Name: methods for rearing the parasitoid ganaspis brasiliensis, a promising biological control agent for the invasive drosophila suzukii
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Description: Watch this Scientific Journal Video about Methods for Rearing the Parasitoid Ganaspis brasiliensis, a Promising Biological Control Agent for the Invasive Drosophila suzukii at JoVE.com

作者感谢卢卡斯·西豪森和马克·肯尼斯（CABI，瑞士）善意地提供了G1 G. 巴西。意大利的资金由意大利特伦托的特伦托自治省提供，在美国由国家粮食及农业研究所提供，美国农业部特种作物研究计划奖（#2020-5118-32140），美国农业部动植物卫生检查局（农业法案，基金14-8130-0463）和美国农业部ARS CRIS基本基金（项目8010-22000-033-00D）。美国农业部是机会均等的提供者和雇主，不认可本出版物中提到的产品。

1. 巴西G3甘草小规模实验室饲养方法
<ol><li>准备主人的饮食。<ol>
<li>将600 mL蒸馏水加入1，500 mL玻璃容器中，并在热板上加热水。</li>
		<li>加入88.6克市售干粮（由琼脂，啤酒酵母，玉米粉，对羟基苯甲酸甲酯和蔗糖制成）或使用道尔顿等人发表的配方准备饮食28 （见步骤2.1.2）。</li>
		<li>在干粮中加入300毫升蒸馏水，并彻底搅拌饮食混合物。</li>
		<li>将混合物...

