我们要感谢巴西比奥费布里卡·莫斯卡梅德、帕斯基萨·多·埃斯塔多·德圣保罗基金会（FAPESP）和特克诺洛尼亚国家委员会（CNPq）的财政支持。 我们还要感谢以下人员的援助：米里亚姆·多斯桑托斯、吉尔迪安·席尔瓦、格西兰·多斯桑托斯、法比奥·戈纳尔维斯、约翰·保罗·奥利维拉、路易莎·加尔齐拉、何塞·卡洛斯·瓦伦尼亚。

昆虫要求
1. 概述
<ol><li>大规模饲养有几个阶段，每个阶段都在大规模饲养单元的不同区域进行。 图1 显示了饲养的主要阶段和它们发生在大规模饲养设施中的位置。</li>
</ol>2. 昆虫的生物安全考虑
<ol><li>该项目得到了巴西圣保罗大学动物伦理委员会的批准，并获得了所有必要的许可证和批准。</li>
	<li>RIDL 系统使用有条件的杀伤力：膳食补充剂（四环素）可防止RIDL系统的表达，使昆虫大量饲养：没有四环素蚊子就会死亡。在野外的 Ae.aegypti 繁殖地没有发现四环素，因此任何从逃亡者身上产生的后代都无法存活。 这比使用用其他方法（即 辐射）消毒的野生型（WT）昆虫增加了额外的安全性。</li>
	<li>使用双门入口，"仅限授权人员"标志和昆虫屏幕中的昆虫。任何窗户都应密封，以避免WT昆虫污染该菌落，并有助于防止昆虫从内部逃逸。</li>
	<li>地方法规和当局可能有具体的其他或替代性要求。尽早与有关当局联系以获取信息/指导，并在昆虫中建立 RIDL 菌株之前获得所有必要的许可和批准。</li>
	<li>大规模饲养昆虫只应用于RIDL饲养。不应存在其他菌株或昆虫。这是为了避免对RIDL菌株的污染和对可能影响健康和生存的病原体的污染，降低被释放的男性的生产力和有效性。</li>
</ol>3. 昆虫设计
<ol><li>
图2 显示了巴西比奥菲布里卡·莫斯卡梅德使用的昆虫计划。</li>
	<li>所有房间的温度为26°C±2），相对湿度约为70-80%，光照时间为12：12：使用荧光灯条的暗周期。</li>
	<li>此蓝图可根据特定空间要求进行修改， 例如 释放规模将决定总体大小。然而，鸡蛋的生产、饲养和分拣、质量控制（QC）和释放应分为四个不同的区域：鸡蛋生产殖民地、释放生成、QC和成人储存放生。</li>
	<li>鸡蛋生产殖民地室（约20米2）用于生产卵子的释放生成。</li>
	<li>质量控制区是进行实验，检查程序和RIDL线的适合性，包括测量舱口率，幼崽数量，幼崽大小，测量分拣和移除女性的有效性。还通过使用测量杀伤力的偏移检测来检查荧光标记和表型是否存在，从而对WT污染进行了测试。</li>
	<li>释放生成室（约36米2）需要足够的空间来整理托盘和幼虫和幼虫。托盘存放在专用铝架上。饲养和分拣需要一个大的工作区域和水槽，洗盘和笼子需要另一个大水槽。水槽的废水口中包括了一个过滤器捕获陷阱，以减轻活蚊子通过下水道的逃逸。</li>
	<li>托盘上使用的所有水都通过排水沟处理，排水沟有一个装有网格的隔间，可防止昆虫在任何发育阶段逃逸。</li>
	<li>成人的释放存储区域（成人存储释放， 图 2），包含机架以容纳释放设备，而成人在释放前成熟。</li>
</ol>大规模饲养 RIDL 的生产方法：
蛋生产殖民地生产用于释放代（图1）的鸡蛋，以生产成人。这两种饲养方法有许多相似之处，但也有一些明显的区别。仅在鸡蛋生产殖民地部分描述两个程序相同的饲养过程。
鸡蛋生产殖民地
4. 拉瓦尔生产
鸡蛋生产殖民地产生同源OX513A RIDL鸡蛋8 为释放一代。高质量的控制确保了所供应鸡蛋的生存能力、体能和应变完整性。
<ol><li>孵化的刺激是浸入低溶解氧的水中。为了降低水中的氧气含量，加热水，直到它沸腾，并立即将400毫升放入500毫升玻璃罐（74毫米开口周），牢牢地固定盖子，并在室温下离开几个小时冷却。</li>
	<li>将1克鸡蛋放入一罐白开水中，重新seal，等待一小时。将物品转移到2升水的托盘中，在昆虫条件下过夜。</li>
	<li>将孵化的幼虫放入已知的水量中，用磁搅拌器和跳蚤搅拌，以获得足够的时间取回松石：剧烈搅拌或长时间伤害幼虫，应保持在最低限度。用于孵化幼虫的标准体积为 1 L，但超过 300 幼虫/毫升的密度难以计数，而且可能具有破坏性。因此，孵化不超过30万个鸡蛋/升的水。</li>
	<li>通过取三个 1 毫升的吸水纸，并在吸水海绵上放置一张 1 厘米方格的吸水纸，以吸收多余的水，确定舱位率。 通过寻找孵化鸡蛋的缺失帽，从三个正方形计算孵化和无帽子的鸡蛋数量。预计孵化率约为80-90%。</li>
	<li>取四个1毫升的松饼，放入四个黑色称重船（幼虫是白色的，因此在黑色背景下更容易看到），并计算眼睛呈现的活幼虫的数量：建议使用计数器。然后，每毫升幼虫的平均数量用于计算每个托盘上添加的水量，以获得每个托盘所需的幼虫数量。</li>
