Predicting the host range of Nystalea ebalea: Secondary plant chemistry and host selection by a surrogate biological control agent of Schinus terebinthifolia

The safety of weed biological control depends upon the selection and utilization of the target weed by the agent while causing minimal harm to non-target species. Selection of weed species by biological control agents is determined by the presence of behavioral cues, generally host secondary plant compounds that elicit oviposition and feeding responses. Non-target species that possess the same behavioral cues as found in the target weed may be at risk of damage by classical biological control agents. Here we conducted host range tests and examined secondary plant compounds of several test plant species. We studied the specialist herbivore Nystalea ebalea (Lepidoptera: Notodontidae) a Neotropical species, present in Florida as a surrogate biological control agent of the weed, Brazilian peppertree Schinus terebinthifolia, invasive in Florida and Hawaii. We found that the larvae had the greatest survival when fed the target weed, the Neotropical species Spondias purpurea, the Florida native species Rhus copallinum, and the ornamental Pistacia chinensis. Reduced survival and general larval performance were found on the native species Metopium toxiferum and Toxicodendron radicans. Both the volatiles and the allergen urushiols were chemically characterized for all species but urushiol diversity and concentration best predicted host range of this herbivore species. These results provide insight into host selection and utilization by one oligophagous Schinus herbivore. Other potential biological control agents may also be sensitive to plants that contain urushiols and if so, they may pose minimal risk to these native species.     

Pre-release evaluation of the efficacy of the leaf-sucking bug Carvalhotingis visenda (Heteroptera: Tingidae) as a biological control agent for cat's claw creeper Macfadyena unguis-cati (Bignoniaceae)

In classical weed biological control, assessing weed response to simulated herbivory is one option to assist in the prioritization of available agents and prediction of their potential efficacy. Previously reported simulated herbivory studies suggested that a specialist herbivore in the leaf-feeding guild is desirable as an effective biological control agent for cat's claw creeper Macfadyena unguis-cati (Bignoniaceae), an environmental weed that is currently a target for biological control. In this study, we tested (i) whether the results from glasshouse-based simulated herbivory can be used to prioritise potential biological control agents by evaluating the impact of a leaf-sucking tingid bug Carvalhotingis visenda (Drake & Hambleton) (Hemiptera: Tingidae) in quarantine; and (ii) the likely effectiveness of low- and high-densities of the leaf-sucking tingid after its release in the field. The results suggest that a single generation of C. visenda has the potential to reduce leaf chlorophyll content significantly, resulting in reduced plant height and leaf biomass. However, the impact of one generation of tingid herbivory on below-ground plant components, including the roots and tuber size and biomass, were not significant. These findings are consistent with results obtained from a simulated herbivory trial, highlighting the potential role of simulated herbivory studies in agent prioritisation.     

Host range tests reveal Paectes longiformis is not a suitable biological control agent for the invasive plant Schinus terebinthifolia

The most critical step during a weed biological control program is determination of a candidate agent’s host range. Despite rigorous protocols and extensive testing, there are still concerns over potential non-target effects following field releases. With the objective to improve risk assessment in biological control, no-choice and choice testing followed by a multiple generation study were conducted on the leaf-defoliator, Paectes longiformis Pogue (Lepidoptera: Euteliidae). This moth is being investigated as a biological control agent of Schinus terebinthifolia Raddi (Sapindales: Anacardiaceae), which is one of the worst invasive plant species in Florida, USA. Results from no-choice testing showed higher larval survival on S. terebinthifolia (48 %) and its close relative Schinus molle L. (47 %), whereas lower survival was obtained on six non-target species (<25 %). When given a choice, P. longiformis females preferred to lay eggs on the target weed, but oviposition also occurred on four non-target species. An improved performance on the native Rhus aromatica Aiton was found when insects were reared exclusively on this non-target species for one or two generations. Results from host range testing suggest that this moth is oligophagous, but has a preference for the target weed. Non-target effects found during multiple generation studies indicate that P. longiformis should not be considered as a biological control agent of S. terebinthifolia.     

