pests control of agricultural crops in

(1)

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

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

Dr José Castillejo & Dr Javier Iglesias

(2)

2

WORK PROGRAMME 2010

COOPERATION THEME 2

FOOD, AGRICULTURE AND FISHERIES, AND BIOTECHNOLOGY (European Commission C(2009) 5893 of 29 July 2009)

(3)

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)

The Slug and Snail Problem….

3

(4)

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.

Winter wheat, Brussels sprouts and rape crops. United Kingdom

4

(5)

Strawberry growers in Norway have reported more than 50% loss in yield due to A. lusitanicus, but proper economic assessments have not been conducted yet. An example of a societal effect is that home owners have been known to sell their property and move to slug free areas. House prices may also be affected by the presence of this.

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).

Sweden and Norway, Strawberry

5

(6)

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.

Gardens

6

(7)

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).

Arion lusitanicus

7

(8)

European distribution for Arion lisitanicus, with reported agricultural damages

8 KBBE.2010.1.2-05: Integrated pest

management in farming systems of major importance for Europe

(9)

European distribution for Deroceras reticulatum, with reported agricultural damages

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

(10)

European distribution for Lehamania marginata, with reported agricultural damages

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

(11)

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.

Oilseed rape and cereal crops in Europe

1 1

(12)

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

(13)

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

(14)

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

(15)

15

SPAIN

(16)

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

(17)

METALDEHYDE

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.

METAREX is a registered trademark of De Sangosse, France

17

(18)

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.

The sustainable use of pesticides

18

(19)

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)

20

(21)

21

(22)

22

(23)

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

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

23

(24)

24

(25)

25

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

tm

(26)

26

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

tm

(27)

27

(28)

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

(29)

29 The current strategy: kill the animal

Bayer Methiocarb

Lonza

Metaldehyde Ferramol

Iron phosphate

(30)

BAYER STRATEGY

(31)

31 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?

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

(32)

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

(33)

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.

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

33

(34)

NEMATODES Search for warm-adapted

strain of slug- parasitic

AGROCHEMICALS Laboratory and field experiments with low-

chemical against slugs

Controlling Slugs and Snails

Trap-plants to dissuade Slug and Snails

34

(35)

Controlling Snail and Slug’ Eggs

MANURE Laboratory and

field test with Suidae and

Bovidae:

Different concentrations and

origins

AGROCHEMICALS

Laboratory and field test with Chemical compound with horticultural

application

PLANT EXTRACTS Laboratory test with

Digitalis purpurea L., Eucalyptus globulus Labill., Euphorbia helioscopia L., Foeniculum vulgare Miller., Laurus nobilis L., Rosmarinus officinalis L., Rubia peregrina L. and Ruta graveolens L

Deroceras reticulatum eggs 35

(36)

OBJETIVES:

 Assess the impact of some bio-molluscicides on non- target soil invertebrates

Soil Infaune monitored

BIO-MOLLUSCICIDES TESTED:

• Cow slurry/Slug Eggs Killer

• Herbicides/Slug Eggs Killer

• Plant extracts

Round Worms

Slugs

Snails

Earthworms

Springtails Coleoptera

Collateral effects on soil fauna

(37)

37

WP.1. Land Snail Biological Cycle

Based on: Crop Phenology and Meteorological conditions

Biological Cycle Cultivation, tilling, ploughing

Variables Assembling Statistical model to predict activity Meteorology

Crops Phenology

Activity Maps

(38)

C Y C L E

ACTIVITY

Biological Studies on Land Snails and crops

NATURAL

STATISTICAL MODEL TO PREDICT ACTIVITY

0 5 10 15 20

38

(39)

Sow thistle Sonchus oleraceus L.

