Work Package 2 - Universidade de Santiago de Compostela

CALL: FP7-KBBE-2011-4
Proposal full title: Novel strategies for integrating land snail pests control of
agricultural crops in Europe, with projection to Latin-
America
Proposal acronym:
Short name:
Type of funding scheme:
Work programme topics addressed:
LAND SNAIL PEST CONTROL
Land snail’s life cycle as pest control core
Collaborative Project. CALL: FP7-KBBE-2011-4
Food, Agriculture and Fisheries, and Biotechnology
Area: 2.1.2
Topic number: KBBE.2011.12-05
Name of the coordinating person: Dr. José Castillejo Murillo
Departamento de Zoología y Antropología Física. Facultad de Biología.
Universidad de Santiago de Compostela. E-15782 Santiago de
Compostela. La Coruña. Galicia. España.
Mobile Phone: + 34 654 969 784
Tel: + 34 981563100
Fax: + 34 981 596904. E-mail: jose.castillejo@usc.es
List of participants
WORK PROGRAMME 2011
COOPERATION
THEME 2
FOOD, AGRICULTURE AND FISHERIES, AND BIOTECHNOLOGY
(European Commission C(2010)4900 of 19 July 2010)
Participant number Participant organitation name Country
1 (Coordinator) Universidad de Santiago de Compostela Spain
2 Gordon Port School of Biology, Newcastle University, UK UK
3 Georges Dussart Canterbury Christ Church University, Kent UK
4 Marivonne Université de Rennes 1-UMR EcoBio 6553 France
5 Rita Triebskorn Physiologische Ökologie der Tiere Germany
6 Solveig Bioforsk Norway
7 Grita Faculty of Natural Sciences. Vilnius University Lithuania
8 Eva Knop Institute of Ecology and Evolution Switzerland
9 Albert Esther LEI Detachement Lelystad The Nedherland
10 Letelier Museo Nacional de Historia Natural. Santiago de Chile Chile
11 Salvio Faculty of Agricultural Sciences. University of Mar del Plata Argentina
12 Lenita Instituto Butantan Brazil
13 Trujillo Universidad de Antioquia Colombia
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15
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Novel strategies for integrating land-snail pest control of agricultural crops in
Europe, with projection to Latin-America
TITLE: Novel strategies for integrating land-snail pests control of agricultural crops in Europe, with projection to
Latin-America.
1: Scientific and technical quality, relevant to the topics addressed by the call
1.1 Concept and objectives
To introduce in a series of traditional and ecological 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 ovicidal molluscicides
(to destroy the egg lays), when to apply the biological control through parasite nematodes or when to use the
trap-plants; and all this strategy will be regulated by the statistical model witch predict the activity. To develop
this integrated control methods it will be necessary:
1. To know the biotic and a biotic factor that describe the land snails biological cycle in different areas of
Europe and Latin-America, to elaborate the control method.
2. To know the specific land snails´ diet with the objective of finding the most attractive plant species to use
them as trap-plants.
3. To understand the activity in function of the environment biotic and a biotic variables, with the aiming of
developing an effective abundance and activity statistical prediction method.
4. To search plants with bio pesticide activity for be used as bio-molluscicides and bio-ovicides against land
snail and its eggs.
5. To know the ovicidal potential of the usual non residual agrochemicals to use them as molluscicidesovicides
in crops.
6. To know the ovicidal potential of cattle’s and swine’s slurries as molluscicides-ovicides in ecological
farming.
7. To search in each study areas of Latin-America a parasite nematode (Phasmarhabdities alike) to use it as a
biological control method against land snails.
8. To deliver to the horticultural industry an effective integrated crop management strategies, low chemical
and biological protecting methods against land snails.
9. To deliver to the ecological farming effective integrated packages methods based on cattle and swine
manure and plant traps, to protect the crops against land snail pests.
With these strategies we are settling the basis for an integrated control method, to achieve a more rentable
crop due to; having less plant damages, less pesticides use, and a more respectful farming with the environment
and the wild fauna. The land snail pest problem in Europe and Latin-America is increasing every day due the
commercial globalization. The most dangerous land snail species in Europe are: Arion lusitancicus, Deroceras
retiuclatum, Lehamnnia marginata, Milax gagates, Criptophalus aspersus and Theba pisana among others. The
90% of the land snail species that are pest in Latin-America are introduced species from Europe and other
countries, this species proliferate indiscriminately because of the absence of natural predators, such as Acanthina
fúlica, Deroceras reticulatum, Milax gagates, Lhemannia marginata, Arion intermedius, Criptophalus aspersus….
In some Caribbean regions native land snail species cause important damages, such is the case of Veronicella
genus in Mexico and Cuba.
1

Scientific and technical objectives detailed description
This project requires the application of novel control strategies to control native or introduced land snail
pests in crops that can be used in any agricultural pest. With the strategies proposed here we can anticipate to
the damages caused by the land snail pests, due the application of a preventive method even before damages
appear on the crops.
Our strategies are based on:
1. To understand the land snails biological cycle in the crops study areas.
2. To understand the land snails activity in function of the climatic variables and the crop type.
3. To destroy the land snail´s egg-lays thanks to the plant extracts and non residual standard
agrochemicals collateral effect.
4. To rationalize standard molluscicides consumption in standard farming.
5. To introduce cattle and swine manure and trap-plants as control methods in organic farming.
6. To understand the collateral effects of the products used in this integrated pest control methods.
The understanding of the biological cycle is very important, because we attempt to use the molluscicides
before damages appear. With the knowledge of the biological cycle we will know which time of the year is
juvenile, adult or senile, we will know when the egg-lays are done, and in other words, we will know the sizes,
structure and dynamics of their populations. The information provided by the biological cycle is important to
implement this new strategy, as these molluscicides must be applied when the population density is lower and
when there are fewer egg-lays in the soil. With this we obtain an optimal effectiveness destroying the egg-lays
through ovicidal and also killing gastropods. By applying less molluscicides we save money and minimize the side
effects on the environment.
Knowing the diet of land snails in the study areas will give us information on the possible use of trap-plants,
to evaluate and estimate the damage that they actually produce on crops. By studying the stomach contents of a
specified number of land snails we can know their preferences, in previous research we found that plants that
had a low abundance in the environment, appeared with high frequency in the stomach of land snails, this means
that they have positive selection for this type of plants. These plants can be used as trap-plants to protect
vegetable crops deterring land snails to eat the trap-plants. It is a very useful strategy in organic farming.
Knowing the activity of terrestrial gastropods in terms of climatic variables and crop phenology is required
to develop a statistical for activity prediction. With this model we can predict with 24-48 beforehand the activity,
and provide the farmer information that land snails will be active, and thus may apply the traditional
molluscicides at the right time, getting a greater effectiveness using smaller amount of molluscicides, leading to
saving resources and reducing side effects on plants, soil and wildlife.
So far we have been talking about traditional molluscicides. The tradiotional molluscicides (metaldehyde,
carbamate, iron sulphate, phasmarhadities...) are intended to kill the individuals, in other words, kill the
terrestrial gastropods leaving intact the land snail´s egg-lays in the soil.
There is a growing body of evidence to suggest that in the past 4-5 decades there has been an excessive
dumping of chemical toxins on the soil. As a result the soil has become barren and ground water toxic, in many
places. Contrast this with organic inputs that are safe, non toxic, and cost much less. 'Biopesticides' are certain
types of pesticides derived from such natural materials as animals, plants, bacteria, and certain minerals. Benefits
of biopesticides include effective control of insects, plant diseases and weeds, as well as human and
environmental safety. Biopesticides also play an important role in providing pest management tools in areas
where pesticide resistance, niche markets and environmental concerns limit the use of chemical pesticide
products. To find plants extract with molluscicidal and ovicidal activity against land snails and their eggs are
original and promising.
In each country, agricultural authorities allow a number of non residual agrochemicals for different uses
and for different purposes, can be fertilizers, herbicides, acaricides, fungicides, etc... These products have gone
through a series of tests to be authorized. In previous research we have tested non residual agrochemicals to see
if they had the potential to destroy the land snail´s egg-lays, and many of them had as side effect the egg-lays
killing. Basing on these assumptions each partner must make a test series with non residual agrochemicals
approved in their country, to discover which one or ones have a higher ovicidal power at the lowest concentration
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in the shortest time. In organic farming the use of synthetic chemicals such as fertilizers, pesticides, antibiotics,
etc.., it´s forbidden, with the objective of preserving the environment, maintaining or enhancing soil fertility and
provide food with all its natural properties. Fertilizers that can be used in these kinds of crops can be of two types,
green fertilizers or livestock manure. In previous research we found that certain concentration of swine and cattle
manure had ovicidal action on terrestrial gastropod egg-lays. Therefore each partner will have to do tests to
discover the type and concentration of manure that has higher ovicidal power against the land snail´s egg-lays in
their study areas.
Finding a new parasite-nematode with a biological cycle like Phasmorhadities hermaphrodita is crucial to
have a new tool for biological land snail pests control in agriculture in Latin-America, as the European variety has
problems at certain soil temperatures. The task to find European zoo parasitic nematode was made in previous
97- UE - Project.
As a consequence the of traditional and organic farmers are going to get a series of useful tools for land
snail pests and, first they are going to have a new control strategy based on the application of the ovicidalmolluscicides
at the correct time, determined by the life cycle of the terrestrial gastropods, and not by the crop
phenology. It will be explained which chemicals they need to destroy land snail´s egg-lays. Through the predictive
model, the farmers will have the information necessary to know which day and a time terrestrial gastropods will
be active, thus apply the traditional molluscicides at the right, time, place and amount. This information must be
transmitted through scientific meetings, counselling State Agricultural Agencies and through web pages of the
State Servers.
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Compliance with the objectives of the work programme and its priorities
This project can definitely solve the problem of land snail pests in agriculture that has been globalized by the
reform of the European Common Agricultural Policy and which it is forced to fulfil in the countries from which we
import agricultural products.
This project is related to THEME 2 (Food, Agriculture and Fisheries, and Biotechnology), Area 2.1.2
KBBE.2010.1.2-05 "Integrated pest management in farming systems of major importance for Europe" out of the
7th Framework Program of Cooperation (FP7 Cooperation Work Program), and our project fits in perfectly in the
topics (issues) of integrated pest control (management) (In the context of Integrated Pest Management - IPM-) in
particular (specifically) can be said that:
1. It includes preventive measures such as, molluscicides application in the more labile phases of the life cycle
of terrestrial gastropods and when there is less density of population and eggs in the soil, which generally
coincides with (periods in which) (times when) there is nothing planted on farms .
2. The design of this strategy is based on the study of the biological cycle of pests and the study of their activity
in terms of biotic and abiotic variables of the environment, representing perfect control and more accurate
information to take preventive measures.
3. With this strategy control measures are applied at the right time, anticipating the emergence of the pest,
which implies less molluscicide applied to achieve a better effect, besides the molluscicide is never next to
plants, if the measures control are applied prior to planting. Our strategy is aimed at controlling or
eradicating the pest to protect the crop, in other words, we use a preventive strategy.
4. It is an integrated control based on the decision-making through the statistical model prediction. We only
use low toxicity chemicals, we use the biological control of nematodes through zooparasites, not to mention
the use of trap plants to deter terrestrial gastropods that attack crops or the use of swine and cattle manure
as ovicidal all this applied at the time we enter the life cycle of the pest snail and the predictive model.
5. Con este proyecto estamos sentando las bases para que los agricultores cumplan la Directive 2009/128/EC of
The European Parliament and of the Council of 21 October 2009 establishing a framework for Community action to
achieve the sustainable use of pesticides, según la cual by 14 December 2018 will be necessary to minimise the
hazards and risks to health and environment from the use of pesticides. Al finalizar este proyecto dispondremos de biomolusquicidas
with molluscicida and ovicidal action. Syngenta and Bayer Companies are intrested in this WP.
6. 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.
7. This project combines "combine modelling and experimentation" because the entire strategy is based on
the study of biological and ecological cycle of the pest, its dynamics, in order to achieve the greatest success
of control with the least effort and with the least means.
8. The risks of not succeeding in this project are limited, first all the partners who are part of the research
group have demonstrated ability to perform all the tasks outlined in the Work Packages, and also the USC
team that coordinates this project has an extensive experience in developing predictive models of activity of
the snails and slugs in agricultural crops of Galicia (Spain) and in controlling pests in vegetable crops and
vineyards and many others European partners have similar experience in similar fields. Given the
globalization of trade, over 90% of land snail species pests in Latin-America are of European origin, in other
words, they are introduced species that we have been working with over 20 years.
9. The balance between costs and benefits will always be positive because we will control the final shape of
land snail pests in agriculture, and we will bring to the traditional and organic farmers to have a number of
tools to obtain a more profitable crop and seeding time management, very respectful with the environment.
10. Finally, all information obtained from this project and the control strategies are available for the farmer,
either through meetings, workshops taught by the competent authorities or available through “on line”
services were the farmers will resolve questions and provide information of the pest activity.
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1.2 Progress beyond the state-of-the-art
A escala mundial, los perjuicios económicos causados por los gasterópodos terrestres se han
incrementado gracias a la globalización del comercio lo que conlleva que se hable de especies de caracoles y
babosas introducidas de un continente a otro, especies que al no tener depredadores específicos presenta un
crecimiento poblacional exponencial. Mientras que algunos caracoles terrestres pueden alcanzar el estatus de
plaga incluso en regiones relativamente áridas, las babosas resultan especialmente problemáticas en climas
templados y lluviosos, pero aún en este caso, la magnitud de los daños causados por las babosas a los cultivos
varía mucho a escala regional y de un año a otro (Port y Port, 1986). Muchos especialistas coinciden en señalar
que los daños ocasionados por los gasterópodos se han incrementado de forma muy significativa en las últimas 2
ó 3 décadas, debido a la conjunción de una serie de factores como la simplificación de las técnicas de cultivo
(reducción del laboreo, siembra directa), la reducción de las poblaciones de insectos depredadores por el uso de
insecticidas, o la utilización de nuevas variedades de cultivo más susceptibles al ataque de los gasterópodos
(Hommay, 1995, 2002; Godan, 1999; Speiser, 2002). Por otro lado, la elevación de los estándares de calidad
exigidos por los consumidores hace que la tolerancia del mercado a productos dañados sea cada vez menor, lo
que se traduce en una intensificación de las medidas de control de plagas.
In recent years, the problems caused by land snails, especially the grey field slug (Deroceras reirulatum),
the Spanish slug (Arion lusitanicus), the brown garden snail, Cryptomphalus (Helix = Cantareus) asperses,the
white garden snail, Theba pisana 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)
Land snails reduce the vigour of some crops by killing seeds or seedling, by desroying stems or growing
points, or by reducing the leaf area. This may slow down crop development and /or reduce yield. In other crops,
the harvest is devaluated by feeding damage, mucus trails, faeces or presence of land snails. Land snail feeding
may also initiate mould growth or rotting. Damage by land snails in not always easily distinguished from insect
feeding. Clear, silvery mucus trails indicate land snail activity.
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).
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 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.
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).
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El control de plagas de gasterópodos terrestres se realiza, de forma casi exclusiva, por medio de la
aplicación de cebos (“pellets”) que contienen entre un 2% y un 8% de metaldehído o de carbamatos (Godan,
1983, 1999; South, 1992; Garthwaite y Thomas, 1996; Bailey, 2002; Speiser, 2002). El principal productor mundial
de metaldehído es la empresa suiza Lonza. El carbamato más utilizado en el control de plagas de gasterópodos
terrestres es el metiocarbamato, cuya licencia de fabricación es propiedad de la empresa alemana Bayer. Ambos
compuestos muestran una eficacia similar en lo que se refiere a su capacidad para reducir los daños causados por
los gasterópodos a las plantas (Bailey, 2002), y también ambos presentan efectos negativos sobre las poblaciones
de otros grupos de animales (South, 1992; Bailey, 2002). Buchs, Heimbach y Czarnecki (1989) han señalado la
existencia de efectos negativos de los cebos molusquicidas con metaldehído sobre las poblaciones de algunos
carábidos, y Bieri, Schweizer, Christensen y Daniel (1989) han documentado una reducción de la abundancia de
carábidos y estafilínidos tras la aplicación de cebos molusquicidas con metiocarbamato en praderas. Aunque en la
actualidad todos los cebos molusquicidas incorporan pigmentos (generalmente azules) y otras sustancias para
reducir el riesgo de ingestión por parte de mamíferos y aves, son frecuentes los casos de envenenamiento de
animales domésticos debido al consumo de cebos molusquicidas (Bailey, 2002). A finales de los años 80, los cebos
molusquicidas con carbamatos fueron prohibidos en muchos estados de Norteamérica, debido a la elevada
frecuencia de casos de envenenamiento de aves que se registraron (Sakovich, 1996). Tarrant y Westlake (1988)
señalan que la utilización de cebos molusquicidas con metiocarbamato supone una seria amenaza para las
poblaciones del ratón de campo, Apodemus sylvaticus (Linnaeus, 1758). Keymer, Gibson y Reynolds (1991)
registraron elevadas concentraciones de acetaldehído (resultante de la despolimerización del metaldehído en el
tubo digestivo) en erizos (Erinaceus europaeus (Linnaeus, 1758)) encontrados muertos en el campo, y Gemmeke
(1997) observó síntomas de envenenamiento y casos de fallecimiento, en erizos alimentados con babosas que
habían ingerido cebos con metiocarbamato.
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
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.
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.
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Directive 2009/128/EC of The European Parliament and of the Council of 21 October 2009 establishing a
framework for Community action to achieve the 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.
En los últimos años ha aparecido en el mercado un nuevo molusquicida químico, bajo el nombre comercial
de Ferramol , fabricado por la empresa alemana Neudorff GMBH. Este producto se presenta también en forma
de cebos y contiene fosfato de hierro como ingrediente activo. Los ensayos realizados hasta la actualidad para
comprobar su eficacia (Iglesias y Speiser 2001; Speiser y Kistler, 2002) indican que ésta es equiparable a la de los
molusquicidas químicos clásicos, metaldehído y metiocarbamato. Sin embargo, a diferencia de éstos, que son
totalmente sintéticos, el fosfato de hierro aparece de forma natural formando parte de varios minerales,
especialmente la strengita (Fe III PO4 2(H2O) ortorrómbico) y metastrengita (Fe III PO4 2(H2O) monocíclico) (Roberts,
Campbell y Rapp, 1990; Clark, 1993), y es un compuesto con una toxicidad muy baja (EPA, 1998).
La falta de medios de control de plagas de gasterópodos cuya utilización esté autorizada en la agricultura
biológica hace que estos animales hayan sido considerados como los más dañinos para los cultivos biológicos por
numerosas asociaciones profesionales de Gran Bretaña y Suiza (Peackock y Norton, 1990; Kesper y Imhof, 1998).
El único agente de control biológico que se comercializa en la actualidad para el control de plagas de babosas es
el nematodo Phasmarhabditis hermaphrodita (Schneider, 1859), lanzado al mercado por primera vez en Gran
Bretaña en 1994. Numerosos ensayos de campo realizados en una amplia variedad de cultivos y de países
europeos han puesto de manifiesto que P. hermaphrodita es capaz de reducir los daños ocasionados por las
babosas a las plantas (Wilson, Glen y George, 1993; Wilson, Glen, George y Hughes, 1995; Wilson, Hughes y Glen,
1995; Ester & Geelen, 1996; Iglesias, Castillejo & Castro, 2001ab). Su eficacia frente a la especie D. reticulatum
está fuera de toda duda (Glen, Wilson, Brain y Stroud, 2000), pero existen indicios de que su eficacia contra otras
especies podría ser menor (Wilson et al., 1995a; Coupland, 1995; Glen et al., 1996; Speiser y Andermatt, 1996;
Speiser, Zaller y Neudecker, 2001; Iglesias y Speiser, 2001). La eficacia de los tratamientos con P. hermaphrodita
está muy condicionada por la temperatura y la humedad del suelo, que afectan en gran medida a su
supervivencia, pero presenta la ventaja de que las condiciones de temperatura y de humedad que son favorables
para la actividad de las babosas lo son también para la supervivencia del nematodo, mientras que los
molusquicidas químicos en forma de cebos ven muy mermada su eficacia en las condiciones de elevada humedad
en las que los gasterópodos ocasionan la mayoría de los daños a las plantas (Glen et al., 1996). No obstante, el
elevado coste económico que suponen en la actualidad los tratamientos de control de plagas con nematodos
7
The figure examines 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.

