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Two-spotted spider mite, Tetranychus urticae Koch, is a ubiquitous and highly polyphagous plant pest with a worldwide distribution. Diapause is essential for its survival in the areas with unfavour...

Dickeya dianthicola (Samson) is an important pathogen causing blackleg disease of potato. Previous work suggested that insects might vector species of Pectobacteriaceae between plants but no conclusive work has confirmed this. Green peach aphids (Myzus persicae Sulzer) and Colorado potato beetles (Leptinotarsa decemlineata Say) are aggressive potato pests and related to known vectors of several species of bacteria other than D. dianthicola. This study sought to determine whether these insects vector D. dianthicola for potato infection. Neither insect species showed olfactory discrimination based on the presence of infection in laboratory tests but beetles were repelled by uninfected foliage treated with 2,3-butanediol, a primary Dickeya metabolite. Beetle recruitment to plants was not affected by their infection status; however, aphids preferred uninfected foliage when conspecifics were present. In the laboratory, neither insect acquired or transmitted D. dianthicola through feeding. In the field, neither insect’s abundance was significantly correlated with disease spread. Overall, this study did not find indications that D. dianthicola is vectored by the tested insect species. Therefore, efforts to limit Dickeya spread should focus on sanitation, water management, and seed screening—not on the control of these insect species. [Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

HighlightsBlack soldier fly larvae can be used to convert agricultural wastes into animal feed.A prototype automated incubation system for producing black soldier fly larvae was designed and tested.The system was successful in growing larvae to a harvestable size.The system retained metabolic heat generated by larval and microbial activity.Abstract. Biological conversion of agricultural wastes into animal feed ingredients using larvae of black soldier fly, (Hermetia illucens) is a promising technology that improves the sustainability of agriculture. We designed and tested a prototype automated incubation system for producing black soldier fly larvae. The system consisted of six 50 L plastic bins enclosed on a ventilated metal rack (178 cm high, 66 cm wide). Water was supplied to maintain a moisture level of about 60% in each bin via soaker hoses connected to sensor-activated solenoid valves. The system was successful in maintaining moisture and temperature suitable for larval development and for growing larvae to harvestable size with minimal labor and energy consumption. Biological activity in the bins generated a considerable amount of metabolic heat, most of which was trapped in the substrate in each bin. This heat may be valuable for rearing black soldier fly larvae in areas with cool climates because this species has low tolerance of cold temperatures but challenging in areas with warm climates. Keywords: Biological conversion, Black soldier fly, Metabolic heat generation, Waste management.

Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), is an oligophagous herbivore that feeds on several plant species in the family Solanaceae. Cannibalism is common in this species and accounts for a significant part of natural field mortality. We investigated effects of feeding on potato (Solanum tuberosum L., preferred host), tomato (Solanum lycopersicum L., non‐preferred host), and pothos [Epipremnum aureum (Linden & Andre) Bunting (Araceae), non‐host] leaves on the incidence of adult Colorado potato beetles predating each other. After 48 h of confinement in laboratory arenas, significantly higher cannibalism was observed in the absence of host vegetation than in the presence of either potato or tomato leaves. After 72 h, more beetles were attacked when kept on tomato leaves than on potato leaves. Being confined on non‐host pothos leaves led to higher cannibalism compared to either species of host plant. However, its incidence was lower compared to the no‐leaf treatment, probably because pothos leaves served as a source of water. Our results suggest that adult cannibalism in the Colorado potato beetle may be triggered by a relatively mild adversity, but its extent increases as conditions deteriorate.

Black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae) can be used for recycling a wide variety of organic wastes. However, these wastes are also suitable for colonisation by pest fly species, including the blow fly Lucilia sericata (Meigen) (Diptera: Calliphoridae), and by pathogenic microorganisms. We tested effects of the presence of black soldier fly larvae (BSFL) on recruitment and retention of blow fly larvae, as well as on composition and dynamics of microbial communities in finfish-based substrates. Substrates inhabited by BSFL contained approximately one-tenth of the L. sericata pupae found in the control substrate even when BSFL were removed prior to the introduction of gravid L. sericata females. Furthermore, almost three-quarters of L. sericata released into substrates previously colonised by BSFL emigrated within 24 h, while no such emigration was detected for the control substrates. Unlike previous studies, bacterial community was heavily dominated by Gram-positive species. Overall bacterial abundance on BSFL substrates declined approximately two-fold compared to the control substrate, while fungal abundance increased. However, surviving microbial communities were still robust and diverse, and continued to be dominated by Gram-positive bacteria. On the other hand, Gram-negative Shigella sonnei, which is a common cause of diarrhoea in humans, was more than 200 times less abundant in BSFL substrates compared to the control throughout the experiment. Repellent and antibiotic properties observed in this study are advantageous for using BSFL in remediation of finfish wastes. However, additional treatment of remaining residue may still be necessary to eliminate all biological contamination.

Black soldier fly larvae, Hermetia illucens (L.), are used to convert organic waste streams into insect-based animal feeds. We tested their ability to retain alpha-linolenic acid (ALA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) from feeding substrates, which has important implications for their use in aquaculture. When supplementing a chicken feed diet with increasing concentrations of salmon oil (0–42%) over an increasing number of days (0–8), the concentrations of the three omega-3 acids in larvae increased significantly. Larval survival and biomass accumulation were not affected. Supplementing a chicken feed diet with increasing concentrations (0–14%) of Tetraselmis chui Butcher (Chlorodendrales: Chlorodendraceae) microalgae paste also significantly increased ALA and EPA contents of the harvested larvae. However, microalgae also decreased survival, harvested biomass, and individual growth of larvae feeding on the diet with the highest supplement concentration (14%). DHA was not detected in any microalgae diet or subsequent larval tissue samples. All three omega-3 polyunsaturated fatty acids tested in this study were accumulated in dose-dependent manner, with quadratic, and occasionally linear, equations providing the best description of the observed relationships. There were significant negative correlations between several fatty acids, indicating that they may replace one another in living larvae. Our findings confirm that black soldier fly larvae can retain ingested fatty acids and change fatty acid profiles in their tissues accordingly. However, optimizing nutrient content of harvestable larvae is likely to be more complicated than simply enriching their diets with omega-3 fatty acids.

