Data from: Season-long microbial dynamics from the cuticle of rice weevil originating at food facilities after dispersal to novel food patches

<i>Trapping of field-collected </i>S. oryzaeTo capture field-collected<i> S. oryzae </i>to evaluate how microbial vectoring over the course of a season in 2022, six commercial pitfall traps (Storgard, Trécé Inc., Adair, OK, USA) were baited with 5 g of wheat and the <i>S. oryzae</i> aggregation pheromone, (4<i>S</i>,5<i>R</i>)-5-hydroxy-4-methyl-3-heptanone (IL-703, Insects Limited, Westfield, IN, USA) and deployed 10 m apart. In 2023, 14 pitfall traps (Storgard, Trécé Inc.) were deployed, separated by 5–15 m and baited with the same stimuli. In addition, probe traps (WB Probe II, Storgard, Trécé Inc., Adair, OK, USA) were taken in identical locations in adjacent grain bins, but &gt;90% of individuals came from the pitfall traps. Trapping took place at the Kansas State University Foundation Seed Farm (39°12'23"N, 96°35'42"W), and occurred on a weekly basis from 11 May 2022 to 28 Sep 2022 and 17 May 2023 to 1 Nov 2023. Live <i>S. oryzae</i> from each trap were placed in a sterile Ziplock bag and brought back to USDA-ARS Center for Grain and Animal Health in a cooler. Live weevils were stored at 23°C and 65% RH after brought back from the field. Within 24 h of being brought to the laboratory, <i>S. oryzae</i> were introduced to factitious, novel food patches as described below.<i>Linking with weather data</i>Weather data was obtained from the Kansas Mesonet system (https://mesonet.k-state.edu/) at a weather station located in the same location as the trapping on the Kansas State University Agronomy Farm (39°12'26"N, 96°35'42"W). Air temperature was measured at 1.98 m above ground level (HMP155 probe, Vaisala, Vantaa, Finland) inside of a non-aspirated radiation shield within an error range of ± 0.1°C. Data was acquired every hour with a microprocessor (CR3000 series, Campbell Scientific Inc., Logan, UT, USA), which accurately measures to the microvolt level and controls peripheral devices. Mean maximum and minimum temperature was compiled for each 7-day period preceding each date of collection for <i>S. oryzae</i> in 2022 and 2023, and it was linked to microbial growth measures detailed below from the same date.<i>Assessing microbial growth after dispersal to novel food patches</i>A total of n = 5 individual field-collected <i>S. oryzae</i> from each date were introduced individually onto petri dishes (100 ´ 20 mm) composed of potato dextrose agar as a factitious, novel food patch for 3 and 5 d to mimic dispersal (following the methods of Ponce et al. 2024). This was done within the confines of a permitted BSL2 (Permit# IBC-1693) space using a biosafety cabinet (75 × 73 × 95 cm L:H:W, #302381101, Labconco, Kansas City, MO, USA). After introduction of <i>S. oryzae</i>, the petri dishes were placed in an environmental chamber (Percival Scientific, Perry, IA, USA) set at constant conditions (30°C, 65% RH, and 14:10 L:D cycle). Petri dishes were photographed at 3 and 5 days of <i>S. oryzae </i>foraging in the novel food patch with a 3D-imaging system (Cognisys Inc., Traverse City, MI, USA) using a SLR camera (EOS 7D Mark II, Canon Inc., Tokyo, Japan) with a wide-angle lens (L series USM 17–40 mm, Canon Inc., Tokyo, Japan) and twin flash (MT-24X, Macro Twin Flash Lite, Canon Inc., Tokyo, Japan). Light was diffused using a partially cut frosted plastic jar (15.2 × 7.6 cm D:H). Images were processed using ImageJ v.1.53 (Wayne Rasband, National Institutes of Health, USA) individually by first subtracting the background, then finding edges, and converting the image to binary (white/black). As needed, erode and/or dilate was sparingly used to make sure the image reflected microbial growth in the original image. A circle encompassing only the Petri dish was created and the mean grayscale, standard deviation of the grayscale value, and count of pixels were measured as a surrogate for microbial growth on the dishes. This allowed a quantitative measure of microbial growth by creating an average in each image. The mean grayscale value could range from 0 (full white), indicating no microbial growth, to 255 (full black), indicating full microbial growth on the entire dish. Increased microbial growth was defined as higher mean grayscale values. Finally, visually, microbial morphospecies (alpha) richness was assigned to each image by two observers given the number of unique morphospecies on the plate as a proxy for community complexity. Where these numbers varied by observer (which was rare), both observers discussed together and came to a consensus on the number of morphospecies present in the petri dish. Microbial morphospecies have been prior used successfully to challenge the hypothesis “everything is everywhere, but, the environment selects” hypothesis (Telford et al. 2006).

