Biocontrol potential of plant growth-promoting rhizobacteria against plant disease and insect pest.

Biological control is a promising approach to enhance pathogen and pest control to ensure high productivity in cash crop production. Therefore, PGPR biofertilizers are very suitable for application in the cultivation of tea plants (Camellia sinensis) and tobacco, but it is rarely reported so far. In this study, production of a consortium of three strains of PGPR were applied to tobacco and tea plants. The results demonstrated that plants treated with PGPR exhibited enhanced resistance against the bacterial pathogen Pseudomonas syringae (PstDC3000). The significant effect in improving the plant's ability to resist pathogen invasion was verified through measurements of oxygen activity, bacterial colony counts, and expression levels of resistance-related genes (NPR1, PR1, JAZ1, POD etc.). Moreover, the application of PGPR in the tea plantation showed significantly reduced population occurrences of tea green leafhoppers (Empoasca onukii Matsuda), tea thrips (Thysanoptera:Thripidae), Aleurocanthus spiniferus (Quaintanca) and alleviated anthracnose disease in tea seedlings. Therefore, PGPR biofertilizers may serve as a viable biological control method to improve tobacco and tea plant yield and quality. Our findings revealed part of the mechanism by which PGPR helped improve plant biostresses resistance, enabling better application in agricultural production.

Exploring behavioural and physiological adaptations in mountain pine beetle in response to elevated ozone concentrations.

Elevated ozone (eO3) concentrations pose a threat to insect populations by potentially altering their behaviour and physiology. This study investigates the effects of eO3 concentrations on the mountain pine beetle which is a major tree-killing species of conifers in northwestern North America. We are particularly interested in understanding the effects of eO3 concentrations on beetle behaviour and physiology and possible transgenerational impacts on bark beetle broods. We conducted O3-enrichment experiments in a controlled laboratory setting using different O3 concentrations (100-200 ppb; projected for 2050-2100) and assessed various beetle responses, including CO2 respiration, mating behaviour, survival probability, locomotion, and attraction behaviour. Transgenerational impacts on the first and second generations were also analyzed by studying brood morphology, mating behaviour, survival, and pheromone production. We found that beetles exposed to eO3 concentrations had shorter oviposition galleries and reduced brood production. Beetle pheromones were also degraded by eO3 exposure. However, exposure to eO3 also prompted various adaptive responses in beetles. Despite reduced respiration, eO3 improved locomotor activity and the olfactory response of beetles. Surprisingly, beetle survival probability was also improved both in the parents and their broods. We also observed transgenerational plasticity in the broods of eO3-exposed parents, suggesting potential stress resistance mechanisms. This was evident by similar mating success, oviposition gallery length, and brood numbers produced in both control and eO3 concentration treatments. This study demonstrates the sensitivity of mountain pine beetles to increased O3 concentrations, contributing crucial insights into the ecological implications of eO3 concentrations on their populations. Overall, the outcome of this study contributes to informed climate change mitigation strategies and adaptive management practices for the development of resilient forests in response to emerging forest insect pests worldwide.

Effective multidimensional treatment identification of different chemical fertilizers: Response of insect dynamics and rice production.

