,Four to six-week-old larvae of Trogoderma variabile and Trogoderma inclusum were used for the experiment. Both strains were originally obtained from the field in north-central Kansas in 2016 and 2012, respectively. Colonies of these species were reared under controlled conditions in an environmental chamber set to a temperature of 27.5 °C, 65% RH, and 14:10 (L:D) h photoperiod. Both species were fed 300 g of ground dog food (SmartBlend, Lamb flavor, PurinaOne, St. Louis, MO, USA) with oats sprinkled on top and a moistened, crumpled paper towel placed on the surface in a 950-ml mason jar.,Treatments The long-lasting insecticide-incorporated polyethylene netting (2 × 2 mm mesh, D-Terrance, Vestergaard Inc., Lausanne, Switzerland) included 0.4% deltamethrin, or control netting that was identical in physical properties but without insecticide. These were used with the movement assay. We assessed the movement in the vicinity of important pheromonal and food kairomones after exposure to LLIN or control netting. Food consisted of 0.01 g of organic, unbleached flour (Heartland Mills, Marienthal, KS, USA), and pheromonal stimuli included a broad spectrum, multi-species lure (PTL lure, IL-108-10, Batch#1288200321, Insects Limited, Westfield, IN, USA), including Trogoderma spp pheromone (Ranabhat et al. 2023a). In each replicate, we used a single pellet (white color), and affixed it in place so it did not move in a Petri dish using a 1 × 1 mm square of parafilm. For each replication of testing, we used a fresh lure.,Movement Assay The movement of larvae after exposure to the 0.4 % deltamethrin LLIN or a control netting in response to food cues (using 0.01 g of flour) or with conspecific sex pheromones (using a single bead from a disaggregated PTL lure held in place with a small square of parafilm), was tracked in six individual arenas (100 × 15 mm D: H) with a piece of filter paper (85 mm D, Ahlstrom-Munksjö, Helsinki, Finland) lining the bottom for 30 min using a network camera (GigE, Basler AG, Ahrenburg, Germany) affixed 76 cm above and centered over 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.14.5 Noldus Inc., Leesburg, VA, USA). Prior to use in the movement assay, larvae of T. variabile or T. inclusum were exposed to the 0.4% deltamethrin LLIN or a control netting for 1 min in a 21 × 21 cm square Petri dish, then their movement was tracked individually after a post-exposure holding duration of 1 min or 24 h. A small 1.1 cm hidden stimulus zone encircled each stimulus, midway and centered on each half of the arena wherein movement was tracked separately from each half of the arena (control vs. treatment). The total distance moved (cm), instantaneous velocity (cm/s), frequency of entering each half of the petri dish and stimulus zone, cumulative duration spent in each zone (s), and latency of entering each zone (s) over a 30 min trial period was logged after exposure to a given treatment. The control side of the arena remained empty. A total of n = 16 replicates were run per treatment combination for both species,No-Choice Release-Recapture Assay A release- recapture experiment was conducted for the larvae of both T. variabile and T. inclusum where larvae were exposed to the 0.4% deltamethrin LLIN and control netting for 1 min. After exposure, treated insects were released at one corner of the sanded plastic bin (60 × 41.6 × 16.5 cm L:W:H ). A commercial pitfall trap (Dome Trap™, Trécé, Inc., Adair, OK, USA) that contained a PTL lure (used only white beads as above), or 0.01 g flour, or no stimuli (unbaited for control), was deployed in the opposite corner, diagonally across from the release point in the bin. The bins were located in a large (4.8 × 2.1 × 6 m, L:W:H) walk-in environmental chamber (Percival Instruments, Dallas County, IA,