TRAP PLANTS STRATEGY

(40)

Implications of the new strategy

Winter Spring Summer Autumn

E F M A M J J A S O N D

(41)

MODELLING

RIGHT MOMENT

PREDICTION OF HIGHER RISK

PERIODS

HIGHER EFFICACY

ECOLOGICAL ADVANTAGES ECONOMICAL

ADVANTAGES

FEWER PRODUCT

= SAVE MONEY

FEWER PLANT DAMAGE

=EARN MONEY

FEWER PRODUCT FEWER APPLICATIONS

FEWER

TOXIC RESIDUES ON FOOD USEFULL INVERTEBRATES

KILLED

TOXIC COMPOUNDS IN WATER

FARMER APPLY CONTROL METHODS BEFORE SLUG

ATTACK

CONSEQUENCES OF THE STATISTICAL MODEL

41

(42)

42

Our strategy versus Bayer, Lonza, Ferramol stratgy

Bayer Methiocarb

Lonza

Metaldehyde Ferramol

Iron phosphate

(43)

43

Malaterra Laboratory at USC

Terrestrial Applied Malacology Lab

(44)

44

(45)

45

Room for producing slug eggs. Controlled environment

(46)

46

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

(47)

47

(48)

48 Video Camera with Infrared light

Weather Station

(49)

Video Tracking System

(50)

(51)

(52)

(53)

53 Video Camera

WP. 1 .-Biological Cycle

(54)

54

(55)

55 Weather Station

(56)

56 Weather Station

Digital Video Recorder

(57)

57 Weather Station

(58)

58

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

(59)

WP. 2. Slugs diet. Trap Plants

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

(60)

Tratamiento de los individuos

Crop. Stomac ESÓFAGO

CONDUCTOS DE LA GLÁNDULA DIGESTIVA

1 To weigh

2 Sacrificio

a 50ºC

3 Disección

5 Obtención del contenido

estomacal 6 Masa del contenido

7 Almacenamiento en 2 ml HCl 1N

4 Extracción

del buche

(61)

Determinación de la dieta

Se tomo una muestra de 0,24 ml de cada contenido estomacal

Se analizaron al microscopio

óptico

Se identificaron y midieron cada uno de

los fragmentos

(62)

Determinación de la dieta

25 μ

Mentha suaveolens Stellaria media Célula

epitelial

Estoma

Célula epitelial

50 μ

Tricoma Ranunculus repens

Stachys arvensis

Para ello se usaron las células epiteliales, estomas y tricomas

(63)

Determinación de la dieta

Una vez identificados los fragmentos, estos fueron medidos.

(64)

Determinación de la dieta

(65)

Determinación de la dieta

Área superficial (mm

²

)

de cada tipo de alimento x100

Área superficial (mm

²

) de todos los fragmentos de la muestra

la contribución (%) de cada tipo de alimento a la dieta

0,08 mm

²

=

(66)

Agrochemical Compounds as bio-molluscicide for Controlling Slug damages

Dr José CASTILLEJO & Dr Javier IGLESIAS

University of Santiago de Compostela Faculty of Biology

Spain

*

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”

(67)

Egg Selection

Stereo Microscopy

Selection process

(68)

Effect of Chemical Compound

Product

Alive

Dead Dead

Selection

(69)

Standard Soil

Agrochemical Compounds Filter Paper

Other material

(70)

Filter paper

1 ml of product(mg/cm ² )

Dry off

1 ml of Destilled water

D. reticulatum eggs

Dead 1 Alive 0 Hatch 

+ Activity / tested in standard soil

– Activity / no tested in standard soil

(71)

CHARACTERISTICS:

Comercial name Certrol H

Composition Ioxynil 12% ; MECOPROP 36% . Manufacture Comercial Quimica MASSO S.A Activity Herbicide

Certrol H

0,0075 7,5

0,75

0,075

0,001 0,01 0,1 1 10

0 5 10 15 20 25

Tim e in days

D o s e m g /c m

2

(l o g s c a le )

Filter paper Standard-soil

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

(72)

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.

Tordon 101

4,7

0,0047 4,7

0,047 0,47

0,001 0,01 0,1 1 10

0 5 10 15 20 25

Time in days

D os e m g/ cm

2

(lo g sc al e)

Filter paper Standard-s oil

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

(73)

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.

Lugsamag-N

3

0,3

0,03 3

0,01 0,1 1 10

0 5 10 15 20 25

Time in days Dose mg/cm2 (log scale)

Fig 3. Time and dose relatioship for wich 100% mortality was obtained.

(74)

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.

Garlon Gs

0,018

0,0018 0,18

1,8

0.18

0,001 0,01 0,1 1 10

0 5 10 15 20 25

Time in days Dose mg/cm2 (log scale)

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

(75)

CHARACTERISTICS Comercial name Chas 4

Composition Clorpirifos 48% & aroamatic hidrocarbures.

