Scientific advances in the analysis of direct risks of weed biological control agents to nontarget plants

Research on host specificity testing protocols over the last 10 years has been considerable. Traditional experimental designs have been refined and interpretation of the results is benefiting from an improved understanding of agent behavior. The strengths, weaknesses, and best practice for the different test types are now quite clearly understood. Understanding the concept of fundamental host range (the genetically determined limits to preference and performance) and using this to maximize reliability in predicting field host specificity following release (behavioral expression of the fundamental host range under particular conditions) are still inconsistently understood or adopted despite having been identified as the critical steps in analyzing the threats posed by biological control agents to the agriculture and biodiversity of novel environments. This needs to be consistently understood and applied so the process of testing can follow a recognized process of risk analysis from hazard identification (identifying life stages of the agent that pose a threat and defining their fundamental host range) to uncertainty analysis based on the magnitude (predicted field host specificity following release) and likelihood of threats (predicted actual damage and impact) to nontargets. Modern molecular techniques are answering questions associated with subspecific variation in biological control agents with respect to host use and the chance of host shifts of agents following release. Guidelines for assessment of nontarget impacts need to recognize and adopt such recent developments and emphasize a general increased understanding of the evolution of host choice and the phylogenetic constraints to shifts in host use. This review covers all these recent advances for the first time in one document, highlighting how inconsistent interpretation by biological control practitioners can be avoided.     

Native-range research assists risk analysis for non-targets in weed biological control: the cautionary tale of the broom seed beetle

Abstract  This case study considers the broom seed beetle, Bruchidius villosus , a narrowly oligophagous species within the Fabaceae, subtribe Genistinae for which in-depth native-range studies have been vital to help understand the likely field host specificity following release. Bruchidius villosus has been used in three countries as a classical biological control agent against Scotch broom, Cytisus scoparius . Original host-specificity testing of a UK population, where this species had only been observed developing on C. scoparius , suggested this population was specific to the target. The beetle was released in New Zealand. Following release, however, the agent exhibited a broader host range than in the tests, but not a broader host range than that of the species as a whole. Subsequent studies in the native range using surveys and field testing have helped to show why B. villosus populations exhibit higher specificity in the native range than would be expected from the species' host range. This case is used to illustrate the contribution native-range studies can make to science-based risk analysis of biological control agents against weeds. By doing so, they also highlight the associated risks of ignoring native-range studies and adopting of a 'grab-and-run' approach to obtaining classical biological control agents.     

Statistical tools to interpret risks that arise from rare events in host specificity testing

Host specificity testing to predict host range is one of the key steps to predicting the risk a biological control agent will present to non-target organisms in the new environment. When host specificity testing data contain discrepancies, or unacceptable levels of uncertainty, it can be difficult for decision-makers to adequately address this uncertainty. To better understand the uncertainty in host specificity testing, we used a range of statistical tools to examine a data set associated with the leaf weevil Cleopus japonicus (Curculionidae), a biological control agent for the weed Buddleja davidii (Buddlejaceae) in New Zealand. Significant uncertainty arose during the early stages of host specificity testing when one C. japonicus larva reared to pupation on a culturally important native plant. Further trials were conducted to evaluate the suitability of C. japonicus as a biological control agent, and despite the uncertainty, C. japonicus was released in New Zealand in 2006, and has since established populations at each release site. However, the possibility of larvae completing their life cycle on the native plant initiated this evaluation of the statistics associated with testing biological control agents. We present results from analyses of the C. japonicus survival data using confidence intervals, equivalence testing, power analyses and survival curves to highlight the appropriateness of each of these tools for interpreting host specificity tests in biological control.     

Is host‐specificity of weed biological control agents likely to evolve rapidly following establishment?

