Dr. Christine Schwake-Anduschus, Dr. Norbert HaaseDepartment of Safety and Quality of Cereals

1. Bilder:©CarloSchrodt,VerenaN.,MichaelBührke/PIXELIO
Networking on Aflatoxins
Dr. Christine Schwake-Anduschus, Dr. Norbert Haase
Department of Safety and Quality of Cereals

2.  Filamentous fungi
 Soil microorganism
 Dependent on the inclusion of high-energy organic
nutrients
 Development dependent on aw (water activity) and pH of the
substrate
 Pronounced secondary metabolism
 Co-occurrence of mycotoxins possible
 Example: fumonisines and aflatoxins
2
Fusarium sp.
Aspergillus ochroceus
source: MRI
Mycotoxins
Filamentous fungi
15.08.16Max Rubner-Institut, Department of safety and quality of cereals 2

3. Filamentous fungi and the corresponding mycotoxins
 Fusarium spp.
 Claviceps spp.
 Alternaria alternata (u.a. spp.)
 Aspergillus flavus >25°C
 Penecillium (aurantiogriseum, viridicatum,
verrucosum, hordei)
 Aspergillus (candidus, flavus, fumigatus)
 DON deoxynivalenol often
 ZON zearalenone middel
 T2/HT-2 Toxin seldom in wheat
 fumonisins frequent in maize
 ergot alkaloids middel in rye
 alternariol, AME etc. seldom
 aflatoxins
 ochratoxin A
 aflatoxins
>25°C
3
Frequency in
GermanyIn the field
During storage
Source: BMEL
aw water aktivity << compared to field
15.08.16Max Rubner-Institut, Department of safety and quality of cereals 3

4. structures of mycotoxins
 secondary metabolites
 chemicaly different
 differ in detection
 solubel in water or insolubel
 DON deoxynivalenol
 ZON zearalenone
 T2/HT-2 toxin
 fumonisines
 ergot alkaloid
 alternaria toxins
 ochratoxin A
 aflatoxins
O
OH
CH3
CH2OH
CH3
H H
O
OH
H
O
stabil ~ 120 °C
Bilder: MRI
415.08.16Max Rubner-Institut, Department of safety and quality of cereals 4

5. Time course of aflatoxin formation and reduction in maize
15.08.16Max Rubner-Institut, Department of safety and quality of cereals
Formation: via insect damage,
drought stress
•Slow drying
•Poor storage
Reduction:
•Analysis
•Rejection of lots
•Nixtamalization (cooking with Ca(OH)2 lime , dehulling)
•Milling 5
John I.Pitt et al. Food Control 32 (2013) 205-215

6. 15.08.16 6
Associated partners
 JKI Federal Research Centre for Cultivated Plants
 FLI Federal Research Institute for Animal Health
 PAEPARD Platform for African – European Partnership
 EAFF Eastern Africa Farmers Federation
 KALRO Kenya Agricultural and Livestock Research Organization
 ILRI International Livestock Research Institute
 AflaStop Project funded by the ACDI/VOCA organization
Max Rubner-Institut, Department of safety and quality of cereals
The goal is to initiate a long-lasting scientific network between African and German
partners by working out the possibilities to reduce aflatoxins in the food value chain
via close contacts.
Project leader MRI Federal Research Institute of Nutrition and Food
Dr. Christine Schwake-Anduschus
Budget 158.644 – Duration: 13 month 01.07.16 – 31.07.17
AflaNet Project - Partners

7. Workpackages of AflaNet Project
15.08.16 7Max Rubner-Institut, Department of safety and quality of cereals
WP 2
Applicability
of rapid test
kits for
aflatoxin
detection
WP 3
Aflatoxin M1
development:
Knowledge
about the ratio
B1
feed/M1milk
WP 4
Aflatoxin B1
inactivation of
biosynthesis
at the
molecular
level
WP 6 dissemination of project results,
Networking East-Africa and Kenya-Germany
Prospects of Minimization of Aflatoxin in Feed and Food in Kenya
Long lasting Cooperation of
German Federal Institutes and Kenyan/ East-African Key Actors
Installation of sustainable relationships
WP 1
Liasion officer in Kenya
Excursion and
Experts survey on site
Conference and
exchange
with scientists and
active groups in Kenya,
e.g. EAFF, KALRO, ILRI,
PACA, MRI, JKI, FLI
Germany Kenya
WP 5 Capacity building:
visits of Kenyan scientists at MRI

