Name: salinity-dependent toxicity assay of silver nanocolloids using medaka eggs
Uploaded: 2016-03-18T13:00:00.000+00:00
Duration: 6 min 29 s
Description: Watch this Scientific Journal Video about Salinity-dependent Toxicity Assay of Silver Nanocolloids Using Medaka Eggs at JoVE.com

We are grateful to Ms. Kaori Shimizu and Mr. Masaki Takasu of the Graduate School of Life Sciences, Toyo University, for their technical support. This project was supported by research grants from the Special Research Foundation and Bio-Nano Electronics Research Centre of Toyo University (to SK); by the Science Research Promotion Fund of the Promotion and Mutual Aid Corporation for Private Schools of Japan (to SK); by the New Project Fund for Risk Assessments, from the Ministry of Economy, Trade and Industry (to SK); by a Grant-in-Aid for Challenging Exploratory Research (award 23651028 to SK); by a Grant-in-Aid for Scientific Research (B) and (C) (award 23310026 and 26340030 to SK); and by a Grant-in-Aid for Strategic Research Base Project for Private Universities (award S1411016 to SK) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

在这项研究中使用的青鳉日本人道分别按照东洋大学的机构准则处理，适当考虑痛苦和不适的缓解。 1.白银纳米胶体（SNC），它们<ol><li>购买纯化SNCs（20毫克/升-1，％纯度99.99，粒 ​​子平均大约悬浮于蒸馏水28.4±8.5纳米直径）。 </li><li>电感耦合等离子体质谱法确认银的纯度和浓度（ICP-MS）根据操作手册8分析。对于ICP-MS的预处理方法分析在第7节描述。 </li></ol> 2. SNC解决方案与不同盐度（银胶和Ag +的混合物）的制备<ol><li>准备60×ERM由60克氯化钠，1.8克氯化钾2.4克氯化钙2·2H 2 O和9.78克硫酸镁4·7H 2 O在1升ULT的rapure水;用1.25％将pH调节至7.0的NaHCO 3在超纯水中。 </li><li>在搅拌25℃的ERM解决方案过夜。 </li><li>混合稀释ERM SNCs。准备40毫升每个SNC-ERM混合溶液。的最终浓度为SNCs的不同浓度的ERM的10毫克/升-1（1倍，5倍，10倍，15倍，20倍，或30倍）。 </li><li>调整的NaHCO 3在超纯水中以SNC-ERM混合溶液至7.0 0.625％的pH值。 pH值的调整是在制备SNC溶液非常重要的，因为银离子释放是通过在酸性条件9促进。 </li><li>使用硝酸银作为SNCs参考化合物。 <ol><li>混合硝酸银用稀释的企业风险管理。在15.7毫克/升的硝酸银浓度制备的40ml的AgNO 3 -ERM混合溶液-1（10毫克/升-1银）在不同浓度的ERM的（1×，5倍，10倍，15倍，20倍，或30倍） 。 注意:要检查银胶毒性， 硝酸银解决方案，这是可溶性银的来源，被用作用于SNCs，这是银胶体和可溶性银的混合物的参考化合物。 </li></ol></li></ol> 3.青鳉文化和鸡蛋收获<ol><li>获取青鳉（O.鳉 ）（橙红色株）（60男性和女性60）。 </li><li>文化作为青鳉组（男20例，女20例为一组）的1X ERM 3升油箱用青鳉流通培养体系。 <ol><li>培养物在下列条件: pH范围培养基:6.2至6.5 光:黑暗周期:16:8小时 培养基的温度:24±0.5℃下 培养基的渗透压:257毫渗</li></ol></li><li> 10:00（每天一次）饲料对卤虫无节幼体青鳉和09:00，11:00，13:00，15:00，17:00养活人工干烧鱼饲料（每日五次）。 <ol><li> A.获取藻万尾。 </li> &lt;李&gt;在塑料烧杯中，准备5升的3.0％的盐溶液。 </li><li> 30克卤虫鸡蛋添加到烧杯中的盐溶液中。 </li><li>孵蛋在25℃48小时用的曝气泵鼓泡（4升/分钟-1）。 </li><li> 48小时后，停止冒泡。 </li><li>使溶液静置5〜10分钟，以阴影线A.分离藻幼体（溶液下部）从未孵化卵和蛋壳（溶液的上部）。 </li><li>除去通过倾析溶液的上层。 </li><li>通过具有283微米开口的筛子过滤溶液的下部，并收集通过对带有198微米的开口的网的无节幼体。 </li><li>饲料无节幼体的青鳉6小时之内。 </li></ol></li><li>女青鳉已经催生后，从女性的身体轻轻取出外部鸡蛋集群或使用SM收集鱼缸底部的蛋人人网（净大小为5厘米×5厘米，孔径0.2毫米×0.2毫米）。 </li><li>冲洗5秒流动自来水蛋群集。 </li><li>所有的冲洗鸡蛋集群添加到30倍ERM解决方案。 </li><li> 1分钟后，除去从溶液中​​簇和放置干燥纸巾和辊轻轻之间的蛋簇。 </li><li>把鸡蛋放回30倍ERM。 </li><li>在解剖显微镜下选择受精卵。 </li><li>地点已由使用镊子选择810鸡蛋1个ERM在六个孔塑料板。 </li><li>在培养箱中直至发育阶段21.孵蛋在25±0.1℃（青鳉胚胎的发育阶段从岩松10的工作所定义。） </li><li>挑出孵化的蛋在发育阶段21在解剖显微镜下。 </li><li>冲洗选鸡蛋1个ERM。 </li><li>若使冲洗鸡蛋暴露实验（第4节）。 </li></ol> 4. SNCs毒性试验或硝酸银</sub&gt;不同盐度ERM <ol><li>冲洗青鳉卵（阶段21）三次测试溶液[SNCs（10毫克/升-1），或硝酸银（15.7毫克/升-1 10毫克/升-1银）在ERM的各浓度（1倍，5倍，10倍，15倍，20倍，或30倍）在pH 7。作为对照，使用1×30×ERM鸡蛋在pH值为7。 </li><li>添加15冲洗鸡蛋各5ml的测试解决方案在六个孔塑料板。 （执行暴露实验三次SNC或使用每种测试溶液硝酸银毒性测试。） </li><li>包裹在铝箔的板。 </li><li>孵育在25℃下在黑暗中包裹板直至孵化或14天。 </li><li>观察暴露的鸡蛋，每24小时的生物变化及死卵（ 图1和2）。 </li><li>交换测试解决方案，每24小时。 </li><li>如下进行观察。 <ol><li>在曝光的第6天，算上心脏率（每15秒）O在解剖显微镜下˚F鳉胚胎通过使用秒表（ 图3a）。 </li><li>在暴露的第6天，通过使用千分尺（ 图3b）测量鳉胚胎解剖显微镜下眼尺寸（直径）。 </li><li>对孵化的一天，通过使用千分尺（ 图3c）测量在解剖显微镜下幼虫的全身长度。 </li><li>算曝光蛋孵化在14天（ 图3d）的总数。 </li></ol></li></ol> 5.从隔离解决方案SNC易溶银，银分析<ol><li>通过在14,000×通过3 kDa的膜过滤器过滤分离从每个SNC溶液（银胶体和可溶性银的混合物）可溶性银 g且4℃，10分钟。使用3 kDa的膜过滤器，以可溶性银从SNCs隔离，因为在1×ERM凝集SNCs的报告平均直径为67.8纳米11第二是银离子的是0.162纳米12; 3 kDa的膜不包括颗粒直径为2nm以上13。 </li><li>根据ICP-MS操作手册8测量在50μl过滤的溶液（=可溶的银浓度）通过ICP-MS分析（ 图3e）的银浓度。对于ICP-MS的预处理方法分析在第7节描述。 </li></ol> 6.银累积的鳉胚胎的测量<ol><li>揭露青鳉鸡蛋（21期），以SNCs或硝酸银作为第4节。 </li><li>在暴露的第6天，通过使用鳉根据在鳉簿 14中描述的协议孵化酶从蛋（ 即 ，dechorion）除去绒毛膜。 </li><li>根据ICP-MS操作手册8（ 图3F）测量由ICP-MS分析dechorioned鸡蛋的银浓度。预处理对于ICP-MS分析法在第7节描述。 </li></ol> 7. ICP-MS分析测量银浓度的<ol><li>添加样本[50微升银溶液（对于银浓度的验证;第1节）; 3 dechorionated胚胎（第5条）;或50微升过滤的溶液（部分5）]到50ml特氟隆烧杯中。 </li><li>加入2.0毫升超纯硝酸50毫升烧杯中。 </li><li>加热上，在110℃的热板中的混合物，直到它变干（约3小时）​​之前。 </li><li>以完全溶解的有机物，加入2.0毫升超纯硝酸和0.5ml过氧化氢的烧杯。 </li><li>在热板上再次加热该混合物，直到它变干（约3小时）​​之前。 </li><li>溶解残留物在4毫升的1.0％的超纯硝酸溶液。 </li><li>传输4毫升溶液到一个离心管中。 </li><li>重复7.6至7.7倍（总共三次）。最终的体积是12.0毫升。 </li><li>测量通过根据操作手册8使用ICP-MS分析的样品（溶于1.0％超纯硝酸）的银浓度。 <ol><li>使用内部和外部的标准溶液（见材料 清单 ）来量化银浓度。内部和外部标准溶液​​是由美国实验室认可协会（A2LA）的认可。银的检出限为0.0018纳克/毫升-1（溶液）和0.016纳克毫克体重-1（胚体）。 </li></ol></li></ol>