Effective management of fertilizers is essential in influencing the prevalence of insects in rice (Oryza sativa L.) fields. Over two years (2019-20 and 2020-21), an experiment conducted at Bangladesh Rice Research Institute (BRRI), Habiganj, during the boro season aimed to identify the most effective multidimensional treatment (EMT) by testing various combinations of chemical fertilizers and its effect on rice insects. The goal was to optimize rice grain yield while minimizing harmful insect infestation and supporting natural enemies. Eight different chemical fertilizer applications were used as follows: T1 contained a full mix of nitrogen (N), phosphorus (P), potassium (K), and sulfur (S); T2 had PKS but lacked N; T3 had NKS but lacked P; T4 had NPS but lacked K; T5 had NPK but lacked S; T6 had KS but lacked N and P; T7 had PS but lacked N and K; and T8 lacked all four elements - N, P, K, and S. The relationship between the dynamics of harmful insects and natural enemies was highly positively correlated (r = 0.72 to 0.97). In two consecutive growing years, the 2020-21 season exhibited notably higher counts of harmful insects, with Rice Leafroller (RLR) dominating in the booting stage and White Backed Planthopper (WBPH) in mid-tillering, while Green Mirid Bug (GMB) prevailed among natural enemies across both stages, surpassing insect pest counts, notably GMB, Lady bird beetle (LBB), Carabid beetle (CDB), and Staphylinid (STD). However, the yield was notably higher in the 2019-20 growing season despite these pest pressures. Throughout the mid-tillering and booting stages, T1 consistently exhibited the highest average populations of harmful insects and natural enemies, while T7 demonstrated the lowest count of harmful insects, followed by T2 at both growth stages. Additionally, the highest grain yield (GY) was consistently recorded in T1, followed by T5, T6, and T3, with yields of 7.98 t/ha, 7.63 t/ha, 7.38 t/ha, and 7.33 t/ha, respectively. In both stages, beneficial insects prevailed over harmful ones in all fertilizer applications, with significant declines noted in T2 and T7. Factor analysis showed successful selection for EMT in the MGIDI index for all variables except INT and GY during the 2019-20 season, with selection differentials (SD) ranging from -0.10 to 8.29. However, in 2020-21, selection was achieved for all variables with SD ranging from 0.37 to 6.08. According to the MGIDI index, the top-ranked EMTs were identified as T4 and T3 for the 2019-20 period, and T3 and T5 for the 2020-21 period. The EMT shared in both years, T3, proved effective because of its positive impact on enhancing natural enemies throughout both periods (with SD ranging from 4.76 to 8.29 for 2019-20 and 3.03 to 6.08 for 2020-21), and its notable contribution to rice grain yield (SD = 0.37) in 2020-21. This study uniquely integrates EMT to optimize rice grain yield while simultaneously managing harmful insect infestations and supporting natural enemies, addressing a critical need in sustainable rice cultivation. The suggestion is to give preference to fertilizer application T3, which omits P but contains N and K, to improve rice grain yield and boost natural enemies, thereby reducing harmful insect infestation. Moreover, future investigations should concentrate on refining fertilizer blends to strike a harmony between maximizing yield and fostering ecological robustness in rice cultivation.

Comparative metabolomics reveals how the severity of predation by the invasive insect Cydalima perspectalis modulates the metabolism re-orchestration of native Buxus sempervirens.

The recent biological invasion of box tree moth Cydalima perspectalis on Buxus trees has a major impact on European boxwood stands through severe defoliation. This can hinder further regrowth and threaten survival of populations. In a mesocosm approach and controlled larval density over a 2-month period, responses of B. sempervirens essential and specialized metabolites were characterized using metabolomics, combining 1H-NMR and LC-MS/MS approaches. This is the first metabolome depiction of major Buxus responses to boxwood moth invasion. Under severe predation, remaining green leaves accumulate free amino acids (with the noticeable exception of proline). The leaf trans-4-hydroxystachydrine and stachydrine reached 10-13% and 2-3% (DW), while root content was lower but also modulated by predation level. Larval predation promoted triterpenoid and (steroidal) alkaloid synthesis and diversification, while flavonoids did not seem to have a relevant role in Buxus resistance. Our results reveal the concomitant responses of central and specialized metabolism, in relation to severity of predation. They also confirm the potential of metabolic profiling using 1H-NMR and LC-MS to detect re-orchestration of metabolism of native boxwood after severe herbivorous predation by the invasive box-tree moth, and thus their relevance for plant-insect relationships and ecometabolomics.

Interleg coordination in free-walking bug Erthesina fullo (Hemiptera: Pentatomidae).

Insects can adapt their walking patterns to complex and varied environments and retain the ability to walk even after significant changes in their physical attributes, such as amputation. Although the interleg coordination of intact insects has been widely described in previous studies, the adaptive walking patterns in free-walking insects with amputation of 1 or more legs are still unclear. The pentatomid bug Erthesina fullo exhibits a tripod gait, when walking freely on horizontal substrates, like many other insects. In this study, amputations were performed on this species to investigate changes in interleg coordination. The walking parameters were analyzed, such as the locations of touchdown and liftoff, cycle period, walking speed, and head displacement of intact and amputated insects. The results show that E. fullo displays adaptive interleg coordination in response to amputations. With 1 amputated leg, bugs changed to a 3-unit gait, whereas with 2 amputated legs they employed a wave gait. These data are helpful in exploring the motion mode control in walking insects and provide the theoretical basis for the gait control strategy of robots, when leg failure occurs.

