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

1 2

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


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


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


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


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


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


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


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)


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


‐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)


EFFICACY TRIAL: Rhizoctonia solani

0

5

10

15

20

25

30

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


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)


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


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)

Thank youwww.futurecobioscience.com

Features of a new biofungicide undergoing EU Registration...Honey bees Apis mellifera Dietary effect...

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