Modeling long-term, stage-structured dynamics of Tribolium castaneum at food facilities with and without two types of long-lasting insecticide netting
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,Insecticide Netting In this study, we focused on two types of long-lasting insecticide netting (LLIN) that have been found to be effective for managing various stored product insect pests. One is an LLIN consisting of a polyethylene netting (2 × 2 mm mesh, D-Terrence, Vestergaard, Inc., Lausanne, Switzerland) with 0.4% deltamethrin active ingredient (a.i.), while the second one is Carifend® net (40 deniers with mesh size 97 knots/cm2; BASF AG, Ludwigshafen, Germany) containing 0.34% α-cypermethrin (a.i.).,Foundational Model We used a standard Lefkovitch matrix model to project population growth for Tribolium castaneum, with four life stages (e.g., egg, larva, pupa, and adult;(Lefkovitch,1965). In equation (1), the Leftkovitch matrix L matrix (4 × 4) represents the life-stage structure of T. castaneum which has an egg, larvae, pupae, and an adult, where only the adults contribute to the fecundity, F. By multiplying L with the population vector ni(t), where t is time step (e.g., generation) and i is a life stage, we obtain the resultant vector ni(t + 1), which reveals the distribution of individuals across different life stages in the subsequent time period. In equation (1), P1 represents the probability of staying in the egg stage and G1 is the probability of moving from the egg to the larval stage, P2 is the probability of staying in the larval stage, G2 is probability of moving from the larval stage to pupal stage, P3 is the probability of staying in the pupal stage, G3 is probability of moving from the pupal stage to adult, while P4 is the probability of staying in the adult stage (Figure 1).,Model Parameterization and Scenarios We simulated population outcomes for up to 15 generations by using the life table data for T. castaneum using the R package popbio. Survivorship, fecundity, and transition information for each stage were derived from the literature (summarized in Table 1). The developmental duration of eggs, larvae, and pupae were 3.82 ± 0.005, 22.81 ± 0.67, and 6.24 ± 0.071 days (Kollros,1944). The average life duration of the adult used in this study was 221.16 days (Park et al., 1961). We used 94 offspring for fertility from the study Park et al.,(1965) and 99% rate of eclosion from pupae to adult. In order to explore the sensitivity of the base model to changes in mortality and fecundity, both of these parameters were systematically varied from near zero to their maximum value given in the base model (e.g., F = 94, P4 = 0.871). The parameters were varied alone or in combination and the resulting population growth was plotted. All plots were created using ggplot2 (Wickham, 2016) in R software (R Core Team, 2022). Three empirical scenarios from the literature were modeled containing estimates of fecundity reduction only, survivorship reduction only, or both fecundity and survivorship reduction when using LLIN (R.V. Wilkins et al., 2021; Gerken et al., 2021;Scheff et al., 2021, Scheff et al., 2023; Table 2). An individual projection matrix was constructed for each of the three scenarios and combinations of the reductions in fecundity, survivorship, or both. Population growth and proportion in each life stage was projected for 15 generations for each case, including the base model. Overall variation and oscillation were calculated to compare trends among proportion of life stages in each case. In order to compare differences in population sizes between cases for all generations and for generation 15 only, population sizes for each generation were bootstrapped 1000 times to provide iterative replication. The bootstrapped data were then compared one case to another using proc ttest in SAS (Version 9.4) for all generations and for generation 15 only. In addition, a sensitivity analysis was performed to determine which stage should be targeted to most greatly affect the population growth after exposure to the netting. Moreover, a mortality function based on empirical data with LLIN exposure collected in the laboratory
Data from: Direct lethality and time-delayed sublethal effects of multiple types of insecticide netting against stored product insects
