This project has developed new ‘Smart’ Spore and Insect Trapping systems for target(s) surveillance referenced to GPS and climate data (temp, wind direction, RH), or wireless data transmission for improved compatibility to rapid and accurate downstream diagnostics.

1. SESSION 2: MORE ACCURATE AND TIMELY DIAGNOSTICS
TO BETTER INFORM BIOSECURITY DECISIONS
Rohan Kimber

2. 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)

3. 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

4. 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

5. Next generation trapping (2015 - )
 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

6. Systems developed/being evaluated by PBCRC project 2014
Collaborative effort
Innovation - Prof Jon West (Rothamsted Research)
Engineering
– Stuart Wili, Burkard Manufacturing Co (UK) – Jet Sampler
– Les Zeller, Paul Kamel, Bruen Smith, USQ – NCEA – SMART’s
technology
Smart Spore Traps - meteorological triggers eg. Temp, RH, diurnal, wind
speed/direction
- Impactor type (capture on adhesive type) – CRC30032
- Miniature Virtual Impactor (direct capture to tube)
Jet Spore Sampler
- Virtual impaction system, high sampling rate, 20-tube carrousel system
- Mobile trapping - Smart capture (GPS)
Burkar
d
Burkar
d
USQ
Burkard
New technology & platforms – Airborne spore surveillance – PBCRC Project 2014

7. New technology & opportunities – Jet Spore Sampler
Jet Spore Sampler – Burkard Manufacturing Co.
 Prototype 1 – delivered to SARDI – Mobile Jet Spore Sampler
 SMART technology adaptation being finalised at USQ
 GPS and climate data (temp, wind direction, RH)
 Prototype 2 – delivered to Rothamsted Research (UK) Prof Jon West – Jet Spore Sampler
 Fixed (static) location air sampling.
 For evaluation in Rothamsted wind tunnel – optimise operational function and
collection efficiency
 That version will not be a mobile device.
 Visits supported by PBCRC Small Initiative Projects SI20004 & SI20060

8. New technology & opportunities – Jet Spore Sampler
Jet Spore Sampler – Evaluation at Rothamsted Research (July 2016) with Prof Jon West
 Prototype 2 – determine collection efficiency in comparison to Burkard Volumetric Trap
 Lycopodium clavatum (used in aerobiology studies) approx 35um diam
 Giant puffball spores (Lycoperdon giganteum) approx. 3.5um diam
Standard vs Multi-vial vs Jet Spore
Samplers Rothamsted Wind Tunnel
July 2016
Lycopodium
clavatum
Lycoperdon
giganteum

9. 1970s Jet Spore Sampler
(samples onto culture plates
or plant leaves)
2016 Jet Spore
Sampler fan
assembly 2016 Jet Spore Sampler
(samples into diagnostic
tubes)
New technology & opportunities – Jet Spore Sampler
Jet Spore Sampler – Modifications at Burkard Manufacturing Co (Stuart Wili) July 2016
 Prototype 2 –
 Initial tests showed poor collection efficiency (<20%) compared to standard (impactor) trap
 New settings & components fitted – replicating 1970’s sampler – optimising spore deposition into tube
 Duplicated components now fitted to Australian prototype

10. New technology & opportunities – Jet Spore Sampler
Jet Spore Sampler – Evaluation at Rothamsted Research (July 2016) with Prof Jon West
Wind tunnel assessments – comparison to standard volumetric spore trap
 35 um size spores in wind speed 3 m/s the JSS was on average 70%
 35 um size spores in wind speed 6 m/s the JSS was on average 98%
 3.5 um size spores in wind speed 3 m/s the JSS was on average <1%
 3.5 um size spores in wind speed 6 m/s the JSS was on average <1%
Conclusions
 The Jet Sampler is effective in moderate to high wind speed sampling (3-6 m/s).
 Well suited as a Mobile Jet Spore Sampler, sampling air from a moving vehicle at approx. 100
km/h.
 Captures particles approx. 35 um diam. very efficiently. Requires mass to separate from air
column
 Many airborne fungal spores identified for targeted surveillance for the grains industry
between 20-100um (including cereal rust spores).

11. New technology & opportunities – Mobile Jet Spore Sampler
Smart Mobile Jet Sampler – Spores
 Smart capture – GPS, temp, RH, time
 12V power - vehicle
 Dry or liquid capture – 2ml DNA tubes
 High sampling rate – 450 l/min
 16-tube carrousel - automated
Burkard
Intake
Carouse
l driver
Suction = 450
l/min
Vacuum
motor16-tube
carouse
l
Sampl
e tube
Temp/R
H
sensor
Controller
12V
supply &
Bluetooth
Roof-
rack
Separatio
n
chamber
Exhaust

12. Burkard
New generation Mobile Jet Sampler
• Vehicle mounted eg. courier/school bus
• Assign sampling grids within route
(‘Smarts’)
• Index to corresponding vial within each GPS
grid
• Samples (vials) sent to lab for analysis
1
2
3
45
6
7
8
GPS grid sampling example
Jet Sampler –
1980s
Mobile Jet Sampler –
2016
New technology & opportunities – Mobile Jet Spore Sampler

13. Mobile/Network surveillance of air-borne spores
Where to next ..
• Field testing against standard impactor trap – molecular vs microscopic
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

14. 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

15. Thank you
Burkard
 Jon West (Rothamsted Research)
 Stuart Wili (Burkard Manufacturing Co.)
 Helen DeGraaf (SARDI – Entomology)
 Greg Baker (SARDI – Entomology)
 Dr 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)
 Bruen Smith - USQ