Ziel
GENERAL DESCRIPTION OF THE ACTION
A. INTRODUCTION
Entomopathogenic nematodes of the genera Steinernema and Heterorhabditis are associated with bacteria of the genus Xenorhabdus. The infective nematode dauer juvenile, adapted for long term survival in the soil, carries cells of its symbiont Xenorhabdus sp. in the intestine. It actively seeks suitable host insects, penetrates and releases the bacteria. Providing the insect's defence mechanisms do not succeed in eliminating the nematode-bacterium complex, the insect dies two days after infection. The bacteria proliferate, the nematodes feed on bacteria and insect tissue and propagate within the dead host. After two weeks the dauer juveniles leave the cadaver searching for new target insects in the soil.
Nematodes possess many attributes of an ideal biological control agent: they are environmentally safe, can be produced in large-scale bioreactors, are easily applied using standard spraying equipment and are compatible with most chemical pesticides. Three species are currently used in integrated pest management and approximately 300ha were treated with nematodes in 1992 in Europe. Due to high production costs and inconsistent control results in the field, the application is still limited to high value crops (mainly ornamentals and home gardens).
With Council Directive 91/414/EEC of 15 July 1991 concerning the placing of plant protection products on the market (1) becoming national law in Member States in August 1993, the use of several insecticides will be banned. In particular, those used for insect control in soil environments will no longer be available. The pesticide market potential of several high-value crops (ornamentals, fruit and vegetables) is limited, and industry will avoid costs related with registration of new insecticides for these cultures. Nematodes do not need to be registered and therefore represent an environmentally safe alternative to fill the gap. However, this development has to be accompanied by intensive R&D efforts to improve the efficiency of entomopathogenic nematodes together with production and formulation techniques.
B. GENERAL BACKGROUND
The main objective of the Action is the search for, and selection of, highly active nematode and bacteria strains in order to guarantee consistent control results. Research on related fundamental and applied problems, like identification, in vitro liquid culture production, storage, genetic improvement of efficacy and storage-related traits and nematode ecology are also approached and all together contribute to an increasing use of entomopathogenic nematodes in integrated insect pest management.
The Action supersedes COST Action 812 "Cold active lines of insect parasitic nematodes" which involved fourteen countries, and over all 28 laboratories. Since 1992, two Eastern European countries (Croatia and Hungary) have joined Action 812 and a Hungarian laboratory has been awarded a PECO grant to participate in the Action. The Action will allow a further intensification of the scientific collaboration between European research institutes, plant protection services and industry. Several enterprises started nematode production during the period of Action 812. In awareness of the increasing research activities, an international agro-biotech company recently transferred parts of their R&D activities to Europe and, in the future, intends to participate in the Action. Similar networks exist in the USA and Canada. The Action would be the only co-operation scheme with Europe.
C. OBJECTIVES OF THE ACTION
The main objective of the Action is to combine interrelated European expertise in order to increase the use of entomopathogenic nematodes in integrated pest management and further to reduce chemical control. In order to guarantee consistent out-door control results, further fundamental and applied R&D is needed. Many research fields are shared by different participating laboratories. To improve co-ordination of research fields and objectives and achieve a maximum co-operation within Europe, different working groups shall be initiated. The objectives of these working groups are given in chapter D.
D. SCIENTIFIC PROGRAMME
It is necessary to improve the structure of the collaborative work. Therefore, the following working groups will be established.
Working group 1: Isolation and identification
Many new nematode strains were isolated during the period of Action 812 and participants were trained in methods for identification of species during a workshop in 1992. The objective of this working group is further to improve methods for isolation, morphological and molecular identification and cryopreservation of entomopathogenic nematodes. Participants of the new Action will support the setting up of a European strain collection at the CAB (Commonwealth Agriculture Bureau, London, UK) with services for identification and possibilities for the safe depositing of strains. At the same place a data-base will be established making available information on species and strain-related characters. The data-base will also help to determine the distribution of different species in Europe. Data-base and strain collection will be a public service organized by the co-ordinator of this working group.
Working group 2: Production and application
During Action 812 participants organized a workshop on methods to evaluate the control potential of entomopathogenic nematodes. Several cold active strains have been identified. Some strains have already been successfully tested against larvae of the Black Vine Weevil, a pest in ornamentals and strawberries.
