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GESTIONE INTEGRATA GESTIONE INTEGRATA DELLE MICOTOSSINE
IN PRE‐ E POST‐RACCOLTO
Antonio F. Logrieco
Istituto di Scienze delle Produzioni AlimentariConsiglio Nazionale delle Ricerche,
ISPA‐CNR, Bari, Italy
V Congresso Nazionale: Le Micotossine nella filiera agro‐alimentare 28‐29‐30 settembre 2015
V Congresso Nazionale: Le Micotossine nella filiera agro‐alimentare 28‐29‐30 settembre 2015
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Similar factors influence the development of disease and resulting
i l i ( l i mycotoxin accumulation (overlapping triangles)
HOST•Susceptibility
h
ENVIRONMENT•Temperature
•Growth Stage•Flowering synchrony
p•Moisture •Radiation
Diseaseand Mycotoxins
PATHOGENImportant differences in the relationships between host PATHOGEN
•Species/population•Density/abundance •Aggressiveness
relationships between host plants, pathogen and
environment may affect disease and toxin differently •Aggressivenessdisease and toxin differently (offsetting the triangles)
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SOURCES OF MYCOTOXIN EXPOSURE
Field fungiSt f i
gStorage fungi . . .
DISTRIBUTIONSTORAGE PROCESSING CONSUMPTIONGROWING CROPS
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MYCOTOXIN MANAGEMENT
Field fungiSt f i
gStorage fungi . . .
DISTRIBUTIONSTORAGE PROCESSING CONSUMPTIONGROWING CROPS
PRE‐ POST‐HARVEST HARVEST
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Ide tif h d• Identify hazards• Find critical pointsp• Establish limitsE t bli h it i• Establish monitoring
• Design corrective actionsg• Verify efficiency
d• Keep records
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MYCOTOXIN MANAGEMENT ASPECTSMYCOTOXIN MANAGEMENT ASPECTS
PRE‐HARVEST POST‐HARVEST
l lComplex(too many factors)
Simple
Good AgriculturalLimited control
Good AgriculturalPractice (GAP) limittoxin accumulation
Problems and side effects of corrective
toxin accumulation
No side effectsactions
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PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
GOOD AGRICULTURAL PRACTICE (GAP)( )
Resistant varieties
Crop rotation
Soil treatmentSoil treatment
In‐planta detoxification
Fungicide application
Biological controlBiological control
Weed and pest management
Agronomic measures
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PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
GOOD AGRICULTURAL PRACTICE (GAP)( )
Resistant varieties
Crop rotation
Soil treatment RESISTANT SUSCEPTIBLESoil treatment
In‐planta detoxification
RESISTANT SUSCEPTIBLE
Fungicide application
Biological controlBiological control
Weed and pest management
Agronomic measures
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Resistant varieties
In resistant maizemost of the genes
Infection withFusarium verticilloides
most of the genes provide a basal
level of defence to the fungus prior to the fungus prior to
infection
Induction of about 80
early genes
In susceptible maizemost of the PR genes
Induction of about 240 late
most of the PR genes are induced after
infection
9
4genes
Lanubile et al.
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Resistant varieties
250
150
200
250
50
100
DON ppm
0
Sum
ey-3
3/81
.60/
/Kő
Wuh
an 6
Bbe
oka
Boz
ut /
MM
/NB
MM
/Sum
3Fr
onta
naM
M/S
um3
t//M
M/N
BP7
318
Ari
naFu
rore
Lud
wig
Ari
stos
Öth
alom
K
imon
Ritm
oW
90Z
11.1
Car
dos
P863
5C
ontr
aZ
ugol
yPe
ntiu
mJ
89.8
1249
Bisc
aySJ
981
1.53
FDK %
Sum
3
NobRSt
Sgv/
NB
//M
Sgv/
NB
//MR
StLSJS
r = 0.7548, P = 0.001
DON content in different wheat cultivarscompared to FHB resistant varieties and linescompared to FHB resistant varieties and lines
Mesterházy, 2014(Leslie and Logrieco Eds.)
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Resistant varieties
/ear
)
120
140
160
180
Thrip
s (n
umbe
r/
40
60
80
100
120
0
20
kg) 120
140Mar Apr M
MAIZE HYBRIDS SELECTION AFFECTS
Fum
onis
in (m
g/
40
60
80
100May SELECTION AFFECTS
FUMONISIN RISK
0
20
(%) 40
Fusa
rium
ear
rot (
10
20
30
Hybrids
A B C
F
0
10
Munkvold, 2014 (Leslie and Logrieco Eds.)