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,Experimental Insects,The field strains of T. castaneum and R. dominica (F.) were used in this study. The former originates from Eastern Kansas in 2012, and the latter is also from Eastern Kansas but from 2019. For all species, four to eight-week-old adults were used. Rearings were kept at the USDA Center for Grain Animal Health Research in Manhattan, KS. Tribolium castaneum was reared on a mixture of 95% unbleached, organic flour and 5% brewer’s yeast, while R. dominica was reared on tempered organic whole wheat. Colonies were maintained at 27.5°C, 65% RH, and 14:10 for maintenance or 16:8 (L:D) h photoperiod for the experiment.,Treatments,Treatments included exposure to three different types of long-lasting insecticide-incorporated netting (LLIN). These consisted of 1) Carifend®, LLIN with 0.34% alpha-cypermethrin (40 deniers, BASF, Ludwigshafen, Germany), 2) D-Terrence, LLIN with 0.4% deltamethrin (2 × 2 mm mesh, Vestergaard SA., Lausanne, Switzerland), and 3) 8% etofenprox LLIN (AgBio, Inc, CO, USA), and for control, we used netting identical to the Carifend or Vestergaard netting but lacking insecticide.,Direct Lethality Assessments,Cohort of 20 mixed-sex adult beetles were exposed for 5, 60, or 120-min intervals on netting affixed to a 9 × 9 cm2 petri dish in the laboratory. After exposure, we took the evaluated condition after 0, 24, 72, or 168 h as alive, affected, or dead condition (Figure 1), according to the definitions described in Ranabhat et al. (2022) in Petri dishes without netting containing 8.5 cm D filter paper. Briefly, living adults were defined as moving with normal speed and activity and able to right themselves if flipped. By contrast, affected adults exhibited sluggish or drunken movements, could not right themselves if flipped, and some or all of their limbs exhibited twitching. Dead adults were completely immobile. For post-exposure treatment, adults were held under the same environmental chamber conditions as the colonies but without supplemental food after exposure. We performed a total of n = 4 replications per treatment combination for each species.,Baseline Mobility Assay after Exposure to LLINs,Based on the observation of the lethality assay, we focused our baseline mobility assay on Carifend® and D-Terrence LLIN. Using only alive adults, we assessed their movement in six individual Petri dishes (100 × 15 mm D: H) that consisted of a filter paper (85 mm D, Grade 1, GE Healthcare, Buckinghamshire, United Kingdom) lining. Treatments included a negative control (e.g., filter paper only), one of the two LLINS, or an identical netting to the Carifend or Vestergaard netting but without insecticide (e.g., as a positive control). Their movement was tracked for 60-min using a network camera (GigE, Basler AG, Ehrenburg, Germany) affixed 80 cm above the dishes. The Petri dishes were backlit using a LED light box (42 × 30 cm W: L, LPB3, Litup, Shenzhen, China) to increase contrast and affixed in place with white foam board. The video was streamed to a computer and processed in Ethovision (v.14.0, Noldus Inc., Leesburg, VA). The program automatically calculated the total distance moved (cm) and the instantaneous velocity (cm/s) over the 60-min period for each adult. Each adult was considered a replicate and was never used more than once. In total, n = 18 replicates were performed per treatment combination.,Comparison of Sublethal Effects among LLINs,For the sublethal movement assay, mixed-sex adult beetles were exposed to the Carifend®, D-Terrence LLIN, or control net as mentioned above. Cohorts of 5–10 adults were exposed for 5- or 60-min intervals on LLINs affixed to a 9 × 9 cm2 Petri dish in the laboratory. After exposure, the effects of the LLINs on adult movement were assessed either immediately or after 72 h in Petri dishes under the same environmental chamber conditions as the colonies but without supplemental food and then assayed using the video-tracking system described above by using Ethovision
Data from: Direct lethality and time-delayed sublethal effects of multiple types of insecticide netting against stored product insects
공공데이터포털