8. At the beginning of the project a study tour to various stakeholders in the
agricultural production chain, visits of acreages for maize and visits to
research institutions and laboratories are planned.
The liaison officer together with the associated organizations and the
coordinator will plan and organize this 6 day visiting tour starting in
Nairobi.
The aim is to evaluate which groups and projects are working on aflatoxin
minimization and which parameters of minimization strategies for
aflatoxins in maize and dairy products are already established and
applied.
In addition, approx. 10-12 representatives from research institutions,
projects, stakeholders of crop production, storage and processing
facilities as well as from consumer protection associations are invited to
participate in a one-day-conference in order to generate the necessary
further research (~ May/June 2017)
15.08.16 8Max Rubner-Institut, Department of safety and quality of cereals
Workpackages of AflaNet Project

9. Workpackages of AflaNet Project
15.08.16 9Max Rubner-Institut, Department of safety and quality of cereals
WP 2
Applicability
of rapid test
kits for
aflatoxin
detection
WP 3
Aflatoxin M1
development:
Knowledge
about the ratio
B1
feed/M1milk
WP 4
Aflatoxin B1
inactivation of
biosynthesis
at the
molecular
level
WP 6 dissemination of project results,
Networking East-Africa and Kenya-Germany
Prospects of Minimization of Aflatoxin in Feed and Food in Kenya
Long lasting Cooperation of
German Federal Institutes and Kenyan/ East-African Key Actors
Installation of sustainable relationships
WP 1
Liasion officer in Kenya
Excursion and
Experts survey on site
Conference and
exchange
with scientists and
active groups in Kenya,
e.g. EAFF, KALRO, ILRI,
PACA, MRI, JKI, FLI
Germany Kenya
WP 5 Capacity building:
visits of Kenyan scientists at MRI

10. 15.08.16 10Max Rubner-Institut, Department of safety and quality of cereals
Workpackages of AflaNet Project
Analysis
R
ejection
Controlling the goods is one important lever for food security

11. Example of a test system
 Easy to use
 Easy to prepare the samples:
 Extraction (water, ethanol, included in test kit)
 Application of a spezific volume on the strip
 Strip into the reader (or mobile device)
 Automatic determination
 Results/Numbers have to be interpreted easily
 Little information and training required
15.08.16Max Rubner-Institut, Department of safety and quality of cereals 11
Handheld device
Test strips

12. What MRI/GE can do
Max Rubner-Institut, Department of safety and quality of cereals
- More elucidations
- Develop and hand out recommendations
- Training and science transfer
- Suitability testing of test systems
3 test kit systems have to be tested
Which one?
Requirements for Kenya?
1215.08.16

13. Workpackages of AflaNet Project
15.08.16 13Max Rubner-Institut, Department of safety and quality of cereals
WP 2
Applicability
of rapid test
kits for
aflatoxin
detection
WP 3
Aflatoxin M1
development:
Knowledge
about the ratio
B1
feed/M1milk
WP 4
Aflatoxin B1
inactivation of
biosynthesis
at the
molecular
level
WP 6 dissemination of project results,
Networking East-Africa and Kenya-Germany
Prospects of Minimization of Aflatoxin in Feed and Food in Kenya
Long lasting Cooperation of
German Federal Institutes and Kenyan/ East-African Key Actors
Installation of sustainable relationships
WP 1
Liasion officer in Kenya
Excursion and
Experts survey on site
Conference and
exchange
with scientists and
active groups in Kenya,
e.g. EAFF, KALRO, ILRI,
PACA, MRI, JKI, FLI
Germany Kenya
WP 5 Capacity building:
visits of Kenyan scientists at MRI

14. 15.08.16 14Max Rubner-Institut, Department of safety and quality of cereals
What MRI/GE can do
Experiments with oral administration of aflatoxin B1 by bolus (accurate
dosing) and the administration of naturally contaminated feed (field
conditions) are planned. The daily intake should be at 10 or 50
micrograms per cow per day and not exceed the limit of 5 micrograms
/ kg aflatoxin B1 for single feed. Analysis of milk is carried out
Produce milk product below the official limits in the urban centers and
on the other hand enable the dairy industry to produce milk products
which can be exported to other countries.
Analyze the aflatoxin M1 in the products (cheese, yogurt,..
Binding of aflatoxin have to be checked by adding different probiotic
cultures and looking on the bioavailability of the aflatoxin afterwards.
An evaluation of necessary experiments should be in focused during
this work package after consultations in Kenya