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Indirect effect on mycotoxin levels
Aflatoxins more prevalent in Crops weakened by insect or other
Tolerance to drought stress
pcrops stressed by drought
p ydamage are more susceptible to
mold growth and aflatoxincontamination
Protection against corn borer
contamination
Optimum® AQUAmax® HybridsOptimum® AQUAmax® Hybrids
DuPont Pioneer Hi‐Bred
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PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
GOOD AGRICULTURAL PRACTICE (GAP)
LIMITED SOURCES OF RESISTANCE
NO GM SOLUTION APPROVED IN EU
( )
Resistant varietiesNO GM SOLUTION APPROVED IN EU
RESISTANT VARIETIES NOT ALWAYS HIGH‐YIELDINGCrop rotation
Soil treatmentSoil treatment
In‐planta detoxification
Fungicide application
Biological controlBiological control
Weed and pest management
Agronomic measures
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PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
GOOD AGRICULTURAL PRACTICE (GAP)( )
Resistant varieties
Crop rotation
Soil treatmentSoil treatment
In‐planta detoxification
Fungicide application
Biological controlBiological control
Weed and pest management
Agronomic measures
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Crop rotationEffect of different previous crop on DON
level in wheat plants ?level in wheat plants ?
Clark et al., 2009
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PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
GOOD AGRICULTURAL PRACTICE (GAP)
ECONOMIC CONTRAINTS
( )
Resistant varietiesECONOMIC CONTRAINTS
POLITICAL INCENTIVES
Crop rotation
Soil treatmentSOIL EROSION
Soil treatment
In‐planta detoxification
Fungicide application
Biological controlBiological control
Weed and pest management
Agronomic measures
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PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
GOOD AGRICULTURAL PRACTICE (GAP)( )
Resistant varieties
Crop rotation
Soil treatmentSoil treatment
In‐planta detoxification
Fungicide application
Biological controlBiological control
Weed and pest management
Agronomic measures
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In‐planta detoxificationBioBio‐‐Degradation of Degradation of AflatoxinAflatoxin B1 by B1 by ActinomycetesActinomycetes
AFB1 degradation by cultures of Rhodococcus erythropolis ATCC 4277, Streptomyceslividans TK 24 and S. aureofaciens ATCC10762 after 24 h incubation at 30 °C and pH 6.0lividans TK 24 and S. aureofaciens ATCC10762 after 24 h incubation at 30 C and pH 6.0
Eshelli et al., 2015
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In‐planta detoxificationBiologicalBiological detoxificationdetoxification ofof deoxynivalenoldeoxynivalenol
De‐epoxydation: requires anaerobicditiconditions
Oxidation and epimerization: catalyzed by systems difficult toexpress and assembly in plantsexpress and assembly in plants
Use of intact microorganisms in feed additives
Mineralization: isolation of active pure cultures extremely difficult
Structure of deoxynivalenol (DON) with the designation of the ring system A‐C and targets for detoxification
Acetylation and glycosylation of C OH ll d t d d
pure cultures extremely difficult
C3‐OH: well understood and achievable in transgenic plants Molecular markers associated with genes
encoding plant glucosyltransferases that detoxify DON will help breeders develop new wheat
Karlovsky, 2011
DON will help breeders develop new wheat varieties resistant against FHB
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In planta detoxificationDeoxynivalenolDeoxynivalenol InactivatingInactivating UDPUDP GlucosyltransferaseGlucosyltransferase
In‐planta detoxificationDeoxynivalenolDeoxynivalenol‐‐InactivatingInactivating UDPUDP‐‐GlucosyltransferaseGlucosyltransferase
fromfrom barleybarley in in yeastyeast
Deoxynivalenol (DON) resistance phenotype Deoxynivalenol (DON)‐resistance phenotype conferred by expression of candidate barley UDP‐
glycosyltransferases (UGT) in yeastConversion of deoxynivalenol (DON) into DON‐3‐O‐glucoside (D3G) and release into the culture fluid by transformed yeasty y
Schweiger et al., 2010
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In‐planta detoxificationZearalenoneZearalenone (ZEN) is detoxified by ZHD101, a (ZEN) is detoxified by ZHD101, a lactonohydrolaselactonohydrolase from from ClonostachysClonostachys rosearoseayy yy
Degradation of ZEN by protein extracts from transgenic rice plants
