AW-IPM.pdf

AW-IPM : Development and Field Applications
Siddu Lakshmi Prasanna
Ph.D. Scholar
Department of Plant Pathology
1

SIDDU LAKSHMI PRASANNA 2
Introduction
• Pest control and pest management
• Why we need to stop injudicious use of pesticides?
• Need of IPM
• Constrains in IPM implementations
• Difference between different types of Pest Management
Area Wide Pest Management
• How AWIPM differs from IPM?
• History of AWIPM programmes across the world
• Status or history AWIPM programmes in India
• Approaches of AW-IPM
• Models to be followed in AW-IPM
Case histories of AW-IPM programmes
• Benefits of AWIPM programmes
• Conclusions
Contents

Integrated Pest Management (IPM):
• Integrated pest management (IPM)
combines the use of biological,
cultural, mechanical and chemical
practices to control insect pests in
agricultural production.
• It seeks to use natural predators or
parasites to control pests, using
selective pesticides for backup only
when pests are unable to be
controlled by natural means.
• The four work hand in hand to
provide targeted, effective, long-
term pest management, and each
category plays a special role.

bio-physical
communication
personal
socio-
economic
technological
Constraints experience
d by IPM and Non-
IPM farmers
Constraints of IPM

Conventional Pest Management
• Chemical intensive
• Largely reactive to pest outbreaks
• Less emphasis on prevention
• Emphasizes killing pests directly
• Major purpose of site visits is to apply
pesticides
• General and widespread use of pesticides
Integrated Pest Management
• Knowledge intensive
• Systematic program of long term pest control
• Major emphasis on prevention of pest
problems
• Emphasizes modification of conditions that
favor pests
• Major purpose of site visits is to inspect and
monitor
• Pesticide use is to limit in terms of types,
amount and locations
• It is a field approach
Vs

An area-wide insect control programme is a long-term planned campaign against a pest insect population in a
relatively large predefined area with the objective of reducing the insect population to a non-economic status
(Lindquist, 2001).
• AWM is particularly relevant for pest species that are mobile, have a wide host range (crop and non-crop), and are
locally generated in the farming system, and enables management strategies on a larger-scale that may be more
effective than a paddock-by-paddock approach.
• AWM has two key objectives:
1. reduce overall pest pressure in participating regions by manipulating the size of the local population
2. manage insecticide resistance through coordinated rotation of insecticide groups.
Area wide- Integrated Pest Management

Area Wide Pest Management
1. Its treats all habitats of Pest infestation
2. It is implemented by an organization solely
dedicated to pest management in a region
3. It is a multiyear planning approach and
proactive in nature
4. It relies on both traditional and advanced
tactics of Pest management
Conventional approaches
1. It Defend only valuable entities like crop,
livestock from direct pest attack
2. It is Implemented by individual producers
3. It requires minimal forward planning and
reactive in approach
4. It relies on traditional tactics of pest
management
(Lindquist, 2000)

• not in abandoned crops, alternate hosts, backyard
hosts or on wild hosts.
• significant untreated refugia of the pest remain from
which recruits re-establish damaging densities of the
pest population.
• including abandoned crops, alternate hosts,
backyard hosts or on wild hosts.
• no significant untreated refugia of the pest remain
from which recruits can re-establish damaging
densities of the pest population.

Approaches of AW-IPM
Sterile
insect
technique
Mating
disruption
with
pheromones
Genome
editing
through
CRISPR
Enterobacter
(Gut
microbiome
of insects)
Cytoplasmic
incompatibility by
Wolbachia
release of insect
carrying
dominant lethal
(RIDL)
Male
Annihilation
Technique
Inundative
release of
parasitoid
insects
(Biological
control)

Sterile Insect Technique (SIT)
SIT defined as ―A method of pest control using area wide inundative releases of sterile insects to
reduce fertility of a field population of the same species (IPPC, FAO).
• Similarly Sterile Insect is defined as ―An insect as a result of an appropriate treatment is unable to
produce viable offspring (FAO).
• SIT has been known for its eradication of New World Screworm fly, Cochliomyia hominivorax.
• The Idea of this technique was conceived by Dr. E. F Knipling.
It was in the year 1954-55 that Screworm fly got successfully eradicated from Curacao Island. Similar
results were achieved from USA, Mexico and Libya. For this Dr. Edward F. Knipling and Dr. Raymond C.
Bushland were awarded with World Food Prize (1992)

There are four
components of Sterile
Insect Technique
1. Mass Rearing
2. Sterilization
3. Release
4. Monitoring
Components of SIT

Fig: Wild type population and Sterile insect technique release population work in a field condition

Generation Natural
population
of Female
(Assumed)
Sterile Male
insect
Released
S:F Male
ratio
Infertile
Progeny(%)
No. of
female in
each
generation
1 1000 2000 2:1 66.7 333
2 333 2000 6:1 85.7 47
3 47 2000 42:1 97.7 1
4 1 2000 2000 99.9 0
Knipling’s SIT Model
Knipling (1955) also emphasized on following prerequisites before developing and applying SIT which
includes
• Estimates of natural population of target insect must be accurate
• Rear enough sterile insects to over flood natural population.
• The released insect must be distributed uniformly
• Irradiation must produce sterility without affecting competitive mating ability and longevity of insect.
• Female should mate only once.
• If females mate frequently then males should also mate frequently

