Biopesticides, a review of active Biopesticides, a review of active agentsagents

Andy Cherry,Andy Cherry,Natural Resources Institute,Natural Resources Institute,University of Greenwich, UKUniversity of Greenwich, UK

Ghana, 27 Ghana, 27 –– 30 June 2005.30 June 2005.

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Active agentsActive agentsVirusesViruses

Baculoviruses Baculoviruses –– granuloviruses & nucleopolyhedrovirusesgranuloviruses & nucleopolyhedrovirusesCypovirusesCypoviruses

BacteriaBacteriaBacillus thuringiensis, B. Bacillus thuringiensis, B. spaericusspaericus, , PaenibacillusPaenibacillus popillaepopillae

FungiFungiMetarhizium, Beauveria, Trichoderma, Verticillium……Metarhizium, Beauveria, Trichoderma, Verticillium……

ProtozoaProtozoaNosema, Nosema, VairimorphaVairimorpha

NematodesNematodesSteinernema, HeterorhabditisSteinernema, Heterorhabditis

SemiochemicalsSemiochemicals

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Viruses infecting invertebratesViruses infecting invertebrates

DNA virusesDNA viruses–– Double stranded DNADouble stranded DNA

PoxviridaePoxviridae

IridoviridaeIridoviridae

BaculoviridaeBaculoviridae -- NPV and GVNPV and GV

PolydnaviridaePolydnaviridae

–– Single stranded DNASingle stranded DNA

ParvoviridaeParvoviridae

RNA virusesRNA viruses–– Double stranded RNADouble stranded RNA

RheoviridaeRheoviridae –– cypoviruscypovirusBirnaviridaeBirnaviridae

–– Single stranded RNA (Single stranded RNA (--))RhabdoviridaeRhabdoviridaeBunyaviridaeBunyaviridae

–– Single stranded RNA (+)Single stranded RNA (+)PicornaviridaePicornaviridae, , TogaviridaeTogaviridae, , TetraviridaeTetraviridae, , FlaviviridaeFlaviviridae, , NodaviridaeNodaviridae

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Entomopathogenic virusesEntomopathogenic viruses

BaculoviridaeBaculoviridae ((dsds DNA)DNA)–– NuclepolyhedrovirusNuclepolyhedrovirus

–– GranulovirusGranulovirus

ReoviridaeReoviridae ((dsds RNA)RNA)–– CypovirusCypovirus

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BaculovirusesBaculoviruses: Key reference: Key reference

OECD (2002) Consensus Document on Information used in the OECD (2002) Consensus Document on Information used in the Assessment of Environmental Applications involving Assessment of Environmental Applications involving BaculovirusBaculovirus. . Series on Harmonization of Regulatory Oversight in BiotechnologySeries on Harmonization of Regulatory Oversight in Biotechnology No. No. 20. OECD Environment, Health and Safety Publications. 20. OECD Environment, Health and Safety Publications. ENV/JM/MONO(2002)1. ENV/JM/MONO(2002)1.

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BaculovirusesBaculoviruses

2 microns

Spodoptera littoralis

From Hunter-Fujita et al

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Baculoviruses Baculoviruses --Mode of actionMode of action

From Hunter-Fujita et al

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BaculovirusesBaculoviruses

Found only in invertebratesFound only in invertebrates

No member of the family is known to infect plant or No member of the family is known to infect plant or vertebratevertebrate

Most have narrow host insect range, and infectivity is Most have narrow host insect range, and infectivity is restricted to the original host genus or familyrestricted to the original host genus or family

Susceptibility of Alternative HostsSusceptibility of Alternative Hosts

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BaculovirusesBaculoviruses

Toxicity test results from 1970s/80s of 29 Toxicity test results from 1970s/80s of 29 NPVsNPVsindicated no toxicity or pathogenicity. Doses were indicated no toxicity or pathogenicity. Doses were generally 10 generally 10 –– 100 x the “per acre” (1 acre = 0.45ha) 100 x the “per acre” (1 acre = 0.45ha) field rate equated to a 70kg person.field rate equated to a 70kg person.

HeliothisHeliothis zeazea NPV most extensively tested for toxicity in NPV most extensively tested for toxicity in humans and led to registration of “humans and led to registration of “ElcarElcar” by ” by SandozSandoz in in USA. USA.