,The aim of the current study was to track the movement of phosphine-resistant and -susceptible adults of the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), which is a major pest of stored products, after brief exposures to phosphine. Exposures were followed for extended intervals to assess the recovery patterns, and how those patterns are related to known resistance to phosphine. A video-tracking procedure coupled with Ethovision software was used to assess movement after exposure.,Two strains of T. castaneum were used, one susceptible and one resistant to phosphine. The susceptible T. castaneum strain had been maintained in continuous culture without any known exposure to phosphine for >30 years at the USDA-ARS Center for Grain and Animal Health Research (CGAHR), in Manhattan, KS, USA. The phosphine-resistant strain of T. castaneum was collected from wheat in Palmital, Brazil during 1988 (BRZ-5). The rearing media consisted of 95% organic, unbleached, wheat flour plus 5% brewer's yeast. Tribolium castaneum were reared under laboratory conditions of 27.5°C, and 65% relative humidity (R.H.), 14:10 L:D. Adults, of mixed sex and <1 month old, were used in the exposure bioassays.,The protocol that was used in our bioassays to generate phosphine was the Phosphine Tolerance Test (Detia Degesch GmbH, Laudenbach, Germany) with some modifications, as performed by Agrafioti et al. 2021. In particular, the phosphine was generated within a plastic canister (5 L capacity) by adding 50 mL of water to two kit magnesium phosphide pellets. The concentration of phosphine gas inside the plastic canister was determined by using several dosimeter Draeger glass tubes (Draeger 25A, 0–10 000 ppm, Draeger Safety AG & Co., USA). Ten adults of each strain were placed in a plastic syringe of 100 mL with separate syringes used for each species and strain. Then, a specific gas quantity was removed from the canister with the syringe and blended with fresh air to produce a 100-mL volume with a concentration of either 1000 or 3000 ppm and compared to phosphine-free controls (0 ppm). The insects inside the syringe were held at the concentrations above for a 5 min exposure, while additional syringes containing only fresh air and insects were used as negative controls.,To understand the propensity for movement after a 5 min phosphine exposure, a video-tracking procedure was used. After exposure of phosphine-resistant or phosphine-susceptible T. castaneum for 5 min, adult movement was evaluated immediately after exposure or 24 h later under the same environmental chamber conditions as the colonies (see Source Insects), but held without supplemental food. Movement was recorded for 3 h immediately after phosphine exposure but binned into 30 min intervals (e.g., 0–30, 30–60, 60–120, 120–150, and 150–180 min) in order to evaluate how movement varied over the measured time period. Movement was also recorded 24 h after exposure for periods of 1 h (binned by 30 min intervals). Movement measures of adults was tracked in six replicate Petri dishes (90 × 15 mm D:H) with a piece of filter paper (85 mm D, Grade 1, GE Healthcare, Buckinghamshire, United Kingdom) lining the bottom using a network camera (GigE, Basler AG, Ahrenburg, 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 with holes specifically cut to size for the petri dishes. Video was streamed to a nearby computer and processed in Ethovision (v. 14.0.1322, Noldus Inc., Leesburg, VA). The software was used to calculate the total distance moved (cm) and the mean instantaneous velocity (cm/s) for each adult. Each adult was considered a replicate and was never used more than once. Only adults classified as alive (normal movement speed and activity), or affected (sluggish movements or on back with legs twitching) were used in this assay.

,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

,We compared all combinations of three commercial traps and five different attractants on the capture of stored-product insects for two consecutive years in three food processing facilities in Central Greece. Specifically, Facility 1 and 2 were pasta factories and Facility 3 was a flour mill. The traps that were used in the experiments were Dome Trap (Trécé Inc., USA), Wall Trap (Trécé Inc., USA) and Box Trap (Insects Limited, Ltd., USA). The attractants that were evaluated were 0.13 g of : 1) of PantryPatrol gel (Insects Limited, Inc., USA), 2) Storgard kairomone food attractant oil (Trece Inc.), 3) wheat germ (Honeyville, USA), 4) Dermestid tablet attractant (Insects Limited Inc., USA). The traps were inspected approximately every 15 days and rotated clockwise. The captured insects were transferred to the Laboratory of Entomology and Agricultural Zoology (LEAZ) at University of Thessaly for identification. The results indicated that there was a wide range of species within the three facilities throughout the trapping period, with the Indian meal moth, Plodia interpunctella (Hübner), the red flour beetle, Tribolium castaneum (Herbst) and the cigarette beetle, Lasioderma serricorne (F.), being the most abundant. Although there were noticeable differences among the different traps and attractants for the capture of certain species, all combinations provided comparable population fluctuation patterns. In general, Dome traps, baited with either the oil or the gel, were found to be the most effective.,There are not much data available so far for the simultaneous comparable use of different trapping devices and different attractants in commercial facilities for long-term monitoring. Certain lures are marketed toward particular pests or classes of pests, while others might be more generic, multi-species lures. To shed light on this issue, we evaluated a series of combinations of floor traps and attractants, in three commercial facilities in Greece, for a period of two years. Our questions included both which trap was broadly most effective as well as whether different combinations of traps and types of attractants were delivering novel information about the stored product insect community. The traps include two types of floor traps, and the wall trap used in the USDA khapra beetle detection programs. The lures included the Insects Limited ™ dermestid tab that is more specifically focused on food kairomones for only that taxon, and the same company’s PantryPatrol gel, which uses wheat kairomones and the pheromones of multiple species, including dermestids. We also use the Trécé Storgard kairomone oil, and simple wheat germ, which are both multi-species kairomones with no pheromones.,Resources in this dataset:,Resource Title: 2018 and 2019 field trapping data,File Name: kb_greek_data_ag_data_commons.csv,Resource Description:,2.1 Storage facilities: The storage facilities in which this study took place are located in Central Greece. The selection of these facilities was based on their size, the accessibility from University of Thessaly (UTH) personnel and the known historical presence of stored product insect species and other arthropods. The sampling was conducted in three types of storage facilities refereed as Facility 1, Facility 2 and Facility 3. Facilities 1 and 2 are pasta factories, with substantial quantities of soft and hard wheat, flour and bran, but also some barley and maize, while Facility 3 is a flour mill, mostly focused on soft wheat processing. The deployment of the traps on each facility was conducted at 18 June 2018, 4 July 2018, and 3 July 2018 for Facility 1, 2 and 3, respectively.,2.2. Traps, attractants and inspection: The trap types that were used in our experiments were Dome Trap (Trécé Inc., USA), Wall Trap (Trécé Inc., USA) and Box Trap (Insects Limited, Ltd., USA). These traps have been proven effective for monitoring purposes based on previous studies (Toews et al., 2009; Athanassiou and Arthur, 2018;