Various SIT programmes followed
S.no Sterile Insect technique against Insect Pests Countries Reference
1. New World Screwworm, Cochliomyia hominivorax USA Mexico Libya Lindquist et al., 1992
2. Mediterranean fruit fly, Ceratitis capitata Various parts of Latin America Hendrichs et al., 1995
3. Codling moth, Cydia pomonella Canada, USA and Switzerland IAEA, 2001 and Thacker 2002
4. Tsetse fly, Glossina palpalis Zanzibar island, Tanzania
Joint FAO/IAEA Division www.iaea.or.at:80/programs
5. Onion fly, Delia antiqua Netherlands Thacker (2002)
6. Pink bollworm, Pectinophora gossypiella California, USA Thacker (2002)
7. Boll weevil, Anthonomous grandis Lousiana, USA Thacker (2002)
8. House mosquito, Culex quinquefasciatus Florida, USA Thacker (2002)
9. Malarial mosquito, Anopheles ludens El salvador Thacker (2002)

Key social factors influencing uptake of area-wide management (AWM) integrating the Sterile Insect Technique
(SIT), framed as social barriers, facilitators, institutional mechanisms and personal factors.
(Mankad et al., 2017)

• In Integrated pest management, pheromones are used in different ways such as monitoring
through trap catch, killing through mass trapping, mating disruption and attracticide (lure and kill)
methods.
• Pheromone traps in stored insect management can be used to detect both the presence and the density of pests.
• Insects send these chemical signals to help attract mates, warn others of predators, or find food.
• Example: sex pheromones, aggregation pheromones, alarm pheromones, etc.
Pheromones
Fig: Orientation of male moth towards female moth after detecting the pheromone lure

• Push-pull strategies involve the behavioral manipulation of insect pests and their natural enemies via the
integration of stimuli that act to make the protected resource unattractive or unsuitable to the pests (push) while
luring them toward an attractive source (pull) from where the pests are subsequently removed.
• Pushing and pulling is achieved by using repellent (push) and attractant (pull) cues, usually volatiles including
pheromones and allelochemicals.
(Alkema et al., 2019)

Mating disruption with Pheromone traps
• Mating disruption is one concept where synthetic pheromone compounds are employed to
achieve mating failure of insect pests in given area or crop to reduce pest pressure and hence pesticide load
on the crop.

Mechanisms Of Mating Disruption
Disruptants can interfere with mate location in 3 principal ways:
1. Competition: Males may spend time and energy orienting to sources of formulation.
• A variant on this method adds insecticide to point sources of pheromone, an “attract and kill” strategy (Cork 2016).
2. Sensory Impairment:
• Generally, such impairment can be due to adaptation of either sensory receptors or habituation, which is a central
nervous system phenomenon, or both factors.
3. Camouflage:
• The pheromone plume from a calling female becomes imperceptible amongst the background of disruptant.

(Arif et al., 2017)
Examples of some of the lures:

Genome editing by CRISPR-Cas9:
• Clustered Regularly Interspaced Palindromic Repeats/CRISPR-associated protein 9, simply known as CRISPR/Cas9,
is a useful genetic tool for efficient site-directed genome editing.
• The CRISPR/Cas9 system consists of a Cas9 RNA-guided nuclease and CRISPR RNA (crRNA), which guides the
Cas9 enzyme specifically to the target sequence in the genome.
Functions in insects:
• Unravelling Sex Determination Pathways in Insects
• Site-directed Mutagenesis in Pest Insects to Enable Population Control
• Gene Drive Systems for Population Suppression or Replacement
• Knockig out or knocking down of the gene responsible for movement and feeding in insects.

• The sgRNA directs the SpCas9 protein to bind genomic DNA through a 20-nucleotide sequence and further guides it to
introduce a DSB.
• This DSB causes random mutations when repaired by the error-prone NHEJ pathway or precise gene modification when
repaired by the error-free HDR pathway.
(Wang et al., 2019)
• CRISPR, clustered regularly
interspaced short palindromic
repeat;
• Cas, CRISPR-associated;
• DSB, double-strand break;
• HDR, homology-directed
repair;
• NHEJ, non-homologous end-
joining;
• sgRNA, single-guide RNA

• Insects represent the most successful taxon of eukaryotic life, being able to colonize almost all environments.
• Microbial symbiomes associated with insects, impact important physiologies, and influence nutritional and immune
system status, and ultimately, fitness.
• A variety of bacterial phyla are commonly present in insect guts, including Gammaproteobacteria,
Alphaproteobacteria, Betaproteobacteria, Bacteroidetes, Firmicutes, Clostridia, Spirochetes, Verrucomicrobia,
Actinobacteria, and others.
• Among them, the genus Enterobacter has been recognized as a dominant inhabitant of the gut for several important
insect species, indicating an essential functional role for this taxon.
• Enterobacter is a genus of common Gram-negative, facultatively anaerobic, rod-shaped, non-spore-forming
bacteria of the family Enterobacteriaceae.
Functions of Enterobacter:
• Nitrogen fixation
• Degradation of Plant Cell Wall Components
• Degradation and Biosynthesis of Other Nutrients
• Probiotic Effects of Enterobacter
Insect Gut microbiome

(Qadri et al., 2020)

