Name: maintaining laboratory cultures of gryllus bimaculatus, a versatile orthopteran model for insect agriculture and invertebrate physiology
Uploaded: 2022-06-08T12:30:00
Description: Watch this Scientific Journal Video about Maintaining Laboratory Cultures of Gryllus bimaculatus, a Versatile Orthopteran Model for Insect Agriculture and Invertebrate Physiology at JoVE.com

该项目的资金是通过威斯康星大学麦迪逊分校的内部赠款实现的。衷心感谢Bachhuber Consulting Inc.的Kevin Bachhuber访问他未发表的商业板球饲养指南，并感谢Michael Bartlett Smith在改进和排除这些方法方面的帮助。

1. 制备产卵底物
注意：椰子椰壳纤维是 双斑蝽的理想产卵底物。有关如何将椰壳纤维与压缩椰壳体砖分离的详细方法以及有关呼吸安全的注意事项，请参阅 补充材料 步骤1.1。
<ol><li>用肥皂和水洗手。</li>
	<li>将一个干净的容器放在天平上，称量一团大约相当于人类拳头大小的干椰子椰壳。</li>
	<li>将椰壳纤维放入可密封、干净的容器中，该?...

<ol>
	<li>Hales, K. G., Korey, C. A., Larracuente, A. M., Roberts, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetics+on+the+fly:+a+primer+on+the+Drosophila+model+system.">Genetics on the fly: a primer on the Drosophila model system.</a> Genetics. 201 (3), 815-842 (2015).</li><li>Merkel, G. <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+temperature+and+food+quality+on+the+larval+development+of+Gryllus+bimaculatus+(Orthoptera,+Gryllidae).">The effects of temperature and food quality on the larval development of Gryllus bimaculatus (Orthoptera, Gryllidae).</a> Oecologia. 30 (2), 129-140 (1977).</li><li>Bateman, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mate+preference+for+novel+partners+in+the+cricket+Gryllus+bimaculatus.">Mate preference for novel partners in the cricket Gryllus bimaculatus.</a> Ecological Entomology. 23 (4), 473-475 (1998).</li><li>Mito, T., Noji, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+two-spotted+cricket+Gryllus+bimaculatus:+An+emerging+model+for+developmental+and+regeneration+studies.">The two-spotted cricket Gryllus bimaculatus: An emerging model for developmental and regeneration studies.</a> Cold Spring Harbor Protocols. 2008, (2008).</li><li>Ylla, G., 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=Cricket+genomes:+the+genomes+of+future+food.">Cricket genomes: the genomes of future food.</a> BioRxiv. , (2020).</li><li>Halloran, A., Roos, N., Hanboonsong, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cricket+farming+as+a+livelihood+strategy+in+Thailand.">Cricket farming as a livelihood strategy in Thailand.</a> Geographical Journal. 183 (1), 112-124 (2017).</li><li>Wade, M., Hoelle, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+edible+insect+industrialization:+scales+of+production+and+implications+for+sustainability.">A review of edible insect industrialization: scales of production and implications for sustainability.</a> Environmental Research Letters. 15, 123013 (2020).</li><li>. Studies on the influence of different diets and rearing conditions on the development and growth of the two-spotted cricket Gryllus bimaculatus de Greer Available from: <a target="_blank" href="https://epub.uni-bayreuth.de/310/1/Diss.pdf">https://epub.uni-bayreuth.de/310/1/Diss.pdf</a> (2011)</li><li>Ngonga, C. A., Gor, C. O., Okuto, E. A., Ayieko, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Growth+performance+of+Acheta+domesticus+and+Gryllus+bimaculatus+production+reared+under+improvised+cage+system+for+increased+returns+and+food+security.">Growth performance of Acheta domesticus and Gryllus bimaculatus production reared under improvised cage system for increased returns and food security.</a> Journal of Insects as Food and Feed. 7, 301-310 (2021).</li><li>Behrens, W., Hoffmann, K. -. H., Kempa, S., G&#228;&#223;ler, S., Merkel-Wallner, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+diurnal+thermoperiods+and+quickly+oscillating+temperatures+on+the+development+and+reproduction+of+crickets,+Gryllus+bimaculatus.">Effects of diurnal thermoperiods and quickly oscillating temperatures on the development and reproduction of crickets, Gryllus bimaculatus.