	<li>幼虫在株和物种之间饲养的密度可能有很大的差异。将RIDL成年雄性与野生型雄性进行比较也是可取的：理想情况下，你想要类似于更大的RIDL雄性，可以成功地与野生雄性竞争。因此，对于每个 RIDL 菌株，需要确定幼虫的理想密度，并权衡饲养的空间限制和生产规模。密度从0.1-2.5幼虫/毫升，我们发现是生产产出和质量的良好组合：也讨论了美第奇23。</li>
	<li>将幼虫添加到托盘中。在我们的设施中，我们使用约 53 厘米 x 37 厘米 x 8 厘米（L x W x H） 的托盘和所需的水量，以达到幼虫的密度以进行生产。</li>
	<li>在 1：100 稀释时将四环素（水中 3 毫克/毫升）的库存溶液添加到托盘中，以获得 30μg/ml 所需的最终浓度。</li>
	<li>每天用片状鱼食<a href="http://www.sera.de">（www.sera.de）</a>喂幼虫，这些幼虫已被压成细粉。 表1 显示了典型的每天每幼虫食物毫克的喂养制度。在上述昆虫条件下喂养幼虫，直到适当的排序日。</li>
</ol>5. 从普佩和男性/女性普佩分拣拉瓦
<ol><li>通过将幼虫与幼虫分离，在孵化8天后从幼虫中分离出幼虫。然后对男性进行性排序。一种称为板分离器24，25 的装置可以从雄性幼虫和雌性幼虫中分拣幼虫。 图3 说明了使用这个板分离器将幼虫从雄性幼虫和雌性幼虫中分离出来。</li>
	<li>使用底部有网状物的测量勺，用 500 pupae 或更多校准它，并用它来设置笼子，并估计产生的雄性和雌性 pupae 的总数量。</li>
	<li>在分拣的第一天（第8天），大多数幼崽是雄性，所以把分类的幼虫放回托盘，然后饲养到第二天，当大多数幼崽是雌性（图4）。</li>
	<li>每天重复步骤 2.1-2.3 中描述的排序过程，直到所有幼虫都吐出或直到工作周结束。</li>
	<li>将 1，000 只雄性幼崽（使用测量勺子）放在笼子里，第 9 天在同一笼子里添加 3，000 只雌性幼崽（我们使用 30 厘米高、直径 30 厘米的 PVC 管，配有网状顶部和网状通道孔，但 BugDorm 也提供 30 厘米 x 30 厘米 x 30 厘米塑料笼）。</li>
	<li>允许成人交配至少2天，并为他们提供蔗糖溶液（10%）在血液喂养前在湿棉 广告利比图姆 。收集鸡蛋两周（~两个淋病循环），每周两次血液喂养。这平均产生143，000个卵子从3，000个女性的笼子里（平均48个卵子/雌）。每周生产400万只鸡蛋，每周需要设置28个笼子。</li>
</ol>6. 血液喂养
<ol><li>铝板喂养系统每周两次为笼子提供血液。铝板（10 厘米 x 10 厘米 x 3 毫米）侧面用 Parafilm （图 5）创建一个血袋。为了鼓励喂养，血液通过在板材喂食器上放置一个加热豆袋来加热。豆袋由布袋中约250克小麦粒组成，在微波炉中加热约10秒。我们用来喂养的血液来自当地的屠宰场：由于这些动物（主要是山羊和绵羊）是供人类食用的，因此它们经过测试，以避免病原体的存在。</li>
	<li>喂血三天后，为雌性产卵提供了一个输卵部位。渗透部位是一个圆形塑料容器，里面装满了约1/4的水和滤纸。两天后，渗漏位置被移除。为了最大限度地生产鸡蛋，请确保滤纸覆盖容器内的整个可用表面，并保持湿润。</li>
	<li>取出蛋纸，放在吸尿纸上，在昆虫条件下晾干：鸡蛋需要一段时间的调理在高湿度（超过70%）奠定后至少48小时。只要湿度保持在25度以上，鸡蛋就可以在昆虫中保留长达3个月，孵化率最小降低。</li>
	<li>鸡蛋生产群落需要足够大小，以提供每周为释放计划所需的鸡蛋数量。上述设施可生产约400万个鸡蛋，每周设置28个笼子。建议保留足够的鸡蛋储存，以保证至少4周的鸡蛋释放生成和鸡蛋生产殖民地。</li>
</ol>发布生成
7. 拉瓦尔生产
释放生成的孵化、饲养和分拣过程与先前描述的鸡蛋生产群落方法相同。然而，生产规模要大得多，这需要更多的托盘和更多的努力。
8. 分拣拉维、雄性普佩和雌性普佩
<ol><li>幼虫、雄性幼崽和雌性幼崽的分类与蛋生产描述的方法相同。</li>
	<li>对雄性幼崽进行分类后，在释放前检查雌性污染物是很重要的：在场的女性不应超过1%。取三个500 pupae的阿利奎特（三个随机的小狗测量勺子），并计算在场的雌性小狗的数量。女性可以识别，因为他们一般比男性大，从生殖器叶形状的差异（图6）27。如果女性超过1%，则应再次求助并检查她们。</li>
	<li>仅使用第8天和第9天的男性释放：高压灭菌任何剩余的幼虫和幼虫。在巴西，这种废物必须与医疗废物一样对待，但其他国家的法规可能有所不同。不要使用卵子生产殖民地释放代的雌性幼崽和幼虫，因为质量控制、饲养和污染风险的差异来自数量较大，质量控制较低。</li>
</ol>成人存储释放：将男性放入释放设备中，在发布前出现并成熟。此方法不包括释放设备的详细信息。但是，我们使用的释放设备的容量约为 1，000 男性。此方法不包括释放方法详细信息（释放设备和释放系统）。