Manufacture AGRODAN S.A Activity Insecticide

Chas-48

0,07

0,7 0,7

7

0,001 0,01 0,1 1 10

0 5 10 15 20 25

Time in days

D o se m g /c m

2

( lo g s ca le )

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

(76)

1,178

0,55

0,055 0,75

0,157 0,23

0,047 0,023

0,0023 1,57

0,001 0,01 0,1 1 10 100

0 5 10 15 20 25

Time in days

D o s e m g /c m

2

(l o g s c a le )

METALDEHIDE AMIGO DIMILIN AZADIRACHTIN

NOMOLT SAPONIN BACTOSAN

Fig.11 Time and dose relatioship for wich 100% mortality was obtained.

(77)

Results on slug eggs: slurry

100 125 150 175 200 225 250 275 300

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Exposure in days LD50 in mg cm-2

pig-slurry caw-slurry

(78)

SCREENING ON PLANT EXTRACTS FOR OVICIDAL ACTIVITY AGAINST Deroceras reticulatum EGGS

University of Santiago de Compostela Faculty of Biology

Spain

José CASTILLEJO & 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”

(79)

Looking for a Biomolluscicides

PLANTS

PLANT EXTRACTS

ACTIVE PRODUCT

SYNTHETIC PRODUCT

®

PATENT COMMERCIALIZATION

HOW and WHEN to apply it ? AGAINST EGGS

AGAINST ADULTS

79

(80)

Preparing the extracts for testing:

 Plants were collected and identified

 Different parts of the plants were separated:

Rhizomes Leaves Fruits

(81)

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

(82)

Euphorbia helioscopia L.

 Common name: Sun spurge (Euforbia)

 Constituents:

euphorbone; 12-

deoxyphorbol; wax;

resin; lignin; basorrin;

volatile oils; euphorbo- resene; euphorbic acid;

calcium malate

SPURGE: Any of various euphorbiaceous plants of the genus Euphorbia that have milky sap and small flowers typically surrounded by conspicuous bracts. Some

species have purgative properties

(83)

Euphorbia’s extracts tested

LEAVES STEAMS ROOTS FRUIT

FRESH DRY FRESH DRY FRESH DRY FRESH DRY

Fp Fp Fp Fp Fp Fp Fp Fp

W A T E R

(ppm)

100,000        

80,000        

50,000        

40,000        

8,000        

ACENTO NE / WATER

(ppm)

100,000        

(84)

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

(85)

Ruta’s extracts tested

LEAVES STEAMS FLOWERS

FRESH DRY FRESH DRY FRESH DRY

Fp Ss Fp Ss Fp Ss Fp Ss Fp Ss Fp Ss

W A T E R

(ppm)

100,000

   -    -    -

80,000

           

50,000

   -    -    -

40,000

           

10,000

  - -    -   - -

8,000

           

1,000

  - -    -   - -

100

  - -    -   - -

A C E T O N E / W A T E R (ppm)

100,000

       -    

80,000

           

50,000

   -    -    

40,000

           

10,000

   -    -    -

8,000

           

1,000

  - -   - -   - -

100

  - -   - -   - -

Fp = Filter paper Ss = Standard soil

Ruta’s dry leaves

acetone / water (7:3) extracts Mortality over time

0 20 40 60 80 100

0 5 10 15 20 25

Time in days

% mortality

80000 ppm 40000 ppm 20000 ppm

10000 ppm 5000 ppm

(86)

DEFINITIVE Ruta’S EXPERIMENTS ON FILTER PAPER

LEAVES DRY

Fp

W A T E R (ppm)

80,000 

40,000 

20,000 

10,000 

5,000 

ACETONE / WATER

(ppm)

80,000 

40,000 

20,000 

10,000 

5,000 

(87)

Ruta’s dry leaves Water extracts

Mortality over time

0 20 40 60 80 100

0 5 10 15 20

Time in days

% m o rt a li ty

80000 ppm 40000 ppm 20000 ppm

(88)

Dose-response curve/ LD ₅₀

22939 18455

32707

0 20 40 60 80 100

1000 10000 100000

Dose in ppm (log scale)

