Advances in artificial diets for mass rearing of natural enemies

ACHARYA N.G RANGAAGRICULTURAL UNIVERSITY
Agricultural college, Bapatla
Course No : ENT 507
Course Title : Biological control of crop pests and weeds
Topic : Advances in artificial diets for mass
rearing of natural enemies
Submitted to,
Dr. D.V.Sairam kumar
Professor
Department of entomology
Agricultural college
Bapatla.
Submitted by,
P. Vijay babu
BAM 18- 41.

What is an artificial diet?
 An artificial diet is defined as any diet that is not
the natural food of an insect.
 It may be composed of,
 Plant extracts
 Ground plant material
 Semi-purified proteins
 Lipids
 Polysaccharides.
 Combined with other nutrients and water, or it
may be composed entirely of pure chemicals.
3

Why it was developed?
1) Rearing of insects on their natural host is time
consuming due to regular change of plant material.
2) Involves more man power.
3) Lot of plant material is wasted.
4) Frequent handling of culture results in mortality of
insects.
5) Insects can not be reared round the year.
4

Types of diet:
 There are three basic groups of artificial diets
with respect to the components they are,
1. Oligidic
2. Meridic
3. Holidic
5

 OLIGIDIC:-
 Crude natural materials + liver or other animal
products.
 E.g: Casein & yeast for a Flour beetle and yeast,
cholesterol & water for larval wax moth.
 MERIDIC:-
 Defined chemical + one or more ill defined
components such as casein.
 These diets may also be highly complex.
6

 HOLIDIC:
 Entirely of known pure chemicals.
 Most desirable for critical nutritional studies.
 Most artificial diets used for insect rearing are of
meridic type .
 Several insects and entomophagous insects can now be
conveniently reared on such artificial diets (Singh et
al,1982).
7

Advanced artificial diets :-
1. Rearing of Mallada basalis (Neuroptera:Chrysopidae)
on modified artificial diets.
2. Improving the artificial diet for adult of seven spotted
ladybird beetle, Coccinella septumpunctata.
3. In Vitro Rearing of Trichogramma minutum Riley
(Hymenoptera:Trichogrammatidae) for Ten
Generations, with Quality Assessment Comparisons of
in Vitro and in Vivo Reared Adults.
4. Optimization an Optimal Artificial Diet for the Predatory
Bug Orius sauteri (Hemiptera: Anthocoridae).
8

1. Rearing of Mallada basalis
(Neuroptera:Chrysopidae) on modified
artificial diets:-
 Mallada basalis (Walker) has the
potential to be a valuable biological
control agent because of its,
1. Predatory abilities
2. Strong reproductive capacity
3. Broad prey range.
 The larvae of this lacewing are
general predators, whereas the
adults feed on nectar, honeydew,
and pollen.
9

 Previous investigations suggest its potential as a
biological control agent against several species of pests
including,
 Phyllocnistis citrella
 Aphis spp.,
 Nipaecoccus filamentosus (Cockerell),
 Diaphorina citri
 Panonychus citri on citrus;
 Aphis gossypii (Glover) on sweet pepper;
 Tetranychus urticae
 Tetranychus kanzawai on strawberry;
 Planococcus citri on Indian jujube; and
 Icerya aegyptiaca.
10

Mallada spp. Feeding on citrus and
maize aphids.
11

 This study aimed to improve on a previously used
artificial diet for M. basalis,
 To achieve a longer oviposition period and
 Greater survival rate and fecundity.
 Analyzed the development, survival, longevity, and
reproduction of M. basalis (F1 and F2 generations) fed
two artificial diets (AD1 and AD2).
 Both diets contained chicken egg yolk, beer yeast
powder, honey, trehalose, seawater spirulina, and
potassium sorbate.
12

 AD1 also contained sucrose and vitamin C.
 It is found that diet AD1 supported development and
reproduction better than AD2 and the diets in our
previous study.
 These findings may contribute to the mass rearing of this
economically important predatory green lacewing.
13

