Why biopesticides?

(US EPA view)

Agents are: usually inherently less toxic than conventional chemical pesticides, relatively specific, decompose quickly, avoiding pollution, [compatible with] IPM programmes To use biopesticides effectively users need to know a great deal about managing pests.
Introduction_Biopesticides v.1.1

Introduction to Biopesticides
Biological & Chemical Control Methods 2009

http://www.epa.gov/pesticides/biopesticides/whatarebiopesticides.htm

Products (actual and potential)

Microbial control agents (MCA) Parasitoids, predators Biorationals (plantextracts, fermentation products etc.) Pheromones Genes for GM crops

Market taking off (at least in EU)

Almeria, southern Spain: protected vegetables production industry. 2006 - early 2007, high pesticide (e.g. OP) residue levels exceeding MRL in sweet peppers

(2009)

Increase in land area using biological control agents in sweet pepper crops in Almeria, Spain
impacted severely on sales of produce in German supermarket clients Protected environment
7000 6000 5000 Area (ha) 4000 3000 2000 1000 0 2003 2004 2005 Year 2006 2007

Working definition: living biopesticides

Biopesticides here applied strictly to living micro-organisms that: are specific as individual products and thus confer some environmental advantage (in contrast to most but not all chemicals), and have a limited period of activity - and usually are used with normal pesticide application techniques (unlike most classical biological control agents).

Microbial control agents (MCA)

Examples
Bacteria
Bacillus thuringiensis (live) B. subtilis:

Registered biopesticide products in the UK (July 2009)

Active Substance Bacillus subtilis QST713 Bacillus thuringiensis var kurstaki Beauveria bassiana ATC74040 Candida oleophila Strain O Coniothyrium minitans Strain CON/M/91-08 Cydia pomonella Granulosis Virus Cydia pomonella Granulosis Virus Lecanicillium muscarium (formerly Verticillium lecanii) Lecanicillium muscarium (formerly Verticillium lecanii) Pseudomonas chlororaphis MA 342 Zucchini Yellow Mosaic Virus Weak Strain Product Name Serenade ASO DiPel DF Naturalis-L Nexy1 Contans WG Cyd-X Cyd-X Extra Mycotal Vertalec Cerall Curbit Type of product Bacteria Bacteria Fungus Yeast Fungus Granulosis virus Granulosis virus Fungus Fungus Bacteria Plant virus Target(s) Botrytis/Grey mould Lepidoptera pests Whitefly, thrips Post harvest diseases Sclerotinia Codling moth Codling moth Whitefly, thrips, scale insects, mealybug Aphids Seed treatment for cereal diseases Plant viruses

Viruses
Cydia pomonella Granulosis Virus

Fungi
Metarhizium spp, Beauveria bassiana Lecanicillium muscarium (= Verticillium lecanii) Trichoderma spp.

Protozoa
Nosema locustae

Nematodes
Steinernema spp, Phasmarhabditis hermaphrodita

Exempted biopesticide products in the UK (July 2009)

Active Substance Steinernema feltiae Steinernema kraussei Heterorhabditis megidis Heterorhabditis megidis Heterorhabditis megidis Steinernema carpocasae Product Name Nemasys Nemasys L Nemasys H Lavanem Nemasys H Nemasys C Type of product Entomopathogenic nematode (EPN) EPN EPN EPN EPN EPN Target(s) Sciarids, leafminer, WFT Vine weevil Vine weevil, Vine weevil Grubs Codling moth (occasional cutworms) Hylobius sp., Lepidoptera Lepidoptera Slugs Root diseases

Main messages
Identifying suitable control agents Suitable markets for product development Enabling technologies Production and quality control End user education
(1999)

Steinernema carpocasae Steinernema carpocapsae Phasmarhabditis hermaphrodita Trichoderma harzianum Rifai (T-22)

Nemasys C Capsanem Nemaslug Trianum

EPN EPN Slug parasitic nematode Micro-organism

Specificity (IPM compatibility)

Example: viruses Mostly highly specific Mostly for Lepidoptera In vivo production, but highly potent Cases of resistance
(1998)

Fungi: example After 15 years of

and 10.2 M

what did we learn about developing new mycopesticides?

