Name: protocol for acute and chronic ecotoxicity testing of the turquoise killifish nothobranchius furzeri
Uploaded: 2018-04-24T14:30:00
Description: Watch this Scientific Journal Video about Protocol for Acute and Chronic Ecotoxicity Testing of the Turquoise Killifish Nothobranchius furzeri at JoVE.com

我们感谢 UAntwerpen 的球体小组和 Ugent 的作物保护部对水样进行分析。该项目的支持是由 PF/10/007 研究基金的卓越中心的生态和社会进化动态提供的。afg 胶 (11Q0516N) 和 ESJT (FWO-SB151323) 以博士和 TP (12F0716N) 为博士后研究员, FWO 佛兰德 (全宗 Wetenschappelijk Onderzoek)。

这里描述的所有方法都得到了 KULeuven 伦理委员会的批准。
1. furzeri的孵化和一般维护
<ol><li>准备鱼培养基 (pH 值 7), 温度为14摄氏度, 并加入纯化型 II 水, 加标准化盐, 电导率为600µS/厘米 (24 摄氏度)。</li>
	<li>从已存储在标准条件13下的 GRZ 实验室线中选择鸡蛋。选择 DIII 阶段的鸡蛋 (即准备孵化), 可识别的金色眼睛9 , 并轻轻地?...