Preparation of artificial diets:-
 The artificial diets were formulated based on that
described by Ye et al 2000.
 Two artificial diets (AD1 and AD2) were prepared for
M. basalis larvae, and their composition, as well as the
composition of the previous artificial diet (AD0).
 Compared with previous diets, seawater spirulina
was added to both AD1 and AD2 as a dietary protein
source. (55 and 70% protein, more than beef, chicken,
and soybeans)
14

 In addition, sucrose and vitamin C were removed from
AD2 to determine whether they were essential
ingredients.
 If not essential, their subsequent removal from the diet
would reduce food costs.
 Diet ingredients were blended in a food processor for 3±5
min until the entire mixture was homogenous.
 A 10 ×10 mm Parafilm membrane was stretched to about
3x its original length and width.
17

 The diet was placed centrally on the membrane which
was folded and stuck tightly together.
 The artificial diets were prepared every 2 weeks and kept
in a refrigerator at 5˚C.
 The weight of each diet packet was 0.05 g, while the
weight of the diet itself was 0.03 g.
18

Conclusions:-
 We found that Mallada basalis was able to develop and
reproduce when fed artificial diets AD1 and AD2.
 However, the F1 adult longevity and F2 oviposition period
in the AD1 treatment was significantly longer than that in
the AD2 treatment.
 Results indicated that the offspring of M. basalis fed AD2
were less thrifty (using money and other resources
carefully and not wastefully).
19

 Only diet AD1 contained sucrose and Vitamin C, which
suggests that these ingredients may be important nutrient
sources leading to the superiority of AD1.
 The duration of larval development of M. basalis reared
on AD1 was longer than that previously found when
reared on C. cephalonica eggs.
 M. basalis fed on AD1 produced more eggs than when
fed on the four natural prey species.
20

 The developmental duration of F1 immature M. basalis
reared on AD1 was longer than that reared on an artificial
diet consisting of the same ingredients but with no
seawater spirulina.
 A high content of the dietary alga is suggested to support
good larval growth and reduce developmental time.
21

2. Improving the artificial diet for adult of
seven spotted ladybird beetle,
Coccinella septumpunctata.
(Y. cheng, J.R. Zhi, F.L.Li, W.H. Li and Y.H. Zhou)
First published online 19th september, 2017.
22

Introduction:-
 The lady bird beetle, Coccinella septumpuntata is
considered to be an important biocontrol agent for
soft bodied insects like,
 Aphids
 Whiteflies
 Jassids and
 Small lepidopterous larvae (Victor,1997)
23

 It is a good biocontrol agent for
thrips and whiteflies in
greenhouse crops
(Deligeogidis et al. 2005).
 Both Coccinella septumpucntata
adults and larvae are primarily
predators of aphids (Rizvi et al.,
1994)
 However, due to impact of climate
and farming operations, it is often
difficult to maintain sufficient
levels of natural supply of the
beetles to control pests effectively
(sun & Wan 1999; Zhang et al.,
2014;).
24

 Therefore, it is necessary to artificially rear them as
supplement predator for effective field control of pests.
 Currently, lady bird beetles are reared mainly using
aphids as food.
 This method become costly & time consuming for the
mass production of the insect (Ashraf et al., 2010; Bonte
et al., 2010).
 Therefore, successful & continuous mass rearing of
beetles at a commercial level is important for this
purpose.
 Hence, it requires the availability of artificial diets.
25

Earlier works :-
 Studies on artificial diets for rearing of ladybird beetles
were initiated in the 1950’s.
 Smirnoff (1958) reported a prey powder based artificial
diet that successfully reared a variety of predatory
ladybird beetles.
 Later on, non-prey compositions and non-insect source
components were studied for the diet for various species
of ladybird beetles (Singhinolfi et al., 2008; Zhang et al.,
2011; Tan et al., 2015).
26