</a> Oecologia. 59 (2-3), 279-287 (1983).</li><li>Collavo, A., Paoletti, M. G., 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=Housecricket+smallscale+farming.">Housecricket smallscale farming.</a> Ecological implications of minilivestock. Potential of insects, rodents, frogs and snails. , (2005).</li><li>Simmons, L. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Competition+between+larvae+of+the+field+cricket,+Gryllus+bimaculatus+(Orthoptera:+Gryllidae)+and+its+effects+on+some+life-history+components+of+fitness.">Competition between larvae of the field cricket, Gryllus bimaculatus (Orthoptera: Gryllidae) and its effects on some life-history components of fitness.</a> Journal of Animal Ecology. 56, 1015-1027 (1987).</li><li>Sorjonen, J. M., 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+plant-based+by-product+diets+for+the+mass-rearing+of+Acheta+domesticus+and+Gryllus+bimaculatus.">The plant-based by-product diets for the mass-rearing of Acheta domesticus and Gryllus bimaculatus.</a> PLOS ONE. 14 (6), 0218830 (2019).</li><li>Maciel-Vergara, G., Jensen, A. B., Lecocq, A., Eilenberg, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diseases+in+edible+insect+rearing+systems.">Diseases in edible insect rearing systems.</a> Journal of Insects as Food and Feed. 7 (5), 1-18 (2021).</li><li>Alexander, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aggressiveness,+territoriality,+and+sexual+behavior+in+field+crickets+(Orthoptera:+Gryllidae).">Aggressiveness, territoriality, and sexual behavior in field crickets (Orthoptera: Gryllidae).</a> Behaviour. , 130-223 (1961).</li><li>Pet food, fish bait, and animal feed. USDA APHIS Available from: <a target="_blank" href="https://www.aphis.usda.gov/aphis/ourfocus/planhealth/import-information/permits/plant-pests/sa_animalfeed/ct_petfood_fishbait_animalfeed">https://www.aphis.usda.gov/aphis/ourfocus/planhealth/import-information/permits/plant-pests/sa_animalfeed/ct_petfood_fishbait_animalfeed</a> (2022)</li><li>Donoughe, S., Extavour, C. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Embryonic+development+of+the+cricket+Gryllus+bimaculatus.">Embryonic development of the cricket Gryllus bimaculatus.</a> Developmental Biology. 411 (1), 140-156 (2016).</li><li>Dobermann, D., Michaelson, L., Field, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+an+initial+high-quality+feeding+regime+on+the+survival+of+Gryllus+bimaculatus+(black+cricket)+on+bio-waste.">The effect of an initial high-quality feeding regime on the survival of Gryllus bimaculatus (black cricket) on bio-waste.</a> Journal of Insects as Food and Feed. 5 (2), 1-8 (2018).</li><li>Lundy, M. E., Parrella, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Crickets+are+not+a+free+lunch:+Protein+capture+from+scalable+organic+side-streams+via+high-density+populations+of+Acheta+domesticus.">Crickets are not a free lunch: Protein capture from scalable organic side-streams via high-density populations of Acheta domesticus.</a> PLOS ONE. 10, 0118785 (2015).</li><li>Mazaya, G., Karseno, K., Yanto, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antimicrobial+and+phytochemical+activity+of+coconut+shell+extracts.">Antimicrobial and phytochemical activity of coconut shell extracts.</a> Turkish Journal of Agriculture - Food Science and Technology. 8 (5), 1090-1097 (2020).</li></ol>

这种蟋蟀饲养方法的简单性可以使一系列研究领域受益，并代表了成功的蟋蟀饲养的通用模板，可轻松适应各种实验需求。与其他几项关于 双斑 蚴的研究相比，个体身体的成年体型较小，成熟度较慢14，我们将其归因于环境强加给我们的次优饲养温度。上述方法已在2年的时间里得到使用和改进。在商业蟋蟀养殖中有时观察到问题的证据，包括具有经典临床体征的病原体?...