Functions of gut microbiota in tephritid fruit flies
Essential amino
acids, Protein
synthesis, Egg
production
Recycle urea,
plant derivatives
to protein
Increase male
size, copulatory
success
Carbon, nitrogen
metabolism
Suppress the
pathogenic
bacteria
Reduce rearing
duration,
improved survival
Degrades purines,
polysaccharides to
usable nitrogen
Provides vitamins,
lipids and
aminoacids
Overcome the
host plant
resistance
Resistance to
insecticides
Detoxification of
plant toxins
Work as
diazatrophs for
nitrogen fixation
Larval
development,
pupal weight and
sperm storage

• Wolbachia is an obligate intracellular and maternally transmitted α proteobacteria. They reside in reproductive
tissues of invertebrate hosts.
• They are found in 60 per cent of insect species.
• Wolbachia causes reproductive alteration such as
1. Parthenogenetic development
2. Convert genetic males into females
3. Killing males in early developmental stages
4. Cytoplasmic incompatibility
Cytoplasmic Incompatibility (CI) by Wolbachia
Cytoplasmic incompatibility results in mortality of the embryos produced (Bourtzis 2007)
Unidirectional CI
Wolbachia Infected males mated to uninfected females. It results in 100 % eggmortality
Bidirectional CI
When both males and females carrying incompatible Wolbachia strain mates
Wolbachia induced cytoplasmic incompatibility

➢ Cytoplasmic incompatibility induced by Wolbachia.
➢ There are four different mating combinations between infected
and uninfected males and females.
➢ However, infected males (yellow) mated to uninfected females
produce some embryos with early embryonic lethality,
characterized by defects in early mitotic divisions (CI, lower
left) most often observed as defects in chromosome segregation
during late telophase (nuclei, arrowhead).
➢ These defects are rescued when the same infected males are
mated to infected females (Rescue, lower right).
➢ Wolbachia are seen as small punctuate dots, with high
concentrations associated with astral microtubules.
B. Loppin and T.L. Karr, 2005

• It is a control strategy using genetically engineered insects that have (carry) a lethal gene in their genome (an
organism's DNA).
• Lethal genes cause death in an organism, and RIDL genes only kill young insects, usually larvae or pupae.
Male homozygous insect for dominant lethal which was reared under permissive condition when released in the wild population to mate with
wild female then F1 progeny is produced, since these progenies are heterozygous for dominant lethal gene so this gene will express and
cause mortality as also permissive condition is not present under natural condition. Permissive condition like tetracycline in the diet
suppresses the expression of dominant lethal gene in Homozygous male

Figure: Principle of the release of insects carrying a dominant lethal gene (RIDL).
(A) scheme of the transgene.
The tetracycline activator variant (tTAV) protein binds to its own promoter, activates its own transcription and perturbs
overall gene expression in the cells, resulting in mosquito death, unless tetracycline that binds and inactivates tTAV is
provided.
(B) During mass rearing in the production unit, mosquitoes develop normally in the presence of tetracycline. For an
intervention, males are sorted at the pupal stage (based on the smaller size of male pupae).
Once released, they mate with wild females whose progeny will die due to unrestricted tTAV activity

➢ Male annihilation involves the trapping of male fruit flies using a high density of trapping stations
consisting of a male lure combined with an insecticide (usually technical malathion or spinosad), to
reduce the male population to such a low level that mating does not occur.
Male Annihilation Technique (MAT)

Models to be followed in AW-IPM:
Fixed area model
• the control area is fixed in size and there
is no advancing pest control front,
• there is a core area to be protected and a
buffer zone on all sides of the core area.
Rolling carpet model
• the control area is expanding according
to the ―Rolling-carpet principle
• there is a buffer on only one side and
pest free zones on the other sides.
• Both these models consist of two components such as a biological component (i.e., dispersal) and an economic
component (break-even analysis).
• The dispersal part describes the movement of the insects across the buffer zone and will determine the width
of the buffer zone.
• The economic component of the model will, given a certain width of the buffer zone determined by the
dispersal part, allow a calculation of costs and revenues of the control program and will determine the break-
even size of the core area at which control costs equal revenues.
(Barclay et al., 2011)

• The first is the core area, in which the aim is to reduce (in case of a suppression strategy) or eliminate the
pest species.
• The core area contain the actual resource of value, but in other cases, removal of the pest from the core area
may simply have a strategic value by protecting crops situated elsewhere or by protecting humans or
livestock against disease vectors (in case of a containment or a prevention strategy).
• The second is a buffer zone that borders the core area on one or more sides and within which control
methods attempt to kill the target insects within that zone, including those that enter the zone from
outside.
• The buffer zone is defined as the region of an AWPM program that is large enough to prevent the pest insect
from moving from outside the buffer to the core area before being destroyed by the control methods operating
within the buffer zone.
Basic spatial elements of an AW-IPM program:

Dimensions of the area under control:
A is the core area;
T is the total rectangle (core + buffer);
x is the width of the A area;
kx is the length as a multiple of the width;
d is the width of the buffer zone (B) (B = T − A).

Schematic diagram of the expected changes in pest density from an infested area (high pest pressure),
through the buffer zone (B), into the core area (A) in the case of an eradication strategy.
• In the rolling-carpet approach, declines in pest density represent declines over time, although the form of the
slope is schematic.

Case studies

Assessment of the Sterile Insect Technique to Manage Red Palm
Weevil Rhynchophorus ferrugineus in Coconut
R. KRISHNAKUMAR and P. MAHESWARI
Department of Entomology, College of Agriculture, Vellayani - 695522, Thiruvananthapuram,
Kerala, India
Objective: To develop the SIT for use against the red palm weevil on Poothuruth
Island near Dalavapuram Island in Kerala.

