Name: basic methods for the study of reproductive ecology of fish in aquaria
Uploaded: 2017-07-20T13:00:00.000+00:00
Duration: 7 min 25 s
Description: Watch this Scientific Journal Video about Basic Methods for the Study of Reproductive Ecology of Fish in Aquaria at JoVE.com

感谢S.Yokoyama协助收集鱼。我们也非常感谢瓦川村有关饲养方法的有用建议。本研究由日本科学促进会（KAKENHI）通过授予TS（24370006和16K07507）的资助

收集和运送活鱼注意:本协议描述了如何从具有深度≥15米的气囊收集具有气囊的鱼。快速输送到表面会导致气囊膨胀，压力变化会严重危害或杀死鱼类。谨慎是必要的，因为在第一步中对鱼类造成的损害将对其生存和后期行为产生不利影响。 <ol><li>在与SCUBA潜水之前，请收集以下材料:适合捕获水下目标物种的手网;双重聚乙烯袋，足够大的鱼;橡胶带（ø80 mm x 6 mm）;用于在表面充气收集袋的氧气瓶;作为抗微生物剂的Elbagin包含10％的硝基苯乙烯钠;聚苯乙烯泡沫箱，其中放置收集袋;移液管;并且如有必要（参见步骤1.4），长度为的绳子相当于最大潜水深度，以及至少2公斤的重量。 </li><li>潜水时用手网收集目标鱼类;同时仍然在水下，将捕获的鱼放在聚乙烯袋中，并用橡皮筋绑扎袋子的口。如果将鱼运送到水族馆需要≥1天的时间，还可以从其他袋子中的自然栖息地中保存水，以用于更新养鱼用水。 注意:为了安全起见，水下任务必须由至少两个人进行。 </li><li>以不超过1米/分钟的速度从收集点收集鱼的表面:在10米或更深的深度处，每2米停止上升1 - 5分钟;并从10米的深度到每1米的表面停止1 - 5分钟。当捕获的鱼不能保持其在袋中的浮力（ 即 ，如果它们在膨胀的气体膀胱浮起来的同时仍然试图向底部游泳时），或者保持相同的深度1-5分钟或de更深一点到2米。一旦鱼似乎恢复了浮力，恢复上升到地面。 注意:如果SCUBA罐中的空气有可能在上升时耗尽，则将绳索固定在聚乙烯袋上，并附加≥2kg的重量;在水下操作者安全上升之后，以与上述相同的速度将其拉到表面。 </li><li>在海滩或船上，表面处理后，每个聚乙烯袋的水中溶解10 ppm的Elbagin。 注意:虽然一些鱼可能在被带到地面之后可能无法保持其浮力，但大多数鱼可能会在一天之内恢复。 </li><li>如果收集袋中的鱼的密度很高，则可以在多袋之间划分鱼，以防止在运输过程中彼此相互碰撞而彼此相互损害。如果鱼是侵略性物种，请将它们分开包裹。 </li><li>使用氧气充气收集袋，并再次使用r密封袋的口ubberbands。当将氧气引入袋中时，将喷嘴降低到袋子的水中以增加溶解氧含量;完全膨胀的袋子应该是1/4满水。 </li><li>将收集袋放在聚苯乙烯泡沫箱中，以保持水温稳定，并在黑暗条件下降低鱼的压力。 </li><li>如果将鱼运送到水族馆需要≥1天的时间，每天一次可以在每个养鱼袋中换入1/4至1/3的水分，使用从栖息地保留的水加入10 ppm Elbagin ，并用氧气重新包装。每天用手网或移液管清除底部的死鱼和排泄物。 注意:如果涉及飞机运输，在运输之前等待至少1天将鱼适应到表面的空气压力，因为在短期内两步压力变化（从水下到地面，表面到上部空气）可能会产生负面影响鱼的生存。 </l<html>我&gt; </ol>将鱼适应水族馆<ol><li>将含有鱼的袋子在水族箱中漂浮30分钟以平衡水温。 </li><li>在10分钟的时间内，用水族箱水逐渐更换袋子水，以避免水分化学（ 例如 ，pH，盐度）突然变化引起的冲击。 注意:由于水分化学差异太大而引起震惊的鱼可能会显示身体颜色和/或行为的异常变化。适应期间仔细监测。 </li><li>将10 ppm Elbagin溶解在水族馆水中。 </li><li>此后，每天一次，持续3天，更新1/3的水族馆水，将10ppm的Elbagin添加到更换的水中。 </li><li>最后，通过一天一次交换一半的水族馆水，直到颜色消失，开始消除Elbagin。 </li></ol> 3.注射可见植入弹性体（VIE）标签以识别个体鱼T"&gt;注:在这项工作中，个别鱼使用VIE标签标识;对于示例，请参阅弗雷德里克·10，奥尔森和Vøllestad11，和勒布朗和诺克斯12此外，如果研究的物种是足够大的手握住，步骤3.2中使用的手术台将不是必需的。 <ol><li>确定每个人的注射位置和标签颜色。最安全的选择是在身体的厚背部或尾部部位选择一个注射点，并避免将其注射到可能刺穿内部器官的腹部。 注意: 图1中描述了编号系统:如果研究物种足够大以注入8个可能的位置，则该系统将允许使用单一颜色识别154个个体。 VIE标签的十种颜​​色可商购。根据鱼的身体颜色选择可区分的颜色。 </li><li> &lt;强&gt;对于小于可以握在手中的鱼，准备如下的外科手术台（也参见Kinkel 等人 13 ）（ 图2 ）。 <ol><li>切出5厘米长×5厘米宽的软海绵，比培养皿的高度至少低5-10毫米。 </li><li>在大约5 - 10 mm深度的海绵上切割一个凹槽，并将其宽度调整为鱼的近似身体宽度。将聚氯乙烯（PVC）板（0.3mm厚）切割成5cm×1.5cm宽，并将其弯曲成谷形折叠（或M形）。 </li><li>将开槽的PVC板放在开槽的海绵上，然后将海绵放入培养皿（ø160 mm，30 mm深）。使用回收罐的水填充培养皿，直到PVC板充分浸泡。 </li></ol></li><li>根据VIE标签手册准备VIE标签。混合弹性体材料，并将混合物加入到3-mL syri中配备29号针头，包含在套件中。 </li><li>在执行标签注射时准备使用两个水箱:一个用于麻醉，一个用于恢复;调整这些的温度和盐度浓度以匹配饲养水族馆的水。使用带有空气石头的空气泵将回水罐中的水温和循环。 </li><li>通过将2-甲基喹啉与等体积的99.5％乙醇混合来制备麻醉液;将其加入麻醉罐中以达到18ppm的浓度。 注意:麻醉液的最佳浓度将取决于个体的物种和体型。因此，提前检查研究物种的最佳浓度。 </li><li>将各个鱼转移到麻醉池，让它们变得麻醉:就是说，当鱼缸外面被点击时，等待鱼不会被触动或水的振动反应。当身体颜色在麻醉时变化在许多鱼中，也仔细监测身体颜色，并判断鱼是否从其变化中麻醉。因为鱼有可能跳出麻醉罐，使用透明的亚克力板覆盖，至少直到鱼开始缓慢移动。 注意:根据研究种类及其体型，微调麻醉和/或调整麻醉液浓度所需的时间。如果手术运动被麻醉所阻止，那么鱼会死亡的风险很高。 </li><li>如果身体变暖，鱼就会变弱，麻醉鱼时，手指会保持在比较冷的水中。 </li><li>从麻醉坦克提起鱼，并快速测量或记录必要的数据（总长度，性别等 ）。如果鱼在测量过程中似乎恢复，请立即将其转移回麻醉池。 </li><li>将鱼转移到手术台。将鱼腹侧放在t中他的PVC槽。如果体型非常小，则在注射时使用双目显微镜。如果研究种类足够大，可以握在手中，则在被保持时注入VIE标签。 </li><li>将针的斜面朝向外部并指向鱼头。皮下注射针，或多或少地平行于身体，并尽可能靠近皮肤下方。根据鱼体尺寸和最终看到标签的容易程度调整插入深度。 </li><li>在取针时注入VIE标签，并在针斜度到达针入口点之前停止注射（如果在相对较宽的区域注入标签，将很容易辨别）。 </li><li>完成标签注射后，立即将鱼转移到回收罐。如果恢复看起来太慢，用手轻轻地循环水。 </li><li>恢复后，将鱼返回饲养水族馆，并继续添加10 ppm Elbagin到水中3天。 注意:在VIE标签的低可见度条件下，UVA滤光灯将有助于识别标签。 </li></ol> 4.计数脱胶胶鸡蛋<ol><li>为了创建一个人造卵巢，鱼可以将粘合蛋沉积，将不透明的PVC管（ø5厘米，6厘米长）切成两半，垂直于直径。 注意:上述管道尺寸适用于产卵相对较小的虫虫T. marinae 。因此，调整适合研究种类的PVC管尺寸和形状。 </li><li>在防水片上打印5 mm x 5 mm格栅，并将其切割成适合PVC管道的内部区域。 </li><li>使用橡皮筋将防水片固定在PVC管的内部。 注意:如果女性难以在防水片上附上鸡蛋，并且鸡蛋从纸张上落下，则使纸张用砂纸进行纹理化。 </li><li>用一层沙子覆盖水族馆底部，1- 厚2厘米。将大约三分之一的PVC管倾斜地插入砂中，将板材固定侧向下。 </li><li> 鱼成功产卵后，从PVC管中取出蛋块，将其放入带有水族馆水的培养皿中。将一张新的纸张固定在PVC管内，并将其放回水族箱的底部。最后，通过拍摄鸡蛋块来计数鸡蛋。 <ol><li>或者，如果在产卵后进行亲子蛋保育的观察，小心不要将鸡蛋从水中取出。相反，使用培养皿与水族馆水一起舀起鸡蛋和床单，快速拍摄鸡蛋质量，然后仔细地将鸡蛋块重新吸收到同一个PVC管中，然后将其全部恢复到原来的位置水族箱底部。最后，使用图片计数鸡蛋。 </li></ol></li><li> 使用ImageJ计数鸡蛋（ 图3 </stro纳克&gt;）。这些自动和手动细胞计数方法在以下详细说明:基本细胞计数（芝加哥大学综合光显微核心设施http://digital.bsd.uchicago.edu/image_j.php）;粒子分析（http://imagej.net/Particle_Analysis）;单元格计数器（https://imagej.net/Cell_Counter）。 <ol><li>否则，如果鸡蛋密集沉积，则通过计算面积密度比估计蛋数:打印5×5 mm 2网格中的鸡蛋计算鸡蛋密度，用ImageJ测量鸡蛋覆盖的表面积，并估算每单位和表面积以上密度的鸡蛋总数。以下解释了ImageJ的上述表面积测量方法:Set scale ...（Research Services Branch，https://imagej.nih.gov/ij/docs/menus/analyze.html）;它解释了如何定义图像的空间尺度;使用ImageJ测量表面积（Keene州立大学，学术技术，https://dept.keene.edu/at/2011/04/15/by-matthew-ragan/）。 注意:如果鸡蛋是透明的，黑色防水片更适合自动计数方法，因为ImageJ可以单独识别鸡蛋的轮廓。 </li></ol></li></ol>