,Experimental Insects,The field strains of T. castaneum and R. dominica (F.) were used in this study. The former originates from Eastern Kansas in 2012, and the latter is also from Eastern Kansas but from 2019. For all species, four to eight-week-old adults were used. Rearings were kept at the USDA Center for Grain Animal Health Research in Manhattan, KS. Tribolium castaneum was reared on a mixture of 95% unbleached, organic flour and 5% brewer’s yeast, while R. dominica was reared on tempered organic whole wheat. Colonies were maintained at 27.5°C, 65% RH, and 14:10 or 16:8 (L:D) h photoperiod.,Treatments,Treatments included exposure to three different types of long-lasting insecticide-incorporated netting (LLIN). These consisted of 1) Carifend®, LLIN with 0.34% alpha-cypermethrin (40 deniers, BASF, Ludwigshafen, Germany), 2) D-Terrence, LLIN with 0.4% deltamethrin (2 × 2 mm mesh, Vestergaard SA., Lausanne, Switzerland), and 3) 8% etofenprox LLIN (AgBio, Inc, CO, USA), and for control, we used netting identical to the Carifend or Vestergaard netting but lacking insecticide.,Direct Lethality Assessments,Cohort of 20 mixed-sex adult beetles were exposed for 5, 60, or 120-min intervals on netting affixed to a 9 × 9 cm2 petri dish in the laboratory. After exposure, we took the evaluated condition after 0, 24, 72, or 168 h (as alive, affected, or dead condition, according to the definitions described in Ranabhat et al. (2022) in Petri dishes without netting containing 8.5 cm D filter paper. Briefly, living adults were defined as moving with normal speed and activity and able to right themselves if flipped. By contrast, affected adults exhibited sluggish or drunken movements, could not right themselves if flipped, and some or all of their limbs exhibited twitching. Dead adults were completely immobile. For post-exposure treatment, adults were held under the same environmental chamber conditions as the colonies but without supplemental food after exposure. We performed a total of n = 4 replications per treatment combination for each species.,Baseline Mobility Assay after Exposure to LLINs,Based on the observation of the lethality assay, we focused our baseline mobility assay on Carifend® and D-Terrence LLIN. Using only alive adults, we assessed their movement in six individual Petri dishes (100 × 15 mm D: H) that consisted of a filter paper (85 mm D, Grade 1, GE Healthcare, Buckinghamshire, United Kingdom) lining. Treatments included a negative control (e.g., filter paper only), one of the two LLINS, or an identical netting to the Carifend or Vestergaard netting but without insecticide (e.g., as a positive control). Their movement was tracked for 60-min using a network camera (GigE, Basler AG, Ehrenburg, Germany) affixed 80 cm above the dishes. The Petri dishes were backlit using a LED light box (42 × 30 cm W: L, LPB3, Litup, Shenzhen, China) to increase contrast and affixed in place with white foam board. The video was streamed to a computer and processed in Ethovision (v.14.0, Noldus Inc., Leesburg, VA). The program automatically calculated the total distance moved (cm) and the instantaneous velocity (cm/s) over the 60-min period for each adult. Each adult was considered a replicate and was never used more than once. In total, n = 18 replicates were performed per treatment combination.,Comparison of Sublethal Effects among LLINs,For the sublethal movement assay, mixed-sex adult beetles were exposed to the Carifend®, D-Terrence LLIN, or control net as mentioned above. Cohorts of 5–10 adults were exposed for 5- or 60-min intervals on LLINs affixed to a 9 × 9 cm2 Petri dish in the laboratory. After exposure, the effects of the LLINs on adult movement were assessed either immediately or after 72 h in Petri dishes under the same environmental chamber conditions as the colonies but without supplemental food and then assayed using the video-tracking system described above by using Ethovision software. Only adults classified as alive or
Data from: Effects of synergists on the efficacy of long-lasting insecticide-incorporated netting against Tribolium castaneum (Coleoptera: Tenebrionidae) and Rhyzopertha dominica (Coleoptera: Bostrichidae)
공공데이터포털
