This document discusses a project to develop new tools and technologies for field grain surveillance and diagnostics of high priority exotic pests. It involves developing smart insect and spore traps using sensors, GPS, and wireless transmission. The traps will detect and identify endemic and exotic pests and pathogens. Researchers are testing pheromone stacking, suction traps, and mobile jet spore samplers. Samples will undergo qPCR diagnostics to identify mixed populations. The goal is a coordinated surveillance network using smart sampling and lab-based identification to rapidly detect and report biosecurity threats.

1. biosecurity built on science
Plant Biosecurity Cooperative Research Centre
Project 2014
Part A: New tools for field grains surveillance
Jenny Davidson: Senior Plant Pathologist (SARDI)
Rohan Kimber: Plant Pathologist (SARDI)
Greg Baker: Entomologist (SARDI)
Helen De Graaf: Entomologist (SARDI)
Les Zeller: Engineer (USQ)
Paul Kamel: Engineer (USQ)

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New opportunities for pest & disease surveillance using new technologies & sensing systems
 Broad scale surveillance (coordinated network)
Detect & report endemic or exotic incursions of fungal spores / insects in the field to the decision
maker or grower community
 Local (on farm) outbreaks
 Regional threats (e.g. rusts)
 National interests (biosecurity)
Project 2014 – New tools for field grains surveillance and diagnostics of high priority exotic pests
New technology & opportunities

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New technology & opportunitiesTechnologies implemented
• Species specific detection – qPCR, Pheromone lures (stacking)
• Sensors - optical, impact & digital to confirm target
• Smart capture – GPS, environment or diurnal triggers
• Wireless / telemetry networked grid & transmission
Systems being developed / evaluated –
A. Smart insect traps (moths) – in field detection and imagery
B. Suction insect traps (aphids) – localized monitoring
― Lab qPCR diagnostics (mixed populations)
C. Smart traps (pathogens) – mobile & fixed systems
— Lab qPCR diagnostics & toward ‘real-time’ sensor detection
Field evaluation, technology adaptation, collaboration
• DAFF/USQ & SARDI – Qld and SA test sites
• Rothamsted & Burkard Manufacturing Co. – UK
Research & opportunities – Surveillance and New technologiesNew technology & platforms – pest & disease surveillance – PBCRC Project 2014
B
C
A

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Research & opportunities – Surveillance and New technologies
Smart Insect Trap – Pheromone
 Les Zeller & Paul Kamel (USQ)
 Pheromone – targeted
 Impact & optical sensors (false triggers, oversensitive)
 Digital imagery on automated carousel
 Data transmission (4G telemetry) to a web portal
 Smart data – time stamped
 Three prototypes evaluated
these image (25-80 KB) were
transferred from trap via
telemetry < 1 second
trap
wind
lure attraction range
(pheromone plume)
exotic pest
endemic pest
Non-target pest
Smart Insect trap – Field testing 2016 & 2017
 Helicoverpa punctigera (native budworm)
× Diamond back moth (DBM), requires new algorithm
− Light brown apple moth (LBAM), no flights
o Noctuid (cutworm), due to commence
New technology & platforms – Insect surveillance

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PHEROMONE STACKING
Assess potential of “piggy-backing” existing pheromone trap networks.
Need to test pheromones for compatibility.
Field trial at Waterloo, SA, Spring 2015:
• Turnip moth (As) (Exotic)
• Diamond back moth (DBM) (Established)
• Native budworm (Hp) (Native)
Helen DeGraaf – SARDI

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Research & opportunities – Surveillance and New technologiesNew technology & platforms – Insect surveillance
Mixed Insect Trap – Active Suction (aphid)
 Burkard Manufacturing Co. (UK)
 High sampling rate (>200 l/min)
 240V with 4-8 pot carousel
 1.5m – localised monitoring (aphids)
 Field testing 2015 & 2016 – SARDI
 Proof-of-concept = GPA monitoring (PCR assay Dr Kelly Hill, SARDI)
 Russian Wheat Aphid (exotic target)
Burkard
Mixed Insect Trap – Smart Active Suction (aphid)
 Les Zeller (USQ) prototype 2017
 12V solar powered fan suction (optimised)
 Smart capture – temp, RH, temporal, wind speed or direction
 Mixed populations with carousel system (10 pots)
 Field testing 2017 - SARDI

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Research & opportunities – Surveillance and New technologiesNew technology & platforms – Insect surveillance
• Total Aphid capture at 3 sites in SA 2016
0
20
40
60
80
100
120
140
160
180
3/17/2016 4/17/2016 5/17/2016 6/17/2016 7/17/2016 8/17/2016 9/17/2016
Total numbers of aphids captured at 3 SA sites 2016
Waite (17th Mar) Balaklava (27 May) Kapunda (14 April)
• GPA capture at 3 sites in SA 2016
0
2
4
6
8
10
12
14
16
18
20
Numbers of GPA captured at 3 SA sites 2016
Waite (17th Mar) Balaklava (27 May) Kapunda (14 April)
Helen DeGraaf – SARDI

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Systems being developed/ evaluated – qPCR diagnostics
C
Burkard Manufacturing Co.
Molecular detection - qPCR diagnostics (mixed populations)
• Kelly Hill – GPA* primer design, WFT* published primer
• Helen DeGraaf – GPA & WFT identification & counting
• Molecular Diagnostic Centre – Sample & assay processing
*GPA = Green Peach Aphid WFT = Western Flower Thrip
Target GPA WFT Removed
Mock
mixture
Field sample
0 nil nil nil nil nil
1 -
5 -
10 -
New technology & platforms – Insect surveillance