Materials and methods :-
 An orthogonal array design with 16 factors at 2 levels
was performed to develop an artificial diet.
 The parameters of,
 Weight gain
 Survival rate of adults
 Preoviposition period
 Fecundity
 Hatching rate; of diet fed adults were monitored.
27

 The 16 factors included here are,
1. Pork liver
2. Infant formula
3. Sucrose
4. Olive oil
5. Yolk
6. Corn oil
7. Yeast powder
8. Cholestrol
9. Casien
10. Casien hydrolysate
11. Vitamin powder
28
Cont…..

12. 65% juvenile hormone ΙΙΙ
13. Protein powder
14. Vitamin E
15. Honey
16. Pumpkin.
29

Preparation of artificial diet:-
1. Pork liver and pumpkin were purchased from local
market.
2. The pork liver was grinded into a pasta with a tissue
titurator.
3. Peeled pumpkin was steamed & then grinded as well
4. The ingredients were weighed and mixed.
30

5. Then, added to 600mL sterile water containing 12g
molten agar @ 40-50oc.
6. Later, blended for 3 min to generate a homogenous &
semi-solid diet mixture.
7. The mixture was cooled to room temperature and
stored in refrigerator at a 4oc.
31

Feeding with artificial diets & aphids:-
1. The newly emerged adults were sexed and paired, 50
pairs for a treatment were obtained within a single day.
2. In both the orthogonal array design trail and follow-up
validation experiment comparing the optimized diet
against an aphid diet.
3. Each pair was placed in a 500mL plastic jar where the
semi-solid diet was supplied on a small piece of plastic
strip at the bottom.
32

4. The pair was provided daily with fresh artificial diet,
absorbent cotton with distilled water & shrinking paper.
5. Egg masses laid on paper were collected 3 times a day.
6. Transferred to a filter paper moistened with distilled
water in a 9cm Petri dish.
7. For feeding with Aphis craccivora, freshly collected
aphids(roughly 500) were added to each jar daily
through the nylon netting(lid).
33

Results & discussion :-
Results of artificial diets feeding are shown in the table,
1. Ten days after feeding, the T16 gave the greatest weight
gain i.e.,11.83mg.
2. T12 generated the lowest weight gain with 2.47mg per
insect.
3. The 50 day survival rate was the highest 81.67% in the
treatment T13 & lowest(41.63%) with T4.
4. Eggs were produced in all the treatments.
5. But, T6 has the shortest pre-oviposition period (6.4
days) & longest for T5 (13.1 days).
35

6. T9 generated the highest number of eggs per female
(284.4 eggs), while T2 was the lowest (117.2 eggs).
7. The T9 gave the highest hatching rate (81.36%), while
the T2 was the lowest (58.38%).
36

8. Study revealed that,
1. Yeast powder and honey – improved weight gain.
2. Pork liver & sucrose - affecting survival rate.
3. Juvenile hormone & pumpkin – affecting pre
oviposition period
4. JH+ sucrose+ pork liver - impact on fecundity
5. Yeast powder+ yolk - hatching rate of adult.
9. Pork liver is main component of artificial diet.
10.As it is rich in protein, cholesterol & minerals.
11.Pumpkin shortened the pre oviposition rate of insect.
12.Addition of vitamin E promoted reproduction of
beetle.
38

Conclusion :-
 The diet developed is suitable for rearing
Coccinella septumpunctata.
 The diet fed adults improved survival rate, fecundity,
shortened preoviposition period compared to previous
reports (Sun & Wan, 1999; Singhinolfi et al., 2008;
Asharaf et al., 2010; Lu et al., 2015).
 Previous diets contained of natural preys like, aphids,
pupae of wasps or eggs of Corcyra cephalonica (Rojas
et al., 2016)
39

 But the diet here developed is free from such preys which
saves a lot of time for maintaining these preys.
 The cooking process of diet is relatively simpler & quick.
40

