appear to be harmful. The effect of NSWE on adults is
harmless as compared with synthetic pesticide Actara®. When development
and growth are concerned, neem products can affect hormonal balance and
hence development and growth of young stages and later fertility of adults.
The same results were obtained by Hoelmer et al., (1990) who reported
mortality of the coccinellid beetles Delphastus pusillus and Scymnus sp
reared on leaf discs dipped in Morgosan-O and kept in sealed Petri-dishes
and glass vials for two weeks.
According to Schmutterer (1990) neem products have relatively weak
contact toxicity NSWE applied in the field at rate of 50g/l water proved to
be safe for the Coccinellid Scymnus sp. (Elshafie, 2001).
The topical treatment of (3rd instar) the larvae of Coccinellid C.
propinque (Muls), with neem concentrations of 25g and 50g gave no
52
significant difference (P < 0.01) on the first day after treatment compared
with the control. However, significant mortality occurred during the
following days.
Ossiewatsch (2000) reported the same results on the C.
septempunctata. He also reported that high mortality of larvae occurred in
laboratory when the insects were dipped in NSWE.
As shown by the results presented in Table 7 adults of Coccinellid
fed on aphid treated topically with NSWE, showed an increase in mortality
with increase concentrations of neem extract on the first day. These results
are supported by the findings of Hoelmer et al. (1990) who reported that
larvae and females of Coccinellids Delphastus pusills and Scymuns sp. fed
on eggs of Bemisia tabaci treated with Morgosan-O for several days
showed no mortality and females continued egg laying.
5.4. Conclusions and recommendations:

Neem seed kernel aqueous extract (NSKWE) represent a relatively
safe, and inexpensive, source of materials for integrated pest control
in
agroecosystem
exosystem
as
compared
with
synthetic
insecticides.

The application of neem into the soil could possibly decrease the
photochemical degradation and increase the possibility of
controlling some soil pests like nematodes.

The neem products applied systemically could be considered safe
for the natural enemies. However, the effect of neem on predators
53
and prasitoids of aphids should be studied more fully before any
practical application is carried out.