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.     

Assessment of the Importance of Similarity in Carbon Source Utilization Profiles between the Biological Control Agent and the Pathogen in Biological Control of Bacterial Speck of Tomato

Bacterial speck of tomato, caused by Pseudomonas syringae pv. tomato, was used to determine whether similarity in carbon source utilization between a preemptive biological control agent and the pathogen was significant in determining the ability of the bacterium to suppress disease. Similarity in carbon source utilization was quantified as the ratio of the number of tomato carbon sources utilized in vitro by the biological control agent to the number of tomato carbon sources utilized in vitro by the target pathogen (the niche overlap index [NOI]). Suppression of the disease was quantified as the percent reduction in disease severity compared to the pathogen-only control when nonpathogenic bacteria were applied to foliage 48 h prior to the pathogen. In the collection of 36 nonpathogenic bacterial strains, there was a significant (P < 0.01), but weak (r² = 0.25), correlation between reduction in disease severity and similarity in carbon source utilization, suggesting that similarity in carbon source use was significant in determining ability to suppress disease. The relationship was investigated further using catabolic mutants of P. syringae strain TLP2, an effective biological control agent of speck. Catabolic mutants exhibited lower levels of similarity (NOI = 0.07 to 0.90) than did wild-type TLP2 (NOI = 0.93). With these catabolic mutants there was a significant (P < 0.01), and stronger (r² = 0.42), correlation between reduction in disease severity and similarity in carbon source utilization. This suggests that similarity in carbon source utilization was a more important component of biological control ability for the catabolic mutants than for the nonpathogenic bacteria. Together, these studies indicate that suppression of bacterial speck of tomato was correlated with nutritional similarity between the pathogenic and nonpathogenic bacteria and suggest that preemptive utilization of carbon sources was probably involved in the biological control of the disease by both the naturally occurring nonpathogenic bacteria and the catabolic mutants.     

Maintenance of a narrow host range by Oxyops vitiosa; a biological control agent of Melaleuca quinquenervia

Host range expansion in insect herbivores is often thought to be mediated by several factors, principal among them are secondary plant metabolites. In weed biological control, the host range of a prospective agent is one of the most important considerations in its implementation. Extensive host testing tests seek to determine the behavioral acceptance and nutritional value of different test plant species to the potential agent. A list of test plants is compiled that comprises species that are close taxonomic relatives of the target weed plus other species of economic or ecologic importance. The host testing of the Melaleuca quinquenervia biological control agent Oxyops vitiosa indicated that larvae would accept and complete development on the Australian target weed M. quinquenervia, two Australian ornamental species, Callistemon citrina, Callistemon viminalis (all Myrtaceae). However, the larvae did not complete development when fed a North American species Myrica cerifera (Myricaceae). The study reported here confirms these results and examines the nutritional and performance differences in O. vitiosa larvae fed leaves of these species. The leaf quality factors, percent moisture, percent nitrogen, toughness, and terpenoid content were related to larval survival, performance and digestive indices. The results indicate that plant quality among the Myrtaceae species was generally similar and correspondingly larval survival, performance and digestive indices differed little when larvae were fed leaves of these species. However, significant differences occurred in the plant quality of the North American M. cerifera compared with the Australian species which had leaves with the lowest percent moisture, lowest leaf toughness, highest percent nitrogen. This species, however, is not a physiological host as none of the neonates survived to pupate. When third instars were switched to M. cerifera from their normal host M. quinquenervia reductions were found in survival, biomass gain, digestive efficiency, and conversion of digested food to insect biomass. The marginal acceptance of this North American native plant in laboratory bioassays appears related to the terpenoid chemistry that has similarities to the taxonomically unrelated host M. quinquenervia. However, the high larval mortality corresponds to several novel terpenoids that are not present in the host. For weed biological control host testing these results indicate that M. cerifera is a poor host for O. vitiosa. Additionally, future test plant lists should include plants with secondary metabolites similar to the target weed as these compounds may constitute behavioral cues that are relevant to these specialized herbivores.     

