Novel Strategies for Integrating Land-Snail Pests Control of Agricultural Crops in Europe, with Projection to Latin-America and Egypt

University of Santiago de Compostela Faculty of Biology Department Zoology Santiago de Compostela La Coruña. Galicia. Spain

Novel strategies for integrating land-snail pests control of agricultural crops in Europe, with projection to Latin-America and Egypt ......

Dr José Castillejo & Dr Javier Iglesias

WORK PROGRAMME 2010 FP7 Cooperation Work Programme Food, Agriculture and Fisheries, and Biotechnology KBBE.2010.1.2-05: Integrated pest management in farming systems of major importance for Europe. Call: FP7-KBBE-2010-4 1 WORK PROGRAMME 2010

COOPERATION

THEME 2

FOOD, AGRICULTURE AND FISHERIES, AND BIOTECHNOLOGY

(European Commission C(2009) 5893 of 29 July 2009)

2 The Slug and Snail Problem….

In recent years, the problems caused by slugs, especially the grey field slug (Deroceras reirulatum), the Spanish slug (Arion lusitanicus) and the greenhouse slug (Milax gagates), have increased dramatically, as illustrated by the 70-fold increase of molluscicide usage over the last 30 years as observed in Europe. These species are a serious pest of global economic importance (South, 1992) as they have adapted well to the varied environments to which they have been introduced around the world. A. lusitanicus is polyphagous and feeds on a range of crop species as well as dumped plant material and carcasses (Wittenberg 2005). In winter wheat alone, molluscicide use, including its application, is calculated to cost some £ 20 millions annually, yet the damage to seeds and seedlings is not reliable controlled (GLEN, 1989)

3 Winter wheat, Brussels sprouts and rape crops. United Kingdom

In winter wheat, Brussels sprouts and rape crops, molluscicide use, including its application, is calculated to cost some £ 50 million annually in the United Kingdom, yet the damage to seed and seedling is not reliable controlled. 4 Sweden and Norway, Strawberry

In Sweden the species is reported from strawberry fields and grain storage facilities. No overall assessment of the economic consequences of A. lusitanicus has been Strawberry growers in Norway have reported more than made, but the species contributes to damage on several 50% loss in yield due to A. lusitanicus, but proper economic horticultural crops (Fischer and Reisschütz 1999, Speiser assessments have not been conducted yet. An example of a et al. 2001). Furthermore, there are great impediments societal effect is that home owners have been known to sell to human use of gardens as judged by the number of their property and move to slug free areas. House prices times this species make headlines in media (often under may also be affected by the presence of this. the alias “killer slug”)(Valovirta 2000). 5 Gardens

In Central Europe, Limax maximus and Arion lusitanicus are the major pest slug species, and most sales of molluscicide pellets in the home and garden market can be attributed to this species – this gives an indirect estimate of the damage they cause. Many of the European slugs and snails have been introduced to America, Australia and NZ and cause tremendous problems in their agricultural crops. 6 Arion lusitanicus

Arion lusitanicus is polyphagous and feeds on a range of crop species as well as dumped plant material and carcasses (Wittenberg 2005). In Sweden the species is reported from strawberry fields and grain storage facilities.

No overall assessment of the economic consequences of A. lusitanicus has been made, but the species contributes to damage on several horticultural crops (Fischer and Reisschütz 1999, Speiser et al. 2001). Furthermore, there are great impediments to human use of gardens as judged by the number of times this species make headlines in media (often under the alias “killer slug”)(Valovirta 2000).

7 European distribution for Arion lisitanicus, with reported agricultural damages

KBBE.2010.1.2-05: Integrated pest management in farming systems of major importance for Europe

8 European distribution for Deroceras reticulatum, with reported agricultural damages

KBBE.2010.1.2-05: Integrated pest management in farming systems of major importance for Europe

9 European distribution for Lehamania marginata, with reported agricultural damages

KBBE.2010.1.2-05: Integrated pest management in farming systems of major importance for Europe

10 Oilseed rape and cereal crops in Europe

Slugs are common pests of oilseed rape and cereal crops in Europe and are currently controlled using bait pellets that often fail to give adequate protection.

1 1 Sales of pesticides in Finland

Product index by Active Substance Metaldehyde: B&Q Slug Killer Blue Mini Pellets, Barclay Metaldehyde Dry, Barclay Tracker, Bio Slug Mini Pellets, BRITS, Doff Slugoids Slug Killer Blue Mini-Pellets, Escar-Go 6, Gastrotox Mini Slug Pellets, Gastrotox Slug Pellets, Goulding Slug Pellets, Greenfingers Slug Pellets, Hygeia Slug Pellets, Hytox Slug Pellets, Luxan Metaldehyde 5, Luxan Red 5, Metarex Green, Metarex RG, Molotov, Optimol, Pathfinder Excel, Slug Clear, Slug Killer Blue Mini-Pellets, Slug Out, Slug Pellets, Slugit Xtra, Slugtox, Stockmaster Slug & Snail Killer 12 Department for Environment, Food and Rural Affairs Nobel House. 17 Smith Square. London SW1P 3JR Telephone 020 7238 6000. Website: www.defra.gov.uk © Crown copyright 2007 13 The figure examine the detailed trends within winter wheat, which accounts for a 45% of the UK cropped area, and a significant amount of molluscicide use. 2006 report of indicators reflecting the impacts of pesticide use.

