这项研究是由格兰特105AS-13.2.3-BQ-B1从动物局动植物卫生检验检疫，农业委员会，行政院和格兰特103-2313-B-197-002-MY3从支持部科学技术部（MOST）。

1.准备<ol><li>使1升50％的糖浆。溶解1公斤蔗糖在1L的DDH 2 O. </li><li>准备在BLD吡丙醚（PPN）的解决方案。使1.1升的10,000ppm的PPN原液，并稀释于无菌的DDH 2 O储存于4℃1 L 100毫升PPN溶液。 </li><li>稀释在BLD的PPN原液至0.1，1，10和100mg / kg的（PPM）最终浓度为下述实验。 </li><li>让PPN糖浆（为殖民地的水平）。稀释PPN股票的50％糖浆10和100ppm的最终浓度为以下实验。 </li><li>蜜蜂饲养。 注:在这里，实验地点是国立宜兰大学（NIU）养蜂场，艺兰市台; （GPS坐标:N24.747278，E121.746200）。 <ol><li>检查蜜蜂（ 意大利蜜蜂 L.）菌落每周食物量并在必要时（蜂蜜存储区是空的）用1升50％的糖浆进料。定义一个健康集落在每个集落，用大号产卵通常蜜蜂梳子的9个帧。 </li></ol></li><li>制备100毫升基本幼虫饮食（BLD）的。溶解6％D-葡萄糖，6％果糖，和1％的酵母提取物在无菌蒸馏水去离子水（DDH 2 O）和补充有50％蜂王浆。储存在4°C，但不超过3天。 注意:预温热至35℃实验前并在3天内使用。 </li></ol> 2. 体内送入方法注意: 体内喂养方法已经从汉利等改性。 20 <ol><li> 蜜蜂幼虫的选择和标签。 <ol><li>插入一个大号排阻到健康群体的9个帧分割成4个帧和5帧，同时限制王后到4帧区间。留在4帧部分的至少一个空帧产卵。 </li><li>蜂王产卵1天后，检查蛋的外观的帧并保持蜂箱72个小时（ 第 4天）内的鸡蛋直到1天老幼虫孵化。采用包含1日龄工人幼虫（24小时内画影线）出用手从测试配置单元中的帧中的一个和从与蜂刷框架拆除蜜蜂工人。 </li><li>覆盖有透明的滑动件（尺寸=长×宽×厚=29.7毫米×21毫米×0.1毫米）所述框架和钉对帧与图钉（ 图1A）的边缘上的透明幻灯片。 </li><li>对于每个处理，随机选择50一天的老幼虫（ 第 4天），并通过使用在透明滑动永久记号笔标记每个育雏细胞（ 图1A和1B）。 注意:通过使用永久记号笔，以避免不同的处理之间的混乱写帧和透明幻灯片上每个处理的信息，以及。除去标记的幻灯片，并保持用于体内&lt;/ em&gt;的进料和观察。 </li></ol></li><li> 体内喂养。 <ol><li>不同浓度的PPN-BLD（0.1，1，10和100ppm）的每个标记的育雏细胞，通过移液在1天，2和3加以分别为10μL，10μL和20μL，根据的吸气量幼虫龄。 BLD（无PPN）相同量添加到对照组。由此，PPN-BLD的每个标记的细胞育雏总剂量累积到4，40，400，和4000微克。 注意:通过标记透明载玻片识别标记的育雏细胞和喂食后除去标记透明幻灯片。使用新鲜BLD供给由工蜂防止育雏细胞的清洗。 </li><li>返回PPN处理帧到初始菌落作进一步的观察。 注:每个处理有四个殖民地4个生物重复。 </li></ol></li><li> 7天处理的幼虫观察。 <ol><li>观察日ËPPN处理幼虫，标记的透明滑动返回到帧以记录死亡率和封盖率标记的育雏细胞。 </li></ol></li><li> 13天治疗蛹羽化和利率的观察。 <ol><li>删除封顶育雏细胞的蜂蜡。 </li><li>把软尖镊子进入育雏细胞和夹紧蛹非常轻微地再取出来蛹轻轻。 </li><li>与实验室组织的双层下方各孔转移到蛹24孔组织培养板中。记录蛹转移过程中的损伤和死亡。 </li><li>保持在培养箱中在24孔组织培养板在34℃和70％相对湿度，直至出现（ 约 8天）。 </li><li>观察并记录蛹和出现的蜜蜂。 </li></ol></li><li> 统计 <ol><li>计算所记录的数据并呈现为平均值±SD </li><li>分析由SA使用方差分析（ANOVA）分析数据S和用最少的差异显著（LSD）检验，以分析不同处理两种手段的区别。