Higa‐Nishiyama et al., 2005
Degradation of ZEN by protein extracts from transgenic rice plants transformed with zhd101
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PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
GOOD AGRICULTURAL PRACTICE (GAP)( )
Resistant varieties
Crop rotation
Soil treatmentSoil treatment
In‐planta detoxification
Fungicide application
Biological controlBiological control
Weed and pest management
Agronomic measures
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DON contamination in wheat plants untreated and treated with fungicideg
F i idF i id
FungicideFungicideUntreatedUntreated
Lower levels of
0.910.910 10 1
FungicideFungicide
0.060.06NdNd
0.0840.0840.0400.040
0.0400.040NdNd
UntreatedUntreatedLower levels ofDON in treated
plants
UntreatedUntreated FungicideFungicide
0.10.1 NdNd
6.66.6 4.44.4
FungicideFungicideUntreatedUntreated 14.6114.6116.0016.00
5.315.31NdNd
0,310,31 3,743,74 1.961.964.604.60
0,590,59NdNd
1-2 cm 2nd inter-node Milk ripening Vitreous ripening1 2 cm wheat head
2nd inter nodegrowth stage
Milk ripening Vitreous ripening
Moretti et al., 2014
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Effect of fungicide application on DONcontamination in wheat plantsp
S i t l 2015Scarpino et al., 2015
Fusarium diseaded kernelsFusarium diseaded kernelswith different fungicide
treatments
Mesterházy, 2014
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New products development…
New chemical classes of fungicides(e.g. SDHI and triazoles), more
efficient against FHB species, withlow toxicological risk for humanslow toxicological risk for humansand environement (Syngenta)
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PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
GOOD AGRICULTURAL PRACTICE (GAP)( )
Resistant varieties
Crop rotation
Soil treatment
EXPENSIVE
Soil treatment
In‐planta detoxificationEXPENSIVE
POTENTIAL HARM TO BENEFICIAL INSECTS AND ENVIRONMENT
Fungicide application
Biological controlRESIDUES IN FOOD
HAZARD FOR THE HEALTH AND SAFETY OF WORKERS HANDLING FUNGICIDES
Biological control
Weed and pest managementOF WORKERS HANDLING FUNGICIDES
Agronomic measures
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PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
GOOD AGRICULTURAL PRACTICE (GAP)( )
Resistant varieties
Crop rotation
Soil treatmentSoil treatment
In‐planta detoxification
Fungicide application
Biological controlBiological control
Weed and pest management
Agronomic measures
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Biological controlg
69‐80% AFLATOXINS CONTROL REDUCTION AFLASAFETM
i t f f t ig i IN MAIZE WITH NON‐TOXIGENIC TREATED WITH AFLASAFE™ IN NIGERIA
a mixture of four atoxigenicAspergillus flavus strains
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Biological control
Berries treated withAureobasidium
pullulans
Control berries infectedwith Aspergilluscarbonariuspullulans carbonarius
Sour Rot IncidenceAspergillus carbonarius
populations in grape berries Ochratoxin A content
Dimakopoulou et al., 2008
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Treatment of maize stubble residues with biocontrol agents exposed to fieldconditions to reduce the inoculum of toxigenic Fusarium species
MaizeMaize stalksstalks treatedtreated withwith::•• bacterialbacterial strainsstrains ofof PseudomonasPseudomonas•• fungalfungal strainsstrains ofof ClonostachysClonostachys rosearosea
PlacedPlaced onon soilsoil toto fieldfield conditionsconditions
Samples of mize stalks• untreated, • after 3 months and
AnalysedAnalysed byby TaqManTaqMan PCR PCR toto detectdetect::‐‐ FF. . graminearumgraminearum‐‐ FumonisinFumonisin producingproducing FusariumFusarium speciesspecies ((commonlycommonly
• after 6 months in field conditionsFumonisinFumonisin producingproducing FusariumFusarium speciesspecies ((commonlycommonlyF. F. verticillioidesverticillioides and and F. F. proliferatumproliferatum, in Italy), in Italy)
84% reduction after 3 months
88% reduction after 6 months
57% reduction after 3 months
82% reduction after 6 months
Clonostachys rosea can be considered a potentially useful antagonist in controlling fumonisins‐producing Fusarium species and F. graminearum
Environmental conditions have a strong influence on the capability of microbial antagonists to control the o o o o g o p y o o go o o odiseases in the field
Somma et al.