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Entomol. 97, 1547-1553 (2004).</li><li>Papathanos, P. 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=Sex+separation+strategies:+past+experience+and+new+approaches.">Sex separation strategies: past experience and new approaches.</a> Malar. J.. 8 Suppl 2 (S5), (2009).</li><li>Medici, 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=Studies+on+Aedes+albopictus+larval+mass-rearing+optimization.">Studies on Aedes albopictus larval mass-rearing optimization.</a> J. Econ. Entomol. 104, 266-273 (2011).</li><li>Focks, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+improved+separator+for+the+developmental+stages,+sexes+and+species+of+mosquito+(Diptera+Culicidae)..">An improved separator for the developmental stages, sexes and species of mosquito (Diptera Culicidae)..</a> J. Med. Entomol. 17, 567-568 (1980).</li><li>Fay, R. W., McCray, E. M., Kilpatrick, 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=Mass+production+of+sterilized+male+Aedes+aegypti.">Mass production of sterilized male Aedes aegypti.</a> Mosquito News. 23, 210-214 (1963).</li><li>Christophers, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Aedes aegypti the yellow fever mosquito: Its life history, Bionomics and Structure. , (2009).</li><li>Jones, 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=A+simple+method+for+sexing+living+Anopheles+Larvae+(diptera,+culicidae).">A simple method for sexing living Anopheles Larvae (diptera, culicidae).</a> Ann. Entomol. Soc. Am. 50, 104-106 (1957).</li><li>Koenraadt, C. J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pupal+Dimensions+as+Predictors+of+Adult+Size+in+Fitness+Studies+of+Aedes+aegypti+(Diptera+Culicidae)..">Pupal Dimensions as Predictors of Adult Size in Fitness Studies of Aedes aegypti (Diptera Culicidae)..</a> J. Med. Entomol. 45, 331-336 (2008).</li><li>Alphey, L., Nimmo, D., O'Connell, S., Alphey, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insect+population+suppression+using+engineered+insects.">Insect population suppression using engineered insects.</a> Adv. Exp. Med. Biol. 627, 93-103 (2008).</li><li>Alphey, N., Bonsall, M. B., Alphey, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modeling+resistance+to+genetic+control+of+insects.">Modeling resistance to genetic control of insects.</a> J. Theor. Biol. 270, 42-55 (2011).</li><li>Atkinson, M. 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=Analyzing+the+control+of+mosquito-borne+diseases+by+a+dominant+lethal+genetic+system.">Analyzing the control of mosquito-borne diseases by a dominant lethal genetic system.</a> Proc. Natl. Acad. Sci. U.S.A. 104, 9540-9545 (2007).</li><li>Bargielowski, I., Nimmo, D., Alphey, L., Koella, 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=Comparison+of+life+history+characteristics+of+the+genetically+modified+OX513A+line+and+a+wild+type+strain+of+Aedes+aegypti.">Comparison of life history characteristics of the genetically modified OX513A line and a wild type strain of Aedes aegypti.</a> PLoS One. 6 (e20699), (2011).</li><li>Bargielowski, I., Alphey, L., Koella, 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=Cost+of+mating+and+insemination+capacity+of+a+genetically+modified+mosquito+Aedes+aegypti+OX513A+compared+to+its+wild+type+counterpart.">Cost of mating and insemination capacity of a genetically modified mosquito Aedes aegypti OX513A compared to its wild type counterpart.</a> PLoS One. 6 (26086), (2011).</li></ol>

奥克斯泰克的作者拥有Oxitec有限公司和牛津大学的就业和/或股权，持有与本文主题相关的知识产权。

RIDL是一种有效和环保的方法，控制蚊子3，29-31。该技术适用于综合害虫管理计划，目前大多数控制方法，包括杀幼虫剂、繁殖地减少和成人杀虫剂都与这项技术相容。此方法描述了如何每周生产多达 570，000 个 RIDL 雄性幼崽，用于 Ae 的控制。据我们所知，这是第一次描述这种规模的转基因蚊子的生产。在20世纪60年代和70年代25年代，为野生Ae.aegypti开发了一些类似的生产系统，但自那时以来，一直没有这种规模的可比生产。从2011年2月到2012年2月，巴西约有1100万男性被释放。控制某一区域所需的雄性数量取决于若干因素，包括野生种群的规模、释放的雄性分布、释放后雄性生存和交配竞争力以及环境条件。 先前的研究表明，RIDL至少可以减少80%的蚊子数量。
在优化生产规模和成本与男性质量之间需要平衡。例如，增加幼虫密度可以通过减少所需的空间、劳动力和时间来增加生产能力。然而，幼虫密度过高会导致寿命更小、寿命较短的雄配能力降低32 ， 33 。与 SIT 计划相关的男性质量最终将由被释放的男性在实地与女配的能力来评估。需要广泛的实地评估，以评估交配竞争力相对于野生同行9，10。这往往使准确评估哪些因素使"高质量"雄性蚊子不切实际。然而，在大规模养殖中保持一致的生产和质量（在可以定期评估的程度）是至关重要的。这需要高度警惕和标准化所有流程，小波动可能会产生重大影响。L1幼虫的引文是关键一步，说明了这一点。将正确数量的幼虫加入托盘对优质生产至关重要。喂养制度正是为特定数量的幼虫量身定做的。幼虫太少/太多会导致过度/喂养不足，从而影响幼虫的生存、幼虫的大小和幼崽的时间。如果说大规模饲养艺术有一个秘密的话，就是要确保生产周期中的许多小步骤能够像本文所述，持续、精确、高质量地进行。

