我们感谢基斯Shigeteni insectary和娜塔莎Sostrom援助帮助与计算机图形。这项工作是由美国农业部资助。本出版物中所表达的意见，研究成果，结论或建议是作者的，不一定反映美国农业部的意见。美国农业部是一个机会均等的提供者和雇主。

1。准备主机果蝇卵寄生<ol><li>准备准备琼脂（另外，Gelcarin GP812，FMC生物聚合物，尤因新泽西州，更具成本效益的选项）寄生的果蝇卵充满基板的菜肴，每边长10厘米和1.5厘米的深度。准备在9克每公升水的浓度的琼脂。 </li><li>填写菜肴与液体培养基（约70毫升）的边缘，并让他们冷却和固化（至少45分钟）。 </li><li>一旦固体琼脂块，应用纸巾单层（单层的乔治亚太平洋"偏爱"批次组织），覆盖了每个块的顶部。组织应坚持容易因表面水分的琼脂。 </li><li>套用一个0.5毫升的量果蝇鸡蛋悬浮在水中的组织覆盖的琼脂块表面。在我们的insectary，这相当于约6000桔小实蝇（每毫升12000蛋）鸡蛋。 </li><li>使用干净的2.5厘米宽刷蘸水果实蝇卵均匀地分布在组织表面。 </li><li>将果蝇卵蜂笼的底部屏幕下允许产卵。点击屏幕底部的底面，以去除死的寄生虫，这可能会阻碍鸡蛋。这是最好的屏幕底部，如果不碰蛋。 </li><li>允许寄生虫排卵过夜，到第二天，约21小时。这是高比例的寄生蛋足够的时间。较长的曝光风险superparasitization。 </li></ol> 2。孵化和化蛹寄生主机果实蝇<ol><li>准备饲养容器与朝上鸡蛋放置3至3.5每1.25升果蝇饮食19的寄生蛋的准备琼脂块。 </li><li>饮食容器放置在超过一个1.5厘米的细级蛭石或冲砂深层化蛹容器在一个较大的的提升的蛋。化蛹容器盖上盖子和胶带与胶带边缘。蛭石可重复使用过筛后（见下文）。 </li><li>确保有足够的空间之间的饮食容器的顶部和化蛹容器的盖子，果蝇幼虫爬出来，"流行"到下面 vemiculite 20。 </li><li>化蛹容器移动到一个房间在27 ° C和80％相对湿度（RH）为一个星期。与前四天暗布盖的容器，然后取出余下的日子里布。 </li><li>一周后取出幼虫的饮食，一旦幼虫弹出，进入化蛹蛭石。如果有幼虫在蛭石，让他们一夜之间化蛹。 </li><li>筛从蛭石蛹。开始用粗筛（约3毫米），然后用一只手筛，蛭石中删除任何剩余的团块。 </li></ol> 3。排序和选择<ol><li>蚕蛹含有F. arisanus可以部分隔离unparasitized果蝇大小。收集蛹1.65和2.26毫米之间，直径，因为这些将丰富寄生蝇 12 Puparia较小，大多包含矮小的寄生蜂，更大的蛹大多含有苍蝇。还要注意，寄生蛹的女性比例呈正相关， 大小为12 。 </li><li>可能采用多种方法，按大小排序的蛹。我们将描述一个机械的方法，使用自定义大小分拣机 21 。 </li><li>放置在一个振动给料机慢慢饲料大小排序顶部的漏斗中的蛹。 </li><li>蛹送入大小分拣机，其中包括一对在30 °的角度稍有分歧，滚动酒吧。单独成一个插槽，分为十个不同的容器通过一个漏斗空阵列大小蛹下降。 </li><li>从尺寸范围在1.65至2.26毫米的杯子收集蛹和他们出现在一个笼子里放置7天以上，直到大多数苍蝇unparasitized蛹出现。 </li><li>接下来，使用一个风扇分开空puparia那些仍然含有昆虫。 </li></ol> 4 楼arisanus出现和维护<ol><li>为了保持F 的殖民地arisanus使用一个可移动的玻璃前一个寄生的控股笼，约25厘米，每边长。笼子的顶部和两侧两个1毫米2甄别。笼的后面是在为9厘米的洞访问的橡胶板覆盖。笼子的底部为1 - 2mm 2的屏幕大到足以让两个琼脂块的位置与果蝇卵寄生蜂产卵在里面长满了削减。 </li><li>控股笼也应该有一个沿着前面的玻璃底部季度的有色部分，以尽量减少光可以创建沿笼前蜂拥挤，导致死亡率升高。 </li><li>将这些选择性约11克特德蛹成塑料容器直径约9厘米。这应该产生约600-700每笼寄生虫。尺寸的选择应导致约60％为女性。 </li><li>将屏蔽盖的容器。盖子应覆盖2毫米2筛查，这将允许成年楼arsianus离开后出现的容器，但将包含任何剩余的果蝇。 </li><li>寄生虫保持在24℃，相对湿度约45％具有12:12的光照和通风良好。如果死亡率高，户外移动了几个小时，每天或增加通风的笼子，可能会降低。寄生虫应准备排卵，在延长一个星期，应为两个星期的生产。 </li><li>纺沿着笼子的顶部数稀释的蜂蜜至少3次每星期或可用蜂蜜干燥时。应用在窄条纹，用指尖旋转蜂蜜。请注意，如果条纹太重的蜂蜜滴进笼子的蜂蜜，如果太淡会干出快速和需要频繁地重新申请。 </li><li>将琼脂块（10厘米× 10厘米× 4厘米），笼子的顶部提供水分寄生虫。这些应改为每周两次。 </li></ol> 5。代表性的成果: 我们在这里报告美国农业部F.进行质量控制程序的结果arisanus饲养在希洛，夏威夷1月5日和2010年6月22日之间的操作。在希洛的位置就在这个视频文章和协议所描述的规模养殖开始于2009年8月，最初的问题，并与新的位置调整已大多由一月解决。因此，这些数据的结果可能会得到其他研究人员开始他们的殖民地，如果他们按照协议所述的代表，一般有经验的饲养昆虫。在此期间，我们保持了 F的小殖民地arisanus:4寄生控股笼每周产生，相当于约3600 楼arisanus。 丰富的寄生率的初步检查即幼虫的饮食后，立即被删除，并进行的蛹2 g样品在尺寸范围为1.65 - 前一家控股笼发展的一个额外的一周（2.26毫米（丰富）蛹筛选和大小排序，我们称之为"初蛹"这些）。在丰富的样品在2010年首3个月的平均比例寄生虫为0.46（SD = 0.18）。这意味着在下一季度为0.58（SD = 0.08），反映了在新的位置的殖民地和饲养程序的稳定。请注意，没有成功寄生蛹的丰富，包括那些没有产生emergents产生苍蝇。 在为期一周的额外空puparia控股笼拆除后另两个克样品被送往（"晚蛹"）。果蝇代表根据这第二个样本的1％。果蝇的比例很低，部分原因是较早出现苍蝇相比，寄生虫，所以在控股容器保持最苍蝇。 最后，图1给出了从1月至2010年6月的产量在蛹质量。产量是衡量从蛹利益的大小范围，丰富了1.65-2.26毫米。 "初蛹"在今年年初的大肿块，表明果蝇的高比例和unemerged当时蛹，而正在作出调整，新的设施。 <img src="/files/ftp_upload/2901/2901fig1.jpg" alt="图1" /> 图1。产量丰富（蛹直径1.652.26毫米），早期（大小选择后立即）和"晚"（后一个星期，删除空弹壳）的ARS，USDA F. ariasnus殖民地蛹在希洛，夏威夷2010年1月 - 6月