定义为P值&lt;0.05具有统计学显著。不同的字母在表的同一列显示，通过统计分析显著的效果。 </li></ol></li></ol> 3. PPN的毒性在殖民地水平蜂箱<ol><li>成立蜜蜂群体。 <ol><li>插入一个大号排除器垂直于健康的群体的9个帧分割成4个帧（A部分）和5帧（B部分），同时限制王后到4帧区间。在用于产卵的4帧部分保留至少一个空帧。 注:换个女王逃生的女皇部分的顶部，以防止女王从零部件之间移动。 </li><li>女王铺设1天的鸡蛋后，检查帧鸡蛋的外观。用手取含有鸡蛋从部分A进行适当的帧并取出蜜蜂工人从与蜂刷帧。 </li><li>盖上透明滑动框架和钉透明幻灯片与图钉框架的边缘。 </li><li>随机地选择含有鸡蛋100个育雏细胞，并通过使用在透明幻灯片永久性记号笔标记每个育雏细胞。分配这些100个育雏细胞作为组1.收件使用永久标记，以避免不同的处理之间的混淆的帧和透明幻灯片上每个处理的信息。 </li><li>返回标记的帧部分用于3天，然后王后转移到B部分产卵。 </li><li> 1天后，检查部分B的帧中，选择一个合适的帧和标签如步骤3.1.4所述的含有鸡蛋100个育雏细胞。分配这些100个育雏细胞作为第2组。 </li><li>返回标记的帧到B部分3天，然后转移王后A部分产卵。 </li><li>反复A部分和B部分之间的女王交换6次以上和数值分配团，分别（ 图2）。应该有共9组。 </li></ol></li><li> ŤREAT蜂蜜蜂群与在第13天PPN糖浆（ 图2）。 <ol><li>加入1升含有在塑料蜂料箱10或100ppm（宽×长×高= 20厘米×30厘米×3.5厘米）的50％PPN糖浆和把盒子上在实验菌落的帧的顶部。 注:第1组没有13天内收到的PPN为育雏细胞已被查封。 </li><li>饲料对照组如步骤2.2中所述的1L 50％糖浆（无PPN）。 </li></ol></li><li>共1日龄幼体和记录作为每一组在第5天（ 图2）100个标记的育雏细胞的蛋孵化率。蜜蜂卵一般需要3天才能孵化成0天幼虫;因此，检查和标签100个鸡蛋含有育雏细胞在第1天，并在第5天计数1天老幼虫的数目对于每个组，以获得孵化率百分比。 </li><li> C指望。2-15封端的育雏细胞和记录作为每一组第11天（ 图2）100个标记的育雏细胞的幼虫封盖率。 6〜7日龄幼虫窝细胞会通过蜜蜂工人蜜蜂蜡幼虫化蛹为上限。 </li><li>观察蛹成熟和记录标记100个育雏细胞的羽化率对于每个组17天（ 图2）。 <ol><li>删除封顶育雏细胞的蜂蜡，并采取了蛹轻轻用软尖镊子。转移蛹于24孔组织培养板用实验室组织的双层之下每个孔中。 </li><li>保持在培养箱中在24孔组织培养板在34℃和70％RH，直到出现。 </li><li>观察并记录蛹和出现蜜蜂对于每个组，直到第9组（49实验天）。 注:每个处理有四个生物重复。 </li></ol></li><li>统计<ol><li>计算所记录的数据并呈现为平均±标准差。 </li><li>分析对治疗之间的显著差异（ 例如为0ppm / 10 ppm时，为10 ppm / 100ppm的和为0ppm / 100ppm的）每个组中通过使用学生双尾t检验。定义如果P值&lt;0.05为有统计学显著。 </li></ol></li></ol>

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W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Composition+of+freshly+harvested+and+commercial+royal+jelly.">Composition of freshly harvested and commercial royal jelly.</a> J. Apicult. Res. 24 (1), 52-61 (1985).</li></ol>