Potato virus Y (PVY) is a non-persistent virus that is transmitted by many aphid species and causes significant damage to potato production. We constructed a spatially-explicit model simulating PVY spread in a potato field and used it to investigate possible effects of transmission efficiency, initial inoculum levels, vector behavior, vector abundance, and timing of peak vector activity on PVY incidence at the end of a simulated growing season. Lower PVY incidence in planted seed resulted in lower virus infection at the end of the season. However, when populations of efficient PVY vectors were high, significant PVY spread occurred even when initial virus inoculum was low. Non-colonizing aphids were more important for PVY spread compared to colonizing aphids, particularly at high densities. An early-season peak in the numbers of noncolonizing aphids resulted in the highest number of infected plants in the end of the season, while mid- and late-season peaks caused relatively little virus spread. Our results highlight the importance of integrating different techniques to prevent the number of PVY-infected plants from exceeding economically acceptable levels instead of trying to control aphids within potato fields. Such management plans should be implemented very early in a growing season.

Colorado potato beetle (CPB,Leptinotarsa decemlineata) is a major pest of potato and other solanaceous vegetables in the Northern Hemisphere. The insect feeds on leaves and can completely defoliate crops. Because of the repeated use of single insecticide classes without rotating active ingredients, many chemicals are no longer effective in controlling CPB. Ledprona is a sprayable double-stranded RNA biopesticide with a new mode of action that triggers the RNA interference pathway. Laboratory assays with second instar larvae fed Ledprona showed a dose–response where 25×10−6g/L of dsPSMB5 caused 90% mortality after 6days of initial exposure. We also showed that exposure to Ledprona for 6h caused larval mortality and decreased target messenger RNA (mRNA) expression. Decrease in PSMB5 protein levels was observed after 48h of larval exposure to Ledprona. Both PSMB5 mRNA and protein levels did not recover over time. Ledprona efficacy was demonstrated in a whole plant greenhouse trial and performed similarly to spinosad. Ledprona, currently pending registration at EPA, represents a new biopesticide class integrated pest management and insecticide resistance management programs directed against CPB.

Arthropod Glutathione S-transferases (GSTs) constitute a large family of multifunctional enzymes that are mainly associated with xenobiotic or stress adaptation. GST-mediated xenobiotic adaptation takes place through direct metabolism or sequestration of xenobiotics, and/or indirectly by providing protection against oxidative stress induced by xenobiotic exposure. To date, the roles of GSTs in xenobiotic adaptation in the Colorado potato beetle (CPB), a notorious agricultural pest of plants within Solanaceae, have not been well studied. Here, we functionally expressed and characterized an unclassified-class GST, LdGSTu1. The three-dimensional structure of the LdGSTu1 was solved with a resolution up to 1.8 Å by X-ray crystallography. The signature motif VSDGPPSL was identified in the “G-site”, and it contains the catalytically active residue Ser14. Recombinant LdGSTu1 was used to determine enzyme activity and kinetic parameters using 1-chloro-2, 4-dinitrobenzene (CDNB), GSH, p-nitrophenyl acetate (PNA) as substrates. The enzyme kinetic parameters and enzyme-substrate interaction studies demonstrated that LdGSTu1 could catalyze the conjugation of GSH to both CDNB and PNA, with a higher turnover number for CDNB than PNA. The LdGSTu1 enzyme inhibition assays demonstrated that the enzymatic conjugation of GSH to CDNB was inhibited by multiple pesticides, suggesting a potential function of LdGSTu1 in xenobiotic adaptation.

Arthropod Glutathione S-transferases (GSTs) constitute a large family of multifunctional enzymes that are mainly associated with xenobiotic or stress adaptation. GST-mediated xenobiotic adaptation is through direct metabolism or sequestration of xenobiotics, and/or indirectly by providing protection against oxidative stress induced by xenobiotic exposure. To date, the roles of GSTs in xenobiotic adaptation in the Colorado potato beetle (CPB), a notorious agriculture pest of plants within Solanaceae have not been well studied. Here, we functionally expressed and characterized an unclassified-class GST, LdGSTu1. The three-dimensional structure of the LdGSTu1 was solved with a resolution up to 1.8 Å by x-ray crystallography. Recombinant LdGSTu1 was used to determine enzyme activity and kinetic parameters using 1-chloro-2,4-dinitrobenzene (CDNB), GSH, p-nitrophenyl acetate (PNA) as substrates. The enzyme kinetic parameters and enzyme-substrate interaction studies demonstrated that LdGSTu1 could catalyze the conjugation of GSH to both CDNB and PNA, with a higher turnover number for CDNB than PNA. The LdGSTu1 enzyme inhibition assays demonstrated that the enzymatic conjugation of GSH to CDNB could be inhibited by multiple pesticides, suggesting a potential function of LdGSTu1 in xenobiotic adaptation.

Two-spotted spider mite, Tetranychus urticae Koch (Tetranychidae: Acariformes), is one of the most important agricultural pests in the world. Their populations have a tendency of rapidly developing resistance to acaricides, making it necessary to have a variety of active ingredients for sustainable chemical control of this pest. We investigated acaricidal properties of a relatively new insecticide spirotetramat using its commercial formulation, Movento Energy. Spirotetramat applied at concentrations equivalent to the field rates of 78 and 90 g ai/ha had a strong negative effect on the survival of the treated T. urticae, with ca. 95% of females and ca. 65% of males dying after the treatment. Spirotetramat appeared to interfere with female reproductive system. Many of the dead treated females had eggs stuck in the oviduct and protruding from their bodies. Surviving treated females did not lay eggs. Furthermore, untreated females that mated with treated males did not produce female offspring and displayed the symptoms of spirotetramat poisoning. Toxic effects continued manifesting themselves after female mites were transferred from treated onto untreated plant culture. Contrary to previous studies, contact toxicity was also detected. None of the treated immature stages survived to adulthood. Based on these results, spirotetramat may be a good option for integrated pest management in crops that are simultaneously affected by sucking insects and spider mites and in pesticide rotation sequences that are a part of integrated resistance management programs.