In order to utilize the overall control potential of entomopathogenic nematodes in integrated pest management, new target host shall be screened for susceptibility. At the same time, test systems to evaluate "beneficial" traits will be further developed and used for selection of new species and strains with improved performance under glasshouse and outdoor environmental conditions. Co-operation in order to elaborate improved evaluation methods for field trials will be intensified. Only three nematode species are currently commercially available. However, other species and strains with higher efficacy against certain target insects have already been selected. The co-operation on production will help make available necessary quantities of these strains for field testing. Formulation techniques shall be improved to prolong nematode shelf life and to improve their activity. Methods for quality control shall be improved with, as final objective, agreement on standard control procedures. The effect of agrochemicals on the performance of entomopathogenic nematodes shall be further tested.
Working group 3: Nematode ecology
Nematode persistence and population dynamics are poorly understood. In the past, entomopathogenic nematodes have been used only for inundative control. The potential of long-term establishment of selected and/or improved strains has so far not been tested. Information on the host specificity and the effect on non-target hosts is a prerequisite for inoculative release of exotic strains into agro-environments.
Working group 4: Xenorhabdus
The significance of the bacterial symbiont for nematode performance at low temperature and particularly the nutritional relation during nematode propagation has gained major importance in the last few years. Typical for all known Xenorhabdus species is an instability of the wild type. The so-called primary variant undergoes certain morphological and physiological changes leading to a secondary variant. This phase-shift occurs under in vitro conditions and coincides with unknown lethal effects to the nematodes during propagation. Little is known of the physiological and genetic mechanisms inducing this phase-shift. For an improvement of biotechnical mass-production, joint research activities on the bacterial symbionts are necessary and will concentrate on bacterial pathogenicity mechanisms and activities the better to understand the physiological and genetical background of the phase-shift.
Working group 5: Nematode biology and genetics
Entomopathogenic nematodes are excellent targets for genetic improvement. Their close relation to the model nematode Caenorhabditis elegans make possible the use of expertise and methods applied in animal physiology and genetics. Within Action 812 a workshop on nematode genetics was organized. To profit from the large amount of scientific knowledge gained in C. elegans research, further understanding of the biology and physiology of entomopathogenic nematodes has to be obtained. Metagenesis shall be used to isolate marker mutants for characterization of the genetic background of "beneficial" traits. Inbred lines will be used to calculate heritability of traits. Cross breeding of characterized lines shall be applied to create nematode strains with improved field performance and liquid culture production potential. This research field is currently approached by several scientific groups willing to participate in the new Action. Selective breeding will be applied in order to increase field efficacy and stability under storage conditions.
E. TIMETABLE
The Action is planned for five years. It is anticipated that the research outlined in section D will be carried out in parallel. Progress and final achievement of the objectives will depend greatly on the activities within the national scientific groups participating. The Action will help to accelerate the progress by co-ordinating the research objectives and joint activities.
F. ORGANIZATION, MANAGEMENT AND RESPONSIBILITIES
The co-ordination of the Action will be carried out by the Management Committee in accordance with Annex I to the Memorandum of Understanding. The following activities will be realized:
The Management Committee will assign working group co-ordinators. Co-ordinators will organize the co-operation within and between the working groups, organize sessions on topics within the objectives of their working group during COST workshops. The objectives and organization of the workshops will be proposed by the co-ordinators and discussed in meetings of the Management Committee. The editing of reports and joint publications on the results of the co-operative research will be in the hands of the co-ordinators.
The joint exploitation of the research potential shared by the participants will be realized by exchange of experts, postdocs and post-graduates between the laboratories financed from available national and international sources and by exchange of information through workshops. Each year at least one workshop will be organized. These workshops will be a forum for the distribution of results and improved or standardized protocols and techniques. The workshops shall not be a substitute for scientific meetings. The Management Committee will try to channel the presentation of results obtained within the collaborative work to scientific symposia or meetings of existing scientific organizations.
G. ECONOMIC DIMENSION
The annual economic dimension of the Action was estimated on calculations from national co-ordinators of all participating countries. The estimate was based on personal costs and national and international research funds available in 1993:
Scientific personal : 70 man-years
Technical personal : 25 man-years
Research funds : ECU 1,8 million/year.
Several achievements have been obtained so far by COST 819.
The very confusing taxonomy of the nematodes has been clarified by WG1. The International Institute of Parasitology (IIP) at St Albans (UK) is centralising a database for this purpose to identify new isolations of EPNs on the basis of the morphology compared with the appropriate molecular probes (see meetings section). Similarly, WG4 is standardising bacteriological methods for the taxonomy of the symbionts in order to promote a parallel international database for the symbionts of EPNs. These two important points have to be well established to define accurately such biological material for industrial purposes, specially when exotic nematodes are planned to be used in Europe. On the other hand, COST 819 will contribute to international efforts on the studies of the biodiversity.