<ol>
	<li>European-Chemicals-Bureau. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> TAPIR Three point three-A Project for the Information Requirements of REACH. Final Report-2 August 2005. Scoping study on the development of a Technical Guidance Document on information requirements on intrinsic properties of substances (RIP 3.3-1). , (2005).</li><li>Philippe, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+and+chronic+sensitivity+to+copper+of+a+promising+ecotoxicological+model+species,+the+annual+killifish+Nothobranchius+furzeri.">Acute and chronic sensitivity to copper of a promising ecotoxicological model species, the annual killifish Nothobranchius furzeri.</a> Ecotoxicol Environ Saf. , 26-35 (2017).</li><li>Noyes, P. D., Lema, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Forecasting+the+impacts+of+chemical+pollution+and+climate+change+interactions+on+the+health+of+wildlife.">Forecasting the impacts of chemical pollution and climate change interactions on the health of wildlife.</a> Current Zoology. 61 (4), 669-689 (2015).</li><li>Consommateurs, S. S. d. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Health &amp; Consumer Protection Directorate-General European Commission. 4, (2002).</li><li>Commission, E. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Guidance+document+on+aquatic+ecotoxicology.+Under+Council+directive+91/414/EEC.">Guidance document on aquatic ecotoxicology. Under Council directive 91/414/EEC.</a> SANCO/3268/2001 Rev 4. 2002b. , (2002).</li><li>. Ecological Effects Test Guidelines, OPPTS 850.1500 Fish life cycle toxicity Available from: <a target="_blank" href="https://www.epa.gov/sites/production/files/2015-07/documents/850-1500.pdf">https://www.epa.gov/sites/production/files/2015-07/documents/850-1500.pdf</a> (1996)</li><li>Philippe, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+and+chronic+sensitivity+to+copper+of+a+promising+ecotoxicological+model+species,+the+annual+killifish+Nothobranchius+furzeri.">Acute and chronic sensitivity to copper of a promising ecotoxicological model species, the annual killifish Nothobranchius furzeri.</a> Ecotoxicol Environ Saf. , (2017).</li><li>Ankley, G. T., Villeneuve, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+fathead+minnow+in+aquatic+toxicology:+past,+present+and+future.">The fathead minnow in aquatic toxicology: past, present and future.</a> Aquatic Toxicology. 78 (1), 91-102 (2006).</li><li>Pola&#269;ik, M., Bla&#382;ek, R., Reichard, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laboratory+breeding+of+the+short-lived+annual+killifish+Nothobranchius+furzeri.">Laboratory breeding of the short-lived annual killifish Nothobranchius furzeri.</a> Nature Protocols. 11 (8), 1396-1413 (2016).</li><li>Reichwald, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insights+into+Sex+Chromosome+Evolution+and+Aging+from+the+Genome+of+a+Short-Lived+Fish.">Insights into Sex Chromosome Evolution and Aging from the Genome of a Short-Lived Fish.</a> Cell. 163 (6), 1527-1538 (2015).</li><li>Valenzano, D. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+African+Turquoise+Killifish+Genome+Provides+Insights+into+Evolution+and+Genetic+Architecture+of+Lifespan.">The African Turquoise Killifish Genome Provides Insights into Evolution and Genetic Architecture of Lifespan.</a> Cell. 163 (6), 1539-1554 (2015).</li><li>Shedd, T. R., Widder, M. W., Toussaint, M. W., Sunkel, M. C., Hull, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+the+annual+killifish+Nothobranchius+guentheri+as+a+tool+for+rapid+acute+toxicity+screening.">Evaluation of the annual killifish Nothobranchius guentheri as a tool for rapid acute toxicity screening.</a> Environ. Toxicol. Chem. 18 (10), 2258-2261 (1999).</li><li>Platzer, M., Englert, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nothobranchius+furzeri:+a+model+for+aging+research+and+more.">Nothobranchius furzeri: a model for aging research and more.</a> Trends Genet. 32 (9), 543-552 (2016).</li><li>Watters, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+ecology+and+distribution+of+Nothobranchius+fishes.">The ecology and distribution of Nothobranchius fishes.</a> J Am Killifish Assoc. 42, 58-61 (2009).</li><li>Op de Beeck, L., Verheyen, J., Stoks, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Competition+magnifies+the+impact+of+a+pesticide+in+a+warming+world+by+reducing+heat+tolerance+and+increasing+autotomy.">Competition magnifies the impact of a pesticide in a warming world by reducing heat tolerance and increasing autotomy.</a> Environ Pollut. 233, 226-234 (2018).</li><li>Patra, R. W., Chapman, J. C., Lim, R. P., Gehrke, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+three+organic+chemicals+on+the+upper+thermal+tolerances+of+four+freshwater+fishes.">The effects of three organic chemicals on the upper thermal tolerances of four freshwater fishes.</a> Environ. Toxicol. Chem. 26 (7), 1454-1459 (2007).</li><li>Beitinger, T. L., Bennett, W. A., McCauley, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temperature+tolerances+of+North+American+freshwater+fishes+exposed+to+dynamic+changes+in+temperature.">Temperature tolerances of North American freshwater fishes exposed to dynamic changes in temperature.</a> Environ Biol Fishes. 58 (3), 237-275 (2000).</li><li>Cellerino, A., Valenzano, D. R., Reichard, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+the+bush+to+the+bench:+the+annual+Nothobranchius+fishes+as+a+new+model+system+in+biology.">From the bush to the bench: the annual Nothobranchius fishes as a new model system in biology.</a> Biological Reviews. , (2015).</li></ol>

这项工作描述了一种新的生物测定方法, 使用Nothobranchius furzeri(一种新兴的模型有机体) 来研究毒物和其他压力源的个体和组合的长期效应。提出的协议已成功地应用于测量物种对一系列有毒物质 (铜、镉、34-dichloroaniline 和毒死蜱) 的敏感性。由于它的快速生命周期, 这个脊椎动物模型允许评估致死和跨代的影响在四月内。使用这种鱼类作为毒性筛选模型的另一个主要优点是它生产抗旱蛋。这...