Insecticidal double-stranded RNAs (dsRNAs) silence expression of vital genes by activating the RNA interference (RNAi) mechanism in insect cells. Despite high commercial interest in insecticidal dsRNA, information on resistance to dsRNA is scarce, particularly for dsRNA products with non-transgenic delivery (ex. foliar/topical application) nearing regulatory review. We report the development of the CEAS 300 population of Colorado potato beetle (Leptinotarsa decemlineata Say) (Coleoptera: Chrysomelidae) with > 11,100-fold resistance to a dsRNA targeting the V-ATPase subunit A gene after nine episodes of selection using non-transgenic delivery by foliar coating. Resistance was associated with lack of target gene down-regulation in CEAS 300 larvae and cross-resistance to another dsRNA target (COPI β; Coatomer subunit beta). In contrast, CEAS 300 larvae showed very low (~ 4-fold) reduced susceptibility to the Cry3Aa insecticidal protein from Bacillus thuringiensis. Resistance to dsRNA in CEAS 300 is transmitted as an autosomal recessive trait and is polygenic. These data represent the first documented case of resistance in an insect pest with high pesticide resistance potential using dsRNA delivered through non-transgenic techniques. Information on the genetics of resistance and availability of dsRNA-resistant L. decemlineata guide the design of resistance management tools and allow research to identify resistance alleles and estimate resistance risks.

The Colorado potato beetle (CPB) is a prominent insect pest of potatoes, tomatoes and eggplants all over the world, however, the management of CPB remains a challenging task for more than one hundred years. We have successfully developed bacteria-expressed dsRNA-mediated feeding RNA interference (RNAi) approach in our previous study. A critical step towards field management of CPB via feeding RNAi is to identify effective and environmentally safe target genes. NADPH-Cytochrome P450 reductase (CPR) plays a central role in cytochrome P450 action. The full length Leptinotarsa decemlineata CPR (LdCPR) cDNA was isolated from an imidacloprid resistant population. The LdCPR gene was ubiquitously expressed in all stages tested but showed an increase in expression during the early stage of embryonic development. The bacteria-expressed dsRNA-mediated feeding RNAi of LdCPR in adults caused systemic knock down expression of the gene coding for LdCPR in both adults and their eggs. Suppression of LdCPR expression increased susceptibility of imidacloprid in resistant beetles, as well as a significant decrease of fecundity in female beetles (29% less eggs/day) and the hatching rate (47%) of their eggs. These data suggest that LdCPR plays important roles in insecticide detoxification and biosynthetic pathways of endogenous compounds and may serve as an essential target to control CPB.HIGHLIGHTSHigh expression of LdCPR was observed in the egg stage.Silencing of LdCPR reduced the CPR enzymatic activities.LdCPR knockdown increased imidacloprid susceptibility.LdCPR knockdown decreased the fecundity and enhanced embryonic lethality.

Eastern New York State is frequently the site of Colorado potato beetle (Leptinotarsa decemlineata, Say) populations with the highest observed levels of insecticide resistance to a range of active ingredients. The dominance of a resistant phenotype will affect its rate of increase and the potential for management. On organic farms on Long Island, L. decemlineata evolved high levels of resistance to spinosad in a short period of time and that resistance has spread across the eastern part of the Island. Resistance has also emerged in other parts of the country as well. To clarify the level of dominance or recessiveness of spinosad resistance in different parts of the United States and how resistance differs in separate beetle populations, we sampled in 2010 beetle populations from Maine, Michigan, and Long Island. In addition, a highly resistant Long Island population was assessed in 2012. All populations were hybridized with a laboratory-susceptible strain to determine dominance. None of the populations sampled in 2010 were significantly different from additive resistance, but the Long Island population sampled in 2012 was not significantly different from fully recessive. Recessive inheritance of high-level resistance may help manage its increase.

Black soldier flies, Hermetia illucens (L.), consume decaying organic materials at the larval stage and can be used for recycling a variety of biogenic wastes into value-added products. Black soldier flies are normally found in subtropical and warm temperate regions. Cold temperatures may prevent their establishment in colder areas, thus alleviating a concern of their becoming an invasive species. Potentially, cold temperatures can also be used to manipulate the rate of black soldier fly development, which may be needed for timing certain life stages for mass-production needs. In the present study, immature black soldier flies were highly susceptible to freezing. Their survivorship decreased as time spent at −12°C increased from 10 to 60 min. Only ca. 2% of eggs, 80% of third instars and >90% of fifth instars were still alive following 72 h of exposure. Chilling fifth instars resulted in smaller adults but freezing them for 48 h resulted in bigger adults. Based on these results, black soldier fly is unlikely to establish in areas with long periods of subfreezing winter temperatures. Low temperatures may be used to manipulate development of the late instars, but at a cost of higher mortality.

Variability is an important characteristic of population dynamics, but the length of the time series required to estimate population variability is poorly understood. To this end, population variability of Macrosiphum euphorbiae (Thomas), Myzus persicae (Sulzer), and Aphis nasturtii (Kaltenbach) (Hemiptera: Aphididae) was investigated. Population variability (measured as PV, a proportion between 0 and 1) was estimated for time series of 3–62 years, giving replicate estimates for time series of 3–20 years that were normally distributed. Mean values for PV were more uniform for a time series of 12 years or longer than for shorter ones. The standard deviation of PV declined to a minimum at 12–15 years, as the length of the time series increased. Discrimination of estimates of PV was reliable for 15-year time series and longer, but not necessarily for shorter ones. Although M. euphorbiae had a relatively low PV, the coefficient of variation of that PV (12.5), was higher than for the other two species (3.5, 4.5). For robust estimates of PV, a time series of 15 years is recommended, because it minimises the standard deviation of PV, and discriminates values of PV that differ by 0.06 on a 0–1 scale.