野外采集米象的诱集
为评估2022年整个生长季内微生物介导的传带规律，我们设置了6个商用陷捕诱器（Storgard，Trécé Inc.，美国俄克拉荷马州阿达爾市），以5g小麦与米象（S. oryzae）聚集信息素(4S,5R)-5-羟基-4-甲基-3-庚酮（IL-703，Insects Limited，美国印第安纳州韦斯特菲尔德市）为诱饵，各诱器间距10米布置。2023年，我们部署了14个陷捕诱器（Storgard，Trécé Inc.），间距设置为5~15米，使用相同诱饵开展诱集。此外，我们在相邻粮库的相同位置布设了探针诱捕器（WB Probe II，Storgard，Trécé Inc.，美国俄克拉荷马州阿达爾市），但最终捕获的个体中超过90%均来自陷捕诱器。
本诱集试验于堪萨斯州立大学基金会种子农场（39°12'23"N，96°35'42"W）开展，采样周期为每周一次，时间覆盖2022年5月11日至2022年9月28日，以及2023年5月17日至2023年11月1日。将每个诱捕器捕获的活米象（S. oryzae）装入无菌Ziplock袋，置于冷藏箱中转运至美国农业部农业研究服务局（USDA-ARS）谷物与动物健康中心。带回实验室后，将活象鼻虫置于23℃、相对湿度65%的条件下保存。在野外采样带回实验室后的24小时内，按照下述方法将米象接种至人工制备的新型食物斑块中。

与气象数据关联
气象数据来源于堪萨斯州气象网络系统（Kansas Mesonet，https://mesonet.k-state.edu/），数据采集自与诱集试验同位置的堪萨斯州立大学农学农场气象站（39°12'26"N，96°35'42"W）。空气温度通过安装在非通风辐射屏蔽罩内的HMP155探头（Vaisala，芬兰万塔）在距地面1.98米处测定，测量误差范围为±0.1℃。数据通过微处理器（CR3000系列，Campbell Scientific Inc.，美国犹他州洛根市）每小时采集一次，该设备可精确测量微伏级信号并控制外围设备。我们分别统计了2022年和2023年每次采样日期前7天的平均最高、最低气温，并将其与下述同日获取的微生物生长测量数据进行关联分析。

评估新型食物斑块扩散后的微生物生长
我们从每次采样获得的野外采集米象中随机选取5头个体，将其分别接种至以马铃薯葡萄糖琼脂（PDA）为培养基的培养皿（100×20 mm）中，作为人工新型食物斑块，分别培养3天和5天以模拟昆虫扩散过程（参照Ponce等人2024年的方法）。所有操作均在获得许可的生物安全二级（BSL-2）实验室（许可编号IBC-1693）内的生物安全柜（75×73×95 cm 长×高×宽，#302381101，Labconco，美国密苏里州堪萨斯城）中完成。接种米象后，将培养皿置于环境培养箱（Percival Scientific，美国艾奥瓦州佩里市）中，设置恒定条件：30℃、相对湿度65%、光照周期14:10（L:D）。
在米象在新型食物斑块中觅食3天和5天后，使用3D成像系统（Cognisys Inc.，美国密歇根州特拉弗斯城）搭配单反相机（EOS 7D Mark II，佳能（Canon）公司，日本东京）、广角镜头（L系列USM 17–40 mm，佳能公司，日本东京）及双闪光灯（MT-24X，微距双闪光灯，佳能公司，日本东京）对培养皿进行拍照。光线通过部分裁剪的磨砂塑料罐（15.2×7.6 cm 直径×高度）进行漫射处理。
使用ImageJ v.1.53软件（Wayne Rasband，美国国立卫生研究院）对每张图像进行独立处理：首先扣除背景，随后提取边缘并将图像转换为二值图（白/黑模式）。根据需要少量使用腐蚀和/或膨胀操作，确保图像准确反映原图像中的微生物生长情况。绘制仅覆盖培养皿区域的圆形选区，测量该区域内的平均灰度值、灰度值标准差及像素总数，以此作为培养皿上微生物生长的替代指标，从而实现微生物生长的定量分析：平均灰度值的取值范围为0（全白，代表无微生物生长）至255（全黑，代表整个培养皿均被微生物覆盖），微生物生长量越高，平均灰度值也越高。
此外，由两名观察者分别对每张图像中的微生物形态种（alpha）丰富度进行赋值，以培养皿上的独特形态种数量作为群落复杂度的替代指标。若两名观察者的计数存在差异（这种情况极为罕见），则通过共同讨论达成一致意见，确定培养皿中的形态种数量。微生物形态种此前已被成功用于验证“万物皆可随处分布，但环境选择起主导作用”假说（Telford等人2006年）。