Practicalities of developing and registering microbial biological control agents

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1 Review Practicalities of developing and registering microbial biological control agents Sebastian Kiewnick Address: Agroscope Changins-Wädenswil, Research Station ACW, Plant Protection, Ecotoxicology and Soil Zoology, 8820 Wädenswil, Switzerland. Correspondence: Received: 29 November 2006 Accepted: 30 January 2007 doi: /PAVSNNR The electronic version of this article is the definitive one. It is located here: g CABI Publishing 2007 (Online ISSN ) Abstract CAB Reviews: Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources , No. 013 There is considerable interest in the exploitation of microbial biological control agents (MBCAs) for the control of crop pests, weeds and diseases. MBCAs can be used where chemical pesticides are banned or being phased out or where pests have developed resistance to standard chemicals. The use of MBCAs can play an important role in crop protection, as a key element in integrated pest management (IPM) programmes. However, despite considerable research efforts on the development of new biological control agents the number of such products on the market in the European Union (EU) is still extremely low compared with the United States or Canada. In areas that previously constrained the commercialization of MBCAs, discovery, fermentation, formulation and application, significant progress has been made. The low number of products is mainly due to the slow registration process. In the EU, MBCAs are regulated by and follow Directive 91/414/EEC for placing plant protection products in the market. Once an active ingredient is listed in Annex I, national registrations for the formulated product have to follow. This time consuming and expensive process has forced most companies to suspend their efforts in research and development. Initiatives by stakeholders from industry, science, regulatory authorities, policy and environment are underway to accelerate market introduction of MBCAs. Keywords: Biological control, Production, Fermentation, Formulation, Registration Review Methodology: The following databases were searched: CAB Abstracts and Pubmed (Keyword search terms used: registration, biological, control, agent, registration, microbial, biopesticide). References from the articles obtained by this method were used for additional relevant material. In addition, information from a symposium at the APS, CPS, MSA joint meeting July 29 August 2, 2006 in Que bec City, Que bec, Canada entitled Research, Development, and Adoption of Biopesticides in the 21st Century and a workshop on Current Risk Assessment and Regulation Practice September 2006 in Salzau, Germany with more than 100 experts (scientists from public research and industry, and government regulators) were included. Introduction Biopesticides, by the general definition, are pest limiting agents of biological origin which include microbial living systems (bacteria, fungi, viruses), entomopathogenic nematodes, insect predators and natural parasites, plant derived products (botanicals) and insect pheromones (natural and semiochemicals) [1, 2]. Under United States Environmental Protection Agency (US EPA) regulation, the use of plant incorporated protectants (such as Bacillus thuringiensis (Bt)-toxins in transgenic plants), genetically engineered micro-organisms, biochemical compounds with a non-toxic mode of action and biochemical like compounds appropriate for reduced data are also considered biopesticides [3]. Given the large number of biopesticide products [4], this review can cover only a fraction of this large group and refers to microbial pesticides or microbial biological control agents (MBCAs). Products containing bacteria, fungi or viruses are currently receiving a lot of attention from researchers, industry and authorities [2, 5, 6]. In 1996, the predicted market size for biocontrol products for the year 2000 was 10% of all pesticides sold around the world, with a value of $3 billion [7]. Today, the