Effect of male parental gamma irradiation on host suitability of its F1 progeny of a lepidopteran tropical pest, Spodoptera litura (Fabr.) towards development and virulence of entomopathogenic nematodes, Steinernema thermophilum.

The suitability of F1 progeny insect larvae of the irradiated male parent, Spodoptera litura (Fabr.) for infective juveniles (IJs) of entomopathogenic nematodes (EPN), Steinernema thermophilum was assessed to comprehend the feasibility of combining EPNs with nuclear pest control tactic. As compared to the control, the IJs induced faster host mortality with reduced proliferation in F1 host larvae. IJs derived from F1 host larvae exhibited almost similar proliferation capacity on normal hosts as in control. Further, the molecular basis of EPNs induced mortality in F1 host larvae was evaluated. Dual stress of EPN infection and irradiation induced downregulation of the relative mRNA expression of antimicrobial genes and upregulated expression of antioxidative genes. A pronounced effect of EPNs in association with irradiation stress was apparent on host mortality. Radiation induced sterile F1 insect larvae of S. litura acted as a reasonably suitable host for EPNs and also provided the environment for developing viable EPNs for their potential use as biocontrol agents.

SNMP1 is critical for sensitive detection of the desert locust aromatic courtship inhibition pheromone phenylacetonitrile.

BACKGROUND: Accurate detection of pheromones is crucial for chemical communication and reproduction in insects. In holometabolous flies and moths, the sensory neuron membrane protein 1 (SNMP1) is essential for detecting long-chain aliphatic pheromones by olfactory neurons. However, its function in hemimetabolous insects and its role for detecting pheromones of a different chemical nature remain elusive. Therefore, we investigated the relevance of SNMP1 for pheromone detection in a hemimetabolous insect pest of considerable economic importance, the desert locust Schistocerca gregaria, which moreover employs the aromatic pheromone phenylacetonitrile (PAN) to govern reproductive behaviors. RESULTS: Employing CRISPR/Cas-mediated gene editing, a mutant locust line lacking functional SNMP1 was established. In electroantennography experiments and single sensillum recordings, we found significantly decreased electrical responses to PAN in SNMP1-deficient (SNMP1-/-) locusts. Moreover, calcium imaging in the antennal lobe of the brain revealed a substantially reduced activation of projection neurons in SNMP1-/- individuals upon exposure to PAN, indicating that the diminished antennal responsiveness to PAN in mutants affects pheromone-evoked neuronal activity in the brain. Furthermore, in behavioral experiments, PAN-induced effects on pairing and mate choice were altered in SNMP1-/- locusts. CONCLUSIONS: Our findings emphasize the importance of SNMP1 for chemical communication in a hemimetabolous insect pest. Moreover, they show that SNMP1 plays a crucial role in pheromone detection that goes beyond long-chain aliphatic substances and includes aromatic compounds controlling reproductive behaviors.

Genome-Wide Gene Birth-Death Dynamics Are Associated with Diet Breadth Variation in Lepidoptera.

Comparative analyses of gene birth-death dynamics have the potential to reveal gene families that played an important role in the evolution of morphological, behavioral, or physiological variation. Here, we used whole genomes of 30 species of butterflies and moths to identify gene birth-death dynamics among the Lepidoptera that are associated with specialist or generalist feeding strategies. Our work advances this field using a uniform set of annotated proteins for all genomes, investigating associations while correcting for phylogeny, and assessing all gene families rather than a priori subsets. We discovered that the sizes of several important gene families (e.g. those associated with pesticide resistance, xenobiotic detoxification, and/or protein digestion) are significantly correlated with diet breadth. We also found 22 gene families showing significant shifts in gene birth-death dynamics at the butterfly (Papilionoidea) crown node, the most notable of which was a family of pheromone receptors that underwent a contraction potentially linked with a shift to visual-based mate recognition. Our findings highlight the importance of uniform annotations, phylogenetic corrections, and unbiased gene family analyses in generating a list of candidate genes that warrant further exploration.

Species identification of livefood flightless fly (Torinido-shoujoubae) through DNA barcoding.

