Scaling up aflatoxin biocontrol in Africa

Agriculture for Nutrition & Health www.iita.orgA member of CGIAR consortium
Scaling-Up
Aflatoxin
Biocontrol in
Africa
Ranajit Bandyopadhyay
IITA, Ibadan, Nigeria
Peter Cotty
USDA-ARS, University of Arizona
Aflasafe Team
1st African Symposium on Mycotoxicology
Livingstone, Zambia, 26-28 May 2015

• Highly toxic metabolite
produced by the ubiquitous
Aspergillus flavus fungus
• The fungus resides in soil and
crop debris, infects crops and
produces the toxin in the field
and in stores
• Death, liver cancer, immune-
suppression, stunted growth
• Impacts animal productivity
• Negatively impacts trade
• Fungus carried
from field to store
• Contamination
possible without
visible signs of the
fungus
Aflatoxin Facts
Photo: Peter Cotty

Aflatoxin
Intervention
Medical
Agriculture
Surveillance
Enterosorption
Pre-harvest
Food processing
e.g. Novasil clay
Agriculture & Medical Prevention of Aflatoxin-
related Food Security and Health Effects
(Adapted from Wild and Hall, Mutation Res., 2000)
Awareness
Regulation
Provision of safe food
Early diagnosis
Post-harvest

www.iita.orgA member of CGIAR consortium
Pre-Harvest Problem
Aflatoxin
(ppb)ppb)
Peanut (n = 188) Maize (n = 241)
Distribution (% samples)
> 4 54 70
> 10 41 52
> 20 29 24
Descriptive statistics (ppb)
Minimum < LOD < LOD
Maximum 3487 838
Mean 111 33
LOD = Limit of Detection; 1 ppb
Aflatoxin in Groundnut and Maize at Harvest
Increases in store
Agriculture for Nutrition & Health

EPA approved 2 products
AF36
Afla-guard
Hundreds
of
Thousands
of acres
treated
annually
in the US!
Production Room
Atoxigenic Strain Manufacturing Facility
Arizona Cotton Research & Protection Council
(Funded and Governed by the Farmers of Arizona),
Phoenix, Arizona
It Works in
Africa Too
Biocontrol Works!

www.iita.org
• IITA
• USDA
• AATF
• BMGF/USAID
• Doreo Partners
• National institutions
Strong Partnership
www.iita.orgA member of CGIAR consortium Agriculture for Nutrition & Health

Biocontrol Principles
 In nature, some strains produce a lot
(toxigenic), and others no aflatoxin
(atoxigenic) (Donner, Soil Biol Biochem
2009)
 Atoxigenic strains are already present on
the crop (Atehnkeng et al., IJFM, 2008)
 Increase the frequency of atoxigenic
strains to competitively displace
toxigenic strains (Cotty & Bayman,
Phytopath 1993) to reduce aflatoxin
contamination
 Atoxigenic strains can be applied without
increasing infection and without
increasing the overall quantity of A. flavus
on the crop or in the environment (Cotty,
Phytopath 1994; Atehnkeng et al., Biological
Control 2014)
0
1
2
3
4
5
6
7
8
9
0 20 40 60 80 100
AflatoxinB1(ng/gX10,000)
Isolates (%) in Applied Atoxigenic Strain
 Strains move from
field to stores
 Multiple year & crop
carry-over effect
(Jaime & Cotty,
Phytopath 2006)
 We use only native
strains

Strain Selection Criteria
In the laboratory (~5,000 strains):
• Does not produce aflatoxin
• VCG/SSR group with
Wide geographic distribution
No toxigenic member
• Defective in >2 aflatoxin & CPA
genes
• Outcompetes toxigenic strains
After field application:
• Superior capacity to colonize,
multiply and survive in soil
• Superior frequency of isolation
from grains
• Superior capacity to reduce
aflatoxin
8-12 native strains
selected for field tests
4 native strains
formulated into
the final
product

Broadcast
@ 10 kg/ha 2-3 weeks
before flowering
Sporulation on moist soil
Spores
Insects
Aflasafe in 2.5 & 5 kg bags
3-20
days
Wind
Soil
colonization
30-33 grains m-2
How Does aflasafe Work?