Toxicity studies Toxicity studies -- mammalsmammals

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BaculovirusesBaculoviruses

Acute toxicityAcute toxicity--pathogenicity tests in mouse, rat, guinea pig, rabbit, pathogenicity tests in mouse, rat, guinea pig, rabbit, monkey and man at 6x10monkey and man at 6x1099 –– 3 x 103 x 101212 OB / kg.OB / kg.Skin irritation sensitivity tests in guinea pigs, rabbits and maSkin irritation sensitivity tests in guinea pigs, rabbits and man at 10n at 1066

and 10and 1077 OB / mmOB / mm2 2 skin.skin.Eye irritation tests in rabbits with 10Eye irritation tests in rabbits with 1055 and 2x10and 2x1066 OB / eyeOB / eyeSubacuteSubacute toxicitytoxicity--pathogenicity tests and subcutaneous injection into pathogenicity tests and subcutaneous injection into mice, rats, dogs and rhesus monkeys.mice, rats, dogs and rhesus monkeys.Teratogenicity and carcenogenicity studies in rats and mice at 1Teratogenicity and carcenogenicity studies in rats and mice at 10099 ––3.5x103.5x101212 OB / kg.OB / kg.

Toxicity studies Toxicity studies –– mammals cont.mammals cont.No effects of HzNPV found in:

Similar but less extensive results for many other Similar but less extensive results for many other NPVsNPVs from the from the 1970s/80s1970s/80s

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BaculovirusesBaculoviruses

BirdsBirdsAble to pass NPV through the alimentary tract unaffected Able to pass NPV through the alimentary tract unaffected No deleterious effectsNo deleterious effects

Aquatic organismsAquatic organismsNo adverse effectsNo adverse effects

Beneficial insectsBeneficial insectsNo direct effect on parasitoids, predators and pollinatorsNo direct effect on parasitoids, predators and pollinatorsIndirect effects on parasitoids resulting from host deathIndirect effects on parasitoids resulting from host death

Toxicity studies Toxicity studies –– wildlifewildlife

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BaculovirusesBaculoviruses

Toxicity tests designed for testing effects of chemicals on Toxicity tests designed for testing effects of chemicals on vertebrates are insufficientvertebrates are insufficient

Results reported in Results reported in GrGröönerner (1986) indicate no virus induced (1986) indicate no virus induced antibody production in test mammals and chicken.antibody production in test mammals and chicken.

No cytogenetic effects of baculoviruses in mammalian cells eitheNo cytogenetic effects of baculoviruses in mammalian cells either r in vivoin vivo or or in vitroin vitro..

Pathology studiesPathology studies

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BaculovirusesBaculoviruses

AcNPVAcNPV inoculated into vertebrate cells can be takeninoculated into vertebrate cells can be taken--up and the degree of upup and the degree of up--take depends on cell type, take depends on cell type, temperature, time and viral phenotype.temperature, time and viral phenotype.

BUT, none of the human and nonhuman vertebrate BUT, none of the human and nonhuman vertebrate lines tested showed evidence of viral replication. lines tested showed evidence of viral replication.

NPVsNPVs unable to activate retroviruses in mammalian unable to activate retroviruses in mammalian cell linescell lines

VirusVirus--cell interactions in vitrocell interactions in vitro

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Baculovirus registrationBaculovirus registration

The most comprehensive / developed registration The most comprehensive / developed registration guidelines come from the US EPA:guidelines come from the US EPA:

http://http://www.epa.gov/pesticides/biopesticides/ai/viruses.htmwww.epa.gov/pesticides/biopesticides/ai/viruses.htm

European regulations well developed but complicated European regulations well developed but complicated by the number of countries, despite the efforts at by the number of countries, despite the efforts at harmonising across the EC.harmonising across the EC.

(EC directive 91/414/EEC)(EC directive 91/414/EEC)

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BaculovirusesBaculoviruses

a list of the baculoviruses regulated as pesticide a list of the baculoviruses regulated as pesticide active ingredients by the US EPA Office of Pesticide active ingredients by the US EPA Office of Pesticide Programs as of May 2005Programs as of May 2005

Anagrapha falcifera NPVCydia pomonella GV Douglas fir tussock moth NPV Gypsy moth NPV Helicoverpa zea NPV Indian meal moth GVMamestra configurata NPV (pending) Spodoptera exigua NPV

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Baculoviruses Baculoviruses –– US EPA fact sheetUS EPA fact sheetIII. ASSESSING RISKS TO HUMAN HEALTH

These viruses infect only the target insect larvae and closely related species. Toxicity tests show that the viruses pose no risk to the public. Workers wear protective clothing to prevent possible irritation from handling and applying the product.