Torinido-shoujoubae, as it is called in Japanese, is a flightless Drosophila sp. that is sold commercially in Japan. This Drosophila sp. is often used as feeds for model organisms such as reptiles and spiders. There is no scientific name provided for the fruit fly that is known as Torinido-shoujoubae, as well as any historical background or data behind this species. There has been a previous study that was conducted through morphological characteristics analysis of the body as well as the male copulatory organ and has been estimated as Drosophila hydei. The objective of this study was to determine the species of this unidentified fly known as Torinido-shoujoubae based on a molecular evidence with a DNA barcoding. Samples were purchased from four separate suppliers to examine whether there are any differences between them. COI regions were amplified using PCR and the sequenced results were aligned against two databases, NCBI and BOLD. Torinido-shoujoubae samples provided from all suppliers were confirmed to be D. hydei.

The consequences of heatwaves for the reproductive success and physiology of the wingless sub-Antarctic fly Anatalanta aptera.

Sub-lethal effects of warming temperatures are an important, yet sometimes overlooked impact of climate change that may threaten the long-term survival of numerous species. This, like many other effects of climate change, is especially concerning for cold-adapted ectotherms living in rapidly warming polar regions. This study examines the effects of warmer temperatures on cold-adapted Diptera, using the long-lived sub-Antarctic sphaerocerid fly, Anatalanta aptera, as a focal species. We conducted two experiments to assess heat stress in adult flies, one varying the intensity of the heat stress (daily heating from 4 °C to 8 °C, 20 °C, or 24 °C) and one varying the frequency of heat stress exposure (heating from 4 °C to 12 °C every one, two, or three days) and examined consequences for reproductive success and metabolic responses. We found that more heat stress reduced reproductive output, but not timing of reproduction. Surprisingly, individuals sampled at different times during heat stress exposure were undifferentiable when all metabolite concentrations were analysed with redundancy analysis, however some individual metabolites did exhibit significant differences. Overall, our findings suggest that warmer temperatures in the sub-Antarctic may put this species at greater risk, especially when combined with other concurrent threats from biological invasions.

Metabolomics analysis of physicochemical properties associated with quality deterioration in insect-infested hawthorn berries.

The sliced and dried hawthorn berries are easily infested by insects during storage. This study aimed to determine the effect of insect infestation on the quality of hawthorn berries and assess the change at metabolite level by analyzing physicochemical property and metabolomics profiling. A total of 184 shared differential metabolites were obtained, mainly including flavonoids, fatty acids, carboxylic acids and derivatives, and nitrogenous compounds. Through receiver operating characteristic curve assessment, 9 significant differential markers were screened out to distinguish insect infestation of hawthorn berries. Correlation analysis showed that the color, total organic acids, total phenolics, and total flavonoids were effective indicators for quality evaluation of insect infestation, and uric acid and hippuric acid can serve as biomarkers for the quality deterioration of hawthorn berries during storage. This study demonstrated that insect infestation could decrease the quality of hawthorn berries from macro and micro perspectives.

Harvesting insect pests for animal feed: potential to capture an unexploited resource.

The demand for animal protein grows as the human population increases. Technological and genetic advances in traditional animal agriculture will not produce enough protein to meet future needs without significant innovations such as the use of insects as protein sources. Insect farming is growing insects, whereas insect harvesting is collecting insects from their natural habitats to produce high-quality protein for animal feed or human food. Intensive agricultural environments produce tremendous quantities of pestiferous insects and with the right harvest technologies these insects can be used as a protein supplement in traditional animal daily rations. An avenue to exploit these insects is to use traps such as the United States Department of Agriculture-Biomass Harvest Trap (USDA-BHT) to efficiently attract, harvest, and store insects from naturally abundant agricultural settings. The modular design allows for a low cost, easy to build and fix device that is user friendly and has customizable attractants to target various pest species. Although insect harvesting faces substantial challenges, including insect biomass quantity, seasonal abundance and preservation, food safety, and economic and nutritional evaluation, the potential for utilizing these pests for protein shows tremendous promise. In this forum, insect harvesting is discussed, including its potential, limitations, challenges, and research needs. In addition, the use of a mass trapping device is discussed as a tool to increase the biomass of insects collected from the environment.

Conceptualization, design, and construction of a novel insect mass trapping device: the USDA Biomass Harvest Trap (USDA-BHT).

