aflasafe development for aflatoxin mitigation in tanzania national steering commitee meeting

www.iita.org A member of CGIAR consortium

aflasafe

Development for Aflatoxin

Mitigation in Tanzania

Ranajit Bandyopadhyay: IITA

Peter Cotty: USDA-ARS

On behalf of the Aflasafe Team

National Steering Committee for Mycotoxins in Tanzania,

Dar-Es-Salaam, 1 Aug 2014

• 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

acute

acute hepatic necrosis, cirrhosis,

carcinoma

Death; 108 in 1974 in Gujarat, 250

to 15,000 ppb aflatoxins in corn

chronic

carcinogenic

associated with stunting

anti-nutritional

immune-suppressive

gut integrity?

BBC 2004, Gong et al 2004, NIEHS 2010

underreported

unknown

Human Health Effects

www.iita.org

Aflatoxin Contamination Occurs in

Two Phases

Phase I: Before Crop Maturity

Developing crops become infected.

Associated with crop damage (insect, bird, stress).

Favored by high temperature (night) and dry conditions.

Phase II: After Crop Maturity

Aflatoxin increases in mature crop.

Seed is vulnerable until consumed.

Rain on the mature crop increases contamination.

Associated with high humidity in the field & store, insect

damage, and improper crop storage or transportation.


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

Kenya (CDC and Kenyan Ministry of Health 2004)

District Samples

% samples with aflatoxin levels (ppb)

<20 21-99 100-1,000 >1,000

Makueni 91 35 13 40 12

Kitui 73 38 21 32 10

Machakos 102 49 25 23 3

Thika 76 66 17 13 4

Total 342 47% 19% 27% 7%

Tanzania (IITA & partners, unpublished, 2013)

Aflatoxins in Markets

Crops Samples % samples with aflatoxin levels (ppb)

<10 11-100 101-1,000 >1,000

Groundnut 180 89 11 0 0

Maize 287 71 15 10 4

Cassava 405 86 10 4 0.2

Aflatoxin and Poultry (Broilers)

Aflatoxin

levels in

feeds in

Nigeria

Aflatoxin level (ppb) Samples (%)

<20 (safe) 38

>20 to 100 (up to 5x) 14

>100 to 500 (up to 25x) 41

>500 to 1,000 (up to 100x) 7

AF-free diet 500 ppb AF diet

AF-free

diet

500 ppb AF diet

~40% reduction in live weight (8 weeks)


EPA approved 2 products

AF36

Afla-guard

More than 1 million hectares 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!


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)

Thus, aflatoxin contamination reduced

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 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


Aflatoxin & Aspergillus

A. flavus

L-strain

Aflatoxin: +/-

A. tamarii

Aflatoxin: -

A. flavus

S-strain

SBG

Aflatoxin: +++

A. parasiticus

Aflatoxin: ++++


Atoxigenic Strain Identification

Collection/characterization Toxin assay

Field

efficacy

test

Lab

competition

assay

VCG/DNA characterization

cnx nia-D

Unknown 2

+


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

www.iita.org

• IITA

• USDA

• AATF

• BMGF

• Doreo Partners

• National institutions

Strong Partnership


Broadcast @ 10 kg/ha 2-3 weeks before flowering

Sporulation on moist soil

Spores

Insects

Aflasafe in 5 kg boxes

3-20 days

Wind

Soil colonization

30-33 grains m-2

Fungal network in killed grain

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)


0

10

20

30

40

50

60

2010 2011

Aflasafe™

Control

0

20

40

60

80

100

120

140

160

180

2010 2011

Aflasafe™

Control

82 94 82 93

16 82 16 81 Fields (#)

At Harvest After storage


Aflato

xin

(ppb)

Aflatoxin Reduction

(%)

*

Nigeria: Efficacy on Groundnut


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


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


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)


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


Aflasafe Development in Tanzania

• 510 samples (260 maize grain and 250 groundnut kernel) samples from Tanzania

• 5017 isolates obtained from these samples. Happy Daudi (ARI-Naliendele) trained

• 818 atoxigenic isolates identified

• 690 isolates characterized by SSR and 136 by CAPs

• 20 atoxigenic isolates belonging to 12 haplotypes; 6 also occur in Kenya

• Testers developed for these 20 atoxigenic isolates

• Being complemented with 444 toxigenic and 284 atoxigenic nit- mutants


Tanzania Haplotypes

Haplotype

Number of Samples

Tanzania

(239 samples)

Kenya

(357 samples)

Mozambique

(38 samples)

Zambia

(97 samples)

BOWAGA 40 4 0 0 BAQEGA 31 0 1 0 BOHEZI 16 0 0 0 BOPAJI 13 1 0 1

BOHAMA 11 1 0 0 BAQIFO 14 0 0 0 BOHEGI 7 8 0 1 BAQEZE 8 2 0 1 BOHAJU 6 0 0 1 BOHIPO 4 0 0 1 BAQALE 6 4 0 1 BAQESO 13 0 0 0


Proposed Plan

• 8-12 isolates will be selected for first field trial in Tanzania

• To be conducted in 120 farmers’ fields in Mtwara, Dodoma and Manyara in 2014/15 season

• Field trials will be managed in collaboration with national partners (suggestions required) and private sector (e.g., Export Trading Group, BRITZ, and the Clinton Development Initiative) .