,Experimental Insects,Adult Tribolium castaneum and Rhyzopertha castaneum were obtained from insect colonies kept at the United States Department of Agriculture (USDA) Center for Grain and Animal Health Research in Manhattan, Kansas. Tribolium castaneum was continuously reared on 95% unbleached, organic wheat flour with 5% brewer’s yeast added, while R. dominica was reared on organic wheat, and both were held in an environmental chamber (Percival Scientific, Model CTH-811, Perry, IA, USA) set at a temperature of 30 °C, relative humidity (RH) of 65%, and a 16/8 h light/dark photoperiod. For the bioassays below, 3- to 4-week-old T. castaneum and R. dominica adults were used.,Synergist-coated Vials,Insecticide synergists tested in the bioassays were: piperonyl butoxide (PBO, Tokyo Chemical Industry Co. Ltd., Tokyo, Japan), diethyl maleate (DEM, Thermo Scientific Chemicals, Waltham, MA, USA), and triphenyl phosphate (TPP, Sigma-Aldrich, St. Louis, MO, USA). PBO, DEM, and TPP were dissolved in acetone, separately, and the concentrations used for them were all 0.1 mg/ml. Glass scintillation vials (27 × 61 mm, D × H, Wheaton Science Products, Millville, NJ, USA) were coated with 0.5 ml of synergist solution by rolling the vials on a Roto-Torque Heavy Duty Rotator (Model 7637, Cole-Parmer Instrument Company, Vernon Hills, IL, USA) to spread the synergist across the inner surface until all visible signs of liquid had disappeared. The vials were then left open to evaporate the acetone residues. In parallel, vials were coated with 0.5 ml of acetone as a control.,Long-lasting Insecticide-incorporated Netting (LLIN),Carifend® LLIN incorporating 0.34% (w/w) alpha-cypermethrin (BASF, Ludwigshafen, Germany) was used in this study. This netting was cut into squares and secured to the bottoms of square Petri dishes (100 × 100 × 15 mm, L × W × H, VWR, Radnor, PA, USA) as LLIN exposure arenas.,Effects of Synergists on LLIN against T. castaneum and R. dominica,Synergists and LLIN exposure bioassays for T. castaneum and R. dominica adults were conducted as below. Twenty adults were pre-exposed in a glass scintillation vial coated with one of the three synergists (PBO, DEM, TPP, or acetone as a control) for 1 h, and then transferred to a LLIN exposure arena. For T. castaneum, twenty adults were exposed to LLIN for 6, 12, 24, 48, 72, 96, 120, 144, and 168 h, respectively, and then examined for adult condition (unaffected, affected, and dead). For R. dominica, preliminary experiments showed that adults were very susceptible to LLIN, so twenty adults were exposed to LLIN for 0.5, 1, 2, 6, 12, 24, and 48 h, respectively. The adults were then transferred to a 35 × 10 mm Petri dish (Falcon, Franklin Lakes, NJ, USA) and adult condition was examined after 24 h. T. castaneum and R. dominica adults were observed and recorded as unaffected if they were active and behaving normally with coordinated walking and species-specific movements; affected if they had uncoordinated waliking and sluggish movements or were on their backs with legs twitching, and/or could not right themselves after being prodded (i.e., knocked down); finally, recorded as dead if they were completely motionless even after prodding. There were 4 replicates for each treatment combination of synergist type × LLIN exposure time.,Effects of Food on Efficacy of Synergists and LLIN Combined Exposures in T. castaneum and R. dominica,Twenty adults were pre-exposed in a vial coated with PBO, DEM, TPP, or acetone as a control for 1 h, and then exposed to LLIN for 72 h (T. castaneum) and 1 h (R. dominica). After exposure, the adults were transferred from LLIN exposure arena to a recovery arena, consisting of Petri dish (35 × 10 mm, D × H, Falcon, Franklin Lakes, NJ, USA) with 0.5 g of wheat flour or without flour as a control. Adult condition was assessed as number of unaffected, affected and dead at 1, 3, 5, and 7 d post-exposure. The delayed mortality and recovery rates were expressed as percentages,
Data from: Efficacy of long-lasting insecticide-incorporated netting in controlling preharvest and postharvest pest insects: a meta-analysis study