<ol>
	<li>Asplen, M. 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=Invasion+biology+of+spotted+wing+drosophila+(Drosophila+suzukii):+a+global+perspective+and+future+priorities.">Invasion biology of spotted wing drosophila (Drosophila suzukii): a global perspective and future priorities.</a> Journal of Pest Science. 88 (3), 469-494 (2015).</li><li>Tait, 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=Drosophila+suzukii+(Diptera:+Drosophilidae):+A+decade+of+research+towards+a+sustainable+integrated+pest+management+program.">Drosophila suzukii (Diptera: Drosophilidae): A decade of research towards a sustainable integrated pest management program.</a> Journal of Economic Entomology. 114 (5), 1950-1974 (2021).</li><li>Kirschbaum, D. S., Funes, C. F., Buonocore-Biancheri, M. 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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=Biological+control+of+spotted-wing+drosophila+(Diptera:+Drosophilidae):+Current+and+pending+tactics.">Biological control of spotted-wing drosophila (Diptera: Drosophilidae): Current and pending tactics.</a> Journal of Integreated Pest Management. 10 (1), 13 (2019).</li><li>Wang, X. G., Lee, J. C., Daane, K. M., Hoelmer, K. A., Garcia, F. R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biological+control+of+spotted-wing+drosophila:+An+update+on+promising+agents.">Biological control of spotted-wing drosophila: An update on promising agents.</a> Drosophilia suzukii management. , 143-167 (2020).</li><li>Daane, K. 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=First+exploration+of+parasitoids+of+Drosophila+suzukii+in+South+Korea+as+potential+classical+biological+agents.">First exploration of parasitoids of Drosophila suzukii in South Korea as potential classical biological agents.</a> Journal of Pest Science. 89 (3), 823-835 (2016).</li><li>Giorgini, 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=Exploration+for+native+parasitoids+of+Drosophila+suzukii+in+China+reveals+a+diversity+of+parasitoid+species+and+narrow+host+range+of+the+dominant+parasitoid.">Exploration for native parasitoids of Drosophila suzukii in China reveals a diversity of parasitoid species and narrow host range of the dominant parasitoid.</a> Journal of Pest Science. 92 (2), 509-522 (2019).</li><li>Girod, P., 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+parasitoid+complex+of+D.+suzukii+and+other+fruit+feeding+Drosophila+species+in+Asia.">The parasitoid complex of D. suzukii and other fruit feeding Drosophila species in Asia.</a> Scientific Reports. 8 (1), 11839 (2018).</li><li>Kacsoh, B. Z., Schlenke, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+hemocyte+load+is+associated+with+increased+resistance+against+parasitoids+in+Drosophila+suzukii,+a+relative+of+D.+melanogaster.">High hemocyte load is associated with increased resistance against parasitoids in Drosophila suzukii, a relative of D. melanogaster.</a> PLoS ONE. 7 (4), 34721 (2012).</li><li>Buffington, M. L., Forshage, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Redescription+of+Ganaspis+brasiliensis+(Ihering,+1905),+new+combination+(Hymenoptera:+Figitidae),+a+natural+enemy+of+the+invasive+Drosophila+suzukii+(Matsumura,+1931)(Diptera:+Drosophilidae).">Redescription of Ganaspis brasiliensis (Ihering, 1905), new combination (Hymenoptera: Figitidae), a natural enemy of the invasive Drosophila suzukii (Matsumura, 1931)(Diptera: Drosophilidae).</a> Procedings of the Entomoogical Society of Washington. 118 (1), 1-13 (2016).</li><li>Biondi, A., 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=Innate+olfactory+responses+of+Asobara+japonica+(Hymenoptera:+Braconidae)towards+fruits+infested+by+the+invasive+spotted+wing+drosophila.">Innate olfactory responses of Asobara japonica (Hymenoptera: Braconidae)towards fruits infested by the invasive spotted wing drosophila.</a> Journal of Insect Behavior. 30 (5), 495-506 (2017).</li><li>Biondi, A., Wang, X. G., Daane, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Host+preference+of+three+Asian+larval+parasitoids+to+closely+related+Drosophila+species:+implications+for+biological+control+of+Drosophila+suzukii.">Host preference of three Asian larval parasitoids to closely related Drosophila species: implications for biological control of Drosophila suzukii.</a> Journal of Pest Science. 94 (2), 273-283 (2021).</li><li>Girod, P., Rossignaud, L., Haye, T., Turlings, T. C. J., Kenis, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+Asian+parasitoids+in+larvae+of+Drosophila+suzukii+feeding+on+blueberry+and+artificial+diet.">Development of Asian parasitoids in larvae of Drosophila suzukii feeding on blueberry and artificial diet.</a> Journal of Applied Entomology. 142 (5), 483-494 (2018).</li><li>Wang, X. G., Nance, A. H., Jones, J. M. L., Hoelmer, K. A., Daane, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aspects+of+the+biology+and+reproductive+strategy+of+two+Asian+larval+parasitoids+evaluated+for+classical+biological+control+of+Drosophila+suzukii.">Aspects of the biology and reproductive strategy of two Asian larval parasitoids evaluated for classical biological control of Drosophila suzukii.