IV. ASSESSING RISKS TO THE ENVIRONMENT

Tests show that the GV and NPVs that EPA has registered as pesticide active ingredients specifically infect only certain species of moth larvae. The viruses do not harm other organisms, including plants, beneficial insects, other wildlife, or the environment. These viruses occur naturally in their insect hosts.

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CypovirusesCypoviruses: Mode of action: Mode of action

PolyhedraPolyhedra ingested and dissolved in larval ingested and dissolved in larval midgutmidgut

VirionsVirions released and attach to released and attach to midgutmidgut columnar cellscolumnar cells

Viral core enters cell cytoplasmViral core enters cell cytoplasm

RNA transcription and replicationRNA transcription and replication

RNA occluded in capsulesRNA occluded in capsules

Virus capsules occluded by Virus capsules occluded by virogenicvirogenic stromastroma to form occlusion to form occlusion

bodiesbodies

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CypovirusesCypoviruses ((RheoviridaeRheoviridae))

No CPV has been found infecting vertebrates or plants No CPV has been found infecting vertebrates or plants ((BelloncikBelloncik, 1989), 1989)

Dendrolimus spectabilis CPV registered in Japan in 1974. Safety test results generally negative.

– Katagiri, K. (1981) Pest control by cytoplasmic polyhedrosos viruses. In: Microbial control of pests and plant diseases 1970-1980. (Ed Burges, H.D.) Academic Press.

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Bacterial EntomopathogensBacterial EntomopathogensSporeSpore--forming Bacteriaforming Bacteria

Paenibacillus (=Bacillus popillaePaenibacillus (=Bacillus popillae))Bacillus sphaericus Bacillus sphaericus Bacillus thuringiensisBacillus thuringiensis–– Related mammalian pathogens:Related mammalian pathogens:

Bacillus cereusBacillus cereusBacillus Bacillus anthracisanthracis

NonNon--spore forming spore forming sppspp..Serratia entomophilaSerratia entomophilaClostridium Clostridium bifermentansbifermentans

TransgenicsTransgenicsPseudomonas Pseudomonas sppspp..

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Bacillus popilliaeBacillus popilliae

Causes “milky disease” in scarab larvaeCauses “milky disease” in scarab larvaeHostHost--specific, apparently an obligate pathogen on scarabeid beetlesspecific, apparently an obligate pathogen on scarabeid beetlesUsed for many years in control of scarab larvaeUsed for many years in control of scarab larvaeSpores can last in the environment for several years. Spores can last in the environment for several years. Ability to recycleAbility to recycle

Mode of actionMode of actionChronic Chronic –– nonnon--toxic mode of actiontoxic mode of actionCause of death may be infection and physiological starvationCause of death may be infection and physiological starvationConcentrations of bacteria in beetle haemolymph can be 3*10Concentrations of bacteria in beetle haemolymph can be 3*1099

bacteria/ml bacteria/ml

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B. popillaeB. popillae safetysafety

Highly specific, nonHighly specific, non--toxic mode of action toxic mode of action -- no no toxicity known to mammals, birds, or fish. toxicity known to mammals, birds, or fish. Does not grow above 35.5 Does not grow above 35.5 ooCC so can not persist in so can not persist in vertebratesvertebratesNonNon--target effects: None knowntarget effects: None known

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Bacillus sphaericus

Activity restricted to certain Diptera

Effective against mosquitoes

Ability to recycle to produce new infections

Spore and multiple toxin crystals

Lasts up to nine months in aquatic environments

Better than Bt israelensis in polluted waters

No measurable health effects or environmental

risks

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Bacillus thuringiensisBacillus thuringiensis

Foliar applications have a typical persistence of 4Foliar applications have a typical persistence of 4--5 days. 5 days. Spores are viable for years in the soil (not exposed)Spores are viable for years in the soil (not exposed)Rarely recycles in natureRarely recycles in nature

Uses:Uses:field cropsfield cropsforest pestsforest pestsvector control in aquatic ecosystems vector control in aquatic ecosystems

Number one biopesticide worldNumber one biopesticide world--widewide

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Bt varietiesBt varieties

kurstakikurstaki –– standard lepidopteran strainstandard lepidopteran strain

aizawaiaizawai –– lepidopteran strainlepidopteran strain

israelensisisraelensis –– diptera (mosquito, blackflies) diptera (mosquito, blackflies)

tenebrionistenebrionis –– some beetles (e.g.Colorado potato beetle)some beetles (e.g.Colorado potato beetle)

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Bt Mode of ActionBt Mode of Action

Kills by direct toxicity, not infectionKills by direct toxicity, not infection

Parasporal bodies = occlusion bodies = crystals containing toxinParasporal bodies = occlusion bodies = crystals containing toxins.s.