以标志性昆虫果蝇 黑腹果蝇 为代表，使用昆虫作为实验室模式生物为遗传学，毒理学和生理学研究提供了明显的优势1。昆虫的小尺寸减少了培养所需的空间以及所需的饲料和消耗材料的数量。许多昆虫繁殖迅速，使它们特别适合创建专门的遗传系和需要评估多个连续世代的研究。
许多研究集中在果 蝇等全代谢昆虫上，它们表现出完全的和化蛹。但是，还有其他型号可用，包括 Grillus bimaculatus （De Geer），双斑田蟋蟀。 G. bimaculatus 是一种保尿代谢性昆虫，在达到性成熟2之前经历7至11个若虫龄。这种蟋蟀表现出与性选择相关的各种行为，包括步跖，领土展示和配偶守卫3。未成熟的蟋蟀与全代谢昆虫物种的幼虫不同，因为它们与许多保隆代谢幼虫相似，能够在蜕皮期间再生失去和受损的肢体4。此外， G. bimaculatus 的完全测序基因组于2021年发表 5.这些特征使这些蟋蟀作为基础研究的目标具有吸引力。
双斑田蟋蟀被广泛饲养，用于人类食物和动物饲料。这些手术的规模通常比实验室研究的规模大得多6，7。尽管规模不同，但研究人员面临的挑战与商业板球养殖者面临的挑战有很大重叠。这些考虑因素集中在基于实验室的研究的背景下，旨在改善食用昆虫的生产。随着食用昆虫产业的不断发展壮大，优化饲料投入和生产的其他方面是一个主要目标8。实验室研究表明，这些蟋蟀的饲养效率，存活率或生成时间都有所提高，这有可能有助于提高蟋蟀养殖业务的长期盈利能力。
标准化的饲养方案可以在研究饲养优化的研究之间进行更密切的比较。迄今为止，在实验室中饲养G. bimaculatus的深入方案很少发表。理想的协议将反映现实世界中板球养殖作业中遇到的条件，同时保持严格控制的条件，以准确测量实验处理引起的生长性能变化，并突出风险缓解策略。本文中描述的方法是基于已发表的协议，技术和设备开发的，用于在广泛的实验室和商业生产规模2，9，10，11，12中饲养各种蟋蟀物种。这些方法也得到了几个非同行评审来源的信息，包括未发表的技术公告和与北美商业板球农民的个人交流。该方案的开发旨在促进建立专门用于昆虫农业相关试验的双斑蚴实验室培养物。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>31-qt (29.3 L) Snap-lid tote bin with lid</td>
			<td>HOMZ</td>
			<td>3430CLBL</td>
			<td>Used to house breeding stock</td>
		</tr>
		<tr>
			<td>3-tier/12-tray Grow Light Stand</td>
			<td>Fischer Scientific</td>
			<td>NC1938548</td>
			<td></td>
		</tr>
		<tr>
			<td>50-gal (189.27L) tote bin with lid</td>
			<td>Sterilite</td>
			<td>#14796603</td>
			<td>Used as secondary containment when handling crickets</td>
		</tr>
		<tr>
			<td>50 mL polypropylene graduated cylinder</td>
			<td>Fischer Scientific</td>
			<td>S95171</td>
			<td></td>
		</tr>
		<tr>
			<td>7.5-qt (7.1 L) snap-lid tote bin with lid</td>
			<td>HOMZ</td>
			<td>3410CLBL</td>
			<td>Used to house exprimental stock</td>
		</tr>
		<tr>
			<td>Accuris 500 g x 0.01 g Balance</td>
			<td>Manufactured by Accuris, a subsidieary of Benchmark Scientific</td>
			<td>W3300-500</td>
			<td>Purchased from Dot Scientific through University of Wisconsin system purchasing service &#34;ShopUW+&#34;</td>
		</tr>
		<tr>
			<td>Ace Premier 1 Inch Flat Chip Brush</td>
			<td>Ace Hardware&#160;</td>
			<td>#1803261</td>
			<td></td>
		</tr>
		<tr>
			<td>Bel-Art SP Scienceware deionized water wash bottle</td>
			<td>Fischer Scientific</td>
			<td>03-421-160&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Bright aluminum window screen&#160;</td>
			<td>Phifer</td>
			<td>UNSPSC# 11162108</td>
			<td>Mesh size 18 x 16&#34;</td>
		</tr>
		<tr>
			<td>Clear Disposable Plastic Portion Cups 5.5 oz w/ lids</td>
			<td>Wal-Mart</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Deionized water</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Diablo 4-4/8&#34; x 13 TPI Ultra Fine Finish Bi-Metal Jigsaw Blade</td>
			<td>Home Depot</td>
			<td>#313114935</td>
			<td></td>
		</tr>
		<tr>
			<td>Egg Filler Flats-Paper, 12 x 12&#34;</td>
			<td>Uline</td>
			<td>S-5189</td>
			<td></td>
		</tr>
		<tr>
			<td>Fisherbrand Petri Dishes with Clear Lid 100 x 15mm</td>
			<td>Fischer Scientific</td>
			<td>FB0875714</td>
			<td></td>
		</tr>
		<tr>
			<td>Fisherbrand Petri Dishes with Clear Lid 60 x 15mm</td>
			<td>Fischer Scientific</td>
			<td>FB0875713A</td>
			<td></td>
		</tr>
		<tr>
			<td>Georgia-Pacific Envision Brown Paper Towels</td>
			<td>Home Depot</td>
			<td>#205675843</td>
			<td></td>
		</tr>
		<tr>
			<td>Infinity Tough Guy high performance hot-melt glue sticks</td>
			<td>Infinity Bond</td>
			<td>Infinity IM-Tough-Guy-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Mazuri Cricket Diet</td>
			<td>Land O&#39; Lakes International</td>
			<td>SKU#&#160; 3002219-105</td>
			<td></td>
		</tr>
		<tr>
			<td>Stanley TimeIt Twin 2-outlet Grounded Mechanical 24 Hour Timer</td>
			<td>Wal-Mart</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Vermont Organics Reclamation Soil 11 lb Coir Block</td>
			<td>Home Depot</td>
			<td>#300679904</td>
			<td></td>
		</tr>
	</tbody>
</table>