% m o rt a li ty

5 days 7 days 10 days LD50

(89)

Ruta’s dry leaves

acetone / water (7:3) extracts Mortality over time

0 20 40 60 80 100

0 5 10 15 20 25

Time in days

% m o rt a li ty

80000 ppm 40000 ppm 20000 ppm

10000 ppm 5000 ppm

(90)

Dose-response curve

27841

7862 16907

0 20 40 60 80 100

1000 10000 100000

Dose in ppm (log scale)

% m o rt a li ty

2 days 3 days 5 days LD50

(91)

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

(92)

Digitalis’ extract tested

LEAVES FRESH

Fp

WATER (ppm)

80,000 

40,000 

8,000 

ACETONE / WATER

(ppm)

100,000 

Fp = Filter paper

Ss = Standard soil

(93)

(94)

(95)

Searching for Nematode Strains

(96)

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.

The Spanish Phasmarhabdities strain

(97)

97

Our strategy versus Bayer, Lonza, Ferramol stratgy

Bayer Methiocarb

Lonza

Metaldehyde Ferramol

Iron phosphate

(98)

1.4 m

2 m

²

PARCEL miniPLOTS

• Six replicates for each crops/test

• Six controls plots/random selection

• Monthly sampling

2.6 m

(99)

Periphery

(Nothing)

Centre

Phasmarhabditis Metaldehyde Ioxynil (Herbicide)

Phasmarhabditis + Ioxynil

Phasmarhabditis + Metaldehyde

14 m

²

2 m

²

Before starting experiments the soil fauna was assessed on every plot and its spatial distribution was homogeneous

AREA FOR INFAUNAE SAMPLING

(100)

100

(101)

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

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.

Eggs Eggs Eggs

Eggs

101

(102)

102 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

THE NEW PROJECT

(103)

103

• Defining and rewriting the Work Packages

(104)

104

(105)

WP1 Land snails ‘ biological cycle

WP2

Diet composition, trap plants..

WP 6 and WP 7 Fi eld experi men ts

WP4

Biomoll tests

WP5

Bioovicide Agrochem tests

WP3 Statistical models to predict activity

WP Collaboration and Dissemination

WP Project Management

W P Explo it ati on

Project

Start Project

End

Marivonne PERT

(106)

106

Dr. José Castillejo & Dr. Javier Iglesias Departamento de Zoología

Facultad de Biología

Universidad de Santiago de Compostela 15782 Santiago de Compostela La Coruña. Galicia. España Dr Maryvonne Charrier, MC

Université de Rennes 1- UMR EcoBio 6553 Campus de Beaulieu, Bât. 14A

F-35042 RENNES Cedex France

Dr. Olaf Schmidt Ph.D. (Senior Lecturer) UCD School of Agriculture, Food Science and Veterinary Medicine. University College Dublin. Belfield, Dublin 4. Ireland

Dr. Solveig Haukeland. Bioforsk Norwegian Institute for Agricultural and Environmental Research Fr. A. Dahlsvei 20, N-1432 Ås Norway (Noruega)

Dr. Grita Skujienė Department of Zoology Faculty of Natural Sciences Vilnius University Ciurlionio 21/27, 03101 Vilnius Lithuania

Dra. Mª Mercedes Ortega Hidalgo

Dpto. Gent. Antrop. Física y Fisiología Animal Facultad de Ciencia y Tecnología

Universidad del Pais Vasco. España

Dr Eva Knop University of Bern

Institute of Ecology and Evolution Baltzerstrasse 6

CH-3012 Bern. Switzerland Dr Albert Ester

PAV. Edelhertweg P.O. Box 430

8200 AK Lelystad. The Netherlands

Syngenta Crop Protection Syngenta Crop Protection UK Ltd.