3. In Vitro Rearing of Trichogramma minutum
Riley (Hymenoptera:Trichogrammatidae) for
Ten Generations, with Quality Assessment
Comparisons of in Vitro and in Vivo Reared
Adults
41

INTRODUCTION:-
 Egg parasitoids in the genus Trichogramma are widely
used for augmentative biological control programs for
insect pests.
 Considerable work has been done on the development of
artificial diets for Trichogramma spp.
 Many of these efforts have involved diets containing
hemolymph (Guan et al., 1978; Hoffman et al., 1975).
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 Development of artificial diets for Trichogramma would
foster the implementation of automated mass rearing
systems.
 The development of artificial diets has the potential to
revolutionize the way we produce these insects for
biological control, by permitting the introduction of
automated mass rearing systems.
43

Automated mass rearing system
44

Automated mass rearing system 45

Key Points:-
 Trichogramma minutum Riley were reared for 10
generations on an artificial diet containing,
 Yeast extract,
 Free Amine III,
 Non-fat dry milk,
 Chicken egg yolk,
 Chicken embryo extract and
 Manduca sexta (L.) egg liquid.
46

Non-fat dry milk Chicken embryo extract
48

Chicken egg yolk extraction process
49

Eggs of Manduca sexta from which liquid is extracted
50

 Quality control parameters like,
 Adult longevity,
 Sex ratio,
 Pupation rate,
 Percentage of pupae to emerge as adults,
 Adult female body length,
 Number of Helicoverpa zea (Boddie) eggs parasitized
by a female
 Percentage of deformed females
 Were assessed and compared to insects reared in
vivo on irradiated H. zea eggs.
51

 The diet used in these studies consisted of six
components, including one insect-derived component.
 The 10% suspension of nonfat dry milk was prepared
and heated in a boiling water bath (100°C) for 15 min.
 The chicken egg yolk was obtained by gently opening
one end of an egg, which had been surface sterilized with
70% ethanol for 10 min, and withdrawing the yolk with a
sterile syringe.
Artificial Diet:-
52

 The chicken embryo extract was obtained by
homogenizing embryos (incubated at 39°C for 10 days)
with saline (1:1).
 To remove the accessory gland secretion from the
exterior surface of the eggs, the eggs were mixed with
Alconox .
 The mixture was then manually ‘‘scrubbed’’ together in
cheesecloth and then rinsed several times in water.
 To prevent melanization of the M. sexta egg liquid, the
eggs were heated in a 60°C water bath for 10 min.
53

• Many Alconox products have been
used for decades and are widely
accepted throughout
the medical device industry
for device cleaning during and
after device validation.
54

 The M. sexta eggs were then surface sterilized in an
aqueous solution of 0.1% Clorox (The Clorox Co.,
Oakland, CA), rinsed four times, with sterile water, and
air dried in a laminar flow hood at 24± 1°C.
 Finally, the eggs were homogenized, using a Virtishear
homogenizer.
 The homogenate was filtered through two layers of sterile
cheesecloth to remove particles of egg chorion.
55

 The artificial diet also contained 5% Free Amine III which
has been shown to stimulate oviposition by T. minutum
females.
 The prepared diet had a pH of 6.6 and an osmolarity of
280 mOsm kg2/L.
 Before use for in vitro rearing, the diet was assayed to
insure that T. minutum eggs would hatch in the diet and
that there was no fungal contamination.
57

Wax Artificial Eggs (WAEs) :-
 Wax artificial eggs (WAEs), which consisted of liquid
droplets of diet coated with a thin layer of a paraffin-
Vaseline mixture were used in these experiments to
induce T. minutum females to oviposit.
 WAEs were prepared using a mixture of paraffin (75%)
and Vaseline (25%) heated to 65°C in a glass tube, which
was held in a water bath (Hagen and Tassen, 1965).
59