作者宣称，他们没有竞争的经济利益。

王后限制产蛋方法和王后交换法是用于获得本协议内的现场测试设置蜜蜂基团的关键步骤。王后限制产蛋方法允许蜜蜂的生命周期的同步。因此，研究人员可以选择相同的年龄与不同剂量农药的治疗1日龄幼虫。对于女王交换法，王后被部分A（4帧）和B（5帧）以获得蜜蜂的不同发育阶段的现场测试，以评估农药和农药残留的影响之间进行交换。此外，大量的选择育雏细胞被记录在使用透明幻灯片?...

农药在环境中的存在是最严重的问题之一是影响一个蜜蜂1，2，3的使用寿命。一些研究表明农药残留量的蜂蜜蜂群和蜜蜂产品的共同存在。在台湾，农药的平均应用为11-12公斤/公顷，每年（2005至13年）。农药在台湾使用量是比欧盟国家的高，以及拉丁美洲国家的4，5。换句话说，在养蜂环境正在遭受严重的农药压力，特别是在台湾，可能在其他国家。 在西方蜜蜂是农业系统6的主要传粉昆虫之一，它也产生有价值的产品，如蜂蜜。然而，蜜蜂是博览会ED各种杀虫剂和这些农药能觅食上采集花蜜和花粉7,8时已喷洒杀虫剂花后带回蜂箱。他们还可以通过自己的目的来控制荨麻疹9，10，11内的虫害问题养蜂人暴露于杀虫剂。由于蜜蜂幼虫被护士蜜蜂为他们的发展，幼虫，无人机喂，甚至女王可能会接触到这些农药污染的花蜜和花粉12。需要解决的13多种农药对蜜蜂的毒性。 许多已作出努力，以评估环境农药残留的问题。杨等人。测试的蜜蜂幼虫在发展中的神经毒性杀虫剂吡虫啉的影响蜂箱和报道，吡虫啉亚致死剂量导致成年蜂14的嗅觉关联行为。此外，Urlacher 等。检查的有机磷农药，毒死蜱，对蜜蜂工人的学习表现在实验室条件下15的亚致死效应。在我们以前的研究中，我们评估的昆虫生长调节剂（IGR）的影响，吡丙醚（PPN），幼虫蜜蜂16。 在本文中，我们提出，以评估对蜜蜂的发展化学影响的方法。进行了描述并应用于任何个别蜜蜂或菌落蜜蜂馈送方法。起初，我们在殖民地幼虫上测试了不同浓度的农药污染的基本幼虫饲料（BLD），以评估在体内个别蜜蜂农药的影响。我们接着来模拟自然康迪特通过使用蜂箱内农药污染的糖浆中的农药的离子。在该方法中，PPN，它被广泛用于对抗有害昆虫17和有害蜜蜂幼虫和蛹16，18，19的发展，将表示该领域的农药的负面影响的指标。