蚊子传播许多病原体，这些病原体可引起人类一系列疾病，控制这些蚊子几个世纪以来一直是一场持续的战斗。基于化学和生物方法控制昆虫的战略取得了一些显著成功，但在许多情况下，控制在长期内是不可持续的。例如，巴西在50年代实现了消灭 Ae.aegypti， 但蚊子在过去40-50年中重新入侵。这可以归因于许多原因，包括杀虫剂耐药性，环境破坏，控制方案实施不力15-20，以及快速再入侵没有充分的监测或反应。
正在寻求新的技术和方法，试图赢得这场与蚊子的战斗。 分子技术的最新进展导致以无菌昆虫技术（SIT）1-3为基础的新的和创新的蚊虫控制方法的发展。一种称为RIDL（携带主要致命物的昆虫释放）4的控制方法以SIT为基础，但使用遗传方法消除辐射灭菌的需要。已为包括Ae.aegypti 5-8、Ae.aegypti的RIDL菌株在内的多种害虫品种建造了RIDL菌株，在大开曼9，10油田进行了成功测试：计划或正在世界其他国家进一步实地使用。 使用RIDL抑制蚊子种群需要饲养大量高质量的男性成年人。
对于 SIT 程序，只释放男性被认为是可取的;雌性蚊子叮咬和传播疾病。此外，被释放的无菌男性可能会被被释放的女性"分心"，降低项目的有效性。在辐照地中海果蝇21的大型野外实验中，雄性释放比混合性别释放有效3-5倍。
在 RIDL 大规模饲养计划中，有几个阶段可以生产雄性进行释放。 第一种是生产释放生成所需的鸡蛋（图1）。下一阶段是饲养卵子到幼崽或成人，将幼虫与幼虫和雄性幼崽从雌性幼崽中分离出来。在适合大规模分类的特定生命阶段，男性与女性的大规模分离需要性别差异。在Ae.aegypti（和其他蚊子物种）中，雄性与雌性幼虫之间存在显著的大小差异，可以利用这种差异进行性别分离技术。排序的RIDL男性然后被释放在控制程序作为脓包或成人11，12。
以下描述了OX513A的大规模饲养方法，OX513A是一种E的RIDL菌株 。艾吉普提， 要释放所述方法涵盖了生产卵子和用于控制程序的RIDL雄性所需的技术。

<table align="left" border="1" cellpadding="0" cellspacing="0" width="100%">
	<tbody>
		<tr>
			<td style="width:15.92%;height:38px;">
			Vipan Premium
			</td>
			<td style="width:15.06%;height:38px;">
			Sera GmbH
			</td>
			<td style="width:12.64%;height:38px;">
			190
			</td>
			<td style="width:40.2%;height:38px;">
			<a href="http://www.sera.de/uk/pages/products/product/sera-vipan-3.html">http://www.sera.de/uk/pages/products/product/sera-vipan-3.html</a>
			</td>
			<td style="width:16.16%;height:38px;">
			Required for rearing RIDL larvae
			</td>
		</tr>
		<tr>
			<td style="width:15.92%;height:58px;">
			Tetracycline
			</td>
			<td style="width:15.06%;height:58px;">
			Sigma Aldrich
			</td>
			<td style="width:12.64%;height:58px;">
			T7660
			</td>
			<td style="width:40.2%;height:58px;">
			<a href="http://www.sigmaaldrich.com/catalog/ProductDetail.do?D7=0&amp;N5=SEARCH_CONCAT_PNO|BRAND_KEY&amp;N4=T7660|SIGMA&amp;N25=0&amp;QS=ON&amp;F=SPEC">http://www.sigmaaldrich.com/catalog/ProductDetail.do?D7=0&#38;N5=SEARCH_CONCAT_PNO|BRAND_KEY&#38;N4=T7660|SIGMA&#38;N25=0&#38;QS=ON&#38;F=SPEC</a>
			</td>
			<td style="width:16.16%;height:58px;">
			Required for rearing RIDL larvae
			</td>
		</tr>
		<tr>
			<td style="width:15.92%;height:38px;">
			Plate separator
			</td>
			<td style="width:15.06%;height:38px;">
			J.W. Hock
			</td>
			<td style="width:12.64%;height:38px;">
			5412
			</td>
			<td style="width:40.2%;height:38px;">
			<a href="http://www.johnwhock.com/download/manuals/instr_5412_separator.pdf">http://www.johnwhock.com/download/manuals/instr_5412_separator.pdf</a>
			</td>
			<td style="width:16.16%;height:38px;">
			Separating larvae and pupae
			</td>
		</tr>
		<tr>
			<td style="width:15.92%;height:38px;">
			Parafilm M
			</td>
			<td style="width:15.06%;height:38px;">
			Pechiney Plastic packing
			</td>
			<td style="width:12.64%;height:38px;">
			PM-996
			</td>
			<td style="width:40.2%;height:38px;">
			<a href="http://www.parafilm.com">www.parafilm.com</a>
			</td>
			<td style="width:16.16%;height:38px;">
			Cover plate for blood feeding system
			</td>
		</tr>
		<tr>
			<td style="width:15.92%;height:38px;">
			Rearing pans for Release generation (53 cm x 38 cm x 8 cm)
			</td>
			<td style="width:15.06%;height:38px;">
			Pleion
			</td>
			<td style="width:12.64%;height:38px;">
			0757
			</td>
			<td nowrap="nowrap" style="width:40.2%;height:38px;">
			<a href="http://pleion.actcenter.com.br/produtos.asp?opcao=1">http://pleion.actcenter.com.br/produtos.asp?opcao=1</a>
			</td>
			<td style="width:16.16%;height:38px;">
			Larval rearing Release generation
			</td>
		</tr>
		<tr>
			<td style="width:15.92%;height:19px;">
			Fluorescent scope
			</td>
			<td style="width:15.06%;height:19px;">
			Leica&#160;Microsystems
			</td>
			<td style="width:12.64%;height:19px;">
			MZ FLIII
			