2 2 Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources world pesticide market is estimated at $25 billion, but only 1% ($300 million) is spent on biopesticides. In the European Union (EU), the total pesticide market is estimated at $5 billion with 2% of this total volume being biopesticides ($100 million). Of this $100 million only 25% comes from sale of MBCAs. The largest proportion of these MBCAs are Bt products with 80 90% of the market and only $2.5 5 million comes from non Bt-sales [2]. Despite the fact that scientists have been working for over 50 years on biological control and integrated pest management (IPM) systems, the biocontrol business is growing a rate of only 10% per year [8]. Consequently, only a few decent medium-sized companies make a profit selling biological products. Most biopesticide companies are unprofitable or marginally profitable [9]. Perceptions of MBCAs There are three major unique features of MBCAs which make them of interest to a farmer: (i) resistance management (ii) restricted entry intervals and (iii) residues. Because most MBCAs have multiple modes of action [10, 11] there is less chance of resistance developing in a particular pathogen, insect or weed. Therefore, MBCAs are an excellent tool in IPM programmes, where synergistic effects can be utilized to reduce the input of chemicals or to re-establish efficacy [6, 12]. The majority of MBCAs if not all have low restricted entry intervals, normally in the range of 0 4 h [9]. In addition, they generally have no pre-harvest interval, which essentially allows the farmer to harvest a crop directly after an MBCA has been applied. Finally, MBCAs are generally considered exempt from tolerances (maximum residue limits) under US EPA regulation. The US EPA considers these products safe and therefore that residues on leaves and fruit do not pose an unacceptable risk [9]. Under EU regulation, residue data are not relevant only if no adverse effects are identified from the proposed use. Residues are here considered as toxins produced by micro-organisms that may be of biological significance. If produced by a micro-organism, maximum residue levels (MRLs) have to be established for these toxins. MCBAs in the EU, US and Canada Despite progress made in research and development associated with the use of MBCAs, the use of these products in the EU is still very limited. As demonstrated in Tables 1a, b, c and 2, the number of products registered in the EU in comparison with other countries (e.g. the US) is very small. Tables 1a, b and c list 68 MCBAs used as bactericides, fungicides, herbicides, insecticides and nematicides that fulfill at least one of the following requirements: (1) registration as an active substance in the US; (2) registration as an active substance in Canada; (3) registration as an active substance in an Organization for Economic Co-operation and Development (OECD) member state outside the EU; (4) registration in an EU member state as an old active substance or with a provisional registration; (5) inclusion of the active substance in Annex I of Directive 91/414/EEC; (6) under evaluation as a list 4 substance as either a new existing or an old active substance. In order to limit the list to a reasonable length Bt and insect virus products are not listed in detail. The Bt group alone represents 13 active ingredients in 106 products on the US market only [17]. A detailed list of products containing Bt or viruses as active substances can be found elsewhere [18, 19]. A total of 53 MBCAs is currently registered in the US. In Canada and other OECD countries 15 and 20 MBCAs are registered, respectively. For the EU, 21 MBCAs are listed that have a registration in a least one member state, either provisional or as old active substances. Only five MBCAs are currently listed on Annex I of Directive 91/414/EEC, with four more active substances under evaluation. Concerning the last stage of the EU-evaluation process, the fourth list, 18 MBCAs have been notified with an additional three considered as new existing active substances. Besides the groups of Bt and virus products, only three other organisms, namely B. subtilis (Strain MBI 600), Streptomyces griseovirides (Strain K61) and Trichoderma harzianum have registrations in the US, Canada, other OECD and EU countries (Table 1a and b). The main reason for the difference in the number of products registered lies in the regulatory system that applies to microbial plant protection products in the EU. Regulation of MBCAs in the US, Canada and the EU The US regulatory requirements for microbial pest control products are outlined in Microbial Pesticide Data Tables 40 CFR established in However, US EPA recently proposed a change in data requirements for biochemical and microbial pesticides, taking into account that the current regulation was established 24 years ago and therefore data requirements needed adjustment [20]. In Canada, the Pest Management Regulatory Agency (PMRA) establishes the data requirements for microbial pest control products which are outlined in Regulatory Directive DIR In the EU, the placing of plant protection products on the market is regulated by Council Directive 91/414/EEC. Plant protection products containing micro-organisms as active substances are submitted to this directive, which has been modified by Commission Directive 2001/36/EEC to specify requirements for micro-organisms. The main differences between the regulatory systems in the US, Canada and EU are not within the overall data requirements. They are quite similar, but in the US and Canada, the description of the data requirements is more