<ol>
	<li>Wang, X. G., Bokonon-Ganta, A. H., Ramadan, M. M., Messing, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Egg-larval+opiine+parasitoids+(Hym.,+Braconidae)+of+tephritid+fruit+fly+pests+do+not+attack+the+flowerhead-feeder+Trupanea+dubautiae+(Dipt.,+Tephritidae.">Egg-larval opiine parasitoids (Hym., Braconidae) of tephritid fruit fly pests do not attack the flowerhead-feeder Trupanea dubautiae (Dipt., Tephritidae.</a> Journal of Applied Entomology. 128, 716-722 (2004).</li><li>Vargas, R. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potential+for+areawide+integrated+management+of+Mediterranean+fruit+fly+(Diptera+:+Tephritidae)+with+a+braconid+parasitoid+and+a+novel+bait+spray.">Potential for areawide integrated management of Mediterranean fruit fly (Diptera : Tephritidae) with a braconid parasitoid and a novel bait spray.</a> Journal of Economic Entomology. 94, 817-825 (2001).</li><li>Vargas, R. I., Leblanc, L., Putoa, R., Eitam, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+introduction+of+Bactrocera+dorsalis+(Diptera+:+Tephritidae)+and+classical+biological+control+releases+of+Fopius+arisanus+(Hymenoptera+:+Braconidae)+on+economically+important+fruit+flies+in+French+Polynesia.">Impact of introduction of Bactrocera dorsalis (Diptera : Tephritidae) and classical biological control releases of Fopius arisanus (Hymenoptera : Braconidae) on economically important fruit flies in French Polynesia.</a> Journal of Economic Entomology. 100, 670-679 (2007).</li><li>Harris, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suppression+of+melon+fly+(Diptera:+Tephritidae)+populations+with+releases+of+Fopius+arisanus+and+Psyttalia+fletcheri+(Hymenoptera+Braconidae)+in+North+Shore+Oahu,+HI,+USA.">Suppression of melon fly (Diptera: Tephritidae) populations with releases of Fopius arisanus and Psyttalia fletcheri (Hymenoptera Braconidae) in North Shore Oahu, HI, USA.</a> Biocontrol. 55, 593-599 (2010).</li><li>Haramoto, F. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Biology of Opius oophilus Fullaway (Hymenoptera: Braconidae) [dissertation]. , (1988).</li><li>Chong, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Production+methods+for+fruit+fly+parasites.">Production methods for fruit fly parasites.</a> Proceedings of the Hawaiian Entomological Society. 18, 61-63 (1962).</li><li>Snowball, G. J., Wilson, F., Lukins, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Culture+and+consignment+techiques+used+for+parasites+introduced+against+Queensland+fruit+fly+(Strumeta+tryoni+(Frogg).).">Culture and consignment techiques used for parasites introduced against Queensland fruit fly (Strumeta tryoni (Frogg).).</a> Australian Journal of Agricultural Research. 13, 233-248 (1962).</li><li>Ramadan, M. M., Wong, T. T. Y., Beardsley, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reproductive+Behavior+of+Biosteres+arisanus+(Sonan)+(Hymenoptera:Braconidae),+an+Egg-Larval+Parasitoid+of+the+Oriental+Fruit+Fly.">Reproductive Behavior of Biosteres arisanus (Sonan) (Hymenoptera:Braconidae), an Egg-Larval Parasitoid of the Oriental Fruit Fly.</a> Biological Control. 2, 28-34 (1992).</li><li>Ramadan, M. M., Wong, T. T. Y., McInnis, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reproductive+biology+of+Biosteres+arisanus+(Sonan),+an+egg-larval+parasitoid+of+the+oriental+fruit+fly.">Reproductive biology of Biosteres arisanus (Sonan), an egg-larval parasitoid of the oriental fruit fly.</a> Biological Control. 4, 93-100 (1994).</li><li>Harris, E. J., Okamoto, R. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Method+for+rearing+Biosteres+arisanus+(Hymenoptera,+Braconidae)+in+the+laboratory.">A Method for rearing Biosteres arisanus (Hymenoptera, Braconidae) in the laboratory.</a> Journal of Economic Entomology. 84, 417-422 (1991).</li><li>Bautista, R. C., Harris, E. J., Lawrence, P. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biology+and+rearing+of+the+fruit+fly+parasitoid+Biosteres+arisanus:+clues+to+insectary+propagation.">Biology and rearing of the fruit fly parasitoid Biosteres arisanus: clues to insectary propagation.</a> Entomologia Experimentalis Et Applicata. 89, 79-85 (1998).</li><li>Bautista, R. C., Mochizuki, N., Spencer, J. P., Harris, E. J., Ichimura, 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=Mass-rearing+of+the+tephritid+fruit+fly+parasitoid+Fopius+arisanus+(Hymenoptera+:+Braconidae).">Mass-rearing of the tephritid fruit fly parasitoid Fopius arisanus (Hymenoptera : Braconidae).</a> Biological Control. 15, 137-144 (1999).</li><li>Bautista, R. C., Harris, E. J., Vargas, R. I., Jang, E. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parasitization+of+melon+fly+(Diptera+:+Tephritidae)+by+Fopius+arisanus+and+Psyttalia+fletcheri+(Hymenoptera+:+Braconidae)+and+the+effect+of+fruit+substrates+on+host+preference+by+parasitoids.">Parasitization of melon fly (Diptera : Tephritidae) by Fopius arisanus and Psyttalia fletcheri (Hymenoptera : Braconidae) and the effect of fruit substrates on host preference by parasitoids.</a> Biological Control. 30, 156-164 (2004).</li><li>Rousse, P., Gourdon, F., Quilici, S. <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+the+egg+pupal+parasitoid+Fopius+arisanus+(Hymenoptera+:+Braconidae)+in+La+Reunion.">Host specificity of the egg pupal parasitoid Fopius arisanus (Hymenoptera : Braconidae) in La Reunion.</a> Biological Control. 37, 284-290 (2006).</li><li>Quimio, G. M., Walter, G. H. <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+and+host+suitability+in+an+egg-pupal+fruit+fly+parasitoid,+Fopius+arisanus+(Sonan)+(Hym.,+Braconidae).">Host preference and host suitability in an egg-pupal fruit fly parasitoid, Fopius arisanus (Sonan) (Hym., Braconidae).</a> Zeitschrift fur Angewandtes Entomologie. 125, 135-140 (2001).</li><li>Calvitti, M., Antonelli, M., Moretti, R., Bautista, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oviposition+response+and+development+of+the+egg-pupal+parasitoid+Fopius+arisanus+on+Bactrocera+oleae,+a+tephritid+fruit+fly+pest+of+olive+in+the+Mediterranean+basin.">Oviposition response and development of the egg-pupal parasitoid Fopius arisanus on Bactrocera oleae, a tephritid fruit fly pest of olive in the Mediterranean basin.</a> Entomologia Experimentalis Et Applicata. 102, 65-73 (2002).</li><li>Montoya, P., Suarez, A., Lopez, F., Cancino, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fopius+arisanus+oviposition+in+four+Anastrepha+fruit+fly+species+of+economic+importance+in+Mexico.">Fopius arisanus oviposition in four Anastrepha fruit fly species of economic importance in Mexico.</a> Biocontrol. 54, 437-444 (2009).</li><li>Vargas, R. I., Shelly, T. E., Leblanc, L., Pi&#241;ero, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+Advances+in+Methyl+Eugenol+and+Cue-Lure+Technologies+for+Fruit+Fly+Detection,+Monitoring,+and+Control+in+Hawaii.">Recent Advances in Methyl Eugenol and Cue-Lure Technologies for Fruit Fly Detection, Monitoring, and Control in Hawaii.</a> Pheromones. 83, 575-595 (2010).</li><li>Tanaka, N., Steiner, L. F., Ohinata, K., Okamoto, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low-cost+larval+rearing+medium+for+mass+production+of+oriental+and+Mediterranean+fruit+flies.">Low-cost larval rearing medium for mass production of oriental and Mediterranean fruit flies.</a> Journal of Economic Entomology. 62, 967-968 (1969).</li><li>Vargas, R. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mass+production+of+tephritid+fruit+flies.">Mass production of tephritid fruit flies.</a> World crop pests. Fruit flies: Their biology, natural enemies and control. 3, 141-151 (1989).</li><li>Spencer, J. P., Mochizuki, N., McInnis, D. O., Liquido, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanical+separation+of+parasitoid+sexes+based+upon+size+of+fruit+fly+host+pupae.">Mechanical separation of parasitoid sexes based upon size of fruit fly host pupae.</a> , (1996).</li><li>Dukas, R., Duan, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potential+fitness+consequences+of+associative+learning+in+a+parasitoid+wasp.">Potential fitness consequences of associative learning in a parasitoid wasp.</a> Behavioral Ecology. 11, 536-543 (2000).</li><li>Drew, R., Tsuruta, K., White, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+species+of+pest+fruit+fly+(Diptera:+Tephritidae:+Dacinae)+from+Sri+Lanka+and+Africa.">A new species of pest fruit fly (Diptera: Tephritidae: Dacinae) from Sri Lanka and Africa.</a> African Entomology. 13, 149-154 (2005).</li><li>Argov, Y., Gazit, Y. <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+the+Mediterranean+fruit+fly+in+Israel:+Introduction+and+establishment+of+natural+enemies.">Biological control of the Mediterranean fruit fly in Israel: Introduction and establishment of natural enemies.</a> Biological Control. 46, 502-507 (2008).</li><li>Bokonon-Ganta, A. H., Ramadan, M. M., Messing, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reproductive+biology+of+Fopius+ceratitivorus+(Hymenoptera+:+Braconidae),+an+egg-larval+parasitold+of+the+Mediterranean+fruit+fly,+Ceratitis+capitata+(Diptera+:+Tephritidae).">Reproductive biology of Fopius ceratitivorus (Hymenoptera : Braconidae), an egg-larval parasitold of the Mediterranean fruit fly, Ceratitis capitata (Diptera : Tephritidae).</a> Biological Control. 41, 361-367 (2007).</li></ol>