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The authors declare that they have no competing financial interests.

青鳉是一种淡水鱼，是非常宽容的海水;它不是众所周知，这种鱼的原有自然栖息地是海水离日本海岸6。因此，青鳉鱼有发达的氯细胞7。这种独特的属性只用鱼的单一品种提供了一种新的方式来测试化学物质的毒性在环境盐度的函数科学家（淡水海水）。 在阶段21获得鳉鸡蛋，鸡蛋必须每天早上收获，并在舞台20.一般选择，青鳉对开始在清晨交配（日出前）?...

自1979年成立了经济合作与发展组织（OECD）的测试准则测试化学品，38测试准则已刊登在准则，对生物系统的影响1第2节。所有测试已经从淡水栖息地，即淡水植物水生生物;藻类;无脊椎动物如水蚤和摇蚊;和鱼类如青鳉，斑马鱼和虹鳟鱼。相比于盐水环境，淡水环境中更直接受到人类经济和工业活动。因此，淡水环境已被优先用于测试，因​​为它们是在距污染风险较高。 在沿海地区，包括河口，盐度苦咸水和海水的条件各不相同，而这些地区往往是由工业活动污染2。沿海地区及其相关湿地的特点是用hIGH生态多样性和生产力。因此，沿海生态系统必须受到保护，免受化学污染。然而，有已经在有限的苦咸水和海水栖息地的生态毒性研究。 Sakaizumi 3就读于日本青鳉鸡蛋甲基汞和盐度之间的毒性相互作用，发现增加供试品溶液的渗透压提高了甲基汞的毒性。 。炭谷等 4用于青鳉鸡蛋探讨垃圾渗滤液的毒性;他们发现，渗滤液对鸡蛋渗透等价的关键是胚胎发育过程中诱导异常。此外，Kashiwada 5日报道，塑料颗粒（直径为39.4纳米）很容易通过微咸水条件下青鳉鸡蛋绒毛膜（15X胚胎饲养介质（ERM））的渗透。 一个典型的小鱼模型，日本青鳉（ 青鳉 </eM&gt;）已在基础生物学和生态毒理学6中。日本青鳉可以住在条件淡水海水，因为他们的高度发达的氯细胞7一应俱全。因此，它们可能是在条件具有广泛的盐度的测试是有用的。