			</td>
			<td style="width:40.2%;height:19px;">
			<a href="http://www.leica-microsystems.com/fileadmin/downloads/Leica%20MZ%20FLIII/Brochures/M1-160-0de.pdf">http://www.leica-microsystems.com/fileadmin/downloads/Leica%20MZ%20FLIII/Brochures/M1-160-0de.pdf</a>
			</td>
			<td style="width:16.16%;height:19px;">
			Viewing fluorescent RIDL larvae
			</td>
		</tr>
		<tr>
			<td style="width:15.92%;height:38px;">
			Adult cages
			</td>
			<td style="width:15.06%;height:38px;">
			BugDorm
			</td>
			<td style="width:12.64%;height:38px;">
			DP1000
			</td>
			<td style="width:40.2%;height:38px;">
			<a href="http://bugdorm.megaview.com.tw/bugdorm-1-insect-rearing-cage-30x30x30-cm-pack-of-one-p-29.html">http://bugdorm.megaview.com.tw/bugdorm-1-insect-rearing-cage-30x30x30-cm-pack-of-one-p-29.html</a>
			</td>
			<td style="width:16.16%;height:38px;">
			Cages for Egg production&#160; colony
			</td>
		</tr>
		<tr>
			<td style="width:15.92%;height:38px;">
			Filter paper
			</td>
			<td style="width:15.06%;height:38px;">
			CELAB
			</td>
			<td style="width:12.64%;height:38px;">
			
			</td>
			<td style="width:40.2%;height:38px;">
			<a href="http://www.casadolaboratorio.com.br/subpage118.html">http://www.casadolaboratorio.com.br/subpage118.html</a>
			</td>
			<td style="width:16.16%;height:38px;">
			Filter paper for egg laying
			</td>
		</tr>
	</tbody>
</table>

mass production of genetically modified aedes aegypti for field releases in brazil

正在寻求新的技术和方法，以争取赢得与蚊子的战斗。分子技术的最新进展导致以无菌昆虫技术（SIT）1-3为基础的新的和创新的蚊虫控制方法的发展。一种称为RIDL（携带主要致命物的昆虫释放）4的控制方法，以SIT为基础，但使用遗传方法消除辐射灭菌5-8的需要。在大开曼9，10的田地里，一种E.aegypti的RIDL菌株被成功测试：计划或正在世界其他国家进一步实地使用。
在几个昆虫物种中已经建立了大规模昆虫饲养，每周达到数十亿的水平。然而，在蚊子中，饲养一般规模要小得多，大多数大规模饲养是在1970年代和80年代进行的。对于 RIDL 计划，最好尽可能少地释放女性，因为它们会咬伤和传播疾病。在大规模饲养计划中，要释放雄性有几个阶段：产卵、饲养卵子直到幼崽，然后在释放前从雌性中分拣雄性。然后，这些雄性用于RIDL控制程序，释放为幼崽或成人11，12。
要抑制蚊子种群使用RIDL大量的高素质男性成人需要饲养13，14。以下描述了OX513A的大规模饲养方法，OX513A是Ae.aegypti 8的一种REL菌株，用于释放，并涵盖了生产卵子和大规模饲养REL雄性进行控制程序所需的技术。

预期的幼崽生产结果显示在 图4中。雄性幼崽先，第8天达到顶峰，孵化后第9天雌性达到高峰。重要的是要测量这个小狗曲线，知道最好的时间排序男性从女性。
超过6个月的男性和女性幼崽分类结果显示，女性平均污染率为0.02%（图5：SEM = 0.004%） 。这种污染率仅代表在一个月内释放的400名女性（约200万男性）。目前每周的鸡蛋产量为400万。其中350万只孵化用于卵子群落和生产，其余储存起来供备份。大约11%的鸡蛋用于卵子生产群落，其余用于释放生成。鸡蛋孵化率平均为87.3%（SEM = 0.5%）L1幼虫的雄性幼崽平均产量为29.5%（SEM =1.2）。每周为繁殖一代繁殖的幼虫饲养200万只，产自约571，000只雄性幼崽（SEM = 14，000只）。Pupae 和成人在出现和释放期间的平均死亡率为 5%，导致每周约有 543，000 只（SEM = 13，000）释放成年雄性蚊子。预计目前400万鸡蛋/周的产量可降至300万/周，并进一步优化，以减少浪费和储存造成的损失。所述生产总共需要6名工作人员：四个工作释放生成和其余的两个在卵子殖民地。

质量管理

质量控制对于保持成年人的素质、饲养效率、劳动力和成本至关重要。 

转基因表型控制
为了验证RIDL基因表达，我们进行了两项控制，一是检查荧光标记的表达，二是在没有四环素的情况下检查RIDL杀伤性特征的表达。所有幼虫都应该表达荧光标记。为了检查荧光标记是否按预期表示，2，000 只首次星内幼虫在 Leica MZFLIII 立体显微镜下使用 DsRed2 过滤器 （得克萨斯红） 进行检查，以确定是否有任何无荧光个体存在。没有四环素的RIDL基因被表达出来，我们预计成人8的存活率为3-4%。为了检查这一点，为每个未四环素的释放批次设置一个额外的托盘。与正常饲养的唯一区别是，食物从第6天起减少2/3，因为由于死幼虫积累多余的食物会导致细菌过度生长。