<table border="0" cellpadding="0" cellspacing="0" width="887">
	<tbody>
		<tr height="22">
			<td height="22" style="height:22px;width:193px;">Honey bee box</td>
			<td style="width:243px;">SAN-YI Honey Factory</td>
			<td style="width:155px;">W1266</td>
			<td style="width:297px;">Honeybees rearing</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:193px;">Queen excluder (between frames)</td>
			<td style="width:243px;">SAN-YI Honey Factory</td>
			<td style="width:155px;">I1575</td>
			<td style="width:297px;">Queen limitation&#160;</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:193px;">Queen excluder (on top )</td>
			<td style="width:243px;">SAN-YI Honey Factory</td>
			<td style="width:155px;">I1566</td>
			<td style="width:297px;">Queen limitation on top&#160;</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:193px;">Bee brush</td>
			<td style="width:243px;">SAN-YI Honey Factory, Taiwan</td>
			<td>W1414</td>
			<td style="width:297px;">clean the bees on frame gently</td>
		</tr>
		<tr height="18">
			<td height="18" style="height:18px;width:193px;">Bee feeder</td>
			<td style="width:243px;">SAN-YI Honey Factory, Taiwan</td>
			<td>P0219</td>
			<td style="width:297px;">feed sugar syrup to colony</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;">Transparent slide</td>
			<td style="width:243px;">Wan-Shih-Chei, Taiwan (http://www.mbsc.com.tw/a01goods.asp?s_id=40)</td>
			<td>1139</td>
			<td style="width:297px;">Mark the larval area on the frames (Material: Polyethylene Terephthalate, PET) (Size= Length*Width*thick= 29.7mm*21mm*0.1mm)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:193px;">24 well tissu culture plate</td>
			<td style="width:243px;">Guangzhou Jet Bio-Filtration Co., Ltd</td>
			<td style="width:155px;">TCP011024</td>
			<td style="width:297px;">Rearing pupae from extraction</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:193px;">Autoclave</td>
			<td style="width:243px;">Tomin medical equipmenco., LTD.</td>
			<td style="width:155px;">TM-321</td>
			<td style="width:297px;">Make sterilized distilled deionized water (ddH2O)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:193px;">P20 pipetman</td>
			<td style="width:243px;">Gilson</td>
			<td style="width:155px;">F123600</td>
			<td style="width:297px;">Add PPN into bee larval food pool</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:193px;">Incubator&#160;</td>
			<td style="width:243px;">Yihder Co., Ltd.</td>
			<td style="width:155px;">LE-550RD</td>
			<td style="width:297px;">Rearing pupae from extraction</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:193px;">Kimwipes</td>
			<td style="width:243px;">COW LUNG INSTRUMENT CO., LTD</td>
			<td style="width:155px;">KCS34155</td>
			<td style="width:297px;">Rearing pupae from extraction</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:193px;">Royal jelly</td>
			<td style="width:243px;">National Ilan University (NIU)</td>
			<td style="width:155px;">NIU</td>
			<td style="width:297px;">Make basic larval diet (BLD)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:193px;">D-(+)-Glucose</td>
			<td style="width:243px;">Sigma</td>
			<td style="width:155px;">G8270</td>
			<td style="width:297px;">Make basic larval diet (BLD)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:193px;">D-(-)-Fructose</td>
			<td style="width:243px;">Sigma</td>
			<td style="width:155px;">F0127</td>
			<td style="width:297px;">Make basic larval diet (BLD)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:193px;">Yeast extract</td>
			<td style="width:243px;">CONDA, pronadisa</td>
			<td style="width:155px;">1702</td>
			<td style="width:297px;">Make basic larval diet (BLD)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:193px;">Sucrose</td>
			<td style="width:243px;">Taiwan sugar coporation</td>
			<td style="width:155px;">E01071010</td>
			<td style="width:297px;">Make sugar syrup for bee food</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:193px;">Pyriproxyfen (11%)</td>
			<td style="width:243px;">LIH-NUNG CHEMICAL CO.. LTD.</td>
			<td style="width:155px;">Registration No. 1937</td>
			<td style="width:297px;">Insect growth regulator (IGR) used in the experiment</td>
		</tr>
	</tbody>
</table>

evaluating the effect of environmental chemicals on honey bee development from the individual to colony level

The presence of pesticides in the beekeeping environment is one of the most serious problems that impacts the life of a honey bee. Pesticides can be brought back to the beehive after the bees have foraged on flowers that have been sprayed with pesticides. Pesticide contaminated food can be exchanged between workers which then feed larvae and therefore can potentially affect the development of honey bees. Thus, residual pesticides in the environment can become a chronic damaging factor to honey bee populations and gradually lead to colony collapse. In the presented protocol, honey bee feeding methods are described and applied to either an individual honey bee or to a colony. Here, the insect growth regulator (IGR) pyriproxyfen (PPN), which is widely used to control pest insects and is harmful to the development of honey bee larvae and pupae, is used as the pesticide. The presenting procedure can be applied to other potentially harmful chemicals or honeybee pathogens for further studies.

为蜜蜂现场测试中，大号仅限于4帧部分用于产卵。此步骤可能会增加育雏密度在一帧和促进随后的观测。每个处理标记，和蜜蜂的发展显然通过透明幻灯片观察。 在 PPN-BLD的体内料到在蜂箱蜜蜂幼虫进行精确评估PPN对蜜蜂的殖民地发展的影响。使用体内供给方法促进了在蜂箱的化学处理的影响的观察。 对?...