hace que su uso esté todavía muy restringido a cultivos de elevado valor como las plantas ornamentales y algunas
hortalizas (Grunder, 2000).
La aplicación de molusquicidas representa sólo una medida de control a corto plazo, es decir, con ellos se
consigue proteger temporalmente a las plantas de los daños que podrían causarle los gasterópodos. Sin embargo,
no tienen un efecto significativo y duradero sobre las poblaciones de gasterópodos residentes en las zonas de
cultivo, por lo que el riesgo de que produzcan daños es permanente (Hommay, 2002; Port y Ester, 2002). Ello se
debe a que los molusquicidas aplicados afectan sólo a una parte de la población, y a que los huevos de los
gasterópodos, que se encuentran en el suelo, no se ven afectados por los tratamientos molusquicidas
convencionales, dando lugar a una rápida recuperación de las poblaciones (Glen, Wiltshire y Milson, 1988). Se ha
estimado que los tratamientos molusquicidas a base de cebos con metaldehído o carbamatos matan a menos del
50% de la población de gasterópodos existente en el momento de la aplicación (Glen y Wiltshire, 1986; Wiltshire y
Glen, 1989; Glen, Wiltshire y Butler, 1991). Por otro lado, es frecuente que la cantidad de cebo molusquicida
ingerido por los gasterópodos en el campo tenga sólo un efecto subletal transitorio (Kemp y Newell, 1985;
Wedgwood y Bailey, 1986; Briggs y Henderson, 1987; Bourne, Jones y Bowen, 1988), y se ha comprobado que la
fecundidad de los individuos que experimentan ese tipo de envenenamiento subletal no se ve afectada, por lo
que continúan poniendo huevos una vez que se recuperan (Kemp y Newell, 1985).
En los años 60 surge el concepto del control integrado de plagas (CIP) (Stern, Smith, van der Bosch y
Hagen, 1959), que en la actualidad es parte integrante de otro concepto, más amplio, que es el del desarrollo
sostenible. El control integrado de plagas implica la integración de los conocimientos provenientes de multitud de
campos (biología, química, agronomía, climatología, economía, etc.) con el fin de desarrollar las estrategias de
control más adecuadas desde el punto de vista económico, ambiental y de salud pública (Dent, 1991). Si bien es
un sistema basado en la combinación de diferentes métodos con el fin de minimizar el uso de pesticidas químicos,
no se descarta, a priori, la utilización de ningún tipo de agente de control (Coombs y Hall, 1998).
Metodológicamente, el control integrado de plagas puede describirse como un "proceso de toma de
decisiones" es el que, sobre la base de toda la información relevante disponible, hay que decidir qué medidas
tomar y en qué momento aplicarlas, para que el control de la plaga resulte, además de eficaz, lo más rentable
posible desde el punto de vista económico y lo menos agresivo que sea posible desde el punto de vista ambiental
(Bechinski, Mahler y Homan, 2002).
En la actualidad, los programas de control integrado de numerosas especies de artrópodos y de hongos
causantes de plagas en una gran variedad de cultivos, se basan en la utilización de sistemas de predicción (Dent,
1991; Frahm, Johnen y Volk, 1996). Prever en qué momento una plaga puede producir daños significativos en un
cultivo es fundamental para poder tomar una decisión con respecto a la necesidad de aplicar pesticidas para
protegerlo (Buhler, 1996). Por otro lado, dependiendo del modo de acción del pesticida, su eficacia puede estar
condicionada por la fase del ciclo en la que se encuentren los organismos causante de la plaga o por su nivel de
actividad (Bailey, 2002). En definitiva, se necesita disponer de criterios que permitan determinar tanto la
necesidad y como la conveniencia de la aplicación de pesticidas.
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
Biopesticides : an Economic Approach for Pest Management. It is hearting to observe the growing awareness
among the farmers and policy makers about ecologically sustainable methods of pest management. More and
more farmers are coming to realize the short-term benefits and long-term positive effects of the use of bioagents
and other ecologically safe methods to tackle pests. The present article 'Biopesticides' is of much relevance in this
context.
8

'Biopesticides' are certain types of pesticides derived from such natural materials as animals, plants, bacteria, and
certain minerals. Benefits of biopesticides include effective control of insects, plant diseases and weeds, as well as
human and environmental safety. Biopesticides also play an important role in providing pest management tools in
areas where pesticide resistance, niche markets and environmental concerns limit the use of chemical pesticide
products.
Biopesticides in general-
(a) have a narrow target range and a very specific mode of action.
(b) are slow acting.
(c) have relatively critical application times.
(d) suppress, rather than eliminate, a pest population.
(e) have limited field persistence and a short shelf life.
(f) are safer to humans and the environment than conventional pesticide.
(g) present no residue problems.
Pesticide residues in agricultural commodities are being the issue of major concern besides their harmful effect
upon human life, wild life and other flora and fauna.
Equally worrying thing is about development of resistance in pest to pesticides. The only solution of all these is
use of 'Biopesticide' that can reduce pesticide risks, as-
(a) Biopesticides are best alternatives to conventional pesticides and usually inherently less toxic than
conventional pesticides
(b) Biopesticides generally affect only the target pest and closely related organisms, in contract to broad
spectrum, conventional pesticides that may affect organisms as rent as birds, insects, and mammals
(c) Biopesticides often are effective in very small quantities and often decompose quickly, thereby resulting in
lower exposures and largely avoiding the pollution problems caused by conventional pesticides
(d) When used as a fundamental component of Integrated Pest Management (IPM) programs, biopesticides can
greatly decrease the use of conventional pesticides, while crop yields remain high
(e) Amenable to small-scale, local production in developing countries and products available in small, niche
markets that are typically unaddressed by large agrochemical companies.
The advances that the proposed project and patents will bring.
To this day, non residual agrochemicals with ovicidal action had not been used to control land snail pests in
agriculture. The strategy proposed in this project is innovative, original and this pest control methods are very
efficient, and makes it very easy to eradicate pest. It is original because it attempts to control the causative agent
pest in the phase of their life cycle [biological cycle] where they are most fragile, in the egg stage. It is an original
method that will develop statistical models to predict the abundance and activity of the land snail with which the
farmer can act in a preventive manner so as not to damage because it will indicate the moment when which has
to apply the ovicidal or the traditional molluscicides. It is original because it doesn´t introduce new pesticides in
agriculture, it is based in non residual agrochemicals that the farmer habitually uses, it seeks the best profile to
exploit its ovicidal activity. It is new because it puts into the hands of the organic farming a number of tools to
control land snails pests using natural fertilizers or deterrent strategies implemented by trap plants deterring land
snails from attacking crops.
It is a project that uses current technologies to see how the pests live, which are their weaknesses, and attack
them manipulating it that way to minimize side effects on crops, and even on the man. With this project we are
going to obtain results that will be patented. As a result we will be able to patent the pest control strategies,
establish a patent for the active use of non residual agrochemicals with ovicidal action against the land snail´s
egg-lays, and finally be able to patent the Statistical Prediction Activity Model and abundance of land snail that
are pests, and most likely be able to patent the use of a zooparasite nematodes for the land snail pests biological
control.
9

1.3 S/T methodology and associated work plan
1.3.1 Overall strategy of the work plan (1 page). Detailed description of the proposed work
The whole strategy is designed to perform integrated land snail pests control in conventional and organic
farming, and it is here to introduce the use of new molluscicides ovicidal action, capable of destroying the egglays,
the use of trap-plants as a deterrent, and the rational use of traditional molluscicides.
WP. 1 .-To understand the biology of land snail that are pests in a range of vegetable crops, it is necessary to
have information of the size, structure and dynamics of their populations.
WP. 2 .-To study food and qualitative and quantitative composition of the diet of terrestrial gastropods that
are pests in order to find plants that can be used as trap plants in organic farming and traditional.
WP. 3 .-To develop a statistical model that could explain and predict the abundance and activity of the land
snail as a function of environmental variables such biotic and abiotic.
WP. 4 .-To investigate the feasibility of using plant extracts as biomolluscicides and bio-ovicides. To carry this
out it will also be necessary:
4.1. Laboratory tests on filter paper (direct contact) and artificial soil (standard soil) to select the plant
extracts with molluscicidal and /or ovicidal activity.
4.2. Mini plots experiments on horticultural soil to evaluate the efficacy of the selected plant stract
against lands snails and its eggs.
4.3. Mini plots analysis to know the collateral effect of plant extract selects on invertebrate soil.
4.4. Chemical analysis to find the plant extracts active principle by analytic steps.
WP. 5 .-To investigate the feasibility of using commercial agrochemical activity as ovicidals with control land
snail egg-lays for key Conventionally grown horticultural crops. To carry this out it will also be necessary:
5.1. Laboratory tests on filter paper (direct contact) and artificial soil (standard soil) to select the
agrochemical which best suits the egg types of the pest species and soil type in the study area.
5.2. Field experiments on horticultural crops to evaluate the efficacy of the selected agrochemical as
Ovicidal-molluscicides for the pest control of key conventional grown horticultural crops.
5.3. Field analysis to know the collateral effect of agrochemical selects on invertebrate soil fauna and
border effect on wild land snails in conventional horticultural crops.
WP. 6 .- To investigate the feasibility of using swine and cattle manure of killing land snail eggs and plant-tramp
strategy of land snail pest control for key organic horticultural crops grown. To carry this out it will also be
necessary:
6.1. Laboratory testing to determine concentrations of swine and cattle manure that are effective against
the egg-lays of terrestrial gastropods pest. Discriminant trials will be made on filter paper (direct contact) and
artificial soil.
6.2. Field experiments to evaluate the efficacy of swine and cattle manure land snail egg-lays as control
for key organic horticultural crops.
6.3. Field analysis to investigate the collateral effect of swine and cattle manure on soil invertebrate fauna
and border effect on wild land snails in organic horticultural crops.
6.4. Field experiments to use plants as tramp-deterrent method to protect organic horticultural crops
WP. 7 .- Make field trials in traditional crops to compare the effectiveness of the control strategy of pest land
snail proposed by us versus conventional approaches of applying chemical molluscicides when observed damage
to the crops. Field experiments in Conventional key horticultural crops to evaluate the efficacy of selected
agrochemical ovicidal with activity against land snail egg-lays in relation to other standard commercial lowchemical
methods of killing animals land snail. Final trial.
WP. 8 .- Field experiments organics in key horticultural crops to evaluate the efficacy of organic Molluscicidesovicides
and the use of plant-traps as land snails method to control pests.
WP. 9 .- To identify improved strains of nematodes Phasmarhabditis Which are more effective biocontrol
agents of larger land snail species in Hispano-America.
10

METHODOLOGY AND RESEARCH WORK PACKAGES
Work Package 1
Field research to investigate life cycle of land snail past in horticultural crops
Partners 1 (……. Man-month)
Partners 2 (……. Man-month)
Partners 3 (……. Man-month)
Partners 4 (……. Man-month)
Partners 5 (……. Man-month)
Partners 6 (……. Man-month)
Partners 7 (……. Man-month)
Partners 8 (……. Man-month)
Partners 9 (……. Man-month)
OBJETIVES
To investigate size, structure and dynamic of land snail for a key horticultural crops
Estudiar el tamaño, estructura y dinámica de sus poblaciones.
BACKGROUND
Methodological review. Many methods have been used to make quantitative studies of land snail populations.
According to South (1992) these methods can be qualified in three categories:
A. Absolute methods, expresses the number of individuals per unit area.
B. Relative methods, expresses the number of individuals per unit effort or the relation to non-standardized traps.
C. Indirect methods, expresses sizes of population in terms of traces left or the effects produced by land snails
(for example, depending on the damage extension done to the crop, or according to bait consumption).
Relative methods are faster and much more comfortable but they have the disadvantage that the estimates are
highly dependent of the land snail activity levels, this can lead to incorrect population sizes because of weather
conditions that have an important effect on the land snail activity (Getz, 1959; Hunter, 1968a; South, 1992).
Absolute Methods involve the absolute quantification of individuals per surface area. This can be done on site by
applying an irritant substance to a determined area. For years formaldehyde was used for this purpose, but South
(1964) rejected this method when he realized that most of the land snails died before they could reach the
surface. Högger (1993) proposed a new method. First he determines an area, limiting it with a metal ring of 15 cm
of height; then he introduces it in to the ground to a depth of 5 cm. Once done this, he applies mustard oil to the
ground and captures the land snails that come out to the surface. Another method, proposed by Ferguson,
Barratt & Jones (1989), it is based on the placement of shelter traps located inside of the area determined by the
metal ring, which in this occasion is covered with a top that prevents the escape of land snails and helps to keep
the humidity (moisture) inside. The shelter traps and the area inside the ring are inspected each day removing the
land snails caught, until no new individuals appear.
Another way of obtaining absolute estimates is to take soil samples of known surfaces and transfer them to the
laboratory for further land snail extraction and quantification. The extraction can be done by; the progressive
flooding of the soil samples to make land snails surface, or by washing the soil sample on sieves with water. South
(1964) compared the last sieving method with absolute methods (flooding with cold or hot water, extraction with
chemicals, dry sieving) and relative methods (trapping, Direct observation during night). South concluded that the
water sieving is the most reliable method, since it allows the recovery of almost the 100% of the land snails
contained in one soil samples. Hunter (1968a) proved the efficiency of this method when he recovered almost all
individuals of a known population; South (1964) concludes that the water sieving is the most accurate method;
It´s also is the only reliable method to obtain and quantify land snail lays.
Almost all the authors agree that land snails and it´s lays are located in the uppermost stratum of the soil.
According to South (1964), 100% of the lays and land snails of D. reticulatum species is located in the first 2 or
11