<table><tbody><tr><td>Silver nanocolloids</td><td>Utopia Silver Supplements</td><td></td><td></td></tr><tr><td>NaCl</td><td>Nacalai Tesque, Inc.</td><td>31319-45</td><td>For making ERM</td></tr><tr><td>KCl</td><td>Nacalai Tesque, Inc.</td><td>28513-85</td><td>For making ERM</td></tr><tr><td>CaCl&lt;sub&gt;2&lt;/sub&gt;&amp;middot;2H&lt;sub&gt;2&lt;/sub&gt;O</td><td>Nacalai Tesque, Inc.</td><td>06730-15</td><td>For making ERM</td></tr><tr><td>MgSO&lt;sub&gt;4&lt;/sub&gt;&amp;middot;7H&lt;sub&gt;2&lt;/sub&gt;O</td><td>Nacalai Tesque, Inc.</td><td>21002-85</td><td>For making ERM</td></tr><tr><td>NaHCO&lt;sub&gt;3&lt;/sub&gt;&amp;nbsp;</td><td>Nacalai Tesque, Inc.</td><td>31212-25</td><td>For making ERM</td></tr><tr><td>AgNO&lt;sub&gt;3&lt;/sub&gt;</td><td>Nacalai Tesque, Inc.</td><td>31018-72</td><td></td></tr><tr><td>pH meter</td><td>HORIBA, Ltd.</td><td>F-51S</td><td></td></tr><tr><td>Balance</td><td>Mettler-Toledo International Inc.</td><td>MS204S</td><td></td></tr><tr><td>medaka (&lt;em&gt;Oryzias latipes&lt;/em&gt;) orange-red strain</td><td>National Institute for Environmental Studies</td><td></td><td></td></tr><tr><td>medaka flow-through culturing system</td><td>Meito Suien Co.</td><td>MEITOsystem</td><td></td></tr><tr><td>Artemia salina nauplii eggs</td><td>Japan pet design Co. Ltd</td><td>4975677033759</td><td></td></tr><tr><td>aeration pump</td><td>Japan pet design Co. Ltd</td><td>non-noise w300</td><td></td></tr><tr><td>Otohime larval &amp;beta;-1</td><td>Marubeni Nissin Feed Co. Ltd</td><td>Otohime larval &amp;beta;-1</td><td>Artificial dry fish diet</td></tr><tr><td>dissecting microscope</td><td>Leica microsystems</td><td>M165FC</td><td></td></tr><tr><td>micrometer</td><td>Fujikogaku, Ltd.</td><td>10450023</td><td></td></tr><tr><td>incubator</td><td>Nksystem</td><td>TG-180-5LB</td><td></td></tr><tr><td>shaker</td><td>ELMI Ltd.</td><td>Aizkraukles 21-136</td><td></td></tr><tr><td>6-well plastic plates</td><td>Greiner CELLSTAR</td><td>M8562-100EA</td><td></td></tr><tr><td>aluminum foil</td><td>AS ONE Co.</td><td>6-713-02</td><td></td></tr><tr><td>stopwatch</td><td>DRETEC Co. Ltd.</td><td>SW-111YE</td><td></td></tr><tr><td>3 kDa membrane filter</td><td>EMD Millipore Corporation</td><td>0.5 ml centrifugal-type filter</td><td></td></tr><tr><td>50 ml Teflon beaker</td><td>AS ONE Co.</td><td>33431097</td><td></td></tr><tr><td>Custom claritas standard</td><td>SPEXertificate</td><td>ZSTC-538</td><td>For internal standard</td></tr><tr><td>Custom claritas standard</td><td>SPEXertificate</td><td>ZSTC-622</td><td>For external standard</td></tr><tr><td>ultrapure nitric acid</td><td>Kanto Chemical Co.</td><td>28163-5B</td><td></td></tr><tr><td>hydrogen peroxide&amp;nbsp;</td><td>Kanto Chemical Co.</td><td>18084-1B</td><td>for atomic absorption spectrometry</td></tr><tr><td>ICP-MS</td><td>Thermo Scientific</td><td>Thermo Scientific X Series 2&amp;nbsp;</td><td></td></tr><tr><td>hot plate</td><td>Tiger Co.</td><td>CRC-A300</td><td></td></tr></tbody></table>

salinity-dependent toxicity assay of silver nanocolloids using medaka eggs

盐度是水生环境的一个重要特征。对水生生物有它定义淡水，微咸水，海水的栖息地。化学品和它们的生态风险对水生生物评估的毒性试验在淡水中频繁地进行的，但化学品的水生生物的毒性取决于pH值，温度和盐度。没有方法，然而，用于测试毒性的盐度依赖对水生生物。这里，我们使用青 ​​鳉（ 青鳉 ），因为它们能适应淡水，微咸水和海水。不同浓度的胚胎饲养介质（ERM）（1×，5倍，10倍，15倍，20倍，和30倍）被雇用来测试银nanocolloidal颗粒（SNC），它们的毒性青鳉卵（1×ERM和30倍的ERM有渗透压当量分别淡水和海水）。在六个孔 ​​塑料板，15鳉卵以一式三份在10毫克/升和暴露于SNCs＃8722; 1在pH为7和25°C在黑暗中不同浓度的ERM的。 我们使用解剖显微镜和一个千分尺来测量心脏率达到15秒，眼径6日和孵化天（第4节）幼虫的全身长度。观察胚胎直至孵化或14日;那么，我们每天都计算孵化率为14天（第4节）。要在胚胎银的积累，我们采用电感耦合等离子体质谱测量测试解决方案的银浓度（第5）和dechorionated胚胎（第6）SNCs以青鳉胚胎毒性.The随着盐度明显增加。这种新方法允许我们测试化学物质的毒性在不同的盐度。

盐度对SNC毒性效果非常明显:畸形或死亡的诱导是盐度依赖性的（ 图1和2）。我们在SNC测定表型生物标记（心脏速率，眼睛的大小，完整的体长，和孵化率）（10毫克/升-1）-exposed胚胎。这些表型的生物标志物发现盐度依赖SNC毒性。 心脏率从29.6到32.2次/ 15秒1X各地在控制ERM 30倍不等。然而，他们?...