CPC4 Capital Park. Fulbourn Cambridge CB21 5XE. UK

E-mail: [email protected] Dr. André Chabert

ACTA. 4 place Gensoul 69287 Lyon Cedex 12 France

Dr Georges Dussart

Canterbury Christ Church University North Holmes Road

Canterbury Kent, CT1 1QU United Kingdom

(107)

107

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

Dr. Intidhar ABBES

Biodiversité et Biologie des Populations Sciences Biologiques

Faculté des sciences de Tunis

Université Tunis El Manar, 2092, Tunisie

(108)

108

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

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. Galileo Rivas

Líder Programa de Producción Agroecológica de Cultivos Alimenticios

Centro Agronómico Tropical de Investigación y Enseñanza, CATIE División de Investigación y Desarrollo

7170 Turrialba, Cartago. Costa Rica

Dra. Lenita de Freitas Tallarico Lenita de Freitas Tallarico (Biologist) Laboratorio de

Parasitología/Malacología Instituto Butantan Avda. Voital Brasil, 1500 CEP-05503-900 Sao Paulo. Brasil

Dra. Luz Elena Velásquez Trujillo

Programa de Estudio y Control de Enfermedades Tropicales –PECET Universidad de Antioquia

Medellín, Colombia

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

(109)

109

Syngenta Crop Protection Syngenta Crop Protection UK Ltd.

CPC4 Capital Park. Fulbourn Cambridge CB21 5XE. UK

E-mail: [email protected]

(110)

WP 1

Manager: Participant ? Participants 1, 2 ,3, 4, 5, 6, 7,

8

WP ????

8

WP 7

Manager: Participant ? Participants 1,2,3,4, 5, 6, 7, 8

WP 6

8

WP 5

Manager: Participant ? Participants 1, 2 ,3, 4, 5, 6, 7.

8

WP 3

8

WP 4

8

WP 2

8

DELIVEREBLES

•Statistical model to predic activity

• Molluscicides ovicicides

• Nematode zooparasitic

• Integrate package for organic crops

• Integrate package for convencional crops

COORDINATION Participant 1

PROJECT MANAGEMENT STRUCTURE

Beware!. Coordination for each WP

(111)

WP 1

LAND SNAIL’S BIOLOGICAL CYCLE

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

WP 9

NEMATODO ZOOPARASITIC Phasmarhadities Participant: Latino America

WP 7 & 8

FIED EXPERIMENTS CONVENTIONALLY, ORGANICS CROPS

Participant 4, 5, 6, 7, 8

WP 6

PIG AND COW MANURE AS BIO OVICIDES TESTS

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

WP 5

BIO OVICIDAS AGROCHEMICALS TESTS

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

8

WP 3

STATISTICAL ACTIVITY MODEL

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

WP 4

BIOMOLLUSCICIDES TESTS COLLATERAL EFFECTS

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

WP 2

DIET COMPOSITION TRAP PLANTS

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

• Statistical model to predic activity

• Molluscicides ovicicides

• Nematode zooparasitic

• Integrate package for organic crops

• Integrate package for convencional crops

RELATIONSHIP TASKS and Participants

(112)

WP 1

LAND SNAIL’S BIOLOGICAL CYCLE

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

WP 9

NEMATODO ZOOPARASITIC Phasmarhadities Participant: Latino America

WP 7 & 8

FIED EXPERIMENTS CONVENTIONALLY, ORGANICS CROPS

Participant 4, 5, 6, 7, 8

WP 6

PIG AND COW MANURE AS BIO OVICIDES TESTS

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

WP 5

BIO OVICIDAS AGROCHEMICALS TESTS

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

8

WP 3

STATISTICAL ACTIVITY MODEL

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

WP 4

BIOMOLLUSCICIDES TESTS COLLATERAL EFFECTS

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

WP 2

DIET COMPOSITION TRAP PLANTS

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

RELATIONSHIP TASKS and Participants

intercommunication

(113)

Participant 2

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

Participant 8

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

Participant 7

Participant 6

Participant 5

Participant 4

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

Participant 4

Participant 3

Participant 1

Coordinator USC Spain

WP. 1, 2 ,3, 4, 5, 6, 7, 8

RELATIONSHIP Participants and TASKS

(114)

Participant 10

Participant 9

Participant 6

Participant 5

Participant 4

Participant 2

Participant 3

Participant 1

RELATIONSHIP Participants and TASKS

Participant 8

Participant 7

intercommunication

(115)

Participant 10

Participant 9

Participant 6

Participant 5

Participant 4

Participant 2

Participant 3

Participant 1

Participant 8

Participant 7

Syngente

Provide non residual

agrochemicals for testing

(116)

116

Work Packge Nº

Task

Nº Work Packages Title

Hours per day

Number of days per month

Number of month

Number of person participating

Total Hours per WP or Task

Person- month Hours/135 WP.1

Land Snail Biological Cycle.

the size, structure and dynamics of their

populations 8 5 24 2 1920 14.2

WP.2.