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I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behaviorally+induced+maternal+stress+in+a+fish+influences+progeny+quality+by+a+hormonal+mechanism.">Behaviorally induced maternal stress in a fish influences progeny quality by a hormonal mechanism.</a> Ecology. 79 (6), 1873-1883 (1998).</li><li>Knaepkens, G., Bruyndoncx, L., Coeck, J., Eens, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spawning+habitat+enhancement+in+the+European+bullhead+(Cottus+gobio),+an+endangered+freshwater+fish+in+degraded+lowland+rivers.">Spawning habitat enhancement in the European bullhead (Cottus gobio), an endangered freshwater fish in degraded lowland rivers.</a> Biodivers Conserv. 13 (13), 2443-2452 (2004).</li><li>Shiogaki, M., Dotsu, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+life+history+of+the+gobiid+fish,+Zonogobius+boreus.">The life history of the gobiid fish, Zonogobius boreus.</a> Bulletin of the Faculty of Fisheries, Nagasaki University. 37, 1-8 (1974).</li><li>Meunier, B., Yavno, S., Ahmed, S., Corkum, L. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=First+documentation+of+spawning+and+nest+guarding+in+the+laboratory+by+the+invasive+fish,+the+round+goby+(Neogobius+melanostomus).">First documentation of spawning and nest guarding in the laboratory by the invasive fish, the round goby (Neogobius melanostomus).</a> J Great Lakes Res. 35 (4), 608-612 (2009).</li></ol>

作者没有什么可以披露的。

许多鱼类的生殖生态常常通过实验饲养来揭示。特别是，变性6，8，14，择偶15,16和种内竞争7,17一直使用水族馆保持鱼类详细调查的频繁的话题。此外，在水族箱观察到一些结果已经在后面的字段8，18</sup...