</a> Biological Control. 121, 58-65 (2018).</li><li>Wang, X. G., Biondi, A., Daane, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+responses+of+three+candidate+Asian+larval+parasitoids+evaluated+for+classical+biological+control+of+Drosophila+suzukii.">Functional responses of three candidate Asian larval parasitoids evaluated for classical biological control of Drosophila suzukii.</a> Journal of Economic Entomology. 113 (1), 73-80 (2020).</li><li>Wang, X. 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=Assessment+of+Asobara+japonica+as+a+potential+biological+control+agent+for+the+spotted+wing+drosophila,+Drosophila+suzukii.">Assessment of Asobara japonica as a potential biological control agent for the spotted wing drosophila, Drosophila suzukii.</a> Entomologia Generalis. 41, 1-12 (2021).</li><li>Hougardy, E., Hogg, B. N., Wang, X. G., Daane, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+thermal+performances+of+two+Asian+larval+parasitoids+of+Drosophila+suzukii.">Comparison of thermal performances of two Asian larval parasitoids of Drosophila suzukii.</a> Biological Control. 136, 104000 (2019).</li><li>Wang, X. G., Hogg, B. N., Hougardy, E., Nance, A. H., Daane, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potential+competitive+outcomes+among+three+solitary+larval+endoparasitoids+as+candidate+agents+for+classical+biological+control+of+Drosophila+suzukii.">Potential competitive outcomes among three solitary larval endoparasitoids as candidate agents for classical biological control of Drosophila suzukii.</a> Biological Control. 130, 18-26 (2019).</li><li>Daane, K. M., Biondi, A., Wang, X. G., Hogg, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potential+host+ranges+of+three+Asian+larval+parasitoids+of+Drosophila+suzukii.">Potential host ranges of three Asian larval parasitoids of Drosophila suzukii.</a> Journal of Pest Science. 94 (4), 1171-1182 (2021).</li><li>Girod, P., 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=Host+specificity+of+Asian+parasitoids+for+potential+classical+biological+control+of+Drosophila+suzukii.">Host specificity of Asian parasitoids for potential classical biological control of Drosophila suzukii.</a> Journal of Pest Science. 91 (4), 1241-1250 (2018).</li><li>Seehausen, M. L., 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=Evidence+for+a+cryptic+parasitoid+species+reveals+its+suitability+as+a+biological+control+agent.">Evidence for a cryptic parasitoid species reveals its suitability as a biological control agent.</a> Scientific Reports. 10 (1), 19096 (2020).</li><li>Nomano, F. Y., 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=Genetic+differentiation+of+Ganaspis+brasiliensis+(Hymenoptera:+Figitidae)+from+East+and+Southeast+Asia.">Genetic differentiation of Ganaspis brasiliensis (Hymenoptera: Figitidae) from East and Southeast Asia.</a> Applied Entomology and Zoology. 52 (3), 429-437 (2017).</li><li>Kasuya, N., Mitsui, H., Ideo, S., Watada, M., Kimura, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ecological,+morphological+and+molecular+studies+on+Ganaspis+individuals+(Hymenoptera:+Figitidae)+attacking+Drosophila+suzukii+(Diptera:+Drosophilidae).">Ecological, morphological and molecular studies on Ganaspis individuals (Hymenoptera: Figitidae) attacking Drosophila suzukii (Diptera: Drosophilidae).</a> Applied Entomology and Zoology. 48 (1), 87-92 (2013).</li><li>Matsuura, A., Mitsui, H., Kimura, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+preliminary+study+on+distributions+and+oviposition+sites+of+Drosophila+suzukii+(Diptera:+Drosophilidae)+and+its+parasitoids+on+wild+cherry+tree+in+Tokyo,+central+Japan.">A preliminary study on distributions and oviposition sites of Drosophila suzukii (Diptera: Drosophilidae) and its parasitoids on wild cherry tree in Tokyo, central Japan.</a> Applied Entomology and Zoology. 53 (1), 47-53 (2018).</li><li>Abram, P. 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=New+records+of+Leptopilina,+Ganaspis,+and+Asobara+species+associated+with+Drosophila+suzukii+in+North+America,+including+detections+of+L.+japonica+and+G.+brasiliensis.">New records of Leptopilina, Ganaspis, and Asobara species associated with Drosophila suzukii in North America, including detections of L. japonica and G. brasiliensis.</a> Journal of Hymenoptera Research. 78, 1-17 (2020).</li><li>Puppato, S., Grassi, A., Pedrazzoli, F., De Cristofaro, A., Ioriatti, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=First+report+of+Leptopilina+japonica+in+Europe.">First report of Leptopilina japonica in Europe.</a> Insects. 11 (9), 611 (2020).</li><li>Dalton, D. T., 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=Laboratory+survival+of+Drosophila+suzukii+under+simulated+winter+conditions+of+the+Pacific+Northwest+and+seasonal+field+trapping+in+five+primary+regions+of+small+and+stone+fruit+production+in+the+United+States.">Laboratory survival of Drosophila suzukii under simulated winter conditions of the Pacific Northwest and seasonal field trapping in five primary regions of small and stone fruit production in the United States.</a> Pest Managagement Science. 67 (11), 1368-1374 (2011).</li></ol>