Review of Pasteuria penetrans: Biology,Ecology, and Biological Control Potential

Pasteuria penetrans is a mycelial, endospore-forming, bacterial parasite that has shown great potential as a biological control agent of root-knot nematodes. Considerable progress has been made during the last 10 years in understanding its biology and importance as an agent capable of effectively suppressing root-knot nematodes in field soil. The objective of this review is to summarize the current knowledge of the biology, ecology, and biological control potential of P. penetrans and other Pasteuria members. Pasteuria spp. are distributed worldwide and have been reported from 323 nematode species belonging to 116 genera of free-living, predatory, plant-parasitic, and entomopathogenic nematodes. Artificial cultivation of P. penetrans has met with limited success; large-scale production of endospores depends on in vivo cultivation. Temperature affects endospore attachment, germination, pathogenesis, and completion of the life cycle in the nematode pseudocoelom. The biological control potential of Pasteuria spp. have been demonstrated on 20 crops; host nematodes include Belonolaimus longicaudatus, Heterodera spp., Meloidogyne spp., and Xiphinema diversicaudatum. Pasteuria penetrans plays an important role in some suppressive soils. The efficacy of the bacterium as a biological control agent has been examined. Approximately 100,000 endospores/g of soil provided immediate control of the peanut root-knot nematode, whereas 1,000 and 5,000 endospores/g of soil each amplified in the host nematode and became suppressive after 3 years.     

Regulation of greenhouse sciarid fly populations using Tetradonema plicans (Nematoda: Mermithoidea)

Tetradonema plicans (Tetradonematidae) satisfied many of the requirements for an efficient biological control agent. It was highly pathogenic and specific to the target hosts, sciarid flies, particularly Bradysia paupera (Lycoriidae). It could be mass-reared, stored as eggs for up to one year at 10°C, and applied to the soil surface as eggs in water. It had high reproductive capacity, each female producing up to 12,000 viable eggs. Since T. plicans was a naturally occurring parasite B. paupera both life cycles could be synchronized.     

Biological control of weeds: an analysis of introductions,rates of establishment and estimates of success,worldwide

The foremost document that comprehensively reports on biological control introductions against weeds—‘Biological control of weeds: a world catalogue of agents and their target weeds’—has been updated and now includes all deliberate releases made through 2012. It includes data on 1555 intentional releases of 468 biological control agent species used against 175 species of target weeds in 48 plant families, in 90 countries. For 55 (31.4%) of the target weed species, only one biocontrol agent was introduced. The largest number of agent species (44) was introduced for the biological control of Lantana camara (Verbenaceae). Three insect orders (Coleoptera, Lepidoptera and Diptera) comprised about 80% of all biocontrol agent species released and releases made. Of the 468 biocontrol agent species introduced, 332 (70.9%) established in at least one instance. Of the 313 species, for which impact could be categorized, 172 (55.0%) caused medium, variable or heavy levels of damage (impacts). Of all releases made through 2012, 982 (63.2%) led to establishment. Forty-two releases were judged too early post-release to categorize impact, leaving 940 releases for which impact analyses were conducted. Similar to agent species, approximately half of the established releases (503 or 53.5%) caused medium, variable or heavy levels of damage on the target weeds, and almost a quarter of releases (225 or 23.9%) caused heavy impact. Across all countries and regions, 65.7% of the weeds targeted for biological control experienced some level of control. These data indicate the value of this practice, on its own, or as a supplement to other methods, in the management of invasive plants.     

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.     