将一组物种的敏感度配置为战略性地选定的毒物被描述为欧洲范围立法 (注册、评估、授权和限制化学品) 的1 。急性或短期毒性试验主要用于这一目的, 因为它们给出了一个物种的敏感性的快速迹象。然而, 在它们的自然环境中, 生物体暴露在较长的时间内, 而完整的生命周期, 甚至几代可能会影响到2。此外, 受污染环境中的有机体通常会一次暴露于多个压力源, 这可能相互作用, 可能导致协同效应3。因此, 基于急性单源压力毒性试验计算的安全浓度可能低估了有毒物质在自然环境中的实际风险。因此, 最好还要研究欧洲委员会4、5和环保局 (联合致死) 所倡导的与环境有关的有毒物质的长期和几代效应。国家环境保护局)6,7。特别是在脊椎动物的研究中, 由于脊椎动物的寿命比无脊椎动物模型生物体长, 所以在从事慢性和几代人的暴露研究时, 劳动力、金钱和时间方面的成本很高。因此, 根据研究的问题, 选择最合适的鱼类模型有机体是可取的。此外, 还应提供广泛的脊椎动物种类, 以检验跨物种的反应的普遍性, 以便能够根据最敏感的物种来调整规章。目前, 有必要制定新的、有效的协议, 以短寿命周期为特征的脊椎动物模型物种, 以降低在脊椎动物身上进行慢性和多代暴露的成本7,8。
绿松石鳉鱼Nothobranchius furzeri 是一个有趣的鱼模型, 用于这种长期暴露实验, 因为它的成熟时间短和生命周期 (生成时间少于4周 9).这意味着与其他鱼类模型7相比, 可以在短时间内研究与生态相关的终点, 如成熟时间和繁殖力。此外, 这些鱼生产抗旱的休眠蛋, 在标准条件下储存数年后仍然可行, 从而消除了对连续养殖的需要9。在生态的研究中, 这也意味着复制鱼可以在完全相同的时刻孵化, 导致所有动物的时间同步, 甚至在不同时间生产的鸡蛋批次中。我们建议使用实验室 GRZ 应变进行暴露实验。这种菌株在实验室条件下表现良好, 是纯合的 (除了性染色体), 基因组的特征很好10,11。
在生态研究中, 选择合适的测试浓度范围是很重要的。为此, 可以使用几种互补方法。标称浓度范围可以基于相关物种的灵敏度, 如Nothobranchius guentheri12。或者, 该范围可以基于标准鱼模型的灵敏度, 如斑马鱼 (斑马斑马)2 , 对大多数毒物具有可比性的敏感性 (菲利普et 。(回顾))。结合上述两种选择, 应进行范围发现实验, 选择标称浓度范围。对于急性试验, 研究人员应瞄准100% 的死亡率, 中间死亡率和0% 的死亡后24小时接触毒物的浓度治疗。对于慢性试验, 最好在两周内进行范围发现实验, 以验证在这个参考期内, 测试浓度最高的条件下的幼虫死亡率是否不超过10%。
该议定书可以作为基线, 在furzeri上对水上污染物进行急性和长期接触, 检查压力源在个体和细胞层面的潜在影响。它还可用于执行多源压力研究, 以适应更高的生态相关性, 混合不同的有毒化合物或研究污染与其他自然压力源 (例如捕食) 之间的交互作用或人为压力源 (例如因气候变化而变暖)。