Land Snails diet composition. Trap plants

12 5 24 2 2880 21.3

WP.3.

Statistical Model to predict land snails activity

12 5 24 2 2880 21.3

WP.4.

Task 4.1.

Bio pesticides, Bio Molluscicides, Bio Ovicides Plants Extracts. Laboratory test on paper filter

Standard soil 4 10 30 2 2400 17.7

Task 4.2.

Mini plots tests on horticultural soil

2 10 24 2 960 7.1

Task 4.3. Collateral effects on soil fauna

2 10 18 2 720 5.3

Task 4.4.

Chemicals analysis to search the active principle of plant extracts with mulliscicide and ovicide

activity 2 10 18 2 720 5.3

WP.5.

Task 5.1.

Laboratory test to find non residual

agrochemicals with ovicidal activity 4 10 25 2 1920 14.2

Task 5.2.

Field experiments on conventionally horticultural crops to evaluate the efficacy of selected agrochemical as

molluscicide-ovicide

4 10 18 2 1440 10.6

Task 5.3.

Field trials to evaluate the collateral effects on soil Fauna and border effect on wild land snails

of ovicide agrochemicals. 1 10 18 2 360 2.6

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

For carrying out the project. People involved:

• 1 person full time dedication. PhD student + 2 person partial time dedication. Staff University

(117)

117

WP.6.

Task 6.1. Manure Laboratory test on land snail eggs

for accurate Ovicidal concentration 2 10 18 2 720 5.3

Task 6.2.

Field experiments to evaluate the efficacy of cow and pig manure as slug eggs control for key organic horticultural crops

2 10 24 2 960 7.1

Task 6.3.

Field analysis to investigate the collateral effect on soil fauna and border effect on wild land snails of cow and pig manure

1 10 18 2 360 2.6

Task 6.4

Field experiments to use tramp-plants as deterrent method to protect organic horticultural crops alone and in combination of cow and pig manure

1 5 24 2 240 1,7

WP. 7

Field experiments in conventionally crops to evaluate the efficacy of ovicide agrochemicals alone and in combination with other commercial molluscicides

4 10 24 2 1920 14.2

WP. 8

Field experiments in organics horticultural crops to evaluate the efficacy of organic molluscicides-ovicides and the use of plant-traps

4 10 24 2 1920 14.2

WP. 9

To identify improved strains of

Phasmarhabditis nematodes which are more effective biocontrol agents of larger slug species in Hispano-America.

1 10 36 2 720 5.3

TOTAL person-months 209.7

(118)

118

For carrying out the project. People involved:

• 1 person full time dedication. PhD student

• 2 person partial time dedication. Staff University

(119)

119

Work Packge Nº

Task

Nº Work Packages Title

Type activity

Lead participant

nº

Lead Participant short name

Total Person- months

Start month End month

WP.1

Land Snail Biological Cycle.

the size, structure and dynamics of their

populations RTD 1 24

WP.2.

Land Snails diet composition. Trap plants

RTD 1 24

WP.3.

Statistical Model to predict land snails

activity RTD 13 36

WP.4.

Task 4.1.

Bio pesticides, Bio Molluscicides, Bio Ovicides

Plants Extracts. Laboratory test on paper filter

Standard soil

RTD 1 30

Task 4.2.

Mini plots tests on horticultural soil

RTD 13 36

Task 4.3.

Collateral effects on soil fauna

RTD 19 36

Task 4.4.

Chemicals analysis to search the active principle of

plant extracts with mulliscicide and ovicide activity

RTD 19 36

WP.5.

Task 5.1.

Laboratory test to find non residual agrochemicals

with ovicidal activity RTD 1 24

Task 5.2.

Field experiments on conventionally horticultural crops to evaluate the efficacy of selected agrochemical as

molluscicide-ovicide

RTD 13 30

Task 5.3.

Field trials to evaluate the collateral effects on soil

Fauna and border effect on wild land snails of

ovicide agrochemicals.

RTD 19 36

Gantt Chart showing the timing of the different WPs and their components.

(120)

120