Agricultural systems are often simultaneously impacted by multiple stressors. In our study, we investigated the effects of two commonly occurring viruses of potato, Potato virus Y (PVY) and Potato leafroll virus (PLRV) on host plant selection and utilization across various spatial scales by the Colorado potato beetle. Adult beetles selected PLRV-infected foliage in the cage choice trails, and larvae that fed exclusively on PLRV-infected foliage developed into larger adults. PVY-infected foliage was avoided in the cage trials and had no effect on the weight of developing adults. In field trials, preference and utilization of potato plants was more dependent on plant size than on virus infection. These results demonstrate that while virus-infected plants can cause physiological changes that influence beetle preference, beetle abundance and/or distribution is driven by a more complex array of factors at the field level. Additionally, this study demonstrates the importance of investigating the applicability of laboratory findings at the field level.

Microalgae feed production is a major cost in bivalve aquaculture. Its efficiency is increased by scaling it up under automated control of environmental conditions. The initial cost of commercially available systems can be prohibitive. Therefore, a large volume full-scale photobioreactor built of low-cost, readily available materials was developed and tested. A cone-bottom, polyethylene tank was internally illuminated with submersible fluorescent lamps and equipped with a monitoring and control system that measured temperature, pH and optical density of the microalgal culture. Injection of CO2 was automated to maintain pH within a target range. System performance was evaluated by growing four batch cultures of Tetraselmis chuii. Temperature inside the photobioreactor was 29.5 ± 2.38 °C (mean ± SD, range 21−35 °C) and pH was 7.56 ± 0.87 (mean ± SD, range 5.29–8.97). Optimal harvest time was after 300 h (12.5 days) of growth, yielding 1700 L of microalgae at a density of 2500 cells/ μl (1200 cells/ μl/ m2 illuminated surface area). With 24 h illumination, the fluorescent lights, air pump and monitoring/ control device consumed 14.9, 1.9 and 0.1kwh/ day, respectively. Assuming the microalgae are harvested at their optimal density after 12.5 days, each batch would consume 211kwh or 0.124kwh/ liter. The photobioreactor described provides an economical option for growing large amounts of microalgae for aquaculture feed and other purposes.

Cannibalism, or intraspecific predation, can play a major role in changing individual fitness and population processes. In insects, cannibalism frequently occurs across life stages, with cannibals consuming a smaller or more vulnerable stage. Predation of adult insects on one another is considered to be uncommon. We investigated adult cannibalism in the Colorado potato beetle, Leptinotarsa decemlineata (Say), which is an oligophagous herbivore specializing on plants in family Solanaceae, and an important agricultural pest. Under laboratory conditions, starvation and crowding encouraged teneral adults to feed upon each other, which reduced their weight loss during the period of starvation. However, pupae were attacked and consumed before adults. Injured beetles had a higher probability of being cannibalized than intact beetles. Males were more frequently attacked than females, but that appeared to be a function of their smaller size rather than other gender-specific traits. Cannibalizing eggs at a larval stage did not affect beetle propensity to cannibalize adults at an adult stage. When given a choice between conspecific adults and mealworms, the beetles preferred to eat conspecifics. Cannibalistic behavior, including adult cannibalism, could be important for population persistence in this species.

The Colorado potato beetle is an important pest of solanaceous plants in the Northern Hemisphere. Better understanding of its physiological responses to temperature stress and their interactions with still-prevalent chemical control has important implications for the management of this insect. We measured mortality and expression of the Hsp70 heat shock proteins in the Colorado potato beetle larvae exposed to sublethal concentration of the commonly used insecticide imidacloprid, and to supraoptimal temperatures. Both turned out to be significant stress factors, although induction of Hsp70 by imidacloprid observed in the present study was low compared to its induction by the heat. The two factors also interacted with each other. At an extreme temperature of 43 °C, exposure to a sublethal dose of imidacloprid resulted in a significant rise in larval mortality, which was not observed at an optimal temperature of 25 °C. Heat-stressed larvae also failed to respond to imidacloprid by producing more Hsp70. These findings suggest that when field rates of insecticides become insufficient for killing the exposed beetles under optimal temperature conditions due to the evolution of resistance in beetle populations, they may still reduce the probability of resistant beetles surviving the heat shock created by using propane flamers as a rescue treatment.

Mineral oil is a product used to reduce Potato Virus Y transmission in potato fields. However, there is little information available about other effects that oil may have on insect pests of potato. To better understand how mineral oil affects potato pests, we performed a series of experiments testing the effects of oil on mortality, behavior, and development of potato aphids, Macrosiphum euphorbiae (Thomas) (Hemiptera: Aphididae), green peach aphids, Myzus persicae (Sulzer) (Hemiptera: Aphididae), and Colorado potato beetles, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae). All three species showed negative behavioral responses to oil-treated potato foliage. Oil treatment also increased aphid mortality. Colorado potato beetle mortality was not affected, but developing on oil-treated potato plants resulted in prolonged development and smaller adults. Additionally, oil acted synergistically with the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Clavicipitaceae); Colorado potato beetle larvae were killed more rapidly when sprayed with both products compared with when sprayed with B. bassiana alone. Based on these results, mineral oil has the potential for expanded use in potato IPM programs.