Over 352 herbivore species have now been intentionally introduced into new regions as weed biological control agents. Recent evidence shows that rapid and significant evolution in host-specificity can occur. The risk of non-target use by biological control agents increasing to unacceptable levels through rapid evolution therefore needs to be considered. In addition, weed biological control offers many as yet largely unexploited opportunities for improving our basic understanding of host-specificity and its evolution. We therefore evaluate the evidence that rapid evolution (1) alters the use of existing hosts, and (2) alters the fundamental host-range. Most cited examples of so-called host shifts from weed biological control were not the result of genetic change. There was only limited evidence of genetically altered performance on a non-target host and no evidence of altered fundamental host-range. We conclude, from both theory and the available data, that only altered use of existing hosts (through quantitative genetic changes) needs be considered when evaluating the risk of rapid evolution. Host-specificity testing methodologies can be improved and adapted to better assess the risk of occurrence of post-release evolution.     

Predictability of pathogen host range in classical biological control of weeds: an update

Before an exotic pathogen can be released as a classical biological control agent the likely positive and negative outcomes of that introduction must be predicted. Host range testing is used to assess potential damage to non-target plants. To-date 28 species of fungi have been released as classical biological control agents against weeds world-wide. These pathogens have been reported infecting only six non-target plant species outdoors and all of these incidents were predicted. Many more non-target plant species developed disease symptoms in glasshouse tests than in the field. Consequently, data from other sources are needed to ensure potential agents are not prematurely rejected. Predictions of pathogen host range to date have been sufficiently accurate to prevent unpleasant surprises. Exotic pathogens are a safe and useful tool for weed control, especially in natural areas rich in valued non-target species.     

外来入侵杂草传统生物防治的生态风险及其防范对策

长期大量实践说明,引进天敌防治外来入侵杂草的传统生物防治方法是治理外来入侵杂草的一条切实可行的有效途径,但对其潜在的生态风险——对本土生物的直接或间接不良影响不容忽视。利用传统评价方法预测候选天敌的生态风险存在缺陷,主要表现在：（1）寄主专一性测定过分依赖室内进行的生理寄主范围测定结果,对生态寄主范围（实际寄主范围）问题重视不够,后者指在新环境中的一系列物理和生物条件下的寄主利用预测;（2）在生理寄主范围测定中,过分依赖完成生长发育的可能性,对行为、遗传性状以及系统发育关系重视不够;（3）在风险评估中,过多强调对经济作物的风险,而对自然生态系统的风险重视不够。对此,建议：（1）鼓励对已释放的天敌进行回顾性跟踪研究,从而为杂草生物防治实践提供生态学理论支撑;（2）在运用生物防治手段对付外来入侵杂草实践中,建议采用“有害推论”的预防性原则,以避免在面临入侵生物重大威胁时草率做出释放天敌的决策;（3）在评估候选天敌风险中重视生态效应的风险评估。     

A proposal to use plant demographic data to assess potential weed biological control agents impacts on non-target plant populations

Weed biocontrol programs aim to reduce the spread and population growth rate of the target plant while stabilizing or increasing populations of those native species considered under threat by invasive plants. This goal is not unique to weed biocontrol but applies to all other invasive plant management techniques, though such information is rarely collected. Without this information, success of management interventions can be ambiguous, and regulatory agencies, the public, policy makers, funders and land managers cannot be held accountable for chosen treatments. A fundamental reform, including use of demographic studies and long-term assessments, are essential to guide weed biocontrol programs. We propose to add use of plant demography (an assessment of how environmental factors and ecological interactions, for example competition, disease or herbivory, may affect plant populations by altering survival, growth, development and reproductive rates of plant individuals) during host specificity risk assessments of potential biological control agents. Demographic models can refine assessments of potential impacts for those plant species that experience some feeding or larval development during host specificity testing. Our proposed approach to focus on impact on plant demography instead of attack on plant individuals is useful in appropriately gauging threats potential weed biocontrol agents may pose to non-target species after field release.     

