U.S. regulatory framework for genetic biocontrol of invasive fish

This paper provides an overview of the U.S. regulatory framework governing genetic biocontrol efforts for invasive fish. Genetic biocontrol refers to the intentional release of genetically modified organisms (GMOs) into the environment to control a target population of a non-native species. The terms “genetically modified” and “genetically engineered” are often used interchangeably, despite the scientific distinctions. A GMO is an organism that has had its genetic material altered or modified by humans through any method, including conventional breeding. Genetic engineering, as defined by the Food and Drug Administration (FDA), is the use of recombinant DNA techniques to introduce new characteristics or traits into an organism. GE organisms are therefore a subset of GMOs. As this paper will discuss, existing laws focus on GE organisms raising significant questions as to whether organisms modified without utilizing rDNA techniques fall within the jurisdiction of any federal agency. Under the 1986 Coordinated Framework for Regulation of Biotechnology, three federal agencies have primary responsibility over biotechnology—the Environmental Protection Agency (EPA), the U.S. Department of Agriculture, and the FDA. Because the EPA has exempted biological control agents from regulation as pesticides and no fish species are currently considered plant pests, the FDA is the agency responsible for approving the use of genetically engineered fish for biocontrol. FDA regulates genetically engineered animals through its New Animal Drug Application (NADA) process. The NADA process presents several challenges to effective and transparent regulation of genetic biocontrol, including the FDA’s focus on drug safety, secrecy provisions potentially limiting disclosure of the results of environmental reviews, and the secondary role of the Fish and Wildlife Service, the federal agency with the most experience with invasive species management. In addition, relying on the NADA process creates a significant regulatory gap as NADA approval is only required for GE organisms. The regulatory framework for GMOs created for genetic biocontrol without rDNA technology is unclear and primary responsibility may fall to the states. Given its extensive experience with hatcheries, invasive fish species control, and environmental reviews, the Fish and Wildlife Service (FWS) is the more appropriate agency to review applications for genetic biocontrol. Efforts should be undertaken now, while genetic biocontrol is still in the theoretical stages, to increase the role of the FWS in the permitting process either through formal regulations or more informal mechanisms such as memorandum of understanding.     

Multicellular behaviour and production of a wide variety of toxic substances support usage of Bacillus subtilis as a powerful biocontrol agent

Intensive cultivation of plants in the monoculture field system in order to feed the continuously growing human population creates a need for their protection from the variety of natural competitors such as: bacteria, fungi, insects as well as other plants. The increase in the use of chemical substances in the 20th century has brought many effective solutions for the agriculture. However, it was extremely difficult to obtain a substance, which would be directed solely against a specific plant pathogen and would not be harmful for the environment. In the late 1900's scientists began trying to use natural antagonisms between resident soil organism to protect plants. This phenomenon was named biocontrol. Biological control of plants by microorganisms is a very promising alternative to an extended use of pesticides, which are often expensive and accumulate in plants or soil, having adverse effects on humans. Nonpathogenic soil bacteria living in association with roots of higher plants enhance their adaptive potential and, moreover, they can be beneficial for their growth. Here, we present the current status of the use of Bacillus subtilis in biocontrol. This prevalent inhabitant of soil is widely recognized as a powerful biocontrol agent. Naturally present in the immediate vicinity of plant roots, B. subtilis is able to maintain stable contact with higher plants and promote their growth. In addition, due to its broad host range, its ability to form endospores and produce different biologically active compounds with a broad spectrum of activity, B. subtilis as well as other Bacilli are potentially useful as biocontrol agents.     

Melding ecology,classical weed biocontrol,and plant microbial ecology can inform improved practices in controlling invasive plant species

Early research leading to the successful biological control of invasive species such as Opuntia spp., and Hypericum perforatum set examples and provided data useful for research programs that would follow. However, this early work failed to become established as a source of applicable principles for later workers in weed biocontrol. Recently, retrospective and parallel studies have been suggested as a means to reengage with earlier work to derive useful ideas and data to enhance future programs in weed biocontrol. Parallel studies by workers in plant community ecology on the nature of feedback elicited by plant species in their invaded and native range have shown the importance of soil microbial communities in effecting feedback. Retrospective reexamination of previous studies would likely provide clues to other insect–plant pathogen interactions in addition to those described by the author and others. The effects of invasive species in profoundly altering soil microbial communities point to the need for further studies on key microbial species contributing to or driving the impact of biocontrol. These collective data suggest that the desired goal of selecting for and utilizing stronger biocontrol agents to reduce nontarget effects and to increase the impact of biological control programs would be best served by prerelease studies that assess the propensity of a candidate agent for direct or indirect interaction with other agents. This could be assessed through the use of survival analysis. Overall, parallel empirical and retrospective studies should be a necessary part of how biological control is practiced.     