3cm of soil in field areas. In crop areas, Hunter (1966) found out that 83% of individuals of D. reticulatum were
located in the soil first 7´5cm, and a 6% were located above 15cm. Also in crop areas, Runham & Hunter (1970)
observed that in the first 10cm of soil contained the 97% of D. reticulatum. Rollo & Ellis (1974) pointed out that
the 90% of snail lays are also located in the same soil layers. According to Marquet (1985), in standard conditions
any land snail appears above the first 5cm of soil, However in exceptionally severe winters up to a 15% of
individuals may appear at depths between 10 and 20cm. South (1992) suggests that sampling the soil´s first 10cm
it´s enough to quantify land snail populations.
Importancia: el WP. 1 es importante ya que nos va a proporcionar información sobre el tamaño,
estructura y dinámica de la población de caracoles y babosas plaga, información que emplearemos en el
desarrollo del modelo predictivo
Assessment of progress and results. Por medio de los informes semestrales y anuales, y por medio de los
controles personales que periódicamente el coordinador realiza a cada uno de los partner en el país
correspondiente.
--------------------------------------------------------------------------------------------------------
Task 1.1
Objetives
Studying the land snail population size in crops where they will conduct the study.
Participants: all partners
Materials and Methods
The methodology used is based on absolute estimation methods; land snails quantification of known surface soil
samples.
Soil sampling.
The choice of the sampling location is randomly selected. To carry this out the plot is divided in a grid composed
at least of 50 frames of 4 x 4 m; each one subdivided in four quadrants. The quadrants are determined using the
last two digits of the randomly generated value of the “random” (ran) calculator function. Once selected 20
frames, we proceed to determine which quadrant of each frame, by using again the calculator random number
generator following the next code: 0,000-0,249 for the upper left quadrant; 0,250-0,499 for the upper right
quadrant; 0,500-0,749 for the lower left quadrant; 0,750-0,999 for the lower right quadrant.
The sample extraction is performed with a rectangular spade mark with the depth to be achieved (10
centimeters). First is selected the area to take the soil sample, then we place on the ground an aluminum frame
of 25 x 25 cm, then the spade is stuck in to the ground along its entire contour until the marked depth and extract
the sample. Each sample is introduced in a properly labeled opaque plastic bag; these bags are conserved in a
cold storage at 4ºC in the dark, for its further laboratory analysis in the next 3 days.
To determine the number of soil samples needed its used an statistical software; The means and variances of the
average land snails and eggs are calculated, to find out all the possible combinations of 2 samples, 3samples,
4samples till the total of 20 samples. For each number of samples the degree of error is calculated dividing the
standard deviation by the arithmetic mean. In previous investigations we observed that to obtain a degree of
error less or equal to 10%, 18 samples are enough in the case of land snails and 16 in the case of eggs.
The abundance of land snails and eggs is estimated by washing the soil samples precedent from the study plots.
On a monthly basis 20 soil samples of 25x25 cm square and 10 cm deep.
Soil samples treatment. Each sample is placed individually on a white plastic tray; in the first place the sample is
thoroughly inspected to capture any snail that might be in the soil surface, once done this the vegetal cover is
removed, cutting it with scissors; then the soil sample is washed in sieves, with a decreasing mesh sizes from
4mm to 1mm. The soil samples are crumbled to smaller pieces with the help of the water jet. The thicker roots
are cut to smaller pieces. The sieves content is carefully inspected under a 10X magnifying glass and a powerful
white light source.
12

Larger land snails are retained in the upper sieve, and in the lower sieve the smaller land snails and the eggs. The
collected land snails are kept in a tray and the eggs in a Petri dish, both with a humid filter paper. Once finished
separating eggs and land snails from the soil samples we proceed to identify them.
---------------------------------------------------------------------------------------------------------------------
Task 1.2
Objetives
Field experiments to investigate
To know the land snails pests populations dynamic and structure.
Participant: All partners
Materials and Methods
To study the population structures and variation over time it´s necessary to determine the maturity state
of the land snails constituting the population along time. Because of this, Bett (1960), Hunter (1968a), and Hunter
& Symonds (1971), based their studies exclusively on sperms presence or absence along the genital tract. South
(1989a) used the same methodology, but he also obtained the gonad and albumin gland mass, (Hermaphrodite
Gland Index, H.G.I. and Albumin Gland Index A.G.I. respectively, of each individual. This index express the % of
corporal mass represented by each gland, that have a characteristically variation along the maturation cycle of D.
reticulatum.
Duval & Banville (1989) and Barker (1991), in adition to calculating the H.G.I. and A.G.I., incorporated in
their work the gonad cytological analysis of individuals, and determined their maturity level using as reference the
previous studies of Runham & Laryea (1968), based on the presence and relative abundance of each cellular type
in gonad gametogenisis in D. reticulatum gonad. Haynes et al. (1996) classified the land snails in five categories
based on the body mass index H.G.I. A.G.I. instead of using the cytological gonad analysis to describe the
population structure. The population dynamic and structure study are only done on the species that really are a
pest; in our previous studies (Barrada, 2003) we focused exclusively on Deroceras reticulatum that really
constitute a pest in the European crops.
Individual management. Individuals belonging to the pest species are weight on a laboratory balance to the
hundredth of milligram, and then they are sacrificed by a brief immersion in water at 50ºC. This method is a
modification from the Haynes, Rushton y Port (1996) method, which is to dip them in boiling water. The sacrificed
individuals are introduced in properly labeled glass tubes, with preservation 70% alcohol. Then the individuals are
dissected and they have their hermaphrodite gland and albumin gland removed, which is used to determine the
maturity state of individuals.
Following Barker (1991) approach, individuals with BMI(body mass index) exceeding 20 mg are not dissected,
assuming that they don´t have a differentiated gonad. The hermaphrodite and albumin gland, extracted from
individuals with BMI greater or equal to 20 mg, the glands are weighed up to the hundredth of milligram
immediately after its extraction. The hermaphrodite gland is fixed with Carnoy for 24 hours and preserved in
alcohol 70%.
Determining the maturity degree. It will follow the methodology used by Duval & Banville (1989) and Barker
(1991), the sexual maturity status of individuals at each monthly sampling is determined by gonad cytological
analysis, using as a reference the states defined by Runham & Laryea (1968) . To this end, each individual gonad
was dehydrated by a series of ethanol baths of progressively higher degree (70%, 96% and 100%), ending with 2
baths of toluene. Next the sample is included in a paraffin block; which is sectioned at 8 μ thick. Obtained sections
are rehydrated by reversing the previous dehydration, replacing toluene by xylene and dipping them in distilled
water. Then the sections are stained using hematoxylin-eosin staining, and finally dehydratated again.
Each cellular type that appear in the selected land snails gonad and the maturity states are available in Pelluet &
Watts (1951), Watts (1952), Bridgeford & Pelluet
(1952), Henderson & Pelluet (1960), Smith (1966), Runham & Laryea, (1968), Bailey (1973), Hill & Bowen (1976),
Parivar (1978, 1980, 1981), Nicholas (1984), South (1992), Fawcett (1987) and Lutchel et al. (1997) works.
13

Each captured individual was identified as belonging to one of the following sexual maturity stages: i)
undifferenciated spermatogonia, ii) spermatocyte, iii) spermatid, iv) espermatozoa, v) oocyte and vi) senescent.
The first three states correspond to immature land snails, with no reproduction ability; sexually mature land snails
are those that are in spermatozoon and oocyte state (Runham y Laryea, 1968; South 1989a).
In other words, each captured individual is characterized by their body mass (mg), by their maturity state
and by the mass (mg) of the hermaphrodite and albumin gland. From these values it´s calculated for each
individual, the hermaphrodite gland index (H.G.I.) and the albumin gland index (A.G.I.), as follows,
H.G.I. = Hermaphrodite gland mass 100 / individual mass
A.G.I. = Albumin gland mass 100 / individual mass
For each sample occasion, individuals who have similar characteristics referring to its sexual maturity
state and body mass index values, H.G.I. and A.G.I., are considered as belonging to the same land snail
generation.
In this research project we follow the methodology used by Duval & Banville (1989) and Barker (1991) for
the population structure study. As these authors did, we assume that individuals with body mass exceeding 20mg
are land snails with completely undifferentiated gonads (from the cytological viewpoint) are not analyzed. In this
regard, it should be mentioned that South (1989a) indicates a value of 40 mg body mass as a limit from which, the
maturity state of D. reticulatum can be defined by studying the cytology of the testis. Previous obtained results
agree with the values set by South (1989a).
14

Work Package 2
Field research to investigate the feeding habits and the qualitative and quantitative diet of land snails in
horticultural crops.
Objetives
To Study land snail feeding and qualitative and quantitative composition of their diet in order to find plant
species that can be used as trap plants in organic farming.
Partners 1 (……. Man-month)
Partners 2 (……. Man-month)
Partners 3 (……. Man-month)
Partners 4 (……. Man-month)
Partners 5 (……. Man-month)
Partners 6 (……. Man-month)
Partners 7 (……. Man-month)
Partners 8 (……. Man-month)
Partners 9 (……. Man-month)
Background
El estudio del material ya ingerido por los caracoles y babosas se realiza mediante el análisis microscópico
de los pequeños fragmentos de plantas que se encuentran en las heces o en el interior del tubo digestivo de los
animales. Este método ha sido utilizada por Grime, Blythe y Thornton (1970), Mason (1970), Wolda et al. (1971),
Chatfield (1975), Richardson (1975), Williamson y Cameron (1976), Szlavecz (1986), Speiser y Rowell-Rahier
(1991), Hatziioannou et al. (1994), para estudiar la dieta de caracoles como Cepaea nemoralis, Oxychilus cellarius
(Müller, 1774), Oxychilus alliarius (Miller, 1822), Discus rotundatus (Müller 1774), Arianta arbustorum (Linneo,
1758), Monadenia hillebrandi (Smith, 1957), Monacha cantiana (Montagu, 1803), Monacha cartusiana (Müller,
1774), Braybaena fructicum (Müller, 1774), Helix lucorum (Linneo, 1758), Xeropicta arenosa (Ziegler, 1827) y
Cepaea vindobonensis (Férussac, 1821). Hunter (1968b), Pallant (1969, 1972), y Jennings y Barkham (1975) la han
utilizado para estudiar la dieta de babosas como Deroceras reticulatum, Tandonia budapestensis (Hazay, 1881),
Arion hortensis Férussac, 1819, y Arion ater.
El estudio de las heces es un método más rápido que el análisis del contenido estomacal, ya que no
requiere el sacrificio y disección de los animales. Sin embargo, los materiales presentes en las heces han
atravesado todo el tubo digestivo del animal y se encuentran más degradados, por lo que resultan más difíciles de
identificar que los extraídos del estómago (Cook y Radford, 1988; Hatziioannou et al., 1994). El estudio de heces
es un método adecuado para determinar la presencia o ausencia de determinados elementos en la dieta de los
animales, pero la proporción de materiales no identificados en las heces suele ser tan elevada, que no resulta un
método útil para realizar una caracterización cuantitativa de la alimentación (Williamson y Cameron, 1976;
Szlavecz, 1986; Speiser y Rowell-Rahier 1991). Vadas (1977) indica que los materiales más abundantes y más
fácilmente identificables en las heces de los animales son los menos utilizados metabólicamente y que,
posiblemente, son también los ingeridos en menor cantidad, por lo que el análisis de heces conduce a una
sobrevaloración de los alimentos poco consumidos; por el contrario, los alimentos más consumidos son
infravalorados debido a que son digeridos en mayor medida y resultan más escasos y difíciles de identificar en las
heces.
El análisis del contenido del tubo digestivo de los animales es un método más laborioso, pero proporciona
una imagen más real de la dieta de los animales (Norbury y Sanson, 1992). Pallant (1969, 1972) estudió la
alimentación de D. reticulatum en poblaciones naturales (bosques y praderas) mediante el análisis de contenidos
estomacales. Para reducir al máximo la degradación digestiva del alimento ingerido por los caracoles y babosas
capturadas y facilitar su identificación, Pallant (1969, 1972) introdujo directamente a los animales capturados en
alcohol al 70%, y realizó su disección para la extracción del contenido del tubo digestivo a lo largo de las 2 horas
siguientes a la captura. Triebskorn y Florschutz (1993) realizaron un estudio sobre el tránsito del alimento a través
del tubo digestivo de D. reticulatum, mediante la utilización de un preparado alimenticio (lechuga, maíz y leche en
polvo) marcado radiactivamente y la toma de radiografías de los animales a intervalos regulares; según sus
resultados, el alimento ingerido penetra inmediatamente en el buche, estómago y parte anterior del intestino,
lugares en los que permanece durante un período mínimo de dos horas y media antes de comenzar a avanzar por
15

el intestino; el buche y el estómago se vacían completamente en el curso de las trece horas siguientes a la
ingestión.
Importancia: la información que se obtenga de este estudio nos servirá para diseñar las estrategia del uso
de plantas-trampas como agentes disuasorios y conocer y evaluar los daños reales sobre los cultivos.
Assessment of progress and results. Por medio de los informes semestrales y anuales, y por medio de los
controles personales que periódicamente el coordinador realiza a cada uno de los partner en el país
correspondiente.
Work and methodology Description
Sampling. The diet study is done by the analysis of the digestive tract content of 20 land snails captured in
monthly samplings. The captures of these animals are randomly done in the course of a run across the whole plot.
Only the collection of small land snails is avoided, because their processing is very difficult. The capture of the
land snails is done during the next 3 or 4 hours after sunset, which is when it takes place the main feeding activity
of land snails and D. reticulatum in particular (Dobson & Bailey, 1982, Rollo, 1988ab; South, 1992; Hommay,
Lorvelec & Jacky, 1998). Every plant species, where each land snail is captured, is recorded. The land snails are
individually introduced in plastic containers with a piece of damp cotton for its further transport and storage in
the laboratory.
The day after the capture of the land snails, the plot´s vegetation composition is determined. The
different plant species coverage percentage is estimated using a measuring tape extended along the plot´s
principal axis, at intervals of 0.5 meters. The tape´s intersection points with the different plant species are noted,
and with this data the coverage percentage for each species is calculated. Graminaceous are considered as a
whole.
Individual Processing. The land snails caught for the diet analysis are transported and processed within 2
hours, in order to minimize digestive degradation of their stomach contents (Pallant, 1969, 1972).
Land snails are weighed on a scale up to the hundredth of milligram, before being sacrificed by immersion
in hot water (50 º C). Then the individuals are dissected and have their maw removed, which is placed onto a
slide. Under a binocular microscope the maw is open along the longitudinal axis and all its contents are collected,
using a fine brush. Each land snail´s stomach contents is weighed to the hundredth of milligram and immediately
introduced into a small plastic tube, were 2 milliliters of 1N hydrochloric acid is added to eliminate the mucus and
epithelial debris from the maw sample (Hatziioannou et al., 1994). Stomach contents are always analyzed in the
following two days after its capture.
Qualitative diet determination. The qualitative diet study is based on the food fragments identification,
found in the snails´ digestive tract. The plant fragments identification is possible through epithelial formations
such as stomata and trachoma. The appearance and distribution of these formations are characteristic for each
plant species.
Before starting the samplings, land snails are captured around the study area, they are fed with a
monospecifical plant diet, and their feces are used to make an image collection of the fragments found in the land
snails´ stomachs. These land snails are kept inside a climate chamber with a photoperiod of 12 hours, 18 º C and
90% relative humidity. All land snails are housed inside transparent plastic boxes 20 × 20 × 10 cm, with the
bottom covered with a damp filter paper, and all the plastic boxes are perforated to allow the air renewal. In each
case four individuals are kept. After a period of 72 hours without food all land snails will be fed with a diet
consisting on a single plant species collected in the study plot. Then after six hours, the land snails are processed
as described in the previous section, and the fragments found in their stomachs are photographed under a
microscope (Barrada, 2003).
For each plant we have a large reference image collection of each one after being eaten by land snails.
The captured land snail´s digestive tract is studied using the same microscope equipment. Fragments found in the
land snails´ digestive tract are compared with the reference images collection for their further identification.
Quantitative diet determination. To determine the land snails´ quantitative diet composition, we follow
the methodology used by Hatziioannou et al. (1994). A stomach sample content of each land snail is taken; the
surface area of each fragment is measured with the help of a software image analysis (SPSS ® Sigma Scan Pro
Image Analysis Version 5.0.0.). The surface areas of all the same type fragments are added together and the
percentage is calculated and represented in relation with the sum of all the fragments contained in the same
sample surface areas. According to this, each plant contribution to the land snail diet is estimated.
16

Using 0.24 ml sample taken from the stomach contents of each land snail, diluted with 2 ml of
hydrochloric acid and then homogenized with the aid of an agitator pressure SBS ® AT-1 or similar. All fragments
of food contained in the sample are photographed, identified and measured.
The sample volume used for each digestive tract is previously determined from the land snails´ stomach
contents study, whose diet is known. To this purpose four land snails are individually housed in each plastic box.
After a fasting period of 72 hours, the land boxed land snails are provide with a small piece of 3 different plant
species, and 6 hours after they are sacrificed and their stomach contents are removed, as previously described.
Their stomach contents are diluted in 2 ml of HCl for their analysis in 6 successive 0.08 ml samples. In previous
researches (Barrada, 2003), it was determined that a 0.16 ml sample (8% of total volume) is an accurate
representation of the land snails stomach contents composition, equivalent to a 0.48 ml sample (24% of the total
volume). To minimize the error degree it is always used a 0.24 ml sample (12% of total volume) for the digestive
tract contents representation.
Diversity and selection Indexes. The diversity land snails´ diet and the vegetation variety in the study
plots, is calculated through the Shannon-Weaver index, H '= Σ (pi) (log2pi), where pi is the frequency for each
component of the diet or vegetation (Margalef, 1982).
The index C is used as a selection index, Pearre (1982), this index reflects the outcome of predator-prey
interaction taking into account the abundance in the environment for each prey type. This index has a value
ranging from +1 and -1, where C = 0 indicates no selection. It shows if a plant is consumed above (positive values)
or below (negative values) it´s expected consumption according to their availability in the nature. This index is
based on the χ 2 test that allows establishing the significance of the selection degree for any sample size.
17