生物控制剂的长期研究和随后的田间释放取决于有效和经济的饲养技术的可用性。本研究中描述的方法已被证明是小规模和大规模饲养 巴西加纳斯皮斯的有效方案。小规模饲养方案已经开发了几年，以优化劳动力并减少同时维持寄生虫和宿主菌落所需的专用设备。它适用于维持实验室研究或生物测定的菌落。作者已经使用类似的方法来饲养这种寄生虫，用于隔离评估这种寄生虫。大规模饲...

原产于东亚的斑点翅果蝇，铃木果蝇（松村果蝇）（双翅目：果蝇科），已在美洲，欧洲和非洲部分地区广泛建立1，2。苍蝇是极其多食性的，能够利用其原生和入侵地区1，2，3中各种具有柔软和薄皮的栽培和野生水果。目前对这种害虫的管理策略严重依赖于在易感水果成熟时频繁使用针对农田中成年苍蝇的杀虫剂。经常使用重复喷洒，可能是由于水库苍蝇种群从非作物栖息地不断溢出，以及缺乏居住在被入侵地区的有效天敌1，4。生物控制，特别是通过自我延续的特化寄生虫，可能有助于在景观水平上抑制苍蝇种群，并在可持续地管理这种高度移动性和多食性害虫的全区域范围内发挥关键作用4，5，6。
在过去的十年中，研究人员一直致力于在东亚7，8，9的苍蝇原生范围内发现铃木果蝇的共同进化寄生虫，以及在美洲和欧洲4，5，6的苍蝇入侵地区有效但新关联的寄生虫。在苍蝇新入侵的地区，常见的幼虫果蝇寄生虫，如阿索巴拉c.f.塔比达（Nees）（膜翅目：布拉科），胭脂虫（巴博廷等人）和异体乳杆菌（汤普森）（膜翅目：Figitidae），由于苍蝇的强烈免疫抵抗力，无法从铃木上的寄生虫发育或具有低寄生虫水平。只有一些世界性和通才的蛹类寄生虫，如长春花（Rondani）（膜翅目：翼手目）和果蝇（Perkins）（膜翅目：Diapriidae），以及南美洲的利马毛滴虫可以很容易地从这种苍蝇中发展出来 4.相比之下，在东亚的探索已经从铃木4，5，6中发现了许多幼虫寄生虫。其中，阿索巴拉粳稻贝洛科贝利斯基，巴西加那斯皮斯和日本粳稻诺夫科维奇和木村幼虫是主要的幼虫寄生虫7，8，9，11。特别是，两种无花果体（粳稻和巴西猪笼草）是主要在天然植被中由铃木猪笼草和/或其他密切相关的果蝇感染的新鲜水果中发现的主要寄生虫7，8，9。这三种亚洲幼虫寄生虫被输入到美国和欧洲的检疫设施，并评估了其相对效率12，13，14，15，16，17，气候适应性18，潜在的种间竞争相互作用19，最重要的是宿主特异性8，20，21，22.
检疫评估显示，与其他测试的亚洲幼虫寄生虫相比，巴西果蝇对铃木果蝇的宿主特异性更高，尽管它可能由不同的生物型或具有不同宿主特异性的神秘物种组成8，21，22，23，24。Nomano等人根据对线粒体细胞色素氧化酶I基因片段的分子分析，将来自不同地理区域的Ganaspis个体分为五个遗传组（命名为G1-G5）。据报道，G2 和 G4 组仅来自少数南亚热带地区，而 G5 组的报告来自亚洲和其他地区（例如阿根廷、巴西、夏威夷和墨西哥），其宿主不明（Buffington，个人观察）。在韩国7，中国8和日本9，23，25中，铃木猪笼草侵扰的野生水果的野外收集物单独发现或代表G1和G3组的标本混合物。这两个群体似乎是对称的，并且共存于铃木猪和其他密切相关的寄主蝇居住的同一寄主植物上。尽管如此，两组之间已经观察到一些差异，G1似乎比G3对铃木D.具有更高程度的宿主或宿主栖息地特异性，尽管它们在检疫测试21，22中都攻击了许多密切相关的物种。进一步详细的分子分析可能有助于确定物种状态，特别是对于G1和G3组。这项研究将它们称为G1 G.巴西菌和G3 G. 巴西菌。一些早期的研究也将G1 G. 巴西莲命名为G。 参见巴西14，21，22。G1 G. 巴西蜥蜴最近被批准在美国和意大利（其他几个欧洲国家目前正在考虑引入）对铃木乌贼进行田间释放，而G3巴西鳞毛蕨可能考虑在不久的将来进行田间释放。最近的调查还发现，在加拿大不列颠哥伦比亚省26和美国华盛顿州，粳稻和G1 G. Brasiliensis的偶然种群（Beers等人，未发表的数据），以及意大利特伦托省的粳稻柳叶杆菌种群27。
鉴于对制定 铃木果蝇 管理的生物控制方案的极大兴趣，以及偶然和故意引入 巴西果蝇的巨大生物控制潜力，有必要为这种幼虫寄生虫开发有效的饲养方法，以便今后进行长期研究和/或田间释放。该协议和相关视频文章描述了这种寄生虫的两套饲养方法：（1）使用宿主果实（蓝莓）和人造饲料的混合物在烧瓶中进行小规模实验室饲养，以培养 铃木杜鹃。这些方法是使用最初从中国昆明收集的G3材料开发的。（2）使用寄主果实（蓝莓）进行大规模养殖，以便在大型网箱中放牧 ，用于铃木杜鹃的养殖。用于大规模养殖的遗传组是原产于日本东京的G1种群9，22。还简要讨论了其他规模的饲养方法，例如为两组使用小瓶或小容器。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Active dry yeast</td>
			<td>Fleischmanns Yeast, Cincinatti, OH, USA</td>
			<td>None</td>
			<td>Used to cover fruit to reduce mold growth and enhance the frui attraction to the flies</td>
		</tr>
		<tr>
			<td>Bacteriological agar</td>
			<td>Merk Life Science S.r.l., Milan, Italy</td>
			<td>A1296 - 5KG</td>
			<td>Used to prepare the Standard Drosophila Medium</td>
		</tr>
		<tr>
			<td>Bleach solution</td>
			<td>Clorox Company, Oakland, CA, USA</td>
			<td>None</td>
			<td>Used to disinfect flesh fruit</td>
		</tr>
		<tr>
			<td>Blue stopper</td>
			<td>Azer Scientific, Morgantown, PA, USA</td>
			<td>ES3837</td>
			<td>Used for sealing the tube while allowing ventilation for insects</td>
		</tr>
		<tr>
			<td>Blueberries</td>
			<td>Grocery Store, Newark, DE, USA</td>
			<td>None</td>
			<td>Provided as host fruit for the flies (various other fruit can also be used)</td>
		</tr>
		<tr>
			<td>BugDorm insect rearing cage (W24.5 x D24.5 x H63.0 cm)</td>
			<td>Mega View Science Co. Ltd., Taichung, Taiwan</td>
			<td>4E3030</td>
			<td>Used for rearing parasitoids (parasitism cage)</td>
		</tr>
		<tr>
			<td>BugDorm insect rearing cage (W32.5 x D32.5 x H32.5 cm)</td>
			<td>Mega View Science Co. Ltd., Taichung, Taiwan</td>
			<td>4E4590</td>
			<td>Used for rearing flies</td>
		</tr>
		<tr>
			<td>BugDorm insect rearing cage (W32.5 x D32.5 x H32.5 cm)</td>
			<td>Mega View Science Co. Ltd., Taichung, Taiwan</td>
			<td>4E4545</td>
			<td>Used for rearing parasitoids (eclosion cage)</td>
		</tr>
		<tr>
			<td>Chicken wire (0.64 cm, 19 gauge)</td>
			<td>Everbilt, OH, USA</td>
			<td>308231EB</td>
			<td>Used to lift up the fruit to allow maximum parasitoid oviposition</td>
		</tr>
		<tr>
			<td>Cornmeal</td>