15. Workpackages of AflaNet Project
15.08.16 15Max Rubner-Institut, Department of safety and quality of cereals
WP 2
Applicability
of rapid test
kits for
aflatoxin
detection
WP 3
Aflatoxin M1
development:
Knowledge
about the ratio
B1
feed/M1milk
WP 4
Aflatoxin B1
inactivation of
biosynthesis
at the
molecular
level
WP 6 dissemination of project results,
Networking East-Africa and Kenya-Germany
Prospects of Minimization of Aflatoxin in Feed and Food in Kenya
Long lasting Cooperation of
German Federal Institutes and Kenyan/ East-African Key Actors
Installation of sustainable relationships
WP 1
Liasion officer in Kenya
Excursion and
Experts survey on site
Conference and
exchange
with scientists and
active groups in Kenya,
e.g. EAFF, KALRO, ILRI,
PACA, MRI, JKI, FLI
Germany Kenya
WP 5 Capacity building:
visits of Kenyan scientists at MRI

16. 15.08.16 16Max Rubner-Institut, Department of safety and quality of cereals
What MRI/GE can do
WP 4
Aflatoxin B1
inactivation of
biosynthesis at the
molecular level
ddPCR System for aflatoxin producing fungi will be established as a
very sensitive tool to monitor the presence of A. flavus and/or A.
parasiticus and to analyze the transcription of the aflatoxin
biosynthesis genes under various conditions.
After establishment the system can be used to correlate biomass
and/or gene expression with the aflatoxin actually analyzed in a
sample. These data will be useful for the development of new
measures based on molecular principles to control aflatoxin
biosynthesis.
Together with the expertise of African partners concerning
endogenous maize production treatments and drying technologies,
the influence of various environmental parameters, resulting from
these treatments on the activation of aflatoxin biosynthetic genes
will be analyzed by ddPCR (or qPCR). Especial emphasis will be
laid on the combination between drying and temperature

17. Workpackages of AflaNet Project
15.08.16 17Max Rubner-Institut, Department of safety and quality of cereals
WP 2
Applicability
of rapid test
kits for
aflatoxin
detection
WP 3
Aflatoxin M1
development:
Knowledge
about the ratio
B1
feed/M1milk
WP 4
Aflatoxin B1
inactivation of
biosynthesis
at the
molecular
level
WP 6 dissemination of project results,
Networking East-Africa and Kenya-Germany
Prospects of Minimization of Aflatoxin in Feed and Food in Kenya
Long lasting Cooperation of
German Federal Institutes and Kenyan/ East-African Key Actors
Installation of sustainable relationships
WP 1
Liasion officer in Kenya
Excursion and
Experts survey on site
Conference and
exchange
with scientists and
active groups in Kenya,
e.g. EAFF, KALRO, ILRI,
PACA, MRI, JKI, FLI
Germany Kenya
WP 5 Capacity building:
visits of Kenyan scientists at MRI

18. 15.08.16 18Max Rubner-Institut, Department of safety and quality of cereals
WP 5 Capacity building:
visits of Kenyan scientists at MRI
Workpackages of AflaNet Project
Two African scientists will join the working groups
from WP 2, 3 or 4 for approximately three weeks in
Germany. The persons will be trained in MRI
laboratories to transfer the gathered knowledge and
the so far conducted results to the associated
African partners within the duration of the work
packages. The time of the visit, the work plan and
the question which department of MRI and
institution in Kenya will be involved need to be
discussed at the beginning of the project

19. Workpackages of AflaNet Project
15.08.16 19Max Rubner-Institut, Department of safety and quality of cereals
WP 2
Applicability
of rapid test
kits for
aflatoxin
detection
WP 3
Aflatoxin M1
development:
Knowledge
about the ratio
B1
feed/M1milk
WP 4
Aflatoxin B1
inactivation of
biosynthesis
at the
molecular
level
WP 6 dissemination of project results,
Networking East-Africa and Kenya-Germany
Prospects of Minimization of Aflatoxin in Feed and Food in Kenya
Long lasting Cooperation of
German Federal Institutes and Kenyan/ East-African Key Actors
Installation of sustainable relationships
WP 1
Liasion officer in Kenya
Excursion and
Experts survey on site
Conference and
exchange
with scientists and
active groups in Kenya,
e.g. EAFF, KALRO, ILRI,
PACA, MRI, JKI, FLI
Germany Kenya
WP 5 Capacity building:
visits of Kenyan scientists at MRI

20. 15.08.16 20Max Rubner-Institut, Department of safety and quality of cereals
Thank you for your attention
Questions?