3 Table 1a Comparison of the registrations of MBCAs bactericides and fungicides based on bacteria in the US, Canada, OECD countries and the EU Registration 1 EU (Directive 91/414 EEC) MBCA Target US Canada OECD 2 National 3 Annex I inclusion Notification List 4 Bactericides Agrobacterium radiobacter Strain K84 Crown gall New existing A. radiobacter Strain K1026 Crown gall Pantoea agglomerans Strain C9-1 Fireblight Pseudomonas fluorescens A506 Fireblight Fungicides (Bacteria) Bacillus cereus Strain UW85 Fungal soil-borne diseases B. licheniformis Strain SB3086 Fungal soil-borne and leaf diseases B. pumilus Strain GB34 Damping-off B. pumilus Strain 2808 Root and leaf fungal diseases B. subtilis Strain GBO3 Damping-off and soil-borne fungal diseases B. subtilis Strain IBE 711 Damping-off and soil-borne fungal diseases New existing B. subtilis Strain MBI 600 Soil and seedborne fungal diseases B. subtilis Strain QST713 Powdery mildew Pending N/A B. subtilis Damping-off B. subtilis subsp. amyloliquefaciens Soil-borne fungal diseases Strain FZB24 Paenibacillus polymyxa Strain AC-1 Damping-off, powdery mildew Pseudomonas aureofaciens Strain TX-1 Turf leaf diseases Pseudomonas chlororaphis Strain MA342 Cereal leaf diseases P. chlororaphis Strain Soil-borne diseases P. syringae Strains ESC-10, ESC-11 Fruit and dry rot Streptomyces griseovirides Strain K61 Damping-off and soil-borne fungal diseases Notified S. lydicus Strain WYCD108 Soil-borne fungal diseases Modified after Hynes and Boyetchko [13]. 1 Organisms are included that were listed in one of the following sources: [14 16]. 2 OECD countries other than US, Canada, and Europe. 3 Provisional registration or registration as old active substance in EU-member states (before 23 July, 1993). Sebastian Kiewnick 3

4 Table 1b Comparison between the registrations for MBCAs fungicides and herbicides based on fungi in the US, Canada, OECD countries and the EU Registration 1 EU (Directive 91/414 EEC) Annex I MBCA Target US Canada OECD 2 National 3 inclusion Fungicides (Fungi) Ampelomyces quisqualis Strain M10 Powdery mildew A. quisqualis Strain Powdery mildew Aspergillus flavus Strains AF36 and NRRL Aspergillus flavus Coniothyrium minitans Strain CON/M/91-08 Sclerotinia spp N/A Fusarium oxysporum Strain Fo47 Fusarium wilt diseases Gliocladium catenulatum Strain J1446 Damping-off and soil-borne N/A fungal diseases Notification List 4 G. virens Strain GL-21 Damping-off Muscodor albus Strain QST20799 Pre- and post-harvest diseases Ophiostoma piliferum Bluestain fungi Phlebiopsis gigantea Pine root rot Notified Pseudozyma flocculosa Strain PF-A22 Powdery mildew Pending N/A Pythium oligandrum Strain DV74 Soil-borne fungal diseases New existing Trichoderma harzianum, several strains Soil-borne and foliar fungal Notified diseases T. polysporum Pruning wound infection Notified T. viride Soil-borne fungal diseases Notified Verticillium dahliae Kleb. Dutch elm disease Notified Verticillium isolate WCS850 Dutch elm disease Notified Herbicides Alternaria destruans Strain 059 Cuscuta spp Chondrostereum purpureum Strain HQ1 Hardwood tree spp C. purpureum Strain PFC2139 Hardwood tree spp Colletotrichum gloeosporioides f. sp. aeschynomene Northern joint vetch C. gloeosporioides f. sp. malvae Round-leafed mallow Phytophthora palmivora Strain MWW Milkweed vine Puccinia thlaspeos Dyer s wood For footnotes (1 3) see Table 1a. 4 Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources

5 Table 1c Comparison between the registrations for MBCAs insecticides and nematicides in the US, Canada, OECD countries and the EU MBCA Target Registration 1 US Canada OECD 2 EU (Directive 91/414 EEC) Annex I National 3 inclusion Notification List 4 Insecticides (Bacteria) Bacillus thuringiensis (Berliner) Several insect species Notified B. thuringiensis subsp. aizawai Several insect species Notified B. thuringiensis subsp. israelensis Several insect species Notified B. thuringiensis subsp. kurstaki Several insect species Notified B. thuringiensis subsp. tenebrionis Several insect species Notified B. thuringiensis (engineered strain) Several insect species B. sphaericus Strain ATCC 1170 Mosquito larvae Notified Paenibacillus popilliae Japanese beetle Insecticides (Fungi) Beauveria bassiana, several strains Several insect species Notified B. bassiana Strain 447 Fire ants B. bassiana Strain HF23 House flies B. brongniartii Sugar cane beetle Notified