• Samples will be analyzed in IITA-Dar and IITA-Ibadan

• 4 most efficient isolates selected to constitute Aflasafe TZ01

• Large-scale evaluation of Aflasafe TZ01 in 2015 and 2016

• Registration, business plan, manufacturing from 2016 onwards


Product Types National

Products

Products ready for registration

Products under testing

Strain development in progress

Aflasafe-NigeriaTM

Aflasafe-SenegalTM

Aflasafe-KenyaTM etc…

Aflasafe-WestTM

Aflasafe-EastTM

Aflasafe-SouthTM

Regional

Products Senegal

Mali

Burkina

Ghana

Nigeria

Kenya

Tanzania

Mozambique

Zambia


Challenges

• Aflatoxin is a hidden problem

• Chemical analysis required

• Awareness is low

• Long incubation for expression of health impacts

• 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……

www.iita.org www.iita.org A member of CGIAR consortium

Integrated Management

The elements are:

• Technology

• Awareness – entire range of value chain participants

• Advocacy – regional, national, investors

• Training – farmers, transporters, traders, regulators, consumers

• Policies – standards, harmonization, trade

• Institutions – regulators, markets, testing, private sector

• Trade / Markets – food/feed processors, poultry/fish industry

• Public good – home consumption; urban and rural markets; government procurement, HGSF


Farmer Training


Markets

Doreo Partner analysis

Poultry industry

Export-oriented aggregators

Food processors

Large commercial farmers

Smallholder farmers

Market based

• Poultry feed

• Premium food

market

• Export

• AgResults (Incentive-

cum-market based)

• Public distribution

ma

rke

t d

em

an

d f

or

Afl

as

afe

• 60% maize consumed by farmers

• 40% sold in the market

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

Pilot Implementation


• Value chain-centric: Farmers’ and other value chain participants’ interest as the foundation of the action

• Public sector intervention with health perspectives for smallholders

• Action-oriented: Using practical methods to actively solve problems, not just talking about ideas, plans, or theories

• Innovation platform: problem solving by participants working regularly together to address common issues and challenges.

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


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%

• Mean recovery of aflasafe strains from samples (n = 88): 72% to 89%

• Samples with >70% aflasafe strains

(n = 88): 65% to 100%

• Aflasafe maize kept for family (n = 60): 46%


Aflasafe Return on Investment

Quantity sold

(tons) Premium

(%) Premium

(USD)

Aflasafe cost

(USD)

Finance cost

(USD)

Net profit (USD)

Seasonal RoI

AgResults Premium

(USD)

Total Profit (USD)

Total RoI

120.0 7.5% 3,000 836 146 2,017 241% 1,800 3,817 456%

150.0 7.5% 3,750 1,046 183 2,521 241% 2,250 4,771 456%

96.0 3.6% 1,200 669 117 414 62% 1,440 1,854 277%

128.0 13.2% 5,600 892 156 4,552 510% 1,920 6,472 725%

32.0 7.5% 800 223 39 538 241% 480 1,018 456%

30.1 1.8% 188 210 37 -58 -28% 452 393 187%


Scaling-Out

• Nigeria: AgResults • Senegal: Area-wide treatment in

2013 • Kenya: Government buy-in;

excellent support • Zambia: Large-scale efficacy tests

and demonstration of product value with private sector

• Need for business plan, manufacturing capacity, marketing and distribution strategies

• Critical role of PACA and RECs

Coming in 2014-2015…

The Gambia, Benin,

Togo, Ivory Coast,

Uganda, Rwanda,

Burundi, Ethiopia, South

Sudan, Malawi


Aflasafe Production in Lab 60 tons produced in 2011, 2012, and 2013 for deployment


Aflasafe Manufacturing Facility

Large-scale: capacity 5 tons/hour

La

Capacity Development

Laboratory infrastructure

• Burkina Faso (Austria)

• Kenya (USDA/USAID)

• Zambia (USAID)

• Mozambique (USDA/USAID)

Human Technical capacity

• Senegal

• Ghana

• Cameroon

• Tanzania

• Nigeria

Training of African biocontrol researchers, USDA-ARS, Univ. of Arizona

(funded by USDA-FAS with a grant from USAID)

A Zambia technician

receiving training on

aflasafe production at

IITA, Ibadan (USAID)

• 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, AATF & 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/

aflasafe development for aflatoxin mitigation in tanzania national steering commitee meeting

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