Melding ecology, classical weed biocontrol, and plant microbial ecology can inform improved practices in controlling invasive plant species

Early research leading to the successful biological control of invasive species such as Opuntia spp., and Hypericum perforatum set examples and provided data useful for research programs that would follow. However, this early work failed to become established as a source of applicable principles for later workers in weed biocontrol. Recently, retrospective and parallel studies have been suggested as a means to reengage with earlier work to derive useful ideas and data to enhance future programs in weed biocontrol. Parallel studies by workers in plant community ecology on the nature of feedback elicited by plant species in their invaded and native range have shown the importance of soil microbial communities in effecting feedback. Retrospective reexamination of previous studies would likely provide clues to other insect–plant pathogen interactions in addition to those described by the author and others. The effects of invasive species in profoundly altering soil microbial communities point to the need for further studies on key microbial species contributing to or driving the impact of biocontrol. These collective data suggest that the desired goal of selecting for and utilizing stronger biocontrol agents to reduce nontarget effects and to increase the impact of biological control programs would be best served by prerelease studies that assess the propensity of a candidate agent for direct or indirect interaction with other agents. This could be assessed through the use of survival analysis. Overall, parallel empirical and retrospective studies should be a necessary part of how biological control is practiced.     

Genotypic comparison ofPantoea agglomeransplant and clinical strains

Background  

Pantoea agglomeransstrains are among the most promising biocontrol agents for a variety of bacterial and fungal plant diseases, particularly fire blight of apple and pear. However, commercial registration ofP. agglomeransbiocontrol products is hampered because this species is currently listed as a biosafety level 2 (BL2) organism due to clinical reports as an opportunistic human pathogen. This study compares plant-origin and clinical strains in a search for discriminating genotypic/phenotypic markers using multi-locus phylogenetic analysis and fluorescent amplified fragment length polymorphisms (fAFLP) fingerprinting.     

Identification and analysis of differentially expressed cDNAs during nonself-competitive interaction between Phlebiopsis gigantea and Heterobasidion parviporum

The molecular factors regulating interspecific interaction between the saprotrophic biocontrol fungus Phlebiopsis gigantea and the conifer pathogen Heterobasidion parviporum were investigated. We constructed cDNA libraries and used expressed sequence tag analysis for the identification and characterization of genes expressed during the self and nonself-hyphal interaction. cDNA clones from either the pathogen or biocontrol agent were arrayed on nylon membrane filters and differentially screened with cDNA probes made from mycelia forming the barrage zone during nonself-interactions, mycelia growing outside the barrage zones or monocultures. BlastX analysis of the differentially expressed clones led to the identification of genes with diverse functions, including those with potential as virulence factors, such as hydrophobins. Because of the high sequence conservation (r2 = 0.81) between P. gigantea and H. parviporum, a selected number of genes from either fungus were used to monitor the expression profile under varying interaction conditions by virtual northern blot. The results are discussed with respect to the potential role of the induced genes during the nonself-competitive interaction for space and nutrients between P. gigantea and H. parviporum.     

Current and Future Use of Nematodes in Biocontrol: Practice and Commercial Aspects with Regard to Regulatory Policy Issues

Constraints about the use of chemical insecticides have limited the availability of control measures against soil-borne insect pests. Entomopathogenic nematodes of the genera Steinernema and Heterorhabditis provide an environmentally safe and economically reasonable alternative. Their life cycle and current production, storage and formulation technology are described. An overview of their safety, use in integrated pest management and current market potential (US$10 million in 1994) is also given. The costs of research and development efforts and the scale-up of production technologies are discussed in relation to the current and future market potential. Large-scale, outdoor application will require additional scientific and technical progress in the areas of production, storage, formulation and application. Besides public funding, the current niche markets will need to provide the financial basis for further development, provided that regulatory conditions will not limit the exploitation of the nematodes' market potential. It is recommended that nematodes should be exempted from registration. Rules for risk assessment in the use of exotic nematodes should be internationally harmonized and related specifically to the biology and ecology of these nematodes. The volume of current markets would not justify the costs of registration procedures currently required for chemical or microbial insecticides or genetically engineered organisms. Regulatory policies should aim at supporting the further introduction of entomopathogenic nematodes as biocontrol agents.     