Resource regulation of an invasive tree by a classical biological control agent

The invasive tree Melaleuca quinquenervia experienced substantial declines in growth and reproduction in response to chronic herbivory by the defoliating weevil Oxyops vitiosa. Plants subjected to unrestricted defoliation replaced leaves that were more suitable for feeding by the next generation, a process envisioned by the Resource Regulation Hypothesis which posits that attack by one generation increases the amount of the preferred host resources for the next, resulting in a positive feedback loop for the herbivore. The production of juvenile replacement leaves stimulated additional bouts of oviposition and feeding by O. vitiosa, which ultimately produced positive effects for the herbivore with negative consequences for the plant. The addition of water resources to the plant prolonged the positive feedback loop such that more than twice as many insects were produced on irrigated versus non-irrigated trees. In a more simple, reassembled food web on M. quinquenervia, the lack of biotic constraints like parasitoids may have prevented the earlier termination of the feedback loop and thus increased the impact of the biological control agent on the target. The overall effectiveness of this classical biological control program can be attributed, in part, to the phenomenon of the target plant’s induced susceptible response to a herbivore.     

Ecological host-range of Lilioceris cheni (Coleoptera: Chrysomelidae), a biological control agent of Dioscorea bulbifera

Open-field host-specificity testing assesses the host-range of a biological control agent in a setting that permits the agent to use its full complement of host-seeking behaviors. This form of testing, particularly when it includes a no-choice phase in which the target weed is killed, may provide the most accurate assessment of the ecological host-range of an agent. We conducted a two-phase field host-specificity test with experienced and naïve adults of Lilioceris cheni Gressitt and Kimoto (Coleoptera: Chrysomelidae), a biological control agent of Dioscorea bulbifera L. (Dioscoreales: Dioscoreaceae). We followed field tests with a no-choice laboratory consumption study with the congeneric plant species that received test feeding in the field, and an additional field evaluation of spillover risk. Both experienced and naïve adults strongly preferred D. bulbifera to non-targets in the field. Within 47 h post-release, 90% of the released beetles that remained in the plots were found on D. bulbifera. In the laboratory no-choice test, the beetles consumed significantly more D. bulbifera and survived longer on this plant than the non-targets. All naïve beetles in the Dioscorea sansibarensis and Dioscorea villosa treatments and 75% of naïve beetles on Dioscorea floridana died within 7 d. Potted plants of the native D. floridana experienced minor test feeding in the spillover experiment when surrounded by large populations of L. cheni in the field. At the end of this experiment, L. cheni eggs and/or larvae were present on 83% of D. bulbifera plants but none of the D. floridana plants. We conclude that L. cheni is host-specific to D. bulbifera and does not pose a spillover risk to the native D. floridana.     

Control of Pseudophilothrips ichini (Thysanoptera: Phlaeothripidae) with acephate to exclude a biological control agent of Schinus terebinthifolia

Post-release monitoring of biological control agents to determine impact on the target weed has recently received increased priority. Several methods are available to measure the impact of a biological control agent by manipulating the agent population while measuring fitness of the weed. Brazilian peppertree, Schinus terebinthifolia is one of the most damaging weeds in subtropical areas of Florida and Hawaii. A biological control agent, the thrips, Pseudophilothrips ichini is a sap-feeder that shows high levels of specificity and causes severe distortion of leaf tips of the weed. Thrips populations of this species and a generalist thrips, the red banded thrips Selenothrips rubrocinctus were experimentally manipulated by applications of the systemic insecticide acephate by both foliar applications and by an inserted encapsulated formulation. Foliar applications protected plants against red banded thrips for 29 days and against the biological control thrips, P. ichini for 22 days after treatment. Control with inserts were initially low but was achieved after 60 days and this control continued for 182 days after treatment. Manipulation of these biological control thrips populations with foliar or inserted formulations will assist in the determination of biological control agent impact.     