maintaining laboratory cultures of gryllus bimaculatus, a versatile orthopteran model for insect agriculture and invertebrate physiology

Gryllus bimaculatus （De Geer）是一种大型蟋蟀，分布在非洲和欧亚大陆南部，在那里它经常被野生收获作为人类食物。在其原生范围之外，由于其膳食可塑性，快速繁殖周期，缺乏隔膜要求，对高密度饲养的耐受性以及对病原体的鲁棒性，培养 G. bimaculatus 是可行的。因此， G. bimaculatus 可以成为昆虫生理学，行为学，胚胎学或遗传学研究的多功能模型。
养殖参数，如放养密度、笼内避风、光周期、温度、相对湿度和饮食，都会影响蟋蟀的生长、行为和基因表达，应予以标准化。在关于供人类消费的农业昆虫的新兴文献中，这些蟋蟀经常被用来评估来自作物残茬，食品加工副产品和其他低成本废物流的候选饲料混合物。
为了支持正在进行的评估 双斑蝽 生长性能和营养质量以响应可变饲料基质的实验，开发了一套全面的标准方案，用于实验室中的育种，保养，处理，测量和安乐死，并在此处介绍。行业标准的蟋蟀饲料已被证明营养充足，功能上适合长期维持蟋蟀种畜，以及用作实验性对照饲料。在12光（L）/ 12暗（D）光周期下， 在屏顶29.3升聚乙烯笼中，以0.005只蟋蟀/厘米3的密度饲养这些蟋蟀，平均温度为27°C，湿润的椰子椰壳既可作为水合源，又能作为产卵培养基，在2年的时间里成功地维持了健康的蟋蟀。按照这些方法，蟋蟀在对照实验中收获时的平均质量为0.724克，收获时的平均质量为0.190克，在放养（22天）和收获（65天）之间有89%的存活率和68.2%的性成熟度。