Neonicotinoid insecticides have been the most common management tool for Colorado potato beetle, Leptinotarsa decemlineata (Say), infestations in cultivated potato for nearly 20 yr. The relative ease of applying neonicotinoids at planting coupled with inexpensive, generic neonicotinoid formulations has reduced the incentive for potato growers to transition from these products to other mode of action (MoA) groups for early-season L. decemlineata control. Continuous use of neonicotinoids has resulted in resistant L. decemlineata populations in some production areas of the eastern United States. Continued reliance on neonicotinoids will accelerate L. decemlineata resistance development and result in additional insecticide inputs to manage these populations. Resistance management recommendations for L. decemlineata have focused on rotation of insecticides within the growing season. Growers using at-plant neonicotinoids for early-season L. decemlineata control are encouraged to rotate MoAs for later generations to delay resistance development. Although this short-term insecticide rotation has likely prolonged the utility of neonicotinoid insecticides, reducing reliance on a single MoA soil application at planting will improve the longevity of newer, more reduced-risk alternatives. The objectives of this article are twofold: 1) to provide a review of the current status of L. decemlineata neonicotinoid resistance, and 2) to propose long-term resistance management strategies that arrange reduced-risk MoA groups into several, multiyear sequences that will maximize L. decemlineata control and reduce the probability for resistance development. This recommendation maintains practical and economical approaches for L. decemlineata control, but limits reliance on any single MoA group to minimize selection pressure for resistance development. Wisconsin Potato Industry Board, Distinguished Graduate Student Fellowship; R. Keith Chapman and Jeffrey A. Wyman-Wisconsin Distinguished Graduate Fellowship in Vegetable Entomology; National Potato Council-State Cooperative Potato Research Program [FY09-13] We thank cooperating growers for generously providing insecticide application records and allowing us to conduct research on their farms. We also thank the research programs of Jeffrey A. Wyman and R. Keith Chapman for generating L. decemlineata screening data used to determine sustainability improvements of insecticide tools in Wisconsin. We thank the Wisconsin Potato and Vegetable Growers Association (WPVGA) and the New York Empire State Potato Growers Association for continued support of our research efforts. Funding was provided by Wisconsin Potato Industry Board, Distinguished Graduate Student Fellowship 2009 & 2011, R. Keith Chapman and Jeffrey A. Wyman-Wisconsin Distinguished Graduate Fellowship in Vegetable Entomology 2010, and National Potato Council-State Cooperative Potato Research Program FY09-13.

The Colorado potato beetle, Leptinotarsa decemlineata (Say) is a very serious pest of potatoes which is highly mobile and capable of rapid evolution of resistance to chemical control. Insect movement, resulting in gene flow between resistant and susceptible populations, is considered to be an important factor affecting the development and spread of insecticide resistance. We investigated the movement of adult Colorado potato beetles by flight and by walking following the treatment with a sublethal dose of imidacloprid in the laboratory. Imidacloprid had a pronounced negative effect on beetle mobility. The proportion of beetles flying and walking, as well as the number and duration of performed flights, were significantly decreased for the treated beetles. Since local selection followed by long-distance dispersal have been reported to lead to serious area-wide problems with the insecticide-resistant Colorado potato beetle, long-term suppression of flight activity recorded in our study suggests that imidacloprid applications may reduce outflow of resistant alleles.

Phosphite is a general term used to describe the salts of phosphorous acid H 3 PO 3 . It is effective in suppressing a number of plant diseases caused by oomycetes and has been shown to reduce populations of several insect species. We investigated the effects of phosphite on the Colorado potato beetles in the field and laboratory. Beetle numbers and defoliation on phosphite-treated plots were lower compared to the control plots during one out of two years of the study. No phosphite effects were detected in the field during the second year of the study. However, larval mortality was significantly higher the second year in the laboratory when larvae were fed on potato foliage excised from the potato plants treated with phosphite in the field. Laboratory tests with excised leaves dipped in a solution of phosphite revealed lower beetle survivorship and prolonged development on the treated foliage. Because of its dual properties as a fungicide and an insecticide, as well as its low toxicity to vertebrates, phosphite is a potentially good fit for integrated pest management programs.

Colorado potato beetle, Leptinotarsa decemlineata (Say), is a major pest of potatoes in North America, Europe, and parts of Asia. It adapts quickly to adverse environments, thereby limiting the longevity of control strategies. Better understanding of variability among geographically isolated populations of this insect might create an opportunity to customize control techniques for local conditions. We investigated insecticide resistance, heat tolerance, and growth potential in six beetle populations collected from different locations in the United States. Significant differences were detected in insecticide resistance, egg mass size, and egg hatchability among the tested populations. Large egg mass size appeared to be offset by lower egg hatchability. We also observed a female-biased sex ratio at least in one of the strains. Population growth rates were different among the strains, but this parameter was not correlated to insecticide resistance. This suggests that using allopatric populations in comparative studies of insecticide resistance might result in confusing effects of geographic isolation with the effects of selection toward resistance.

Aims Integrating multiple soil and disease manage- ment practices may improve crop productivity and disease control, but potential interactions and limita- tions need to be determined. Methods Three different potential disease-suppressive management practices, including a Brassica napus (rapeseed) green manure rotation crop, conifer-based compost amendment, and three biological control or- ganisms (Trichoderma virens, Bacillus subtilis, and Rhizoctonia solani hypovirulent isolate Rhs1A1) were evaluated alone and in combination at sites with both organic and conventional management histories for their effects on soilborne diseases and tuber yield. Results Rapeseed rotation reduced all observed soil- borne diseases (stem canker, black scurf, common scab, and silver scurf) by 10 to 52 % in at least one year at both sites. Compost amendment had variable effects on tuber diseases, but consistently increased yield (by 9 to 15 %) at both sites. Biocontrol effects on disease varied, though Rhs1A1 decreased black scurf at the conventional site and T. virens reduced multiple diseases at the organic site in at least one year. Combining rapeseed rotation with compost amend- ment both reduced disease and increased yield, where- as biocontrol additions produced only marginal addi- tive effects. Conclusions Use of these treatments alone, and in combination, can be effective at reducing disease and increasing yield under both conventional and organic production practices.

