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COST 819


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Entomopathogenic nematodes: Application and persistence of entomopathogenic nematodes: Proceedings of a workshop held at Todi, Perugia, Italy, 16 to 20 May 1995
Agricultural and fisheries research



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in the field of
European Commission
scientific and
technical research
COST 819
Entomopathogenic nematodes
Application and persistence
of entomopathogenic nematodes
25 ■
■Ό 20
Ι io
Edith CRESSON, Member of the Commission
responsible for research, innovation, education, training and youth
DG XII/B.1 — RTD actions: Cooperation with non-member countries and inter­
national organisations— European Economic Area, COST, Eureka and interna­
tional organisations
Contact: Ms F. Coudert
Address: European Commission, rue de la Loi 200 (SDME 1/39)
B-1049 Brussels — Tel. (32-2) 29 65507; fax (32-2) 29 64289
Cover page: The diagram indicates the number of entomopathogenic nematodes recovered from nema-
tode-treated plots by five groups of workshop participants. The field plots had been treated with nematodes
at intervals prior to the COST workshop. COST
°% European Commission in the field of
scientific and
technical research
COST 819
Entomopathogenic nematodes
Application and persistence
of entomopathogenic nematodes
Proceedings of a workshop
held at Todi, Perugia, Italy
16 to 20 May 1995
Edited by
R. L. Gwynn, P. H. Smits, C. Griffin,
R.-U. Ehlers, N. Boemare and J.-R Masson
Science, Research and Development
Neither the European Commission nor any person acting
on behalf of the Commission is responsible for the use which might be made of
the following information.
A great deal of additional information on the European Union is available on the Internet.
It can be accessed through the Europa server (http://europa.eu.int).
Cataloguing data can be found at the end of this publication.
Luxembourg: Office for Official Publications of the European Communities, 1999
ISBN 92-828-6887-7
© European Communities, 1999
Reproduction is authorised provided the source is acknowledged.
Printed iâ Belgium CONTENTS
Some considerations on the application of entomopathogenic nematodes into field
crops, by Andrew C. Chappie 9
Entomopathogenic nematode application - a practical demonstration, by Andrew C.
Chappie and Roma L. Gwynn 21
Key factors affecting the commercial development of entomopathogenic nematode
products, by Sara Collins5
Post-application incorporation of Phasmarhabditis hermaphrodite into soil for slug
control, by Michael Wilson, Louise Hughes, Maria Hamacher, Dejanira Barahona
and David Glen9
Application of entomopathogenic nematodes on plant foliage, by Itamar Glazen... 37
Nematode extraction, by Jan van Bezooijen 61
Differentiating entomopathogenic nematodes from other living nematodes, by Arne
Peters 73
Persistence of entomopathogenic nematodes, by Peter H. Smits 77
Post-application persistence of entomopathogenic nematodes - A practical
demonstration, by Roma L. Gwynn, Andrew C. Chappie and Peter H. Smits 89
Early death of entomopathogenic nematode infective juveniles : the unsolved
question, by Birgit Hass 95
Nematode persistence in the presence of hosts : interpretation of field results with
Heterorhabditis spp. against two grub species in golf course turf, by Didik
Sulistyanto, Giulio Marini and Ralf-Udo Ehlers 101
Influence of host and plant roots on the migration of Heterorhabditis sp. (NWE) in
peat soil, by Robert W.H.M. van Tol and Ammy C. Schepman 117 NATURAL OCCURRENCE
Prevalence and habitat specificity of entomopathogenic nematodes in Germany, by
Dieter Sturhan 123
Incidence of the entomogenous families Steinernematidae and Heterorhabditidae
(Nematoda) and their dependence on insect outbreaks, by Zdenek Mrácek 13
Distribution of entomopathogenic nematodes in grassland habitat, by John S.