A systematic approach to biological control agent exploration and prioritisation for prickly acacia (Acacia nilotica ssp. indica)

Abstract  Agent selection for prickly acacia has been largely dictated by logistics and host specificity. Given that detailed ecological information is available on this species in Australia, we propose that it is possible to select agents based on agent efficacy and desired impact on prickly acacia demography. We propose to use the 'plant genotype' and 'climatic' similarities as filters to identify areas for future agent exploration; and plant response to herbivory and field host range as 'predictive' filters for agent prioritisation. Adopting such a systematic method that incorporates knowledge from plant population ecology and plant–herbivore interactions makes agent selection decisions explicit and allow more rigorous evaluations of agent performance and better understanding of success and failure of agents in weed biological control.     

Cryptic species of a water hyacinth biological control agent revealed in South Africa: host specificity,impact, and thermal tolerance

The discovery that cryptic species are more abundant than previously thought has implications for weed biological control, as there is a risk that cryptic species may be inadvertently released with consequences for the safety of the practice. A cryptic species of a biological control agent released for the control of the invasive alien macrophyte, water hyacinth, Eichhornia crassipes (C. Mart.) Solms. (Pontederiaceae), was recently discovered in South Africa. The two species were considered a single species prior to genetic analysis and interbreeding experiments. The original biological control agent retains the name Eccritotarsus catarinensis (Carvalho) (Heteroptera: Miridae) whereas the new species has been described as Eccritotarsus eichhorniae Henry. In this study, we compared the host specificity, efficacy, and thermal physiologies of the two species. The host specificity of the two species within the Pontederiaceae was very similar and both are safe for release in South Africa. Comparison of the per capita impact of the two species indicated that E. eichhorniae was the more damaging species but this is likely to be influenced by temperature, with E. catarinensis being more effective under lower temperatures and E. eichhorniae being more effective under higher temperatures. Releasing the correct species for the thermal environment of each release site will improve the level of control of water hyacinth in South Africa. This example highlights the need to keep populations of biological control agents from different native range collection localities separate, and to screen for host specificity and efficacy.     

Environmental risk assessment of exotic natural enemies used in inundative biological control

In the past 100 years many exotic naturalenemies have been imported, mass reared andreleased as biological control agents. Negativeenvironmental effects of these releases haverarely been reported. The current popularity ofinundative biological control may, however,result in problems, as an increasing number ofactivities will be executed by persons nottrained in identification, evaluation andrelease of biological control agents.Therefore, a methodology for risk assessmenthas been developed within the EU-financedproject `Evaluating Environmental Risks ofBiological Control Introductions into Europe[ERBIC]' as a basis for regulation of importand release of exotic natural enemies used ininundative forms of biological control (i.e.not in `classical biological control' thoughsome of the same principles and approachesapply). This paper proposes a general frameworkof a risk assessment methodology for biologicalcontrol agents, integrating information on thepotential of an agent to establish, itsabilities to disperse, its host range, and itsdirect and indirect effects on non-targets. Ofthese parameters, estimating indirect effectson non-targets will be most difficult, asmyriads of indirect effects may occur whengeneralist natural enemies are introduced. Theparameter `host range' forms a central elementin the whole risk evaluation process, becauselack of host specificity might lead tounacceptable risk if the agent establishes anddisperses widely, whereas, in contrast, amonophagous biological control agent is notexpected to create serious risk even when itestablishes and disperses well. Drawing onpublished information and expert opinion, theproposed risk assessment methodology is appliedto a number of biological control agentscurrently in use. These illustrative casehistories indicate that the risk assessmentmethodology can discriminate between agents,with some species attaining low `risk indices'and others scoring moderate or high. Riskindices should, however, not be seen asabsolute values, but as indicators to which ajudgement can be connected by biologicalcontrol experts for granting permission torelease or not.     