Watch this Scientific Journal Video about Evaluating the Effect of Environmental Chemicals on Honey Bee Development from the Individual to Colony Level at JoVE.com

Evaluating the Effect of Environmental Chemicals on Honey Bee Development from the Individual to Colony Level

在这里我们提出 的方法，以农药污染的食物输送到两个单独的蜜蜂和蜂箱菌落。过程评估由基本幼虫饮食的体内喂养和也对蜂窝菌落的天然条件对个体蜜蜂的农药效果。

从个体到群体度评价环境化学物质对蜜蜂发展的影响

The presence of pesticides in the beekeeping environment is one of the most serious problems that impacts the life of a honey bee. Pesticides can be ...

evaluating-effect-environmental-chemicals-on-honey-bee-development

Research

JoVE Journal

Environment

9.0K 次观看.  National Ilan University. 在这里我们提出 的方法，以农药污染的食物输送到两个单独的蜜蜂和蜂箱菌落。过程评估由基本幼虫饮食的体内喂养和也对蜂窝菌落的天然条件对个体蜜蜂的农药效果。 

视频: Evaluating the Effect of Environmental Chemicals on Honey Bee Development from the Individual to Colony Level

Automated Gel Size Selection to Improve the Quality of Next-generation Sequencing Libraries Prepared from Environmental Water Samples

Next-generation sequencing of environmental samples can be challenging because of the variable DNA quantity and quality in these samples. High quality DNA libraries are needed for optimal results from next-generation sequencing. Environmental samples such as water may have low quality and quantities of DNA as well as contaminants that co-precipitate with DNA. The mechanical and enzymatic processes involved in extraction and library preparation may further damage the DNA. Gel size selection enables purification and recovery of DNA fragments of a defined size for sequencing applications. Nevertheless, this task is one of the most time-consuming steps in the DNA library preparation workflow. The protocol described here enables complete automation of agarose gel loading, electrophoretic analysis, and recovery of targeted DNA fragments. 
In this study, we describe a high-throughput approach to prepare high quality DNA libraries from freshwater samples that can be applied also to other environmental samples. We used an indirect approach to concentrate bacterial cells from environmental freshwater samples; DNA was extracted using a commercially available DNA extraction kit, and DNA libraries were prepared using a commercial transposon-based protocol. DNA fragments of 500 to 800 bp were gel size selected using Ranger Technology, an automated electrophoresis workstation. Sequencing of the size-selected DNA libraries demonstrated significant improvements to read length and quality of the sequencing reads.

This manuscript describes an automated gel size selection approach for purifying DNA fragments for next-generation sequencing. The Ranger Technology provides complete automation of the entire process of agarose gel loading, electrophoretic analysis, and recovery of targeted DNA fragments allowing for high-throughput and high quality next-generation sequencing libraries.

自动凝胶尺寸选择，以提高下一代测序文库的质量，从环境水样准备

Next-generation sequencing of environmental samples can be challenging because of the variable DNA quantity and quality in these samples. High quality DNA ...

科研

环境科学

Exploring the Effects of Atmospheric Forcings on Evaporation: Experimental Integration of the Atmospheric Boundary Layer and Shallow Subsurface

Evaporation is directly influenced by the interactions between the atmosphere, land surface and soil subsurface. This work aims to experimentally study evaporation under various surface boundary conditions to improve our current understanding and characterization of this multiphase phenomenon as well as to validate numerical heat and mass transfer theories that couple Navier-Stokes flow in the atmosphere and Darcian flow in the porous media. Experimental data were collected using a unique soil tank apparatus interfaced with a small climate controlled wind tunnel. The experimental apparatus was instrumented with a suite of state of the art sensor technologies for the continuous and autonomous collection of soil moisture, soil thermal properties, soil and air temperature, relative humidity, and wind speed. This experimental apparatus can be used to generate data under well controlled boundary conditions, allowing for better control and gathering of accurate data at scales of interest not feasible in the field. Induced airflow at several distinct wind speeds over the soil surface resulted in unique behavior of heat and mass transfer during the different evaporative stages.

A protocol for the design and construction of a soil tank interfaced to a small climate controlled wind tunnel to study the effects of atmospheric forcings on evaporation is presented. Both the soil tank and wind tunnel are instrumented with sensor technologies for the continuous in situ measurement of environmental conditions.