			<td>Grocery Store, Trento, TN, Italy</td>
			<td>None</td>
			<td>Used to prepare the Standard Drosophila Medium</td>
		</tr>
		<tr>
			<td>Dental cotton roll (1 x 3.8 cm)</td>
			<td>Gima S.p.A., Gessate, MI, Italy</td>
			<td>35000</td>
			<td>Used for providing water to the parasitoids within the storage container</td>
		</tr>
		<tr>
			<td>Drosophila diet</td>
			<td>Frontier Scientific, Newark, DE, USA</td>
			<td>TF1003</td>
			<td>Custom diet used to rear flies</td>
		</tr>
		<tr>
			<td>Drosophila vial narrow, Polystirene (2.5 x 9.5 cm)</td>
			<td>VWR International, LLC., Radnor, PA, US</td>
			<td>75813-160</td>
			<td>Used for providing water to the parasitoids within the cage</td>
		</tr>
		<tr>
			<td>Drosophila vial plugs, Cellulose acetate (2.5 cm)</td>
			<td>VWR International, LLC., Radnor, PA, US</td>
			<td>89168-886</td>
			<td>Used for providing water to the parasitoids within the cage</td>
		</tr>
		<tr>
			<td>Erlenmeyer flask (250 mL)</td>
			<td>Carolina Biological, Burlington, NC, USA</td>
			<td>731029</td>
			<td>Used for rearing flies and parasitoids</td>
		</tr>
		<tr>
			<td>Falcon-style centrifuge tube (50 mL)</td>
			<td>VWR International, LLC., Radnor, PA, US</td>
			<td>VWRI525-0611</td>
			<td>Modified to ship adult parasitoids</td>
		</tr>
		<tr>
			<td>Foam stopper</td>
			<td>Jaece Industries, North Tanawanda, NY, USA</td>
			<td>L800-C</td>
			<td>Used for sealing the flasks while allowing ventilation for insects</td>
		</tr>
		<tr>
			<td>Honey</td>
			<td>Grocery Store, Newark, DE, USA</td>
			<td>None</td>
			<td>Provided as food for parasitoids</td>
		</tr>
		<tr>
			<td>Identi-Plug plastic foam stopper</td>
			<td>Fisher Scientific Company, L.L.C., Pittsburg, PA, US</td>
			<td>14-127-40E</td>
			<td>Used as feeder for parasitoids and to seal the storage container</td>
		</tr>
		<tr>
			<td>Industrial paper towel</td>
			<td>Grocery Store, Newark, DE, USA</td>
			<td>None</td>
			<td>Provided as a pupation substrate for pupae and mitigated moisture</td>
		</tr>
		<tr>
			<td>Micron mesh fabric (250 mL)</td>
			<td>Industrial Netting, Maple Grove, MN, USA</td>
			<td>WN0250-72</td>
			<td>Used to make ventilation lid for insects</td>
		</tr>
		<tr>
			<td>Nutritional yeast (flakes)</td>
			<td>Grocery Store, Trento, TN, Italy</td>
			<td>None</td>
			<td>Used to prepare the Standard Drosophila Medium</td>
		</tr>
		<tr>
			<td>Paper coaster (10.2 cm)</td>
			<td>Hoffmaster, WI, USA</td>
			<td>35NG26</td>
			<td>Porvided as pupation substrate for flies and parsitized pupae</td>
		</tr>
		<tr>
			<td>Plastic cup (&#216; 13.3 cm, 800 mL)</td>
			<td>Berry Superfos, Taastrup, Denmark</td>
			<td>Unipak 5134</td>
			<td>Modified to store adult parasitoids</td>
		</tr>
		<tr>
			<td>Plastic lid (&#216; 13.3 cm)</td>
			<td>Berry Superfos, Taastrup, Denmark</td>
			<td>PP 2830</td>
			<td>Modified to store adult parasitoids</td>
		</tr>
		<tr>
			<td>Propionic acid</td>
			<td>Merk Life Science S.r.l., Milan, Italy</td>
			<td>P1386 - 1L</td>
			<td>Used to prepare the Standard Drosophila Medium</td>
		</tr>
		<tr>
			<td>Saccharose</td>
			<td>Grocery Store, Trento, TN, Italy</td>
			<td>None</td>
			<td>Used to prepare the Standard Drosophila Medium</td>
		</tr>
		<tr>
			<td>Soup cup with lid (475 mL)</td>
			<td>StackMan, Vietnam</td>
			<td>DC1648</td>
			<td>Used for parasitized larvae to pupate</td>
		</tr>
		<tr>
			<td>Soybean flour</td>
			<td>Grocery Store, Trento, TN, Italy</td>
			<td>None</td>
			<td>Used to prepare the Standard Drosophila Medium</td>
		</tr>
		<tr>
			<td>White felt washer (0.64 cm thick, 5 mm ID x 20 mm OD)</td>
			<td>Quiklok, Lincoln, NH, US</td>
			<td>WFW/.25 x 5 x 20 mm</td>
			<td>Used as feeding ring for parasitoids</td>
		</tr>
	</tbody>
</table>