Ecological role of control agent,and not just host‐specificity,determine risks of biological control

Biological control agents used to manage alien vegetation are generally viewed as providing an ecosystem service, owing to reduced ecological and economic costs of invasion following their release. In particular, gall‐formers are popular as biological control agents because they are host‐specific and therefore considered low risk. However, galls can also be considered to be ecological engineers, because they provide nutritional resources for native invertebrates. We tested whether native invertebrates had formed associations with the gall‐forming fungus Uromycladium tepperianum, introduced into South Africa to control the Australian invasive alien tree Acacia saligna, by collecting U. tepperianum galls and monitoring emergence. We found that a number of invertebrates had formed associations with the biological control agent, among which was the important citrus pest, Thaumatotibia leucotreta (false codling moth). We used pheromone‐baited traps to ascertain if this supplementary source of T. leucotreta increased their abundance in orchards close to patches of gall host, but did not find this to be the case. We did find, however, that control measures used by farmers explained T. leucotreta abundances in traps, which may have obscured detection of any effects of a nearby host for the pest. Nevertheless, this study illustrates the first case of a host‐specific classical biological control agent providing resources for an economically significant crop pest. We conclude that although biological control agents are strictly vetted to ensure host‐specificity, introduced biological control agents that become abundant and can act as ecological engineers pose risks when native biota form associations with them, resulting in a number of possible cascading ecosystem effects. In addition, there could be economic consequences when these associated species include agricultural pests. We conclude that not just host specificity, but potential ecological effects of biological control agents, should be considered in their selection.     

Host-plant specificity and specialization in eriophyoid mites and their importance for the use of eriophyoid mites as biocontrol agents of weeds

Eriophyoid mites, which are among the smallest plant feeders, are characterized by the intimate relationships they have with their hosts and the restricted range of plants upon which they can reproduce. The knowledge of their true host ranges and mechanisms causing host specificity is fundamental to understanding mite-host interactions, potential mite-host coevolution, and diversity of this group, as well as to apply effective control strategies or to use them as effective biological control agents. The aim of this paper is to review current knowledge on host specificity and specialization in eriophyoid mites, and to point out knowledge gaps and doubts. Using available data on described species and recorded hosts we showed that: (1) 80% of eriophyoids have been reported on only one host species, 95% on one host genus, and 99% on one host family; (2) Diptilomiopidae has the highest proportion of monophagous species and Phytoptidae has the fewest; (3) non-monophagous eriophyoids show the tendency to infest closely related hosts; 4) vagrant eriophyoids have a higher proportion of monophagous species than refuge-seeking and refuge-inducing species; (5) the proportions of monophagous species infesting annual and perennial hosts are similar; however, many species infesting annual hosts have wider host ranges than those infesting perennial hosts; (6) the proportions of species that are monophagous infesting evergreen and deciduous plants are similar; (7) non-monophagous eriophyoid species have wider geographic distribution than monophagous species. Field and laboratory host-specificity tests for several eriophyoid species and their importance for biological control of weeds are described. Testing the actual host range of a given eriophyoid species, searching for ecological data, genetic differentiation analysis, and recognizing factors and mechanisms that contribute to host specificity of eriophyoid mites are suggested as future directions for research.     

Prioritisation of potential agents for the biological control of the invasive alien weed,Pereskia aculeata (Cactaceae), in South Africa

Pereskia aculeata Miller (Cactaceae) is an invasive alien species in South Africa that is native in Central and South America. In South Africa, P. aculeata outcompetes native plant species leading to a reduction in biodiversity at infested sites. Herbicidal and mechanical control of the plant is ineffective and unsustainable, so biological control is considered the only potential solution. Climatic matching and genotype matching indicated that the most appropriate regions in which to collect biological control agents were Santa Catarina and Rio de Janeiro provinces in Southern Brazil. Surveys throughout the native distribution resulted in 15 natural enemy species that were associated with the plant. Field host range data, as well as previous host plant records, were used to prioritise which of the species were most likely to be suitably host specific for release in South Africa. The mode of damage was used to determine which species were most likely to be damaging and effective if released. The most promising species prioritised for further study, including host specificity and impact studies, were the stem-wilter Catorhintha schaffneri Brailovsky & Garcia (Coreidae); the stem boring species Acanthodoxus machacalis Martins & Monné (Cerambycidae), Cryptorhynchus sp. (Curculionidae) and Maracayia chlorisalis (Walker) (Crambidae) and the fruit galler Asphondylia sp. (Cecidomyiidae). By prioritising the potential biological control agents that are most likely to be host-specific and damaging, the risk of conducting host specificity testing on unsuitable or ineffective biological control agents is reduced.     