鱼类的形态学，生态学和行为学表​​现出明显的自适应进化1 。特别是与生殖相关的生态特征特别多样，其中大部分可以直接受个人健身的影响2 。为了深入了解导致不同鱼类独特特征演变的选择性压力，使用活鱼直接观察生殖和社会行为通常有助于证实理论假设。 然而，对鱼类的连续现场观察可能需要专门的难以维护的水下设备和设施。在这些情况下，野生捕捞的水族馆饲养的鱼的观察是有帮助3，4，5。此外，鱼类行为的有效观察，其他罕见或困难ULT在自然条件下，观察可以通过在水族箱6，7，8操纵实验成为可能。通过最大程度地减少人为压力和身体伤害，在良好条件下养鱼是准确生态调查的关键。 侏儒虾青y素总长 23-25毫米，分布在西太平洋，安静，庇护的海湾，深度9-26米9 。在这项工作中， T. marinae被用作一个模型，用于描述使用独立的水下呼吸器（SCUBA），鱼类运输和最终驯化鱼类进行直接观察的鱼类收集系列的基本技术的研究物种的生殖行为和生态学。

<table><tbody><tr><td>Hand net</td><td>Nisso</td><td>AQ-17</td><td>Select for the target species size.</td></tr><tr><td>Polyethylene bag</td><td>San-U Fish Farm</td><td>8194</td><td></td></tr><tr><td>Rubber band</td><td>ESCO Co. LTD.</td><td>78-0420-64</td><td>&amp;oslash; 80 mm x 6 mm</td></tr><tr><td>Oxygen cylinder</td><td>N/A</td><td>N/A</td><td>Oxgen cylinder for diving equipment suits.</td></tr><tr><td>Elbagin</td><td>Japan Pet Design Co. Ltd.</td><td>75950</td><td>Pafurazine F (provided from same company) is equivalent drug to Elbagin.</td></tr><tr><td>Polystyrene foam box</td><td>N/A</td><td>N/A</td><td></td></tr><tr><td>Pipette</td><td>AS ONE</td><td>1-8625-04</td><td></td></tr><tr><td>Rope</td><td>Mizukami Kinzoku Co. LTD.</td><td>95301601</td><td></td></tr><tr><td>Weight</td><td>N/A</td><td>N/A</td><td>Weight for diving equipment suits.</td></tr><tr><td>Water tank</td><td>N/A</td><td>N/A</td><td></td></tr><tr><td>Air pump</td><td>KOTOBUKI</td><td>4972814 062115</td><td></td></tr><tr><td>Air stone</td><td>KOTOBUKI</td><td>4972814 232204</td><td></td></tr><tr><td>2-Methylquinoline&amp;nbsp;</td><td>Wako</td><td>170-00376</td><td></td></tr><tr><td>Ethanol (99.5)</td><td>Wako</td><td>057-00456</td><td></td></tr><tr><td>Visible implant elastomer tag kit</td><td>Northwest Marine Technology</td><td>N/A</td><td>http://www.nmt.us/products/vie/vie.shtml</td></tr><tr><td>Soft sponge</td><td>N/A</td><td>N/A</td><td></td></tr><tr><td>PVC board</td><td>N/A</td><td>N/A</td><td>0.3 mm thickness is easy to use.</td></tr><tr><td>Petri dish</td><td>N/A</td><td>N/A</td><td>A large one, such as&amp;nbsp; &amp;oslash; 160 mm and 30 mm depth, is convenient for the injection of the VIE tag.</td></tr><tr><td>Transparent acrylic board</td><td>N/A</td><td>N/A</td><td></td></tr><tr><td>UVA filtered light&amp;nbsp;</td><td>N/A</td><td>N/A</td><td></td></tr><tr><td>PVC pipe</td><td>N/A</td><td>N/A</td><td>&amp;oslash; 5 cm</td></tr><tr><td>Waterproof sheet</td><td>SOMAR Corp.</td><td>3EKW03</td><td>The film for the plain copier.</td></tr><tr><td>Sand</td><td>N/A</td><td>N/A</td><td></td></tr><tr><td>Stereo microscope</td><td>N/A</td><td>N/A</td><td></td></tr><tr><td>Camera</td><td>N/A</td><td>N/A</td><td></td></tr></tbody></table>