Miduturi, Maurice Moens, and R. Moermans 141
Survey of the research about « The natural occurrence of entomopathogenic
nematodes in the soil », by Ruth Stoffelen and Josef Coosemans 145
Surface appendages of phase one and two forms of the nematode-associated
bacterium Xenorhabdus nematophilus, by W. Li and Martin Burman 169 FOREWORD
The use of nematodes for the biological control of soil-borne pests has gained
increasing interest in the past few years. Coleopteran and dipteran insect larvae and
slugs are nowadays successfully controlled with nematode-based products, mainly in
glasshouse crops. The challenging objective for future R&D activities is to provide
basic research results which can enhance the large scale outdoor use of these
antagonists. Nematodes can only reveal their full control potential if they are
successfully transferred to the soil environment and survive at high population
densities until a reasonable control result has be achieved. Apart from the solution
of constrains related to the commercialisation of nematode products, nematode
application and persistence was identified as a major problem by the community of
researchers active within COST Action 819 "Entomopathogenic Nematodes".
Therefore a workshop was organised to introduce and discuss latest progress made
in related fields.
COST (Co-operation in Science and Technology) is a programme of the Commission
of the European Union (DG XII). The Commission deserves our acknowledgement
for the financial support of this workshop. COST Action 819 "Entomopathogenic
Nematodes" represents a forum for science and industry to co-ordinate their efforts
and enhance scientific and technical co-operation within Europe. This workshop
was held at the facilities of a commercial company and it was organised by Roma
Gwynn and her colleagues at Ecogen Europe S.r.l., Todi-Perugia, Italy. On behalf of
all COST 819 participants I want to thank the Ecogen team for the excellent
organisation which made us enjoy the science and the nice surrounding in the
Umbrian region. Thanks are also due to Bruce Carlton (Ecogen Inc.) and Dorio Mutti
(3A S.r.l.), who supported my efforts to continue with our plans to hold the meeting at
the Todi facilities. Thanks are also due to all the scientists presenting their results
and ideas at the workshop. Particular gratitude deserve all colleagues who made
possible the practical sessions on nematode application and recovery from soil
Due to technical problems the publication of these proceedings had to be delayed,
however, their content has by no means lost its value for the discussion on nematode
performance in the soil environment and will influence the future research directed at
a better understanding of nematode performance in the soil and how these biocontrol
agents can best be applied.
Raisdorf, August 14, 1996
Ralf-Udo Ehlers
Chairman COST 819 I have to emphasise the comments of the previous chairman. All the contents of
these proceedings are always useful. So we decided to publish these proceedings
which have been gathered after a long period of uncertainty. Many thanks to Roma
Gwynn for her efforts.
Montpellier, December 11, 1998
Noël Boemare
Amaral Joao S.D.A.T., Vinha Brava, P­9700 Angra do Heroísmo, Açores,
Andermatt Martin Andermatt­Biocontrol Ag., Unterdorf, CH­6146 Grossdietwil
Institut für biologische, Schädlingsbekämpfung, Biologische Bundesanstalt, Bathon Horst
Heinrichstr. 243, D­64287 Darmstadt
Nematology Department,Wageningen Agricultural University, Binnenhaven, van Beezoijen Jan
10, NL­6709 PD Wageningen
Bocelli Mauro Osservatorio per le malattie delle piante, Via di Coticella, 133, 1­40129
Boemare Noel Lab.de Pathologie Comparée, INRA­CNRS URA 2209, CP101, Université de
Montpellier II, F­34095 Montpellier Cedex 5
Bonifassi Eliane Lab Biologie des Invertébrés, INRA, F­06606 Antibes Cedex