methods for rearing the parasitoid ganaspis brasiliensis, a promising biological control agent for the invasive drosophila suzukii

斑点翅果蝇原产于东亚，松村 果蝇 （Matsumura）（双翅目：果蝇科）在过去十年中在美洲，欧洲和非洲部分地区广泛建立，成为入侵地区各种软皮水果的毁灭性害虫。生物控制，特别是通过自我延续和特化的寄生虫进行控制，预计将成为对这种高度移动性和多食性害虫进行可持续全区域管理的可行选择。 巴西甘纳斯皮斯 Ihering（膜翅目：Figitidae）是一种广泛分布在东亚的幼虫寄生虫，已被发现是 铃木杜鹃最有效的寄生虫之一。
在对其功效和潜在的非靶向风险进行严格的引入前评估之后，该物种的一个更具有宿主特异性的遗传群体（G1 G. Brasiliensis）最近已被批准在美国和意大利引入和田间释放。另一个遗传群体（G3 G. Brasiliensis）在东亚也很常见，也经常被发现攻击 铃木猪笼草 ，可能会考虑在不久的将来引入。目前，人们对饲养 巴西猪笼草 进行研究或大规模生产以针对 铃木猪笼草的田间释放有着浓厚的兴趣。该协议和相关视频文章描述了这种寄生虫的有效饲养方法，既可以小规模地用于研究，也可以大规模地用于大规模生产和现场释放。这些方法可能有利于进一步的长期研究和使用这种亚洲本地寄生虫作为这种全球入侵害虫的有前途的生物控制剂。

图4 显示了在美国农业部-阿根廷 红新月会益虫 引进处（特拉华州纽瓦克）检疫设施中使用两种不同的寄生虫密度（6对或10对）和两种不同的暴露时间（5或10天）对巴西G3 Ganaspis进行小规模实验室饲养的代表性结果。寄生虫密度和暴露时间的每种组合有14次重复。总共有64个烧瓶生产了4，018只黄蜂（每瓶71.7±4.9个后代），其中49.5%±1.9%的雌性后代。在21°C时，成虫寄生虫...

Watch this Scientific Journal Video about Methods for Rearing the Parasitoid Ganaspis brasiliensis, a Promising Biological Control Agent for the Invasive Drosophila suzukii at JoVE.com

Methods for Rearing the Parasitoid Ganaspis brasiliensis, a Promising Biological Control Agent for the Invasive Drosophila suzukii

巴西果蝇是铃木果蝇（一种全球入侵性水果作物害虫）的幼虫寄生虫，已被批准或考虑引入欧洲和美国，用于生物控制该害虫。本文提供了这种寄生虫的小规模和大规模饲养方案。

巴西寄生体加纳 斯菌的饲养方法，这是一种有前途的生物控制剂，用于侵入性 铃木果蝇

Native to East Asia, the spotted-wing drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), has established widely in the Americas, Europe, ...