盾壳霉在油菜菌核病菌生物防治中的应用

油菜核病菌(Sclerotiniasclerotiorum)是一种世界性病原菌,其分布广、危害大、难根治。盾壳霉(Coniothyriumminitans)是该病原菌的破坏性寄生真菌,可以有效、专一地降低病原菌菌核的形成与萌发,在该病原菌的生物防治方面具有较大的应用潜力。从油菜核盘菌的致病过程与特点、盾壳霉的生长特性、盾壳霉和油菜核盘菌间相互作用的规律及途径等几个方面阐述了盾壳霉对油菜核盘菌的生防特性,讨论了盾壳霉在生产实践中的应用潜力及存在问题,并提出了一些解决问题的可能途径及需要进一步研究的内容与方向 。     

Biological control of fungal plant diseases

Research has demonstrated the agricultural potential of biological control. For airborne pathogens as well as for soilborne pathogens similar strategies based on different targets in the life cycle of the pathogen can be distinguished, viz. (1) microbial protection of the host against infection, (2) microbial reduction of pathogen sporulation and (3) microbial interference with pathogen survival. Some successes and failures with respect to these targets will be discussed and include (1) biocontrol of seedling diseases, root pathogens, and post-harvest diseases (2) biocontrol of powdery mildew and Botrytis cinerea (3) biocontrol of sclerotial pathogens. Despite of a lot of research on biological control of plant diseases, the number of products available is limited and their market size is marginal. The market for biological control products is not only determined by agricultural aspects such as the number of diseases controlled by one biocontrol product in different crops but also by economic aspects as cost-effective mass production, easy registration and the availability of competing means of control including fungicides. The future development of low-chemical input sustainable agriculture and organic farming will determine the eventual role of biological control in agriculture.     

Using evolutionary tools to facilitate the prediction and prevention of host‐based differentiation in biological control: a review and perspective

The unprecedented success of biological control (biocontrol) agents led some of the proponents of this technology to promote its use as a panacea for all pest problems. Following an accumulation of non‐target host interactions, because of generalist or new association introductions, techniques to help ensure classical biocontrol agent's success and reduce non‐target interactions were implemented. Even with these new measures in place, public and scientific mistrust and lack of consistency has resulted in increased regulation of biocontrol introductions. This has likely decreased the probability of effective, sustainable control measures being expeditiously implemented. With the current apprehension concerning the safety of biocontrol, we should incorporate the processes (adaptation, selection, etc.) and theoretical concepts of evolutionary biology to predict and enhance the effectiveness of biocontrol. The microevolutionary perspective that involves mutation, drift, selection and gene flow may be a crucial consideration in the realm of biocontrol. Here, we discuss how and why spatial and evolutionary models should be implemented into future risk assessment analyses of potential biocontrol agents. We suggest that it is necessary to re‐assess the approach that has developed over the past approximately 100 years of sustained releases and illuminate them in the context of an evolutionary timescale.     

Use of a breeding approach for improving biocontrol efficacy of Phlebiopsis gigantea strains against Heterobasidion infection of Norway spruce stumps

Sixty-four wild heterokaryotic isolates of Phlebiopsis gigantea were analysed for asexual spore production, growth rate and competitive ability against Heterobasidion in vitro , as well as growth rate in Norway spruce wood. These P. gigantea traits were considered important for controlling infection of Norway spruce stumps by spores of Heterobasidion spp. Ten most promising P. gigantea isolates were crossed with each other and 172 F₁ progeny heterokaryons were analysed for the above-mentioned traits. Thirteen most promising progeny heterokaryons were selected and their biocontrol ability against infection by Heterobasidion was compared with the parental isolates in stem pieces of Norway spruce. The results indicated that the progeny strains had generally better traits and control efficacy than the parental strains. The genetic effects accounted for a part of the variations between progeny and parental strains. This further suggests that there is a potential to improve the biocontrol properties of P. gigantea through breeding.     