在本议定书所附的视频文章，我们已经描述并演示了一个优化的协议 ，饲养楼ariasnus，Tephritid果蝇寄生在实验室环境中， 。多年来，该协议已经细化，以尽量减少所需的劳动力和专门设备，以维持一个殖民地。我们注意到，需要一个完善的，生产和稳定主机果蝇的殖民地饲养寄生蜂的任何企图。我们的insectary，我们在使用一台主机果蝇桔小实蝇，但其他物种已被证明才能胜任的主机 ，以及14-17。 F.几个方面arisanus殖民地维护有待探讨。这些措施包括这 10种在实验室适应率， 学习 22的机制和遗传变化与适应，这可能是在主机果蝇可塑性的机制，可能会发生。 在过去的十多年，已有Dacine苍蝇传播世界各地的多个例子，尤其是那些属实蝇:在法属波利尼西亚，B.的桔小实蝇在南美洲，B部分carambolae在非洲和 B invadens zonata在非洲和地中海北部如​​F.生物控制剂的有效性。3,23,18测试实蝇对其他物种的 arisanus应予以高度重视，这是我们希望在本议定书和影随行的文章中介绍的方法，应用程序将加速F上的研究arisanus在更广泛的地点24。最后，进一步研究为出发点，使用这种方法也可提供重要的信息，以新颖的卵寄生蜂的定植 25.1