Gallerucida bifasciata (Coleoptera: Chrysomelidae), a potential biological control agent for Japanese knotweed (Fallopia japonica)

Host specificity of foreign natural enemies are becoming more and more critical in classical biological control programs, as concerns about potential risk from introduced biocontrol agents have been increasing recently. Understanding the insect's fundamental and ecological host ranges is the first step in determining the potential for introduction of an insect to control invasive plants. Japanese knotweed, Fallopia japonica (Houttuyn) Ronse Decraene (Polygonaceae) is an invasive weed in the United States and Europe. A leaf beetle, Gallerucida bifasciata (Coleoptera: Chrysomelidae) is an important natural enemy attacking this plant in Asia. However, its host range records were ambiguous. This study examined the beetle's host specificity through a set of choice and no-choice tests in the laboratory and field in its native China. Gallerucida bifasciata larvae were able to complete development on seven of 87 plant species in larval development tests, while adults fed and oviposited on 10 plants in no-choice tests. Multiple choice tests showed adults strongly preferred Fallopia japonica, Persicaria perfoliata (L.) H. Gross and Polygonum multiflorum Thunb over all other plants. Open field tests and field surveys further revealed that these three species were in its ecological host range. The results of this study suggest that G. bifasciata is a potential promising agent for control of Japanese knotweed in the United States and Europe, although additional host specificity tests and risk assessment should be completed.     

Host specificity of entomophilic nematodes—A review

Entomophilic nematodes can be biologically manipulated either as biological control agents or as biological insecticide agents. The characteristics, mode of application, and degree of specificity for each of these is considered. The importance of host specificity information for nematode parasites potentially useful in controlling insect pests is emphasized. A general discussion of host specificity and the factors influencing host specificity of entomophilic nematodes is presented.Characteristics of nematodes of the genus Heterotylenchus are discussed while particular emphasis is placed on H. autumnalis, a potentially useful biological control agent of the face fly, Musca autumnalis. Greater detail of the bionomics of this nematode, its attributes and disadvantages as a successful control agent are considered. The factors resulting in host specificity of this parasite, especially host resistance, in Musca domestica, Orthellia caesarion, and Ravinia l'herminieri, are encapsulation and melanization.It is emphasized that biological control workers are working with a dynamic, evolving host-parasite system and that really they are faced with the same problems (resistance of the host and target specificity of the applied material) originally and still confronting the toxicologist and economic entomologist.     

The role of pre-release efficacy assessment in selecting classical biological control agents for weeds—applying the Anna Karenina principle

The goals in selecting classical biological control agents for weeds are to identify agents that will be both safe for release and effective in controlling their target plants. The release of ineffective agents should be avoided, as these add to the costs and risks of biological control without contributing to its benefits. While the principles of host-specificity testing and risk assessment for weed biological control agents have been extensively debated and refined, there has been less attention given to assessing the probable efficacy of agents prior to release. This reluctance to undertake pre-release efficacy assessment (PREA) is probably based on concerns that it will both add to the cost of screening biological control agents and introduce a risk of wrongly rejecting effective agents. We used a project simulation model to investigate the implications of using PREA as an additional filter in the agent selection process. The results suggest that, if it can be done at a lower cost than host-specificity testing, the use of PREA as the first filter can make agent selection more cost-effective than screening based on host-specificity alone. We discuss examples of PREA and potential approaches. The impact of biocontrol agents is a function of their range, abundance, and per-capita damage. While it will always be difficult to predict the post-release abundance of biological control agents from pre-release studies, some estimates of potential range can be obtained from studies of climatic adaptation. For agents that affect the vegetative growth or survival of their target weeds, experimental measurement of per-capita damage is feasible and can contribute to a reduction in the numbers of ineffective agents released. The Anna Karenina principle states that success in complex undertakings does not depend on a single factor but requires avoiding many separate causes of failure. We suggest that, in biological control of weeds, the use of agents that are not sufficiently damaging is one such cause that can be partially avoided by the use of pre-release efficacy assessment.     