basic methods for the study of reproductive ecology of fish in aquaria

当持续的实地调查是不可能的时候，养殖观察对于揭示鱼行为和生态学方面是有价值的。在这里，描述了一系列基本技术，以便观察野生捕获的小型鱼的生殖行为，作为模型，保存在水族箱中。该方法着重于三个步骤:收集，运输和观察底物spavner的生殖生态学。活鱼收集和运输的基本方面是（1）防止对鱼的伤害，（2）仔细适应水族馆。收集活鱼时，预防伤害如划痕或突然变化的水压是必不可少的，因为任何物理伤害可能会对鱼的生存和后期行为产生负面影响。仔细适应水族箱可降低发病死亡率，减轻运输冲击。饲养期间的观察包括（1）识别个人双鱼和（2）监测产卵，对鱼或鸡蛋没有负面影响，从而对研究种的生殖生态进行详细的调查。皮下注射可见植入弹性体（VIE）标签是用于随后鉴定个体鱼的精确方法，并且可以与大范围的鱼一起使用，对其生存和行为影响最小。如果研究物种是沉积粘蛋的底物，那么由聚氯乙烯（PVC）管道构建的人造巢场，加上可拆卸的防水片将有助于计数和监测鸡蛋，减少调查人员对巢穴的影响和鱼的保卫行为。虽然这种基本方法需要在研究文章中很少提及的技术，但它们是进行需要养殖野生鱼类的实验的基础。

按照上述方法，分别于2014年4月，2015年和2016年在日本鹿儿岛县Am美大岛境外收集了41，15和96个海棠的个体（ 表1 ）。在每种情况下，25（61％），14（93％）和91（95％）个人住在直到在水族馆里存放鸡蛋。福田等人报道3 ，在2014年观察期结束前只有一只鱼死亡，鱼类产卵否则在捕获后7天开始出现，表明个体在良好条件下饲?...

Watch this Scientific Journal Video about Basic Methods for the Study of Reproductive Ecology of Fish in Aquaria at JoVE.com

Basic Methods for the Study of Reproductive Ecology of Fish in Aquaria

描述了一系列基本方法来研究养殖在水族箱中的鱼类的生殖生态。这些是使用SCUBA收集鱼类，运送活鱼以及观察保持在水族箱中的野生捕捞鱼的繁殖行为的有用方案。

鱼类生殖生态学研究的基本方法

Captive-rearing observations are valuable for revealing aspects of fish behavior and ecology when continuous field investigations are impossible. Here, a ...

basic-methods-for-the-study-of-reproductive-ecology-of-fish-in-aquaria

Research

JoVE Journal

Behavior

11.5K 次观看.  Tokyo University of Marine Science and Technology. 描述了一系列基本方法来研究养殖在水族箱中的鱼类的生殖生态。这些是使用SCUBA收集鱼类，运送活鱼以及观察保持在水族箱中的野生捕捞鱼的繁殖行为的有用方案。 

视频: Basic Methods for the Study of Reproductive Ecology of Fish in Aquaria

Development of New Methods for Quantifying Fish Density Using Underwater Stereo-video Tools

The use of video camera systems in ecological studies of fish continues to gain traction as a viable, non-extractive method of measuring fish lengths and estimating fish abundance. We developed and implemented a rotating stereo-video camera tool that covers a full 360 degrees of sampling, which maximizes sampling effort compared to stationary camera tools. A variety of studies have detailed the ability of static, stereo-camera systems to obtain highly accurate and precise measurements of fish; the focus here was on the development of methodological approaches to quantify fish density using rotating camera systems. The first approach was to develop a modification of the metric MaxN, which typically is a conservative count of the minimum number of fish observed on a given camera survey. We redefine MaxN to be the maximum number of fish observed in any given rotation of the camera system. When precautions are taken to avoid double counting, this method for MaxN may more accurately reflect true abundance than that obtained from a fixed camera. Secondly, because stereo-video allows fish to be mapped in three-dimensional space, precise estimates of the distance-from-camera can be obtained for each fish. By using the 95% percentile of the observed distance from camera to establish species-specific areas surveyed, we account for differences in detectability among species while avoiding diluting density estimates by using the maximum distance a species was observed. Accounting for this range of detectability is critical to accurately estimate fish abundances. This methodology will facilitate the integration of rotating stereo-video tools in both applied science and management contexts.

We describe a new method for counting fishes, and estimating relative abundance (MaxN) and fish density using rotating stereo-video camera systems. We also demonstrate how to use distance from camera (Z distance) to estimate species-specific detectability.

利用水下立体视频工具量化鱼类密度的新方法研究进展

The use of video camera systems in ecological studies of fish continues to gain traction as a viable, non-extractive method of measuring fish lengths and ...