14 SPAIN

15 The New Molluscicide: Ferramol Active Ingredient: Iron phosphate: 1.0% Inert Ingredients: 99.0%

Ferramol has a low impact on the environment. Ferric phosphate, the active ingredient in Ferramol, has extremely low mammalian toxicity. It is virtually harmless to higher animals. Carabid beetles, earthworms, bees, birds and other beneficial organisms are not affected. When it biodegrades it releases iron and phosphorus into the soil which are essential plant nutrients. Although there is no aquatic toxicity from Ferramol as a matter of good practice it is better to avoid its use around water courses. Ferramol is allowed under EU regulation 2092/91 (the minimum legal standards for organic production).

16 METALDEHYDE

METAREX is a registered trademark of De Sangosse, France

At least 3 major slug species have become established and feed on vegetables (eg. lettuce, brassicas, onions, herbs, etc), arable crops and pastures.

Slugs feed vigorously above and below the ground on seeds, roots and shoots doing most of the damage at night and hiding during the day.

17 The sustainable use of pesticides

Products Containing Metaldehyde. Caution Poison

This product may be harmful to children and fatal to domestic animals if ingested. Toxic to aquatic organisms. Do not apply product near water (including garden ponds) or storm drains. Do not apply if heavy rain is expected. Toxic to birds Toxic to small wild mammals

Directive 2006/0132 (COD) of the European Parliament and of the Council of 12 July 2006 establishing a framework for Community action to achieve a sustainable use of pesticides. The specific objectives of the Thematic Strategy are:

 to minimise the hazards and risks to health and environment from the use of pesticides  to improve controls on the use and distribution of pesticides  to reduce the levels of harmful active substances including through substituting the most dangerous with safer (including non-chemical) alternatives  to encourage the use of low-input or pesticide-free crop farming, in particular by raising users' awareness, by promoting codes of good practices and consideration of the possible application of financial instruments  to establish a transparent system for reporting and monitoring the progress made towards the achievement of the objectives of the strategy, including the development of suitable indicators.

18 Pesticide use in Europe

Pesticide use in Europe. Pesticides sales in Europe are increasing. Levels of usage vary between countries. These profiles are part of an on-going series in Pesticides News that will cover all of Europe. Sources: Oppenheimer, Wolf & Donnelly, Belgium, 1997. Molluscicides sales represents 10% of all pesticides.

Directive 2006/0132 (COD) of the European Parliament and of the Council of 12 July 2006 establishing a framework for Community action to achieve a sustainable use of pesticides.

19 20 21 22 Fighting against slug and snail agricultural damages with Biocides: Molluscicides Metaldehide, Thiocarb and Ferric Phosphate

World Biocides Global biocide demand to grow 5.4% annually through 2009 World demand for biocides is projected to increase 5.4 percent per year to $6.9 billion in 2009. North America and Western Europe will remain the largest regional markets, accounting for over two thirds of demand. The Asia/ Pacific region, due mainly to continued rapid growth in China, is expected to register the fastest growth among the major regions through this decade. Eastern Europe is also expected to register above average growth, but will still account for less than five percent of global demand. In more mature markets, such as Japan, the United States and Western Europe, advances will be modest, with gains spurred by the replacement of traditional products with higher value formulations offering a combination of broad-spectrum efficacy, low toxicity, minimal effect on finished product quality and reduced environmental impact. Much of this shift will be prompted by the sizable regulatory framework under which the biocide industry operates. Many biocides are synthetic, but a class of natural biocides, derived from e.g. bacteria and plants

23 24 http://ec.europa.eu/food/plant/protection/evaluation/exist_subs_rep_en.h tm

25 http://ec.europa.eu/food/plant/protection/evaluation/exist_subs_rep_en.h tm

26 27 PART A: EXISTING ACTIVE SUBSTANCES DIVIDED INTO FOUR LISTS FOR PHASED EVALUATIONS

28 The current strategy: kill the animal

Lonza Metaldehyde Ferramol Bayer Iron phosphate Methiocarb

29 BAYER STRATEGY It’s correct the Bayer, Lonza and Ferramol strategy?

But, this strategy ignore the laid land snail’s egg on the soil: one egg, one slug. 1 egg = 1 slug

What about the land snail eggs?

31 The novel strategies for integrate land snail pests control of agricultural crops

Concept and objectives

To introduce in a series of crops, new strategies for the integrated land snail pest control.

These strategies are based in the deep knowledge of the pest´s biology and ecology, so that the abundance and the activity periods can be predicted to elaborate an integrated pest control system and take decisions that can be applied in every develop phase (juvenile, adult, senile or eggs).