Occurrence and impact of the root-rot biocontrol agent Phlebiopsis gigantea on soil fungal communities in Picea abies forests of northern Europe

The aim of this study was to assess belowground occurrence, persistence and possible impact of the biocontrol agent Phlebiopsis gigantea (Fr.) Jülich on soil fungi. Sampling of soil and roots of Picea abies (L.) H. Karst. was carried out at 12 P. gigantea-treated and five nontreated control sites representing 1- to 60-month-old clear-cuts and thinned forest sites in Finland and Latvia. The 454-sequencing of ITS rRNA from fine roots, humus and mineral soil resulted in 8626 high-quality fungal sequences. Phlebiopsis gigantea represented 1.3% of all fungal sequences and was found in 14 treated and nontreated sites and in all three substrates. In different substrates, the relative abundance of P. gigantea at stump treatment sites either did not differ significantly or was significantly lower than in nontreated controls. No significant correlation was found between the time elapsed since the tree harvesting and/or application of the biocontrol and abundance of P. gigantea in different substrates. In conclusion, the results demonstrate that P. gigantea occasionally occurs belowground in forest ecosystems but that stump treatment with the biocontrol agent has little or no impact on occurrence and persistence of P. gigantea belowground, and consequently no significant impact on soil fungi.     

Soilborne Plant Diseases Caused by Pythium spp.: Ecology,Epidemiology, and Prospects for Biological Control

Soilborne root diseases caused by plant pathogenic Pythium species cause serious losses in a number of agricultural production systems, which has led to a considerable effort devoted to the development of biological agents for disease control. In this article we review information on the ecology and biological control of these pathogens with the premise that a clear understanding of the ecology of the pathogen will assist in the development of efficacious biocontrol agents. The lifecycles of the pathogens and etiology of host infection also are reviewed, as are epidemiological concepts of inoculum-disease relationships and the influence of environmental factors on pathogen aggressiveness and host susceptibility. A number of fungal and bacterial biocontrol agents are discussed and parallels between their ecology and that of the target pathogens highlighted. The mechanisms by which these microbial agents suppress diseases caused by Pythium spp., such as interference with pathogen survival, disruption of the process of plant infection, and induced host resistance, are evaluated. The possibilities for enhancement of efficacy of specific biological control agents by genetic manipulation or deployment tactics are discussed, as are conceptual suggestions for consideration when developing screening programs for antagonists.     

Ecological,practical, and political inputs into selection of weed targets: What makes a good biological control target?

The topic of ecological, practical, and political considerations in the selection of weed targets for biological control has been widely discussed during the past two decades, mostly from the perspective of insect herbivores. For conceptual and practical purposes, plant pathogens have been treated in these discussions as if they are a subset of inoculative biocontrol agents, with little said about the inherent differences between pathogens and insects as biocontrol agents or the selection of weed targets for control by the inundative, bioherbicide strategy. Herein, I attempt to address the question of what makes a good biological control target for plant pathogens used as inoculative as well as inundative agents, basing my analysis on examples from the past three decades. Despite the small number of examples available for this analysis, the following generalizations can be made: (1) Weeds with robust capacity for vegetative regeneration are more difficult to control with pathogens than those that lack this trait. (2) A plant’s growth habit is not a reliable guide for target selection; weeds that have been successfully controlled include annual and biennial herbs, perennial shrubs, perennial vines, and trees, while numerous failures have been reported irrespective of the target’s growth habit or reproductive mode. (3) It is more challenging to control species with genetic heterogeneity and capacity for introgression than genetically homogeneous and reproductively conserved species. (4) Matching the target host’s susceptibility with the candidate pathogen’s virulence is of utmost importance for biocontrol success since host–pathogen interactions at the species and subspecies levels are often governed by single-gene differences (e.g., varietal specificity). (5) Practical and political considerations are central to the selection of targets for control with pathogens. (6) Demand from influential stakeholders for control and/or for a nonchemical or economically sustainable control typically drives the initiative as well as the continuance of biocontrol projects to their completion. (7) In the case of inundative, bioherbicide agents, the continuity and ultimate implementation of a project will be dictated by the prospects of economic returns from developing and using a pathogen. (8) The stakeholders’ perceptions of the effectiveness of a biocontrol program can be unpredictable, leading to conflicting views of “success.” In the final analysis, a good weed target for control by a pathogen is one that has strong stakeholder backing and the list of available pathogens for the target suggests a possibility of acceptable control at a cost that is competitive with those of other control options. While this conclusion is also applicable to target selection for insect biocontrol agents, it is more relevant for pathogens because of limited funding and personnel available for development of pathogens and the added cost and technological complexity of implementing bioherbicides compared to classical biocontrols.     