在2021年9月的饲料试验中收集了证明从孵化到65天龄的蟋蟀成功饲养的数据。按照这些方案的步骤1.1.1-2.6.1，从卵中生长蟋蟀，在6个重复的笼子里放养24只随机的22天大（三龄）蟋蟀，遵循上述步骤2.7。然后，在环境室条件下饲养蟋蟀;然而，由于设施空气处理装置故障，在20%相对湿度下，平均室温为25±1°C，而不是建议的27°C。 在孵化后22至65天之间每周测量两次蟋蟀质量。该实验的结果概述如下，?...

Watch this Scientific Journal Video about Maintaining Laboratory Cultures of Gryllus bimaculatus, a Versatile Orthopteran Model for Insect Agriculture and Invertebrate Physiology at JoVE.com

Maintaining Laboratory Cultures of Gryllus bimaculatus, a Versatile Orthopteran Model for Insect Agriculture and Invertebrate Physiology

本文概述了标准化重要因素的基本方法，例如密度，饲料可用性，水合作用来源和环境控制，用于长期饲养可食用蟋蟀 Grylus bimaculatus的实验室培养物。

维持 双斑蝽螈的实验室培养物，这是一种用于昆虫农业和无脊椎动物生理学的多功能正翅目动物模型

Gryllus bimaculatus (De Geer)&#160;is a large-bodied cricket distributed throughout Africa and Southern Eurasia where it is often wild-harvested as human ...