Lagenidium giganteum Mosquito larvae Notified Metarhizium anisopliae var. acridium Strain Locust, Wingless IMI grasshopper M. anisopliae var. anisopliae Strain EFS-1 Termites M. anisopliae, several strains Several insect species Notified Paecilomyces fumosoroseus Apopka White fly N/A strain 97 P. fumosoroseus Strain FE9901 White fly Pending N/A Verticillum lecanii 4 (2 Strains) White fly and Aphids Notified Insecticides (Viruses) Granuloviruses, several strains Several arthropod species Notified Nucleopolyhedroviruses, several strains Several arthropod species Notified Nematicides Bacillus firmus Plant parasitic nematodes Paecilomyces lilacinus Strain 251 Plant parasitic nematodes Under evaluation N/A For footnotes (1 3) see Table 1a. 4 Taxonomic revision means the correct name for this genus is now: Lecanicilium. Sebastian Kiewnick 5

6 6 Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources Table 2 Comparison of the time needed (in months) for registering nine microbial biological control agents in the EU under Directive 91/414/EEC (i.e. inclusion in Annex I) compared with the US EPA Time needed for registration (months) Organism (strain) Product Europe (Annex I) US EPA Ampelomyces quisqualis (M10) AQ ? Bacillus subtilis (QST713) Serenade 1 > Coniothyrium minitans (CON/M/91-08) Contans Gliocladium catenulatum (J1446) Prestop Paecilomyces fumosoroseus (Apopka 97) Preferal Paecilomyces lilacinus (251) Bioact WG 1 > Pseudomonas chlororaphis (MA342) Cedomon 1 99 Pseudomyza flocculosa (PF-22) Sporodex 1 > Spodoptera NPV Spodex 1 > Average > Dossiers currently under evaluation;? = no data available. detailed and waivers (scientifically based arguments for non-submission of studies) are generally accepted. In addition, there is a guaranteed time line for the registration process [21]. Table 2 shows the time line for the registration of MBCAs by the US EPA compared with that under EU Directive 91/414/EEC for nine different biocontrol agents. On an average, the time needed for registration of a product in the US was 26 months (range 12 60), whereas more than 80 months (63 104) were needed for inclusion of an active substance in Annex I of Directive 91/414/EEC. During the years 1994 to 2003 more than 50 MBCAs were registered in the US, but in the EU only five active substances were added to Annex I. Furthermore, once the active substance has been included in Annex I, national registrations in EU member states of the formulated product still have to follow. As of August 2006, 78 microbial products were registered with the US EPA [3]. In the EU, eight new active substances are still under evaluation for inclusion in Annex I. Currently, stage four, which is the final stage of the review programme for the active substances already on the market in EU member states before 23 July 1993 is underway [22]. Dossiers for 140 of these old active substances have been submitted by notifiers, of which 19 dossiers are for microbial active substances (Tables 1a, b and c). With respect to the risk assessment, the evaluation in the US and Canada is based on a maximum hazard testing and not provided by the applicant. Conversely, in the EU, risk is evaluated on a hazardexposure basis and the applicant has to provide the risk assessment for the MBCA in the dossier submitted to the authorities. One of the major differences between the EU regulations and those of other countries lies in the requirement by the EU to provide efficacy data for the MBCA for all intended specific uses in specific crops [10]. Therefore, a product has to be evaluated in several experiments, in different geographic zones and for two consecutive years. As a consequence, products that have registrations for use on multiple crops against several diseases in the US will have a registration in the EU for a specific use only [10]. In the EU, this approach is used to prevent the application of products with no efficacy but potential unwanted side effects. Conversely, US EPA does not require efficacy data, because the attitude is that the market should decide whether a product is accepted by farmers or not. As a consequence, several products are currently registered with US EPA, but are not sold in significant quantities. Additional Reasons for the Low Number of Products Registered in the EU versus US and Canada As described above, the registration process is the main hurdle to overcome in the successful commercialization of an MBCA. There are some major problems within the regulatory process that are responsible for the delays in registration (Table 2). The guidelines currently used to evaluate MBCAs were originally developed for chemical pesticides and are mostly not appropriate for