Seasonal patterns of abundance and population variability were determined for Macrosiphum euphorbiae (Thomas), Myzus persicae (Sulzer), and Aphis nasturtii (Kaltenbach) (Hemiptera: Aphididae) in potato plots from weekly samples for 28 years. All species showed a single annual peak, but arrived and reached peak abundance at different times. Population variability (PV, a proportion between 0 and 1) for the week of peak abundance was close to that of other sample weeks and mean seasonal abundance. Based on mid-season abundance, PV of 0.76 for M. persicae differed significantly from 0.80 for A. nasturtii, as well as from 0.59 for M. euphorbiae. A weekly time scale for abundance, initiated at an early stage of plant growth, produced slightly different estimates of PV early and late in the season than a scale centred on peak abundance for each species. PV at the time of invasion differed from estimates for the rest of the summer. The annual abundance used to estimate PV was best determined in the context of aphid life history. Nevertheless, PV provided a robust and precise metric for comparing population variability among the three species, regardless of their seasonal patterns of abundance.

Cultural practices such as organic amendments, rotations, and use of biological control organisms are regularly investigated for their effects on controlling plant diseases but their effects on soil microbial populations are often unexplored. In this study, three different sustainable disease management practices, use of compost amendment, biocontrol organisms, and a potentially disease-suppressive rotation, were established in potato field trials at two sites under different management regimes and histories, and evaluated over three potato cropping seasons for their effects on soil microbial communities. Specific management factors assessed included the presence or absence of a conifer-based compost amendment, addition of one of three different biocontrol organisms (Trichoderma virens, Bacillus subtilis, and Rhizoctonia solani isolate Rhs1A1), and a Brassica napus (rapeseed) green manure rotation crop preceding potato, and treatments were assessed in all factorial combinations. The two farm sites represented organic and conventional potato production practices in Aroostook County, Maine. Compost amendment and rapeseed rotation had the greatest impacts on soil microbial communities, with both treatments increasing total populations of culturable bacteria at both sites over the course of the study, as well as causing detectable shifts in soil microbial community characteristics as determined by sole carbon-source substrate utilization and fatty acid methyl ester (FAME) profiles. Compost amendment generally led to increased utilization of complex substrates and increased levels of Gram-positive bacteria and fungi, and compost effects were more pronounced at the conventional site. Rapeseed rotation often resulted in somewhat different effects at the two different sites. Consistent overall effects were observed with the biocontrol amendments Rhs1A1 and T. virens, including increased microbial activity and bacterial populations. Combined effects of multiple treatments were greater than those of individual treatments and were generally additive. These results indicate that each treatment factor had significant and specific effects on soil microbial communities, and that combined effects tended to be complementary, suggesting the potential of combining multiple compatible management practices and their associated changes in soil microbial communities.

Abundance, persistence, and variability of populations of Macrosiphum euphorbiae (Thomas), Myzus persicae (Sulzer), and Aphis nasturtii Kaltenbach (Hemiptera: Aphididae) in potato plots for intervals of 58 years (n = 1), 29 years (n = 2), 19–20 years (n = 3), and 9–10 years (n = 6) were compared. The abundance of M. euphorbiae showed no trend among decades and varied 2.4-fold, whereas that of M. persicae and A. nasturtii declined and showed 54-fold and 3700-fold variation, respectively. All three aphid species persisted through the first five decades and M. euphorbiae also persisted through the sixth (last) decade, but M. persicae and A. nasturtii failed to persist for 1 and 3 years of the last decade, respectively. Population variability (a proportion between 0 and 1) measured over a 58-year interval was high: 0.585 for M. euphorbiae, 0.771 for M. persicae, and 0.830 for A. nasturtii. During the first three but not the last three decades, population variability increased with sampling interval, owing to dramatic declines in abundance for M. persicae and A. nasturtii and one stable decade for M. euphorbiae, but no evidence of a more-time — more-variation effect was detected. Persistence was not related to population variability, but declined with abundance. Populations did not reach equilibrium, because of declining abundance for M. persicae and A. nasturtii and changes in population variability from decade to decade for M. euphorbiae. Populations of M. persicae and A. nasturtii from this crop monoculture were less stable than previously studied natural populations of a native aphid species. In contrast, the population of M. euphorbiae, a native species, had variability in a potato crop similar to that of the previously studied native species. The high population variability of M. persicae and A. nasturtii may be associated with their status as introduced species. The dynamic and species-specific characteristics of population variability require that interspecific comparisons be considered cautiously.

Novaluron is a chitin synthesis inhibitor (a benzoylphenyl urea) that kills larval stages and causes reversible cessation of laying viable eggs in adults of the Colorado potato beetle, Leptinotarsa decemlineata (Say). Previous studies also suggest that it has ovicidal properties, but not on all tested egg masses. We investigated if egg maturity at the time of exposure affects its susceptibility to novaluron. Novaluron application significantly reduced the hatch of eggs that were 0–24 h old at the moment of treatment, but had no effect on the eggs that were 96–120 h old. Novaluron also interfered with the development of the hatched larvae regardless of the egg age at the time of treatment, probably due to chorion feeding by neonates. Ovicidal properties of novaluron may contribute to Colorado potato beetle control, but their effect will be limited to newly laid eggs. If that window of susceptibility is missed, hatching larvae will be exposed to a fairly old novaluron residue that might be weathered down to sublethal levels. This supports the current recommendations to apply novaluron as a larvicide.

The acceptability of four different aphid species Macrosiphum albifrons (Essig), Macrosiphum euphorbiae (Thomas), Macrosiphum pseudorosae Patch, and Myzus persicae (Sulzer) (Hemiptera: Aphididae), as prey for four lady beetle species, one native species Coccinella trifasciata L, and three non-native Coccinella septempunctata L, Harmonia axyridis Pallas, Propylea quatuordecimpunctata L (Coleoptera: Coccinellidae) were tested in the laboratory. The relative field abundance of adults of the same lady beetle species on host vegetation, Lupinus polyphyllus Lindley (Fabales: Fabaceae), Solanum tuberosum L (Solanales: Solanaceae), and Rosa multiflora Thunberg (Rosales: Rosaceae), both with and without aphids present was also observed. In the laboratory, H. axyridis generally consumed the most aphids, while P. quatuordecimpunctata consumed the fewest. The exception was P. quatuordecimpunctata, which consumed a greater number of M. albifrons nymphs, and C. trifasciata, which consumed a greater number of M. albifrons nymphs and adults, compared with the other two beetle species. Lady beetles consumed fewer M. albifrons compared with the other three aphid species, likely because of deterrent compounds sequestered by this species from its host plant. In the field, P. quatuordecimpunctata was the most abundant species found on L. polyphyllus and S. tuberosum.