饲养控制
随机选择释放生成中的四个托盘进行排序和单独计数。每天从每个托盘记录的男性和女性幼崽的数量（图4）。与预期的男性和女性数量的任何偏差都表明存在一个潜在的问题，可能会影响生产和/或健身，并可与卵子群进行比较，以帮助确定问题的根源。

普佩测量
普佩的大小已被证明与成人大小28相关。作为成人尺寸的质量控制，从释放生成开始，每分拣一天至少测量 30 只雄性幼崽：对于鸡蛋生产殖民地女性幼崽也测量。释放的雄性的平均头孢菌宽度为1.05毫米（SEM 0.005），释放的雄性平均宽度为1.04毫米（SEM =0.006）和1.29毫米（SEM =0.006），男性和女性分别为卵子生产、殖民地：类似的结果也取得了类似的结果，为A.阿尔博皮图斯大规模饲养24。
<img alt="Figure 1" fo:content-width="5in" fo:src="/files/ftp_upload/3579/3579fig1highres.jpg" src="/files/ftp_upload/3579/3579fig1.jpg"> 图1。大规模饲养蚊子用于RIDL/SIT计划的阶段。 蛋类生产殖民地具有高水平的质量控制，以确保提供给释放一代的鸡蛋的质量。 卵子被饲养到释放的殖民地，在那里雄性从雌性排序。 然后，男性成人用于 RIDL 控制程序。
<img alt="Figure 2" fo:content-width="5in" fo:src="/files/ftp_upload/3579/3579fig2highres.jpg" src="/files/ftp_upload/3579/3579fig2.jpg"> 图2。 用于生产 RIDL 雄性为释放程序的饲养设施的示意图。 优质鸡蛋在鸡蛋生产殖民地室不断生产，然后通过饲养到释放生成室的幼蛋。幼虫和幼崽然后分开，幼崽为雄性进行性分类。然后，男性被放入释放装置，并允许成熟成人在成人存储和释放室释放。
<img alt="Figure 3" fo:content-width="5in" fo:src="/files/ftp_upload/3579/3579fig3highres.jpg" src="/files/ftp_upload/3579/3579fig3.jpg"> 图3。 使用板分离器26分离幼虫、雄性幼崽和雌性幼崽。  板块分离器使用幼虫、雄性幼崽和雌性幼崽之间的大小差异来分拣这三个不同的生命阶段：幼虫往往比雄性幼崽小，而雄性幼崽又比雌性幼崽小。 仪器由两个玻璃板组成：一个固定在第一个板的顶部，另一个可以移动相对于第一个使用四个调整螺丝（A）。 四个调整螺钉允许将外板设置在后板的角度，以便在板之间形成楔形空间，向下逐渐减少。 幼虫和幼虫倒在玻璃板（B）之间，用水管（C）轻轻清洗。 通过调整板的角度，幼虫、雄性幼崽和雌性幼崽可以分开（D）。 随着连续冲洗和增加板角，幼虫可以冲过第一个（进入筛子），其次是雄性幼崽，最后是雌性幼崽。
<img alt="Figure 4" fo:content-width="5in" fo:src="/files/ftp_upload/3579/3579fig4highres.jpg" src="/files/ftp_upload/3579/3579fig4.jpg"> 图4。 大规模饲养的里德尔Ae.艾吉普蒂的小狗曲线。  此图显示了在 23 周观察期间大规模饲养的 RIDL 雄性和雌性幼崽的平均比例，每周恢复约 135，000 只幼崽。 错误条=平均标准误差，n=23。在第一次收集（第8天），我们平均恢复59%的男性和30%的女性幼崽从总脓包从托盘恢复超过5天。
<img alt="Figure 5" fo:content-width="5in" fo:src="/files/ftp_upload/3579/3579fig5highres.jpg" src="/files/ftp_upload/3579/3579fig5.jpg"> 图5。 平均女性污染排序的雄性幼崽。 此图显示了在孵化超过六个月后，男性在第 8 天分拣期间女性污染的月平均百分比。
<img alt="Figure 6" fo:content-width="5in" fo:src="/files/ftp_upload/3579/3579fig6highres.jpg" src="/files/ftp_upload/3579/3579fig6.jpg"> 图6。铝板血液馈线系统。盘子（B） 覆盖着帕拉菲尔姆（A），血液被吹进口袋里， 然后密封（D）。盘子被放在笼子上，通过在上面放置一个加热的豆袋（C）加热。 
<img alt="Figure 7" fo:content-width="5in" fo:src="/files/ftp_upload/3579/3579fig7highres.jpg" src="/files/ftp_upload/3579/3579fig7.jpg"> 图7。 区分男性和女性 的艾吉普蒂 · 普佩。  Ae . aegypti 幼虫可以通过生殖器叶形状的差异（在桨下方的 pupal 腹部段的末端）可靠地进行。此外，男性也往往比女性小。
<img alt="Table 1" fo:content-width="5in" fo:src="/files/ftp_upload/3579/3579table1highres.jpg" src="/files/ftp_upload/3579/3579table1.jpg"> 表1。 Ae. 艾吉普蒂 里德尔幼虫的一般喂养制度（每天每幼虫的食物毫克）。 计算实际需要的食物量乘以每盘幼虫的总数。

Watch this Scientific Journal Video about Mass Production of Genetically Modified Aedes aegypti for Field Releases in Brazil at JoVE.com

Mass Production of Genetically Modified Aedes aegypti for Field Releases in Brazil