Impact of herbivory on mile-a-minute weed (Persicaria perfoliata) seed production and viability

The monophagous weevil Rhinoncomimus latipes Korotyaev (Coleoptera: Curculionidae) has been introduced into North America as a biological control agent for the invasive vine mile-a-minute weed, Persicaria perfoliata (L.) H. Gross. This weevil has been shown to reduce the percent cover of mile-a-minute and the number of seed clusters produced, as well as altering the phenology of seed production. However, prior work has not examined the potential impact of the weevil on seed viability and numbers of seeds per cluster. When both adult and larval weevils fed on whole plants in the greenhouse, seed production and fruit maturation were delayed and both the total number of seeds and number of seeds per cluster were reduced. Overall, weevils in greenhouse cages reduced the reproductive potential of P. perfoliata by 35%. When adult weevils were confined on developing seed clusters in the field, the number of seeds produced per cluster, the weight of seeds produced, and seed viability all decreased. Overall, the number of viable seeds per cluster was reduced by 37% in the presence of adult weevils. Taken together, these findings demonstrate that this biological control agent can have important impacts on the reproductive potential of its target weed through mechanisms that have not previously been shown.     

Construction of a Range of Derivatives of the Biological Control Strain Agrobacterium rhizogenes K84: a Study of Factors Involved in Biological Control of Crown Gall Disease

The biological control strain Agrobacterium rhizogenes K84 is an effective agent in the control of Agrobacterium pathogens, the causative agents of crown gall disease. A number of factors are thought to play a role in the control process, including production of the specific agrocins 84 and 434, which differ in the spectra of pathogenic strains that they inhibit in vitro. A range of derivatives of strain K84 has been developed with every combination of the three resident plasmids, pAgK84, pAgK434, and pAtK84b, including a plasmid-free strain. These derivatives produced either both, one, or neither of the characterized agrocins 84 and 434 and were isolated by plasmid curing, conjugation, and Tn5 transposon mutagenesis. The ability of the derivative strains to inhibit gall formation on almond roots was compared to that of the wild-type K84 parent. Treatment with the plasmid-free derivative did not result in a significant level of control of an A. rhizogenes pathogen based on numbers or dry weight of galls formed on injured almond roots. The presence of plasmid pAgK84, pAgK434, or pAtK84b significantly enhanced the biological control efficacy of K84 derivatives, and the highest level of control was observed with strains harboring two or more plasmids. The results observed with strains deficient in agrocin 434 production suggest that this product may play an important role in the biological control of A. rhizogenes pathogens. The involvement of plasmid pAgK84b in biological control has not previously been reported. This study supports the conclusion that multiple factors are involved in the success of strain K84 as a biological control agent.     

Spatial distribution and antifungal interactions of a Bacillus biological control agent on wheat surfaces

The proximity of a biological control agent and its associated anti-microbial metabolites to pathogens on plant surfaces can determine the outcome of disease control. In this study we investigated whether deficiencies in inoculum deposition and localization could explain the inability of the biological control agent Bacillus amyloliquefaciens strain TrigoCor to consistently control Fusarium head blight in the field, despite producing effective and consistent disease control in greenhouse settings. Using epifluorescent stereomicroscopy and confocal laser scanning microscopy, we visualized the coverage of wheat spike surfaces by Bacillus post-application in greenhouse and field environments, and determined that there are large unprotected areas on wheat spikes sprayed with commercial-scale field equipment, as compared to typical greenhouse applications. Additionally, we found that in conditions of low relative humidity, antifungal compounds produced by Bacillus were not able to diffuse across wheat surfaces in biologically relevant amounts, further suggesting that the inadequate coverage of wheat surfaces by Bacillus could be directly limiting disease control. Bacillus cells were easily rinsed off wheat surfaces within 8 h of application, indicating that rainfastness might be an additional limitation of biological control in field settings. Finally, we observed the inhibition of Fusarium graminearum spore germination by TrigoCor inoculum on wheat surfaces, confirming this as a mode of action for TrigoCor biocontrol. Future optimization efforts for biological control agents applied to above-ground plant parts should focus on enhancing the rainfastness, quantity, and spatial coverage of the inoculum on plant surfaces.     