Thanks to this the farmer will know: . when to apply the traditional molluscicides (to destroy the land snails) . when to use the molluscicides ovicidal (to destroy the egg lays) .when to apply the biological control through parasite nematodes or when to use the trap-plants

32 First to know the land snail biological cycle to find the weak point where to apply our strategy

Our strategies are based on:

 To understand the land snails biological cycle in the crops study areas.

 To understand the land snails activity in function of the climatic variables and the crop type.

 To destroy the land snail´s egg-lays thanks to the plant extracts and non residual standard agrochemicals collateral effect.

 To rationalize standard molluscicides consumption in standard farming.

 To introduce cattle and swine slurry and trap-plants as control methods in organic farming.

 To understand the collateral effects of the products used in this integrated pest control methods.

33 Controlling Slugs and Snails

AGROCHEMICALS Laboratory and field experiments with low- chemical against slugs NEMATODES Search for warm-adapted Trap-plants to dissuade strain of slug- Slug and Snails parasitic

34 Controlling Snail and Slug’ Eggs

AGROCHEMICALS Laboratory and field test with Chemical compound with horticultural application

MANURE PLANT EXTRACTS Laboratory and Laboratory test with Digitalis purpurea L., Eucalyptus globulus field test with Labill., Euphorbia helioscopia L., Foeniculum vulgare Miller., Laurus nobilis L., Rosmarinus Suidae and officinalis L., Rubia peregrina L. and Ruta Bovidae: graveolens L Different concentrations and origins

Deroceras reticulatum eggs 35 Collateral effects on soil fauna Soil Infaune monitored

OBJETIVES: . Assess the impact of some bio-molluscicides on non- Round Worms target soil invertebrates

BIO-MOLLUSCICIDES TESTED: • Cow slurry/Slug Eggs Killer • Herbicides/Slug Eggs Killer • Plant extracts Slugs

Snails

Earthworms

Coleoptera Springtails WP.1. Land Snail Biological Cycle Based on: Crop Phenology and Meteorological conditions

Biological Cycle Crops Phenology Cultivation, tilling, ploughing

Variables Assembling Statistical model to predict activity Meteorology

Activity Maps 37 STATISTICAL MODEL TO PREDICT ACTIVITY NATURAL

Biological Studies on Land Snails and crops

20 15 10 5 0 C Y C L E

38 ACTIVITY TRAP PLANTS STRATEGY

Sow thistle Sonchus oleraceus L. Implications of the new strategy

Winter Spring Summer Autumn E F M A M J J A S O N D CONSEQUENCES OF THE STATISTICAL MODEL RIGHT MODELLING MOMENT

PREDICTION OF HIGHER HIGHER RISK EFFICACY PERIODS ECONOMICAL ECOLOGICAL ADVANTAGES ADVANTAGES FARMER APPLY CONTROL METHODS BEFORE SLUG ATTACK FEWER PRODUCT FEWER APPLICATIONS

FEWER TOXIC RESIDUES ON FOOD USEFULL INVERTEBRATES FEWER PRODUCT FEWER PLANT DAMAGE KILLED = SAVE MONEY TOXIC COMPOUNDS IN =EARN MONEY WATER

41 Our strategy versus Bayer, Lonza, Ferramol stratgy

Lonza Metaldehyde Ferramol Iron phosphate Bayer Methiocarb

42 Malaterra Laboratory at USC Terrestrial Applied Malacology Lab 43 44 Room for producing slug eggs. Controlled environment

45 Outside mini plots with anti escape system for testing bio-molluscicides

46 47 Video Camera with Infrared light

Weather Station 48 Video Tracking System

WP. 1 .-Biological Cycle

Video Camera

53 54 Weather Station

55 Weather Station

Digital Video Recorder

56 Weather Station

57 Work Package 1. Land Snail Biological Cycle. Size, structure and dynamic.

58 WP. 2. Slugs diet. Trap Plants

Captura de 20 babosas/mes entre diciembre de 1999 y noviembre de 2001 Tratamiento de los individuos

ESÓFAGO 4 1 Extracción To weigh del buche

Crop. Stomac

5 2 Obtención Sacrificio del contenido a 50ºC estomacal 6 Masa del CONDUCTOS DE LA contenido GLÁNDULA DIGESTIVA 7 3 Almacenamiento Disección en 2 ml HCl 1N Determinación de la dieta

Se identificaron Se tomo una muestra Se analizaron al y midieron cada uno de de 0,24 ml de cada microscopio los fragmentos contenido estomacal óptico Determinación de la dieta

Célula epitelial

25 μ 50 μ

Stellaria media Ranunculus repens Tricoma Estoma

Célula epitelial 25 μ 50 μ

Mentha suaveolens Stachys arvensis

Para ello se usaron las células epiteliales, estomas y tricomas Determinación de la dieta

Una vez identificados los fragmentos, estos fueron medidos. Determinación de la dieta Determinación de la dieta

0,08 mm2

Área superficial (mm2) de cada tipo de alimento x100 la contribución (%) de cada

tipo de alimento a la dieta = Área superficial (mm2) de todos los fragmentos de la muestra University of Santiago de Compostela Faculty of Biology Spain