The use of insects as animal feed has the potential to be a green revolution for animal agriculture as insects are a rich source of high-quality protein. Insect farming must overcome challenges such as product affordability and scalability before it can be widely incorporated as animal feed. An alternative is to harvest insect pests from the environment using mass trapping devices and use them as animal feed. For example, intensive agricultural environments generate large quantities of pestiferous insects and with the right harvest technologies, these insects can be used as a protein supplement in traditional animal daily rations. Most insect trapping devices are limited by the biomass they can collect. In that context, and with the goal of using wild collected insects as animal feed, the United States Department of Agriculture-Biomass Harvest Trap (USDA-BHT) was designed and built. The USDA-BHT is a valuable mass trapping device developed to efficiently attract, harvest, and store flying insects from naturally abundant agricultural settings. The trap offers a modular design with adjustable capabilities, and it is an inexpensive device that can easily be built with commonly available parts and tools. The USDA-BHT is also user-friendly and has customizable attractants to target various pest species.

Infectivity of Plasmodium parasites to Aedes aegypti and Anopheles stephensi mosquitoes maintained on blood-free meals of SkitoSnack.

BACKGROUND: Aedes and Anopheles mosquitoes are responsible for tremendous global health burdens from their transmission of pathogens causing malaria, lymphatic filariasis, dengue, and yellow fever. Innovative vector control strategies will help to reduce the prevalence of these diseases. Mass rearing of mosquitoes for research and support of these strategies presently depends on meals of vertebrate blood, which is subject to acquisition, handling, and storage issues. Various blood-free replacements have been formulated for these mosquitoes, but none of these replacements are in wide use, and little is known about their potential impact on competence of the mosquitoes for Plasmodium infection. METHODS: Colonies of Aedes aegypti and Anopheles stephensi were continuously maintained on a blood-free replacement (SkitoSnack; SS) or bovine blood (BB) and monitored for engorgement and hatch rates. Infections of Ae. aegypti and An. stephensi were assessed with Plasmodium gallinaceum and P. falciparum, respectively. RESULTS: Replicate colonies of mosquitoes were maintained on BB or SS for 10 generations of Ae. aegypti and more than 63 generations of An. stephensi. The odds of engorgement by SS- relative to BB-maintained mosquitoes were higher for both Ae. aegypti (OR = 2.6, 95% CI 1.3-5.2) and An. stephensi (OR 2.7, 95% CI 1.4-5.5), while lower odds of hatching were found for eggs from the SS-maintained mosquitoes of both species (Ae. aegypti OR = 0.40, 95% CI 0.26-0.62; An. stephensi OR = 0.59, 95% CI 0.36-0.96). Oocyst counts were similar for P. gallinaceum infections of Ae. aegypti mosquitoes maintained on SS or BB (mean ratio = [mean on SS]/[mean on BB] = 1.11, 95% CI 0.85-1.49). Similar oocyst counts were also observed from the P. falciparum infections of SS- or BB-maintained An. stephensi (mean ratio = 0.76, 95% CI 0.44-1.37). The average counts of sporozoites/mosquito showed no evidence of reductions in the SS-maintained relative to BB-maintained mosquitoes of both species. CONCLUSIONS: Aedes aegypti and An. stephensi can be reliably maintained on SS over multiple generations and are as competent for Plasmodium infection as mosquitoes maintained on BB. Use of SS alleviates the need to acquire and preserve blood for mosquito husbandry and may support new initiatives in fundamental and applied research, including novel manipulations of midgut microbiota and factors important to the mosquito life cycle and pathogen susceptibility.

Fatty Acid Profile and Thermal Behavior of Fat-Rich Edible Insect Oils Compared to Commonly Consumed Animal and Plant Oils.

This study compared the physicochemical properties of edible insect oils from silkworm (Bombyx mori) pupa (SP), sago palm weevil (Rhynchophorus ferrugineus) larva (PW), and bamboo caterpillar (Omphisa fuscidentalis; BC) to oils from chicken skin (CK), beef back fat (BF), pork back fat (PF), salmon belly (SB), sea bass belly (BB), coconut (C), and peanut (P). The fatty acid profiles and thermal behaviors (crystallization and melting) of the extracted oils were evaluated. PW and BC oils had more saturated fatty acids (SFAs) than CK, PF, SB, BB, and P oils. SP oil had equivalent SFA content to CK and BB oils. Insect oils exhibited similar monounsaturated fatty acid concentrations in all samples, except C oils. PW and BC oils exhibited a higher content of palmitoleic acid than the other oils. SP oils contained polyunsaturated fatty acids similar to those in SB and BB oils, which were higher than those in PW, BC, CK, BF, and PF oils. SP oil also exhibited the highest concentration of α-linolenic acid (C18:3 n-3). Arachidonic acid (0.01-0.02 g/100 g) in all insect oils was lower level compared to CK, BF, PF, SB, and BB oils. SP oil (0.03 g/100 g) exhibited a slightly higher level of eicosapentaenoic acid compared to PW (0.01 g/100 g) and BC (0.01 g/100 g) oils. The insect oils were liquid at ambient temperature, solid below -15°C, and required less energy (△Hm-max) for melting than other samples. This study indicated that insects, particularly SP, could serve as an alternative source of fat to meet its growing demand.