Work Package 3
Objetives
To develop a statistical method to explain and predict the land snails´ activity according to atmospheric
conditions.
Partners 1 (……. Man-month)
Partners 2 (……. Man-month)
Partners 3 (……. Man-month)
Partners 4 (……. Man-month)
Partners 5 (……. Man-month)
Partners 6 (……. Man-month)
Partners 7 (……. Man-month)
Partners 8 (……. Man-month)
Partners 9 (……. Man-month)
Background
La actividad de los gasterópodos terrestres está regulada por complejos mecanismos en los que
intervienen tanto factores externos (ambientales) como internos (ritmos endógenos) (Bailey y Lazaridou-
Dimitriadou, 1986; Aupinel, 1987; Young y Port, 1989; Cook, 2001).
Como regla general, se admite que estos animales pasan el día inactivos en sus refugios, que su actividad
es principalmente nocturna, y que las condiciones meteorológicas determinan en gran medida el que se muestren
más o menos activos (Hommay et al., 1998). No obstante, se reconoce la existencia de diferencias interespecíficas
en lo que se refiere a sus patrones de actividad y a la influencia que ejercen distintos factores ambientales sobre
los mismos (Cook, 2001). También se ha apuntado la existencia de diferencias intraespecíficas en la importancia
relativa de los distintos factores ambientales que controlan la actividad, en función del microclima o del tipo de
ambiente en el que viva cada población (Lorvelec y Daguzan, 1990; Iglesias y Castillejo, 1996). La estrecha
dependencia que presentan los gasterópodos terrestres con respecto de las condiciones ambientales hace de
ellos unos modelos ideales para el estudio de la relación existente entre el comportamiento animal y el clima
(Rollo, 1982). Su carácter de plagas agrícolas es otro aspecto que contribuye a explicar el interés demostrado por
numerosos investigadores en el estudio de su actividad en relación con las condiciones climáticas (Dainton,
1954ab; Getz, 1963; Webley, 1964; Newell, 1968; Cook y Ford, 1989; Young y Port, 1991; Young, Port, Emmet y
Green, 1991; Hommay, Lorvelec y Jacky, 1998; Grimm y Kaiser, 2000, Barrada, 2003).
Una de las formas de abordar ese estudio, que está recibiendo una gran atención en los últimos años
debido a su aplicabilidad en el campo del control de plagas, es la elaboración de modelos de predicción o modelos
expertos (Bohan et al., 1997, Cook, 2001). Los modelos de predicción de actividad de animales con carácter de
plaga representan una herramienta muy útil desde el punto de vista aplicado, puesto que permiten pronosticar
períodos en los que los cultivos pueden sufrir un mayor daño y optimizar la utilización de los plaguicidas
empleados para su control, reduciendo los costes económicos y ambientales que se derivan de su aplicación en
momentos en los que no existe riesgo de daño para los cultivos (Frahm, Johnen, y Volk, T. 1996; Hommay et al.,
1998; Hommay, 2002; Port y Ester, 2002).
Importancia: los datos obtenidos después de esta investigación nos servirán para desarrollar un modelo
estadístico de predicción abundancia y actividad.
Assessment of progress and results. Por medio de los informes semestrales y anuales, y por medio de los
controles personales que periódicamente el coordinador realiza a cada uno de los partner en el país
correspondiente.
Methodology
The Barrada(2003)methodology is followed to develop the activity models, this methodology is based on
the ones designed by Young & Port (1989) and Young et al. (1991). These authors identified, for the diverse
environmental conditions, the limits that define the high land snail activity nights, without attempting to
mathematically describe the limits that define those lines, but only giving the extreme values to define the
18

optimal range values for each variable. These models predict that high activity nights will be those with all the
variables comprehend between their optimal ranges.
Sampling. The land snail´s activity analysis is done in the same study area used to analyze its feeding
habits. The samplings are done during three consecutive nights per month; this means a total of 72 nights during
the sampling period. Every night two investigators examine during three hours the study area, searching for active
land snails. The tours are done thoroughly, searching the soil and vegetation, but no effort is done to locate those
out of the observer´s sight.
At the beginning of each sampling and subsequently, at 1 hour intervals, the soil temperature and the
relative humidity are registered at 5 cm above ground. The soil temperature can be registered with an electronic
thermometer fixed in the ground at 10 cm deep, and the temperature and the relative humidity are registered
with an electronic thermo-higrometer or a similar instrument.
Variables and Statistical analysis. In the model, the activity is divided in three categories (low, average
and high) according to the number of active land snails registered during 72 nights in each study area. This activity
levels made up the dependent models or response variables, in other words, (the variable whose value was
expressed in terms of the values of the independent or predicted variables). Within these environmental
variables were considered measures on site during the samplings are considered as independent variables
measures on site during the samplings is considered as independent variables: i) the atmospheric conditions
registered on site during the samplings; ii) atmospheric conditions corresponding to the sampling day and the
days before it, according to the data registered in a near thermo-pluviometric station; iii) time variables (month,
season); and iv) related variables with the land snails population dynamics in the sample area. All considered
variables are compiled in table 5.1.
Because of the data nature, qualitative response variables with more than two categories (three
categories: low, average and high) and a serial of qualitative (factors) and quantitative (co variables) variables as
independent variables, the statistical procedure to relate the land snails activity level with the independent
variables was the ordinal regression (McCullagh, 1980; McCullagh & Nelder, 1989). To carry out the data analysis
it can be used the SPSS, using the PLUM method for the ordinal regression (SPSS documents), or any related
statistic programs (Barrada, 2003).
19

Work Package 4
Searching for a Biopesticide from plant extracts with molluscicide and ovicide activity.
Partners 1 (……. Man-month)
Partners 2 (……. Man-month)
Partners 3 (……. Man-month)
Partners 4 (……. Man-month)
Partners 5 (……. Man-month)
Partners 6 (……. Man-month)
Partners 7 (……. Man-month)
Partners 8 (……. Man-month)
Partners 9 (……. Man-month)
Objetives
To investigate the feasibility of using plant extracts as biomolluscicides and bio-ovicides.
Background
Farmers in their traditional wisdom have identified and used a variety of plant products and extracts for pest
control, especially in storage. As many as 2121 plant species are reported to possess pest management
properties, 1005 species of plants exhibiting insecticide properties, 384 with antifeedant properties, 297 with
repellant properties, 27 with attractant properties and 31 with growth inhabiting properties have been identified.
The most commonly used plants are neem (Azadirachta indica), pongamia (Pongamia glabra) and mahua
(madhuca indica). 2-5 % neem or mahua seed kernel extract has been found effective against rice cutworm,
tobacco caterpillar, rice green leafhopper, and several species of aphids and mites. The efficacy of vegetable oils
in preventing infestation of stored product pests such as bruchids, rice and maize weevils has been well
documented. Root extracts of Tagetes or Asparagus as nematicide and Chenopodium and Bougainvillea as
antivirus have also been reported 'Biopesticide' can reduce pesticide risks, as- (a) Biopesticides are best
alternatives to conventional pesticides and usually inherently less toxic than conventional pesticides. (b)
Biopesticides generally affect only the target pest and closely related organisms, in contract to broad spectrum,
conventional pesticides that may affect organisms as rent as birds, insects, and mammals. (c) Biopesticides often
are effective in very small quantities and often decompose quickly, thereby resulting in lower exposures and
largely avoiding the pollution problems caused by conventional pesticides. (d) When used as a fundamental
component of Integrated Pest Management (IPM) programs, biopesticides can greatly decrease the use of
conventional pesticides, while crop yields remain high. (e) Amenable to small-scale, local production in developing
countries and products available in small, niche markets that are typically unaddressed by large agrochemical
companies.
The European Commission has published a proposed European Regulation concerning the placing on the market
and use of biocidal products. The new European Regulation would replace the current regulatory regime for
biocides, which is laid out in the Biocidal Products Directive 98/8/EC and transposed into UK law by the Biocidal
Products Regulations 2001. Once in force (scheduled for January 2013), the European Regulation would be
directly acting on all Member States, including the UK.
Methodology
1.-MATERIAL AND METHODS
Preparation of the extracts for the tests:
20

The selection of the plants will be made using two parameters: abundance and medicinal and therapeutic
properties. Previously it is necessary to consult references on bibliography.The plants will be collected in the field
and identified.
The extracts will be made with different parts of the plant, so much fresh as after drying, in an oven at 50ºC
(Medina and Woodbury, 1979 & Makkar et al 1991) or at room temperature. The different parts of the plant
(leaves, steams, fruits, flowers, roots and seeds) will be separated and crushed to powder. The powder of the
plant will be put in glass flasks, where will be spilled the solvent, leaving it macerates during 24 hours at room
temperature. Passed these 24 hours the samples with acetone/water will be extracted with the help of a Rotary
Evaporator machine (Mendes et al, 1993).
The concentrations used will be: 80,000 ppm, 40,000 ppm and 8,000 ppm to the water extracts; 100,000 ppm
and 50,000 ppm to the extracts made with acetone/water; and: 100,000 ppm, 50,000 ppm, 10,000 ppm, 1,000
ppm and 100 ppm for both extracts.
Mollusciciding activity:
The tests will be made using 10 cm diameter glass Petri dishes with a 9 cm diameter filter paper on the bottom
(Albet 400, of 90g/m 2 of weight and a thickness of 0.21mm). 1 ml of extract will be put in each dish, using a
pipette that had a milk filter (Alfa Laval Agri) in the tip with the purpose of avoiding that the pipette was plugged
with the powder of the plant. The dish with the filter will be driest off at room temperature; once it was
completely dry it will be moistened with 1 ml of distilled water, then the eggs will be deposited on the filter (5
eggs in each dish, previously the eggs had been selected). They stayed in the incubation camera until the death or
hatching. To check the effect of the extract on the embryo the eggs were observed, each 24 hours, with the
binocular magnifying glass using a glass tube (oviscope).
The controls of the water extract will be made with distilled water, while controls for the acetone/water
extracts were made with an acetone/water mixture (in the proportion: 7:3), then the acetone will be removed
with the Rotary Evaporator and the water was used to the controls.
To test it 1 ml of the water will be put on the filter papers, when the paper dry off at room temperature
another millilitre was put on it and then the eggs were placed on the moist paper. The Petri Dishes were closed
and were put in the incubation camera.
The same process will be made with the standard soil and with commercial substratum. At this time
plastic Petri dishes of 9 cm of diameter were used to these purposes. 1 ml of extract was put on 25g of standard
soil with a humidity of 35%, and the same process was made with the substratum.
Task 4.1.
To carry out laboratory experiments to find plant extracts to be used as biopesticida to control agricultural land
snail’s pest.
Objectives.
Laboratory tests on filter paper (direct contact) and standard soil to select plant extracts with molluscicidal and
/or ovicidal activity.
Materials and Methods
Egg-lays. The land snails are collected in the crops to obtain the egg-lays to make the experiments once in
the laboratory. The individuals are kept in plastic boxes (25 x 25 x 15 cm) with perforated walls and lids and the
floor covered with a moist filter paper. Black small polyethylene tube pieces are used as shelters for land snails,
using as food, lettuce, carrots, cabbages, runner beans, potatoes, and mushrooms, supplemented with powdered
CO3Ca. The cages are placed in a climatic chamber at 17ºC day/15ºC night with a 12D:12L photoperiod and 85%
21

elative humidity. Cleaning and food replacement will be done twice weekly. Since land snail breeding takes place
whenever environmental conditions are suitable (Carrick, 1938; South, 1989), the cages are inspected looking for
egg-lays every day. The land snails laid their eggs directly on the filter paper, mainly in those places covered by
pieces of food. The eggs will be collected, cleaned with distilled water and incubated on wet filter paper inside
Petri dishes in the darkness at 18ºC. The entire course of the development of the embryo is observable when the
egg is immersed in water and viewed under transmitted light (Carrick, 1938). About a week after collection, all the
eggs will be inspected under a binocular microscope for the selection of those to be used in the experiments. Only
eggs containing a single living embryo and without foreign inclusions will be selected for the tests, and they will
be used when the embryo achieved the developmental stage IV according to Carrick (1938), which is recognisable
by the differentiation of rudiments of the tentacles and posterior sac. The movement of the embryo in the form
of a slow and continuous rotation, and the rhythmical contractions and expansions of the still-small posterior sac
were the criteria used to determine whether the embryo was alive. Non-motile embryos were considered to be
dead. Carrick (1938) described the structure of the egg and the embryonic development of Deroceras reticulatum,
while the histochemistry of the egg was described by Bayne (1966, 1968) and Barrada (2003).
Contact toxicity tests on artificial soil. The tests will be made in glass Petri dishes 9 cm in diameter,
containing 40 g of standard artificial soil. The standard soil (OECD, 1998) consisted of 10% peat, 20% kaolin and
70% quartz sand. The pH will be adjusted to six with calcium carbonate and soil moisture was set to 35% (w/w).
All tests consisted of a control and five doses of the compound, arranged in a two-fold geometric series, with five
replicas each and five eggs per replica. For each compound, the appropriate quantity will be dissolved or
suspended in 20 ml of distilled water. The appropriate quantity of compound will be calculated, according to its
formulation, to render the desired maximum concentration of a.i. cm -2 when applying two ml of the solution or
suspension to the soil surface of one dish. Thereafter, ten millilitres of that solution or suspension will be applied
to the soil (2 ml per replica of the highest concentration) and the remain ten millilitres will be diluted to 50% to
obtain the next lower concentration. Dilutions and application of the solutions or suspensions will be made under
continuous shaking. The different treatments will be applied spraying 2 ml of the solution or suspension on each
Petri dish. Two ml of distilled water will be applied to the control dishes. After treatment, the soil in the Petri
dishes will be allowed to dry for 24 hours at room temperature. Immediately before the start of the test, the soil
was moistened again with distilled water in an amount calculated from the weight loss of the controls. Then, five
eggs will be placed on different points of the soil surface of each dish. After the start of the experiments, all the
eggs will be inspected under a binocular microscope every 24 hours to assess whether the embryos were still
alive. To avoid immersion of the eggs in water to view them under the microscope, which would produce an
undesirable wetting of the eggs, the test-eggs will be introduced into a glass tube, 3 mm inner diameter; in this
way through the contact zone between the egg and the glass is possible to see the embryo inside the egg
producing the same effect as immersion in water. Median lethal doses (LD50), i.e. the calculated doses which
produce 50% mortality of the eggs, and 95% confidence limits, will be calculated by probity analysis. For the
different compounds, LD50s will be calculated for periods of exposure in which at least one of the doses tested
resulted in no mortality and one in 100% mortality (OECD, 1998, Guideline 313 de la OECD (OECD Guidelines for
the Testing of Chemicals. Contact toxicity tests on artificial soil/ wet filter paper). (Iglesias, J., Castillejo, J., Parana,
R., Mascato, P. and Lombardía, M.J. 2000; Iglesias, J., Castillejo, J. y Ester, A., 2002; Iglesias, J., Castillejo, J., Ester,
A. y Lombardia, M.J., 2002)
Task 4.2
Testing biomollusicicede activity of plant extracts on agricultural soil mini plots.
Objetives.
Mini plots experiments on horticultural soil to evaluate the efficacy of the selected plant extracts against lands
snails and its eggs
Materials and Methods
TEST PLOTS
In each study plot six areas are defined in squares of 4 x 4 m= six Mini Plots of 16 m 2 . Plant extracts with a
specific composition are tested in five mini plots and the sixth acts as the control plot. Between each mini plot
there is a security area of 2 m wide.
22