Agrochemical Compounds as bio-molluscicide for Controlling Slug damages

Dr José CASTILLEJO & Dr Javier IGLESIAS

* Investigación financiada por: • la Unión Europea. Nº de Proyecto: FAIR5-PL97-3355. Título: “Novel technologies for integrated control of slug damage in key horticultural crops” • la Xunta de Galicia. Nº de Proyecto: PGIDT00AGR20001PR Título: “Alternativas a los molusquicidas comerciales actuales” Egg Selection

Selection process

Stereo Microscopy Effect of Chemical Compound

Alive

Product

Selection

Dead Dead Agrochemical Compounds Filter Paper

Standard Soil Other material Filter paper 1 ml of product(mg/cm2)

Dead 1 Alive 0 Hatch  Dry off

1 ml of Destilled water

D. reticulatum eggs

+ Activity / tested in standard soil

– Activity / no tested in standard soil Certrol H CHARACTERISTICS: Comercial name Certrol H Composition Ioxynil 12% ; MECOPROP 36% . Manufacture Comercial Quimica MASSO S.A Activity Herbicide

10 7,5 1 0,75

(logscale)

2 0,1 0,075

0,01 0,0075

0,001 Dose mg/cmDose 0 5 10 15 20 25 Time in days Filter paper Standard-soil

Fig 1 Time and dose relatioship for wich 100% mortality was obtained. Tordon 101

CHARACTERISTICS: Comercial name TORDON 101 Composition Plicoran 6% and Tri-isopropanolamina 24% Manufacture RHONE-POULENC AGRO, S.A Activity Especific herbicide, used in temperature >25ºC.

10 4,7

1 0,47 0,47

(log scale) 2 0,1 0,047 0,047 0,01 0,0047

Dose mg/cmDose 0,001 0 5 10 15 20 25 Time in days

Filter paper Standard-soil

Fig 2. Time and dose relatioship for wich 100% mortality was obtained. Lugsamag-N

CHARACTERISTICS: Comercial name Lugsamag-N Composition 6,2% total nitrogen nitric & 8,8% of MgO oligoelements 0.025 B ; 0.01 Cu ; 0.1 Fe ; 0.07 Mn ; 0.003 Mo ; 0.015 Zn Manufacture UQSA S.A Activity soil nutrent, used to reduce carency of microelements.

10 3 3 1

(log scale) 2 0,3 0,1 0,03

Dose mg/cmDose 0,01 0 5 10 15 20 25 Time in days Filter paper Standard-soil

Fig 3. Time and dose relatioship for wich 100% mortality was obtained. Garlon Gs CHARACTERISTICS: Comercial name Garlon GS Composition 9% Triclopir and 3% Fluroxipir Manufacture DOWELANCO Iberica S.A. Activity Herbicide used in horticultural crops against bad grass.

10 1,8

1 0.18

(log scale) 2 0,1 0,18 0,018 0,01 0,0018

Dose mg/cmDose 0,001 0 5 10 15 20 25 Time in days

Filter paper Standard-soil

Fig. 4 Time and dose relatioship for wich 100% mortality was obtained. Chas-48

CHARACTERISTICS Comercial name Chas 4 Composition Clorpirifos 48% & aroamatic hidrocarbures. Manufacture AGRODAN S.A Activity Insecticide

10 7

1 0,7 0,7

(log scale)

2 0,1 0,07

0,01

0,001

Dose mg/cmDose 0 5 10 15 20 25 Time in days Filter paper Standard-soil

Fig. 5 Time and dose relatioship for wich 100% mortality was obtained. 100

1,57 1 1,178

(log scale) (log 0,75 0,55

2 0,23 0,157 0,1 0,047 0,055

0,01 0,023

Dosemg/cm

0,0023 0,001 0 5 10 15 20 25 Time in days

METALDEHIDE AMIGO DIMILIN AZADIRACHTIN NOMOLT SAPONIN BACTOSAN

Fig.11 Time and dose relatioship for wich 100% mortality was obtained. Results on slug eggs: slurry

300

275

250

-2 225 pig-slurry 200 caw-slurry

in mg cm mg in 50 175

150

125

100 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Exposure in days University of Santiago de Compostela Faculty of Biology Spain

SCREENING ON PLANT EXTRACTS FOR OVICIDAL ACTIVITY AGAINST Deroceras reticulatum EGGS

José CASTILLEJO & Javier IGLESIAS

AGAINST EGGS

ACTIVE PRODUCT AGAINST ADULTS

PLANT EXTRACTS

PLANTS

SYNTHETIC HOW and PRODUCT WHEN to ® apply it ? COMMERCIALIZATION PATENT 79 Preparing the extracts for testing:

. Plants were collected and identified . Different parts of the plants were separated:

Rhizomes Leaves Fruits Testing the plants:

. 5 doses of the extract and one control. The doses were arranged in a ten-fold geometric series . 3 replicates per dose . 3 to 5 eggs per replicate . 1 ml of extract was applied to each dish . After dry were remoistened with 1 ml of water Euphorbia helioscopia L.