Burman Martin Dept. of Zoophysiology, University of Umea, S­90187 Umea
Carolli Luigi Ecogen Europe, Frazione Pantalla, I­06050 Todi(PG)
Chappie Andrew n , e Pantalla di Todi, I­06050 Todi (PG)
Collins Sara Agrisense, Unit 1, Taffs Mead Road, Trefforest Industriai Estate, Pontypridd,
Mid Glamorgan CF37 5SU, UK
Coosemans Josef Lab. Fytopathologie en Plantenbescherming, K U Leuven, Willem de
Croylaan 42, B­3001 Heverlee­Leuven
Curto Giovanna Osservatorio per le malattie delle piante, Via di Corticella, 133, 1­40129
Daoust Richard Ecogen Inc., 2005 Cabot Boulevard West, P.O. Box 3023, Langhome, PA
19047­3023, USA
Downes Martin Biology Dept., St Patrick's College, Maynooth, County Kildare, Ireland
Ehlers Ralf­Udo Inst, für Phytopathologie der Christian­Albrecht­Universität Kiel,
Arbeitsgruppe Biotechnologie, Klausdorferstr. 28­36, D­24223 Raisdorf
Francescini Sergio Ecogen Europe, Frazione Pantalla, I­06050 TODI (PG)
Gerber Karin Research Center for Agriculture, Trunnerstrasse 5, A­1020 Vienna
Gerritsen Lonne IPO­DLO, Binnenhaven 5, P.O.Box 9060, NL­6700 GW Wageningen
Glaser Itamar Dept. of Nematology, Volcani Center, P.O.Box 6, Bet Dagan 50250, Israel
Gottrup Ole Agrolab A/S, Røjleskovvej 18, DK­5500 Middelfart
Grewal Parwinder BIOSYS, 1057 East Meadow Circle, Palo Alto, CA 94303, USA
Griffin Christine Biol. Depart., St. Patrick's College, Maynooth, County Kildare, Ireland
Grunder Jürg Swiss Federal Research Station, Eidg. Forschungsanstalt, CH­8820
Gwynn Roma Ecogen Europe, Frazione Pantalla, I­06050 TODI (PG)
Hass Birgit Biol. Dept., St. Patrick's College, Maynooth, County Kildare, Ireland
Hominick Bill CAB International, Institute of Parasitology, 395A, Hatfield Road, St. Albans,
Hertfordshire AL4 OXU, UK
Jaworska Magdalena Inst. Plant Protection, Ave. 29 Listopada 48, 31­425 Krakow, Poland
Jung Kerstin B.B.A., Institute for Biological Control, Heinrich Str. 243, D­64287 Darmstadt
Klein Michael Horticultural Insects Res. Lab., OARDC, Wooster, OH 44691, USA .
Krasomil­Ost. Karina Ecogen BioGermany, Klausdorfer Str. 28­36, D­24223 Raisdorf
Long Steve Horticulture Research International, Entomology Dept., Worthing Road,
Littlehampton, West Sussex BN17 6LP, UK
Masson Jean­Pierre CCE , DG, XII/B/1, 200, rue de la Loi, Β 68 5/12­5/8, B­1049 Bruxelles
Miduturi John Rijksstation voor Nematologie en Entomologie, Van Gansberghelaan 96, B­
9820 Merelbeke
Moens Maurice Rijksstation voor e en , Van Gansberghelaan, 96, B­
9820 Merelbeke Mrácek Zdenek Institute of Entomology, Czech Academy of Sciences, Branisovska 31, 370
05 Ceske Budejovice, Czek Republic
Osterfeld Karl Ecogen Bio-Germany, Klausdorferstr. 28-36, D-24223 Raisdorf
Peters Arne Ecogen Europe, Frazione Pantalla, I-06050 Todi (PG)
Philipsen Holger Royal Veterin. & Agricult. University, Thorvaldoensveij 40, DK-1871
Frederiksberg, Copenhagen
Ragni Adriano Ecogen Europe S.r.l., Frazione Pantalla, l-06050Todi (PG)
Ricci Manuele n Europe, Frazione Pantalla, I-06050 Todi (PG)
Rick van der Pas Koppert B.V., P.O.Box 155, NL-2650 Ad Berkel & Rodenrys
Rob van Tol Research Station for Nursery Stock, P.O.Box 118, NL-2770 AC Boskoop
Rovesti Luciano Centra di Studio dei Fitofarmaci, Via Filippo Re, 8, 1-40126 Bologna
Simões Nelson Dept. de Biologia, Univ. dos Açores, Rua da Mae Deus, 58, P-9502 Ponta
Delgada, Açores
Smits Peter IPO-DLO, Binnenhaven, 12, Postbus 9060, NL-6700 GW Wageningen
Spiridonov Sergei Institute of Parasitology, Russian Academy of Science. Lenin Avenue, 33,
117071 Moscow, Russia
Sturhan Dieter BBA, Toppheideweg 88, D-48161 Munster
'Sulistyanto Didik Institut für Phytopathologie, Universität Kiel, Klausdorferstr. 28-36, D-24223
Taylor Robin Dept. of Entomology, OARDC, Ohio State University
Wooster, OH 44691, USA
Wegensteiner Rudolf Institute Forest Entomology, Hasenauerstr. 38, A-1190 Wien
Wilson Mike IACR, Long Ashton Research Station, Long Ashton, Bristol B518 9AF, UK