Response to odours plays an important role in resource location by natural enemies, particularly by parasitoid wasps. While a considerable research effort has been dedicated to studying the effects of insecticide intoxication on natural enemy search behaviour, it is yet unknown if the odours themselves interfere with distant chemoreception. We investigated this issue using the food-searching behaviour of Microplitis croceipes (Cresson)(Hymenoptera: Braconidae) in laboratory arenas as a model system. Odours of imidacloprid (Genesis®), spinosad (Entrust®), esfenvalerate (Asana®), methamidophos (Monitor®), and vanilla were tested for their ability to interfere with wasp response to the odour of honey. The wasps did not contact the chemicals. Honey odour was generally effective in triggering food-searching behaviour in both honey-fed (and thus conditioned to associate honey odour with food) and unfed, naive wasps. Mixing honey with imidacloprid and spinosad did not affect wasp responses. The remainin...

The Colorado potato beetle, Leptinotarsa decemlineata (Say), is one of the most damaging insect pests of potato, Solanum tuberosum L. Novaluron is a relatively new benzoylphenyl urea insect growth regulator with good activity against this pest. Earlier studies revealed that feeding on potato foliage treated with novaluron induces reversible egg hatch inhibition in adult Colorado potato beetles. We investigated whether novaluron effects depend on physiological state of the beetles at the time of exposure. The following four treatments were created: young beetles unmated at the beginning of the experiment and feeding on potato foliage treated with novaluron, young beetles unmated at the beginning of the experiment and feeding on untreated foliage, older beetles mated at the beginning of the experiment and feeding on foliage treated with novaluron, and older beetles mated at the beginning of the experiment and feeding on untreated foliage. The beetles were exposed to the respective treatments for 5 d. After that, both young and older beetles feeding on novaluron-treated leaves were switched onto untreated leaves and monitored for another 5 d to test their ability to recover. Young beetles unmated at the beginning of the experiment produced more eggs after feeding on the treated foliage, possibly indicating the presence of a pesticide-induced homeostatic modulation. No such effect was observed in the older beetles. Regardless of beetle physiological state at the beginning of the experiment, eggs produced on treated foliage did not hatch. The beetles eventually resumed laying viable eggs after being switched onto untreated foliage, with the recovery being delayed by approximately 24 h in young beetles compared with older beetles. Our results corroborate that novaluron reduces fertility of treated adults.

Interactions between lady beetles and the European fire ant (Myrmica rubra L.) tending potato aphids [Macrosiphum euphorbiae (Thomas)] were compared in the laboratory. Lady beetle species native to North America (Coccinella trifasciata perplexa Mulsant, Coleomegilla maculata lengi Timberlake, Hippodamia convergens Guérin-Méneville) and non-native species of Palearctic origin [Coccinella septempunctata L., Harmonia axyridis (Pallas), Hippodamia variegata (Goeze), Propylea quatuordecimpunctata L.)] were evaluated. Harmonia axyridis consumed a significantly greater number of aphids compared with all other species but C. septempunctata. Ant stings affected H. variegata and C. septempunctata to a greater extent than other species. Ants showed a significantly greater amount of aggression toward H. convergens and H. variegata compared with P. quatuordecimpunctata. P. quatuordecimpunctata, C. trifasciata, and H. axyridis reacted significantly less to ants compared with H. variegata, H. convergens, C. maculata, and C. septempunctata. Differences in interactions with natural enemies may explain, in part, the successful establishment of some non-native coccinellids in new habitats.

Several studies suggest the possibility that non-native lady beetles may have replaced native lady beetles in some agricultural habitats. There is relatively little information, however, about lady beetle species composition outside of agricultural habitats. Evans (2004) suggested that native species have retreated to nonagricultural habitats in response to the arrival of non-native lady beetles (habitat compression hypothesis). To test this hypothesis, a survey of lady beetles was conducted in 2004 and 2005 in different habitats in Maine. From May to October, lady beetles were sampled in a variety of agricultural and nonagricultural habitats. In total, 3,487 and 2,903 lady beetles were collected in 2004 and 2005, respectively. Non-native lady beetles were found in a variety of habitats, including the habitats that would have likely served as a refuge for native species if the habitat compression hypothesis applied to the surveyed areas. Native species were found in a higher proportion in agricultural habitats compared with nonagricultural habitats and in very low numbers in all of the habitats surveyed. Hippodamia tredecimpunctata tibialis (Say) and Coccinella transversoguttata Brown, the two native species that were once dominant here, made up only 1.09 and 0.07% of the total lady beetles collected, respectively. In this survey, we failed to detect evidence that native lady beetles have retreated to nonagricultural habitats in response to the arrival of non-native lady beetles.

