Features of a new biofungicide undergoing EU Registration...Honey bees Apis mellifera Dietary effect...
Transcript of Features of a new biofungicide undergoing EU Registration...Honey bees Apis mellifera Dietary effect...
ACTINOVATEFeatures of a new biofungicide undergoing
EU Registration
Carolina Fernández1, Ángela Sarro1, José Manuel Lara1 & Matthew Kowalski2
6th Annual Biocontrol Industry Meeting (ABIM) Lucerne, Switzerland October 24‐26, 2011
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CONTENTS
PART 1: BIOFUNGICIDE ACTINOVATE
Active ingredientBiological properties and risk assessmentCharacteristics of SP formulation
PART 2: SUITABILITY FOR IPM
Side effects on non‐target organismsEfficacy against fungal and bacterial diseasesCompatibility with PPPs
HISTORY: Streptomyces lydicusWYEC 108 (ACTINOVATE®)
Discovery: 1991Isolation from rhizosphere from linseed plant roots (Crawford et al., 1993)Screening of new actinomycetes to control damping‐off (Phytium ultimum)Selection of WYEC 108: Stable prolific sporulator (15‐35 ºC / pH 4.0‐8.0)
Patenting & Licensing1992: IRF (University of Idaho): plant growth / yield enhancer1995: US patent of WYEC 108 to control plant pathogens1998: worldwide licensing to Natural Industries (US)
Registration2004: USA (EPA 73314‐1) 2007: Canada (PMRA PRD2007‐10)2005‐2011: Turkey, Vietnam, Ecuador, Trinidad (in progress: Australia, Taiwan, Thailand, Costa Rica)2010: EU (RMS: The Netherlands)2011: Completeness Check Decision 2011/253/EU of 26 April 2011
PART I – BIOFUNGICIDE ACTINOVATE
ACTIVE INGREDIENT
Taxonomy Streptomyces lydicus strain WYEC 108Family: Noctuoidea; Orden: Actinomycetales; Class: Actinobacteria; Phylla: Actinobacteria Kingdom: Bacteria
Origin Rhizosphere linseed plant (South Downs, UK)
Distribution Widely distributed in most agricultural soils worldwide
Deposited ATCC 55445
Target Root damping‐off and foliar fungal pathogens
Growing Te Typical mesophylic specie (15‐35ºC; optimum 30ºC)
Growing pH 4.0 – 8.0
PART I – BIOFUNGICIDE ACTINOVATE
MODE OF ACTION
Competition For the space and nutrients.
Mycoparasitism The bacteria breaks down the cell walls of pathogenic fungus mycelium by excreting lytic enzymes (chitinases, glucanases, peroxidases)
Antibiosis Production and release of antimicrobial secondary metabolites
PART I – BIOFUNGICIDE ACTINOVATE
WYEC 108
Fusarium
Fusarium
High siderophore production: Improve mineral & nutrient uptake:a) Promotes plant growth even in the absence of pathogenb) Plant more vigorous /able to survive stressful conditions
Bactericide:Firelight (Erwinia amylovora), Citrus Canker (Xanthamonas axonopodis), Walnut Blight (Xanthomonas arboricola), Rice Leaf Blight (Xanthomonas oryzae), Bacterial spot (Xanthamonas perforans)
ADDITIONAL BENEFITS
PART I – BIOFUNGICIDE ACTINOVATE
ErwiniaXanthomonas
TOXICOLOGY: Acute Mammalian Toxicity
Guideline Study Toxicity Category
Results
152‐30OPPTS 870.1100
Acute Oral Toxicity
IV 5 male + 5 female rats (Sprague‐Dawley)No rats died. LD50 > 5.000 mg/kg
152‐32OPPTS 885.3150
Acute PulmonaryToxicity /Pathogen.