这些技术描绘了饲养Grylus bimaculatus的实用和经过验证的方法，Gryllus bimaculatus是一种用于动物饲料和人类食物的关键迷你牲畜物种，也是越来越重要的替代模式生物。这些用于培养Grylus bimaculatus的实验室方法源自整个北美蟋蟀养殖业中常见的技术，代表了蟋蟀养殖作业的实验室规模的传真。首先，将一个干净的容器放在天平上，并称量一团大约相当于人类拳头大小的干椰子椰壳。然后，将椰壳纤维放入可密封的清洁容器中，该容器可容纳高达原始体积六倍的膨胀。用干净的手，轻轻地从从较大块中取出的那块上轻轻地分解一团的椰壳纤维。接下来，使用250毫升量程圆柱体，测量正确体积的去离子水，以达到五份水与一份干椰壳纤维质量的五比一比例，并缓慢加入被测得的去离子水，均匀地使所有椰壳纤维颗粒水。之后，手动浸渍团块以确保均匀的水合作用，并重新去皮先前称重椰壳的容器。称出75克湿润的椰壳纤维，并使用干净的塑料勺将其转移到100毫米×15毫米的培养皿中，以确保椰壳均匀地分布在培养皿的底部并且没有团块。然后，用实验室胶带标记培养皿的侧面，并标明出生菌落和卵子收集事件的日期。将另外45毫升去离子水测量到量筒中，并在培养皿中包装的椰壳膜表面上均匀地加水，以确保均匀的水合作用。培养皿包装完成后，将剩余的湿椰壳纤维密封在气密容器中，以便在零下20摄氏度储存。将水合产卵基质放入包含所需蟋蟀亲本的笼子中，尽可能远离饲料，并记录日期和时间。然后，在工作台面上放置一个可高温高压灭菌的小垃圾容器。接下来，在垃圾箱旁边的长凳上放置一个干净的，空的，29.3升的塑料笼。将另一个装有母蟋蟀和产卵基质的29.3升笼子放在空笼子的垃圾容器的另一侧。24小时后，从母蟋蟀笼中取出产卵基质，并将其定位在可高压灭菌的废物容器上。检查产卵基材的顶部是否有蟋蟀可能踢到椰壳表面的任何碎屑或饲料颗粒。接下来，用干净的Scoopula或塑料勺将椰壳纤维污染物去除到废物容器中，并将塑料勺放入废物容器中。然后，将清洁的产卵基材放在干净的29.3升笼子中。之后，将笼子放在12小时黑暗，12小时光照光周期的培养箱中，在60%相对湿度下设置为27摄氏度。将装有种畜的笼子放回原始位置，并从工作台面上清除所有物品。然后，将废物容器放入设施内的冰箱中，专用于储存可能被蟋蟀蛋污染的物品。接下来，用10%漂白剂溶液消毒工作台面，并静置60秒。用干净的纸巾擦干工作表面。打开冰箱后，将纸巾丢弃在废物容器中。选择两个未使用的 30.8 厘米 x 30.8 厘米商用鸡蛋纸盒。用美工刀或强力剪刀将它们切成六条尺寸相等的10.1厘米宽的独立条带。然后，用手刷掉切边，去除纸板上悬垂的颗粒。接下来，将六个单独的10.1厘米×30.8厘米纸箱垂直放入笼子的底部，纸箱的较长轴跨越29.3升笼子的较窄水平轴。将第七块纸箱平放在六个直立件的顶部。选择三条原石后，将大约25厘米×25厘米的牛皮纸巾折成两半。然后，放置两个，使它们覆盖纸箱结构的近端的顶部，并将一个放在远端侧的纸箱堆叠上。产卵后第11天，将产卵底物移入笼子的近端右上角。当观察到舱口时，将放置在纸箱顶部的纸巾弄雾，直到它们被润湿，但不会主动脱落水。然后，用70%乙醇彻底擦拭100毫米培养皿盖的两侧，并使其干燥。接下来，将 50 克进料舀入 60 瓦的单份搅拌机中，以 10，000 rpm 的速度研磨一分钟。测量一克饲料后，将其摇到笼子里的培养皿盖上。使用勺子或勺子的干净端，将食物尽可能均匀地摊开在盘子的底部。之后，每两天更换一次饲料。将旧饲料丢弃在可高温高压灭菌的废物容器中。监测饲料上的霉菌生长。如果饲料开始变白或变绿，丢弃培养皿并立即喂食。产卵后14天，使用2.54厘米的画笔将所有蟋蟀从椰壳表面和培养皿侧面刷入笼中，以去除附着在出生椰壳纤维基质上的蟋蟀。接下来，将除去的产卵基质放入可高温高压灭菌的废物容器中，并将其储存在冰箱中直至高压灭菌。然后，用新鲜的椰壳纤维皿替换出生基质以进行补水。使用去离子水洗瓶，加水直至椰壳膜表面闪闪发光，但不结块。蟋蟀来自平均质量为21毫克的种群。在实验结束时，所有成年蟋蟀和成年蟋蟀的平均质量为0.724克。在65天大时，当实验结束时，有30名成年男性在场，平均质量为0.721克，而对于58名成年女性，平均质量为0.841克。放养和收获之间的存活率为89%，每周通过所有笼子中所有蟋蟀的总数来衡量。在第65天，大约68%的蟋蟀已经达到成年幼年。浇水或雾化时，避免在外壳底部形成水滴。将纸巾放在避难所上可以产生湿度梯度，并吸收雾化产生的水滴。该程序可作为许多生命科学领域的研究人员将Grylus bimaculatus作为模式生物饲养的模板。迄今为止，几乎没有关于饲养双斑鳃鳗或任何其他保尿代谢样生物的正式方案发表。编纂和标准化昆虫农业中使用的昆虫饲养方法将更好地比较研究小组之间的结果。