<table>
	<tbody>
		<tr>
			<td>purified water Type 1 (milli Q)</td>
			<td>Millipore</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sea Salt</td>
			<td>Instant Ocean</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>2L plastic tank</td>
			<td>SAVIC</td>
			<td></td>
			<td>Always separate material for control and toxicity treatments</td>
		</tr>
		<tr>
			<td>1L plastic tank (spawning)</td>
			<td>Avamoplast</td>
			<td></td>
			<td>Always separate material for control and toxicity treatments</td>
		</tr>
		<tr>
			<td>nets</td>
			<td>Aqua bilzen</td>
			<td></td>
			<td>Always separate material for control and toxicity treatments</td>
		</tr>
		<tr>
			<td>2L glass jars</td>
			<td>Sepac-Flacover</td>
			<td></td>
			<td>Always separate material for control and toxicity treatments</td>
		</tr>
		<tr>
			<td>0,5L glass jars</td>
			<td>Sepac-Flacover</td>
			<td></td>
			<td>Always separate material for control and toxicity treatments</td>
		</tr>
		<tr>
			<td>Artemia eggs</td>
			<td>Ocean Nutrition</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>chironomus</td>
			<td>Ocean Nutrition</td>
			<td></td>
			<td>frozen</td>
		</tr>
		<tr>
			<td>tricaine</td>
			<td>Sigma aldrich</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>petri dishes</td>
			<td>VWR</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Parafilm</td>
			<td>VWR</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>pipettes</td>
			<td>MLS</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>tweezers</td>
			<td>FST</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>500 &#181;m mesh sieve</td>
			<td>/</td>
			<td></td>
			<td>self-made</td>
		</tr>
		<tr>
			<td>microcentrifuge tube (2ml)</td>
			<td>BRAND</td>
			<td></td>
			<td>To store fish in freezer</td>
		</tr>
		<tr>
			<td>glass vials</td>
			<td>Sigma aldrich</td>
			<td></td>
			<td>For water analysis</td>
		</tr>
		<tr>
			<td>weighing boat</td>
			<td>MLS</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Jiffy 7c pellets</td>
			<td>Jiffy</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>water bath</td>
			<td>Gilac</td>
			<td></td>
			<td>for Ctmax</td>
		</tr>
		<tr>
			<td>liquid nitrogen</td>
			<td>Air liquide</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>digital thermometer</td>
			<td>Testo AG</td>
			<td>testo 926</td>
			<td></td>
		</tr>
		<tr>
			<td>HETO therm heater</td>
			<td>Anker Schmitt</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>calibrated balance</td>
			<td>Mettler-Toledo AG</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>camera</td>
			<td>/</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>platform for camera</td>
			<td>/</td>
			<td></td>
			<td>self-made</td>
		</tr>
		<tr>
			<td>Multiparameter kit</td>
			<td>HACH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Freezer (-80&#176;C)</td>
			<td>Panasonic Ultra low temperature freezer</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Fysio</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>homogenisation buffer</td>
			<td>VWR</td>
			<td></td>
			<td>0.1&#160;M TRIS&#8211;HCl, pH 8.5, 15&#160;% polyvinyl pyrrolidone, 153&#160;&#181;M MgSO4&#160;and 0.2&#160;% Triton X-100</td>
		</tr>
		<tr>
			<td>chloroform:methanol</td>
			<td>Sigma Aldrich</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>glyceryl tripalmitate</td>
			<td>Sigma Aldrich</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>amyloglucosidase</td>
			<td>Sigma Aldrich</td>
			<td>A7420</td>
			<td></td>
		</tr>
		<tr>
			<td>glucose assay reagent</td>
			<td>Sigma Aldrich</td>
			<td>G3293</td>
			<td></td>
		</tr>
		<tr>
			<td>Biorad protein dye</td>
			<td>VWR</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>96-well microtiter plate</td>
			<td>Greiner Bio-one</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>384 microtiter plates</td>
			<td>Greiner Bio-one</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>2 ml glass tubes</td>
			<td>Fiers</td>
			<td></td>
			<td>For fat analysis</td>
		</tr>
		<tr>
			<td>2,5ml eppendorf tubes</td>
			<td>VWR</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>homogeniser</td>
			<td>Ultra-turrax TP 18/10</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>photospectrometer</td>
			<td>Infinite M200 TECAN</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>heater for glass tubes</td>
			<td>Hach COD REACTOR</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>centrifuge</td>
			<td>Eppendorf Centrifuge 5415 R</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Incubator</td>
			<td>Bumako</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

protocol for acute and chronic ecotoxicity testing of the turquoise killifish nothobranchius furzeri

鳉鱼Nothobranchius furzeri 是生态领域新兴的模型有机体, 它在急性和慢性毒性试验中的适用性已经证明。总体而言, 该物种对有毒化合物的敏感性与其他模型物种的敏感程度相同。
这项工作描述了对furzeri的单一和联合应激作用的急性、慢性和几代生物鉴定的协议。由于它的成熟时间短和生命周期, 这脊椎动物模型允许研究终点, 如成熟时间和生育力在四月内。跨代全生命周期暴露试验可以在8月内进行。由于这一物种生产的鸡蛋具有抗旱性, 并且在多年内仍然可行, 所以不需要这种物种的现场养殖, 但在需要时可以招募个人。这些协议旨在测量生命历史特征 (死亡率、生长、繁殖力、体重) 和临界热最大值。

furzeri的严重暴露结果与不同的铜浓度 (如在2.5.2 中计算) 显示 cleardose 响应关系 (图 1)。随着毒物浓度的增加, 死亡率增加。LC50值随着时间的推移而减少, 这意味着随着浓度的降低, 50% 的复制死前会有更多的时间通过。有关furzeri对铜的急性和慢性暴露的详细结果, 以及与其他物种相比, 该物种的灵敏度比较, 我们指的是菲利普et</...