. Common name: Sun spurge (Euforbia) . Constituents: euphorbone; 12- deoxyphorbol; wax; resin; lignin; basorrin; volatile oils; euphorbo- resene; euphorbic acid; SPURGE: Any of various euphorbiaceous plants of the calcium malate genus Euphorbia that have milky sap and small flowers typically surrounded by conspicuous bracts. Some species have purgative properties Euphorbia’s extracts tested

LEAVES STEAMS ROOTS FRUIT

FRESH DRY FRESH DRY FRESH DRY FRESH DRY

Fp Fp Fp Fp Fp Fp Fp Fp

100,000         W A 80,000        

T 50,000         E 40,000         R (ppm) 8,000        

ACENTO NE / 100,000 WATER         (ppm) Ruta graveolens L.

. Common name: Rue (Ruda)

. Constituents: furocouamarines, rutin, volatile oils, alkaloids, coumarines.

RUE: any rutaceous plant of the genus Ruta, esp. R. graveolens, an aromatic Eurasian shrub with small yellow flowers and evergreen leaves which yield an acrid volatile oil, formerly used medicinally as a narcotic and stimulant. Archaic name: herb of grace Ruta’s extracts tested

Fp = Filter paper LEAVES STEAMS FLOWERS Ss = Standard soil FRESH DRY FRESH DRY FRESH DRY Fp Ss Fp Ss Fp Ss Fp Ss Fp Ss Fp Ss 100,000    -    -    - W 80,000             A 50,000    -    -    - T 40,000             10,000 E   - -    -   - - 8,000             R Ruta’s dry leaves (ppm) 1,000   - -   acetone /- water(7:3) extracts - - 100   - -    Mortality- over time - - A 100,000        -     C 100 80,000 E      80       T 60 50,000 O    -  40  -     N 40,000 % mortality 20 E             0 / 10,000    -    -    - W 0 5 10 15 20 25 A 8,000         Time in days    T 1,000 E   - -   - 80000 ppm- 40000 ppm  20000 -ppm - 10000 ppm 5000 ppm R 100   - -   - -   - - (ppm) DEFINITIVE Ruta’S EXPERIMENTS ON FILTER PAPER

LEAVES DRY Fp W 80,000  A 40,000  T 20,000  E R 10,000  (ppm) 5,000  80,000  ACETONE / 40,000  WATER 20,000  (ppm) 10,000  5,000  Ruta’s dry leaves Water extracts Mortality over time

100

% mortality 20

0 0 5 10 15 20 Time in days

80000 ppm 40000 ppm 20000 ppm Dose-response curve/ LD50

100

60 22939 32707 40 18455

% mortality 20

0 1000 10000 100000 Dose in ppm (log scale)

5 days 7 days 10 days LD50 Ruta’s dry leaves acetone / water (7:3) extracts Mortality over time

100 80 60 40

% mortality 20 0 0 5 10 15 20 25 Time in days

80000 ppm 40000 ppm 20000 ppm 10000 ppm 5000 ppm Dose-response curve

100

60 7862 27841 16907 40

% mortality 20

0 1000 10000 100000 Dose in ppm (log scale)

2 days 3 days 5 days LD50 Digitalis purpurea L.

. Common name: Foxglove (España: dedalera) . Constituents:

1. glucosides : digitoxin, digitalin, digitalein, digiton

2. volatile oil

3. fatty matter

4. gum

5. sugar

FOXGLOVE: any Eurasian scrophulariaceous plant of the genus Digitalis, esp. D. purpurea, having spikes of purple or white thimble-like flowers. The soft wrinkled leaves are a source of digitalis Digitalis’ extract tested

Fp = Filter paper LEAVES Ss = Standard soil FRESH Fp

80,000  WATER 40,000  (ppm) 8,000  ACETONE / WATER 100,000  (ppm)

Searching for Nematode Strains The Spanish Phasmarhabdities strain

The nematodes found on Iberian Peninsula slugs was identified as Phasmarhabditis sp. They appear under four forms: 1. Dauer larva, with differents degrees of develoopment. 2. Small females (<100 microm. long) full of eggs 3. Big size males (twice than female). 4.Big size Females measurent between 1800-2500 microms. Clearly different from the another one. The specific separation between P. hermaphrodita and P. neopapillosa is not clear, the Spanish morpho could be new specie. Our strategy versus Bayer, Lonza, Ferramol stratgy

Lonza Metaldehyde Ferramol Iron phosphate Bayer Methiocarb

97 PARCEL miniPLOTS

2.6 m

2 m2 1.4 m

• Six replicates for each crops/test • Six controls plots/random selection • Monthly sampling 1.4 m AREA FOR INFAUNAE SAMPLING

Centre Phasmarhabditis Metaldehyde Ioxynil (Herbicide) Phasmarhabditis + Ioxynil Periphery Phasmarhabditis + Metaldehyde (Nothing)

2 m2

14 m2

Before starting experiments the soil fauna was assessed on every plot and its spatial distribution was homogeneous 100 Our Strategy implications: when to apply the Molluscicide?: in Summer