Efficacy Trials: Data Collection
• All trials conducted in farmers’
fields on crops grown by farmers
• Aflasafe applied by farmers
• Soil sampled before treatment
and grains at harvest:
– Aspergillus population density
– Aspergillus strain profile
– Incidence of aflasafe strains
• Aflatoxin concentration in grains
at harvest and after poor storage

Design & Analysis of Trials
• Field trial size: 0.25 to 15 ha
• Number of fields: 14 to 200
per year
• Paired plot: Each treated
field with its own
companion control field in
close vicinity
• Each farmers’ field
considered as a replicate
• Student’s t-test to compare
treatment effects

Nigeria: Efficacy on Maize
372
0
20
40
60
80
100
120
2009 2010 2011 2012
Aflasafe™
Control
0
100
200
300
400
500
600
2009 2010 2011 2012
82 94 83 86 82 93 89 90
51 14 199 38 51 14 166 38Fields (#)
Less (%)
At Harvest After Storage
*All means of aflasafe and control pairs significantly different; Student’s t-test (P<0.05)
*
Aflatoxin(ppb)

Area Sample Treatment
Mean
Aflatox
(ppb)
Reduct.
(%)
Mean
Aflatox
(ppb)
Reduct.
(%)
Mean
Aflatox
(ppb)
Reduct.
(%)
Diourbel
Harvest
Treated 1.9
93
6.6
87
3.7
82
Control 29.7 50.1 20.3
Storage
Treated 4.4
86
2.1
91
6.9
81
Control 31.3 22.1 35.5
Nioro
Harvest
Treated 4.4
75
5.6
76
5.4
90
Control 17.6 23.1 55.7
Storage
Treated 3.5
95
2.8
94
11.5
84
Control 52.1 46.7 72.5
*All means of aflasafe treated and control pairs significantly different; Student’s t-test (P<0.05)
Senegal: Efficacy of aflasafe SN01
2010 (n=40) 2011 (n=34) 2012 (n=71)

Basis of efficacy: species shift
Treatment
(n = 14)
Aspergillus species/strain distribution (%) – MAIZE/NIGERIA
Soil before inoculation Grain at harvest
L SBG parasiticus L SBG parasiticus
Aflasafe™ 90 aB 7 aA 3 aA 100 aB 0 bA 0 aA
Control 78 aB 15 aA 7 aA 83 bB 16 aA 0.3 aA
Means within the column with different lowercase letters are significantly different according to the t-
test at 5% level of probability. Means within the row with different uppercase letters are significantly
different according to the Fisher’s LSD test at 5% level of probability
Region Treatment
Aspergillus Colony Forming Units/g – G-nut/Senegal
2010 (n = 20) 2011 (n = 17)
Soil Kernel Soil Kernel
Diourbel
Control 2311 a 2912 a 474 a 3257 a
Aflasafe SN01 1793 a 3598 a 795 a 3965 a
Nioro
Control 228 a 3367 a 369 a 3572 a
Aflasafe SN01 120 a 3189 a 470 a 4275 a
*All means of aflasafe and control pairs significantly different; Student’s t-test (P<0.05)
Aspergillus population does not increase due to aflasafe application
No change in Aspergillus Pop.

Bars with same letter within the same
crop/year not significantly different (P<0.05)
Basis of Efficacy: Strain Shift
0
10
20
30
40
50
60
70
80
90
Soil Grain Soil Grain
2009 (n = 49) 2010 (n = 14)
Control Treated
0
10
20
30
40
50
60
70
80
90
Soil Grain Soil Grain
2009 (n = 2) 2010 (n = 16)
Proportion of 4 aflasafe™ strains in soil before treatment
and grains after harvest in control and treated fields
Aflasafestrains(%)
aa a aa a aa a aa a
b
b b
b
Carry-over of inoculum: 71, 52
and 28% after 1, 2, and 3 years

Kenya: Efficacy of aflasafe KE01™
Area (fields) Control Treated
Reduction
(%)
Hola (n = 20) 885 20 98
Bura (n = 16) 105 7 93
Makueni (n = 15) 85 1 99
Aflatoxin (ppb)
*All means of aflasafe treated and control pairs significantly different; Student’s t-test (P<0.05)
38
20
0
88
60
33
0
10
20
30
40
50
60
70
80
90
100
Treated
Control
Fields (%) above
10 ppb in 3 areas
Fields(%)
Deadly (3,700 ppb & 2,270 ppb)
533 ppb
Hola

Product Development in Africa
Senegal
Burkina
Faso
Ghana
Nigeria
Kenya
Tanzania
Mozambique
Zambia
Rwanda
Malawi
Burundi
Uganda
The
Gambia
Strain
development in
progress
Products under
testing in
farmers’ fields
Product ready
for registration
Product
registered

Challenges
• Aflatoxin is a hidden problem
• Chemical analysis required
• Awareness is low
• Long incubation for expression of
liver cancer
• Regulations either non-existent or
poorly enforced
• Market does not usually discriminate
• Demonstration of product value
• Lack of biopesticide manufacturers
The value of a
technology on the
shelf is as much
as the cost of the
space it occupies
on the shelf.
Must translate
knowledge into
usable products
and practices to
benefit people
But……

Key Elements
• Value chain-centric: Farmers’ interest
as the foundation of the action.
Target: Value chain and home-grown
food
• Action-oriented: Using practical
methods which involve doing things
to deal with problems, not just talking
about ideas, plans, or theories
• Innovation platform: An approach to
problem solving through which actors
with a stake in a common issue or set
of issues get together regularly to
address their common challenges.