Watch this Scientific Journal Video about Salinity-dependent Toxicity Assay of Silver Nanocolloids Using Medaka Eggs at JoVE.com

Salinity-dependent Toxicity Assay of Silver Nanocolloids Using Medaka Eggs

Embryonic stages are the most susceptible to xenobiotics. Although chemical toxicity depends on salinity, no method exists to test the salinity dependence of toxicity to aquatic organisms. Here, we describe a new and high-throughput method for determining the salinity dependence of toxicity to aquatic embryos.

白银纳米胶体的盐度依赖性毒力测定使用青鳉鸡蛋

Salinity is an important characteristic of the aquatic environment. For aquatic organisms it defines the habitats of freshwater, brackish water, and ...

salinity-dependent-toxicity-assay-silver-nanocolloids-using-medaka

Universidad San Sebastian

Research

JoVE Journal

Developmental Biology

8.0K 次观看.  Toyo University. Embryonic stages are the most susceptible to xenobiotics. Although chemical toxicity depends on salinity, no method exists to test the salinity dependence of toxicity to aquatic organisms. Here, we describe a new and high-throughput method for determining the salinity dependence of toxicity to aquatic embryos.

视频: Salinity-dependent Toxicity Assay of Silver Nanocolloids Using Medaka Eggs

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

Bacteria are an important component of the ecosystem, and microbial community alterations can have a significant effect on biogeochemical cycling and food webs. Toxicity testing based on microorganisms are widely used because they are relatively quick, reproducible, cheap, and are not associated with ethical issues. Here, we describe an ecotoxicological method to evaluate the biological response of the marine bacterium Vibrio anguillarum. This method assesses the acute toxicity of chemical compounds, including new contaminants such as nanoparticles, as well as environmental samples. The endpoint is the reduction of bacterial culturability (i.e., the capability to replicate and form colonies) due to exposure to a toxicant. This reduction can be generally referred to as mortality. The test allows for the determination of the LC50, the concentration that causes a 50% decrease of bacteria actively replicating and forming colonies, after a 6 h exposure. The culturable bacteria are counted in terms of colony forming units (CFU), and the &#34;mortality&#34; is evaluated and compared to the control. In this work, the toxicity of copper sulphate (CuSO4) was evaluated. A clear dose-response relationship was observed, with a mean LC50 of 1.13 mg/L, after three independent tests. This protocol, compared to existing methods with microorganisms, is applicable in a wider range of salinity and has no limitations for colored/turbid samples. It uses saline solution as the exposure medium, avoiding any possible interferences of growth medium with the investigated contaminants. The LC50 calculation facilitates comparisons with other bioassays commonly applied to ecotoxicological assessments of the marine environment.

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

This work describes a new protocol to assess the ecotoxicity of pollutants, including emerging contaminants such as nanomaterials, using the marine bacterium Vibrio anguillarum. This method allows for the determination of the LC50, or mortality, the concentration that causes a 50% decrease in bacteria culturability, after a 6 h exposure.

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 ...

科研

环境科学

Drug Treatment and In Vivo Imaging of Osteoblast-Osteoclast Interactions in a Medaka Fish Osteoporosis Model

Bone-forming osteoblasts interact with bone-resorbing osteoclasts to coordinate the turnover of bone matrix and to control skeletal homeostasis. Medaka and zebrafish larvae are widely used to analyze the behavior of bone cells during bone formation, degeneration, and repair. Their optical clarity allows the visualization of fluorescently labeled bone cells and fluorescent dyes bound to the mineralized skeletal matrix. Our lab has generated transgenic medaka fish that express the osteoclast-inducing factor Receptor Activator of Nuclear-factor &#954;B Ligand (RANKL) under the control of a heat&#160;shock-inducible promoter. Ectopic expression of RANKL results in the excess formation of activated osteoclasts, which can be visualized in reporter lines with nlGFP expression under the control of the cathepsin K (ctsk) promoter. RANKL induction and ectopic osteoclast formation leads to severe osteoporosis-like phenotypes. Compound transgenic medaka lines that express ctsk:nlGFP in osteoclasts, as well as mCherry under the control of the osterix (osx) promoter in premature osteoblasts, can be used to study the interaction of both cell types. This facilitates the in vivo observation of cellular behavior under conditions of bone degeneration and repair. Here, we describe the use of this system to test a drug commonly used in human osteoporosis therapy and describe a protocol for live imaging. The medaka model complements studies in cell culture and mice, and offers a novel system for the in vivo analysis of drug action in the skeletal system.