an optimized protocol for rearing fopius arisanus, a parasitoid of tephritid fruit flies

Fopius arisanus（Sonan）是一个重要的寄生的Tephritid果蝇，至少有两个原因。首先，它是一个只有三个opiine寄生蜂，已知感染的主机在卵期 1 。其次，它具有广泛的潜在果蝇主机。也许是由于它的生活历史 ，F. arisanus已成功用于控制在2-4多个热带地区的生物果蝇。 F.广泛使用的障碍之一控制果蝇的 arisanus是，它是难以建立一个稳定的实验室殖民地 5-9 。尽管这一困难，在20世纪90年代，美国农业部的研究人员开发了一种可靠的方法来维持楼的实验室种群arisanus 10-12。有极大的兴趣，是在 F arisanus生物学13,14，尤其是关于其能力殖民各种各样的Tephritid主机14-17;利益尤其是惊人的桔小实蝇害虫蔓延驱动，在过去十年中的新大洲18。 F 上的进一步研究arisanus和额外部署这一物种作为生物控制剂，将有利于从饲养方式的优化和改进。在这个协议和相关视频的文章中，我们描述了一个优化的方法饲养楼arisanus基于先前描述的方法 12 。我们在这里描述的方法，使饲养楼在没有使用的水果的小规模 arisanus，使用的材料在世界各地的热带地区与相对较低的体力劳动要求。

Watch this Scientific Journal Video about An Optimized Protocol for Rearing Fopius arisanus, a Parasitoid of Tephritid Fruit Flies at JoVE.com

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

Fopius arisanus Tephritid水果，是成功地使用这些重要的热带害虫的生物控制苍蝇的卵幼虫寄生。我们在这里描述饲养楼的优化协议arisanus在小规模使用现成材料。

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 ...

an-optimized-protocol-for-rearing-fopius-arisanus-parasitoid

Research

JoVE Journal

Biology

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

14.7K 次观看.  US Pacific Basin Agricultural Research Center. Fopius arisanus Tephritid水果，是成功地使用这些重要的热带害虫的生物控制苍蝇的卵幼虫寄生。我们在这里描述饲养楼的优化协议arisanus在小规模使用现成材料。

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

Studying Aggression in Drosophila (fruit flies)

Aggression is an innate behavior that evolved in the framework of defending or obtaining resources. This complex social behavior is influenced by genetic, hormonal and environmental factors. In many organisms, aggression is critical to survival but controlling and suppressing aggression in distinct contexts also has become increasingly important. In recent years, invertebrates have become increasingly useful as model systems for investigating the genetic and systems biological basis of complex social behavior. This is in part due to the diverse repertoire of behaviors exhibited by these organisms. In the accompanying video, we outline a method for analyzing aggression in Drosophila whose design encompasses important eco-ethological constraints. Details include steps for: making a fighting chamber; isolating and painting flies; adding flies to the fight chamber; and video taping fights. This approach is currently being used to identify candidate genes important in aggression and in elaborating the neuronal circuitry that underlies the output of aggression and other social behaviors.

Studying Aggression in Drosophila fruit flies

学习在果蝇的侵略（果蝇）

Aggression is an innate behavior that evolved in the framework of defending or obtaining resources.  This complex social behavior is influenced by ...

科研

生物学

Protocol for Mosquito Rearing (A. gambiae)

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 cycle. For this procedure, you'll need mosquito cages, 10% sterile sucrose solution, paper towels, beaker, whatman filter paper, glass feeders, human blood and serum, water bath, parafilm, distilled water, clean plastic trays, mosquito food (described below), mosquito net to cover the trays, vacuum, and a collection chamber to collect adults.

Protocol for Mosquito Rearing A. gambiae

This video illustrates the general techniques used to rear Anopheles gambiae in the laboratory. The methods for caring for laboratory mosquitoes are demonstrated through all stages of the organism's life cycle from larvae to pupae to blood-feeding adults.

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 ...

A Protocol for the Production of KLRG1 Tetramer

Killer cell lectin-like receptor G1 (KLRG1) is a type II transmembrane glycoprotein inhibitory receptor belonging to the C type lectin-like superfamily. KLRG1 exists both as a monomer and as a disulfide-linked homodimer. This well-conserved receptor is found on the most mature and recently activated NK cells as well as on a subset of effector/memory T cells.
Using KLRG1 tetramer as well as other methods, E-, N-, and R-cadherins were identified as KLRG1 ligands. These Ca2+-dependent cell-cell adhesion molecules comprises of an extracellular domain containing five cadherin repeats responsible for cell-cell interactions, a transmembrane domain and a cytoplasmic domain that is linked to the actin cytoskeleton. 
Generation of the KLRG1 tetramer was essential to the identification of the KLRG1 ligands. KLRG1 tetramer is also a unique tool to elucidate the roles cadherin and KLRG1 play in regulating the immune response and tissue integrity.