 Artificial diet, which partitioned from the paraffin–
Vaseline mixture to form the bottom layer, was added to
the tube.
 A micropipette (100 μl) was inserted into the tube (to the
bottom), withdrawn, and touched to a microscope slide.
 The paraffin–Vaseline mixture formed a thin layer on the
surface of the artificial diet, forming a droplet 2–3 mm in
diameter, a WAE, on the slide.
60

 A microscope slide, with 60 WAEs attached, was
exposed to 1-day-old adult Trichogramma (10–20
females per WAE) taped together with masking tape
for 6 hrs, at room temperature (24±1°C).
 Four slides, with a total of 240 WAEs, were used for each
generation in the in vitro rearing, which resulted in the
production of thousands of Trichogramma adults each
generation.
62

WAEs placed on a microscopic slide (which are exposed
for oviposition by Trichogramma minutum). 63

Trichogramma minutum laying eggs on WAEs
64

In Vitro Rearing:-
 WAEs, which had been exposed to parasitoids, were
surface sterilized in a 75% alcohol solution for 6 min
followed by several rinsings in sterile reverse osmosis
water.
 The excess water was removed with a sterile filter paper.
 The diet and T. minutum eggs from each WAE were
transferred to a well in a Falcon 3070 Micro test III (96-
well tissue culture plate), using a Pasteur pipette, which
had been stretched to a very fine tip.
65

Falcon 3070 Micro test III
(Culture plate) Pasteur pipette
66

 The process was carried out aseptically in a laminar flow
hood.
 The tissue culture plate was covered and sealed with
masking tape and placed in a 250-mm-ID heavy glass
desiccators which contained 100 ml of water, to maintain
ca. 100% RH.
 For larval development, the desiccator was held in the
laboratory at 24 ± 1°C under a 14L:10D photoregime.
67

 During the last larval instar the micro-titer plate was
opened, cleaned if any excess diet was present, covered,
and then held at 75% RH in a dessicator with 300 ml of a
saturated sodium chloride solution (Wylie, 1965) until
adults emerged.
 The adults produced on artificial diet were used for the
next generation of in vitro rearing and for quality control
assessments.
69

Trichogramma minutum pupae grown in the WAEs
70

Conclusion:-
 The development time was longer for in vitro reared
insects, but there were more deformed females in the in
vivo culture.
 The sex ratio, however, was generally not significantly
different between the in vitro and in vivo cultures.
 The in vitro reared females generally were larger, lived
longer, and parasitized more H. zea eggs.
71

4. Optimization an Optimal Artificial
Diet for the Predatory Bug Orius
sauteri (Hemiptera: Anthocoridae)
72

Introduction:-
 The flower bug Orius sauteri is
an important polyphagous
predator that is widely used
for the biological control of
mites and aphids.
 This study investigated the
optimal ingredients of an
artificial diet for raising
O. sauteri using a
microencapsulation
technique.
Orius sauteri, preying on
Thrips palmi.
73

 The ingredients included
 Egg yolk (vitellus),
 Whole-pupa homogenate of the Tussah silk moth
(Antheraea paphia),
 Honey,
 Sucrose,
 Rapeseed (Brassica napus) pollen and
 Sinkaline (choline).
 Tested 25 combinations of the above ingredients using
an orthogonal experimental design.
74

 A microencapsulated artificial diet overcame many of the
difficulties highlighted by previous studies on the mass
rearing of O. sauteri.
 Optimization of the microencapsulated artificial diet
directly increased the biological and physiological
characters investigated.
 The use of artificial diets to rear biological control
arthropod agents has been studied for over 30 years.
75

 As with other predacious Orius spp. (e.g. O. tristicolor, O.
laevigatus and O. albidipennis), introducing natural prey
as the primary diet was the initial method used for
artificial rearing of O. sauteri.
 However, a natural prey diet was unable to support the
O. sauteri population throughout the year because of
environmental condition limitations and the need to rear
prey.
76