Watch this Scientific Journal Video about Protocol for Acute and Chronic Ecotoxicity Testing of the Turquoise Killifish Nothobranchius furzeri at JoVE.com

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

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

在这项工作中, 我们描述了一种急性, 慢性和几代生物测定, 以研究单一和联合压力源对绿松石鳉鱼Nothobranchiusfurzeri 的影响。本协议旨在研究生命史特征 (死亡、生长、繁殖力、体重) 和临界热最大值。

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

Research

JoVE Journal

Environment

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

Optimized Negative Staining: a High-throughput Protocol for Examining Small and Asymmetric Protein Structure by Electron Microscopy

优化负染：通过电子显微镜检查小和不对称蛋白结构的高通量协议

Structural determination of proteins is rather challenging for proteins with molecular masses between 40 - 200 kDa. Considering that more than half of ...

科研

环境科学

LC-MS Analysis of Human Platelets as a Platform for Studying Mitochondrial Metabolism

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A Protocol for Collecting and Constructing Soil Core Lysimeters

一种用于收集和构建土芯协议测渗计

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Protocol for Microplastics Sampling on the Sea Surface and Sample Analysis

协议塑料微粒采样的海面和样品分析

Microplastic pollution in the marine environment is a scientific topic that has received increasing attention over the last decade. The majority of ...

Ecotoxicological Method with Marine Bacteria Vibrio anguillarum to Evaluate the Acute Toxicity of Environmental Contaminants

Ecotoxicological Method with Marine Bacteria Vibrio anguillarum to Evaluate the Acute Toxicity of Environmental Contaminants

生物毒理学方法与海洋细菌<em&gt;鳗弧菌</em&gt;评估环境污染物的急性毒性

Bacteria are an important component of the ecosystem, and microbial community alterations can have a significant effect on biogeochemical cycling and food ...

Protocol for Assessing the Relative Effects of Environment and Genetics on Antler and Body Growth for a Long-lived Cervid

评估环境和遗传因素对鹿茸和长寿命的鹿的身体生长的相对影响的议定书 》

Cervid phenotype can be placed into one of two categories: efficiency, which promotes survival over extravagant morphometric growth, and luxury, which ...

Dispersion of Nanomaterials in Aqueous Media: Towards Protocol Optimization

纳米材料在水介质中的分散: 对协议优化的研究

The sonication process is commonly used for de-agglomerating and dispersing nanomaterials in aqueous based media, necessary to improve homogeneity and ...

On the Preparation and Testing of Fuel Cell Catalysts Using the Thin Film Rotating Disk Electrode Method

用薄膜旋转圆盘电极法制备和测试燃料电池催化剂的研究

We present a step-by-step tutorial to prepare proton exchange membrane fuel cell (PEMFC) catalysts, consisting of Pt nanoparticles (NPs) supported on a ...

The Hawaii Protocol for Scientific Monitoring of Coffee Berry Borer: a Model for Coffee Agroecosystems Worldwide

夏威夷咖啡莓科学监测协议: 全球咖啡农田模型

Coffee berry borer (CBB) is the most devastating insect pest for coffee crops worldwide. We developed a scientific monitoring protocol that is aimed at ...

Experimental Protocol for Examining Behavioral Response Profiles in Larval Fish: Application to the Neuro-stimulant Caffeine

检测幼虫行为反应剖面的实验协议: 在神经刺激咖啡因中的应用

Fish models and behaviors are increasingly used in the biomedical sciences; however, fish have long been the subject of ecological, physiological and ...