microorganisms [23]. In some cases (e.g. sensitization properties), authorities have recognized that methods for testing dermal sensitization are not suitable for testing microorganisms. Sensitization by inhalation is considered to be most probably a greater problem than dermal exposure, but no validated test methods are available [24]. As a consequence, all micro-organisms are regarded as potential sensitizers. This presumptive safety approach also takes into consideration immuno-compromised or other sensitive individuals in the population [24]. However, as a consequence, the formulated product carries an Xn-label which indicates that the content is classified as a sensitizer (R42/R43). From the standpoint of commercialization, this safety precaution has a tremendous impact on the handling, shipping and storage of a product that supposedly is a safe alternative to chemical pesticides. This definitely contradicts the general perception of biological control

7 Sebastian Kiewnick 7 (a) (b) c c National registrations of formulated product Figure 1 Basic procedures for the development and commercialization of a microbial biocontrol agent (a) general scheme modified after Montesinos [31] and (b) scheme demonstrating the requirements under current EU regulation products. In addition, products with an Xn-labelling cannot be stored longer with food in cold storage and the shipping and handling costs are significantly increased because a dangerous good is shipped. Consequently, the end user price increases, which makes an MBCA less attractive to the farmer and the product is no longer considered as safe by the end user [21]. The question whether toxins produced by microorganisms pose a risk to workers and consumers has fuelled the discussions on the safety of biocontrol agents in past years. Two EU funded Research, Technological Development (RTD)-projects, BIPESCO (Biological Control of Soil Dwelling Pests, FAIR6-CT ) and RAFBCA (Risk Assessment of Fungal Biocontrol Agents, QLK ), were initiated to enhance the production, formulation, and efficacy of fungal biocontrol agents and evaluate the risks involved with the application of these MBCAs [25]. One of the major concerns during the registration process of an MBCA, the production of metabolites, or toxins, was intensively investigated in the RAFBCA project. The objectives of the project were to identify and characterize metabolites produced by fungal BCAs, and to establish whether they entered the food chain and posed a risk to human and animal health based on new risk assessment tools. The major outcome of this research was that the potential for exposure to metabolites produced by certain fungal MPCAs was considered to be low [26 29] (for more details see www. rafbca.com). Changes in the Requirements for the Development of an MBCA The development of an MBCA requires several steps. However, the importance of each step and the impact of a single step on the whole process of discovering, developing and commercializing an MBCA has certainly changed over the last decade. This is especially true for EU member states and in part also for the US and Canada. Ten years ago, the strategies for selecting and developing microbial biopesticides for the control of insect weeds and plant pathogens focussed on technological constraints [30].These technological constraints included the lack of low-cost production methods, stable formulations with reasonable shelf-lives and efficacy under field conditions. Strategies for selecting potential biocontrol agents and experimental approaches to overcome the constraints were of key importance at that time. Figure 1 demonstrates how the general scheme of the steps important in the development of an MBCA has changed. This scheme, presented by Montesinos

8 8 Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources [31], placed emphasis on the initial steps of isolation, identification and characterization (Figure 1a). Until a few years ago, these first steps were rather laborious and resource intensive with a maximum success rate of 1% or less. Due to the advances in understanding the mechanisms involved and responsible for biocontrol activity, nowadays selection of potential biocontrol agents can be based on certain traits that are responsible for biocontrol efficacy [32]. Therefore, potential organisms can be selected at an early stage for further testing with an increased success rate [32]. Initial trials with the candidate strains are done under a range of conditions to fully evaluate their potential for disease control. In the new scheme (Figure 1b) this phase is of great importance for commercial and regulatory reasons [33]. At this point, the researcher should identify