scaling up aflatoxin biocontrol in africa

Agriculture for Nutrition & Health www.iita.org A 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.org A 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.org A 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 100Afl

ato

xin

B1 (

ng

/g X

10,0

00)

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 38 Fields (#)

Less (%)

At Harvest After Storage

*All means of aflasafe and control pairs significantly different; Student’s t-test (P<0.05)

*

Aflato

xin

(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

Afl

asafe

str

ain

s (

%)

a a a a a a a a a a a a

b b b

b

Carry-over of inoculum: 71, 52

and 28% after 1, 2, and 3 years


http://www.google.co.uk/url?sa=i&rct=j&q=&esrc=s&frm=1&source=images&cd=&cad=rja&docid=lKk0dkUswMQkNM&tbnid=iNOHk0UxXmQVFM:&ved=0CAUQjRw&url=http://naijaintellect.wordpress.com/2012/07/26/nigerian-govt-releases-two-new-hybrid-maize-varieties/&ei=2BeSUsG8EKeH0AXah4DICw&bvm=bv.56988011,d.ZG4&psig=AFQjCNEwOZ2zmi8Rt6wdDo6BOODAz2xl1g&ust=1385392435580697


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

100Treated

Control

Fields (%) above 10 ppb in 3 areas

Fie

lds (

%)

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.org Mycored Europe, 28 May, 2013 A 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.org A member of CGIAR consortium Agriculture for Nutrition & Health

www.iita.org


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

http://www.entwicklung.at/en/


THANK YOU

IITA Business Incubation Platform


scaling up aflatoxin biocontrol in africa

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