Data from: Adapting overwintering honey bee (<i>Apis mellifera</i> L.) colony management in response to warmer fall temperatures associated with climate change

The data represent estimates of honey bee colony sizes expressed as combs (frames) of adult bees and combs of brood (eggs, larvae and pupae). Colony sizes were estimated by dividing combs into one-tenth sections and counting the number covered with bees or brood on both sides of a comb. The values from each comb were summed and used as an estimate of combs with bees or brood for each colony. During pre-cold storage measurement in October, colonies were comprised of two deep Langstroth hive boxes. Temperatures were below 10^oC so the one-tenth of the comb area method for comb evaluations was not used. Instead, estimates of adult bee populations were made by tilting the top hive body forward on the pallet and frames with at least 75% bee coverage on the top and bottom of the comb were counted as a comb of bees. In the lower hive body, adult populations were estimated from adult bees covering the top of the combs. Combs with bees were totaled for the colony as an estimate of colony size. Estimates of Varroa mites (<i>Varroa destructor </i>Anderson &amp; Trueman) per 100 adult bees were made using approximately 300 worker bees per colony that were brushed from brood comb into jars containing 50 ml of 70% ethanol. Mites were counted by vigorously shaking the sample jars, and pouring the bees and alcohol into a strainer positioned over a pan. The mites that went through the strainer and into the pan were counted. Bees in the strainer also were examined for mites. All the bees in the sample were counted to estimate mites per 100 bees. Nosema (<i>Vairimorpha </i>(Microsporidia: Nosematidae) spore counts per colony were based on samples of 20 bees per colony. Samples were placed in a test tube containing ultra-pure water and homogenized for 5s. Samples equilibrated in 30-60 s, and the supernatant was removed from the center of the sample in the test tube (clear area) and placed into a 1.5ml Eppendorf tube. Spores per colony was estimated by transferring a 15 µl sample of supernatant to a hemocytometer, and examining it using a compound microscope at 400x with phase-contrast lighting. <i>Nosema</i> spores were counted in 16 small squares within five larger squares. The final spore count was calculated by multiplying the hemocytometer counts by 50,000. Fat body metrics (weight and lipid and protein concentrations) are based on pooled samples of 10 bees per colony. Fat bodies were removed by placing an adult worker bee onto a block of dry ice, and removing the abdomen. The entire gut was removed and the remaining abdominal carcass with fat body attached was rinsed to remove remaining gut contents and blotted dry. Fat body weight was estimated after drying the abdominal carcass at 60^oC for four days. Fat body protein concentration was estimated with a BCA Protein Assay kit. Samples were analyzed in triplicate and read in a microplate reader at a wave length of 562nm. The absorbance values of blank wells were subtracted from each standard and sample. Protein concentration (µg/µl) was estimated using the absorbance and standard curve. Lipid concentrations were estimated by placing the fat body sample into a tube containing a 2:1 mixture of chloroform:methanol (1 ml) along with 210 µl of 0.25% KCI. The sample was vortexed, and centrifuged at 2,000 rpm for 15 min. The bottom chloroform layer was removed and placed into a 2ml glass screw cap vial. Serial dilutions of corn oil dissolved in chloroform were prepared to construct a standard curve for lipid concentrations. The negative control consisted of 100 µl of chloroform. Samples, standards, and the negative control were dried to completion for approximately 1.5h. Dried samples were reacted with 182 µl of concentrated sulfuric acid at 100^oC for 15 min and 1478 µl of vanillin-phosphoric acid for 15 min in the dark at room temperature. Each of the negative controls, standards, and samples (100µl) were plated in triplicate and read using a spectrophotometer at 525nm. The average absorbance value for the negative control was subtracted from each standard and sample. A linear equation of the standards was derived to infer the µg/µl of lipid per sample from the absorbance values.本数据集为蜜蜂蜂群群势的估算值，以成年蜂巢脾（巢框，combs (frames)）和蜂子巢脾（含卵、幼虫及蛹，brood）的数量表示。群势估算的常规方法为：将单张巢脾划分为十分之一区域，计数巢脾两面覆盖成年蜂或蜂子的区域数量，累加所有巢脾的数值，作为对应蜂群的成年蜂或蜂子巢脾总数估算值。10月预冷藏期的测量中，受试蜂群采用两层深继箱朗氏蜂箱（Langstroth hive box）饲养，当时环境温度低于10℃，因此未采用巢脾十分之一区域评估法，改为通过以下方式估算成年蜂种群数量：将上层蜂箱在托盘上前倾，计数巢脾上下两面至少75%面积覆盖蜜蜂的巢框，计为1张成年蜂巢脾；下层蜂箱则以覆盖巢脾顶面的成年蜂数量估算种群规模，最终累加蜂群的成年蜂巢脾总数，作为该蜂群的群势估算值。每100只成年蜂的狄斯瓦螨（Varroa destructor Anderson & Trueman）寄生数量通过以下方法测定：每群采集约300只工蜂，从蜂子巢脾上刷入装有50ml 70%乙醇的样本瓶中，剧烈振荡样本瓶后，将蜜蜂与乙醇混合液倒入置于托盘上方的滤器中，收集通过滤器落入托盘的瓦螨，并检查滤器内残留的蜜蜂以统计剩余螨数，最后计数所有样本蜜蜂的总数量，据此计算每100只成年蜂的瓦螨寄生数量。每群的微粒子虫属（Vairimorpha (Microsporidia: Nosematidae)，原诺赛玛微孢子虫）孢子计数基于每群20只蜜蜂的样本：将样本置于装有超纯水的试管中均质5秒，静置30-60秒后，吸取试管内样本中央的澄清上清液至1.5ml艾本德离心管（Eppendorf tube）中，取15µl上清液滴加到血细胞计数板（hemocytometer）上，使用配备相差照明装置的复合显微镜在400倍视野下计数孢子数量，统计5个大方格内的16个小方格中的孢子总数，最终每群的孢子数通过将血细胞计数板的计数结果乘以50000得到。脂肪体相关指标（体重、脂质及蛋白质浓度）基于每群10只蜜蜂的混合样本：将成年工蜂置于干冰托盘上，摘除腹部并去除整个肠道，将附着脂肪体的腹部残体冲洗以去除残留肠道内容物，随后吸干水分，将腹部残体置于60℃环境下干燥4天后，称量得到脂肪体估算重量；脂肪体蛋白质浓度采用BCA蛋白定量试剂盒（BCA Protein Assay kit）检测，样本设置三次重复检测，在酶标仪（microplate reader）中于562nm波长下读取吸光度值，以空白孔的吸光度值校准标准品与待测样本的吸光度，通过吸光度值与标准曲线推算得到蛋白质浓度（单位：µg/µl）；脂质浓度检测流程为：将脂肪体样本置于装有2:1氯仿-甲醇混合液（1ml）和210µl 0.25%氯化钾溶液的试管中，涡旋混匀后以2000rpm离心15分钟，吸取下层氯仿层至2ml玻璃螺口样品瓶中，配制玉米油的氯仿梯度稀释液作为脂质浓度标准品，以100µl氯仿作为阴性对照，将样本、标准品及阴性对照各设置三次重复，置于环境中干燥约1.5小时，随后依次加入182µl浓硫酸，于100℃反应15分钟，再加入1478µl香草醛-磷酸溶液，室温避光反应15分钟，取100µl反应液在分光光度计（spectrophotometer）525nm波长下读取吸光度值，以阴性对照的平均吸光度值校准标准品与待测样本的吸光度，通过标准品的线性回归方程，由吸光度值推算得到每份样本的脂质浓度（单位：µg/µl）。