In each mini plot are placed five Bayer trap-shelters specific for land snails and five PVC tubes specific for
snails. These tubes are our invention; they are PVC tubes of 30cm long and 5 cm in diameter and closed at one
end. They are placed on wooden supports fastened and nailed around the crop plants, with its hole facing down
so that the snails take shelter inside (Córdoba, 2009).
PLANT EXTRACTS CONCENTRATION AND APPLICATION METHODS
From each plant extract with ovicidal action we know the LD50 in grams per square meter, we
also know the surface that will be treated (4 x 4 m 2 ), and we know how much depth we can find in the soil the
eggs of land snail, so it will be easy to calculate the amount to apply in each mini-plot (to be applied in every mini
plot.).
Knowing when to apply the plant extract one with ovicidal action, that is to know the period of the year
to be applied (in which it is necessary to apply that) is determined by the life cycle of land snails, pests that have
been studied in the first year of the project, that is to say it will be applied when the population density of land
snails, land snails and eggs in the soil is minimum. So many applications will be done of agrochemical as valleys or
sinuses that have the life-cycle curve of the land snail in the study area.
Shelter traps are placed after each one of the treatments and are removed during the application of
agrichemicals (the agrochemical one.).
SAMPLING
The time length: 24 consecutive months (2 years). Sampling frequency: monthly. Before carrying out the
first application of the agrochemical one the density of the towns will be estimated of land gastropods in the
pieces of land object of study will be estimated following the exposed methodology in Work Package 1, Task 1.1.
After application of plant extracts with ovicidal action, with monthly regularity and for 2 consecutive
years, it will be estimated in each area, the land snails population density, land snails and eggs of these found
under the shelter traps.
Every month they will lift the shelter traps and identify the land snails, all captured individuals will be
measured and weighted. The eggs found under the traps will be identified and counted (Cordoba, 2009).
Every six months a statistical analysis will be done on data obtained with the population density of land
snails and egg-lays. After the first year of testing, and after the harvest collection, (once collected the harvest,) or
once the crop, will make another estimate of population densities of land snails and egg-lays in the soil according
to the methodology outlined in Work Package 1, Task 1.1.
CROP DAMAGES EVALUATION
After plant extracts treatment, land snail damages will be assessed after 3 days and thereafter at monthly
intervals during crop life. Damage will be recorded as the percentage of leaf area eaten (percentage leaf loss) to
the nearest 5% and will be assessed separately for each plant. For statistical analyses the recordings of 20 plants
growing in the same plot were averaged, because they reflect the activity of the same land snail population,
resulting in six independent assessments for every treatment. A random program will be used for 20-plants
selection. Data for land snail damage (percentage leaf loss) will be transformed to angles prior to analysis of
variance. Data for number of damaged plants and for number of land snails were compared non-parametrically by
the Mann-Whitney U-test. (Iglesias, Castillejo and Castro, 2001, 2001b and 2003)
Task 4.3 Mini plots analysis to know the collateral effect of plant extract selects on invertebrate soil.
Materials and Methods
In areas near the test plots, and following the methodology of sampling and analysis presented in the
Work Packcage 1, Task 1.1, soil samples are taken to study side effects. On a sieve of appropriate mesh size
worms, mites and springtails are collected. To determine the possible toxicity of the chemicals used in the study
plots tests will be conducted recommended by the OECD GUIDELINES FOR TESTING CHEMICALS (1th draft,
November, 2007 Proposal for a New Guideline) on earthworms, mites and springtails.
23

The assessment (valuation) of edge effects on terrestrial gastropods in wild areas near and without plant
extracts treatment (untreated) it will be applied the same methodology of the Bayer type trap shelters and PVC
pipes has now been exposed (WP. 4, Task 4.2.)
Task 4.4 Chemical analysis to find the plant extracts active principle by analytic steps.
These kinds of analytic analysis and posterior test on eggs, snails and slugs will be carried out by Syngenta
Company at Switzerland laboratory in collaboration with Partner 1 (Coordinator) for effectiveness as molluscicide
and ovicide activity against slugs, snails and eggs and parallels studies on collateral effects on soil fauna. These
analyses and the USC collaboration will be out of global work packages of this project
24

Work Package 5
Destroy land snail egg-lays with non residual agrochemical compounds.
PARTICIPANTS
Partners 1 (……. Man-month)
Partners 2 (……. Man-month)
Partners 3 (……. Man-month)
Partners 4 (……. Man-month)
Partners 5 (……. Man-month)
Partners 6 (……. Man-month)
Partners 7 (……. Man-month)
Partners 8 (……. Man-month)
Partners 9 (……. Man-month)
OBJETIVES
To investigate the feasibility of using commercial agrochemical with ovicidal activity to kill control land snail eggs
for key conventionally grown horticultural crops
BACKGROUND
According to Mendis et al. (1996), among the many methods proposed for controlling gastropod pests in
agriculture, the eggs of the animals have received very little attention. Particularly, there are very few data on the
effect of pesticides or other substances on the viability of terrestrial gastropods eggs (Stringer & Morgan, 1969,
1970, 1972, cited by Godan, 1983; Ryder & Bowen, 1977a). The high susceptibility of the eggs of D. reticulatum to
contact with low doses of metal salts (Iglesias et al. 2000) and with some pesticides (Iglesias, Castillejo & Ester,
2002) has been demonstrated recently in paper contact-toxicity tests.
Importancia: Los molusquicidas tradicionales solo matan algunos caracoles y babosas, pero con el uso de
ovicidas podremos destruir todos los huevos del suelo, estos ovicidas son compuestos agroquímicos
habitualmente usados por el agricultor.
Assessment of progress and results. Por medio de los informes semestrales y anuales, y por medio de los
controles personales que periódicamente el coordinador realiza a cada uno de los partner en el país
correspondiente.
WORK AND METHODOLOGY DESCRIPTION
See tasks
-----------------------------------------------------------------------------------------------------
Task 5.1
To carry out laboratory experiments for the selected commercial non residual agrochemicals with ovicidal action.
Objectives
To make laboratory tests on filter paper (direct contact) and on standard soil to select the non residual
commercial agrochemicals that are more effective against land snails lays, for their posterior use as ovicides in
crops.
Agrochemical selection. Searching for non residual agrochemicals compounds with ovicidal activity.
Agrochemicals used in the tests are the ones approved by the local authorities.
See Work Package 4, Task 4.1. Only it is necessary to change plant extracts for non residual agrochemicals.
Task 5.2
Field experiments to investigate the feasibility of using non residual agrochemicals compounds with ovicidal
activity of killing land snail eggs.
25

Objetives
Field experiments on horticultural crops to evaluate the efficacy of selected agrochemical as molluscicide-ovicicde
of killing land snail eggs of control for key conventionally grown horticultural crops. Final trial.
Materials and Methods
See Work Package 4, Task 4.2. Only it is necessary to change plant extracts for non residual agrochemicals.
Task 5.3
To investigate the collateral effect of agrochemical compounds on soil fauna and wild land snails.
Objetives
Field analysis to know the collateral effect of agrochemical selects on invertebrate soil fauna and border effect on
wild land snails in conventionally horticultural crops.
Materials and Methods
See Work Package 4, Task 4.3. Only it is necessary to change plant extracts for non residual agrochemicals
26

Work Package 6
To carry out laboratory and field experiments to investigate the feasibility of using livestock manure and
trap-plants as land snail pest control in non human consumption organic farming.
PARTICIPANTS
Partners 1 (……. Man-month)
Partners 2 (……. Man-month)
Partners 3 (……. Man-month)
Partners 4 (……. Man-month)
Partners 5 (……. Man-month)
Partners 6 (……. Man-month)
Partners 7 (……. Man-month)
Partners 8 (……. Man-month)
Partners 9 (……. Man-month)
Objetives
To investigate the cattle and swine manure feasibility as a land snail eggs-lays killer and the use of trapplants
as a control strategy against land snail pests for key organic horticultural farms.
Background
El uso de purines animales como abonos orgánicos está muy extendido en la agricultura, el conocimiento
de su toxicidad sobre la fauna edáfica está demostrado. En ensayos previos el Partner 1 ha demostrado que los
purines de cerdo y vaca a una concentración baja tienen capacidad ovicida, concretamente hemos podido ver que
para eliminar todos los huevos de caracoles y babosas de una hectárea de cultivo hay que esparcir 10000 kg de
cattle slurrry o 8000 de swine slarry, la destrucción de los huevos se realiza antes los 5 días de la aplicación.
La utilización de cultivos-trampa es una de las alternativas que está recibiendo mayor atención por parte
de los investigadores: esta estrategia se basa en la coexistencia, junto a las especies cultivadas, de otras con
escaso o nulo valor económico que sirvan de alimento para los caracoles y babosas y que le resulten más
atractivas que las especies cultivadas. La gran mayoría de las investigaciones realizadas en este sentido son
experiencias de laboratorio en las que se buscan aquellas especies que resultan más atractivas para los caracoles
y babosas que las especies cultivadas, pero los resultados obtenidos en las mismas difícilmente se pueden
extrapolar a situaciones reales de campo, en donde la dieta de los caracoles y babosas está condicionada de
forma muy fuerte por la disponibilidad de las diferentes especies. Las especies utilizadas como cultivos-trampa
deben de cumplir su función protectora siendo poco abundantes, ya que compiten por los nutrientes con las
especies cultivadas. En el presente trabajo encontramos que D. reticulatum seleccionó la especie silvestre
Sonchus oleraceus para alimentarse. Esta especie realizó una contribución significativa a la dieta de los caracoles y
babosas pese a ser una planta poco abundante en la plota de estudio, lo que hace de ella una buena candidata
para ser utilizada como cultivo-trampa. Su capacidad para reducir los daños ocasionados por los caracoles las
babosas a las especies de cultivo debería ser evaluada en experimentos de campo.
Los purines de cerdo y vaca se usualmente son usados como abonos en prados y en cultivos extensivos, en
nuestro caso para los ensayos consideramos los purines como un compuesto agroquímico más, y la metodología a
emplear será la misma.
Importancia: El empleo cattle and swine manure como ovicidas para matar los huevos de los caracoles y
babosas es novedoso, y pone en mano de los agricultores ecológicos una herramienta muy útil.
Assessment of progress and results. Por medio de los informes semestrales y anuales, y por medio de los
controles personales que periódicamente el coordinador realiza a cada uno de los partner en el país
correspondiente.
WORK AND METHODOLOGY DESCRIPTION
Plot selection. To carry out this study, an organic farming plot is selected, were no chemical pesticides nor
fertilizers are used.
Cattle and Swine manure. The cattle and swine slurries are considered as animal origin agrochemicals suitable for
ecological farming.
----------------------------
27

Task 6.1.
To carry out laboratory tests with cattle and swine manure to determine their ovicidal action against land
snail egg-lays.
Objetives
To carry out laboratory tests to determine the effective concentration of the cattle and swine slurries
against land snails and egg-lays. The linear discriminant analysis done on filter paper (direct contact) and on
standard soil, first and second screening.
Material and Methods
The methodology set out in Work Package 4, Task 4.1 for agrochemical compounds will be followed.
-------------------------------
Task 6.2.
Objetives
Field experiments to evaluate the cattle and swine manure efficiency as land snail eggs-lays control for key
organic horticultural crops. Final trial.
Material and Methods
The methodology set out in Work Package 4, Task 4.2 will be followed.
Plant’s crops damage evaluation
See Work Package 4, Task 4.2.
-------------------------------
Task 6.3.
Objetives
To carry out field analysis to find out the cattle and swine manure collateral effects on invertebrate soil fauna and
the border effect on wild land snails in organic horticultural crops.
Material and Methods
The methodology set out in Work Package 4, Task 4.3 will be followed.
-------------------------------
Task 6.4.
Objetives
Carry out field experiments to use trap-plants as a deterrent method of protecting organic horticultural
farms.
Material and Methods
Trap-plants. One of the consequences of the development of WP.2 is to find out which plants that exist in the
study area is more attractive for land snails. These plants have a high selection index and remain low in
abundance. The trap-plants have little or no nutritional valor, these plants have to coexist with the crop plants
and at the same time be an attractive alternative food source for pests, to reduce the damages done to the
principal crop. This plants must also be in a low abundance, so that they don´t compete for the resources with the
principal crop.
Methodology. To design the mini plots tests the followed methodology is the same as the one explained in WP.5,
Task 4.2. The trap-plants are set around the mini-plots, as a stockade or in between the crop plants. The organic
farming damages will be done by estimating the foliage loss or estimating the number of damaged leaves, caused
by land snails, to evaluate the effectiveness of the trap-plants.
28

Work Package 7
Study the effectiveness of agrochemicals with ovicidal action on conventional horticultural crops.
PARTICIPANTS
Partners 1 (……. Man-month)
Partners 2 (……. Man-month)
Partners 3 (……. Man-month)
Partners 4 (……. Man-month)
Partners 5 (……. Man-month)
Partners 6 (……. Man-month)
Partners 7 (……. Man-month)
Partners 8 (……. Man-month)
Partners 9 (……. Man-month)
OBJETIVES
Field experiments in conventional key horticultural crops to evaluate the efficacy of selected agrochemical with
ovicidal action against land snail egg-lays in relation to other commercial standard low-chemical methods of
killing gastropods. Final trial.
BACKGROUND
Control integrado de plagas y modelos de predicción. Tradicionalmente, el concepto del control de plagas en
la agricultura planteaba, como objetivo básico, la eliminación total del agente causante de la plaga, mediante la
aplicación de pesticidas, cuando ésta era detectada en un cultivo. En los años 60, en Europa y Estados Unidos,
surge el concepto del control integrado de plagas (CIP) (Stern, Smith, van der Bosch y Hagen, 1959), que en la
actualidad es parte integrante de otro concepto, más amplio, que es el del desarrollo sostenible. El control
integrado de plagas implica la integración de los conocimientos provenientes de multitud de campos (biología,
química, agronomía, climatología, economía, etc.) con el fin de desarrollar las estrategias de control más
adecuadas desde el punto de vista económico, ambiental y de salud pública (Dent, 1991). Si bien es un sistema
basado en la combinación de diferentes métodos con el fin de minimizar el uso de pesticidas químicos, no se
descarta, a priori, la utilización de ningún tipo de agente de control (Coombs y Hall, 1998).
Metodológicamente, el control integrado de plagas puede describirse como un "proceso de toma de
decisiones" es el que, sobre la base de toda la información relevante disponible, hay que decidir qué medidas
tomar y en qué momento aplicarlas, para que el control de la plaga resulte, además de eficaz, lo más rentable
posible desde el punto de vista económico y lo menos agresivo que sea posible desde el punto de vista ambiental
(Bechinski, Mahler y Homan, 2002).
Prever en qué momento una plaga puede producir daños significativos en un cultivo es fundamental para
poder tomar una decisión con respecto a la necesidad de aplicar pesticidas (Buhler, 1996). Sin un sistema de
predicción del riesgo de daños, las opciones a las que se enfrenta un agricultor son, o bien no aplicar pesticidas, o
bien aplicarlos de forma sistemática siguiendo un criterio preventivo. La primera opción puede implicar una
pérdida de la producción si se produce una plaga. La segunda opción implica costes económicos y ambientales
que resultarán innecesarios si la plaga no se produce. Por tanto, para la utilización racional de los pesticidas se
necesita disponer de criterios que permitan determinar la necesidad de su aplicación. En la actualidad, los
programas de control integrado de numerosas especies de artrópodos y de hongos causantes de plagas en una
gran variedad de cultivos, se basan en la utilización de sistemas de predicción de riesgos (Dent, 1991; Frahm,
Johnen y Volk, 1996).
En el caso del control de plagas de gasterópodos terrestres, en los últimos años se han realizado avances,
como la comercialización del agente de control biológico P. hermaphrodita, orientados a conseguir una reducción
del uso de los molusquicidas químicos, pero el punto débil del control de plagas de caracoles y babosas continúa
siendo la falta de criterios en los que basar la decisión de aplicar los molusquicidas (Hommay, 2002; Port y Ester,
2002). Ante esta situación, los agricultores optan, por lo general, por la aplicación sistemática de molusquicidas
en sus cultivos (Bohan et al., 1997; Speiser y Kistler, 2002). Desde el punto de vista del agricultor, dicha opción se
justifica porque las babosas pueden atacar y dañar gravemente a los cultivos en cualquier época del año (Port y
Port, 1986), porque muchos cultivos, en especial en la horticultura, son extremadamente sensibles al ataque de
29

las babosas desde el momento de la plantación hasta el de la cosecha, y por el bajo coste económico de los
molusquicidas químicos convencionales (Port y Ester, 2002).
Importancia: gracias al modelo predictivo de actividad y al conocimiento del ciclo biológico de los
caracoles y babosas sabremos cuándo y cómo hay que aplicar los molusquicidas, consiguiendo con ello una gran
eficacia, saliendo beneficiados agricultores, consumidores y medio ambiente.
Progress and results assessment. Por medio de los informes semestrales y anuales, y por medio de los
controles personales que periódicamente el coordinador realiza a cada uno de los partners en el país
correspondiente.
WORK AND METHODOLOGY DESCRIPTION
It will follow the same methodology as outlined in Work Package 4, Task 4.2 for each of the tested molluscicides,
and there will be a control plot. The tested products are: non residual agrochemicals with ovicidal action and
commercial molluscicides (Methaldehude, Carabamatos, Ferramol…)
Plant’s crops damage evaluation
See Work Package 4, Task 4.2.
30

Work Package 8
Field experiments organics in key horticultural crops to evaluate the efficacy of organic Molluscicides-ovicides
and the use of plant-traps as land snails method to control pests.
PARTICIPANTS
Partners 1 (……. Man-month)
Partners 2 (……. Man-month)
Partners 3 (……. Man-month)
Partners 4 (……. Man-month)
Partners 5 (……. Man-month)
Partners 6 (……. Man-month)
Partners 7 (……. Man-month)
Partners 8 (……. Man-month)
Partners 9 (……. Man-month)
OBJETIVES
Field experiments in organic key horticultural crops to evaluate the efficacy of selected compounds with ovicidal
activity against land snail and egg-lays.
WORK AND METHODOLOGY DESCRIPTION
See Work Package 7, only change the kind of compound tested and the crop type.
31