This protocol describes the production of KLRG1 tetramer, which is a powerful tool for the analysis of KLRG1 ligands.

KLRG1四聚体的生产协议

Killer cell lectin-like receptor G1 (KLRG1) is a type II transmembrane glycoprotein inhibitory receptor belonging to the C type lectin-like superfamily. ...

Isolation and Biophysical Study of Fruit Cuticles

The cuticle, a hydrophobic protective layer on the aerial parts of terrestrial plants, functions as a versatile defensive barrier to various biotic and abiotic stresses and also regulates water flow from the external environment.1 A biopolyester (cutin) and long-chain fatty acids (waxes) form the principal structural framework of the cuticle; the functional integrity of the cuticular layer depends on the outer 'epicuticular' layer as well as the blend consisting of the cutin biopolymer and 'intracuticular' waxes.2 Herein, we describe a comprehensive protocol to extract waxes exhaustively from commercial tomato (Solanum lycopersicum) fruit cuticles or to remove epicuticular and intracuticular waxes sequentially and selectively from the cuticle composite. The method of Jetter and Sch&auml;ffer (2001) was adapted for the stepwise extraction of epicuticular and intracuticular waxes from the fruit cuticle.3,4 To monitor the process of sequential wax removal, solid-state cross-polarization magic-angle-spinning (CPMAS) 13C NMR spectroscopy was used in parallel with atomic force microscopy (AFM), providing molecular-level structural profiles of the bulk materials complemented by information on the microscale topography and roughness of the cuticular surfaces. To evaluate the cross-linking capabilities of dewaxed cuticles from cultivated wild-type and single-gene mutant tomato fruits, MAS 13C NMR was used to compare the relative proportions of oxygenated aliphatic (CHO and CH2O) chemical moieties.
Exhaustive dewaxing by stepwise Soxhlet extraction with a panel of solvents of varying polarity provides an effective means to isolate wax moieties based on the hydrophobic characteristics of their aliphatic and aromatic constituents, while preserving the chemical structure of the cutin biopolyester. The mechanical extraction of epicuticular waxes and selective removal of intracuticular waxes, when monitored by complementary physical methodologies, provides an unprecedented means to investigate the cuticle assembly: this approach reveals the supramolecular organization and structural integration of various types of waxes, the architecture of the cutin-wax matrix, and the chemical composition of each constituent. In addition, solid-state 13C NMR reveals differences in the relative numbers of CHO and CH2O chemical moieties for wild-type and mutant red ripe tomato fruits. The NMR techniques offer exceptional tools to fingerprint the molecular structure of cuticular materials that are insoluble, amorphous, and chemically heterogeneous. As a noninvasive surface-selective imaging technique, AFM furnishes an effective and direct means to probe the structural organization of the cuticular assembly on the nm-&mu;m length scale.

Aerial plant organs are protected by the cuticle, a supramolecular biopolyester-wax assembly. We present protocols to monitor selective removal of epi- and intracuticular waxes from tomato fruit cuticles on molecular and micro scales by solid-state NMR and atomic force microscopy, respectively, and to assess the cross-linking capacity of engineered cuticular biopolyesters.

果实角质层分离和生物物理研究

The cuticle, a hydrophobic protective layer on the aerial parts of terrestrial plants, functions as a versatile defensive barrier to various biotic and ...

Fruit Volatile Analysis Using an Electronic Nose

Numerous and diverse physiological changes occur during fruit ripening, including the development of a specific volatile blend that characterizes fruit aroma. Maturity at harvest is one of the key factors influencing the flavor quality of fruits and vegetables1. The validation of robust methods that rapidly assess fruit maturity and aroma quality would allow improved management of advanced breeding programs, production practices and postharvest handling. 
Over the last three decades, much research has been conducted to develop so-called electronic noses, which are devices able to rapidly detect odors and flavors2-4. Currently there are several commercially available electronic noses able to perform volatile analysis, based on different technologies. The electronic nose used in our work (zNose, EST, Newbury Park, CA, USA), consists of ultra-fast gas chromatography coupled with a surface acoustic wave sensor (UFGC-SAW). This technology has already been tested for its ability to monitor quality of various commodities, including detection of deterioration in apple5; ripeness and rot evaluation in mango6; aroma profiling of thymus species7; C6 volatile compounds in grape berries8; characterization of vegetable oil9 and detection of adulterants in virgin coconut oil10. 
This system can perform the three major steps of aroma analysis: headspace sampling, separation of volatile compounds, and detection. In about one minute, the output, a chromatogram, is produced and, after a purging cycle, the instrument is ready for further analysis. The results obtained with the zNose can be compared to those of other gas-chromatographic systems by calculation of Kovats Indices (KI). Once the instrument has been tuned with an alkane standard solution, the retention times are automatically converted into KIs. However, slight changes in temperature and flow rate are expected to occur over time, causing retention times to drift. Also, depending on the polarity of the column stationary phase, the reproducibility of KI calculations can vary by several index units11. A series of programs and graphical interfaces were therefore developed to compare calculated KIs among samples in a semi-automated fashion. These programs reduce the time required for chromatogram analysis of large data sets and minimize the potential for misinterpretation of the data when chromatograms are not perfectly aligned.
We present a method for rapid volatile compound analysis in fruit. Sample preparation, data acquisition and handling procedures are also discussed.