Area:
• Poothuruth Island near Dalavapuram Island in Kerala
Sterilization:
• Male red palm weevils were irradiated immediately after their emergence from cocoons, since their sperm
remains immature and vulnerable to dominant lethal mutations when exposed to gamma radiation.
• Irradiation was carried out in a gamma radiation chamber (model 900) with a capacity of one litre and at a
dose rate of 1 Gy/16 seconds, which was ascertained by Fricke dosimetry.
The entire study was conducted in two phases:
(1) through initial laboratory studies to determine the optimal dose of radiation for sterilizing insects, and
(2) trial releases of sterile male weevils in a coconut garden to ascertain the effectiveness of the method in
the field.
R. KRISHNAKUMAR and P. MAHESWARI (2007)

Type of
infestatio
n
Development stage
I
Instar
II Instar III
Instar
IV
Instar
V Instar VI
Instar
VII
Instar
VIII
Instar
IX
Instar
Prepupae
and
pupae
Adults
Crown
infestatio
n
2.22
(1.79)
1
1.09
(1.45)
9.96
(3.31)
1.78
(1.67)
1.72
(1.65)
3.93
(2.22)
14.29
(3.91)
14.29
(3.91)
19.97
(4.58)
29.91
(5.56)
2.96
(1.99)
Stem
infestatio
n
0.72
(1.31)
2.50
(1.88)
2.57
(1.89)
0.56
(1.25)
6.73
(2.78)
9.76
(3.28)
13.83
(3.85)
12.10
(3.62)
19.34
(4.51)
19.97
(4.58)
3.75
(2.18)
Bole
infestatio
n
4.48
(2.34)
4.36
(2.32)
3.75
(2.18)
1.28
(1.51)
8.98
(3.16)
10.56
(3.40)
12.54
(3.68)
26.56
(5.25)
9.96
(3.31)
12.91
(3.73)
6.18
(2.68)
Critical
difference
0.55 0.68 0.55 0.39 0.69 0.65 0.63 0.91 1.05 0.43 0.28
The number of red palm weevil individuals of each life stage present in three types of infested palms (n = 25 for
each type) that were dissected from different red palm weevil infested-coconut plantations of Kerala during 2000-
2001

Number of sterile male red palm weevils (first generation) released based upon the estimated population density
of wild weevils in Poothuruth Island near Dalavapuram, Ashtamudi Lake in the Kollam district of Kerala.

Average number of female red palm weevils captured per trap together with native,
sterilized males, or both during each 20-day period after release

Number of eggs oviposited by native female palm weevils, before and after the release of sterile
insects in each 20-day period after release.
The rate of sterility induced in the native female palm weevil population (as indicated by the percentage egg
hatch) before and after the release of sterile insects in each 20-day period after release.

Estimated number of female red palm weevil present on the island as indicated by mating status
(with native or sterile males) on indicated days after release of sterile males.

Population development as revealed by trap catches of female palm weevils after seven release sessions (first
generation release).
Number of sterile male red palm weevils (second generation) released based upon the estimated population density of wild
weevils in Poothuruth Island near Dalavapuram, Ashtamudi Lake in the Kollam district of Kerala.

Average number of female red palm weevils captured per trap together with native, sterilized
males, or both during each 20-day period after release (second generation release).
Number of eggs oviposited by native female palm weevils, before and after the release
of sterile insects in each 20-day period after release (second generation release).

The rate of sterility induced in the native female palm weevil population (as indicated by the percentage egg hatch)
before and after the release of sterile insects in each 20- day period after release (second generation release).
Estimated number of female red palm weevils present on the island as indicated by mating status (with native
or sterile males) on indicated days after release of sterile males (second generation release).

Population development as revealed by trap catches of female palm weevils after seven release sessions
(second generation release).
Number of sterile male red palm weevils (third generation) released based upon the estimated population density
of wild weevils in Poothuruth Island near Dalavapuram, Ashtamudi Lake in the Kollam district of Kerala.

Conclusion
• SIT was used against the red palm weevil as a component of an AW-IPM strategy.
• When the weevil population is low, the SIT can be an effective method of management of the pest.
• However, with higher weevil populations, suppression methods of pest management such as pheromone traps and
chemical control measures should be carried out to reduce pest population before initiating SIT release.

Community approach for implementation of eco-friendly IPM technology for fruit
fly management in fruits and vegetables in agri-export zones of south Gujarat
Organization:
• Rashtriya Krishi Vikas Yojana is the key to support state
and district action plans funded by the Ministry of
Agriculture, GOI.
• Navsari Agricultural University
Technology Used:
• Male Annihilation Technique (MAT) by using sexual
lures
Management technique:
• NAU has designed and commercialized an eco-friendly,
economical and easily adoptable fruit fly trap popularly
known as "Nauroji-Stonehouse Fruit Fly Trap“ in
2008.
NAU, 2008

Sr.
No.
District No. of
Villages
No. of
Farmers
Area (ha) No. of
Traps
Mango Sapota Cucurbit
vegetables
Navsari
1 Gandevi 32 2325 890 952 - 20612
2 Chikhali 55 3799 1136 719 395 25091
Valsad
3 Valsad 35 2822 1390 304 - 21343
4 Parda 28 1864 1406 223 - 20001
5 Dharampur 37 2280 814 - 162 11481
6 Kaparada 22 2249 819 - 272 12112
Total 209 15339 6485 1196 579 110642
Table: Beneficiaries of villages and number of farmers using the traps against fruit fly in two districts of south Gujarat
Implemented during the year 2008-09 and 2009-10
NAU, 2008