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,This dataset aims to provide a comprehensive analysis of existing literature on the utilization of LLINs in the management of pre- and postharvest pest insect species (excluding those in urban systems and vectors). By synthesizing findings from a broad spectrum of studies, we aim to discern patterns relative to system and experimental design and develop a new synthesized understanding of the effectiveness of LLINs in agriculture. Moreover, our dataset intends to identify and address gaps in current knowledge. Through a meta-analytical approach, this review aims to distill key insights that can inform future research directions, and practical applications in the integrated management programs of pre- and postharvest insect pests.,,The goal of this dataset is to understand the efficacy of long-lasting insecticide netting in agriculture. We used databases such as Web of Science, Scopus, and Google Scholar to find studies related to LLIN. First, we carried out a literature search on the topic of insecticide netting. We used various Keyword combinations to identify a broad initial dataset, including various terms such as ‘netting’, ‘LLIN’ ‘insecticide netting’ in combination with either ‘insects’, ‘preharvest’, ‘postharvest’, and ‘agriculture’. Papers were individually checked to ensure they met criteria for inclusion in the meta-analysis, which included that 1) articles were published between 1990–2024, 2) the focus was on agricultural crops (not urban systems or vectored diseases), 3) netting was not solely used for exclusion (e.g., in the absence of an insecticidal ingredient), and 4) included a proper negative or stakeholder standard control. At the end, this left us with a total of 44 peer-reviewed publications (23 postharvest + 22 preharvest studies, one common study for pre and postharvest insects) with 285 data points on the efficacy of LLINs in agriculture. However, an additional required criterion was that all studies needed to present means, standard errors, and samples sizes for both controls and LLIN treatment. After this additional criterion, there was a final total of 31 peer-reviewed publications (19 postharvest + 12 preharvest studies). Additionally, potential publication bias was assessed by using funnel plots, and statistical tests were evaluated.,
Data from: Laboratory evaluation of two insecticide-incorporated nets for the management of Prostephanus truncatus
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,Insects,Adult P. truncatus were obtained from insect colonies kept in the Laboratory of Entomology and Agricultural Zoology (LEAZ), at the Department of Agriculture, Crop Production and Rural Environment, University of Thessaly, Greece, on whole maize kernels, at 26°C and 55% relative humidity (RH) and continuous darkness.,Insecticide-Incorporated Netting,The experiments were carried out in plastic petri dishes 90 mm in diameter (50.4 cm2 bottom surface). The inside of each petri dish was covered with two types of LLIN (0.4% deltamethrin, D-Terrence, Vestergaard INC., Lausanne Switzerland; and 0.34% alpha-cypermethrin, Carifend, BASF Ag, Ludwigshafen, Germany) and a polytetraflurorethylene preparation (Fluon, 60 wt% dispersion in water, Sigma-Aldrich Chemie GmbH, Steinheim, Germany) to prevent insects from escaping. An additional series of dishes with physically identical control netting were prepared without an insecticide treatment to serve as the control. Twenty P. truncatus were then exposed on the insecticide-treated netting in petri dishes for 60, 90, 120, 240 min, 1, 3, and 5 days.,Mortality and Recovery,After exposure, insects were evaluated for mortality and individual P. truncatus that remained alive or knocked down (not dead) were then place into clean Petri dish arenas with a small amount of clean cracked maize kernels and evaluated for delayed mortality after 7 days. Using a stereomicroscope (SMZ-18, Nikon Inc., Tokyo, Japan) under 60× magnification, P. truncatus were classified as alive (moving normally, is able to right itself when flipped over, no twitching), affected (moving sluggishly or erratically, unable to right itself, twitching of antennae or legs may be present), or dead (completely immobile even after prodding) according to prior published definitions. There were three 3 replicates for each time treatment and netting type and 3 subreplicates for a total of 9 replicates for each treatment combination.,Explanation of files,The file "Greece Net Data 2023_All" includes the raw mortality ratings, whereas "Greece Net Data 2023_Recovery" includes calculated recovery values by P. truncatus, where recovery was calculated to "alive" from the initial reading (0 d) in "Greece Net Data 2023_All".,