Crystal toxins = Crystal toxins = δδ ((delta)delta)--endotoxinsendotoxins

Other toxins:Other toxins:

HaemolysinsHaemolysins

ββ--exotoxinsexotoxins

enterotoxinsenterotoxins

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Delta EndotoxinsDelta Endotoxins

Activation of endotoxins– monomeric protoxin to active δδ--endotoxins endotoxins

Mode of Action– active δδ--endotoxins bind with endotoxins bind with gut receptors – creates pores interfering with ion transport– results in gut paralysis– high doses lead to gut lysis and rapid death

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2h after distilled water

3h after Bt endotoxin

Effect of Bt endotoxin on insect gut tissue

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Beta ExotoxinsBeta Exotoxins

ProteinaceousProteinaceous, sometimes excreted in the growth media. , sometimes excreted in the growth media.

Detected in several strains from several subDetected in several strains from several sub--sppspp..

Inhibits biosynthesis of RNAInhibits biosynthesis of RNA

Broadly toxic to vertebrates Broadly toxic to vertebrates

Not associated with the crystal proteins. Not associated with the crystal proteins.

Are genetically associated with crystal proteins (on same Are genetically associated with crystal proteins (on same plasmid)plasmid)

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EnterotoxinsEnterotoxins

Known from the closely related Known from the closely related Bacillus cereusBacillus cereus. Some . Some varieties of varieties of B. cereusB. cereus cause gastroenteritis (food cause gastroenteritis (food poisoning). poisoning).

Some commercially used strains of Bt can produce Some commercially used strains of Bt can produce enterotoxins. No conclusive case of Btenterotoxins. No conclusive case of Bt--induced induced gastroenteritis gastroenteritis

Bt enterotoxin ten times less than that produced by Bt enterotoxin ten times less than that produced by Bacillus cereusBacillus cereus..

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Safety of BtSafety of Bt

Humans Humans –– After 60 years of use, clinical After 60 years of use, clinical infections sufficiently rare that they are infections sufficiently rare that they are printed in medical journals. printed in medical journals. Human testing showed no undesirable effects Human testing showed no undesirable effects –– ingestion (3ingestion (3--10 * 1010 * 1099 spores/day for 3spores/day for 3--5 days)5 days)–– inhalation (100mg for five days) inhalation (100mg for five days)

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MammalsMammals

OralOralNo undesirable effects ever reported No undesirable effects ever reported Bt remains confined to the gastrointestinal tract Bt remains confined to the gastrointestinal tract (McClintok, 1995) (McClintok, 1995)

Acute PulmonaryAcute Pulmonary2.6*102.6*1077 spores/kg. spores/kg. Clearance from lung with no adverse effects.Clearance from lung with no adverse effects.

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MammalsMammals

IntravenousIntravenousBt israelensiBt israelensis declines steadily in the bloodstream, s declines steadily in the bloodstream, lungs, kidneys, brain. lungs, kidneys, brain. Indicative of inability to replicate and/or persistIndicative of inability to replicate and/or persist

IntraperitonealIntraperitonealNo effects in mice at doses of 10No effects in mice at doses of 1066 or 10or 1077 spores spores At 10At 1088 spores, substantial mortality appears (as also spores, substantial mortality appears (as also happens with “harmless” bacteria)happens with “harmless” bacteria)

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MammalsMammals

Dermal/OcularDermal/OcularSlight to moderate skin irritation has been documented. Slight to moderate skin irritation has been documented. Reversible in 7 daysReversible in 7 daysCornea undamaged. Cornea undamaged. Formulation, not Bt. was assumed to be the cause.Formulation, not Bt. was assumed to be the cause.