Male and female fruit flies Infected and healthy fruits of Mango
NAU, 2008

Result:
Sr. No. Crop Infestation In per centage Per cent
yield
increased
Treated
Orchards
Untreated
orchards
1 Mango 3.06
(0 to 4%)
30.34
(30 to 35 %)
27.27
2 Cucurbits 2.5 - 4.6
(0 to 4%)
19 - 32
(30.50%)
27
• This Project costs around Rs 7.86 crores and benefitted farmers to the tune of 49 crores.
• An estimated benefit of Rs 81,840 per hectare is achieved by spending a mere Rs 350.
Benefit: cost (233.8: 1).
Conclusion:
NAU, 2008

CRISPR/Cas9 mediated knockout of the abdominal-A homeotic gene in
the global pest, diamondback moth (Plutella xylostella)
(Huang et al., 2016)
Objective: Gene function studies based on genome editing and developing novel approaches
for genetic control of the globally important pest insect diamondback moth (Plutella xylostella)
Yuping Huang,Yazhou Chen, Baosheng Zeng , YajunWang, Anthony A. James, Geoff M. Gurr,
GuangYang, Xijian Lin, Yongping Huang And Minsheng You

Materials and methods:
Experimental DBM strain: The experimental DBM strain (Fuzhou-S) was derived from insecticide-
susceptible insects collected from a cabbage (Brassica oleracea var. capitata) crop in Fuzhou (26.08°N,
119.28°E)
Cloning of Pxabd-A gene

Figure:
(A) Gene structure of the P. xylostella
abdominal-A ortholog (Pxabd-A).
(B) Phylogenetic tree of abd-A based on the
alignment of amino acid sequences of 12
species.
The tree involves three major branches:
Insecta, Myriapoda and Cestoidea
Phylogenetic tree constructed using maximum likelihood method

Figure: qRT-PCR-based expression of Pxabd-A at different developmental stages and sexes of the adult. Statistically-significant
differences were labeled with different letters or letters in parentheses as analyzed with one-way ANOVA (Duncan’s multiple
range test, P < 0.05, n = 3).
Abbreviations:
• E, eggs;
• L1, L2 -1st, 2nd instar larvae;
• L3M, L3F, L4M and L4F - 3rd and
4th male/female instar larvae,
respectively;
• PPM and PPF - male and female
prepupae;
• PM and PF -male and female pupae;
• M and F - male and female adults.

Figure: Phenotypes of Pxabd-A G0 chimeric mutants.
(A) Wild-type 1st instar larvae of P. xylostella showing
three pairs of thoracic appendages located on the
thoracic segments (T1-3, white arrowheads) and four
abdominal appendages on four of the nine abdominal
segments 21 (A1-10, yellow arrowheads);
(B), (C) and (D) shows disorder of body in 1st instar
larvae (red arrowheads), 4th instar larvae and pupae,
respectively.
Wild-type: WT;
CRISPR-treated - disruption of Pxabd-A individuals;

(E) The difference of prolegs between WT and G0 mutants.
The red arrows signify the black crochet disappeared from some prolegs in CRISPR-treated mutants;
(F) Illustration for formed testis in A5-A6 abdominal segments of the 4th instar male larvae.
The red arrows show the position of testis.
The wild-type testis of larvae is bacilliform mainly presenting in A5 and potentially extending to A6.
CRISPR-treated male larvae show defective shapes of testis9

Figure: CRISPR-treated male adults were sterile and abnormal genitals.
(A) The external genitalia of wildtype and G0 mutated male adults.
The red arrows indicate that the external genitalia of all mutated males were deviated from the original location;
(B) The internal genitalia (testis) were highly abnormal.
Testis of the wildtype male shows one regularly spheroidal (left lane).
The red arrows indicate that irregular spherical testis (middle lane) in CRISPR-treated males, and some have two spheroidal
testes (right lane). (Huang et al., 2016)

Conclusion:
• Mutations of Pxabd-A were transmissible to the progeny indicating the feasibility of the
CRISPR/Cas9 system in non-model organisms
• CRISPR/Cas9 mediated genome editing for P. xylostella gene function studies is still challenging
because most genes are recessive so only homozygous mutants display phenotypes

Successful Area Wide Eradication Of The Invading Mediterranean
Fruit Fly In The Dominican Republic
J. L. Zavala-lópez, G. Marte-diaz And F. Martínez pujols
Objective: To eradicate the Mediterranean fruit fly in the Dominican Republic
Technology used:
• SIT-Sterile insect technique to release male insects
• Pheromone traps for the detection of fruit fly larvae and adults
(Zavala-lópez et al., 2019)

Phases and actions of the eradication process followed during the Mediterranean fruit fly eradication campaign
2015-2017 in the Dominican Republic (dotted line is a theoretical representation of population density)

Maximum number of traps used in the national Mediterranean fruit fly
trapping network established in 2015 in the Dominican Republic

Numbers of installed traps (solid bars) and servicing levels of these traps (line) in the eastern
region, including La Altagracia Province, during the 2015-2017 eradication campaign.

Numbers of detected wild adult flies (black bars) and larvae (line) of Ceratitis capitata per week
during the 2015-2017 eradication campaign in the eastern region of the Dominican Republic

Numbers of fruit samples collected (black bars) and Mediterranean
fruit fly larvae detected (brown line) during 2015-2017.