Work Package 9
PARTICIPANTS: only Americans partners
Partners 1 (……. Man-month)
Partners 2 (……. Man-month)
Partners 3 (……. Man-month)
Partners 4 (……. Man-month)
Partners 5 (……. Man-month)
Partners 6 (……. Man-month)
Partners 7 (……. Man-month)
Partners 8 (……. Man-month)
Partners 9 (……. Man-month)
OBJETIVES
To identify improved strains of Phasmarhabditis nematodes which are more effective biocontrol agents of
larger land snail species in Hispano-America.
BACKGROUND
The use of the rhabtitid nematode Phasmarhabditis hermaphrodita (Schneider) as a biological control
agent for land snails has been proposed (Wilson, Glen & George, 1993) and a commercial product based on P.
hermaphrodita (Nemaslug , MicroBio Ltd, UK) was launched for sale in the UK in spring 1994, and later in other
European countries. Phasmarhabditis hermaphrodita has been tested successfully for biocontrol of land snails in a
number of field trials, including a range of arable and horticultural crops (Wilson et al., 1994a, 1996; Wilson, Glen,
Wiltshire & George, 1994b; Wilson, Glen, George & Hughes 1995a; Wilson, Hughes & Glen, 1995b; Ester &
Geelen, 1996; Glen et al., 1996; Iglesias, Castillejo & Castro, 2001). In some experiments, however, nematode
application did not reduce land snail damage (Wilson et al., 1995a, 1996; Speiser & Andermatt, 1996).
The strain of the nematode currently used as a biocontrol agent is well adapted to the relatively low
temperatures at which land snails are troublesome as pests in north west Europe (Glen et al.,1994a).Thus, this
strain is likely to be suitable for biocontrol of land snails in vegetable and fruit crops in northern Europe. However,
this strain may not be suitable for use in warmer regions of Latino America because it is unable to survive for
more than a few hours at temperatures greater than 25"C (Wilson et al., 1993c). It is likely that strains better
adapted to warmer regions of Latin America can be found, because P. hermaphrodita and two closely related
species from the same genus have been recorded from southern Europe (Morand, 1988).
No obstante, el elevado coste económico que suponen en la actualidad los tratamientos de control de
plagas con nematodos hace que su uso esté todavía muy restringido a cultivos de elevado valor como las plantas
ornamentales y algunas hortalizas (Grunder, 2000).
Importancia: El disponer de un nematodo zooparásito para controlar las plagas de caracoles y babosas y
que se efectivo en cultivos tropicales será muy beneficioso para la agricultura biológica.
Assessment of progress and results. Por medio de los informes semestrales y anuales, y por medio de los
controles personales que periódicamente el coordinador realiza a cada uno de los partner en el país
correspondiente.
WORK AND METHODOLOGY DESCRIPTION
STANDARD OPERATING PROCEDURE FOR PROJECT PARTNERS ISOLATING POSSIBLE NEMATODE LAND SNAILS
PARASITES
1. All land snail and soil samples collected for the identifying purpose of possible new nematode parasites will be
labeled with the following essential information, which it will be provided for every new strain of nematodes
found.
* Nearest place name, nearest main town, and Country
* Latitude and longitude
* Land snail species (if sample is from a land snail)
32

* Number of each species (if sample is from more than one land snail)
* Soil type
* Crop or habitat
* Recollection Date
* Recollection method
* Recent and current weather
* Number of subcultures (if any) since nematodes were isolated from infective land snails
2. At all times, land snails suspected of being parasited with nematodes (swollen mantle symptom) should be kept
in individual Petri dishes lined with moist filter paper at below 20 ºC (recommended 15 ºC). Land snails should be
kept like this until nematodes have reproduced within/on the cadaver.
3. Once nematodes are seen crawling on the land snail cadaver (this is usually about 7 days after infection; at this
point adult and some juvenile nematode stages should be present), the land snail re++++++++mains should be
transferred onto either an extraction tray or a modified White trap and kept at the same temperature as above
for several days.
Extraction tray method (also known as modified Baermann tray or Whitehead tray) (in EU first 6-month report,
previously referred to as double tray/milk filter technique):
This method is frequently used for extracting plant-parasitic nematodes from root material Whitehead and
Hemming, 1965; Southey, 1986). It consists of a coarse nylon sieve (1 mm gauze) which has supports fixed to the
bottom lifting it about 1 cm up. It is lined with a single layer of tissue paper or milk filter to prevent waste material
getting through. The sieve is then placed in a tray with water just reaching the bottom of the sieve. Infected land
snails can be placed on the sieve. Nematode stages will migrate out of the land snail cadaver into the Water, with
the filter preventing unwanted land snail remains contaminating the water. Trays should be left for several days
allowing all nematodes to emerge from the land snail remains. Nematodes collected in the water can be
harvested every other day or so, with the tray being refilled with fresh water.
White trap: The modified White trap it is a standard way of extracting and collecting entomopathogenic
nematodes from infected insect larvae (White, 1927; Woodring and Kaya, 1988). It is probably also a better
method of collecting the infective juveniles of land snail-parasitic nematodes than using extraction trays. It
consists of a small container (9 cm diameter x 5 cm deep) in which an inverted 5 cm diameter, 2 cm deep Petri
dish bottom is placed. A shallow layer of water (1 cm deep) is added to the container. A filter paper is then draped
on the Petri dish platform so that it comes into contact with the water. Infected land snails are then placed onto
the moist filter paper on the platform. Once the nematode has completed its life cycle, infective juveniles will
start to emerge from the land snail remains and migrate across the moist filter paper into the water after about
14 days (or longer depending on nematode species, level of infection and temperature). Nematodes can be
harvested every other day or so, until nematodes are no longer found in the water. The container should be
refilled with fresh water after every harvest.
Ideally, only one infected land snail should be placed in each extraction tray or White trap. If this is impossible
(e.g. because large numbers of infected land snails have been collected at the same time), then each tray should
contain land snails of one species, collected from the same site at the same time.
4. Infective juveniles collected should be cleaned before storage. The nematode suspension is poured in a small
(100 ml) beaker and left undisturbed for about 30 min, allowing nematodes to settle onto the bottom. The
supernatant is then carefully decanted, leaving behind a concentrated nematode suspension. The beaker should
then be refilled with fresh well-oxygenated tap water (or preferably sterile distilled water) and the processes are
repeated 3 to 4 times or until the suspension appears clear. Nematodes can then be stored in shallow containers
with breathing holes, the suspension being only approximately 0.5 cm in depth. Storing nematodes in a large
surface-to-volume ratio ensures sufficient oxygen availability. The suspension should ideally have a concentration
of no more than approximately 5000/ml and be stored at 5 ºC. Nematode viability should be checked regularly
(after room temperature acclimatization). Nematodes should be hatched again after several months or when a
large proportion of nematodes start to die.
33

5. A live sample of each isolated nematode will be sent by Courier, to Partner 1 for specific identification.
Nematodes will be sent as infective juveniles in tap water in a part-filled container, with a large ratio of air to
water (20 volumes of air to 1 volume of water) for culturing and confirmation of species identity.
34

CALL: FP7-KBBE-2010-4
Proposal full title: Novel strategies for integrating land snail pests control of agricultural
crops in Europe, with projection to Latin-America
Proposal acronym:
Short name?:
Type of funding scheme:
Work programme topics
addressed:
LAND SNAIL PEST CONTROL, LAND SNAIL CONTROL, CONTROLLING LAND
SNAILS
Land snail’s life cycle as pest control core
Collaborative Project. CALL: FP7-KBBE-2010-4
Food, Agriculture and Fisheries, and Biotechnology
Area: 2.1.2
Topic number: KBBE.2010.12-05
Name of the coordinating person: Dr José Castillejo Murillo
Departamento de Zoología y Antropología Física. Facultad de Biología.
Universidad de Santiago de Compostela. E-15782 Santiago de Compostela.
La Coruña. Galicia. España.
List of participants
Mobile Phone: + 34 654 969 784
Tel: + 34 981563100
Fax: + 34 981 596904
E-mail: jose.castillejo@usc.es
Participant number Participant organitation name Country
1 (Coordinator) Universidad de Santiago de Compostela Spain
2
3
4
5
6
7
8
9
10
11
12
13
14
15

Gantt Chart showing the timing of the different WPs and their components.
Work
Packge
Nº
WP.1
WP.2.
WP.3.
WP.4.
WP.5.
Task
Nº
Task 4.1.
Task 4.2.
Task 4.3.
Task 4.4.
Task 5.1.
Task 5.2.
Task 5.3.
Work Packages Title
Land Snail Biological Cycle. The size, structure and
dynamics of their
populations
Participant No. Participant
organism name
All partners
Land Snails diet composition. Trap plants All partners
Statistical Model to predict land snails activity All partners
Bio pesticides, Bio Molluscicides, Bio Ovicides
Plants Extracts. Laboratory test on paper filter
Standard soil
All partners
Mini plots tests on horticultural soil All partners
Collateral effects on soil fauna All partners
Chemicals analysis to search the active principle of
plant extracts with mulliscicide and ovicide activity
Laboratory test to find non residual agrochemicals
with ovicidal activity
Field experiments on conventionally horticultural
crops to evaluate the efficacy of selected
agrochemical as
molluscicide-ovicide
Field trials to evaluate the collateral effects on soil
Fauna and border effect on wild land snails of
ovicide agrochemicals.
Sygenta
Company and
USC (Spain)
All partners
All partners
All partners
First Year Second Year Third Year
1 2 3 4 5 6 7 8 9 1
0
1 2 3 4 5 6 7 8 9 1
0
1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
0
1 2 3 4 5 6 7 8 9 1
0
1 2 3 4 5 6 7 8 9 1
0
1 2 3 4 5 6 7 8 9 1
0
1 2 3 4 5 6 7 8 9 1
0
1 2 3 4 5 6 7 8 9 1
0
1
1
1
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1
2

WP.6.
WP. 7
WP. 8
Task 6.1.
Task 6.2.
Task 6.3.
Task 6.4
Manure Laboratory test on land snail eggs for
accurate
Ovicidal concentration
Field experiments to evaluate the efficacy of cow and
pig manure as slug eggs control for key organic
horticultural crops
Field analysis to investigate the collateral effect
on soil fauna and border effect on wild land
snails of cow and pig manure
Field experiments to use tramp-plants as deterrent
method to protect organic horticultural crops alone
and in combination of cow and pig manure
Field experiments in conventionally crops to evaluate
the efficacy of ovicide agrochemicals alone and in
combination with other commercial molluscicides
Field experiments in organics horticultural crops to
evaluate the efficacy of organic molluscicidesovicides
and the use of plant-traps
WP. 9 To identify improved strains of Phasmarhabditis
nematodes which are more effective biocontrol
agents of larger slug species in Hispano-America.
All partners
All partners
All partners
All partners
All partners
All partners
Only Latino
America
Parteners
1 2 3 4 5 6 7 8 9 1
0
1 2 3 4 5 6 7 8 9 1
0
1 2 3 4 5 6 7 8 9 1
0
1 2 3 4 5 6 7 8 9 1
0
1 2 3 4 5 6 7 8 9 1
0
1 2 3 4 5 6 7 8 9 1
0
1 2 3 4 5 6 7 8 9 1
0
1 2 3 4 5 6 7 8 9 1
0
1 2 3 4 5 6 7 8 9 1
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1
1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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1 2 3 4 5 6 7 8 9 1
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2
1
2
1
2

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
Packge
Nº
WP.1
WP.2.
WP.3.
WP.4.
WP.5.
Task
Nº
Task 4.1.
Task 4.2.
Task 4.3.
Task 4.4.
Task 5.1.
Task 5.2.
Task 5.3.
Work Packages Title Hours per day
Land Snail Biological Cycle.
the size, structure and dynamics of their
populations
Land Snails diet composition. Trap plants
Statistical Model to predict land snails activity
Bio pesticides, Bio Molluscicides, Bio Ovicides
Plants Extracts. Laboratory test on paper filter
Standard soil
Mini plots tests on horticultural soil
Collateral effects on soil fauna
Number of
days per
month
Number of
month
Number of
person
participating
Total Hours per
WP or Task
8 5 24 2 1920 14.2
12 5 24 2 2880 21.3
12 5 24 2 2880 21.3
4 10 30 2 2400 17.7
2 10 24 2 960 7.1
2 10 18 2 720 5.3
Chemicals analysis to search the active principle of
plant extracts with mulliscicide and ovicide activity 2 10 18 2 720 5.3
Laboratory test to find non residual agrochemicals
with ovicidal activity 4 10 25 2 1920 14.2
Field experiments on conventionally horticultural crops to
evaluate the efficacy of selected agrochemical as
molluscicide-ovicide
4 10 18 2 1440 10.6
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
Person-month
Hours/135
2.6

WP.6.
WP. 7
WP. 8
WP. 9
Task 6.1.
Task 6.2.
Task 6.3.
Task 6.4
Manure Laboratory test on land snail eggs for accurate
Ovicidal concentration
Field experiments to evaluate the efficacy of cow and
pig manure as slug eggs control for key organic
horticultural crops
Field analysis to investigate the collateral effect
on soil fauna and border effect on wild land
snails of cow and pig manure
Field experiments to use tramp-plants as deterrent
method to protect organic horticultural crops alone
and in combination of cow and pig manure
Field experiments in conventionally crops to evaluate
the efficacy of ovicide agrochemicals alone and in
combination with other commercial molluscicides
Field experiments in organics horticultural crops to
evaluate the efficacy of organic molluscicidesovicides
and the use of plant-traps
To identify improved strains of Phasmarhabditis
nematodes which are more effective biocontrol
agents of larger slug species in Hispano-America.
2 10 18 2 720 5.3
2 10 24 2 960 7.1
1 10 18 2 360 2.6
1 5 24 2 240 1,7
4
10 24 2 1920 14.2
4 10 24 2 1920 14.2
1 10 36 2 720 5.3
TOTAL person-months 209.7

Deliverables
1. Contract deliverables
At the end of each year a progress report will be produced by each of the individual participants in which research
methods and results achieved will be included and related to the project milestones. After 18 months (half the duration
of the project) a project mid-term review report will also be produced following a mid-term review meeting. This midterm
report will include input from all partners and will summarise overall progress and needs for the remainder of the
project duration. After three years (end of project) a final report will be produced.
2. Technical deliverables
(a) The introduction into vegetable and fruit crops of a novel strategy for land snails control, combined where
appropriate with novel low-toxicity agrochemicals with bio-ovividal activity, as a replacement for current chemical
which give inadequate control, are hazardous to pets, wildlife and beneficial invertebrates and cause concerns because
of residues in food crops.
(b) Novel methods of bio-molluscicides and bio-ovicides control of slugs in vegetable crops.
(c) Effective integrated packages of crop management, biopestices methods and, where appropriate, low-chemical
methods of protecting vegetable crops from slug damage.
Each year, technologies developed in this project will be transferred via extension services and consultants, to
vegetable growers. Further development of results will be undertaken by Partner 2 (a SME), so as to introduce the novel
bio-mollusicicide agent into the vegetable growing sector of the European horticultural industry. For plan strects, official
registration of the product will be required and appropriate industrial partners will be involved to do this. At the end of
the project results will be further developed to provide practical advice for vegetable growers.
Table 1.3 b: Deliverables List
Del.nº Deliverable name WP nº Nature
1.1
1.2
1.3
2.1
2.2
3.1
3.2
Conocer el ciclo biológico de las especies plaga en
función de la fenología del cultivo y su situación
geográfica con el objeto de introducir medidas
preventivas
Conocer por medio del estudio del tamaño y estructura
de la población la magnitud de los daños que las
babosas y caracoles causan en los cultivos
Saber por medio de la dinámica de las poblaciones de
caracoles y babosas en que fases de su desarrollo son
más dañinos para los cultivos
Conocer las plantas más apetecidas por los caracoles y
babosas por medio del estudio de sus contenidos
estomacales.
Obtener la base científica para desarrollar la estrategia
del uso de plantas-trampas para proteger los cultivos
Conocer los periodos de actividad en cada una de las
zonas de estudio en función de las condiciones medio
ambientales y la fenológicas
Desarrollar un Modelo Estadístico que nos prediga los
periodos de actividad de las especies de gasterópodos
terrestres plaga
Dissemi
nation
level
WP. 1 R PU
WP.1 R PU
WP.1 R PU
WP.2 R PU
Deliverable
date.
Months
Partial: 12
Final: 24
Partial: 12
Final: 24
Partial: 12
Final: 24
Partial: 12
Final: 24
WP.2 R PU Final: 24
WP.3 R PU
WP.3 R PU
Partial: 12
Final: 24
Partial: 24
Final: 36