Winter Spring Summer Autumn E F M A M J J A S O N D

Eggs Eggs Eggs Eggs

This strategy helped to reduce pesticide use on crops applying the necessary quantity at the right time, not introducing new chemicals in agricultural crops, but taking advantage of the farmers’ standard used non residual agrochemicals just giving it a different use or using the favourable side-effects. Thereby decreasing the amount of toxic agents that may be harmful to humans and to wildlife and soil. 101 THE NEW PROJECT

WHAT TO DO..... If the project match inside the next Brussels call: 1. Define the number of appropriate WPs 2. Rewrite the WP for specialists 3. Coordination of each WP 4. Looking for European teams with capacity to carry out all WPs 5. The number of participants. Countries 6. What about Egypt, Tunis as participants 7. What about Hispano America as participants

102 • Defining and rewriting the Work Packages

103 104 Marivonne PERT WP1 Land snails ‘ biological

cycle

plants..

trap WP3 Statistical models to predict activity

WP2

WP6 andWP6 WP7

WP Exploitation WP

Field experiments Field experiments

composition,

WP4 WP5 Bioovicide Diet Biomoll tests Agrochem tests

WP Collaboration and Dissemination

WP Project Management

Project Project Start End Dr Albert Ester PAV. Edelhertweg P.O. Box 430 8200 AK Lelystad. The Netherlands

Dr. Solveig Haukeland. Bioforsk Norwegian Institute for Agricultural and Environmental Research Fr. A. Dahlsvei 20, N-1432 Ås Norway (Noruega) Dr. Olaf Schmidt Ph.D. (Senior Lecturer) UCD School of Agriculture, Food Science and Veterinary Medicine. University College Dublin. Belfield, Dublin 4. Ireland

Dr Georges Dussart Dr. Grita Skujienė Canterbury Christ Church University Department of Zoology North Holmes Road Faculty of Natural Sciences Canterbury Vilnius University Kent, CT1 1QU Ciurlionio 21/27, United Kingdom 03101 Vilnius Lithuania Dr. André Chabert ACTA. 4 place Gensoul 69287 Lyon Cedex 12 France

Dr Maryvonne Charrier, MC Dr Eva Knop Université de Rennes 1- UMR EcoBio 6553 University of Bern Campus de Beaulieu, Bât. 14A Institute of Ecology and Evolution F-35042 RENNES Cedex Baltzerstrasse 6 France CH-3012 Bern. Switzerland

Dr. José Castillejo & Dr. Javier Iglesias Departamento de Zoología Facultad de Biología Universidad de Santiago de Compostela 15782 Santiago de Compostela Syngenta Crop Protection La Coruña. Galicia. España Syngenta Crop Protection UK Ltd. CPC4 Capital Park. Fulbourn Dra. Mª Mercedes Ortega Hidalgo Cambridge CB21 5XE. UK Dpto. Gent. Antrop. Física y Fisiología Animal E-mail: [email protected] Facultad de Ciencia y Tecnología Universidad del Pais Vasco. España

106 Dr. Intidhar ABBES Biodiversité et Biologie des Populations Sciences Biologiques Faculté des sciences de Tunis Université Tunis El Manar, 2092, Tunisie

Dr Maha Shoieb uez canal university College of Agriculture Plant Protection Department Ismailia, Egypt

107 Dra. Lenita de Freitas Tallarico Lenita de Freitas Tallarico (Biologist) Laboratorio de Dr. Galileo Rivas Parasitología/Malacología Líder Programa de Producción Agroecológica de Cultivos Instituto Butantan Alimenticios Avda. Voital Brasil, 1500 Centro Agronómico Tropical de Investigación y Enseñanza, CATIE CEP-05503-900 Sao Paulo. Brasil División de Investigación y Desarrollo 7170 Turrialba, Cartago. Costa Rica

Dra. Luz Elena Velásquez Trujillo Programa de Estudio y Control de Enfermedades Tropicales –PECET Universidad de Antioquia Medellín, Colombia

Dr. BARRAGAN YANEZ ALVARO RODRIGO Pontificia Universidad Católica del Ecuador Facultad de Ciencias Exactas y Naturales ESCUELA DE CIENCIAS BIOLÓGICAS 12 de Octubre entre Patria y Veintimilla Quito. Ecuador

Dr. Enrique La Hoz Brito. Director General de Investigación. DIA - INIAAv. La Molina (Ex Av. La Universidad) No. 1981 Lima 1 Perú

Dr. Sergio Letelier V. Laboratorio de malacología Museo Nacional de Historia Natural Santiago de Chile Interior Quinta Normal s/n, Casilla 787 56-02-6804648 Santiago de Chile Chile Dra. Carla Salvio Faculty of Agricultural Sciences National University of Mar del Plata Experimental Station of National Institute of Agricultural Technology (INTA) C.C: 276 (7620) Balcarce. Argentina 108 Syngenta Crop Protection Syngenta Crop Protection UK Ltd. CPC4 Capital Park. Fulbourn Cambridge CB21 5XE. UK E-mail: [email protected] 109 PROJECT MANAGEMENT STRUCTURE