Exports of horticultural products from the Dominican Republic to
the USA between 2011 and 2017, including the export ban in March
2015 because of the Mediterranean fruit fly invasion.

• The last adult Mediterranean fruit fly was detected in the Dominican Republic in the second week of
January 2017.
• Eradication of the pest from the Dominican Republic using an IPM approach including area-wide SIT
application was confirmed in April 2017 after a period of at least three full life cycles with zero
captures.
• The official declaration of eradication took place in July 2017 after six generations of zero catches and
an additional verification trapping network established in high risk areas, including previous detection
sites
Conclusion

Technology Used By Field Managers For Pink Boll Worm Eradication With Its
Successful Outcome In The United States And Mexico
R. T. Staten And M. L. Walters
R. T. Staten And M. L. Walters (2020)
Objective: To eradicate a key pest of cotton pink boll worm over a large geographic area integrating sterile
insect technique in contiguous infested areas i.e., Chihuahua, Sonora, and Baja California in Mexico and also in
the states of Texas, New Mexico, Arizona, and California in the USA.

• The programme covered all activities including extensive GPS mapping, pheromone trap monitoring
for adult populations, and the integration of all control operations.
• Control tools included Bt-cotton, the release of sterile moths, pheromone mating disruption, cultural
control, and on a very limited basis conventional insecticide application.
• Critical area-wide resistance management using sterile moth release, rather than planting susceptible
cotton in refugia, was pioneered in this programme.
Materials and methods:

Figure: Pink bollworm eradication phases, dates, and areas in south-western USA and north-western Mexico

ENTITIES IN USA CONTRIBUTIONS
USDA-APHIS All sterile insect production, USA release
cost, and USA regulatory enforcement
The producer communities: Within-state cost of all non-SIT2 in-field
treatments and operations (includes Bt-
cotton, pheromone mating disruption, and
insecticides)
1. Texas Boll Weevil Foundation
(TBWF)1
All field management of treatments,
monitoring, evaluation and reporting
2. New Mexico PBW and BW
Foundation1
3. Arizona Cotton Research and
Protection Council (ACRPC)
4. California Cotton Pest Control Board
(CCPCB), funds managed by CDFA
Brief outline of management entities involved in the USA and their contributions to the
pink bollworm eradication programme

ENTITIES IN MEXICO CONTRIBUTIONS
SAGARPA (Ministry of Agriculture, Livestock,
Rural Development, Fisheries and Food),
SENASICA (National Service of Health, Food
Safety, and Agriculture Quality)
Leadership, Technical and managerial support,
critical funds (varied year to year dependent on
needs and availability at national level)
USDA-APHIS-International Services and Plant
Protection and Quarantine (PPQ)
Technical and information technology support,
logistical support, bi-national coordination,
coordination with USA embassy for security,
procurement of some supplies, and some field
personnel and SIT2 coordination
1. Comité Estatal de Sanidad Vegetal (state plant
protection committee) de Chihuahua1
State level management of operations (treatment,
survey, and control), funding via grower
assessments and direct contributions
protection committee) de Sonora1
protection committee) de Baja California
R. T. Staten
And M. L.
Walters (2020)
Brief outline of management entities involved in the Mexico and their contributions to the pink bollworm eradication
programme

Technology used:
All activities were sub-divided into three activities:
1. mapping and data management,
2. surveying (trapping and larval sampling), and
3. control.
1. Precise GPS locations of all fields with unique identification numbers for every field and its trap or traps
2. Barcoded identification of all traps with GPS location within the programme
3. Storage and access to all trap and capture data for sterile and non-sterile specimens
4. Precise location of all Bt and non-Bt cotton (Gossypium hirsutum L.) fields, including a distinction for Pima cotton, Gossypium barbadense L.
5. Access to detailed information on all programme-applied pheromone mating disruption treatments, conventional insecticides, and sterile moth
releases – this included access to needed regulatory notifications within each state and flight recordings for all spray and sterile release aircraft,
and
6. Reports generated from complete data by servicing date or any other needed time interval and geographically-defined parameter.
The use of this harmonized system expedited communication within and between state programmes.
Mapping and data management:

Survey Technology
Trap Selection and Use
• Delta trap used
• Throughout the PBW programme, trap density standards were set at one trap per 80 acres (32.4 ha) in the USA and one trap per
20 ha in Mexico for all Bt-cotton (cotton genetically modified to express the endotoxins of Bacillus thuringiensis Berliner) (Bt).
• All cotton fields which did not express these resistant traits were trapped at one trap per 10 acres (4.05 ha) in the USA or one
trap per 4 ha in Mexico.
Trap Lure Formulation
• The discovery and development of the female sex attractant of the PBW was the single most important entomological
breakthrough of the mid 1970’s with respect to PBW control. The name “gossyplure” and its characteristics were first
published by Hummel et al. in 1973.
• Gossyplure is a near 50/50 ratio mixture of (ZZ) and (ZE)-7,11 hexadecadien-1-ol acetate isomers.
Moth Identification
• The programme had to face two critical issues, namely species identification (taxonomic) and separation of sterile from native
insect specimens. From the first sterile moth releases in the San Joaquin Valley in 1968, moth taxonomic identification used
labial palp bands, and genital clasper characteristics to separate P. gossypiella males from other species.
Larval sampling:
• Two different sampling methods were used. Bolls collected from the field could be processed within boll holding boxes (Fye
1976) or by direct examination of bolls cut open immediately after field collection.