 Therefore, successful continuous mass rearing of O.
sauteri, particularly at a commercial level, required the
development of alternative artificial diets, which were
initially based on the artificial rearing of other Orius spp.
 In addition to the components of the diet, its state (liquid,
solid, etc.) can also influence rearing success, given that
Orius species have sucking mouth parts only.
 Although a promoted liquid diet could be easily ingested
by sucking in Orius similis, it led to high insect mortality
because the liquid feed was sticky.
77

 However, a solid artificial diet is not suitable for insects
that feed using sucking mouthparts.
 In addition, a solid diet is also unsuitable for rearing
Orius spp. because it can easily become contaminated
and dried out.
 To overcome these issues, microencapsulation was
introduced !
 Which is an advanced packaging technique in
widespread use for packaging microbial agents and
chemical or food products in the form of microcapsules
to promote the quality of artificial diets.
78

Microencapsulation Technologies:-
 Process of coating small particles of solids, liquids, or
gaseous components, with protective coating material.
 Size ranged from 2-5000μm.
 The microencapsulation process can be applied for a
various purpose such as,
79

1) To protect the core material from degradation and to
reduce the evaporation rate of the core material to the
surrounding environment.
2) To modify the nature of the original material for easier
handling;
3) To release the core material slowly over time at the
constant rate
4) To prevent unwanted flavor or taste of the core
material;
5) To separate the components of the mixture that would
react one another.
80

Overview:-
 The material that is encapsulated is called as core
material, the active agent, internal phase, or payload
phase.
 The substance or material that is encapsulating the core
is called as wall material or coating material.
 Three types of encapsulates are as follows,
 Reservoir type
 In matrix type
 Coated matrix
82

 In reservoir type, the active agents form a core
surrounded by an inert barrier (Gelatin and pectin).
 In matrix type, the active agent is dispersed or dissolved
in an inert polymer.
 Coated matrix type is a combination of first two.
83

Stock colony:-
 Healthy Orius sauteri were selected from the field
collection and 25 males and females were reared in a
custom-made cage.
 The reproduction of these experimental populations
depended on a daily supply of
 Tetranychus cinnabarinus mites as prey
 Fresh white kidney bean Phaseolus vulgaris seedlings as
an ovipositioning substrate.
84

Optimization of artificial diets for O. sauteri
rearing:-
 Artificial diet ingredients and orthogonal
collocation:-
 An orthogonal test design was used to investigate the
optimal artificial recipes according to the
developmental and reproductive performances.
 The optimization experiments were performed using
six ingredients and five concentration gradients, as
follows:
85

 Egg yolk--------------(0, 10, 20, 30, 40 g),
 Tussah pupae
Lyophilized powder-(10, 20, 30, 40, 50 g),
 Sucrose ---------------( 0, 5, 10, 15, 20 g),
 Honey -----------------( 0, 5, 10, 15, 20 g),
 Rape pollen----------(0, 5, 10, 15, 20 g)
 Sinkaline --------------(0, 5, 10, 15, 20 g)
(choline chloride)
 In total, 25 types of artificial diet were investigated.
86

Production of artificial diet
microcapsules:-
 The raw materials were dissolved in distilled water in a
clean 300mL glass beaker using the concentrations.
 The final constant volume was filled to 500mL in a
volumetric flask.
 The mixed artificial diet solutions from No. 1 to No. 25
were used as the core material in the
microencapsulation process.
88

 A general complex coacervation method was used to
make the artificial diet microcapsules (ADMs).
 The optimal basic chemical materials for artificial diet
microencapsulation for feeding O. sauteri to be as
follows:
1. Sodium alginate 2%,
2. Chitosan 0.6%
3. A 13:1 ratio of core material to wall-forming material
89

 Production of the ADMs involved eight steps:
1. Raw material treatment
2. Sterilization
3. Proportioning
4. Emulsification
5. Granulation
6. Rinsing
7. Filtration
8. Packaging .
90