Poultry Feeding Study
$3,200 net
profit from
10,000 birds
in 8 weeks
www.iita.orgMycored Europe, 28 May, 2013A member of CGIAR consortium
Aflasafe maize feed
Toxic maize feed

Innovation Platform
• Platform meetings with
leadership and members of
Poultry Association of
Nigeria, feed manufacturers,
maize aggregators, aflasafe
farmers, vet professionals
and regulators
• Poultry farmers to buy all
aflasafe maize at a negotiated
premium
• Agriculture ministry to fund
NAFDAC to set up aflatoxin
testing facilities in each state
www.iita.orgA member of CGIAR consortium Agriculture for Nutrition & Health

www.iita.orgwww.iita.orgA member of CGIAR consortium
Integrated approach to manage aflatoxins
Aggregation
Aflasafe
Inputs & training to
improve productivity
Farmer groups/
value chain/Finance
Training for
pre/postharvest
afla management
Awareness and
sensitizations
Policy and advocacy
Market linkages
Aflatoxin testing

G-20 AgResults Aflasafe
• Pull mechanism – Aflasafe is one of the first three pilots
• Provides incentives after demonstrating adoption
• Private sector driven, but focused on smallholder groups
• Implementers provide credit, inputs and technical services to
increase yield
• Aflasafe purchased at cost to improve quality
• Maize tested for aflasafe strains; if present in large frequency,
the implementers incentivized with $18.75/ton maize
• Implementers negotiate maize sale at premium
• Project provides aflatoxin awareness, training of
implementers, and identifies potential market linkages
• Target: 260,000 ha in 4 years

AgResults Aflasafe Pilot -- 2013
Some key statistics
• Number of implementers: 4
• Number of farmers: 1,015
• Treated area: 1,457 ha
• Average productivity: 4.3 tons/ha
• Maize aggregated for sale: 2,031 tons
• Samples with <4 ppb AF (n = 660): 99%
• Samples with >70% aflasafe strains
(n = 88): 65% to 100%
• Premium in market: 1.8% to 13.5%
• ROI before incentive: -28 to 510%
• ROI after incentive: 190% to 725%
• Aflasafe maize kept for family (n = 60): 46%

Scaling-Out
• Nigeria: AgResults farmers to
produce 260,000 tons of
Aflasafe maize; Public-private
partnership
• Senegal: Area-wide treatment
in 2013 and 2014 with 16 tons;
20 tons use projected in 2015;
private sector led
• Kenya: Government buy-in;
about 230 tons procured;
excellent support
• Critical role of PACA and RECs
Senegal
Kenya

Aflasafe Manufacturing Facility
Large-scale: capacity 5 tons/hour
Cost: ~US$1.6 million; Price: $14 - $18/ha
Purpose: Scaling up

Modular Manufacturing Facility
Capacity: 5 tons/day
Cost: ~US$400,000
Purpose: Introduction
Cost: $12 – 15/ha
Labour intensive

Current and future biocontrol efforts
• Create a sustainable system
(commercialization/public good) where
small holder farmers have access to
Aflasafe and are incentivized to utilize
Aflasafe to control aflatoxin levels
• Need for business plan, manufacturing
capacity, marketing and distribution
strategies
• Advocacy, awareness, demonstration of
product value
• Full registration, licensing and
stewardship
• Training and technical back-stopping
• Develop second generation product
• Develop regional strains

• Aflatoxins in food and feed pervasive
in Africa
• Biological control, as the foundation,
with other practices can dramatically
reduce aflatoxin contamination and
improve food safety and security
• Efforts underway to pilot
commercialization of aflatoxin
biocontrol and develop regional
strains
• The pilots need to be up-scaled and
efforts to improve efficacy needs a
fillip for wide-spread impact on
health and trade in Africa
Summary

IITA
Tucson
USDA/ARS
IITA, USDA, & Doreo have Teamed up to Bring
Aflatoxin Prevention to Africa
Made Possible by Many National Partners in Ministries, Industry, and on the Farm
Nigeria
For more information about aflatoxin biocontrol for Africa, check out: www.aflasafe.com

THANK YOU
IITA Business Incubation Platform

Scaling up aflatoxin biocontrol in Africa

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