在探索大气蒸发强迫的影响:大气边界层和浅地层的实验整合

Evaporation is directly influenced by the interactions between the atmosphere, land surface and soil subsurface. This work aims to experimentally study ...

Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability

In situ recovery (ISR) is the predominant method of uranium extraction in the United States. During ISR, uranium is leached from an ore body and extracted through ion exchange. The resultant production bleed water (PBW) contains contaminants such as arsenic and other heavy metals. Samples of PBW from an active ISR uranium facility were treated with cupric oxide nanoparticles (CuO-NPs). CuO-NP treatment of PBW reduced priority contaminants, including arsenic, selenium, uranium, and vanadium. Untreated and CuO-NP treated PBW was used as the liquid component of the cell growth media and changes in viability were determined by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay in human embryonic kidney (HEK 293) and human hepatocellular carcinoma (Hep G2) cells. CuO-NP treatment was associated with improved HEK and HEP cell viability. Limitations of this method include dilution of the PBW by growth media components and during osmolality adjustment as well as necessary pH adjustment. This method is limited in its wider context due to dilution effects and changes in the pH of the PBW which is traditionally slightly acidic&#160;however; this method could have a broader use assessing CuO-NP treatment in more neutral waters.

Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability

Production bleed water (PBW) was treated with cupric oxide nanoparticles (CuO-NPs) and cellular toxicity was assessed in cultured human cells. The goal of this protocol was to integrate the native environmental sample into a cell culture format assessing the changes in toxicity due to CuO-NP treatment.

去除微量元素的氧化铜纳米粉体制备铀<em&gt;原位</em&gt;恢复泌水及其对细胞活力的影响

In situ recovery (ISR) is the predominant method of uranium extraction in the United States. During ISR, uranium is leached from an ore body and extracted ...

A Flow-through Exposure System for Evaluating Suspended Sediments Effects on Aquatic Life

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory by using pumps and automating delivery of sediment and water to simulate suspension of sediment. FLEES data are used to develop exposure-response curves between the effects on aquatic organisms and suspended sediment concentrations at the desired exposure duration. The effects data are used to evaluate management practices used to reduce the interactions between aquatic organisms and anthropogenic causes of suspended sediments. The FLEES is capable of generating total suspended solids (TSS) concentrations as low as 30 to as high as 800 mg/L, making this system an ideal choice for evaluating the effects of TSS resulting from many activities including simulating low ambient levels of TSS to evaluating sources of suspended sediments from dredging operations, vessel traffic, freshets, and storms.

A robust and flexible flow-through exposure system designed to maintain sediment in suspension is presented. The system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory.

对水生生命值评价悬浮泥沙影响的流通系统曝光

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended ...

Extraction of Organochlorine Pesticides from Plastic Pellets and Plastic Type Analysis

Plastic resin pellets, categorized as microplastics (&#8804;5 mm in diameter), are small granules that can be unintentionally released to the environment during manufacturing and transport. Because of their environmental persistence, they are widely distributed in the oceans and on beaches all over the world. They can act as a vector of potentially toxic organic compounds (e.g., polychlorinated biphenyls) and might consequently negatively affect marine organisms. Their possible impacts along the food chain are not yet well understood. In order to assess the hazards associated with the occurrence of plastic pellets in the marine environment, it is necessary to develop methodologies that allow for rapid determination of associated organic contaminant levels. The present protocol describes the different steps required for sampling resin pellets, analyzing adsorbed organochlorine pesticides (OCPs) and identifying the plastic type. The focus is on the extraction of OCPs from plastic pellets by means of a pressurized fluid extractor (PFE) and on the polymer chemical analysis applying Fourier Transform-InfraRed (FT-IR) spectroscopy. The developed methodology focuses on 11 OCPs and related compounds, including dichlorodiphenyltrichloroethane (DDT) and its two main metabolites, lindane and two production isomers, as well as the two biologically active isomers of technical endosulfan. This protocol constitutes a simple and rapid alternative to existing methodology for evaluating the concentration of organic contaminants adsorbed on plastic pieces.

Microplastics act as vector of potentially toxic organic contaminants with unpredictable effects. This protocol describes an alternative methodology for assessing the levels of organochlorine pesticides adsorbed on plastic pellets and identifying the polymer chemical structure. The focus is on pressurized fluid extraction and attenuated total reflectance Fourier transform infrared spectroscopy.