科研

环境科学

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 ...

Raising the Mexican Tetra Astyanax mexicanus for Analysis of Post-larval Phenotypes and Whole-mount Immunohistochemistry

River and cave-adapted populations of Astyanax mexicanus show differences in morphology, physiology, and behavior. Research focused on comparing adult forms has revealed the genetic basis of some of these differences. Less is known about how the populations differ at post-larval stages (at the onset of feeding). Such studies may provide insight into how cavefish survive through adulthood in their natural environment. Methods for comparing post-larval development in the laboratory require standardized aquaculture and feeding regimes. Here we describe how to raise fish on a diet of nutrient-rich rotifers in non-recirculating water for up to two-weeks post fertilization. We demonstrate how to collect post-larval fish from this nursery system and perform whole-mount immunostaining. Immunostaining is an attractive alternative to transgene expression analysis for investigating development and gene function in A. mexicanus. The nursery method can also be used as a standard protocol for establishing density-matched populations for growth into adults.

Raising the Mexican Tetra Astyanax mexicanus for Analysis of Post-larval Phenotypes and Whole-mount Immunohistochemistry

In this protocol, we demonstrate how to breed Astyanax mexicanus adults, raise the larvae, and perform whole-mount immunohistochemistry on post-larval fish to compare the phenotypes of surface and cave morphotypes.

提高墨西哥龙舌兰对幼虫后表型和全体免疫组织化学的分析

River and cave-adapted populations of Astyanax mexicanus show differences in morphology, physiology, and behavior. Research focused on comparing adult ...

遗传学

Using the FishSim Animation Toolchain to Investigate Fish Behavior: A Case Study on Mate-Choice Copying In Sailfin Mollies

Over the last decade, employing computer animations for animal behavior research has increased due to its ability to non-invasively manipulate the appearance and behavior of visual stimuli, compared to manipulating live animals. Here, we present the FishSim Animation Toolchain, a software framework developed to provide researchers with an easy-to-use method for implementing 3D computer animations in behavioral experiments with fish. The toolchain offers templates to create virtual 3D stimuli of five different fish species. Stimuli are customizable in both appearance and size, based on photographs taken of live fish. Multiple stimuli can be animated by recording swimming paths in a virtual environment using a video game controller. To increase standardization of the simulated behavior, the prerecorded swimming path may be replayed with different stimuli. Multiple animations can later be organized into playlists and presented on monitors during experiments with live fish.
In a case study with sailfin mollies (Poecilia latipinna), we provide a protocol on how to conduct a mate-choice copying experiment with FishSim. We utilized this method to create and animate virtual males and virtual model females, and then presented these to live focal females in a binary choice experiment. Our results demonstrate that computer animation may be used to simulate virtual fish in a mate-choice copying experiment to investigate the role of female gravid spots as an indication of quality for a model female in mate-choice copying.
Applying this method is not limited to mate-choice copying experiments but can be used in various experimental designs. Still, its usability depends on the visual capabilities of the study species and first needs validation. Overall, computer animations offer a high degree of control and standardization in experiments and bear the potential to &#39;reduce&#39; and &#39;replace&#39; live stimulus animals as well as to &#39;refine&#39; experimental procedures.

Using the FishSim Animation Toolchain to Investigate Fish Behavior: A Case Study on Mate-Choice Copying In Sailfin Mollies

Using the novel FishSim Animation Toolchain, we present a protocol for non-invasive visual manipulation of public information in the context of mate-choice copying in sailfin mollies. FishSim Animation Toolchain provides an easy-to-use framework for the design, animation and presentation of computer-animated fish stimuli for behavioral experiments with live test fish.

利用fishsim动画工具链调查鱼类行为--以鱼翅钼中的选物复制为例

Over the last decade, employing computer animations for animal behavior research has increased due to its ability to non-invasively manipulate the ...

行为学

A Standardized Protocol for Preference Testing to Assess Fish Welfare

Animal welfare assessment techniques try to take into consideration the specific needs and wants of the animal in question. Providing enrichment (the addition of physical objects or conspecifics in the housing environment) is often a way to give captive animals the opportunity to choose who or what they interact with and how they spend their time. A fundamental component of the aquatic environment that is often overlooked in captivity, however, is the ability for the animal to choose to engage in physical exercise. For many animals, including fish, exercise is an important aspect of their life history, and is known to have many health benefits, including positive changes in the brain and behavior. Here we present a method for assessing habitat preferences in captive animals. The protocol could easily be adapted to look at a variety of environmental factors (e.g., gravel versus sand as a substrate, plastic plants versus live plants, low flow versus high flow of water) in different aquatic species, or for use with terrestrial species. Statistical assessment of preference is carried out using Jacob&#39;s preference index, which ranks the habitats from -1 (avoidance) to +1 (most preferred). With this information, it can be determined what the animal wants from a welfare perspective, including their preferred location.