Drug Treatment and In Vivo Imaging of Osteoblast-Osteoclast Interactions in a Medaka Fish Osteoporosis Model

Small laboratory fish have become popular models for bone research on the mechanisms underlying human bone disorders and for the screening of bone-modulating drugs. In this report, we describe a protocol to assess the effect of alendronate on bone cells in medaka larvae with osteoporotic lesions.

戒毒治疗和<em&gt;在体内</em&gt;在青鳉鱼骨质疏松模型成骨破骨细胞相互作用的成像

Bone-forming osteoblasts interact with bone-resorbing osteoclasts to coordinate the turnover of bone matrix and to control skeletal homeostasis. Medaka ...

发育生物学

Use of Autometallography to Localize and Semi-Quantify Silver in Cetacean Tissues

Silver nanoparticles (AgNPs) have been extensively used in commercial products, including textiles, cosmetics, and health care items, due to their strong antimicrobial effects. They also may be released into the environment and accumulate in the ocean. Therefore, AgNPs are the major source of Ag contamination, and public awareness of the environmental toxicity of Ag is increasing. Previous studies have demonstrated the bioaccumulation (in producers) and magnification (in consumers/predators) of Ag. Cetaceans, as the apex predators of ocean, may have been negatively affected by the Ag/Ag compounds. Although the concentrations of Ag/Ag compounds in cetacean tissues can be measured by inductively coupled plasma mass spectroscopy (ICP-MS), the use of ICP-MS is limited by its high capital cost and the requirement for tissue storage/preparation. Therefore, an autometallography (AMG) method with an image quantitative analysis by using formalin-fixed, paraffin-embedded (FFPE) tissue may be an adjuvant method to localize Ag distribution at the suborgan level and estimate the Ag concentration in cetacean tissues. The AMG positive signals are mainly brown to black granules of various sizes in the cytoplasm of proximal renal tubular epithelium, hepatocytes, and Kupffer cells. Occasionally, some amorphous golden yellow to brown AMG positive signals are noted in the lumen and basement membrane of some proximal renal tubules. The assay for estimating the Ag concentration is named the Cetacean Histological Ag Assay (CHAA), which is a regression model established by the data from image quantitative analysis of the AMG method and ICP-MS. The use of AMG with CHAA to localize and semi-quantify heavy metals provides a convenient methodology for spatio-temporal and cross-species studies.

A protocol is presented to localize Ag in cetacean liver and kidney tissues by autometallography. Furthermore, a new assay, named the cetacean histological Ag assay (CHAA) is developed to estimate the Ag concentrations in those tissues.

Autometallography 在鲸目组织中的定位和半定量银的应用

Silver nanoparticles (AgNPs) have been extensively used in commercial products, including textiles, cosmetics, and health care items, due to their strong ...

A Small-Scale Setup for Algal Toxicity Testing of Nanomaterials and Other Difficult Substances

Ecotoxicity data is a requirement for pre- and post-market registration of chemicals by European and international regulations (e.g., REACH). The algal toxicity test is frequently used in regulatory risk assessment of chemicals. In order to achieve high reliability and reproducibility the development of standardized guidelines is vital. For algal toxicity testing, the guidelines require stable and uniform conditions of parameters such as pH, temperature, carbon dioxide levels and light intensity. Nanomaterials and other so-called difficult substances can interfere with light causing a large variation in results obtained hampering their regulatory acceptance. To address these challenges, we have developed LEVITATT (LED Vertical Illumination Table for Algal Toxicity Tests). The setup utilizes LED illumination from below allowing for a homogenous light distribution and temperature control while also minimizing intra-sample shading. The setup optimizes the sample volume for biomass quantification and does at the same time ensure a sufficient influx of CO2 to support exponential growth of the algae. Additionally, the material of the test containers can be tailored to minimize adsorption and volatilization. When testing colored substances or particle suspensions, the use of LED lights also allows for increasing the light intensity without additional heat generation. The compact design and minimal equipment requirements increase the possibilities for implementation of the LEVITATT in a wide range of laboratories. While compliant with standardized ISO and OECD guidelines for algal toxicity testing, LEVITATT also showed a lower inter-sample variability for two reference substances (3,5-Dicholorophenol and K2Cr2O7) and three nanomaterials (ZnO, CeO2, and BaSO4) compared to Erlenmeyer flasks and microtiter plates.