Beware!. Coordination for each WP WP 2 Manager: Participant ? Participants 1, 2 ,3, 4, 5, 6, 7, WP 1 8 WP 3 Manager: Participant ? Manager: Participant ? Participants 1, 2 ,3, 4, 5, 6, 7, Participants 1, 2 ,3, 4, 5, 6, 7, 8 8

COORDINATION Participant 1 WP ???? WP 4 DELIVEREBLES Manager: Participant ? Manager: Participant ? •Statistical model to predic activity Participants 1, 2 ,3, 4, 5, 6, 7, Participants 1, 2 ,3, 4, 5, 6, 7, • Molluscicides ovicicides 8 8 • Nematode zooparasitic • Integrate package for organic crops • Integrate package for convencional crops

WP 7 WP 5 Manager: Participant ? Manager: Participant ? Participants 1, 2 ,3, 4, 5, 6, 7. Participants 1,2,3,4, 5, 6, 7, 8 8 WP 6 Manager: Participant ? Participants 1, 2 ,3, 4, 5, 6, 7, 8 WP 2 RELATIONSHIP TASKS and Participants DIET COMPOSITION TRAP PLANTS

Participant 1, 2 ,3, 4, 5, 6, 7, 8 WP 1 WP 3 LAND SNAIL’S BIOLOGICAL STATISTICAL ACTIVITY CYCLE MODEL

Participant 1, 2 ,3, 4, 5, 6, 7, Participant 1, 2 ,3, 4, 5, 6, 7, 8 8

WP 4 • WP 9 Statistical model to predic activity BIOMOLLUSCICIDES TESTS NEMATODO ZOOPARASITIC • Molluscicides ovicicides COLLATERAL EFFECTS Phasmarhadities • Nematode zooparasitic • Integrate package for organic crops Participant 1, 2 ,3, 4, 5, 6, 7, Participant: Latino America • Integrate package for convencional crops 8

WP 7 & 8 WP 5 FIED EXPERIMENTS BIO OVICIDAS CONVENTIONALLY, AGROCHEMICALS TESTS ORGANICS CROPS WP 6 Participant 1, 2 ,3, 4, 5, 6, 7. Participant 4, 5, 6, 7, 8 PIG AND COW MANURE 8 AS BIO OVICIDES TESTS

Participant 1, 2 ,3, 4, 5, 6, 7, 8 WP 2 RELATIONSHIP TASKS and Participants DIET COMPOSITION TRAP PLANTS

Participant 1, 2 ,3, 4, 5, 6, 7, 8 WP 1 WP 3 LAND SNAIL’S BIOLOGICAL STATISTICAL ACTIVITY CYCLE MODEL

Participant 1, 2 ,3, 4, 5, 6, 7, Participant 1, 2 ,3, 4, 5, 6, 7, 8 8

WP 4 WP 9 BIOMOLLUSCICIDES TESTS NEMATODO ZOOPARASITIC intercommunication COLLATERAL EFFECTS Phasmarhadities Participant 1, 2 ,3, 4, 5, 6, 7, Participant: Latino America 8

Participant 1, 2 ,3, 4, 5, 6, 7, 8 RELATIONSHIP Participants and TASKS Participant 3 Research Centre Country WP. 1, 2 ,3, 4, 5, 6, 7

Participant 2 Participant 4 Research Centre Research Centre Country Country WP. 1, 2 ,3, 4, 5, 6,7 WP. 1, 2 ,3, 4, 5, 6

Participant 1 Participant 8 Participant 4 Coordinator Research Centre Research Centre Country Country USC WP. 1, 2 ,3, 4, 5, 6,7 WP. 1, 2 ,3, 4, 5, 6, 7 Spain WP. 1, 2 ,3, 4, 5, 6, 7, 8

Participant 7 Participant 5 Research Centre Research Centre Country Country WP. 1, 2 ,3, 4, 5, 6, 7 WP. 1, 2 ,3, 4, 5, 6,7 Participant 6 Research Centre Country WP. 1, 2 ,3, 4, 5, 6, 7 RELATIONSHIP Participants and TASKS Participant 1 Research Centre Participant 2 Country Research Centre Participant 10 WP. 1, 2 ,3, 4, 5, 6, 7 Country Research Centre WP. 1, 2 ,3, 4, 5, 6 Country WP. 1, 2 ,3, 4, 5, 6,7

Participant 3 Research Centre Participant 9 Country Research Centre WP. 1, 2 ,3, 4, 5, 6,7 Country WP. 1, 2 ,3, 4, 5, 6, 7 intercommunication Participant 4 Research Centre Participant 8 Country Research Centre WP. 1, 2 ,3, 4, 5, 6,7 Country WP. 1, 2 ,3, 4, 5, 6

Participant 7 Participant 5 Research Centre Research Centre Country Country WP. 1, 2 ,3, 4, 5, 6, 7 WP. 1, 2 ,3, 4, 5, 6 Participant 6 Research Centre Country WP. 1, 2 ,3, 4, 5, 6, 7 Participant 1 Research Centre Participant 2 Country Research Centre Participant 10 WP. 1, 2 ,3, 4, 5, 6, 7 Country Research Centre WP. 1, 2 ,3, 4, 5, 6 Country WP. 1, 2 ,3, 4, 5, 6,7