从塑料颗粒中提取有机氯农药和塑料类型分析

Plastic resin pellets, categorized as microplastics (&#8804;5 mm in diameter), are small granules that can be unintentionally released to the environment ...

Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation

Dimethylated thioarsenicals such as dimethylmonothioarsinic acid (DMMTAV) and dimethyldithioarsinic acid (DMDTAV), which are produced by the metabolic pathway of dimethylarsinic acid (DMAV) thiolation, have been recently found in the environment as well as human organs. DMMTAV and DMDTAV can be quantified to determine the ecological effects of dimethylated thioarsenicals and their stability in environmental media. The synthesis method for these compounds is unstandardized, making replicating previous studies challenging. Furthermore, there is a lack of information about storage techniques, including storage of compounds without species transformation. Moreover, because only limited information about synthesis methods is available, there may be experimental difficulties in synthesizing standard chemicals and performing quantitative analysis. The protocol presented herein provides a practically modified synthesis method for the dimethylated thioarsenicals, DMMTAV and DMDTAV, and will help in the quantification of species separation analysis using high performance liquid chromatography in conjunction with inductively coupled plasma mass spectrometry (HPLC-ICP-MS). The experimental steps of this procedure were modified by focusing on the preparation of chemical reagents, filtration methods, and storage.

Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation

This article presents modified experimental protocols for dimethylmonothioarsinic acid (DMMTAV) and dimethyldithioarsinic acid (DMDTAV) synthesis, inducing dimethylarsinic acid (DMAV) thiolation through mixing of DMAV, Na2S, and H2SO4. The modified protocol provides an experimental guideline, thereby overcoming limitations of the synthesis steps that could have caused experimental failures in quantitative analysis.

使用 DMAv为环境应用 DMMTAv和 DMDTAv的准备: 合成、纯化和确认

Dimethylated thioarsenicals such as dimethylmonothioarsinic acid (DMMTAV) and dimethyldithioarsinic acid (DMDTAV), which are produced by the metabolic ...

The Effect of the Application of Thyme Essential Oil on Microbial Load During Meat Drying

Meat is a high protein meal that is used in the preparation of jerky, a popular food snack, where preservation and safety are important. To assure food safety and to extend the shelf life of meat and meat products, the use of either synthetic or natural preservatives have been applied to control and eliminate foodborne bacteria. A growing interest in the application of natural food additives for meat has increased. Microorganisms, such as Escherichia coli, contaminate meat and meat products, causing foodborne illnesses. Therefore, it is necessary to improve the meat conservation process. However, the use of essential oils when the meat is being dried has not been deeply studied. In this regard, there is an opportunity to increase the value of dried meat and reduce the risk of foodborne illnesses by applying essential oils during the drying process. In this protocol, we present a novel method of applying thyme essential oil (TEO) during meat drying, specifically in vapor form directly in a drying chamber. For evaluation, we use Minimal Inhibitory Concentration (MIC) to detect the number of harmful bacteria in the treated samples compared to raw samples. The preliminary results show that this method is a viable and alternative option to synthetic preservatives and that it significantly reduces microbial load in dried meat.

Microorganisms such as Escherichia coli that contaminate meat products cause foodborne illnesses. The use of essential oils in the meat drying process has not been deeply studied. Here, we present a novel method of applying thyme essential oil to meat during drying to reduce the microbial load in dried meat.

百里香精油应用于肉类干燥过程中微生物负荷的影响

Meat is a high protein meal that is used in the preparation of jerky, a popular food snack, where preservation and safety are important. To assure food ...

Protocol for Acute and Chronic Ecotoxicity Testing of the Turquoise Killifish Nothobranchius furzeri

The killifish Nothobranchius furzeri is an emerging model organism in the field of ecotoxicology and its applicability in acute and chronic ecotoxicity testing has been demonstrated. Overall, the sensitivity of the species to toxic compounds is in the range with, or higher than, that of other model species.
This work describes protocols for acute, chronic, and multigenerational bioassays of single and combined stressor effects on N. furzeri. Due to its short maturation time and life-cycle, this vertebrate model allows the study of endpoints such as maturation time and fecundity within four months. Transgenerational full life-cycle exposure trials can be performed in as little as 8 months. Since this species produces eggs that are drought-resistant and remain viable for years, the on-site culture of the species is not needed but individuals can be recruited when required. The protocols are designed to measure life-history traits (mortality, growth, fecundity, weight) and critical thermal maximum.