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BirdsBirds

No undesirable effects in birds from feeding wild No undesirable effects in birds from feeding wild birds or chickens with spores or with infected birds or chickens with spores or with infected insectsinsects

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Fungi as biocontrol agents for plants, Fungi as biocontrol agents for plants, plant diseases and arthropodsplant diseases and arthropods

Potential safety issues:Potential safety issues:Competitive displacement of nonCompetitive displacement of non--target organismstarget organismsAllergenicityAllergenicityToxigenicityToxigenicity to NTOsto NTOsPathogenicity to NTOsPathogenicity to NTOs

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Competitive displacement:Competitive displacement:Competitive occupation of nichesCompetitive occupation of niches

Intended / unintendedIntended / unintended

AllergenicityAllergenicitySpores can cause allergic reactionsSpores can cause allergic reactionsReports of allergic reactions among mass production Reports of allergic reactions among mass production workersworkersFungal Fungal bcabca are not among the species responsible for are not among the species responsible for production of common allergiesproduction of common allergies

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ToxicityToxicityFungi secrete wide array of compounds with biological Fungi secrete wide array of compounds with biological activity against other organismsactivity against other organismsFunction differs depending on the ecological niche of the Function differs depending on the ecological niche of the fungus.fungus.May play a role in pathogenesisMay play a role in pathogenesis--antagonism in antagonism in bcabca

MetarhiziumMetarhizium anisopliae anisopliae –– destruxinsdestruxins and and cytochalasinscytochalasinsBeauveriaBeauveria bassiana bassiana –– bassianinbassianin, , beauvericinbeauvericin, , beauveriolidebeauveriolide, , bassianolidebassianolideBeauveriaBeauveria brogniatiibrogniatii -- oosporeinoosporeinHirsutellaHirsutella thompsoniithompsonii –– hirsutellinhirsutellinTrichodermaTrichoderma harzianumharzianum –– peptaibolspeptaibolsFusariumFusarium sppspp. . –– trichothecenestrichothecenes, , beauvericinbeauvericinColletotrichumColletotrichum -- colletotrichincolletotrichin

Toxin production varies according genus and isolateToxin production varies according genus and isolate

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Toxins Toxins –– a reason for exclusion?a reason for exclusion?

Presence of toxins should not necessarily preclude use of BCA inPresence of toxins should not necessarily preclude use of BCA inbiocontrolbiocontrol

Considerations:Considerations:Presence of toxin in formulated product?Presence of toxin in formulated product?Fate of toxin after application?Fate of toxin after application?Accumulation of toxin in host / environment?Accumulation of toxin in host / environment?

No documented accounts of toxin levels rising No documented accounts of toxin levels rising after epizootic/epidemicsafter epizootic/epidemicsNo reports of No reports of bcabca metabolites entering the metabolites entering the food chainfood chainRisks are very lowRisks are very low

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RAFBCARAFBCA((RRisk isk AAssessment of ssessment of FFungal ungal BBiological iological CControl ontrol AAgents)gents)

http://www.rafbca.com

EU project that has generated:EU project that has generated:

1.1. Data on fungal metabolitesData on fungal metabolites2.2. Tools for risk assessmentTools for risk assessment

in order to accelerate registration of fungal in order to accelerate registration of fungal bcabca in Europein Europe

Major conclusion of RAFBCA is that metabolites of selected fungaMajor conclusion of RAFBCA is that metabolites of selected fungal BCA do l BCA do not enter the food chain, and therefore do not pose a risk to conot enter the food chain, and therefore do not pose a risk to consumer and nsumer and animal health. animal health. Risk assessment of each metabolite produced by a microbe would bRisk assessment of each metabolite produced by a microbe would be very e very expensive and certainly deter most small and medium sized enterpexpensive and certainly deter most small and medium sized enterprisesrises

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PathogenicityPathogenicity

Pathogenicity towards host is desired effectPathogenicity towards host is desired effectAbility to infect is usually constrained within a host groupAbility to infect is usually constrained within a host group

But, pathogenicity towards NTOs, including vertebrates, But, pathogenicity towards NTOs, including vertebrates, is reported:is reported:

–– Reports include:Reports include:B. bassiana B. bassiana infecting alligators, giant tortoises, inland silverside fish infecting alligators, giant tortoises, inland silverside fish embryos, grass shrimpsembryos, grass shrimpsM. anisopliaeM. anisopliae infecting inland silverside fish embryos, infecting inland silverside fish embryos, immunocompentent and immunoincompetent humansimmunocompentent and immunoincompetent humansPaecilomyces lilacinusPaecilomyces lilacinus in immunocompentent and in immunocompentent and immunoincompetent humansimmunoincompetent humansConidiobolusConidiobolus coronatuscoronatus associated with lesions in horses and associated with lesions in horses and humanshumans

Photo: G. Goergen

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Entomopathogenic microsporidiaEntomopathogenic microsporidia

Microsporidia are genetically aligned with fungiMicrosporidia are genetically aligned with fungiPrincipal genusPrincipal genus = Nosema= Nosema. . Nosema locustae Nosema locustae is the only sp. Registered by US EPA is the only sp. Registered by US EPA as biopesticide.as biopesticide.–– Used in locust control for > 20 yrs. Used in locust control for > 20 yrs. –– Conventional and maximum challenge testing gave no illness or Conventional and maximum challenge testing gave no illness or

infection. infection.