Stable gullies provide a suitable habitat for functional insects and reduce the threat of pests on crops in farmland of Northeast China.

Gullies with lower altitudes compared to the surrounding environment are widely distributed in farmland of the watershed and their numbers are still expanding. However, it is still unclear how these gullies regulate the functional insects in farmland. In this study, land use types combined with the herbaceous plant, herbicide application, soil moisture, topography and climatic factors during crop growth were considered to understand how gullies influence the dynamics of functional insects in farmland from a watershed (240 ha) of Northeast China. The primary findings demonstrate that the richness and abundance of functional insects are generally greatest in gullies, particularly in stable gullies, and decrease in the following order: forest belts, grasslands, and farmlands within the watershed. Notably, the ratios of beneficial insects to pests (BI/Pest) in terms of richness and abundance were lower in gullies before July but reversed after July, in comparison to farmland. Stable gullies exhibited higher BI/Pest abundance and diversity ratios than developing gullies. The richness and abundance of functional insects were higher in the middle sections of gullies compared to their heads and tails. Furthermore, the ratios of BI/Pest were generally lower in farmlands than in any gully position. Functional insect dynamics were mainly determined by season, followed by plant abundance and biomass in the gullies, and rarely by soil moisture in the both watershed and single gullies scales. Generally, the richness and abundance of functional insects in farmland were mainly influenced by gullies, especially influenced by the gully middle position. Insect composition in farmland influenced by stable gullies was stronger than by developing gullies, and stable gullies were more beneficial in reducing the threat of pests to crops in the farmland of the watershed.

Testing for variation in photoperiodic plasticity in a butterfly: Inconsistent effects of circadian genes between geographic scales.

The genetic components of the circadian clock have been implicated as involved in photoperiodic regulation of winter diapause across various insect groups, thereby contributing to adaptation to adverse seasonal conditions. So far, the effects of within-population variation in these genes have not been well explored. Here, we present an experimental test of the effects of within-population variation at two circadian genes, timeless and period, on photoperiodic responses in the butterfly Pararge aegeria. While nonsynonymous candidate SNPs in both of these genes have previously shown to be associated with diapause induction on a between-population level, in the present experiment no such effect was found on a within-population level. In trying to reconcile these results, we examine sequence data, revealing considerable, previously unknown protein-level variation at both timeless and period across Scandinavian populations, including variants unique to the population studied here. Hence, we hypothesize that these variants may counteract the previously observed diapause-averting effect of the candidate SNPs, possibly explaining the difference in results between the experiments. Whatever the cause, these results highlight how the effects of candidate SNPs may sometimes vary across genetic backgrounds, which complicates evolutionary interpretations of geographic patterns of genetic variation.

The use of insect cell line Sf21 for ecotoxicity testing.

Insect cell lines are finding utility in many areas of biology, but their application as an in vitro tool for ecotoxicity testing has been given less attention. Our study aimed to demonstrate the utility and sensitivity of Sf21 cells to commonly used fungicides: Propiconazole and CuSO4, as well as dimethyl sulphoxide (DMSO) an industrial solvent. Sf21 cells were readily cultured from frozen stocks in 3-4 days and showed utility as an invertebrate in vitro acute toxicity test. The data showed the threshold levels of cell survivability against propiconazole and CuSO4. The EC50 values were 135.1 µM and 3.31 mM respectively. The LOAEL (lowest observed adverse effect level) was ≈ 1 µM for propiconazole and ≈ 10 µM for CuSO4. Culturing of Sf21 cells in media containing the solvent DMSO showed that 0.5% DMSO concentration did not effect cell viability. Sf21 cells are sensitive and useful as a robust ecologically relevant screening tool for acute toxicity testing.