European Cooperation in the field of European Commission scientific and technical research COST 819 Entomopathogenic nematodes Application and persistence of entomopathogenic nematodes 30 25 ■ ■Ό 20 Î ε o i Ι io EUR 18873 EN EUROPEAN COMMISSION 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 cooperation °% 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 Directorate-General Science, Research and Development EUR 18873 EN 1999 LEGAL NOTICE 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 FOREWORD 5 LIST OF PARTICIPANTS 7 APPLICATION 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 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 BACTERIAL SYMBIONTS 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 samples. 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 2nd Chairman COST 819 LIST OF PARTICIPANTS 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 Bologna 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 Bologna 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 Wadenswil 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 Raisdorf 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