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PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
GOOD AGRICULTURAL PRACTICE (GAP)( )
Resistant varieties
Crop rotation
Soil treatmentSoil treatment
In‐planta detoxificationEXPENSIVE
UNRELIABLE (variation in the application due
Fungicide application
Biological control ( ppto environmental conditions)
LIMITED EFFICIENCY (survival)
Biological control
Weed and pest management
IMPACT ON ECOSYSTEM?Agronomic measures
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PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
GOOD AGRICULTURAL PRACTICE (GAP)( )
Resistant varieties
Crop rotation
Soil treatmentSoil treatment
In‐planta detoxification
Fungicide application
Biological controlBiological control
Weed and pest management
Agronomic measures
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Bt corn
Bt corn hybrids (left) can reduce mycotoxin contamination in corn compared to non Bt corn hybrids (right) corn compared to non‐Bt corn hybrids (right)
(Courtesy G. Munkvold)
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PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
PREPRE‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
GOOD AGRICULTURAL PRACTICE (GAP)( )
Resistant varieties
Crop rotation
Soil treatmentSoil treatment
In‐planta detoxification
Fungicide application
Biological controlBiological control
Weed and pest management
Agronomic measures
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Agronomic measuresEffect of the density of previous maize crop residues on the soil surface on FHB incidence and severity and DON contamination in wheat kernels
AMOUNT OF RESIDUES
Blandino et al., 2010
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Importance of different crop management practices for reducing the risks of mycotoxin contamination in maize
Fusarium ear rota Gibberella ear rotb Aspergillus ear rotc
Fumonisins DON, Zearalenone Aflatoxins
Hybrid selection ++ ++ ++
Insect control ++ + ++
Early planting ++ ++ +y p g
Irrigation management ++ + ++
Timely harvest ++ ++ ++
Crop rotation and tillage + ++ +
Plant Density + + +
Fertilization + + +Fertilization + + +
Weed management +/? ? +/?
Biological control ? ? +
Seed treatment ? ? ?
Fungicide application ? ? ?
++ = major effect; + = minor effect; ? = uncertain or insufficient evidence++ = major effect; + = minor effect; ? = uncertain or insufficient evidenceaCaused primarily by F. verticillioides, F. proliferatum, and F. subglutinansb Caused primarily by F. graminearum and F. culmorum (Gibberella zeae)a Caused primarily byA. flavus and A. parasiticus
Munkvold, 2014
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POSTPOST‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
POSTPOST‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
SortingSorting
Storageg
Detoxification
Intake prevention
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POSTPOST‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
POSTPOST‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
SortingSorting
Storageg
Detoxification
Intake prevention
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SortingVisual inspectionVisual inspection
CleaningCleaning
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Reduction of aflatoxins in maize by combining mechanical and optical sorting technologies
Total AFs reduction (%)( )
Separator – Aspirator – Sortex 65‐78
Separator – Aspirator – Concentrator ‐ Sortex 84‐76
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POSTPOST‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
POSTPOST‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
Sorting CONTAMINATION NOT DETECTABLESorting
StorageDISPOSAL OF REFUSES
g
Detoxification
Intake prevention
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What can farmers do with refuseunder low legal limits of
mycotoxin contamination?mycotoxin contamination?
Feed to livestock ?Burn for energy
Utilyze in a biogas facility
??
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POSTPOST‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
POSTPOST‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
SortingSorting
Storageg
Detoxification
Intake prevention
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Storage• aw for molds and yeast growth : 0.61 • lower limit for growth of mycotoxigenic molds: 0.78
Water activity (aWater activity (aww))
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Storage
GPRS
Sensor
Wireless sensor k d i
Sensor Node
network devices used into pilot scale grain silos t it i g
Sensor Node
to monitoring environmental parameters
Ambient intelligence system Sensor Node
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Storage
easy‐to‐use interface
Software for monitoring parameter in silosSoftware for monitoring parameter in silos
S d fS d fSensor node for:Sensor node for:•Temperature and Relative Humidity•Carbon Dioxide, Temperature and RH Sensor Node
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POSTPOST‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
POSTPOST‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
SortingSorting
Storageg
Detoxification
Intake prevention
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Mycotoxin‐detoxifying agents... additives for mycotoxin decontamination of feeds
BBIODEGRADATIONIODEGRADATIONMODE OF ACTION
AADSORPTIONDSORPTION
BBIOPROTECTIONIOPROTECTION
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Mycotoxin‐detoxifying agents
EFFICACY
... additives for mycotoxin decontamination of feeds
BBIODEGRADATIONIODEGRADATIONEFFICACY
AADSORPTIONDSORPTION
BBIOPROTECTIONIOPROTECTION
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DetoxificationChemical detoxification of Chemical detoxification of deoxynivalenoldeoxynivalenol
Paulick et al., 2015
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DetoxificationDegradation of Degradation of fumonisinfumonisin B1 by B1 by
cinnamon essential oilcinnamon essential oilcinnamon essential oilcinnamon essential oil
Xing et al., 2014
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POSTPOST‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINS
POSTPOST‐‐HARVEST CONTROL HARVEST CONTROL OF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINSOF MYCOTOXINS
SortingSorting
Storageg
Detoxification
Intake prevention
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Intake preventionPrediction models for Prediction models for FusariumFusarium head blight of wheat caused head blight of wheat caused
by by FusariumFusarium graminearumgraminearum
De Wolf and Paul, 2014
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Major challenges formycotoxin mitigation
• CONFLICT BETWEEN FOOD SAFETY AND FOOD SECURITYFOOD SECURITY
• PERCEPTION OF CHRONIC EFFECTS
• ACCESS TO SMALLHOLDER FAMILIES