Data from: Pairing residual contact insecticide use with long-lasting insecticide-incorporated netting to reduce dispersal and damage by stored product insects
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,2.1 Experimental Insects,A field strain of the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) and the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) obtained from Pottawatomie Co, KS, and eastern KS, respectively, were used in this study. We used four to eight-week-old adults of both species. Cultures of R. dominica and T. castaneum have been maintained in the laboratory since 2019 and 2012, respectively at the USDA Center for Grain Animal Health Research in Manhattan, KS. Ryzopertha dominica were reared on tempered organic whole wheat, while T. castaneum were reared on a mixture of 95% unbleached, organic flour and 5% brewer’s yeast. The colonies were subcultured on a monthly basis. The colonies were maintained at 27.5°C, 65% RH, and 14:10 (L:D) h photoperiod in environmental chambers (Percival Scientific, Perry, IA, USA).,2.2 Experimental Arena and its Production,The experimental arena consisted of a 63 cm × 15.5 cm × 9.5 cm L:W: H metal frame. This arena also contained four square blocks of concrete (e.g., Rockite, Hartline Products Co., Cleveland, OH, USA) measuring 15.24 cm × 15.24 cm × 1.5 cm L:W: H to create a testing platform that mimics the surface of a food facility (Figure 1). The concrete was prepared by first mixing tap water and Rockite cement mix in a large water pitcher. The tap water was added to the dry Rockite mixture and combined until a thick paste consistency was achieved. The slurry was poured into a 1.1 L volume silicone square mold (15.24 cm × 15.24 cm). The slurry was poured 1.5 cm thick. The cement concrete squares were left to dry and solidify at room temperature for 2–3 d. Cement squares were modular and new ones were used with each replicate performed for the assay.,In each experimental arena, there were four cement squares (Figure 1). We applied tape (VWR International, LLC Radnor, PA, USA) on the wall of the metal frame before placing the concrete to make it easier to remove the concrete after each replicate was conducted, and so the metal forms could be re-used. Adhesive caulk (DAP Kwik Seal, DAP Products Inc., Baltimore, MD, USA) was applied to fill any gaps between the concrete squares or between the concrete and the metal frame. The concrete at the distal end of the experimental arena was filled with 20 g of whole, organic, unbleached flour (Heartland Mills, Marienthal, KS, USA) for T. castaneum or organic whole wheat (Heartland Mills, Marienthal, KS, USA) for R. dominica. Insects were released on the concrete square at the opposite end. The two middle concrete slabs were reserved for one or two treatments as below. The inner wall of the metal frame was coated with fluon (polytetrafluoroethylene, Sigma-Aldrich Co., St. Louis, MO, USA) to prevent insect escape.,2.3 Insecticide formulation for concrete,2.4 Treatment,We assembled the experimental arenas for each treatment replicated on separate days. In each experimental arena, we released 30 mixed-sex adults of either R. dominica or T. castaneum (just a single species per arena) at the far end of the testing platform opposite the food source. Each dispersal apparatus was placed into a sterilite bin (86.3 × 30.5 × 39.4 cm L:H:W) to prevent insect escape and effects from neighboring apparatuses, and placed on the shelf of a walk-in environmental chamber at the same conditions as the rearing colonies. After a 48 h dispersal period, we counted the number of insects on each concrete square and we also checked their conditions in clean petri dishes lined with filter paper. The condition of the adults was checked under the dissecting microscope and rated as alive, affected, or dead by following the definitions in Morrison et al. 2018. Briefly, the alive were moving around normally without impairment, while those that were affected showed uncertain movements, twitching of extremities, and/or were not able to right themselves after being gently prodded with a paintbrush. Those that were classified