The Colorado potato beetle, Leptinotarsa decemlineata (Say), is widely regarded as the most important insect defoliator of potatoes. Its current range covers about 16 million km2 in North America, Europe, and Asia and continues to expand. This insect has a complicated and diverse life history, which is well-suited to agricultural environments, and makes it a complex and challenging pest to control. Dispersal, closely connected with diapause, feeding, and reproduction, allow the Colorado potato beetle to employ “bet-hedging” reproductive strategies, distributing its offspring in both space (within and between fields) and time (within and between years). The Colorado potato beetle played a large role in creating the modern pesticide industry, with hundreds of chemicals tested against it. High selection pressure, together with natural propensity to adapt to toxic substances, eventually resulted in a large number of insecticide-resistant Colorado potato beetle populations. Since the middle of the last century, the beetle has developed resistance to 52 different compounds belonging to all major insecticide classes. Resistance levels vary greatly among different populations and between beetle life stages, but in some cases can be very high (up to 2,000-fold). Known mechanisms of Colorado potato beetle resistance to insecticides include enhanced metabolism involving esterases, carboxylesterases and monooxygenases, and target site insensitivity, as well as reduced insecticide penetration and increased excretion. There is also some evidence of behavioral resistance. Resistance mechanisms are sometimes highly diverse even within a relatively narrow geographical area. Resistance is usually inherited as an incompletely dominant or incompletely recessive trait, with one or several genes involved in its determination. Because of pleiotropic effects of resistant alleles, insecticide-resistant beetles often have reduced relative fitness in the absence of insecticides. Rotating different classes of insecticides and reducing insecticidal pressure on pest populations by provision of temporal and spatial refuges from exposure to toxins have been proposed to delay evolution of resistance. However, insecticide resistance in this insect will likely remain a major challenge to the pest control practitioners. Still limited understanding of beetle biology, its flexible life history, and grower reluctance to adopt some of the resistance management techniques create impediments to successful resistance management. Overcoming these obstacles is not an easy task, but it will be crucial for sustainable potato production.

Repeated use of neonicotinoid insecticides has resulted in the first reported cases of Colorado potato beetle (Leptinotarsa decemlineata (Say)) resistance to imidacloprid. In the laboratory we determined susceptibility of the imidacloprid-resistant Colorado potato beetles from a population in Southern Maine to other insecticides currently registered for use on potato. This population was about 30-fold resistant to imidacloprid and could not be effectively controlled by its applications. Control mortality was significantly higher for the imidacloprid-resistant larvae than for the susceptible larvae, suggesting that fitness disadvantages may be associated with the resistance trait. Resistant larvae exhibited significantly less mortality than susceptible larvae when exposed to cyfluthrin, carbaryl, azinphosmethyl, and methamidophos. Their susceptibility to oxamyl was also somewhat reduced, although it did provide nearly 100% mortality at the highest concentration tested. Disulfoton was highly toxic to the resistant larvae. Oxamyl killed about 40% of the adults in greenhouse assays with potted potato plants, altered their feeding behavior (fewer adults up on plants), and reduced defoliation by more than 90%. Disulfoton was not lethal to adults, but significantly suppressed their feeding. In field trials with the resistant population, oxamyl and imidacloprid + spinosad provided the best beetle control. Novaluron had no detectable effect on beetle densities. There was little difference between the plots treated with imidacloprid or thiamethoxam and the untreated control. Our results suggest that insecticide rotation may be a valuable option for managing imidacloprid-resistant Colorado potato beetle populations. We also had a good consistency between the results of the Petri dish, greenhouse, and field experiments, indicating that screening under laboratory confinement may be useful when developing initial recommendations to potato growers in areas affected by resistance to neonicotinoids.

Potato aphid, Macrosiphum euphorbiae (Thomas), is a highly mobile aphid species that dominates aphid communities in Maine potato fields and may contribute to virus transmission between potato plants. We studied effects of simulated rain, wind, mechanical raking, fungicide application, reflective mulch, and predator [lady beetle, Harmonia axyridis (Pallast)] on the interplant movement of wingless adult potato aphids in greenhouse experimental arenas that imitated small segments of a potato field. The number of aphids dispersing from the central plant in the arena after a tested perturbation was recorded. Experiments were repeated with 3- to 4-wk-old plants with nonoverlapping canopies and with 4- to 5-wk old plants with canopies overlapping within rows. Aphids moved between potato plants even when canopies did not overlap and without any environmental perturbations. However, more aphids moved between larger plants with overlapping canopies. Rain significantly encouraged aphid movement between plants with nonoverlapping canopies. Wind, rain, and mechanical raking significantly encouraged aphid movement between plants with overlapping canopies. Regardless of canopy overlap, most aphids moved within the rows of potato plants. However, there was also considerable movement between the rows, even though the aphids had to walk over bare soil.

Potato aphid, Macrosiphum euphorbiae (Homoptera: Aphididae), is an abundant potato pest and vector of potato leaf-roll virus and potato virus Y in Maine and other potato growing areas. We investigated the circadian rhythmicity of its movement towards host plant odor. Effects of daily cycle (day or night) and illumination (light or dark) on the proportion of aphids colonizing potato leaflets were determined in a Petri plate arena and in a Y-tube olfactometer. In Petri dishes, both daily cycle and light had a highly significant effect on plant colonization. Increasing temperature reduced aphid colonization of the leaflets. In the olfactometer, light had a significant effect on the proportion of aphids walking towards the host plant. Interaction between time and light was also statistically significant, with the effect of illumination being smaller during the day than during the night. Our results suggest that circadian rhythm in host-finding behavior of the potato aphid is regulated by both exogenous and endogenous mechanisms.

Colorado potato beetle, Leptinotarsa decemlineata (Say), is the most important insect defoliator of potatoes and is infamous for its ability to develop insecticide resistance. Sperm precedence is an important consideration in developing resistance management plans. We determined the precedence of sperm from a postdiapause spring mating by using irradiated sterile males. We also investigated whether spring mating (with or without fertilization) affects reproduction of overwintered females. Precedence of sperm from spring matings versus overwintered sperm from fall matings was almost complete. Some fertilization from fall matings did take place in this experiment, but it exceeded 10% in only one of the 22 pairings. Females mated only the previous fall laid fewer eggs than spring-mated females, but spring mating did not seem to improve the hatch rate of deposited eggs. On average, mated females started laying 1.6 d sooner than fall-mated females, but this difference was not significant. Hatch rate did not change with time. Precedence of spring matings over stored sperm can help compensate for assortative summer mating among resistant individuals due to differences in developmental time between refuges and treated areas.