生物控制剂的长期研究和使用取决于有效和经济的饲养技术的可用性，并且该协议已被证明是有效的饲养这种寄生虫。目前，人们对饲养这种寄生虫进行研究或现场释放有着巨大的兴趣，该协议将有利于未来长期研究和使用这种有前途的生物控制剂。首先，用户可能需要通过阅读文章中提供的相关参考文献和方法的一些实践来了解所涉及的昆虫的一些基本生物学。为了制作宿主饮食，首先，按照手稿中所述准备饮食的液体部分。接下来，将25至30克捣碎的蓝莓加入空的250毫升烧瓶中。然后，将液体倒入烧瓶中，液体刚好覆盖在捣碎的蓝莓的顶部。接下来，为了饲养宿主铃木果蝇，首先切一块吸水性纸巾，并在中心扭曲它。然后，将纸巾的扭曲中间部分放入饮食瓶中。为了保持水分，使用蒸馏水弄湿纸巾和饮食。然后，为了转移性成熟的成年苍蝇，小心地取出苍蝇菌落瓶的塞子并将其倒置到饮食瓶中。在这种暴露一周后，将苍蝇放在环境室中三周，以便苍蝇出现。为了使宿主幼虫暴露于寄生虫，首先从烧瓶中取出成蝇和纸巾，留下苍蝇卵和幼虫。为了作为幼虫的化蛹基质，折叠一张新的吸水纸巾并将其插入烧瓶中。然后将六对雌性和雄性对的寄生体G3巴西甘纳斯皮斯吸入烧瓶中，并在泡沫塞的底部划出一层薄薄的蜂蜜。为了收集和储存寄生虫，请在五天后取出成虫。然后在前面提到的条件下将烧瓶存放在环境室中35天。随着成年黄蜂的出现，每周吸三次。为了容纳黄蜂，首先将一张湿润但未饱和蒸馏水的纸巾放在2.5×9.5平方厘米果蝇小瓶的底部。然后，将大约60只黄蜂转移到小瓶中并标记出苗日期。将小瓶存放在环境室中。每周两次，在小瓶的泡沫塞底部划出一层薄薄的蜂蜜。首先，安排对果蝇的大规模饲养，将1， 500至2， 000只性成熟的成年苍蝇以50：50的性别比例放在一个大的，编织的，网状的笼子里。然后，将装有水浸棉的盘子和四到六个带有标准果蝇培养基的培养皿放在笼子中。每周两次，用新鲜的培养皿更换暴露的培养皿。将每个没有盖子的受感染培养皿放入800毫升的塑料杯中，并用细网覆盖杯子。然后，将杯子在23摄氏度和75%相对湿度下孵育12至15天。孵化后，将新孵化的成年果蝇铃木转移到饲养笼中。接下来，通过在冷水中冲洗蓝莓一分钟开始准备宿主幼虫，并将水果浸泡在装有漂白剂溶液的盆中，稀释至5%三分钟。然后，用洗过的蓝莓填充九厘米的培养皿。在傍晚时分，将培养皿放入饲养笼中，并放置过夜。第二天早上，轻轻敲击培养皿，将苍蝇从水果中移出，并将水果从饲养笼中取出。同时，为寄生G1巴西加纳斯皮斯准备寄生虫笼和黄蜂出苗笼。两个笼子在天花板下面都应该有一根细绳子，用于悬挂一个或多个喂料器。在每个笼子中，悬挂一个装有水的果蝇小瓶，并根据相对湿度，每五到七天用醋酸纤维素塞从天花板上倒置密封。从铃木果蝇笼中取出受感染的蓝莓后，立即将它们与1， 500至2， 000只巴西加纳斯皮黄蜂一起放入寄生笼中两到三天。接下来，将水果收集在800毫升的塑料杯中，底部有吸水纸。将打开的杯子在21摄氏度和65%相对湿度下在出苗笼或外爆笼中孵育至少28天。在孵化的第二周和第三周，每周检查笼子是否有早期宿主外闭。为了便于连续的寄生虫收集，切除成蝇。在孵化的第四周结束时，为笼子提供喂食器和水源。收集约10%至15%的成年黄蜂，并将其转移到寄生笼中，以取代旧的和无生产力的黄蜂。从外阴笼的剩余黄蜂中，以50：50的性别比例储存多达700只成虫，其中800毫升塑料杯包含一个装满水的两毫升管，并在杯子底部用牙科棉卷密封。将具有明显出苗状态的杯子存放在环境室中长达一个月。对于运输寄生虫，将约200个成年人放在50毫升的锥形管中。然后，将所有管子与冰袋一起放入绝缘运输容器中。该协议导致在实验室中成功地小规模饲养了G3巴西加纳斯皮斯，一种寄生虫剂，针对农业害虫铃木果蝇。后代生产不受寄生虫密度和暴露时间差异的影响。G1巴西猪笼草的大规模生成也可以实现，在最初暴露于宿主铃木果蝇后五周开始生产。从第8周到第22周，寄生虫的产生与暴露于它们的水果量成正比。保持合适的饲养温度，控制湿度并始终为成年昆虫提供食物至关重要。通过优化寄主和寄生体的饲养密度以最大限度地提高黄蜂的繁殖，并探索在替代寄主上饲养这种寄生虫的可能性，可以进一步改进这些方法。