The ADMs were made as follows:-
 A 2% mass concentration calcium chloride solution was
prepared using distilled water.
 A prepared artificial diet solution and quantificational
sodium alginate were mixed using a magnetic stirrer for
5 min at 3000 rpm to ensure uniform mixing.
 The mixture was then added to a Top-5300 model
medical micro-injection pump.
91

 We adjusted the pinhead of the injection pump to almost
5.0 cm to the surface of the calcium chloride solution.
 After turning on the power, the injection pump dropped
the mixture into the calcium chloride solution under 5
bars.
 The dropping speed could be controlled by adjusting the
pressure of the compressed air.
92

 The ADMs prepared were maintained in calcium chloride
solution for at least 5 min to establish their stability, and
were then moved to a quantificational chitosan solution
and shocked regularly for 13 min.
 This resulted in calcium alginate-chitosan-sodium
alginate colloid particles that were kept in 0.15% sodium
alginate solution for 30 min.
 Then, put into a 0.055 mol/L sodium citrate solution for
10 min.
93

 After being rinsed in physiological saline, the
microcapsules were also gently washed with distilled
water five times.
 Finally, the ADMs were conserved in airproof plastic
bags and stored in refrigerator until required.
94

Bioassay of O. sauteri fed on optimal ADMS :-
 Based on the data from the above experiment, we
produced ADMs containing the four recipes that had
resulted in optimal performance of selected biological
traits.
 Total nymphal development duration (ADM-D),
 Survival ratio (ADM-S),
 Oviposition duration (ADM-O)
 Female fertility (ADM-F)
96
• which were then offered to adult female O. sauteri.
• Orius sauteri female adults reared with T. cinnabarinus
were used as controls.

Population dispersal in greenhouses:-
 In total, 200 optimal ADM-D diet-fed newly emerged
O. sauteri female adults were maintained in a
transparent plastic box (15×20×20 cm) with a daily
supply of abundant ADMs.
 The comparison of O. sauteri dispersal rates was
conducted in a glass solar green house
 2000 tomato Lycopersicon esculentum var. cerasiforme
plants in an experimental area of 180 m2 (3665 m).
97

 The prey, leaf mites T. cinnabarinus, were added to the
plants at a density of 15 mites per 30 cm2 leaf area in
each experimental section.
 The plastic box was placed at the release site and the
cover removed.
 Ten tomato plants were selected random on which to
check the number of O. sauteri in each experimental
section at 24 h, 48, and 72 h.
98

 Also observed the population dispersal rate of O. sauteri
fed on the other 3 optimal ADMs, and on T. cinnabarinus
(control group).
 Each optimal ADM-reared treatment and control group
was replicated ten times.
 SPSS 16.0 software was used for the statistical analysis.
100

Results:-
 In total, 25 groups of ADMs were produced to feed
O. sauteri and the development duration from newly
hatched first instar nymphs to adult eclosion was
recorded
 With the exception of recipe No. 24, all the artificial diets
increased the total developmental duration compared
with the control.
 We repeated the survival measurements by feeding
nymphs with the optimum ADM, which resulted in a
90.7% survival rate
101

 The oviposition duration of insects fed the optimal ADM
was 7.83 d.
 Found that the natural organic components (egg yolk
and honey) had more positive effects on development
and female fertility,
 Pure artificial diet without any insect components may
cause many regression of reared Arthropod In
development, eclosion, oviposition and fertility.
102

Conclusion:-
1. The use of a microencapsulated artificial diet overcame
many of the negative effects recorded by previous
studies on the use of artificial diets for the mass rearing
of O. sauteri in terms of,
 Having a uniform shape
 Liquid diet stabilization and so on.
2. The introduction of sinkaline and honey was beneficial
in terms of their effects on the biological and
physiological characters examined, compared with
traditional artificial diet recipes.
103