Data from: Spillage and food dust do not decrease efficacy of long-lasting insecticide-incorporated netting against stored product insects
공공데이터포털
,Insect Sources,Insect colonies of R. dominica and T. castaneum maintained continuously at the USDA-ARS Center for Grain and Animal Health Research were used. This included T. castaneum collected in Eastern KS (USA) from 2012, and R. dominica collected from Eastern KS in 2019. Tribolium castaneum and had been reared on a mixture of 95% unbleached, organic flour and 5% brewer’s yeast, while R. dominica was reared on tempered organic whole wheat. Adults that were 4–6-week-old were used for experiments. Colonies were maintained at 27.5°C, 65% RH, and 14:10 (L:D) h photoperiod.,Treatments,The following netting treatments were used: negative control (e.g., no netting), positive control (netting identical to LLIN but without insecticide; Item#1721-9668, Casa Mesh White, Casa Solid, Joann’s Fabrics, Hudson, OH, USA), 0.34% w/w alpha-cypermethrin LLIN (Carifend, BASF Corps, Ludwigshafen, Germany), and a 0.4% w/w deltamethrin LLIN (D-Terrence, Vestergaard Inc., Lausanne, Switzerland).,Laboratory food dust assay,To evaluate the effect of food dust on the efficacy of LLIN, there were two food dust regimes. Netting was either used as is or fully dipped into organic flour (Heartland Mills, Marienthal, KS, USA) that filled a 9 × 9 cm square Petri dish. After exposure to food dust, the netting was used to line a new, clean 9 × 9 cm Petri dish. Rhyzopertha dominica and T. castaneum adults were tested in cohorts of 20 and exposed on the netting for 10 min continuously in the Petri dishes, then their conditions were checked at 1, 24, 48 h, and 168 h after exposure. Insects were held in an environmental chamber set to 27.5°C, 65% RH, and 14:10 L:D. Conditions were classified as the percentage that were alive (normally moving around unimpeded), affected (showing abnormal or sluggish movements, but movement still present, even if just twitching of extremities), or dead (completely immobile; full definitions in Morrison et al. 2018). This was performed under a stereomicroscope (SMZ18, Nikon Inc., Tokyo, Japan). A total of n = 5 replicate cohorts were tested per combination of treatments (dust regime, netting type, exposure time, post-exposure holding duration, and species).,Spillage assay,To evaluate whether netting could be used to protect sites of spillage, we performed a spillage assay in the laboratory. For this assay, only netting without insecticide but identical to LLIN and 0.34% alpha-cypermethrin LLIN (BASF) was used. Netting was placed covering a single layer of 35 g of whole organic hard winter wheat (Heartland Mills, Marienthal, KS, USA) in a 9 × 9 cm square Petri dish. A control treatment included a single layer of positive control or alpha-cypermethrin LLIN placed in a Petri dish without food. Cohorts of 20 mixed-sex R. dominica or T. castaneum adults were exposed continuously to the netting for 48 h. After that period, the conditions of the adults were recorded as alive (moving normally), affected (sluggish movements, unable to right themselves when fallen, or twitching body parts), or dead (completely immobile) according to established definitions in (Morrison et al., 2018). After sieving adults, we placed the grain from the Petri dish in a separate vial (11 × 4.9 cm H:D) for six weeks to check for progeny production, including the number of larvae, pupae, and adults. A total of n = 7 replicate cohorts were tested per combination of treatments.,Interception assay,To determine whether LLIN can prevent horizontal dispersal of stored product insects to sites of spillage, we performed an interception assay. A single layer of organic, whole wheat (Heartland Mills, Marienthal, KS, USA) was placed in a 245 × 245 mm large square Petri dish (Item# 431111, Corning Inc., Corning, NY, USA). In the center of the dish, a 2.5 x 24.5 cm (W × L) strip of netting was added on top of the wheat. A total of 50 mixed-sex R. dominica or T. castaneum adults were added to the middle of zone 1 (e.g., release zone; Figure 1). The remainder of the dish was