为了利用RIDL®（释放携带主要致命物的昆虫）系统来抑制伊蚊，需要释放大量的雄性蚊子。这就需要利用大规模饲养技术和工艺，提供可靠的系统，以获得数量最多的优质雄性蚊子。

巴西田间放生转基因 伊 蚊的大规模生产

New techniques and methods are being sought to try to win the battle against mosquitoes. Recent advances in molecular techniques have led to the ...

mass-production-genetically-modified-aedes-aegypti-for-field-releases

Research

JoVE Journal

Environment

Mass Production of Genetically Modified Aedes aegypti for Field Releases in Brazil

22.8K 次观看.  Oxitec Ltd. 为了利用RIDL®（释放携带主要致命物的昆虫）系统来抑制伊蚊，需要释放大量的雄性蚊子。这就需要利用大规模饲养技术和工艺，提供可靠的系统，以获得数量最多的优质雄性蚊子。

视频: Mass Production of Genetically Modified Aedes aegypti for Field Releases in Brazil

Evaluation of Integrated Anaerobic Digestion and Hydrothermal Carbonization for Bioenergy Production

Lignocellulosic biomass is one of the most abundant yet underutilized renewable energy resources. Both anaerobic digestion (AD) and hydrothermal carbonization (HTC) are promising technologies for bioenergy production from biomass in terms of biogas and HTC biochar, respectively. In this study, the combination of AD and HTC is proposed to increase overall bioenergy production. Wheat straw was anaerobically digested in a novel upflow anaerobic solid state reactor (UASS) in both mesophilic (37 &#176;C) and thermophilic (55 &#176;C) conditions. Wet digested from thermophilic AD was hydrothermally carbonized at 230 &#176;C for 6 hr for HTC biochar production. At thermophilic temperature, the UASS system yields an average of 165 LCH4/kgVS&#160;(VS: volatile solids) and 121 L CH4/kgVS&#160;at mesophilic AD over the continuous operation of 200 days. Meanwhile, 43.4 g of HTC biochar with 29.6 MJ/kgdry_biochar was obtained from HTC of 1 kg digestate (dry basis) from mesophilic AD. The combination of AD and HTC, in this particular set of experiment yield 13.2 MJ of energy per 1 kg of dry wheat straw, which is at least 20% higher than HTC alone and 60.2% higher than AD only.

A novel Upflow Anaerobic Solid State (UASS) reactor was used for biogas production from fibrous feedstock. Digestate from UASS reactor was hydrothermally carbonized into HTC biochar in a pressurized batch reactor. The integration of the two bioenergy concepts was applied in this study to increase overall bioenergy production.

评定一体化厌氧消化和水热碳化对生物能源生产

Lignocellulosic biomass is one of the most abundant yet underutilized renewable energy resources. Both anaerobic digestion (AD) and hydrothermal ...

科研

环境科学

Integrated Field Lysimetry and Porewater Sampling for Evaluation of Chemical Mobility in Soils and Established Vegetation

Potentially toxic chemicals are routinely applied to land to meet growing demands on waste management and food production, but the fate of these chemicals is often not well understood. Here we demonstrate an integrated field lysimetry and porewater sampling method for evaluating the mobility of chemicals applied to soils and established vegetation. Lysimeters, open columns made of metal or plastic, are driven into bareground or vegetated soils. Porewater samplers, which are commercially available and use vacuum to collect percolating soil water, are installed at predetermined depths within the lysimeters. At prearranged times following chemical application to experimental plots, porewater is collected, and lysimeters, containing soil and vegetation, are exhumed. By analyzing chemical concentrations in the lysimeter soil, vegetation, and porewater, downward leaching rates, soil retention capacities, and plant uptake for the chemical of interest may be quantified. Because field lysimetry and porewater sampling are conducted under natural environmental conditions and with minimal soil disturbance, derived results project real-case scenarios and provide valuable information for chemical management. As chemicals are increasingly applied to land worldwide, the described techniques may be utilized to determine whether applied chemicals pose adverse effects to human health or the environment.

Field lysimetry and porewater sampling allow researchers to evaluate the fate of chemicals applied to soils and established vegetation. The goal of this protocol is to demonstrate how to install required instrumentation and collect samples for chemical analysis during integrated field lysimetry and porewater sampling experiments.

综合外地Lysimetry和孔隙水采样的化学活动性土壤的评价和植被成立

Potentially toxic chemicals are routinely applied to land to meet growing demands on waste management and food production, but the fate of these chemicals ...

Extraction and Analysis of Microbial Phospholipid Fatty Acids in Soils

Phospholipid fatty acids (PLFAs) are key components of microbial cell membranes. The analysis of PLFAs extracted from soils can provide information about the overall structure of terrestrial microbial communities. PLFA profiling has been extensively used in a range of ecosystems as a biological index of overall soil quality, and as a quantitative indicator of soil response to land management and other environmental stressors.
The standard method presented here outlines four key steps: 1. lipid extraction from soil samples with a single-phase chloroform mixture, 2. fractionation using solid phase extraction columns to isolate phospholipids from other extracted lipids, 3. methanolysis of phospholipids to produce fatty acid methyl esters (FAMEs), and 4. FAME analysis by capillary gas chromatography using a flame ionization detector (GC-FID). Two standards are used, including 1,2-dinonadecanoyl-sn-glycero-3-phosphocholine (PC(19:0/19:0)) to assess the overall recovery of the extraction method, and methyl decanoate (MeC10:0) as an internal standard (ISTD) for the GC analysis.