We demonstrate algal toxicity testing for difficult substances (e.g., colored substances or nanomaterials) using a setup illuminated vertically with an LED.

纳米材料和其他难物质藻类毒性试验的小型装置

Ecotoxicity data is a requirement for pre- and post-market registration of chemicals by European and international regulations (e.g., REACH). The algal ...

Lethality Bioassay Using Artemia salina L.

Natural products have been used since ancient times to produce medicines. Nowadays, there are plenty of chemotherapeutic drugs obtained from natural sources and used against a plethora of diseases. Unfortunately, most of these compounds often display systemic toxicity and adverse effects. In order to better evaluate the tolerability of selected potentially bioactive samples, brine shrimp (Artemia salina) is generally used as a model in lethality studies. The A. salina test is based on the ability of the studied bioactive compounds to kill the microcrustaceans in their larval stage (nauplii). This method represents a convenient starting point for cytotoxicity studies, as well as for the general toxicity screening of synthetic, semisynthetic, and natural products. It can be considered a simple, quick, and low-cost assay, compared to many other assays (in vitro cells or yeast strains, zebrafish, rodents) generally suitable for the aforementioned purposes; moreover, it can be easily performed even without any specific training. Overall, A. salina assay represents a useful tool for the preliminary toxicity evaluation of selected compounds and the bio-guided fractionation of natural product extracts.

Lethality Bioassay Using Artemia salina L.

This work aims to evaluate and review the Artemia salina lethality bioassay procedure, also identified as brine shrimp lethality assay. This simple and cheap method gives information about the general toxicity (considered as a preliminary toxicity evaluation) of samples, namely, natural products.

使用 卤虫 盐L的致死性生物测定。

Natural products have been used since ancient times to produce medicines. Nowadays, there are plenty of chemotherapeutic drugs obtained from natural ...

化学

Long-term Lethal Toxicity Test with the Crustacean Artemia franciscana

Our research activities target the use of biological methods for the evaluation of environmental quality, with particular reference to saltwater/brackish water and sediment. The choice of biological indicators must be based on reliable scientific knowledge and, possibly, on the availability of standardized procedures. In this article, we present a standardized protocol that used the marine crustacean Artemia to evaluate the toxicity of chemicals and/or of marine environmental matrices. Scientists propose that the brine shrimp (Artemia) is a suitable candidate for the development of a standard bioassay for worldwide utilization. A number of papers have been published on the toxic effects of various chemicals and toxicants on brine shrimp (Artemia). The major advantage of this crustacean for toxicity studies is the overall availability of the dry cysts; these can be immediately used in testing and difficult cultivation is not demanded1,2. Cyst-based toxicity assays are cheap, continuously available, simple and reliable and are thus an important answer to routine needs of toxicity screening, for industrial monitoring requirements or for regulatory purposes3. The proposed method involves the mortality as an endpoint. The numbers of survivors were counted and percentage of deaths were calculated. Larvae were considered dead if they did not exhibit any internal or external movement during several seconds of observation4. This procedure was standardized testing a reference substance (Sodium Dodecyl Sulfate); some results are reported in this work. This article accompanies a video that describes the performance of procedural toxicity testing, showing all the steps related to the protocol.

Long-term Lethal Toxicity Test with the Crustacean Artemia franciscana

This study concerns the development and standardization of a valuable methodological protocol to determine long-term (14 days) lethal toxicity exerted by chemical substances, industrial wastewater or sewage and liquid environmental samples on the saltwater crustacean, Artemia franciscana.

致命的甲壳类动物长期毒性试验卤虫franciscana</em

Our research activities target the use of biological methods for the evaluation of environmental quality, with particular reference to saltwater/brackish ...

生物学

Rapid Evaluation of Toxicity of Chemical Compounds Using Zebrafish Embryos

The zebrafish is a widely used vertebrate model organism for the disease and phenotype-based drug discovery. The zebrafish generates many offspring, has transparent embryos and rapid external development. Zebrafish embryos can, therefore, also be used for the rapid evaluation of toxicity of the drugs that are precious and available in small quantities. In the present article, a method for the efficient screening of the toxicity of chemical compounds using 1-5-day post fertilization embryos is described. The embryos are monitored by stereomicroscope to investigate the phenotypic defects caused by the exposure to different concentrations of compounds. Half-maximal lethal concentrations (LC50) of the compounds are also determined. The present study required 3-6 mg of an inhibitor compound, and the whole experiment takes about 8-10 h to be completed by an individual in a laboratory having basic facilities. The current protocol is suitable for testing any compound to identify intolerable toxic or off-target effects of the compound in the early phase of drug discovery and to detect subtle toxic effects that may be missed in the cell culture or other animal models. The method reduces procedural delays and costs of drug development.