potential crops and cropping systems and determine effects such as physical, chemical or cultural factors that limit the effectiveness of the MBCA. Furthermore, in the context of regulatory issues, application timing and rates as well as pathogen thresholds have to be established early on in the development process. Once a candidate strain has been selected for further development, a decision on patenting has to be made. Patenting is particularly important for researchers who want to later licence their invention to a company. However, costs for obtaining and maintaining patents are high and although large number of patents on the use of MCBAs have been filed, only a fraction has materialized into registrations of the active substance [31]. This indicates that factors other than patents are more important for success in developing and commercializing MBCAs. Major improvements have been made in recent years in the process of mass production and formulation of microorganisms. The development and improvement of solid state fermentation technologies for filamentous fungi [34, 35] has led to the development of several new MBCAs. The advantages of solid state fermentation such as low capital investments and energy requirements combined with cheap and simple media have reduced the production costs to a level competitive with chemical pesticides [36, 37]. Furthermore, the development of new formulations of MBCAs have led to better storage stability, compatibility with standard application equipment and increased efficacy [13, 37]. The advances in the fermentation and formulation process have also led to an increase in the quality of biocontrol products. The possibility of contaminants (unwanted pathogens, toxins or toxic metabolites) in a biocontrol product needs to be considered at an early stage of development [38]. Field studies to evaluate the efficacy of the candidate organism must be conducted before an MBCA can be considered seriously for further development. Evaluation of product efficacy under diverse practical conditions can help to identify the limits of the MBCA. Furthermore, a compatibility profile of the MBCA with other crop protection inputs is essential for successful commercialization [39]. Lack of consistency in efficacy of biological products is still a problem [10]. However, a recent metadata analysis of efficacy data for MBCAs by Oijambo and Scherm [40] revealed that some of the previous assumptions concerning the efficacy of biocontrol products are not correct. Their study revealed a moderate effectiveness on average, but no differences between the effects of biocontrol agents in greenhouse versus field studies, between the effects on soil-borne versus aerial diseases, or under conditions of low, medium and high disease pressure. However, effects were greater on annual than on perennial crops, but were not different for fungal versus bacterial biocontrol agents or for those targeting fungal or bacterial pathogens. Interestingly, on average the efficacy of Bacillus spp. was lower than that of other antagonists. Finally, the effect of one or two sprays for control of aerial diseases was significantly greater than the application of eight sprays or more; this indicates that in an attempt to compensate for anticipated poor performance of biocontrol agents, more applications than necessary are made [40]. Recent developments in application technologies for MBCAs were reviewed by Gan-Mor and Matthews [41] who found several improvements in application technologies for biopesticides but concluded that yet more research and development is needed to make sure that promising products can be applied by farmers. Furthermore, advances in downstream processing and formulations of MBCAs, such as Bt and viruses, which have been in use for decades, allow further significant improvements in economy, shelf-life, ease of application and field efficacy [19, 42]. By improving production and formulation of these products even wider use can be expected in the future. In Figure 1a, the registration process is considered of equal importance to all other steps in the process of developing and commercializing an MBCA. Conversely, under the current EU regulatory system, several major hurdles have to be negotiated before the successful registration of a biocontrol agent (Figure 1b). Toxicological and environmental impacts are amongst the key issues in the current discussions concerning the registration process for MBCAs in the EU [25, 43, 44]. Better knowledge of the fate and behaviour of an introduced micro-organism in the environment is also crucial for appropriate assessment of potential side effects. It is desirable that the biocontrol agent is established in the area to which it is applied only for the period necessary for the control of the pathogen or pest in order to minimize risks that may ensue from its application. Longer persistence would increase exposure and the possibility of unwanted side effects [44]. MBCAs represent a complex array of approaches towards pest control. Their use for pest control presents challenges in understanding target selectivity, and the occurrence and fate of MBCAs in the environment [45]. Therefore, careful testing of human health risks