Participant 3 Research Centre Participant 9 Country Research Centre WP. 1, 2 ,3, 4, 5, 6,7 Country WP. 1, 2 ,3, 4, 5, 6, 7 Syngente Provide non residual agrochemicals for testing Participant 4 Research Centre Participant 8 Country Research Centre WP. 1, 2 ,3, 4, 5, 6,7 Country WP. 1, 2 ,3, 4, 5, 6

Participant 5 Participant 7 Research Centre Research Centre Country Country Participant 6 WP. 1, 2 ,3, 4, 5, 6, 7 WP. 1, 2 ,3, 4, 5, 6 Research Centre Country WP. 1, 2 ,3, 4, 5, 6, 7 Guidance for the time we should expect for each work package SIMULATION: Gantt Chart showing the timing of the different WPs . 1 person-month = 135 productive hours per month Work Task Number of Number of Total Hours Person- Packge Nº Work Packages Title Number of Hours per day days per person per WP or month Nº month month participating Task Hours/135 Land Snail Biological Cycle. WP.1 the size, structure and dynamics of their 8 5 24 2 1920 14.2 populations Land Snails diet composition. Trap plants WP.2. 12 5 24 2 2880 21.3

Statistical Model to predict land snails activity WP.3. 12 5 24 2 2880 21.3

Bio pesticides, Bio Molluscicides, Bio Ovicides Plants Extracts. Laboratory test on paper filter Task 4.1. Standard soil 4 10 30 2 2400 17.7

Mini plots tests on horticultural soil Task 4.2. 2 10 24 2 960 7.1 WP.4.

Collateral effects on soil fauna Task 4.3. 2 10 18 2 720 5.3 Chemicals analysis to search the active principle of plant extracts with mulliscicide and ovicide Task 4.4. 2 10 18 2 720 5.3 activity

Laboratory test to find non residual Task 5.1. agrochemicals with ovicidal activity 4 10 25 2 1920 14.2

Field experiments on conventionally horticultural crops to evaluate the efficacy of Task 5.2. 4 10 18 2 1440 10.6 WP.5. selected agrochemical as molluscicide-ovicide Field trials to evaluate the collateral effects on soil Fauna and border effect on wild land snails Task 5.3. of ovicide agrochemicals. 1 10 18 2 360 2.6

For carrying out the project. People involved: 116 • 1 person full time dedication. PhD student + 2 person partial time dedication. Staff University Manure Laboratory test on land snail eggs Task 6.1. 2 10 18 2 720 5.3 for accurate Ovicidal concentration

Field experiments to evaluate the efficacy Task 6.2. of cow and pig manure as slug eggs 2 10 24 2 960 7.1 control for key organic horticultural crops

WP.6. Field analysis to investigate the collateral Task 6.3. effect on soil fauna and border effect on 1 10 18 2 360 2.6 wild land snails of cow and pig manure

Field experiments to use tramp-plants as deterrent method to protect organic Task 6.4 1 5 24 2 240 1,7 horticultural crops alone and in combination of cow and pig manure

Field experiments in conventionally crops to evaluate the efficacy of ovicide WP. 7 10 24 2 1920 14.2 agrochemicals alone and in combination 4 with other commercial molluscicides

Field experiments in organics horticultural crops to evaluate the efficacy of organic WP. 8 4 10 24 2 1920 14.2 molluscicides-ovicides and the use of plant-traps To identify improved strains of Phasmarhabditis nematodes which are WP. 9 1 10 36 2 720 5.3 more effective biocontrol agents of larger slug species in Hispano-America.

TOTAL person-months 209.7 117 For carrying out the project. People involved: • 1 person full time dedication. PhD student • 2 person partial time dedication. Staff University 118 Gantt Chart showing the timing of the different WPs and their components.

Work Task Lead Lead Total Packge Nº Work Packages Title Type activity participant Participant Person- Start month End month Nº nº short name months Land Snail Biological Cycle. WP.1 the size, structure and dynamics of their RTD 1 24 populations Land Snails diet composition. Trap plants WP.2. RTD 1 24

Statistical Model to predict land snails WP.3. activity RTD 13 36

Bio pesticides, Bio Molluscicides, Bio Ovicides Task 4.1. Plants Extracts. Laboratory test on paper RTD 1 30 filter Standard soil Mini plots tests on horticultural soil Task 4.2. RTD 13 36 WP.4. Collateral effects on soil fauna Task 4.3. RTD 19 36

Chemicals analysis to search the active principle of Task 4.4. RTD 19 36 plant extracts with mulliscicide and ovicide activity Laboratory test to find non residual Task 5.1. agrochemicals RTD 1 24 with ovicidal activity Field experiments on conventionally horticultural crops to evaluate the efficacy of Task 5.2. RTD 13 30 selected agrochemical as WP.5. molluscicide-ovicide

Field trials to evaluate the collateral effects on soil Task 5.3. Fauna and border effect on wild land snails RTD 19 36 of ovicide agrochemicals. 119 120