A fundamental aspect of assessing the welfare of animals in captivity is to ask whether the animals have what they want. Here, we present a protocol to determine housing preference in the zebrafish (Danio rerio) with respect to the presence/absence of environmental enrichment and access to flowing of water.

偏好测试评估鱼类福利的标准化协议

Animal welfare assessment techniques try to take into consideration the specific needs and wants of the animal in question. Providing enrichment (the ...

A Novel Method of Drug Administration to Multiple Zebrafish (Danio rerio) and the Quantification of Withdrawal

Anxiety testing in zebrafish is often studied in combination with the application of pharmacological substances. In these studies, fish are routinely netted and transported between home aquaria and dosing tanks. In order to enhance the ease of compound administration, a novel method for transferring fish between tanks for drug administration was developed. Inserts that are designed for spawning were used to transfer groups of fish into the drug solution, allowing accurate dosing of all fish in the group. This increases the precision and efficiency of dosing, which becomes very important in long schedules of repeated drug administration. We implemented this procedure for use in a study examining the behavior of zebrafish in the light/dark test after administering ethanol with differing 21 day schedules. In fish exposed to daily-moderate amounts of alcohol there was a significant difference in location preference after 2 days of withdrawal when compared to the control group. However, a significant difference in location preference in a group exposed to weekly-binge administration was not observed. 
This protocol can be generalized for use with all types of compounds that are water-soluble and may be used in any situation when the behavior of fish during or after long schedules of drug administration is being examined. The light/dark test is also a valuable method of assessing withdrawal-induced changes in anxiety.

A Novel Method of Drug Administration to Multiple Zebrafish Danio rerio and the Quantification of Withdrawal

A novel method for reducing variability when exposing fish to drugs is explained. Fish exposed to various patterns of ethanol exposure were found to have altered anxiety levels during withdrawal in a light/dark scototaxic assay.

药品监督管理局多个斑马鱼的新方法（<em&gt;斑马鱼</em&gt;）和退出的量化

Anxiety testing in zebrafish is often studied in combination with the application of pharmacological substances. In these studies, fish are routinely ...

神经科学

Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is important to standardize protocols to ensure proper analyses and interpretations. In our previous studies, we have measured boldness by monitoring a series of behaviors (time near surface, latency in transitions, number of transitions, and darts) in a 1.5 L trapezoidal tank. Likewise, we quantified aggression by observing bites, lateral displays, darts, and time near an inclined mirror in a rectangular 19 L tank. By dividing a 76 L tank into thirds, we also examined shoaling preferences. The shoaling assay is a highly customizable assay and can be tailored for specific hypotheses. However, protocols for this assay also must be standardized, yet flexible enough for customization. In previous studies, end chambers were either empty, contained 5 or 10 zebrafish, or 5 pearl danios (D. albolineatus). In the following manuscript, we present a detailed protocol and representative data that accompany successful applications of the protocol, which will allow for replication of behavioral syndrome experiments.

This manuscript describes the setup, implementation, and analysis of boldness, aggression, and shoaling in zebrafish and testing for the presence of a behavioral syndrome. A standardized approach for behavioral quantification will allow for easier comparison across studies. Modifications to this protocol are possible as each assay can be run individually.

气魄，侵略和浅水测定斑马鱼行为综合征

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is ...

医学

Regular Care and Maintenance of a Zebrafish (Danio rerio) Laboratory: An Introduction

This protocol describes regular care and maintenance of a zebrafish laboratory. Zebrafish are now gaining popularity in genetics, pharmacological and behavioural research. As a vertebrate, zebrafish share considerable genetic sequence similarity with humans and are being used as an animal model for various human disease conditions. The advantages of zebrafish in comparison to other common vertebrate models include high fecundity, low maintenance cost, transparent embryos, and rapid development. Due to the spur of interest in zebrafish research, the need to establish and maintain a productive zebrafish housing facility is also increasing. Although literature is available for the maintenance of a zebrafish laboratory, a concise video protocol is lacking. This video illustrates the protocol for regular housing, feeding, breeding and raising of zebrafish larvae. This process will help researchers to understand the natural behaviour and optimal conditions of zebrafish husbandry and hence troubleshoot experimental issues that originate from the fish husbandry conditions. This protocol will be of immense help to researchers planning to establish a zebrafish laboratory, and also to graduate students who are intending to use zebrafish as an animal model.