Can an a priori strategy be developed for biological control? The case of Onopordum spp. thistles in Australia

Abstract  Between 1992 and 2000, seven insect agents were released in Australia for the biological control of Onopordum spp. thistles. This paper describes the protocol used for the selection of these agents, starting with the development of a preliminary strategy, based on the ecology and population dynamics of the target weed. The strategy informed the surveys for natural enemies in the native range of Onopordum , targeting insects that attacked key transitional stages of the weed's life cycle. Ongoing studies of Onopordum populations in both Australia and Europe, plus experimental studies on the ecology, potential impact and preliminary host specificity of the agents, led to the refinement of the strategy and the selection and prioritisation of the agents. It is argued that development of an explicit strategy prior to release should be encouraged, as it forces researchers to revisit the rationale for and aims of particular biological control projects, ensuring that the process of agent selection remains focused. It also provides a tool to improve the process of agent selection, as subsequent results can be measured against the strategy and agent success or failure evaluated against the a priori expectations.     

Transmission as a predictor of ecological host specificity with a focus on vertical transmission of microsporidia

Consideration of vertical transmission is particularly important for understanding the life cycles of entomopathogens that are naturally occurring in invertebrate populations, are a problem in beneficial insect colonies, or are under consideration as classical biological control agents. Empirical studies generally corroborate the evolutionary hypothesis that virulence should be relatively low for pathogen species that utilize vertical transmission as one mechanism for maintenance in the host population. Nevertheless, many entomopathogens with significant effects on host populations are vertically as well as horizontally transmitted. In addition to gaining a better understanding of pathogen-host interactions and population dynamics, studies of the host range and specificity of putative biological control agents can benefit by using transmission studies to better predict ecological host specificity from physiological data. Horizontal transmission requires a tightly organized host-pathogen relationship to succeed, but still involves, albeit restricted by host behavior and pathogen dosage, the physiological susceptibility of the nontarget host. Vertical transmission studies can provide increased stringency for determining the ecological host specificity of a species and may be one very accurate predictor of the ability of a pathogen to successfully host-switch when introduced into a na?ve population.     

A review of open-field host range testing to evaluate non-target use by herbivorous biological control candidates

One of the fundamental challenges of pre-release studies in classical biological weed control is to assess and predict the likelihood and consequences of non-target effects. Unless a candidate biological control agent is proven to be monophagous through conventional starvation and host-specificity tests in quarantine, open-field host range studies can be important in predicting the likelihood of non-target effects since they reveal the host selection of herbivores displaying the whole array of pre- and post-alightment behaviours. Over the course of its 53-year history, the purpose and the design of open-field host range studies have changed considerably, with more recent studies clarifying or refining specific questions related to one or a few test plant species and using a set design. We discuss the opportunities and challenges of this approach and suggest that future open-field host range studies should be more hypothesis-driven and apply different experimental designs that facilitate the interpretation of the results.     

Making host specificity testing more efficient: Exploring the use of abridged test plant lists

Testing the specificity of candidate agents is a key component of risk analysis in weed biological control. This step is often time-consuming due to the numerous plant species that need to be tested under quarantine conditions in the invaded country of the weed species. Here, we examined whether an abridged phylogenetically based test list could be used in the weed's native range to quickly screen the host specificity of candidate agents. Ten plant species were used to test the host specificity of a promising candidate for the biological control of Sonchus oleraceus in Australia, the gall midge, Cystiphora sonchi. No-choice and choice tests were carried out in the native Mediterranean range of the midge. The results showed the midge has potential to threaten native Australian species, as those species showed high infestation levels in no-choice tests and produced significantly higher numbers of galls in choice tests. As a result of this approach, C. sonchi was rapidly discarded from the list of agents to be imported into Australian quarantines for further tests. This study demonstrates that testing a few key phylogenetically related species in the native range may save cost and effort in a weed biological control programme.