This is the reason that WG2 organised a workshop in Malente (D) on 8/11 June 1995 in order to define the risks for the human and animal health, and for the environment (see meetings section). No risks were identified and some guidelines were recommended (see publications).
During the WG2 meeting (held at Gosier on 12 July 1996) on the use of nematodes in tropical areas, the network of entomopathogenic nematodes in the Caribbean basin created and organised by the INRA laboratory in Guadeloupe (F) including 7 different countries (Cuba, The Dominican Republic, Guadeloupe and depending islands, Martinique, St Vincent, Trinidad & Tobago, Jamaica and Puerto Rico) was described by N. Boemare as an international extension of the COST 819. Similarly the IIP in St Albans (UK) is collecting samples from East Africa and South Asia. These networks will provide systematic and geographical data on the world distribution of EPNs.
Two important inventories are being checked : "Natural occurrence of EPNs in soil" gathered by J. Coosemans (WG2), and "Natural occurring infections of insects with EPNs" gathered by A. Peters (WG3). These surveys will be concluded soon.
A home page describing activities of the COST 819 is ready to be incorporated in the EU Internet facilities in early-1997.
Several companies are producing or selling nematodes in the EU. The list of the products and the companies will be available on theWeb. The most recent firm was E-Nema created by our previous Chairman, R-U Ehlers, in connection with the University of Kiel.
Current status
The MoU of the COST Action 819 was signed in Brussels on 24 May 1994 and by the end of 1996 17 European countries and one Research Institute in Israel had already signed it. The inaugural meeting of the Management Committee (MC) took place in Brussels on 6/7 July 1994. The 7th MC held in Ponta Delgada (P) on 21 March 1996 elected unanimously N. Boemare (F) as Chairman and R-U. Ehlers (D) as Vice-Chairman.
Working Groups
Five Working Groups have been set up; at the 8th MC, held in Gosier (Guadeloupe, F) on 12 July 1996, and during the 9th MC held in Leuven on 13 December 1996 there were changes in those appointed to convene the Groups. Working Groups and present Convenors are as follows :
Working Group 1 (WG1), "Isolation and Identification" : B. Hominick (UK) and A. Burnell (IRL)
Working Group 2 (WG2), "Production and Application" : J. Coosemans (B) and J. Grüner (CH)
Working Group 3 (WG3), "Nematode Ecology" : C. Griffin (IRL) and P. Smits (NL)
Working Group 4 (WG4), "Symbionts and Pathogenicity mechanisms " : N. Boemare (F) and N. Simoes (P)
Working Group 5 (WG5), "Nematode Biology and Genetics" : A. Burnell (IRL) and A. Fodor (H)
Short term scientific missions
Many Short Term Scientific Missions (STSM) have been carried out, strengthening interactions between our laboratories in order to consolidate collective knowledge from the specific expertise of each laboratory and to acquire new experimental tools for improving nematodes as biological control agents. Some recent examples : A. Peters (I) went to Wageningen Agricultural University (NL) (15 Jan/15 Feb 1996) for training courses on modelling to simulate systems management in crop protection with entomopathogenic nematodes. P. Fitters (IRL) trained in the Research Station for nursery Stock (NL) (15/21 Jul 1996) to develop the control of the black vine weevil (Otiorrhynchus sulcatus) in orchards and nurseries. Z. Kiss (H) attended a training course at Max Planck Institute (D) (1/30 June 1996) to study genetics of the pigment operon in Photorhabdus. C. Ribeiro (P) carried out a project in the Laboratory of Comparative Pathology (F) (1/30 Sep 1996) to purify by chromatography the toxic proteins produced by helminthic bacterium complexes and to study their effects on cellular systems by electron microscopy. M. Nugent (IRL) visited Wageningen (NL) (15 Jul/12 Aug 1996) to improve the cryopreservation of the nematode stocks of the P. Smits laboratory and to learn about the rearing of soil insects. Following the first isolation of 4 satellite DNA probes, recently promoted by the INRA Laboratory of Nematology in Antibes (F) to characterise Steinernema carpocapsae, Steinernema glaseri, Heterorhabditis bacteriophora and Heterorhabditis indicus, L. Waeyenberge (B) attempted to isolate satellite DNAs from Steinernema affinis (18/29 Nov 1996); experiments started in France are now continuing in Belgium.
Wissenschaftliches Gebiet (EuroSciVoc)
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CORDIS klassifiziert Projekte mit EuroSciVoc, einer mehrsprachigen Taxonomie der Wissenschaftsbereiche, durch einen halbautomatischen Prozess, der auf Verfahren der Verarbeitung natürlicher Sprache beruht. Siehe: Das European Science Vocabulary.
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