The document details a workshop on hyperspectral imaging held in Tulln, Austria, focusing on the detection of contaminants such as ergot in agricultural products using NIR (near-infrared) spectroscopy. It discusses the advantages of hyperspectral imaging over traditional microscopy, including faster analysis times and the ability to detect multiple contaminants simultaneously. Various applications of NIR imaging in agriculture, such as assessing plant resistance and detecting cyst nematodes, are also highlighted.

1. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 20151
Detection of undesirable substances
by NIR hyperspectral imaging spectroscopy
and chemometrics in agriculture
Philippe Vermeulen,
Juan Antonio Fernández Pierna, Pierre Dardenne, Vincent Baeten
(CRA-W Belgium)

2. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 20152
NIR hyperspectral imaging
applications:
Detection and quantification
of contaminants
in the cereals production control sector

3. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 20153
Study case: ergot
• Reemerging of the ergot presence in cereals
– For the farmer, yield reduction (10%)
– For the feed/food sector, high toxicity risk
• Ergot is a sclerotium formed by
the fungi Claviceps purpurea
including ergot alkaloids
• EU legislation:
– 500 mg/kg of sclerotia in food
– 1000mg/kg of sclerotia in feed

4. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 20154
Reference method
• The existing microscopy method provides an
elegant early warning tool for ergot
contamination but is time-consuming
Sorting by visual observation Confirmation by microscope

5. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 20155
Detection based on one mean
spectrum of the sample
Detection based on the analysis
of several subsamples
What can we do using NIRS?

6. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 20156
- Measurement in reflection mode
- Wheel of 30 vials
- Analysis time: +/- 4 hours !!!
for 3 wheels replicated 4 times
by sample
What can we do using NIRS?

7. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 20157
Pure ergot
Bottom
ergot
Positive
vials
Negative
vials
What can we do using NIRS?
PCA on 324 spectra of one sample adulterated at 0,05%

8. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 20158
What can we do using NIRS?
Set of 7 laboratory samples with 0 0,01 0,05 0,1 0,15 0,5 and 1%
of ergot in cleaned wheat

9. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 20159
Contaminants in food
and feed: Inexpensive
detection for control of
exposure
(FP7 project n°211326)

10. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201510
• NIR camera setup
NIR line scan camera: features
- Wavelength range: 1000-2500 nm by step of 6 nm
- 1 line = 320 pixels = 320 spectra
- field of view: 10 cm
- Analysed surface = continuous
- Time of acquisition = 50 millisec/pixel line
- Speed of the conveyor belt = 3 mm/sec
- 1 pixel = 275 µm * 275 µm = 0.075 mm2
1000 1500 2000 2500
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
Wavelength (nm)
Absorbance(Log1/R)
Ergot
Wheat

11. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201511
Ergot detection in wheat by NIR imaging
Wheat
kernels
Ergot
Bodies
The number of pixels counted
for each class of the model
is also provided.

12. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201512
Results of ergot bodies detection
Set of 7 laboratory samples with 0 0,01 0,05 0,1 0,15 0,5 and 1%
of ergot in cleaned wheat
Corrective factor weight/area
Reference value: % weight
Predicted value: % area * 0,8

13. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201513
Peer reviewed Journal article

14. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201514
NIR line scan camera: instrument
in demonstration at NUTRECO
Tray
NIR camera
SWIR
ImSpector
N25E
Spectra
Camera
(Specim Ltd)
Light source

15. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201515
Ergot detection in wheat by NIR imaging
Prediction results of the SIMCA (Soft Independent
Method of Class Analogy) model
The number of pixels counted for
each class of the model and …
(object quantification)
… the distribution of groups
of pixels detected as ergot
are also provided
(object identification)
Wheat
kernels
Ergot
Bodies

16. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201516
Results of ergot bodies detection
Set of 7 samples (2009-2010)
wheat, rye
Set of 6 samples (2011)
rye, triticale, oat
BurgerMetrics
Instrument
(Pilot
imaging system)
SisuChema
Instrument
(Commercial
imaging system)
Corrective factor weight/area
Reference value: % weight
Predicted value: % area * 0,8

17. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201517
ABC special issue: CONffIDENCE outputs

18. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201518
Other applications
• ergot detection in black oat

19. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201519
Other applications
• Multicontaminants detection: ergot, datura, …
Mixture of wheat, black oat, rape seed, ergot and datura

20. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201520
Video

21. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201521
Peer reviewed Journal article

22. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201522
Benefits of the method
for a feed Company
Classical microscopy NIR hyperspectral imaging
High skilled personal Low skilled personal
15 min / 250g A few minute / 250 g
Reduced samples Large samples (sampling more
representative)
Dedicated to ergot Multiple contaminants

23. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201523
NIR hyperspectral imaging
applications:
assessment of resistant plants
in sugar beet breeding program

24. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201524
Nematodes cysts counting
by
NIR hyperspectral imaging system

25. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201525
What is the issue ?
Yield reduction in relation to the cyst
nematode number in the sugar beet
roots
Brown and white cyst
nematode (Heterodera
schachtii)

26. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201526
How to control the contamination?
Cyst nematode development cycle
Tolerant sugar beet plants breeding
Cysts at different development
stages (optical microscope)

27. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201527
Status of the analytical aspects?
Cysts counting by optical microscopy using a
camera system at SES VANDERHAVE
Cysts = red spots

28. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201528
NIR line scan camera: instrument
SWIR ImSpector N25E spectral camera (Specim Ltd)
Conveyor belt (Burgermetrics)
Parameters
- Wavelength range: 1000-2500 nm by step of 6 nm
- 1 line = 320 pixels = 320 spectra
- 1 box = 100 000 spectra
- Time of acquisition for 1 box= 1 min
NIR image acquisition of one box

29. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201529
Spectral libraries
Spectral data preprocessed
1200 1400 1600 1800 2000 2200 2400
-0.1
-0.05
0
0.05
0.1
0.15
0.2
0.25
Variables
Mean
1 Cyst
2 Root
3 Substrat

30. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201530
Data treatment: Applying of 3 discrimination equations and DBSCAN
Image at 1720 nmPicture DQ110053-08 Removing background (Eq1)
Removing substrat (Eq2) Removing root (Eq3) DBSCAN
Discrimination models

31. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201531
DQ110053-08 – Plot 302 - Susceptible plant
Ref value: 80 Predicted value: 80
1 cyst = 4 pixels
Nematodes cysts counting by line scan HIS

32. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201532
DQ110053-20 – Plot309 – Tolerant plant
Ref value: 49 Predicted value: 31
1 cyst = 4 pixels
Nematodes cysts counting by line scan HIS

33. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201533
10 Tolerant plants 10 Susceptible plants
Reference values 40 (24-51) 69 (51-82)
Predicted values 21 (5-49) 58 (26-82)
(1 cyst = 4 pixels)
Nematodes cysts counting by line scan HIS

34. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201534
Cercospora leaf spots assessment
by
NIR hyperspectral imaging system

35. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201535
Samples (DQ100742-01 to 08) Instrument : Line scan HIS
Cercospora leaf spots assessment by line scan HIS
4 cerco tolerant plants
4 cerco susceptible plants

36. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201536
Cercospora leaf spots assessment by line scan HIS
DQ100742-02 : Prediction on-line (PLSDA model)

37. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201537
Cercospora leaf spots assessment by line scan HIS
DQ100742-02 3 days after

38. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201538
Cercospora leaf spots assessment by line scan HIS

39. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201539
Crop residuesToxic plants (Senecio)Animal/vegetal feed
Melamine in soybean meal)
Collagen inside the bones
Collagen (COL+)
No collagen (COL-)
1200 1400 1600 1800 2000 2200 2400
1000
2000
3000
4000
5000
6000
7000
8000
Wavelengths (nm)
Absorbance
Collagen classes: mean spectra
COL+
COL-
Dry fruits
Poster
And many other applications …

40. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201540
Thank you for your attention
Ph. Vermeulen and all the team
Contact:
p.vermeulen@cra.wallonie.be
Acknowledgements

41. SLOPE and BiRT Hyperspectral Imaging Workshop - BOKU-UFT , Tulln Austria, 20th March 201541
http://chimio2016.sciencesconf.org
Contact: chimio2016@cra.wallonie.be
2 Days training: 17-18 January 2016
2 Days conference: 19-20 January 2016