Research

JoVE Journal

Biology

Maintaining Laboratory Cultures of Gryllus bimaculatus, a Versatile Orthopteran Model for Insect Agriculture and Invertebrate Physiology

An Experimental and Bioinformatics Protocol for RNA-seq Analyses of Photoperiodic Diapause in the Asian Tiger Mosquito, Aedes albopictus

An Experimental and Bioinformatics Protocol for RNA-seq Analyses of Photoperiodic Diapause in the Asian Tiger Mosquito, Aedes albopictus

实验和生物信息学议定书光周期滞育的亚洲虎蚊的RNA序列分析，<em&gt;白纹伊蚊</em

Photoperiodic diapause is an important adaptation that allows individuals to escape harsh seasonal environments via a series of physiological changes, ...

科研

生物学

Physiological Recordings and RNA Sequencing of the Gustatory Appendages of the Yellow-fever Mosquito Aedes aegypti

Physiological Recordings and RNA Sequencing of the Gustatory Appendages of the Yellow-fever Mosquito Aedes aegypti

生理录音和黄热病蚊子的味觉附属物RNA测序<em&gt;埃及伊蚊</em

Electrophysiological recording of action potentials from sensory neurons of mosquitoes provides investigators a glimpse into the chemical perception of ...

Transient Expression and Cellular Localization of Recombinant Proteins in Cultured Insect Cells

瞬时表达和重组蛋白的细胞定位在培养的昆虫细胞

Heterologous protein expression systems are used for the production of recombinant proteins, the interpretation of cellular trafficking/localization, and ...

Methods for the Extraction of Endosymbionts from the Whitefly Bemisia tabaci

Methods for the Extraction of Endosymbionts from the Whitefly Bemisia tabaci

从白蛉提取内分泌物的方法<em&gt; Bemisia tabaci</em

Bacterial symbionts form an intimate relationship with their hosts and confer advantages to the hosts in most cases. Genomic information is critical to ...

Experimental Protocol for Using Drosophila As an Invertebrate Model System for Toxicity Testing in the Laboratory

Experimental Protocol for Using Drosophila As an Invertebrate Model System for Toxicity Testing in the Laboratory

利用果蝇作为无脊椎动物毒性试验模型系统的实验协议

Emergent properties and external factors (population-level and ecosystem-level interactions, in particular) play important roles in mediating ...

A Versatile Model of Hard Tick Infestation on Laboratory Rabbits

实验家兔硬蜱的多用途模型

The use of live animals in tick research is crucial for a variety of experimental purposes including the maintenance of hard tick colonies in the ...

Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A (EYFP-CENP-A)

Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A EYFP-CENP-A

质谱分析，用于识别 EYFP 标记 CENP-A （EYFP-CENP-A） 的泛化

Studying the structure and the dynamics of kinetochores and centromeres is important in understanding chromosomal instability (CIN) and cancer ...

Preparing Irradiated and Marked Male Aedes aegypti Mosquitoes for Release in an Operational Sterile Insect Technique Program

Preparing Irradiated and Marked Male Aedes aegypti Mosquitoes for Release in an Operational Sterile Insect Technique Program

准备辐照和标记的雄性 埃及伊蚊 ，以便在操作性昆虫技术计划中释放

The control of such human diseases as dengue, Zika, and chikungunya relies on the control of their vector, the Aedes aegypti mosquito, because there is no ...

Immunofluorescent Labeling of Plant Virus and Insect Vector Proteins in Hemipteran Guts

半翅目肠道中植物病毒和昆虫载体蛋白的免疫荧光标记

Most plant viruses in nature are transmitted from one plant to another by hemipteran insects. A high population density of the vector insects that are ...

维持 双斑蝽螈的实验室培养物，这是一种用于昆虫农业和无脊椎动物生理学的多功能正翅目动物模型

维持双斑蝽螈的实验室培养物，这是一种用于昆虫农业和无脊椎动物生理学的多功能正翅目动物模型