9 posed by the biocontrol agents is necessary [46]. However, experiences so far do not suggest an unreasonable risk of adverse health effects associated with MBCAs although the possibility of infectious and immunological responses needs consideration in the context of human exposure [45]. Guidelines that can be used to appropriately address the risks involved in using an MBCA are still lacking. More research is needed on the development of test systems and guidelines to adequately measure the risk a microorganism poses to the environment [47]. The methodology developed by Van Lenteren et al. [48] to assess the risks of import and release of exotic enemies used in inundative forms of biological control cannot be extrapolated for MBCAs [47, 49]. Längle [50] further developed the model that integrates information on the potential of an organism to establish and disperse, its host range and possible direct and indirect effects on nontarget organisms, to better fit the needs for MBCA risk assessment. All the above mentioned requirements for MBCAs have significantly contributed to the increase in the costs for obtaining a registration. A decade ago, the costs for registration of an MBCA were estimated at $ in the US and Canada with a total of $1 2 million for its development, registration and commercialization [11, 51]. However, more recent estimates were in the range of $ to 1 million for the EPA registration process [9]. Moreover, the costs for obtaining a registration in a single EU member state for on a specific crop can reach $ [37]. Krause et al. [33] calculated that based on the current EU registration requirements an investment of $7 9 million over the course of 6 10 years is needed for registering and commercializing a new MBCA in the EU. Therefore, before a decision is taken on the commercialization of biocontrol product a thorough market analysis is necessary. The development should be market- and not product-driven to avoid failure and ensure that a return on investment is possible within the first three years after sales have begun [7]. Outlook Although almost all the companies that undertook registrations of the products listed in (Table 2) have currently suspended their research and development efforts for new biocontrol agents, there are still many reasons for being optimistic. There are several initiatives underway that will help to promote the use of biocontrol products in the future. Concerning efficacy, a certification scheme was developed that provides a farmer with data on the efficacy of an MBCA to counteract the impression of snake oil : products that are sold as biopesticides but without proof of efficacy [6]. From the regulators side there have been new initiatives to promote the use of alternative products. The Pesticide Safety Directorate (PSD) in the UK has launched a new biopesticide scheme to facilitate more alternative products entering the market. Key elements are the appointment of a Biopesticide Champion, an expert providing the initial contact to help applicants through the application process. Secondly, specific guidance to applicants is provided via pre-submission meetings. Thirdly, the cost of evaluations, which has been one of the main concerns for applicants, has been reduced [21] (for more details see Currently, the EU funded specific support action, acronym REBECA (Regulation of Biocontrol Agents), is underway with the objective of bringing together stakeholders from industry, science, regulatory authorities, policy and environment to form a network of expertise within the EU (for more details see This expertise should help to improve regulatory procedures for MBCAs [52]. With this in mind, the use of MBCAs can play an important role in future crop protection, as a key element in IPM programmes. References Sebastian Kiewnick 9 1. Copping LG, Menn JJ. Biopesticides: a review of their action, applications and efficacy. Pest Management Science 2000; 56: Warrior P. Opportunities and challenges for microbial pesticides in the global market place. Phytopathology 2006;96 Suppl.: Anderson J. EPA s role in biopesticide development. 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