Protocol for Acute and Chronic Ecotoxicity Testing of the Turquoise Killifish Nothobranchius furzeri

In this work, we describe an acute, chronic and multigenerational bioassay to study the effects of single and combined stressors on the Turquoise killifish Nothobranchius furzeri. This protocol is designed to study life-history traits (mortality, growth, fecundity, weight) and critical thermal maximum.

绿松石鳉鱼的急性和慢性毒性测试协议Nothobranchius furzeri

The killifish Nothobranchius furzeri is an emerging model organism in the field of ecotoxicology and its applicability in acute and chronic ecotoxicity ...

Measuring Hypopharyngeal Gland Acinus Size in Honey Bee (Apis mellifera) Workers

The nurse hypopharyngeal glands produce the protein fraction of the worker and royal jelly that is fed to developing larvae and queens. These paired glands that are located in the head of the bee are highly sensitive to the quantity and quality of pollen and pollen substitutes that the nurse bee consumes. The glands get smaller when nurses are fed deficient diets and are large when they are fed complete diets. Because nurse hypopharyngeal gland size is a robust indicator of nurse nutrition, it is essential that those studying honey bee nutrition know how to measure these glands. Here, we provide detailed methods for dissecting, staining, imaging, and measuring nurse bee hypopharyngeal glands. We present comparisons of unstained and stained tissue and data that were used to study the impact of pollen on gland size. This method has been used to test how diet impacts hypopharyngeal gland size but has further use for understanding the role of these glands in hive health.

Measuring Hypopharyngeal Gland Acinus Size in Honey Bee Apis mellifera Workers

Hypopharyngeal gland acinus size is a robust measure of nurse honey bee nutrition. Here, we provide a detailed protocol for dissecting, staining, imaging, and measuring nurse bee hypopharyngeal gland acini.

蜂蜜蜂 (api 蜜蜂) 中下咽腺性腺泡大小的测定

The nurse hypopharyngeal glands produce the protein fraction of the worker and royal jelly that is fed to developing larvae and queens. These paired ...

Composition and Distribution Analysis of Bioaerosols Under Different Environmental Conditions

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, we described a protocol for multiple analyses of the biological compositions in environmental PM. Five experiments are presented: (1) PM number monitoring by using a laser particle counter; (2) PM collection by using a cyclonic aerosol sampler; (3) PM collection by using a high-volume air sampler with filters; (4) culturable microbes collection by the Andersen six-stage sampler; and (5) detection of biological composition of environmental PM by bacterial 16SrDNA and fungal ITS region sequencing. We selected hazy days and a livestock farm as two typical examples of the application in this protocol. In this study, these two sampling methods, cyclonic aerosol sampler and filter sampler, showed different sampling efficiency. The cyclonic aerosol sampler performed much better in terms of collecting bacteria, while these two methods showed the same efficiency in collecting fungi. Filter samplers can work under low temperature conditions while cyclonic aerosol samplers have a sampling limitation for temperature. A solid impacting sampler, such as an Andersen six-stage sampler, can be used to sample bioaerosols directly into the culture medium, which increases the survival rate of culturable microorganisms. However, this method mainly relies on culture while more than 99% of microbes cannot be cultured. DNA extracted from the culturable bacteria collected by the Andersen six-stage sampler and samples collected by cyclonic aerosol sampler and filter sampler were detected by bacterial 16S rDNA and fungal ITS region sequencing.All the methods above may have wide application in many fields of study, such as environmental monitoring and airborne pathogen detection. From these results, we can conclude that these methods can be used under different conditions and may help other researchers further explore the health impacts of environmental bioaerosols.

Understanding the biological composition of environmental particulate matter is important for the study of its significant impacts on human health and disease spread. Here, we used three types of bioaerosol sampling methods and a biological analysis of airborne microbes to better explore airborne microbial communities under different environmental conditions.

不同环境条件下生物气溶胶的组成及分布分析

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, ...