A rapid method for volatile compound analysis in fruit is described. The volatile compounds present in the headspace of a homogenate of the sample are rapidly separated and detected with ultra-fast gas chromatography (GC) coupled with a surface acoustic wave (SAW) sensor. A procedure for data handling and analysis is also discussed.

果实挥发分析，利用电子鼻

Numerous and diverse physiological changes occur during fruit ripening, including the development of a specific volatile blend that characterizes fruit ...

Optimized Protocol for Retinal Wholemount Preparation for Imaging and Immunohistochemistry

Working with delicate tissue can be a complicating factor when performing immunohistochemical assessment. Here, we present a method that utilizes a ring-supported hydrophilized PTFE membrane to provide structural support to both living and fixed tissue during immunohistochemical processing, which allows for the use of a variety of protocols that would otherwise cause damage to the tissue. First, this is demonstrated with bolus loading of fluorescent markers into living retinal tissue. This method allows for quick visualization of targeted structures, while the membrane support maintains tissue integrity during the injection and allows for easy transfer of the preparation for further imaging or processing.
Second, a procedure for antibody staining in tissue fixed with carbodiimide is described. Though paraformaldehyde fixation is more common, carbodiimide fixation provides a superior substrate for the visualization of synaptic proteins. A limitation of carbodiimide is that the resulting fixed tissue is relatively fragile; however, this is overcome with the use of the supporting membrane. Retinal tissue is used to demonstrate these techniques, but they may be applied to any fragile tissue.

The protocol described here is for structural assessment of a wholemount retinal preparation. This includes descriptions of tissue dissection, mounting onto a hydrophilized polytetrafluoroethylene (PTFE) membrane insert, bolus loading with fluorescent markers, and a comparison of fixation with carbodiimide and paraformaldehyde for immunohistochemical analysis of cellular and synaptic components.

视网膜平片准备成像和免疫组化优化方案

Working with delicate tissue can be a complicating factor when performing immunohistochemical assessment. Here, we present a method that utilizes a ...

Laboratory Rearing of Stable Flies and Other Muscoid Diptera

Stable flies, Stomoxys calcitrans, are serious pests of livestock, humans, companion animals and wildlife worldwide. During the last 20+ years, changes in agronomic practices resulted in serious outbreaks of stable flies in several countries. These outbreaks disrupted livestock production and human recreation resulting in public demands for increasing research and management efforts for this pest. A simple and inexpensive procedure for rearing stable flies for laboratory studies is presented. The procedure uses locally available diet components, equipment and supplies. The procedure can be adapted for rearing other muscoid flies including face fly (Musca autumnalis), horn fly (Haematobia irritans), and house fly (Musca domestica). The procedure produces stable fly puparia averaging 12.5 mg and ~35% egg to adult survival. Approximately 3000 flies are produced in each pan.

A procedure for rearing stable flies (Stomoxys calcitrans) is presented. The procedure uses locally available materials for diet components, equipment and supplies.

稳定蝇及其他 Muscoid 双翅目的实验室饲养

Stable flies, Stomoxys calcitrans, are serious pests of livestock, humans, companion animals and wildlife worldwide. During the last 20+ years, changes in ...

Simple Homemade Tools to Handle Fruit Flies&#8212;Drosophila melanogaster

The fruit fly, Drosophila melanogaster, is widely used both in biological research and biology education. Handling adult flies is common but difficult in practice, as adult flies fly. Demonstrated here is how to make some simple and cost-effective tools to address difficult issues in the handling of Drosophila. Holes in foam stoppers are made and pipette tips or funnels are inserted into the holes. Flies then move only in one direction into the pipette tip/funnel assemblage, allowing efficient control of the transfer of adult Drosophila into or out of a vial. Existing protocols have been modified for cool-anesthetizing flies by chilling in crushed ice and transferring them onto a cold, hard icepack surface. The icepack is covered with a piece of medical gauze that keeps immobilized flies from the condensed water when examined under a stereomicroscope. The flies are finally euthanized for counting and sorting or discarded by microwaving. A bottle-shaped cage has also been developed for collecting eggs, as well as a labor-saving device and accompanying protocol for cleaning glass culture vials.

