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Networking on Aflatoxins
Dr. Christine Schwake-Anduschus, Dr. Norbert HaaseDepartment of Safety and Quality of Cereals
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
15.08.16 Max Rubner-Institut, Department of safety and quality of cereals 2
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
15.08.16 Max Rubner-Institut, Department of safety and quality of cereals 3
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
OOH
CH3
CH2OH
CH3H H
O
OH
H
O
stabil ~ 120 °CBilder: MRI
415.08.16 Max Rubner-Institut, Department of safety and quality of cereals 4
Time course of aflatoxin formation and reduction in maize
15.08.16 Max 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
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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 FoodDr. Christine Schwake-Anduschus
Budget 158.644 – Duration: 13 month 01.07.16 – 31.07.17
AflaNet Project - Partners
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 3Aflatoxin M1 development:Knowledge about the ratio B1 feed/M1milk
WP 4Aflatoxin B1 inactivation ofbiosynthesis 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 KenyaLong lasting Cooperation of
German Federal Institutes and Kenyan/ East-African Key Actors
Installation of sustainable relationships
WP 1Liasion officer in KenyaExcursion andExperts survey on siteConference and exchangewith 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
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
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 3Aflatoxin M1 development:Knowledge about the ratio B1 feed/M1milk
WP 4Aflatoxin B1 inactivation ofbiosynthesis 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 KenyaLong lasting Cooperation of
German Federal Institutes and Kenyan/ East-African Key Actors
Installation of sustainable relationships
WP 1Liasion officer in KenyaExcursion andExperts survey on siteConference and exchangewith 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
15.08.16 10Max Rubner-Institut, Department of safety and quality of cereals
Workpackages of AflaNet Project
Analys
isReje
ction
Controlling the goods is one important lever for food security
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
15.08.16 Max Rubner-Institut, Department of safety and quality of cereals 11
Handheld device
Test strips
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 testedWhich one?Requirements for Kenya?
1215.08.16
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 3Aflatoxin M1 development:Knowledge about the ratio B1 feed/M1milk
WP 4Aflatoxin B1 inactivation ofbiosynthesis 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 KenyaLong lasting Cooperation of
German Federal Institutes and Kenyan/ East-African Key Actors
Installation of sustainable relationships
WP 1Liasion officer in KenyaExcursion andExperts survey on siteConference and exchangewith 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
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
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 3Aflatoxin M1 development:Knowledge about the ratio B1 feed/M1milk
WP 4Aflatoxin B1 inactivation ofbiosynthesis 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 KenyaLong lasting Cooperation of
German Federal Institutes and Kenyan/ East-African Key Actors
Installation of sustainable relationships
WP 1Liasion officer in KenyaExcursion andExperts survey on siteConference and exchangewith 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
15.08.16 16Max Rubner-Institut, Department of safety and quality of cereals
What MRI/GE can do
WP 4Aflatoxin B1 inactivation ofbiosynthesis 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
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 3Aflatoxin M1 development:Knowledge about the ratio B1 feed/M1milk
WP 4Aflatoxin B1 inactivation ofbiosynthesis 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 KenyaLong lasting Cooperation of
German Federal Institutes and Kenyan/ East-African Key Actors
Installation of sustainable relationships
WP 1Liasion officer in KenyaExcursion andExperts survey on siteConference and exchangewith 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
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
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 3Aflatoxin M1 development:Knowledge about the ratio B1 feed/M1milk
WP 4Aflatoxin B1 inactivation ofbiosynthesis 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 KenyaLong lasting Cooperation of
German Federal Institutes and Kenyan/ East-African Key Actors
Installation of sustainable relationships
WP 1Liasion officer in KenyaExcursion andExperts survey on siteConference and exchangewith 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
15.08.16 20Max Rubner-Institut, Department of safety and quality of cereals
Thank you for your attention
Questions?