3.3
3.4
4.1
4.2
4.3
5.1
5.2
5.3
6.1
6.2
6.3
Tener información de cuál es el momento más
adecuado para aplicar un tipo concreto de molusquicida
o emplear una estrategia de control idónea
Proporcionarle al agricultor información para que
desarrolle una estrategia preventiva con la que se
adelante al ataque de la plaga, use menos
molusquicidas y consiga una mayor eficacia
Conocer plantas de las que se puedan obtener
biopesticidas con acción molusquicida y ovicida para
que en futuro reemplacen a los molusquicidas
tradicionales, o bien con ellas se puedan hacer abonos
verdes que puedan ser usados en cultivos ecológicos
como controladores de las plagas de caracoles y
babosas. (WP. 4)
Poner en mano de la industria de agroquímicos nuevas
vías de control de las plagas de caracoles y babosas por
medio de Biomolusquicidas y Bio-ovicidas obtenidos a
partir de extractos de plantas. The possibility to
introduce into key horticultural crops of a novel plant
extract as biocontrol agent for land snails control
Saber que efectos colaterales puede tener el uso de
biomolusquicidas y bio-ovicidas sobre la fauna edáfica
y de zonas colindantes (WP.4)
Development of novel techniques for low-toxicity
chemical control of land snail eggs - the stage in the
pest life cycle against which no current controls are
available
Proporcionarle al agricultor información de cómo usar
los agroquímicos que habitualmente emplea en los
cultivos tradicionales le puedan servir para controlar las
plagas de los gasterópodos terrestres
Proporcionarle información al agricultor de los efectos
colaterales que sobre la fauna pueden tener los
molusquicidas-ovicidas que se proponen usar en
cultivos tradicionales
Proporcionarle al agricultor ecológico información de
cómo puede usar los abonos naturales para controlar
las plagas de gasterópodos terrestres
Indicar al agricultor ecológico que tipos de plantas
puede usar para disuadir a los gasterópodos terrestres
de que no ataquen a los cultivos
Saber que efectos colaterales puede tener el uso de
abonos orgánicos a concentración ovicida sobre la
fauna edáfica y de zonas colindantes
WP.3 R PU
WP.3 R PU
WP.4
WP.4
R PU
R PU
R PU
WP.5 R PU
WP.5 R PU
WP.5 R PU
WP.6 R PU
WP.6 R PU
WP.6 R PU
Partial: 12
Partial: 24
Final: 36
Partial: 24
Final: 36
Partial: 24
Final: 36
Partial: 24
Final: 36
Partial: 24
Final: 36
Partial: 24
Final: 36
Partial: 24
Final: 36
Partial: 24
Final: 36
Partial: 24
Final: 36
Partial: 24
Final: 36
Partial: 24
Final: 36
7.1 Proporcionarle al agricultor convencional información WP.7 R PU Partial: 24

8.1
9.1
9.2
5&6.1
5&6.2
5&6.1
4&5&6.
1
6&9.1
7&8.1
All.1
sobre la forma de usar los distintos molusquicidas solos
o en combinación, y sobre su eficacia de estos
dependiendo del tipo de cultivo, del tipo suelo y de las
especie de gasterópodo terrestre que son plaga en una
zona concreta
Proporcionarle al agricultor ecológico información
sobre la forma de usar los distintos biomolusquicidas
solos o en combinación, y sobre su eficacia de estos
dependiendo del tipo de cultivo, del tipo suelo y de las
especie de gasterópodo terrestre que son plaga en una
zona concreta
Conocer si existen posibles nematodos zooparásitos
que se puedan ser usados en el control biológico de las
plagas de caracoles y basas en cultivos agrícolas
The possibility to introduct into key horticultural crops
of a novel nematode biocontrol agent for land snails
Greater uptake of environmentally favourable
integrated crop production systems, as a result of
reduced risk of land snail damage
Improved food safety, resulting from reduced levels of
molluscicide residues in harvested produce
Effective integrated packages of control measures for
protecting conventionally grown horticultural crops
from land snails damage
Improved environmental safety, resulting from reduced
reliance on chemical molluscicides
Effective integrated packages of control measures for
protecting organically grown horticultural crops from
land snail damage
Technology transfer through meetings of growers,
practical demonstrations, extension services, trade
shows and articles in the horticultural press
Brochure with two parts, or separate brochures
describing integrated packages of control measures for
conventional and organic horticultural producers and
Web links for specific advices.
WP.8 R PU
WP.9 R PU
Final: 36
Partial: 24
Final: 36
Partial: 24
Final: 36
WP.9 R PU Final: 36
WP.5&6 R PU Final: 36
WP.5&6 R PU Final: 36
WP.5&6 R PU Final: 36
WP.
4&5&6
WP.
6&9
(WP.
7&8
R PU Final: 36
R PU Final: 36
R PU Final: 36
(WP. all) R PU Final: 36

Milestones
The project milestones for the nine individual work tasks are shown in Table 1.3 c
Table 1.3 c: Annual milestones for the 9 Work Packages proposed.
Milestone
number
Milestone name Work
Package(s)
Dinámica, estructura y tamaño de la población of land
snail pest. First year on field sampling
Complete field study of land snails populations to
establish its dynamic, structure and size.
Establish best methods to find out the land snails fife
cycle.
involved
Espected
Data
Means of
verification
WP. 1 12 Field survey
complete and
report
WP.1 24 Field survey
complete and
report
WP. 1 12 Field survey
complete and
Wp.1 24
report
Field survey
Life cycle, complete field observation and sampling
complete and
report
WP. 2 12 Field survey
Establish best methods to find out the land snails diet.
complete and
report
Complete field study of land snails diet. WP.2 24 Field survey
complete and
report
First draft of the Statistical Model to predict the land WP. 3 12 Complete
snails activity
laboratory
processing and
report
Second draft of the Statistical Model to predict the land WP. 3 24 Complete
snails activity
laboratory
processing and
report
Definitive Statistical Model to predict the land snails
activity in every zone studied
WP. 3 36 Report
Select potential plants for extracting compound as bio- WP.4 12 Laboratory survey
molluscicide
complete and
report
Select better compound for direct contact on filter WP.4 12 Laboratory survey
paper. Results of laboratory tests
complete and
report
Select better compound for mini plots testing and better WP.4 24 Field survey
methods of application.
complete and
report
Field test of best compounds for determinate the effect WP.4 24 Field survey
on non-target soil animals
complete and
report
Select better agrochemical based on laboratory tests WP. 5 12 Laboratory survey
complete and
report
Complete small-scale field trials on agrochemicals WP. 5 24 Field survey
complete and
report
Select cow and pig manure concentration based on WP. 6 12 Laboratory survey
laboratory tests
complete and

eport
Complete small-scale field trials on manure WP. 6 24 Field survey
complete and
Parcial estudio de la actividad de los gasterópodos
terrestres plaga. Refine use of best methods.
report
WP. 3 12 Field survey
complete and
report
Complete second year of activity WP. 3 24 Field survey
complete and
report
Complete third year of activity, definitive model WP. 4 36 Field survey
complete and
Evaluar la eficacia de los ovicidas en conventionally crops
Complete small-scale field trials
Evaluar la eficacia de los purines como ovicidas en
organic crops. Complete small-scale field trials
Conocer los efectos colaterales de los molusquicidas
ovicidas, partial trials.
Partial and Complete field test on side effects.
Investigate effects on no-target soil fauna.
Comprobar la capacidad de las plantas trampa para
proteger los cultivos
Comparar la eficacia en conventionally crops de los
molusquicidas ovicidas frente a los molusquicidas
tradionales. Determine best way to combine with others
molluscicides.
Comparar la eficacia de los ovicidas orgánicos frente a
controles biológicos. Determine best way to combine
with others molluscicides.
Búsqueda de un nematodo Phasmarhadities autóctono
para usarlo en el control biológico. Select the best
methods based on laboratory tests.
Devise integrated packages for further development on
conventional crops
Devise integrated packages for further development on
organic crops
report
WP. 5 24 Field survey
complete and
report
WP. 6 24 Field survey
complete and
report
WP. 5 &6 24 Field survey
complete and
report
WP. 5&6 24 and 36 Field survey
complete and
report
WP. 7 24 Field survey
complete and
report
WP. 7 24 Field survey
complete and
report
WP. 8 24 Field survey
complete and
WP. 9 12, 24 and
36
WP. 1,2,3,
4,5,6,7 y 8
WP. 1,2,3,
4,5,6,7 y 8
report
36 Report
Field survey
complete and
report
36 Report

RELATIONSHIP TASKS and ParticipantS
WP 9
NEMATODO ZOOPARASITIC
Phasmarhadities
Participant: Latino America
WP 1
LAND SNAIL’S BIOLOGICAL
CYCLE
Participant 1, 2 ,3, 4, 5, 6, 7,
8
WP 7 & 8
FIED EXPERIMENTS
CONVENTIONALLY,
ORGANICS CROPS
Participant 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
WP 6
PIG AND COW MANURE
AS BIO OVICIDES 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 5
BIO OVICIDAS
AGROCHEMICALS TESTS
WP 4
BIOMOLLUSCICIDES TESTS
COLLATERAL EFFECTS
Participant 1, 2 ,3, 4, 5, 6, 7,
8
Participant 1, 2 ,3, 4, 5, 6, 7.
8

RELATIONSHIP TASKS and Participants
WP 9
NEMATODO ZOOPARASITIC
Phasmarhadities
Participant: Latino America
WP 1
LAND SNAIL’S BIOLOGICAL
CYCLE
Participant 1, 2 ,3, 4, 5, 6, 7,
8
WP 7 & 8
FIED EXPERIMENTS
CONVENTIONALLY,
ORGANICS CROPS
Participant 4, 5, 6, 7, 8
WP 2
DIET COMPOSITION
TRAP PLANTS
Participant 1, 2 ,3, 4, 5, 6, 7,
8
intercommunication
WP 6
PIG AND COW MANURE
AS BIO OVICIDES 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 5
BIO OVICIDAS
AGROCHEMICALS TESTS
WP 4
BIOMOLLUSCICIDES TESTS
COLLATERAL EFFECTS
Participant 1, 2 ,3, 4, 5, 6, 7,
8
Participant 1, 2 ,3, 4, 5, 6, 7.
8

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

Table 2 Schedule of meeting to be held and reports to be produced
MEETING Due Date
Initial planning meeting, Santiago de Compostela, Spain
December 2010
(at the beginning of Project)
Progress review & planning meeting, Norwich
December 2011
Progress review & planning meeting, Lithuania
Final review meeting, Argentina, Ireland
MEANS OF COORDINATOR VERIFICATION
First travel to verification, Europe/Hispano America
Second travel to verification, Europe/Hispano America
Third travel to verification, Europe/Hispano America
SIX MONTH PROGRESS REPORT
First Six-month Progress Report
Second Six-month Progress Report
Third Six-month Progress Report
ANNUAL PROGRESS REPORT
First Annual Progress Report
Second Annual Progress Report
Third Annual Progress Report
December 2012
December 2013
May 2011
October 2011
June 2012
June 2011
June 2012
June 2013
December 2012
December 2013
December 2014
FINAL REPORT March 2014
Brochure(s) and Web page for extension services, consultants and/or growers March 2014

SUMMARY OF STAFF EFFORT
Participant
no./short name
No. 1 USC-ES
No. 2
No. 3
No. 4
No. 5
No. 6
No. 7
No. 8
No. 9
No. 10
No. 11
No. 12
WP 1 WP 2 WP 3 WP 4 WP 5 WP 6 WP 7 WP 8 WP 9
Total person
month

Project: Partner 1 – Universidad de Santiago de Compostela, Spain
Table – Estimated Breakdown of the Total Allowable Cost
ALLOWABLE COST AMOUNT IN EUROS
LABOUR 237600 €
TRAVEL, SUSBSISTENCES AND MEETINGS 118200 €
DURABLE EQUIPMENT 116000 €
CONSUMABLES 40000 €
EXTERNAL ASISTANCE
COMPUTING & OTHER COST 40000 €
OVERHEADS 20% 110360 €
TOTAL Requested from EU
Rounded to
a) A detailed list of personnel to the execution of the research (man/month)
Postdoctoral Research Scientist (Group 1) ………………. 36 man/month
Graduate Research Scientist (Group 2) ……………………. 36 man/month
Official Salary Scales (See attached documents)
Group 1, Postdoctoral Research Scientist: 144000 Euros/36 months before tax
108000 Euros/36 months after tax
Group 2, Graduate Research Scientist: 93600 Euros/year before tax
72000 Euros/year after tax
b) Idem for “external assistance”
SHARED COST PROJECT
662160 €
663000 €
144000 €
93600 €
TOTAL 237600 €
TOTAL

c) A list of meeting for planning and travel and subsistence
COST Euros
3 Meetings for planning & coordination 36000 €
Land snails sampling in Spain (see below for sampling) 37800 €
3 Hispano America/Europa extra trip to verification and adviser 44400 €
Details of Meeting for planning & coordination
(3 staff x 7 days x 3 meetings). Argentina, Bolivia y Costa Rica
Precio billete avión ida/vuelta: 3000 Euros/persona x 3 personas x 3 viajes= 27000 €
Alojamiento hotel: 100 Euros/noche persona x 5 noches x 3 personas x 3 viajes = 4500 €
Manutención: 70 Euros/day persona x 7 días x 3 personas x 3 viajes = 4500 €
TOTAL: 36000 Euros
TOTAL 118200 €
Details of land snail sampling in Spain
Sampling for land snail density and activity in crops, monthly: 2011, 2012 y 2013.
12 annual trips, three zones visited from 5 characteristic crops in each zone. Each trip will involve three
people for a total 3 days per trip.
Year 2011: 12 trips x 3 people x 3 days x (60 € hotel + 50 € subsistence) = 11800 €
Year 2012: 12600 €
Year 2013: 13400 €
TOTAL: 37800 €
Extra Hispano American trip for verification and adviser
3 extra trips to Hispano America/Europe to check in situ project’ development and correct possible
mistakes.
Trip for 2 people and 20 days lenth.
Return Plane ticket: 4000 €/trip/person = 24000 €/3 return ticket/2 people
Hotel: 20 nights x 3 trips x 2 people x 100 € Room = 12000 €
Subsistence: 20 days x 3 trips x 2 people x 70 € subsistence = 8400 €
TOTAL: 44400 €
d) A list of durable equipment
2 cámaras FITOCLIMA D1200PLH con control total del medio interior: temperatura,
humedad, iluminación, aireación.
Modelo ARALAB D1200PLH y precio: 27000 €/unit
Invertido de bajos aumentos mod. Ix51. Optica de campo claro. Salida triocular lateral
para foto/tv. Con sistema de fotografía y video recorder
Inverted Stereo zoom microscope, with viseLed White LED illumination system Digital
Camera
54000 €
30000 €
10000 €
Digital System to Store activity dates, images and habitat variables 9000 €
5 Digitals AXIS IP cameras to control activity of field land snails with remote control and
Wifi connection
13000 €
TOTAL 116000 €

e) Consumables. Details of others cost
Computing 4000 €
Field sampling facilities 6000 €
Field experiments 10000 €
Laboratory test facilities 10000 €
Laboratory test consumables 10000 €
TOTAL 40000 €

DIRECCIONES E-MAIL Proyecto de la Unión Europea 2009
EUROPA
Dr. José Castillejo Murillo
Departamento de Zoología
Facultad de Biología
Universidad de Santiago de Compostela
15782 Santiago de Compostela
La Coruña. España
E-Mail: jose.castillejo@usc.es
--------------------------
Dr. David M Glen
Styloma Research & Consulting
Phoebe, The Lippiatt, Cheddar, Somerset, BS27 3QP, UK
Tel: +44 (0)1934 743277
Mobile: +44 (0)7815 624971
David Glen davidmglen@btopenworld.com
--------------------------
Dr. André Chabert
ACTA. 4 place Gensoul
69287 Lyon Cedex 12
France
E-mail: andre.chabert@acta.asso.fr
--------------------------
Dr Albert Ester
PAV. Edelhertweg
P.O. Box 430
8200 AK Lelystad. The Netherlands
E-mail: Albert Ester albert.ester@wur.nl
--------------------------
Dr. A.S.H. Breure.
National Museum of Natural History
P.O. Box 9517. Leiden. Holanda ( The Netherlands)
Phone: +31 71 56 78 552 Room Number D 03.47
E-mail: breure@naturalis.nnm.nl
--------------------------
Dr. BERNHARD HAUSDORF
Zoologisches Institut und Zoologisches Museum der Universität Hamburg,
Martin-Luther-King-Platz 3, D-20146 Hamburg, Germany
E-mail: hausdorf@zoologie.uni-hamburg.de
--------------------------
David Glen davidmglen@btopenworld.com
Albert Ester albert.ester@wur.nl
Brigitta Grimm brig@nhm.ac.uk
Gordon Port gordon.port@ncl.ac.uk
Bill Bailey bbailey@man.ac.uk
Bernhard Speiser Bernhard.speiser@fibl.ch
Gerard Hommay hommay@colmar.inra.fr
Andre Chabert andre.chabert@acta.asso.fr
-------------------------------------
Syngenta Crop Protection
Syngenta Crop Protection UK Ltd.
CPC4 Capital Park. Fulbourn
Cambridge CB21 5XE. UK
E-mail: customer.services@syngenta.com