Described here is the use of several homemade tools to transfer, chill, and kill adult Drosophila, as well as to clean glass culture vials and collect eggs. These tools are easy to make and are rather efficient in handling Drosophila.

简单的自制工具处理果蝇*果蝇黑色素

The fruit fly, Drosophila melanogaster, is widely used both in biological research and biology education. Handling adult flies is common but difficult in ...

Topical Application Bioassay to Quantify Insecticide Toxicity for Mosquitoes and Fruit Flies

The continued use of insecticides for public health and agriculture has led to widespread insecticide resistance and hampering of control methods. Insecticide resistance surveillance of mosquito populations is typically done through Centers for Disease Control and Prevention (CDC) bottle bioassays or World Health Organization (WHO) tube tests. However, these methods can result in a high degree of variability in mortality data due to variable insecticide contact with&#160;the insect, the relatively small numbers of organisms tested, extensive variation in mass between populations, and constantly changing environmental conditions, leading to variable outcomes. This paper presents the topical application bioassay, adapted as a high-throughput phenotypic bioassay for both mosquitoes and fruit flies, to test large numbers of insects along&#160;a range of insecticide concentrations.
This assay 1) ensures consistent treatment and insecticide contact with every organism, 2) produces highly specific dose-response curves that account for differences in average mass between strains and sexes (which is particularly important for field-collected organisms), and 3) allows for the calculation of statistically rigorous median lethal doses (LD50), which are necessary for resistance ratio comparisons-an alternative surveillance approach from diagnostic dose mortality, which is also used for larvicide resistance surveillance. This assay will be a complementary tool for accurately phenotyping mosquito populations and, as illustrated using fruit flies, is easily adaptable for use with other insects. We argue that this assay will help fill the gap between genotypic and phenotypic insecticide resistance in multiple insect species.

We describe the methodology and importance of the topical application bioassay to measure insecticide susceptibility in mosquitoes and fruit flies. The presented assay is high-throughput, utilizes insect mass-thus allowing for calculating a mass-relativized lethal dose instead of concentration-and likely has lower variability than other similar methods.

局部应用生物测定以量化蚊子和果蝇的杀虫剂毒性

The continued use of insecticides for public health and agriculture has led to widespread insecticide resistance and hampering of control methods. ...

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

Native to East Asia, the spotted-wing drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), has established widely in the Americas, Europe, and parts of Africa over the last decade, becoming a devastating pest of various soft-skinned fruits in its invaded regions. Biological control, especially by means of self-perpetuating and specialized parasitoids, is expected to be a viable option for sustainable area-wide management of this highly mobile and polyphagous pest. Ganaspis brasiliensis Ihering (Hymenoptera: Figitidae) is a larval parasitoid that is widely distributed in East Asia, and has been found to be one of the most effective parasitoids of D. suzukii.
Following rigorous pre-introduction evaluations of its efficacy and potential non-target risks, one of the more host-specific genetic groups of this species (G1 G. brasiliensis) has been approved recently for introduction and field release in the United States and Italy. Another genetic group (G3 G. brasiliensis), which was also commonly found to attack D. suzukii in East Asia, may be considered for introduction in the near future. There is currently enormous interest in rearing G. brasiliensis for research or in mass-production for field release against D. suzukii. This protocol and associated video article describe effective rearing methods for this parasitoid, both on a small scale for research and a large scale for mass-production and field release. These methods may benefit further long-term research and use of this Asian-native parasitoid as a promising biological control agent for this global invasive pest.

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

Ganaspis brasiliensis-a larval parasitoid of Drosophila suzukii (a global invasive fruit crop pest)-has been approved or is considered for introduction into Europe and the United States for biological control of this pest. This article provides protocols for both small-scale and large-scale rearing of this parasitoid.

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, ...

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

摘要

探索更多视频

此视频中的章节

学习在果蝇的侵略（果蝇）

蚊子养育的议定书“（冈比亚按蚊）

KLRG1四聚体的生产协议

果实角质层分离和生物物理研究

果实挥发分析，利用电子鼻

视网膜平片准备成像和免疫组化优化方案

稳定蝇及其他 Muscoid 双翅目的实验室饲养

简单的自制工具处理果蝇**果蝇黑色素*

局部应用生物测定以量化蚊子和果蝇的杀虫剂毒性

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

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

摘要

探索更多视频

此视频中的章节

学习在果蝇的侵略（果蝇）

蚊子养育的议定书“（冈比亚按蚊）

KLRG1四聚体的生产协议

果实角质层分离和生物物理研究

果实挥发分析，利用电子鼻

视网膜平片准备成像和免疫组化优化方案

稳定蝇及其他 Muscoid 双翅目的实验室饲养

简单的自制工具处理果蝇*果蝇黑色素

局部应用生物测定以量化蚊子和果蝇的杀虫剂毒性

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

简单的自制工具处理果蝇**果蝇黑色素*

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