N/A S. lydicus was not toxic, infective, or pathogenic (rats). Clearance: liver‐lymph nodes 7d, kidneys 14d, lungs 28d
152‐33OPPTS 885.3200
Acute InjectionToxicity / Pathogenicity
N/A S. lydicus was not toxic, infective, or pathogenic (rats). Clearance: blood‐kidney‐lymph nodes 14d, lungs 21d,
liver‐spleen 28d
152‐34OPPTS 870.2500
Acute Dermal Irritation Study
IV 3 rabbits dosed (500 mg)Very slight erythema in 1 rabbitt with clearance by 24h
152‐35OPPTS 870.2400
Acute Eye Irritation Study
IV 3 rabbits dosed (0.1mL)Conjunctival irritation was reduced by 24 h
OECD 471, 1997 Reverse Mutation Assay(Ames test)
No mutagen
Salmonella typhimurium No genotoxic in vitro; Irrelevant to mammals
PART I – BIOFUNGICIDE ACTINOVATE
Active Ingredient Streptomyces lydicus strain WYEC 108Concentration 1x107 cfu/g (0.0371% TGAI)Physical State Solid powder (SP)Colour White palepH (CIPAC MT 75) 6.65 (1% p/v)Stability (Tª) 1 year (35ºC)
FORMULATION CHARACTERISTICS
PART I – BIOFUNGICIDE ACTINOVATE
1,00E+001,00E+011,00E+021,00E+031,00E+041,00E+051,00E+061,00E+071,00E+081,00E+09
0 50 100 150 200 250 300 350DAYS
VIAB
ILITY (CFU
/mL)
4ºC RT
SOIL DISEASE FOLIAR DISEASE
Fusarium LaveillulaPythium SpahaerotecaRhizoctonia ErisiphePhytophthora OidiumSclerotinia VenturiaArmillaria Botrytis
Alternaria
TARGET PATHOGENS
Fusarium Fusarium +ACTINOVATE
Rhizoctonia Rhizoctonia +ACTINOVATE
Phytoph. Phytoph. +ACTINOVATE
PART I – BIOFUNGICIDE ACTINOVATE
Pythium Pythium +ACTINOVATE
Guideline Group Target Test Remarks
OPPTS 885.4380
Terrestrian Organisms
Invertebrate:Honey bees Apis mellifera
Dietary effect No riskNOEC > 17980 µg/mL (≈ x20 commercial dose)
OPPTS 885.4050
Vertebrate:Nothern BobwhiteColinus virginatus
Avian Oral Pathog./Toxicity Test
No risk(1.25x108 cfu/kg body for 5d)
OPPTS 885.4300
Non‐Target Plants ‐ Efficacy trials ‐ Databases ‐ Background levels
Absence of adversed effects
OPPTS 850.1300 y 885.4240
AcuaticOrganisms
Invertebrate: Daphnia magna
Chronic Toxicity(21‐d static renewal test)
NOEC (survival) = 3.1x103cfu/mLNOEC (reprod.) = 1.6x103cfu/mL
OPPTS 850.1075
Vertebrate:Rainbow TroutOncorhynchus myskiss
Acute Toxicity Test(96‐h static test)
No riskLC50 > 1x106 cfu/mLNOEC > 1x106 cfu/mL
PART II – SUITABILITY FOR IPM
EFFECT ON NON‐TARGET ORGANISMS
‐ Potential Ecological Risk Minimal
PART II – SUITABILITY FOR IPM
DIRECTION FOR USE
Crop Soil Foliar
Vegetables 300 ‐ 500 g/Ha 250 ‐ 700 g / Ha
Seed coating 200 – 500 g / 300 ‐ 1200 mL(per 50 g seeds)
Turf 3 kg/Ha (initially)1 kg/Ha (every 6‐8 wks)
Cut flowers, bulbs and pot
plants
100‐200 g/500 L500 g/250 L (woody plants)
200‐300 g/50 kg bulbs250 ‐ 700 g / Ha
Petunia and Geranium (Reddy, 2007, Alabama, US)
PART II – SUITABILITY FOR IPM
EFFICACY TRIAL: Phytium and Rhizoctonia
Geranium ‐ Rhizoctonia
‐Root‐rot severity was reduced by ACTINOVATE ≈ same level than Chemical Standard‐Vigor was increased by ACTINOVATE (PGPR)
Petunia ‐ Pythium
0
0,51
1,5
22,5
3
3,5
44,5
5
Vigor Root‐rot Severity
Scale 0‐5 Untreated Control
Pathogen ControlACTINOVATEChemical Standard
0
0,51
1,5
22,5
3
3,5
44,5
5
Vigor Root‐rot Severity
Scale 0‐5 Untreated Cont
Pathogen ContACTINOVATEChemical Stand
‐ Preventive teatment 7 days before transplant and inoculation with pathogen‐ Damping‐off was evaluated 45 days after transplanting.