4. There has been a report showing that provision of
pollen to O. laevigatus under nutrient limited conditions
can avoid cannibalism.
5. Sucrose is an efficient component in many artificial
diets for the mass rearing of arthropods, where it is
used as a feeding stimulant.
6. The determination of optimal ADMs could help increase
the effectiveness of biological control on the basis of
different requirements for artificial reproduction or field
release.
104

7. Our results suggest ADM-O and ADM-F could be used
for the mass rearing of Orius for use in inoculative
biological control systems, because of
high reproductive & copulatory efficiencies.
8. A low efficiency of mating preference of O. sauteri
reared on ADM-D or ADM-S was recorded
9. Such insects might be more beneficial for inundative
augmentative biological control because of a short
period of development, that could permit simultaneous
mass release.
105

References:-
 Ashfaq, M.N. Abida andGulam, M.C. 2002. A new technique for
mass rearing of green lacewing on commercial scale. Pakistan
Journal of Applied Sciences 2(9): 925-926.
 Arijs, Y., And P. De Clercq. 2004. Liver-based artificial diets for the
production of Orius laevigatus. Biocontrol 49: 505-516.
 Bonte, M., De Clercq, P. 2008. Developmental and Reproductive
Fitness of Orius laevigatus (Hemiptera: Anthocoridae) Reared on
Factitious and Artificial Diets. J. Econ. Entomol., 101: 1127-1133.
 Bonte M., De Clercq, P. 2010. Influence of predator density, diet
and living substrate on developmental fitness of Orius laevigatus.
J. Appl. Entomol., 135: 343–350.
 Cohen, A. C. 2004. Insect Diets Science and Technology. CRC Press,
New York. 324 pp. FERKOVICH, S. M., AND J. P. SHAPIRO. 2007.
Improved fecundity in the predator Orius insidiosus (Hemiptera:
Anthocoridae) with a partially purified nutritional factor from an
insect cell line. Florida Entomol. 90: 321-326.
106

 Cohen, AC. 2004. Insect diets: Science and technology. Cohen AC
(ed), CRC Press, Boca Raton, FL. 324p.
 Cônsoli, F.L., Grenier, S. 2010. In vitro rearing of egg parasitoids,
in “Progress in Biological Control- Egg parasitoids, In: Cônsoli
FL., Parra JRP., Zucchi RA. [eds], Agroecosystems with emphasis
on Trichogramma”, Springer Dordrecht, Chap. 11, pp. 293-313.
 Durmus, Y., Büyükgüzel, E., Terzi, B., Tunaz, H., Stanley D.,
Büyükgüzel, K. 2008. Eicosanoids mediate melanotic
nodulation reactions to viral infection in larvae of the parasitic
wasp, Pimpla turionellae. J. Insect Physiol., 54: 17-24.
 Grenier,S. 2011. Artificial rearing of entomophagous insects,
General outlines. Proceedings of Inter. Sympo. Mass Production
and Commercialization of Arthropod Biological Control Agents
(ISMC), Beijing, China, p.4-5.
 Grenier, S. 2009. In vitro rearing of entomophagous insects - Past
and future trends: a minireview. Bull. Insectology, 62: 1-6.
107

 Jiang L, Han SC, Li ZG, Lu X, Guo Q, Ye JW. Life table of an
experimental population of Mallada basalis and population
construction in the field. Forest Pests and Diseases. 2013; 32(4):
13±15, 23.
 Ribeiro, LJ. and Freitas, S.D. 2000. Influence of food on Chrysoperla
externa (Hagen) reproductive potential. Revista-de-Agricultura-
Piracicaba 75 (2): 187-196.
 Ye JW, Dai JQ, Li J, Li ZG, Lu YY, Han SC, et al. Development and
reproduction of Mallada basalis (Neuroptera: Chrysopidae) on
artificial diet. Florida Entomologist. 2015; 98(4):1072±1076.
108

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