Weight of evidence is against infection in mammals.Weight of evidence is against infection in mammals.–– Reproduction in some mammalian cell lines at <37Reproduction in some mammalian cell lines at <37°°C.C.

No effect on physical environmentNo effect on physical environmentSafety concerns relate only to effect on NTOsSafety concerns relate only to effect on NTOs

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NonNon--target effects of microsporidiatarget effects of microsporidia

Same as for fungi Same as for fungi –––– Competitive displacementCompetitive displacement–– AllergenicityAllergenicity–– PathogenicityPathogenicity–– Toxicity of metabolites and antibioticsToxicity of metabolites and antibiotics

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Entomopathogenic nematodesEntomopathogenic nematodes

GeneraGenera = Steinernema = Steinernema and and HeterorhabditisHeterorhabditisSymbiotic relationship with specific bacteria spp in Symbiotic relationship with specific bacteria spp in XenorhabdusXenorhabdus andand Photorhabdus Photorhabdus respectivelyrespectivelyWide host rangeWide host rangeHighly effectiveHighly effectivePersistent and can recyclePersistent and can recycleMobileMobileLarge scale mass production possibleLarge scale mass production possible

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EPN effects on nonEPN effects on non--target hoststarget hosts

No ecological problems ever recordedNo ecological problems ever recordedConsidered safe for user and environment Considered safe for user and environment

Vertebrates:Vertebrates:No detrimental effects for mammals or plantsNo detrimental effects for mammals or plants

No evidence of infection in humans of nematodes or associated No evidence of infection in humans of nematodes or associated bacteriabacteriaSlight risk of allergic effects from nematodeSlight risk of allergic effects from nematode--bacterium materialbacterium materialRemote risk of toxicity from contaminating bacteriaRemote risk of toxicity from contaminating bacteria

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EPN nonEPN non--target effectstarget effectsArthropods:Arthropods:

Risk for NTOs with soilRisk for NTOs with soil--dwelling stage is greatestdwelling stage is greatest

Soil arthropods can be infected if exposed to high concentrationSoil arthropods can be infected if exposed to high concentrations but s but effects not long lasting.effects not long lasting.

May affect immature parasitoids directly / indirectlyMay affect immature parasitoids directly / indirectly

Some evidence for infection of predators feeding on infected hosSome evidence for infection of predators feeding on infected hosts ts (mostly in laboratory)(mostly in laboratory)

Honey bees susceptible if directly sprayed but colony not adversHoney bees susceptible if directly sprayed but colony not adversely ely affectedaffected

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EPN nonEPN non--target effectstarget effectsOECDOECD--COST expert group identified risk for invertebrate COST expert group identified risk for invertebrate NTO to be slight in treated fields. Data gathered to date NTO to be slight in treated fields. Data gathered to date indicate nematodes currently used in biocontrol pose few indicate nematodes currently used in biocontrol pose few problems for natural populations of invertebrate NTOs.problems for natural populations of invertebrate NTOs.

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Recommendations Recommendations (from OECD(from OECD--COST expert group)COST expert group)

Indigenous EPN should not be regulatedIndigenous EPN should not be regulated

Release of exotic EPN should be regulated as macroRelease of exotic EPN should be regulated as macro--organismsorganisms

Regulation should be at species levelRegulation should be at species level

Future regulations may require tailoring with regard to Future regulations may require tailoring with regard to EPNEPN

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SemiochemicalsSemiochemicals

AllelochemicalsAllelochemicalsAllomonesAllomonesKairomonesKairomonesSynomonesSynomones

PheromonesPheromones

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SemiochemicalsSemiochemicalsNonNon--toxic toxic –– modify behaviourmodify behaviourHighly specificHighly specificNatural occurrenceNatural occurrenceEffective at very low ratesEffective at very low rates

Comparable to natural emissionsComparable to natural emissionsHumans and NTO have very low direct Humans and NTO have very low direct exposureexposure

Dissipate rapidlyDissipate rapidlyRapidly degradedRapidly degradedCrop residues extremely lowCrop residues extremely low

Risk of adverse effects minimisedRisk of adverse effects minimisedNo reports of adverse effects No reports of adverse effects

Thanks for your attentionThanks for your attention