* sponsored by: CIDA, DfID, DGIS, SDC, USAID implemented by: CABI, CILSS, GTZ, IITA, IPARC www.lubilosa.org

1. The importance of working with appropriate isolates

Names change
e.g Metarhizium
Metarhizium anisopliae includes many isolates previously described as M. anisopliae var. anisopliae Previously in M. anisopliae: M. guizhouense (syn. M. taii) M. pingshaense M. acridum stat. nov. (= M. anisopliae var. acridum) M. lepidiotae stat. nov. (= M. anisopliae var. lepidiotae) M. majus stat. nov. (= M. anisopliae var. major) M. globosum sp. nov. M. robertsii sp. nov. M. brunneum
Bischoff, Rehner, Humber (2009). Mycologia 101: 512-530

Metarhizium acridum (was M. flavoviride)

Species new to science

There is more to Trichoderma than Trichoderma stromaticum T. harzianum (& T. viride) T. asperellum
Moniliophthora (Crinipellis spp) diseases of cocoa E.g. witches broom: major threat in Latin America Poor response to chemical fungicides

T. koningii

T. ovalisporum
http://nt.ars-grin.gov/taxadescriptions/keys/FrameListAllTaxa.cfm?gen=Trichoderma

2. The importance of developing appropriate technologies

3. The importance of product development: - (socio) economics and extension - registration, marketing, stewardship

Biopesticides are (or should be) specific (low impact on natural enemies and environment) Typically small (niche) markets Typically developed by small-medium sized enterprises (not agrochemical majors) Registration package - the most valuable asset?

Enabling technologies
in the public domain (P) Identification of virulent fungal isolates Mass production systems: pilot commercial Spore separation and packaging Storage techniques and models Formulations (oil-based) Application methods Field testing techniques Registration and licensing P
P X

Parallels with chemical pesticide development

Production
Packaging & storage

Formulation

P P (partial) P P X!

Strategy for use Application

Operational practicalities

similarities and contrasts

Environment, Host-pathogen interactions

Mass production options

Single stage liquid fermentation Two stage fermentation Fungus grown first in liquid medium then: Spore inoculum to solid substrate for conidiation Added to solid substrate (often grain) for conidiation Solid substrate fermentation

A commercial system
(Sylvan Industrial SSF)
Double cone blender

Fungus grown only in liquid culture

Poured into trays for conidiation

Mixing inoculum

Liquid culture & Inoculation

Production options
Capital intensive production systems Require high degree of engineering Expensive to set up Can be cost effective, but require large market/product range Labour intensive production systems Can produce high quality products Relatively cheap to set up Good for small markets Ideal for product development
Swanson D, (1997) Mem. Ent. Soc. Canada, 171, 101

Stable suspensions
for reliable ULV spraying: and subsequently emulsifiable formulations for MV / HV application

Drying, Packaging and Shelf life

Bag sealing machine

Moisture analyser

Shelf life: storage model

log10 = KE - Cwlog10m - CHt - CQt2 m moisture content (m.c.: % wb) KE (6.3) absolute longevity extrapolated from value of log10 s at 1% m.c. Cw (3.06) describes effect of m.c. on longevity CH (0.00176) and CQ (0.000703) describe the effect of temperature on longevity t temperature
Hong et al. (2001) Mycological Research 105: 597

Packaging, viability indicator

Tri-laminate foil sachets

10 years at 4oC?

If we knew then what we know now e.g. cost of application

100 g Green Muscle = 5 x 1012 conidia

Summary: biopesticides
Biopesticides products must have a clear role, work reliably, be acceptable and available; Biopesticide products and delivery systems may require a level of sophistication with parallels to chemical pesticide development Biopesticides often developed by enthusiastic scientists in research establishments and SMEs ... now more interest with pesticide restrictions enabling technologies available for development of control agents (sometimes new to science).

costs approximately $20 to produce Chemical locust insecticides approximately $10/ha Green Muscle works well at 100 g/ha if only we had done more trials at 25 g/ha ? (with high quality material)

Hindsight is a wonderful thing!