Control Technologies
Transgenic Cotton
Mating Disruption
• Within this eradication programme, mating disruption was used on all non-Bt cotton during at least the first four years
of each state’s operations. The hand-applied PBW Rope was preferred.
• Aerially applied NoMate Fiber, NoMate Mec (Scentry Biologicals) and Check Mate (Suterra), were also used when
circumstances required. These latter formulations had an effective disruption time of 8 to 14 days.
Sterile Insect Release
• Releases of sterile moths in this programme had two purposes: a suppression tactic in and of itself, and as a resistance
prevention strategy
• The release of sterile PBW was started in 1968 in the San Joaquin Valley of California as part of a
containment/exclusion strategy to prevent establishment of the pest (Staten et al. 1993, 1999).
• Releases were continuous in areas of detection from 1970 through 2011.
Conventional insecticides
Cultural control

Summary data pink bollworm programme in Texas 2000
through 2004
Summary data pink bollworm eradication programme in
Texas through 2005-2012

Summary data pink bollworm eradication programme for
the Ascensión area of the state of Chihuahua
Summary data pink bollworm eradication programme for
the Meoqui area of Chihuahua

Summary data pink bollworm eradication programme
for the Ojinaga area of Chihuahua
Summary data pink bollworm eradication
programme for the Juárez area of Chihuahua

Summary data pink bollworm eradication programme in New
Mexico (Phase I)
Arizona (Phase II - Arizona Zone 1)

Arizona (Phase IIIa - Arizona Zone 2)
Arizona (Phase IIIb - Arizona Zone 3)

southern California
Northern Sonora, Mexico pink bollworm
eradication programme summary data (Phase IIIb)

Mexicali valley, Baja California, Mexico pink bollworm
eradication programme summary data

Conclusions
• On November 22, 2012 ten municipalities in north-western Chihuahua were declared free of PBW (as officially
eradicated).
• Ascensión work area which had not had a detected population for 5 years. Subsequently, on December 8, 2014,
eradication was declared for the remainder of the state of Chihuahua.
• On February 3, 2016, PBW was declared eradicated from Sonora and Baja California (SENASICA 2018).
• The state of Tamaulipas, which is contiguous to Texas and has likewise been involved in PBW eradication
activities, also had no PBW captures in 2018, but has not yet been declared PBW-free (SADER 2018).
• In the USA, eradication could only be declared after Bt-cotton labelling issues for refugia (grower variety
selection) were resolved.

Area Wide Management Of Mediterranean Fruit Fly With The Sterile Insect Technique In
South Africa: New Production And Management Techniques Pay Dividends
J-H. Venter , C. W. L. Baard and B. N. Barnes
Technology used:
• SIT(Sterile insect technique)
• Sterile males produced with genetic sexing strain VIENNA 8 having more quality based on the temperature
sensitive lethal (tsl) mutation
• Fruit fly densities in commercial orchards are monitored with Chempac® bucket traps baited with a three-
component lure (Biolure) that are deployed at a density of 1 trap per 20 ha
• “Attract and kill” bait stations limited to backyards and hotspots on farms.
Objective: To manage some of the fruit production areas of south Africa as areas of low
pest prevalence by Mediterranean fruit fly.
(Venter et al., 2021)

Figure: Average numbers of wild Mediterranean fruit flies/trap/day (FTD) trapped in three fruit production
areas under SIT application during the first 20 weeks of the year (= harvest period) from 2007/2010 to 2017
• When comparing the average FTD for 2007-2008
(period before the MoU) with that of 2015-2017,
the FTDs in the Hex River Valley decreased by
73% from an average of 4.32 to 1.14. This
average includes hotspots that are focally
supressed.
• The same comparison for the Elgin/Grabouw area
indicates a population reduction of 19% (although
the reduction from the 3-year period immediately
following the MoU is 32%), from a FTD of 0.50
to 0.41.
• When comparing the average FTD for 2010-2011
with that of the period 2015-17, the FTDs in the
Warm Bokkeveld decreased by 78% from an
average of 1.46 to 0.32.
(Venter et al., 2021)

Area Wide Fruit Fly Programmes In Latin America
P. Rendón and W. Enkerlin
Technology used:
• SIT(Sterile insect technique)
P. Rendón and W. Enkerlin (2021)
Factors that Contribute to Pest Movement and Establishment
• Global Trade and Transport
• Human Movement and Travel
• Climate Change
Organizations involved:
• IPPC- International Plant Protection Convention
• FAO- Food and Agriculture Organization
• IAEA- International Atomic Energy Agency

Introductions, establishment and spread of non-native tephritid
fruit fly species in the Americas.

Fruit fly AW-IPM programmes in the LAC region

Putting The Sterile Insect Technique Into The Modern Integrated Pest
Management Tool Box To Control The Codling Moth In Canada
C. Nelson , E. Esch , S. Kimmi , M. Tesche, H. Philip And S. Arthur
(Nelson et al., 2021)
Objective: To integrate chemical, cultural and biological techniques that
complement the SIT into orchard and regional pest management programme
and/or individual growers.