,Reproductive output and behavioral data from stored-product pest insects exposed to long-lasting insecticide treated netting. Includes offspring output and survival for Trogoderma variabile (treated as larvae) and Tribolium castaneum (treated as adults).,,

,Attraction Assessment - assessment of different lure sources, including pheromones and kairomones for red flour beetle and lesser grain borer in the wind tunnel and release-recapture experiment under controlled settings. Abbreviations: WGO - wheat germ oil; Tab - Insects Limited SPB tablet bait; NC - negative control (no stimulus); DDGS - dried distiller's grains with soluables. There were a total of n = 12 replicate releases for the release-recapture and n = 30 replicate individuals for the wind tunnel per treatment.,Dose Dependency Data - evaluation of whether dose-dependency in attraction exists for red flour beetle and lesser grain borer in the wind tunnel and in a release-recapture experiment for the Insects Limited SPB lure. Abbreviations: SPB1 - a single Insects Limited SPB tablet bait; SPB2 - two Insects Limited SPB tablet baits; SPB3 - three Insects Limited SPB tablet baits; Ctrl - negative control (no stimulus); DDGS - dried distiller's grains with soluables. There were a total of n = 24 replicate releases for the release-recapture and n = 30 replicate individuals for the wind tunnel per treatment.,Spillage Trap Collections - datasheet for the number and lowest taxonomic unit of insects collected in interception traps with different kill mechanisms and stimuli at three food facilities in Arkansas and Kansas during 2018 and 2019. There were a total of 27 taxa tracked, and captures totaled to near 4,000. Abbreviations: C, control netting only (no stimulus); L, insecticide-netting only (no stimulus); LS, insecticide-netting with a single SPB Insects Limited tab lure; CS, control netting with a single SPB Insects Limited tab lure. State abbreviations: AR - Arkansas, KS - Kansas. There were three transects per site, each with every treatment above represented, thus a total of n = 8-9 replicate deployments in AR and n = 12 deployments in KS.,Spillage Trap Progeny- datasheet for progeny production in interception traps after six weeks under constant conditions with different kill mechanisms and stimuli at three food facilities in Arkansas and Kansas during 2018 and 2019. Abbreviations: C, control netting only (no stimulus); L, insecticide-netting only (no stimulus); LS, insecticide-netting with a single SPB Insects Limited tab lure; CS, control netting with a single SPB Insects Limited tab lure. State abbreviations: AR - Arkansas, KS - Kansas. There were three transects per site, each with every treatment above represented, thus a total of n = 8-9 replicate deployments in AR and n = 12 deployments in KS.,Trial 1 Recapture - To understand whether the method by which LLIN was deployed affected subsequent commodity infestation and progeny production, pilot-scale warehouses (5.85 × 2.81 m) in Manhattan, KS were used. At the far end of the warehouse against the back wall, a commodity consisting of a mixture of 210 mL organic, whole wheat kernels and 210 mL of organic, unbleached flour was placed. A total of 100 individuals each of T. castaneum, R. dominica, and T. variabile were released at the opposite end of the warehouse (approx. 5.25 m away). There were n = 12 replicate releases per treatment from 26 April 2019 to 16 August 2019, comprising a total of 3,600 released insects. There were four LLIN deployment methods that were tested (Figure 2). In the “hanging” treatment, LLIN (2.72 × 2.41 m) was affixed to the warehouse ceiling and allowed to hang down to the floor, completely bisecting the room. In the “cover” deployment method, LLIN was directly laid over the commodity. In the “pipe” deployment method, a PVC pipe (91 cm length, 5.1 cm I.D.) was bisected halfway with LLIN. These were compared with a "control" that used the same PVC pipe design, but without netting. Insects were given 72 h to disperse across the warehouse to the commodity. After this period, insects were collected by pre-designated zones in the warehouse. The zones were noted respective to the location of the commodity, and included "in commodity" (inside