AMERICA
CHILE
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
E-mail: sletelier@mnhn.cl
---------------------------------
Dr. Carlos Fernández
Director Regional
INIA La Platina
Santa Rosa, 11610 – La Pintana
Santiago
Chile
E-mail: cfernandezb@inia.cl
--------------------------------------------
Dra. Marta Isabel Abalos Romero
Directora Ejecutiva INFOR
Dirección: Santiago, Chile
Correo-e: mabalos@infor.cl
------------------------------------------
Dr. Jaime del Canto
Subdirector Regional de Administración y Finanzas
INIA La Platina
Santa Rosa, 11610 – La Pintana
Santiago. Chile
E-mail: jdelcanto@inia.cl
------------------------------------------------
Dr. Leopoldo Sánchez Grunert
Director Nacional INIA
Dirección: Santiago, Chile
Correo-e: lsanchez@inia.cl
----------------------------------------------------
Dr. Marcos Gerding
Instituto de Investigaciones Agropecuarias (INIA)
Centro Regional de Investigación Quilamapu
Centro Tecnológico de Control Biológico
Chillán, Chile.
Tel: +56 - 42 - 209705
Fax +56 - 42 - 209720
E-mail: mgerding@inia.cl
-------------------------------------------
Centro Tecnológico de Control Biológico
Avda. Vicente Méndez 515.
Chillán – Chile.
Tel. (56- 42)- 209 700
Fax. (56 – 42) – 209 720
E-mail: controlbiologicochile@inia.cl
----------------------------------------------

CHILE
Facultad de Ciencias Biologicas
Pontificia Universidad Católica
Avenida Bernardo O'Higgins 340 ó Portugal 49
Santiago, Chile Casilla 114-D
Telefóno: (56 2) 354 2673
Fax: (56 2) 354 2369
E-mail: decanato@bio.puc.cl
Universidad de Concepción Chile
Facultad de Ciencias Biológicas, Universidad de Concepción, Chile
Casilla 160 C Fono: 204508,
Mail: csbiolog@udec.cl

ARGENTINA
Vernavá, María Natalia
Avenida Alem 706 - 4 C,
(8000) Bahía Blanca. Argentina
E-mail: nvernava@uns.edu.ar
E-mail: mnvernava@hotmail.com
--------------------------------------------
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
E-mail: acastillo@balcarce.inta.gov.ar

URUGUAY
Dr. Dan Piestun Malinow
Dr. Mario Garcia Petillo
Instituto Nacional Investigaciones Agropecuarias
Ingeniero Agrónomo, Dr.
Andes 1365 P.12
Teléfono:9020550 (oficina)
E-mail: dpiestun@inia.org.uy
E-mail: mgarcia@dn.inia.org.uy
E-mail: mallegri@inia.org.uy
Área Ciencias Agrarias
Facultad de Agronomía
Dirección: Av. Garzón 780 - C.P.: 12.900
Tel.: (598 2) 3597191-94 Fax: 3590436
http://www.fagro.edu.uy/
Montevideo - Uruguay
Regional Norte
Dirección: Rivera 1350
Tel.: (598 73) 34816 / 20412 / 29149 / 22154 / 26603 Fax: (073) 20412
Dirección: Uruguay 1375
Tel.: (598 73) 20108
Correo electrónico: web@unorte.edu.uy
http://www.unorte.edu.uy/
Salto - Uruguay
Estación Experimental “Dr. Mario Cassinoni” (E.E.M.A.C.) - Paysandú
Ruta 3 - km 363 - C.P.: 60.000
Tel.: (598 72) 41282 / 02250 / 02259
Telefax: (598 72) 27950
http://www.fagro.edu.uy/~eemac/
Paysandú - Uruguay
DECANATO (Universidad de la República de Uruguay)
de la Facultad de Agronomía
Ing. Agr., Agrícola-Ganadera, FA-UDELAR, 1974
M.Sc., Manejo de Suelos, Iowa State Univ., 1986
Ph.D., Manejo de Suelos, Iowa State Univ., 1991
e-mail: decanato@fagro.edu.uy
Departamento de Producción Vegetal
Encargada de Dirección: Ing. Agr. (PhD) Milka Ferrer
e-mail: mferrer@fagro.edu.uy
Dan Piestun Malinow
INIA Uruguay: dpiestun@inia.org.uy

BOLIVIA
Dra. Elva Terceros Cuellas
Directora General INIAF
La Paz, Bolivia
Correo-e: elva.terceros@iniaf.gov.bo
----------------------------------------------------
Víctor Hugo Cardoso
Secretarío Ejecutivo
PROCIANDINO
Av. Palca y Calle 54 Zona Chasquipampa -
La Paz. Bolivia
Teléfono: (591-292772145
Fax: (591-2) 2773399
E-mail: victor.cardoso@iica.int.bo
Facultad de Ciencias Agrícolas, Pecuarias, Forestales y Veterinarias
“Martín Cárdenas”
Casilla No. 4894 Teléfonos: 4333808 - 4329666
Fax: (591)(4) 4762385
Cochabamba-Bolivia
E-mail: postmaster@agr.umss.edu.bo (devuelto)
E-mail: agro@agr.umss.edu.bo (devuelto)
marioescalier@yahoo.com
cocamorante.mario@gmail.com
cocamario@hotmail.com
--------------------------------------
Instituto Boliviano de Biología de Altura (IBBA)
Calle Claudio Sanjinés s/n (Miraflores)
Telf.: (591-2) 2242064 / (591- 2) 2242059
Fax :(591-2) 2221418 Casilla postal: 641
email: Ifisibba@fdm.umsalud.edu.bo (Devuelto)
La Paz - Bolivia

PERÚ
Dr. Enrique La Hoz Brito.
Director General de Investigación.
DIA - INIAAv. La Molina (Ex Av. La Universidad) No. 1981 Lima 1
Perú
E-mail: elahoz@inia.gob.pe
elahoz@inia.gob.pe
Enrique La Hoz
"Risi Carbone, Juan Jose M."
----------------------------------------------
Dra. Rina Ramírez
Museo de Historia Natural. Universidad Nacional Mayor de San Marcos,
Lima – Perú. Apartado 14-0434, Lima14, Perú
E-mail: rina_rm@yahoo.com
--------------------------------------------------
Universidad Nacional Agraria La Molina
Facultad de Agronomía. Perú
Marlene Aguilar Hernández MARLENE AGUILAR
----------------------------------------------------------
Universidad Nacional de Trujillo-Peru
Profesor de zoología y entomología
AURELIANO RAMIREZ CRUZ (Dr .Martin Delgado)
E-mail: aure_15@hotmail.com
Nombre : Andrés V. Casas Díaz
Categoría : Profesor Principal, dedicación exclusiva
Especialidad: Olericultura
Título : Ing. Agr. Mg.Sc.
Cursos: Olericultura Especial (T), (P)
Fisiología y Manejo Postcosecha (P)
correo : cda@lamolina.edu.pe
Màs información: Programa de Hortalizas
Nombre: Julio Toledo Hevia
Categoría : Profesor Principal, dedicación exclusiva
Especialidad: Olericultura y Manejo Postcosecha
Titulo :Ing. Agr. Ph. D.
Cursos: Fisiología y Manejo Postcosecha (T)
Fisiología de Cultivos Hortícolas (T), (P)
correo : jtoledo@lamolina.edu.pe

ECUADOR
Universidad Central de Ecuador
Centro de Coordinación Académica de Biología
Relaciones internacionales: rel_int@ac.uce.edu.ec
Instituto de Investigaciones Agrícolas
Facultad de Ciencias Agrícolas ( Interesado)
Universidad Central del Ecuador
Narciza Yar (FUNCIONARIA DE LA FACULTAD DE CIENCIAS AGRÍCOLAS)
E -mail: narcizayarm@hotmail.com
Universidad Central de Ecuador
Vicerrector Académico y de Investigación
Dr. Jorge Luciano Arroba Rimassa
E-mail: jarroba@ac.uce.edu.ec
Universidad Agraria del Ecuador
Director
E-mail: elmisionero@uagraria.edu.ec
Universidad Nacional de Loja (Ecuador)
DIRECTORES DE LAS ÁREAS
Área Agropecuaria y de Recursos Naturales Renovables
Ing. Edgar Benítez
E-mail: agropecuaria@unl.edu.ec
Pontificia Universidad Católica del Ecuador
Facultad de Ciencias Exactas y Naturales
ESCUELA DE CIENCIAS BIOLÓGICAS
Alvaro Barragán
E-mail: arbarragan@puce.edu.ec (Interesado)
E-mail: webmaster@puce.edu.ec (Devuelto)

BRASIL
Dr. José Willibaldo Thomé
Facultade de Biociéncias
PUCRS. Av Ipiranga, 6681
Predio 12-D. 90.619.900
Porto Alegre, Río Grande do Sul
Brazil
E-mail: thomejw@pucrs.br
Facultad Ciencias Biologicas: biociencias@pucrs.br
Lenita de Freitas Tallarico (Biologist)
Laboratorio de Parasitología/Malacología
Instituto Butantan
Avda. Voital Brasil, 1500
CEP-05503-900 Sao Paulo. Brasil
E-mail: letallarico2@butantan.gov.br
Facultad de Engenheria Agricola
Brazil
Faculdade de Engenharia Agrícola
Campus Universitário, s/n
96010-900 Capão do Leão, RS
Fone: (53)32757317
Fax: (53) 32757373
E-mail: jluiz@ufpel.tche.br (Devuelto)
MARIA LAURA GOMES SILVA DA LUZ
Processamento de Produtos Agropecuários
E-mail: lauraluz@terra.com.br

COLOMBIA
Dra. Marleni Ramírez
Directora Regional de Biodiversity Internacional
CGIAR Bogotá, Colombia
Correo-e: m.ramirez@cgiar.org
----------------------------------------
Dr. Luis Enrique Vega
CONIF Director ejecutivos
Bogotá, Colombia
Correo-e: enriquevega@conif.org.co
------------------------------------------------
Dra. Clara Inés Medina Bermúdez
Universidad Militar de Nueva Granada
Bogotá. Colombia.
E-mail: cmedinabster@gmail.com
clara.medina@unimilitar.edu.co
------------------------------------------------------
Dra. Luz Elena Velásquez Trujillo
Coordinadora Línea de Investigación Malacología Médica & Tremátodos
Programa de Estudio y Control de Enfermedades Tropicales –PECET
Universidad de Antioquia
Medellín, Colombia
E-mail: luzelena333@yahoo.com
-----------------------------------------------------
Dra. Berta Miriam Gaviria G.,
Dr. Rafael Navarro Álzate
Laboratorio de Sanidad Vegetal
Universidad Católica de Oriente
Rionegro. Medellín
Colombia. Teléfono: 316666 Ext. 299
E-mail: rnavarro@uco.edu.co
E-mail: bgaviria@uco.edu.co
------------------------------------------
Facultad de Ciencias Agropecuarias
Universidad Nacional de Colombia, Sede Medellín
Calle 59A No. 63-020, Bloque 41, Oficina 209
Teléfono: (574) 430 90 00 - Fax: (574) 230 04 20
Colombia, Medellín, Antioquia.
Departamento de Ciencias Agronómicas
Alberto Álvarez Cardona
E-mail: posgeamb@unalmed.edu.co (Devuelto)
reuna@unalmed.edu.co
apcastilo@unalmed.edu.co (Devuelto)
fcaviaca@unalmed.edu.co
oriunmed@unalmed.edu.co
gcorrea@unalmed.edu.co
-------------------------------------------------------
Facultad de Agronomía en Bogotá
Universidad Nacional de Colombia
Carrera 45 No 26-85 - Edificio 500
Bogotá D.C. - Colombia
Prof. Augusto Ramírez Godo
Correo Electrónico: augramirezg@unal.edu.co
-------------------------------------------

CUBA
Universidad de la Habana:
Dra. Alicia Otazo
Decano Fac. Biologia. Cuba
E-mail: aotazo@fbio.uh.cu
---------------------------------------
Lic. Carlos Manuel Pérez Cueva
Vicerrector. Cuba
E-mail: carlosm@rect.uh.cu
------------------------------------------
Dr.Gustavo Cobreiro Suárez
Rector Universidad. Cuba
E-mail: rector@rect.uh.cu
-----------------------------------
Dr. Alejandro Barro
Facultad de Biología
Departamento de Biología Animal y Humana
Universidad de La Habana,
Calle 25, No. 455, Vedado,
Ciudad de La Habana, Cuba.
E-mail: abarro@fbio.uh.cu

COSTA RICA
Dr. Galileo Rivas (Contacto)
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
Tel: + 506 25582391
Fax: + 506 25582045
E-mail: grivas@catie.ac.cr
www.catie.ac.cr
---------------------------------------------
John Beer
CATIE
Costa Rica
Correo-e: jbeer@catie.ac.cr
-----------------
José Joaquin Campos Arce
Director General CATIE
Costa Rica
Correo-e: dbarquer@catie.ac.cr
--------------------
Francisco Javier Enciso Duran
Especialista Regional en Tecnología e Innovación IICA
Costa Rica
Correo-e: Laura.Mendez@iica.int
------------------------
Nicolás Mateo Valverde
Secretario ejecutivo. FONTAGRO
Costa Rica
Correo-e: fontagro@iadb.org
------------------------------------------

MEXICO
Dra. Edna Naranjo-García
Departamento de Zoología
Instituto de Biología
Universidad Nacional Autónoma de México
Apartado Postal 70-153
04510 México, D.F.
México
E-mail: naranjo@servidor.unam.mx
Facultad de Biologia
Edificio "R". Ciudad Universitaria.
Av. Fco. J. Múgica s/n. Col. Felícitas del Río
Tel. y Fax (443) 3.16.74.12 C. P. 58030
Morelia, Michoacán, México
E-mail: biologia@jupiter.umich.mx

PARAGUAY
Universidad Nacional de Asunción
Dr. LUIS GUILLERMO MALDONADO CHAMORRO
Director de Ingeniería Agronómica
FACULTAD DE CIENCIAS AGRARIAS (CAMPUS UNIVERSITARIO) SAN LORENZO PARAGUAY
E-mail: dircia@agr.una.py
Claudia Carolina Cabral Antunez. M.Sc. - Ing.Agr.
Directora del Departamento de Protección Vegetal.
Asignaturas: Manejo Integrado de Plagas, Control Biológicos de Plagas, Entomología I
Alicia Susana Aquino Jara. M.Sc. - Ing.Agr.
Jefa de la División de Fitopatología
E-mail: protvege@agr.una.py
Asignaturas: Microbiología, Fitopatología
Percy Antonio Salas Pino, M.Sc., Ing.Agr.
Jefe de la División de Malezas
E-mail: julios98@yahoo.com.
Asignaturas: Agronomía General, Cultivos I, Malezas II
Darío César Pino Quintana, M.Sc., Ing.Agr.
División de Fitopatología
E-mail: dariopino1@hotmail.com
Asignaturas: Agronomía General, Microbiología,Protección Vegetal I
Cristhian Grabowski, Ing.Agr
División de Fitopatología.
E-mail: cjgraboswski@yahoo.com.
Asignaturas: Jefe de Trabajos Prácticos de Microbiología, Fitopatología y Patología Forestal.
Maria Bernarda Ramirez de Lopez Ing. Agr.
Divsión Entomología.
E-mail: mabramirez@hotmail.com
Marcos Salvador Villalba Vásquez
Paraguay
Correo-e: dia@telesurf.com.py (Devuelto)

VENEZUELA
Dr. Orlando Moreno
Gerente General INIA
Venezuela
Correo-e: yreyes@inia.gob.ve
-------------------------------------
Director: Camarada Hernán Nieto.
e-mail: hnieto@inia.gob.ve
Dirección: Av. Urdaneta. Edif. INIA al lado del MAT.
Mérida, estado Mérida.
Benezuela
email: me_dir@inia.gob.ve
-----------------------------------------
Universidad Central de Venezuela
Facultad de Agronomía
Delia Polanco: polanco.delia@yahoo.es
M.A. Hortalizas Mauro Albarracin E-mail: albarracinm@agr.ucv.ve
Universidad Central de Venezuela
Facultad de Agronomía
Decano: Dr. Leonardo Taylhardat
E-mail: taylhardatl@agr.ucv.ve (Devuelto)
Coordinadora de Estaciones Experimentales
Prof. Xiomara Abreu
E-mail: abreux@agr.ucv.ve
IBE Instituto de Biología Experimental
Dirección: Calle Suapure, Colinas de Bello Monte, Caracas Venezuela.
Apartado postal: 47114, Caracas 1041A, Venezuela.
Teléfonos: (58 212) 751-0111
Fax: (58 212) 753-5897 (Devuelto)
E-mail: diribe@ciens.ucv.ve
Universidad Simón Bolívar
Decanato de Investigación Sartenejas
Edificio de Mecánica y Materiales, Piso 3.
Teléfono: 58-0212-9063900/9063901/9063902
Fax:58-0212-9063903
Correo electrónico: dec-id@usb.ve Devuelto)
Página web: http://www.did.usb.ve/
Dirección de Investigación Sede Litoral.
Edificio de Mecánica y Materiales, Piso 3.
Teléfono: 58-0212-9063900/9063901/9063902
Fax:58-0212-9063903
Correo Electrónico: div-inv@usb.ve
Página Web: http://www.dir-inv.nul.usb.ve/