Inhibition of fungal spore germination by gramicidin S and its potential use as a biocontrol against fungal plant pathogens

Gramicidin S, as well as being sporicidal to Bacillus spores, also inhibits germination and emergence of fungal-like spores of Dictyostelium discoideum . The fungal plant pathogen Fusarium nivale is also inhibited and gramicidin S, therefore, is a sporicidal and antifungal antibiotic. Considering these findings the potential use of this antibiotic and its producer organism Bacillus brevis as a biocontrol is discussed.     

Experts know more than just facts: eliciting functional understanding to help prioritise weed biological control targets

Prioritising investments in classical weed biological control (biocontrol) is a common decision-making challenge: biocontrol programmes can yield substantial benefits but are typically long-term and costly, and the outcome uncertain. Experts are often relied upon to help, but their role is generally restricted to providing facts and judgements to populate an existing prioritisation model, which in turn receives little scrutiny. We developed and applied a new prioritisation framework to guide biocontrol investment decisions by livestock industries that required eliciting experts’ functional understanding (including their in-depth knowledge of the theoretical and practical drivers of weed biocontrol programmes). This consultative and transparent framework drew on expertise from most biocontrol practitioners in Australia through a structured workshop, and the literature. Each of the 75 weed taxa considered was placed in a matrix according to their impact (current or potential) and the prospects of biocontrol achieving pre-defined management goals. There was considerable knowledge uncertainty regarding potential impacts, which is of concern when making pre-emptive investments. Feasibility (likelihood of finding host-specific agents) and likelihood of success (management goals being met, assuming that host-specific agents are available) of biocontrol were both assessed as low for 51 % of taxa. Predicted barriers to successful biocontrol were diverse and idiosyncratic, suggesting that application of more quantitative prioritisation approaches would be challenging. A short-list of 13 weed taxa was identified for further consideration as biocontrol targets, based on the trade-off between potential impact and prospects for biocontrol. Research priorities emerged from the prioritisation process that would maximise investment outcomes for each taxon. Only two short-listed taxa are new targets, reflecting the maturity of the biocontrol discipline targeting weeds of livestock industries in Australia. Accessing the in-depth functional understanding of experts resulted in explicit characterisation of the barriers to successful biocontrol and if/how they might be overcome, improved characterisation of uncertainty, and provided directed guidance for investment. Such an approach would be readily applicable to analogous decision-making challenges in other sectors and countries.     

Potato Cultivar, Pathogen Isolate and Antagonist Cultivation Medium Influence the Efficacy and Ranking of Bacterial Antagonists of Fusarium Dry Rot

The process of selecting biological control agents for further development frequently does not involve conducting bioassays of strain effectiveness on a range of pathogen isolates or host cultivars. Additionally, though previous studies have demonstrated that the medium used to produce biomass of an antagonist can alter its efficacy, this factor is also rarely considered when selecting for the most effective antagonist. Host cultivar, pathogen isolate, and the cultivation medium used to produce the antagonists' biomass were examined as factors of potential importance for assessing the relative effectiveness of bacterial biocontrol strains accurately. Five bacterial antagonists that control Fusarium dry rot on stored potato tubers were assayed for effectiveness against 10 isolates of Gibberella pulicaris . All antagonists reduced disease severity (35-81%) regardless of the specific assays conducted. However, when the antagonists' biomass were produced on two media that differed both in nutrient composition and phase, the efficacy ranking of antagonist Enterobacter sp. S11:P:08 varied from first to fourth most effective. For the antagonists studied, the phase of a nutritionally identical medium had little impact on the efficacy ranking of the five antagonists. Four of the five antagonists had efficacy rankings that ranged from first to last depending on the isolate of the pathogen used to conduct the bioassay. The cultivar of the host also caused variations in the efficacy ranking of the antagonists. These results indicate that bioassays should be conducted using a range of liquid culture production media, pathogen isolates, and host cultivars in order to choose an antagonist that has the highest likelihood for commercial development as an effective biological control product.     