Neem seed kernel water extract proved to be less harmfull to the
aphidophagous Coccinellids compared with the chemical insecticide
Actara®. Therefore, it is recommended for use against agricultural
pests within the framework of IPM and organic farming.
54
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64
APPENDICES
65
APPENDIX (A)
The analysis of variance table of the effects of (NSKE) on aphid
(A fabae Scop) treated as soil drenching the plants.
ANOVA: Analysis of variance tables:
1st day:source
D.F
Sum of Squr
Mean squr
F. cal
F. table
Treatments
4
19975.931
4993.983
100.825
0.000
Error
15
742.968
49.531
Total
19
20718.90
C. V = 12.53%
SE= ± 3.52
2nd day:
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
18823.668
4705.917
100.431
0.000
Error
15
702.861
46.857
Total
19
19526.529
C.V. 11.25%
SE = ± 3.42
66
3rd - day:source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
19401.752
4850.438
254.024
0.000
Error
15
286.416
19.094
Total
19
19688.168
CV = 7.27%
SE = ± 2.18
4th day:
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
17386.103
4346.526
52.576
0.000
Error
15
1240.060
82.6761
Total
19
18626.163
C .v = 15.46%
SE = ± 4.55
5th –days
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
23573.682
5893.420
243.668
0.000
Error
15
362.794
24.186
Total
19
23936.475
C.V = 7.16%
SE = ± 2.46
67
6th – days:
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
20847.134
5211.748
63.563
0.000
Error
15
1229.913
18.994
Total
19
22077.047
C. V = 14.02%
SE = ± 4.53
7th – day:source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
21337.778
5334.444
90.897
0.000
Error
15
880.474
58.698
Total
19
22218.251
C. V = 11.73%
SE = ± 3.83
68
APPENDIX (B)
The analysis of variance tables at the effect of (NSKE) on the aphid
(A. fabae Scop) treated as foliar on the plants.
1st day :
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
19081.530
4770.382
45.119
0.000
Error
15
1585.928
105.729
Total
19
20667.458
C.V =17.32%
SE = ± 5.14
2nd- day:-
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
17996.050
4499.013
321.857
0.000
Error
15
209.674
13.978
Total
19
18205.725
C.V = 6.58%
SE = ± 1.87
3rd – day:-
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
16497.332
4124.333
144.178
0.000
Error
15
429.089
28. 606
Total
19
16926.421
C.V = 9.76%
SE = ± 2.67
69
4th –day :source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
19454.351
4863.588
83.498
0.000
Error
15
873.716
58.248
Total
19
20328.067
C.V = 12.53%
SE = ± 3.82
5th – day:-
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
22377.661
5594.415
111.894
0.000
Error
15
749.964
49.998
Total
19
23127.626
C.V =10.49%
SE = ± 3.54
6th day:
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
21529.498
5382.374
86.294
0.000
Error
15
935.587
62.372
Total
19
22465.085
C.V = 11.97%
SE = ± 3.95
70
7th day:
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
21426.886
5356.722
145.395
0.000
Error
15
552.637
36.842
Total
19
21979.523
C. V =9.22%
SE= ± 3.03
71
Appendix ( C)
The analysis of variance tables of the effects of (NSKE) on aphid
(A. fabae Scop) treated as topically on the aphid.
1st day:
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
16246.272
4061.568
251.366
0.000
Error
15
242.370
16.158
Total
19
16488.642
C.V = 7.97%
SE = ± 2.01
2nd day:-
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
20447.766
5111.941
111.196
0.000
Error
15
689.584
45.972
Total
19
21137.350
C.V = 11.08%
SE = ± 3.39
3rd day:
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
17144.346
4286.087
62.943
0.000
Error
15
1021.415
68.094
Total
19
18165.762
C. V = 15 .32%
SE = ± 4.13
72
4th day :source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
21954.051
5488.513
154.221
0.000
Error
15
533.829
53.589
Total
19
22487.880
C.V= 8.94%
SE = ± 2.93
5th day :source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
23819.896
5954.974
190.748
0.000
Error
15
468.286
31.219
Total
19
24288.183
C.V = 8.04%
SE = ± 2.79
6th day:-
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
22426.982
5606.746
120.034
0.000
Error
15
700.643
46.710
Total
19
23127.626
C. V = 10.14%
SE = ± 3.42
73
7th day:
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
21958.314
5489.578
120.146
0.000
Error
15
685.365
45.691
Total
19
22643.679
C.V = 10.16%
SE= ± 3.38
74
APPENDIX (D)
The analysis of variance of tables of the effects of (NSKE) on egg
hatchability of the C propinque (Muls) after 48 houre.
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
9407.170
2351.792
360.170
0.000
Error
15
975.305
65.020
Total
19
10382.475
C.V = 13.81%
SE = ± 4.03
75
APPENDIX (E)
The analysis of variance table of effects of (NSKE) on the adults
predation C propinque (Muls) treated as topically.
1st – day :source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
17887.197
4471.799
66.664
0.000
Error
15
1006.191
67.079
Total
19
18893.388
C.V. = 14.38%
SE = ± 4.10
2nd day:-
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
21372.545
5343.136
61.002
0.000
Error
15
1313.851
87.590
Total
19
22686.396
C.V = 14.51%
SE = ± 4.68
3rd day :-
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
18474.950
4668.738
55.873
0.000
Error
15
1253.387
83.559
Total
19
19928.337
C.V = 15.16%
SE = ± 4.57
76
APPENDIX (F)
The analysis of variance tables of the effect of(NSKE) on the larvae of
& ( 3rd in star ) of (C propinque Muls) treated or spraying topically.
1st day :
Source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
17699.286
4424.821
28.405
0.000
Error
15
2336.682
155.779
Total
19
20035.968
C. V = 22.46%
SE= ± 6.24
2nd day:
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
17282.065
4320.516
28.346
0.000
Error
15
2286.303
152.420
Total
19
19568.368
C.V= 22.55%
SE = ± 6.17
3rd day:
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
20378.270
5094.567
38.823
0.000
Error
15
1968.399
131.227
Total
19
22346.669
C.V = 18.39%
SE= ± 5.73
77
APPENDIX (G):
The analysis of variance tables of the effects all the adult predation
Coccinellids (C propinque Muls) with the aphids (A fabae Scop) treated
tropically with (NSKE).
1st day :source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
18430.089
4607.522
43.653
0.000
Error
15
1583.222
105.548
Total
19
20013.311
C.V = 18.59%
SE = ± 5.14
2nd day:-
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
17481.233
4370.308
37.090
0.000
Error
15
1767.464
117.831
Total
19
19248.697
C.V= 18.73%
SE= ± 5.43
3rd day:
source
D.F
Sum of Squr
Mean squ
F. cal
F. table
Treatments
4
15522.289
3886.572
57.597
0.000
Error
15
1010.617
67.374
Total
19
16532.906
C.V = 14.91%
SE = ± 4.1
78
79