Data from: Direct lethality and time-delayed sublethal effects of multiple types of insecticide netting against stored product insects

<i>Experimental Insects</i>The field strains of <i>T. castaneum</i> and <i>R. dominica</i> (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.<i>Treatments</i>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.<i>Direct Lethality Assessments</i>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.<i>Baseline Mobility Assay after Exposure to LLINs</i>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.<i>Comparison of Sublethal Effects among LLINs</i>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 affected as defined in Ranabhat et al. (2022) were used in this assay. In total, n = 16–20 replicates were performed per treatment combination.

「实验用昆虫」
本研究使用了野外品系的赤拟谷盗（*Tribolium castaneum*）与谷蠹（*Rhyzopertha dominica* (F.)）。前者于2012年采自堪萨斯州东部，后者同样采自堪萨斯州东部，但采集时间为2019年。所有供试物种均采用4至8周龄的成虫。饲养工作在美国农业部堪萨斯州曼哈顿市的谷物与动物健康研究中心（USDA Center for Grain Animal Health Research）完成。赤拟谷盗的饲养饲料为95%未漂白有机面粉与5%啤酒酵母的混合物，谷蠹则饲养于调质有机全麦谷物中。种群维持条件设置为温度27.5℃、相对湿度65%，光周期为14:10或16:8（光照:黑暗）小时。

「处理组设置」
处理组包括三种不同类型的长效杀虫蚊帐（long-lasting insecticide-incorporated netting, LLIN），具体如下：1）Carifend®：含0.34%高效氯氰菊酯的LLIN（40旦尼尔，BASF，路德维希港，德国）；2）D-Terrence：含0.4%溴氰菊酯的LLIN（2×2 mm网孔，Vestergaard SA，洛桑，瑞士）；3）8%醚菊酯LLIN（AgBio, Inc，科罗拉多州，美国）；对照组则使用与Carifend或Vestergaard同款但不含杀虫剂的蚊帐。

「直接致死性评估」
将20头混合性别的成虫组暴露于固定于9×9 cm²培养皿中的蚊帐上，暴露时长分别为5、60或120分钟。暴露结束后，分别在0、24、72或168小时后评估试虫状态（参照Ranabhat等（2022）的定义，将试虫分为存活、受影响或死亡三类），评估所用培养皿不含蚊帐，底部铺有直径8.5 cm的滤纸。简言之，存活成虫定义为可正常移动、活动，且翻转后能够自行扶正；受影响成虫表现为行动迟缓或步态不稳，翻转后无法自行扶正，部分或全部肢体出现抽搐；死亡成虫则完全无法移动。暴露后的试虫置于与种群饲养相同的环境条件下，但不提供补充饲料。每个物种的每个处理组合均设置n=4次重复。

「长效杀虫蚊帐暴露后的基线活动性测定」
基于致死性评估的观察结果，本研究的基线活动性测定聚焦于Carifend®与D-Terrence两种LLIN。仅使用存活成虫，在6个单独的培养皿（100×15 mm 直径:高度）中评估其活动性，培养皿底部铺有直径85 mm的1号滤纸（GE Healthcare，白金汉郡，英国）。处理组包括阴性对照（仅滤纸）、两种LLIN之一，以及与Carifend或Vestergaard同款但不含杀虫剂的蚊帐（作为阳性对照）。使用安装在培养皿上方80 cm处的网络摄像机（GigE，Basler AG，埃伦堡，德国）追踪其60分钟内的活动情况。培养皿后方使用LED灯箱（42×30 cm 宽:长，LPB3型号，Litup，深圳，中国）进行背光照明以提升对比度，并用白色泡沫板固定位置。视频流传输至计算机，使用Ethovision软件（v.14.0，Noldus Inc，利斯堡，弗吉尼亚州）进行分析。该程序可自动计算每头成虫在60分钟内的总移动距离（cm）与瞬时移动速度（cm/s）。每头成虫仅使用一次，视为一个重复。每个处理组合共计设置n=18次重复。

「长效杀虫蚊帐间亚致死效应的比较」
在亚致死活动性测定中，将混合性别的成虫暴露于Carifend®、D-Terrence LLIN或上述对照蚊帐中。将5~10头成虫组成的组暴露于固定于9×9 cm²培养皿中的LLIN上，暴露时长为5或60分钟。暴露结束后，立即或在72小时后，在与种群饲养相同的环境条件下（不提供补充饲料）的培养皿中评估LLIN对成虫活动性的影响，随后使用上述视频追踪系统与Ethovision软件进行分析。本测定仅使用经Ranabhat等（2022）定义为存活或受影响的试虫。每个处理组合共计设置n=16~20次重复。