,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,

,Trapping in 2023 with a linear set of dosages of (E)-8-dodecenyl acetate,Field trapping was done according to the methodology in Ruiz et al. 2022. The fields were located in North-Central Kansas at the Land Institute near Salina, KS. No pesticides were applied to these fields during the experiment in 2023. Starting the first week of June, six transects were set out, two in each Silphium integrifolium field. Each transect contained seven 30.4 cm x 30.4 cm sticky card traps (Alpha Scents, Canby, OR, USA) affixed to the top of a 1.27 cm diameter, three foot in length PVC pole that was hammered into the ground until sturdy. The cards were affixed using a 271 cm long sticky card ring holder (Olson Products Inc., Medina, OH, USA) that was bent to a 90° angle and placed inside the PVC pipe. Two large binder clips were also used to anchor the sticky card to its card holder.,The sticky traps in each transect were spaced 10 meters apart around the perimeter of the field. Within each transect, traps were baited with a linear increase in concentrations in 2023, including either a control (50 µl of acetone), a low concentration (50 µl of a solution made by mixing 5.75 µl of (E)-8-dodecenyl acetate in 5 ml of acetone), or a doubled concentration (11.5 µl of (E)-8-dodecenyl acetate diluted in 5 ml of acetone) of (E)-8-dodecenyl acetate (Alfa Chemistry, Ronkonkoma, NY, USA). All lures were added to a 3-ml LDPE dropping bottle (Wheaton, DWK Life Sciences, Millville, NJ, USA). The clear sticky card traps were collected and replaced biweekly until the first E. giganteana adult was caught, then traps were changed weekly. The lures and control bottles were replaced once every two weeks (with lure emissions confirmed out to 14 d in Ruiz et al. 2022) and their position in the field rotated at each change. Each lure was in each position twice over the course of the season.,When collected, the sticky cards were held in a 7.6 L (=2 gal) labeled Ziploc© bag transported back to USDA-ARS. All collected sticky traps were placed in a freezer for approximately 24 h. The total number of E. giganteana per trap and their distance from the lure in millimeters was recorded. In addition, the number of nontarget lepidoptera was recorded on each trap. Individual E. giganteana and non-target lepidoptera were only counted if more than half of the specimen was remaining on the sticky trap at the time of counting to ensure positive identification.,Trapping in 2024 with an exponential set of concentrations of (E)-8-dodecenyl acetate,Field trapping in 2024 was conducted similarly to that in 2023 with the following modifications. Three different fields located at the Land Institute were used (Table 1). [HS1] Pesticides were applied once to one of the fields and adjacent to one of the others. Three transects were deployed in each of the three fields. Each transect contained four traps for a total of 36 traps. The traps were assembled similarly to those used in 2023, but a hand-made sticky card was used instead of a manufactured one to improve captures. These sticky cards were made of a laminated 21.6 × 27.9 cm (=8.5 by 11 in) piece of white cardstock paper (Astrobright, Neenah, WI, USA) coated on both sides with TADⓇ all-weather adhesive (Trécé Adhesives Division, Adair, OK, USA). The sticky sides were covered with wax paper for ease of travel. Additionally, the sticky cards had a chicken wire cage placed over them in the field to try to prevent the capture of birds and other nontargets on the traps. Traps in 2024 were baited with an exponential set of concentrations of (E)-8-dodecenyl acetate. In each transect, there was a solvent only control (50 µl of acetone), a low concentration equivalent to the 2023 treatment (50 µl of a solution made of 5.75 µl of (E)-8-dodecenyl acetate diluted in 5 ml of acetone), a medium concentration (50 µl of a solution made of 78.5 µl of (E)-8-dodecenyl acetate diluted in 5 ml of acetone), and a high concentration (50 µl of a solution made

,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