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• Russian Wheat Aphid (RWA) capture at 3 sites - originally the exotic species target
Helen DeGraaf – SARDI
Research & opportunities – Surveillance and New technologiesNew technology & platforms – Insect surveillance
0
1
2
3
4
5
6
5/4/2016 6/4/2016 7/4/2016 8/4/2016 9/4/2016 10/4/2016
Number of RWA captured at 2 SA sites2016
Balaklava (27 May) Kapunda (14 April)
Diuraphis noxia, alate
Start trap End trap Days trapping Sum of 4 YPTs Kapunda ST Balaklava ST
24/08/2016 1/09/2016 8 0 0 0
1/09/2016 15/09/2016 14 3 2 11
15/09/2016 5/10/2016 20 2 0 16
5/10/2016 19/10/2016 14 2 5 3
Total RWA trapped by yellow pan traps at 4 sites in Tarlee and suction traps at Kapunda and Balaklava (suction trap data [24-48hr] pooled to match YPT trap periods)
Maarten van Helden – SARDI
Yellow Pan Trap – YPTsSuction Trap – ST

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Next generation trapping (2017 - )
 High sampling efficiency & automation systems
 ‘Smart capture’ – GPS, meteorological triggers, data analytics
 More compatible or adaptable to rapid down-stream diagnostics
 Lab-based qPCR – species specific primers for accurate ID
 Toward ‘real-time’ detection (in-field) eg. LAMP, biosensors, fiber optic sensing
 Fixed location
 Low sampling rate = 10 l/min
 Adhesive tape (impact sampling)
 Retrospective analysis – DNA probes
 Several endemic pathogens to pulse & cereals
 SARDI & collaborators – SA, Vic, NSW, WA
Current status – Trapping
Research & opportunities – Airborne spore surveillance

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New technology & opportunities – Jet Spore Sampler
Standard vs Multi-vial vs Jet Spore Samplers
Rothamsted Wind Tunnel July 2016
Lycopodium clavatum Lycoperdon giganteum
Burkard
Systems developed and evaluated by PBCRC project 2014
Collaborative effort
Innovation & testing - Prof Jon West (Rothamsted Research)
Engineering - Stuart Wili, Burkard Manufacturing Co (UK)
Jet Sampler – 1980s
Jet Sampler – 2016

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New technology & opportunities – Mobile Jet Spore Sampler
Smart Mobile Jet Sampler – Spores
 Les Zeller, Paul Kamel at USQ – NCEA
 Smart capture – GPS, temp, RH, time
 Operational refinement – Mobile Trapping
Intake
Carousel
driver
Suction = 450 l/min
Vacuum
motor16-tube
carousel
Sample
tube
Temp/RH
sensor
Controller
12V supply &
Bluetooth
Roof-rack
mounted
Separation
chamber
Exhaust

13. biosecurity built on scienceBurkard
New generation Mobile Jet Sampler
• Vehicle mounted eg. industry/courier/school bus
• Assign sampling polygons (16) within route (‘Smarts’)
• Index to corresponding vial within each GPS sector
• Operational control – Android Tablet (Jet App)
• Samples (vials) submitted to lab for analysis
New technology & opportunities – Mobile Jet Spore Sampler
Mobile Jet Sampler – 2017
GPS grid polygons defined using
Google Earth

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Mobile/Network surveillance of air-borne spores
Where to next ..
• Field testing against standard impactor trap – efficiency & strategies
How should it be used ..
• Within a coordinated (fixed & mobile) trapping network – spatial resolution
i. New smart samplers – identify optimal sampling using model targets
(diurnal or environmental)
ii. Current standard spore traps – adhesive (microscopy)
iii. Future in-field detection systems (lab-in-a-box) – UK collaborators
• Strategic response sampling
i. Deployed sentinel sampler to incursion or to determine point of origin
How could data be managed ..
• Coordinated database management & automated reporting to end-users
i. Molecular Diagnostics Centre (SARDI) – cross sectoral, endemic &
exotic target reporting
Mobile/Network Surveillance of Airborne Spores

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Air samplers
DNA-specific probes
Highly specific, multiplex
qPCR
Smart air samplers – mixed populations
Insect samplesMulti-vial carousel
Traffic light – Biosecurity alerts
Delivery - Pest / Spore Detection & Reporting
http://www.syngenta-crop.co.uk/brassica-alert/
Image analysis / recognition
Data triage & downstream ID
Spores
Pest/Pathogen species composition
Courtesy Jon West - Annemarie Justesen, Aarhus University, Dk
Endemic pathogen dispersal patterns - Spores

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Thank you
Burkard
 Jon West (Rothamsted Research)
 Stuart Wili (Burkard Manufacturing Co.)
 Helen DeGraaf (SARDI – Entomology)
 Greg Baker (SARDI – Entomology)
 Kelly Hill (SARDI)
 Les Zeller (USQ - NCEA)
 Paul Kamel (USQ - NCEA)
 Molecular Diagnostics Centre (SARDI)
 John Weiss (Vic DEDJTR)
 Jamus Stonar (SARDI)
 Tracey Brookes (MEA)
 Sarah Noack (Hart Field Site)
 Matt Tscharke – USQ
 Maarten van Helden - SARDI