OKANAGAN-KOOTENAY STERILE INSECT RELEASE PROGRAMME (OKSIR programme)
Program area:
• Approximately 600 km2 , and at its onset serviced approximately 8 900 ha (22 000 acres) of pome fruit
• The large area to be serviced, and the need for pre-release sanitation, required the programme to be
implemented sequentially across three zones.
• Pre-release sanitation and construction of the rearing facility began in zone 1 in 1992 followed by moth
release in 1994.
• Pre-release sanitation started in 1998 and 2000 in zones 2 and 3, respectively, with moth release occurring after
two years of sanitation efforts.
Program services:
• The OKSIR programme services include pre-release sanitation, mandatory SIT application, surveillance,
enforcement and education

Figure: Location of the OKSIR programme. The map of Canada (right) indicates where the
OKSIR Programme is located in British Columbia, and the inset (left) illustrates how the
programme area, covering a linear distance of ca. 175 km, was divided into three zones.

Pre-Release Sanitation:
• The first phase of the programme was pre-release sanitation. This entailed the removal of thousands of
unmanaged/abandoned host trees to reduce refugia for the codling moth.
• The programme also coordinated and supported the suppression of codling moth populations in orchards through the use of
conventional insecticides, cultural practices and pheromone-mediated mating disruption.
• Wild codling moth populations had to be reduced as much as possible throughout all communities to increase the efficiency
of subsequent SIT application.
Mandatory SIT Application
• The programme delivers a mandatory area-wide control application of sterile codling moths to every orchard property, i.e.
2000 sterile codling moths of mixed sex (1:1)/ha/week for approximately 20 weeks per season.
Surveillance
• Every orchard property is monitored with pheromone-baited traps (1 trap/ha) that are checked once a week.
• Other monitoring techniques include: in-season fruit inspections, end-of-season assessment of fruit damage, and banding of
host trees (corrugated cardboard strips wrapped around trees to trap mature larvae; later the strips are removed and
destroyed). SIDDU LAKSHMI PRASANNA 94

Results:
• The dashed line indicates the recommended threshold (two codling moths per trap/week for two consecutive weeks) at
which insecticide controls supplementary to the SIT would be required.
Mean wild codling moth captures per trap per week from 1995 to 2017

Percent of programme area with >0.2% of fruit damaged by the codling moth.
The dashed line indicates 10% of the programme area, an economic target set by the Programme’s Board;

Estimated pesticide active ingredient (kg or L) applied per ha per year for all zones
managed by the SIR programme from 1991 to 2016

Conclusions:
• The OKSIR programme clearly illustrates that area-wide integration of the SIT can successfully manage codling
moth populations in an environmentally sound way.
• In addition, it can easily be integrated with other biological control methods such as pheromone-mediated
mating disruption and CpGV.
• Most importantly, the SIT can replace control products that are no longer environmentally or economically
viable, and hence provide an excellent biologically sustainable solution for controlling insect pests.

Biological control:
DEVELOPMENT AND AREA WIDE APPLICATION OF BIOLOGICAL CONTROL USING
THE PARASITOID Aphidus Gifuensis AGAINST Myzus Persicae IN CHINA
• Aphidius gifuensis Ashmead (Hymenoptera: Braconidae) is an important endoparasitoid of many aphids.
• The augmentative use of this parasitoid has achieved area-wide suppression of M. persicae in tobacco and other
crops in China. This approach is being applied on large areas, covering more than 3 million ha between 2010 and
2015.
• This technology has effectively controlled the aphid on tobacco, while other beneficial insects have increased in
the absence of insecticide applications, further protecting biodiversity in the fields and providing long-term
ecological benefits and solved insecticide resistance problems
Y. B. Yu, H. L. Yang, Z. Lin, S. Y. Yang, L. M. Zhang, X. H. Gu, C. M. Li And X. Wang

Comparison of costs of biological and chemical control of aphids

Biological Control: Cornerstone Of The Area Wide Integrated Pest
Management For The Cassava Mealy Bug In Tropical Asia
K. A. G. Wyckhuys , W. Orankanok , J. W. Ketelaar , A. Rauf , G. Goergen And P. Neuenschwander
• The cassava mealybug Phenacoccus manihoti Mat.‐Ferr. (Hemiptera: Pseudococcidae) is a globally important pest
of cassava (Manihot esculenta Crantz), a crop that is cultivated on nearly 25 million ha across the tropics.
• The endophagous parasitoid Anagyrus lopezi De Santis (Hymenoptera: Encyrtidae) introduced in Thailand in
2009
• The host-specific A. lopezi effectively suppresses the cassava mealybug across a range of agro-climatic, biophysical
and socio-economic contexts in tropical Asia, and constitutes a central component of area-wide integrated pest
management (AW-IPM) for this global pest invader.

Benefits of wide area management:
• It enables many producers to pool resources to utilize technologies and expertise that
are too expensive for individual.
• It can avoid external costs due to coordination.
• Cost of pest monitoring, detection and suppression been minimized.
• Helpful in firm communication and decision making of risk management of pest
control
• Automatic contribution of legal authority as it is absolutely essential.

Conclusions:
• The use of chemical insecticides has become increasingly complex due to pest resistance,
environmental concerns, and restrictions on residue levels by importing countries.
• In the interests of reducing insecticide use, as well as pre- and post-harvest crop losses, while
maintaining sustainable agricultural systems, AW-IPM programmes integrating the SIT have
proved effective in supporting safe and environment-friendly international trade.
• Making a community as pest free area through this technology
• This also used in Live stock pest management and human diseases vectors etc.,

AW-IPM.pdf

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