Research

JoVE Journal

Biology

Methods for Rearing the Parasitoid Ganaspis brasiliensis, a Promising Biological Control Agent for the Invasive Drosophila suzukii

Protocol for Mosquito Rearing (A. gambiae)

Protocol for Mosquito Rearing A. gambiae

蚊子养育的议定书“（冈比亚按蚊）

This protocol describes mosquito rearing in the insectary. The insectary rooms are maintained at 28&deg;C and ~80% humidity, with a 12 hr. day/night ...

科研

生物学

Labeling hESCs and hMSCs with Iron Oxide Nanoparticles for Non-Invasive in vivo Tracking with MR Imaging

氧化铁标记的胚胎干细胞和的hMSCs纳米粒子在体内的跟踪与非侵入性磁共振成像

In recent years, stem cell research has led to a better understanding of developmental biology, various diseases and its potential impact on regenerative ...

Labeling Stem Cells with Fluorescent Dyes for non-invasive Detection with Optical Imaging

与非侵入性检测荧光染料标记的干细胞与光学成像

Optical imaging (OI) is an easy, fast and inexpensive tool for in vivo monitoring of new stem cell based therapies. The technique is based on ex vivo ...

Electrophysiological Methods for Recording Synaptic Potentials from the NMJ of Drosophila Larvae

电生理方法记录从果蝇幼虫NMJ突触潜力

In this video, we describe the electrophysiological methods for recording synaptic transmission at the neuromuscular junction (NMJ) of Drosophila larva. ...

Electrophysiological Recording in the Drosophila Embryo

Electrophysiological Recording in the Drosophila Embryo

在电生理记录果蝇胚胎

Drosophila is a premier genetic model for the study of both embryonic development and functional neuroscience. Traditionally, these fields are quite ...

Visualizing the Beating Heart in Drosophila

Visualizing the Beating Heart in Drosophila

可视化心脏跳动果蝇</em

The Drosophila heart has recently emerged as a good model system for examining the genetic, cellular, and molecular mechanisms underlying function in ...

Methods for the Study of the Zebrafish Maxillary Barbel

上颌Barbel斑马鱼的研究方法

Barbels are skin sensory appendages found in fishes, reptiles and amphibians.  The zebrafish, Danio rerio, develops two pairs of barbels- a short nasal ...

An Optimized Protocol for Rearing Fopius arisanus, a Parasitoid of Tephritid Fruit Flies

An Optimized Protocol for Rearing Fopius arisanus, a Parasitoid of Tephritid Fruit Flies

一个优化的协议饲养 Fopius arisanus Tephritid果寄生蝇

Fopius arisanus (Sonan) is an important parasitoid of Tephritid fruit flies for at least two reasons. First, it is the one of only three opiine ...

Blastomere Explants to Test for Cell Fate Commitment During Embryonic Development

卵裂球外植体测试胚胎发育过程中细胞命运的承诺

Fate maps, constructed from lineage tracing all of the cells of an embryo, reveal  which tissues descend from each cell of the embryo.  Although fate maps ...

Autonomously Bioluminescent Mammalian Cells for Continuous and Real-time Monitoring of Cytotoxicity

自主的生物发光哺乳动物细胞连续和实时监测细胞毒性

Mammalian  cell-based in vitro assays have been  widely employed as alternatives to animal testing for toxicological studies but  have been limited due to ...

巴西寄生体加纳 斯菌的饲养方法，这是一种有前途的生物控制剂，用于侵入性 铃木果蝇

巴西寄生体加纳斯菌的饲养方法，这是一种有前途的生物控制剂，用于侵入性铃木果蝇