Potato crop (Chapingo, México)
EFFICACY TRIAL: Rhizoctonia solani
PART II – SUITABILITY FOR IPM
‐Dose‐response effect high dose = best results
0
10
20
30
40
50
60
70
80
90
100
36 DAT 71 DAT 110 DAT
Pathogen ControlACTINOVATE (1 kg/Ha)ACTINOVATE (1.5 kg/Ha)ACTINOVATE (2 kg/Ha)
67%77%
82%
65%
76%80%
61%
75%79%
Disease Incidence (%)
0
5
10
15
20
25
30
35
40
45
50
36 DAT 71 DAT 110 DAT
Contr
ACTIN
ACTIN
ACTIN
76%81%
85%
72%80%82% 79%
70%
81%
Disease Severity (%)
‐ ACTINOVATE preventively applied to potato seeds
Turf vr. Raleigh (Schlesselman & Bruno, 2006, Wharton, Texas, US)
PART II – SUITABILITY FOR IPM
EFFICACY TRIAL: Rhizoctonia solani
0
5
10
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20
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35
40
45
CONTROL Chemical 1 ACTINOVATE Chemical 2 Chemical 3
‐ ACTINOVATE efficacy was similar than efficacy showed by chemical standards
Disease Incidence (%)
‐ Two spray applications
PART II – SUITABILITY FOR IPM
EFFICACY TRIAL: Fusarium oxysporium fsp cyclaminis (FOC)
*Published: Crop Protection
0
20
40
60
80
100
120
AUDPC Dry Weight %VD
FOCT (1DAT) + ACT (7DAT) + FOC (14 DAT)F (1DAT) + ACT (7DAT) + FOC (14 DAT)
‐ Applying ACTINOVATE after a preventive fungicide showed promise supression of FOC
Trial I
AUDPC: Area Under the Disease Progressive Curve ; %VD: % Vascular DiscolorationT: Thiophanate methyl; F: Fludioxonil;
Cyclamen (*Elmer & McGovern, 2004, Gainesville, Florida US)
PART II – SUITABILITY FOR IPM
EFFICACY TRIAL: Fusarium oxysporium fsp cyclaminis (FOC)
*Published: Crop Protection
Trial II
‐ACTINOVATE rotated with a chemical fungicide showed a better suppresion of FOC than the chemical alone
AUDPC: Area Under the Disease Progressive Curve ; %VD: % Vascular DiscolorationF: Fludioxonil; Az: Azoxystrobin
Cyclamen (*Elmer & McGovern, 2004, Gainesville, Florida US)
0
20
40
60
80
100
120
AUDPC Dry Weight %VD
FOCFOC (1 DAT) + AZ‐ACT (7DAT) + ACT (14 DAT)FOC (1 DAT) + AZ (7DAT) + AZ (14 DAT)FOC (1 DAT) + F ‐ ACT (7DAT) + ACT (14 DAT)FOC (1 DAT) + F (7DAT) + F (14 DAT)
Trial II
COMPATIBILITY WITH PPPs
PART II – SUITABILITY FOR IPM
0102030405060708090
100
Triadimen
ol25
%
Copp
eroxychloride
Miclobu
tanil
24%
Ciprocon
azol
Methyl
Tiofan
ate
Procim
idon
e50
%
Citrus
extract 4
0%
potassium
phosph
ite
Copp
eroxychloride
Cymoxan
ile60
%
COMPA
TIBILITY
(%)
0102030405060708090
100
Bupirim
ate
2,5%
Kasu
gamyc
in
Dimetho
mor
ph15
%
Wettable
Sulphu
r
Thira
m 80%
COMPA
TIBILITY
(%)
Fungicide FungicideFungicide‐Bactericide Fungicide‐Bactericide
COMPATIBLE with ACTINOVATE NON‐COMPATIBLE with ACTINOVATE
‐Ten of them affected less than 80% the viability of ACTINOVATE spores.
CLOSING REMARKS
ACTINOVATE (Streptomyces lydicus strain WYEC 108)
• Efficient broad range biofungicide (bactericide)
• Plant Growth Promoter even in the absence of pathogen
• IPM tool (low risk substance compatible with other PPPs)
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