Use of Risk Analysis for Screening Weed Biocontrol Agents: Altica carduorum Guer. (Coleoptera: Chrysomelidae) from China as a Biocontrol Agent of Cirsium arvense (L.) Scop. in North America

Candidate weed biocontrol agents must be screened to exclude those that could threaten desirable plants . Traditionally , this has been done by rejecting species that develop on economically important plants in laboratory no - choice tests . However , because congeneric plants often support development in these tests , even when they are not utilized in nature , the tests do not meet legislated requirements for rare plant species or the increasing public concern for native plants . Plant suitability for larval development is a poor predictor of host range because insects use a sequence of steps in which the early steps , such as host finding and acceptance for oviposition , tend to be stronger than the later ones , such as suitability for development . This study is a trial of a new approach to screening insects as weed biocontrol agents that uses risk analysis to quantify the suitability of a plant as a host on the basis of inset performance at various stages in its life cycle . The insects used for the study was a NW Chinese biotype of the leaf beetle , Altica carduorum, which in terms of climate adaptation and damage it inflicts on the weedy thistle Cirsium arvense, is a promising biocontrol agent for Canada . However , its ability to develop on all North American Cirsium spp . in laboratory no - choice tests currently excludes its release . We show by risk and factor analyses , with five sequential host - selection parameters , that the suitability of these thistles to A. carduorum is so low that the beetle would not jeopardize the continued existence of rare native thistles , so its release should not be a problem .     

GFP technology for the study of biocontrol agents in tritrophic interactions: a case study with Pseudozyma flocculosa

GFP technology was applied to the biocontrol agent (BCA) Pseudozyma flocculosa to study its development and interactions at the tritrophic level plant-powdery mildew-BCA. Transformation experiments with GFP led to the production of a strongly fluorescent strain, Act-4, that displayed biocontrol traits typical of P. flocculosa WT. Following inundative applications, growth of P. flocculosa Act-4 was closely and almost exclusively associated with the colonies of the pathogen regardless of the powdery mildew species or the host plant tested. Development of P. flocculosa Act-4 on control leaves alone was extremely limited 24 h after its application and was typical of the epiphytic growth characterizing this type of yeast-like fungus. Based on the strong correlation between the colonization pattern of the different powdery mildew species tested and the presence of P. flocculosa Act-4, as determined by its fluorescence, it seems that growth of the BCA is dependant on the presence of powdery mildews. These results demonstrate that the GFP technology can be used to study plant-pathogen-BCA interactions and fulfill a wide array of purposes ranging from fundamental observations of the biocontrol behavior of a BCA to very applied ones serving some of the requirements for the registration of BCA's such as defining their environmental fate.     

The effects of gibberellin A3 on white clover during floral development

Hokkanen & Pimentel (1984) proposed a novel approach for the selection of biocontrol agents. They advocated the selection of agents from a relative of the weed plant rather than from the target weed species itself. The new relationship that such agents would have with the weed would be characterised by a relative lack of homeostasis compared with the old herbivore-plant associations traditionally used in weed biocontrol, and would consequently be more effective. There are few examples to support these contentions because of the traditional use of old herbivore-plant associations in weed biocontrol. In the present study, herbivore-plant associations in agricultural situations, which are analogous to agent-weed associations, are examined to assess the potential of new associations for weed biocontrol. The herbivores on 14 introduced crop plants which have salient similarities to the major weeds in the south-western Cape were surveyed: (a) 68% of the 188 insect and mite herbivores are indigenous species in new associations with these host plants, and (b) of the five most damaging pests on each of 13 of the crop plants, 53% are in new associations with the plants. Of the 40 most important agricultural pests in South Africa, 58% are in new associations, confirming these results. About 50% of the insect and mite herbivores in new associations with their host plants are oligophagous, indicating that new associations are not necessarily characterised by polyphagy and hence unsafe for use in weed biocontrol. We conclude that new associations between herbivore species and host plants have strong potential in weed biocontrol because (a) their frequency in agriculture indicates that they can easily be established in weed biocontrol situations, (b) they are as damaging as old associations and (c) they are not necessarily unsafe as regards host specificity. We therefore advocate the use of both the classical approach and that of Hokkanen & Pimentel (1984). Our support for Hokkanen & Pimentel (1984) is, however, based on evidence and rationale clearly different to theirs, and we provide novel guidelines, which can be routinely and practically applied in the selection of agents for weed biocontrol.