Regular Care and Maintenance of a Zebrafish Danio rerio Laboratory: An Introduction

This protocol outlines regular maintenance and care to maintain optimal conditions for zebrafish husbandry. The video illustrates the protocol for system maintenance, regular housing, feeding, breeding, and raising of zebrafish larvae.

定期保养和维护的斑马鱼（<em&gt;斑马鱼</em&gt;）实验室简介

This protocol describes regular care and maintenance of a zebrafish laboratory. Zebrafish are now gaining popularity in genetics, pharmacological and ...

生物学

Gamete Collection and In Vitro Fertilization of Astyanax mexicanus

Astyanax mexicanus is emerging as a model organism for a variety of research fields in biological science. Part of the recent success of this teleost fish species is that it possesses interfertile cave and river-dwelling populations. This enables the genetic mapping of heritable traits that were fixed during adaptation to the different environments of these populations. While this species can be maintained and bred in the lab, it is challenging to both obtain embryos during the daytime and create hybrid embryos between strains. In vitro fertilization (IVF) has been used with a variety of different model organisms to successfully and repeatedly breed animals in the lab. In this protocol, we show how, by acclimatizing A. mexicanus to different light cycles coupled with changes in water temperature, we can shift breeding cycles to a chosen time of the day. Subsequently, we show how to identify suitable parental fish, collect healthy gametes from males and females, and produce viable offspring using IVF. This enables related procedures such as the injection of genetic constructs or developmental analysis to occur during normal working hours. Furthermore, this technique can be used to create hybrids between the cave and surface-dwelling populations, and thereby enable the study of the genetic basis of phenotypic adaptations to different environments.

Gamete Collection and In Vitro Fertilization of Astyanax mexicanus

In vitro fertilization is a commonly used technique with a variety of model organisms to maintain lab populations and produce synchronized embryos for downstream applications. Here, we present a protocol that implements this technique for different populations of the Mexican tetra fish, Astyanax mexicanus.

阿斯蒂亚纳斯墨西哥的Gamete收集与体外受精

Astyanax mexicanus is emerging as a model organism for a variety of research fields in biological science. Part of the recent success of this teleost fish ...

Sperm Collection and Computer-Assisted Sperm Analysis in the Teleost Model Japanese Medaka (Oryzias latipes)

Japanese medaka (Oryzias latipes) is a teleost fish and&#160;an emerging vertebrate model for ecotoxicology, developmental, genetics, and physiology research. Medaka is also used extensively to investigate vertebrate reproduction, which is an essential biological function as it allows a species to perpetuate. Sperm quality is an important indicator of male fertility and, thus, reproduction success. Techniques for extracting sperm and sperm analysis are well documented for many species, including teleost fish. Collecting sperm is relatively simple in larger fish but can be more complicated in small model fish as they produce less sperm and are more delicate. This article, therefore, describes two methods of sperm collection in the small model fish, Japanese medaka: testes dissection and abdominal massage. This paper demonstrates that both approaches are feasible for medaka and shows that abdominal massage can be performed a repeated number of times as the fish quickly recover from the procedure. This article also describes a protocol for computer-assisted sperm analysis in medaka to objectively assess several important indicators of medaka sperm quality (motility, progressivity, duration of motility, relative concentration). These procedures, specified for this useful small teleost model, will greatly enhance understanding of the environmental, physiological, and genetic factors influencing fertility in vertebrate males.

Sperm Collection and Computer-Assisted Sperm Analysis in the Teleost Model Japanese Medaka Oryzias latipes

This article describes two quick and efficient methods for collecting sperm from the small model fish medaka (Oryzias latipes), as well as a protocol for reliably assessing sperm quality using computer-assisted sperm analysis (CASA).

硬骨模型日本青鳉鱼（Oryzias latipes）中的精子收集和计算机辅助精子分析

Japanese medaka (Oryzias latipes) is a teleost fish and&#160;an emerging vertebrate model for ecotoxicology, developmental, genetics, and physiology ...

鱼类生殖生态学研究的基本方法

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