Zebrafish embryos are used for evaluating the toxicity of chemical compounds. They develop externally and are sensitive to chemicals, allowing detection of subtle phenotypic changes. The experiment only requires a small amount of compound, which is directly added to the plate containing embryos, making the testing system efficient and cost-effective.

利用斑马鱼胚胎对化合物毒性的快速评价

The zebrafish is a widely used vertebrate model organism for the disease and phenotype-based drug discovery. The zebrafish generates many offspring, has ...

医学

行为的：评估成年斑马鱼神经毒性作用的测试电池

Neurotoxicity Assessment in Adult Danio rerio using a Battery of Behavioral Tests in a Single Tank

The presence of neuropathological effects proved to be, for many years, the main endpoint for assessing the neurotoxicity of a chemical substance. However, in the last 50 years, the effects of chemicals on the behavior of model species have been actively investigated. Progressively, behavioral endpoints were incorporated into neurotoxicological screening protocols, and these functional outcomes are now routinely used to identify and determine the potential neurotoxicity of chemicals. Behavioral assays in adult zebrafish provide a standardized and reliable means to study a wide range of behaviors, including anxiety, social interaction, learning, memory, and addiction. Behavioral assays in adult zebrafish typically involve placing the fish in an experimental arena and recording and analyzing their behavior using video tracking software. Fish can be exposed to various stimuli, and their behavior can be quantified using a variety of metrics. The novel tank test is one of the most accepted and widely used tests to study anxiety-like behavior in fish. The shoaling and social preference tests are useful in studying the social behavior of zebrafish. This assay is particularly interesting since the behavior of the entire shoal is studied. These assays have proven to be highly reproducible and sensitive to pharmacological and genetic manipulations, making them valuable tools for studying the neural circuits and molecular mechanisms underlying behavior. Additionally, these assays can be used in drug screening to identify compounds that may be potential modulators of behavior.
We will show in this work how to apply behavioral tools in fish neurotoxicology, analyzing the effect of methamphetamine, a recreational drug, and glyphosate, an environmental pollutant. The results demonstrate the significant contribution of behavioral assays in adult zebrafish to the understanding of the neurotoxicological effects of environmental pollutants and drugs, in addition to providing insights into the molecular mechanisms that may alter neuronal function.

作者聚焦：成年斑马鱼环境和药物诱导的行为变化评估 

Here, we present a comprehensive behavioral test battery, including the novel tank, Shoaling, and social preference tests, to effectively determine the potential neurotoxic effects of chemicals (e.g., methamphetamine and glyphosate) on adult zebrafish using a single tank. This method is relevant to neurotoxicity and environmental research.

白银纳米胶体的盐度依赖性毒力测定使用青鳉鸡蛋

摘要

探索更多视频

此视频中的章节

生物毒理学方法与海洋细菌

戒毒治疗和

Autometallography 在鲸目组织中的定位和半定量银的应用

纳米材料和其他难物质藻类毒性试验的小型装置

使用卤虫盐L的致死性生物测定。

致命的甲壳类动物长期毒性试验卤虫franciscana

利用斑马鱼胚胎对化合物毒性的快速评价

在单个水箱中使用一系列行为测试对成年 Danio rerio 进行神经毒性评估

在单个水箱中使用一系列行为测试对成年 Danio rerio 进行神经毒性评估

在单个水箱中使用一系列行为测试对成年 Danio rerio 进行神经毒性评估

白银纳米胶体的盐度依赖性毒力测定使用青鳉鸡蛋

摘要

探索更多视频

此视频中的章节

生物毒理学方法与海洋细菌

戒毒治疗和

Autometallography 在鲸目组织中的定位和半定量银的应用

纳米材料和其他难物质藻类毒性试验的小型装置

使用 卤虫 盐L的致死性生物测定。

致命的甲壳类动物长期毒性试验卤虫franciscana

利用斑马鱼胚胎对化合物毒性的快速评价

在单个水箱中使用一系列行为测试对成年 Danio rerio 进行神经毒性评估

在单个水箱中使用一系列行为测试对成年 Danio rerio 进行神经毒性评估

在单个水箱中使用一系列行为测试对成年 Danio rerio 进行神经毒性评估

使用卤虫盐L的致死性生物测定。