Field observations on the biology and behaviour of Dasiops caustonae Norrbom and McAlpine (Dipt., Lonchaeidae), as a candidate biocontrol agent of Passiflora mollissima in Hawaii

A newly described lonchaeid from Venezuela, Dasiops caustonae Norrbom and McAlpine, has been recommended as a biological control agent of Passiflora mollissima (H.B.K) Bailey, an aggressive exotic weed of Hawaiian rainforests. In this study, elementary biological information was collected in the field in order to determine the appropriateness of using D. caustonae as a biological control agent against this important weed. Field observations suggested that the host range of D. caustonae is limited to species of the subgenus Tacsonia. Female flies appear to mark flowers with an oviposition deterrent pheromone. This is possibly the first report of marking behaviour outside Tephritidae. The geographical range of this species is wide and comparable with that of the target weed. Life‐cycle studies demonstrated that D. caustonae is multivoltine with an estimated life‐cycle length of 3 to 4 months. Survival to adult was limited by premature flower fall, drought and interspecific competition by a bud and flower feeding Lepidoptera, Pyrausta perelegans Hampson. Additional ecological studies and host range testing of D. caustonae are recommended to determine its impact on the target weed and its safety as a biological control agent.     

Multi-Action Planning for Threat Management: A Novel Approach for the Spatial Prioritization of Conservation Actions

Planning for the remediation of multiple threats is crucial to ensure the long term persistence of biodiversity. Limited conservation budgets require prioritizing which management actions to implement and where. Systematic conservation planning traditionally assumes that all the threats in priority sites are abated (fixed prioritization approach). However, abating only the threats affecting the species of conservation concerns may be more cost-effective. This requires prioritizing individual actions independently within the same site (independent prioritization approach), which has received limited attention so far. We developed an action prioritization algorithm that prioritizes multiple alternative actions within the same site. We used simulated annealing to find the combination of actions that remediate threats to species at the minimum cost. Our algorithm also accounts for the importance of selecting actions in sites connected through the river network (i.e., connectivity). We applied our algorithm to prioritize actions to address threats to freshwater fish species in the Mitchell River catchment, northern Australia. We compared how the efficiency of the independent and fixed prioritization approach varied as the importance of connectivity increased. Our independent prioritization approach delivered more efficient solutions than the fixed prioritization approach, particularly when the importance of achieving connectivity was high. By spatially prioritizing the specific actions necessary to remediate the threats affecting the target species, our approach can aid cost-effective habitat restoration and land-use planning. It is also particularly suited to solving resource allocation problems, where consideration of spatial design is important, such as prioritizing conservation efforts for highly mobile species, species facing climate change-driven range shifts, or minimizing the risk of threats spreading across different realms.     

Unraveling the efficient applications of secondary metabolites of various Trichoderma spp.

Recent shift in trends of agricultural practices from application of synthetic fertilizers and pesticides to organic farming has brought into focus the use of microorganisms that carryout analogous function. Trichoderma spp. is one of the most popular genera of fungi commercially available as a plant growth promoting fungus (PGPF) and biological control agent. Exploitation of the diverse nature of secondary metabolites produced by different species of Trichoderma augments their extensive utility in agriculture and related industries. As a result, Trichoderma has achieved significant success as a powerful biocontrol agent at global level. The endorsement of Trichoderma spp. by scientific community is based on the understanding of its mechanisms of action against a large set of fungal, bacterial and in certain cases viral infections. However, it is still an agnostic view that there could be any single major mode of operation, although it is argued that all mechanisms operate simultaneously in a synchronized fashion. The central idea behind this review article is to emphasize the potentiality of applications of target specific secondary metabolites of Trichoderma for controlling phytopathogens as a substitute of commercially available whole organism formulations. With the aim to this point, we have compiled an inclusive list of secondary metabolites produced by different species of Trichoderma and their applications in diverse areas with the major emphasis on agriculture. Outlining the importance and diverse activities of secondary metabolites of Trichoderma besides its relevance to agriculture would generate greater understanding of their other important and beneficial applications apart from target specific biopesticides.