Phospholipid fatty acids provide information about the structure of soil microbial communities. We present methods for extraction from soil samples with a single-phase chloroform mixture, fractionation of extracted lipids using solid phase extraction columns, and methanolysis to produce fatty acid methyl esters, which are analyzed by capillary gas chromatography.

提取及土壤微生物磷脂脂肪酸分析

Phospholipid fatty acids (PLFAs) are key components of microbial cell membranes. The analysis of PLFAs extracted from soils can provide information about ...

A Gusseted Thermogradient Table to Control Soil Temperatures for Evaluating Plant Growth and Monitoring Soil Processes

Thermogradient tables were first developed in the 1950s primarily to test seed germination over a range of temperatures simultaneously without using a series of incubators. A temperature gradient is passively established across the surface of the table between the heated and cooled ends and is lost quickly at distances above the surface. Since temperature is only controlled on the table surface, experiments are restricted to shallow containers, such as Petri dishes, placed on the table. Welding continuous aluminum vertical strips or gussets perpendicular to the surface of a table enables temperature control in depth via convective heat flow. Soil in the channels between gussets was maintained across a gradient of temperatures allowing a greater diversity of experimentation. The gusseted design was evaluated by germinating oat, lettuce, tomato, and melon seeds. Soil temperatures were monitored using individual, battery-powered dataloggers positioned across the table. LED lights installed in the lids or along the sides of the gradient table create a controlled temperature chamber where seedlings can be grown over a range of temperatures. The gusseted design enabled accurate determination of optimum temperatures for fastest germination rate and the highest percentage germination for each species. Germination information from gradient table experiments can help predict seed germination and seedling growth under the adverse soil conditions often encountered during field crop production. Temperature effects on seed germination, seedling growth, and soil ecology can be tested under controlled conditions in a laboratory using a gusseted thermogradient table.

Traditional thermogradient tables create a range of temperatures across the surface. Welding gussets perpendicular to the surface of a thermogradient table will control temperature in depth increasing possible research applications.

一个插角Thermogradient表来控制土壤温度为评估植物生长和监测土壤过程

Thermogradient tables were first developed in the 1950s primarily to test seed germination over a range of temperatures simultaneously without using a ...

Modification and Application of a Leaf Blower-vac for Field Sampling of Arthropods

Rice fields host a large diversity of arthropods, but investigating their population dynamics and interactions is challenging. Here we describe the modification and application of a leaf blower-vac for suction sampling of arthropod populations in rice. When used in combination with an enclosure, application of this sampling device provides absolute estimates of the populations of arthropods as numbers per standardized sampling area. The sampling efficiency depends critically on the sampling duration. In a mature rice crop, a two-minute sampling in an enclosure of 0.13 m2 yields more than 90% of the arthropod population. The device also allows sampling of arthropods dwelling on the water surface or the soil in rice paddies, but it is not suitable for sampling fast flying insects, such as predatory Odonata or larger hymenopterous parasitoids. The modified blower-vac is simple to construct, and cheaper and easier to handle than traditional suction sampling devices, such as D-vac. The low cost makes the modified blower-vac also accessible to researchers in developing countries.

Assessment of arthropod abundance in crops is critical for investigating population dynamics and species interactions. Here we describe the modification and application of a leaf blower-vac for suction sampling of arthropods in rice.

修改和叶鼓风机-VAC的节肢动物领域采样中的应用

Rice fields host a large diversity of arthropods, but investigating their population dynamics and interactions is challenging. Here we describe the ...

The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea

Plants possess a powerful system to defend themselves against potential threats by pathogenic fungi. For agriculturally important plants, however, current measures to combat such pathogens have proved too conservative and, thus, not sufficiently effective, and they can potentially pose environmental risks. Therefore, it is extremely necessary to identify host-resistance factors to assist in controlling plant diseases naturally through the identification of resistant germplasm, the isolation and characterization of resistance genes, and the molecular breeding of resistant cultivars. In this regard, there is need to establish an accurate, rapid, and large-scale inoculation method to breed and develop plant resistance genes. The rice blast fungal pathogen Magnaporthe grisea causes severe disease symptoms and yield losses. Recently, M. grisea has emerged as a model organism for studying the mechanisms of plant-fungal pathogen interactions. Hence, we report the development of a plant virulence test method that is specific for M. grisea. This method provides for both spray inoculation with a conidial suspension and wounding inoculation with mycelium cubes or droplets of conidial suspension. The key step of the wounding inoculation method for detached rice leaves is to make wounds on plant leaves, which avoids any interference caused by host penetration resistance. This spray/wounding protocol contributes to the rapid, accurate, and large-scale screening of the pathotypes of M. grisea isolates. This integrated and systematic plant infection method will serve as an excellent starting point for gaining a broad perspective of issues in plant pathology.

The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea

Here, we present a protocol to test plant virulence with the plant pathogen Magnaporthe grisea. This report will contribute to the large-scale screening of the pathotypes of fungi isolates and serve as an excellent starting point for understanding the resistant mechanisms of plants during molecular breeding.

植物感染试验: 以植物病原菌为载体的喷雾和伤口介导接种

Plants possess a powerful system to defend themselves against potential threats by pathogenic fungi. For agriculturally important plants, however, current ...

Laboratory and Field Protocol for Estimating Sheet Erosion Rates from Dendrogeomorphology

Sheet erosion is among the crucial drivers of soil degradation. Erosion is controlled by environmental factors and human activities, which often lead to severe environmental impacts. The understanding of sheet erosion is, consequently, a worldwide issue with implications for both environment and economies. However, the knowledge on how erosion evolves in space and time is still limited, as well as its effects on the environment. Below, we explain a new dendrogeomorphological protocol for deriving eroded soil thickness (Ex) by acquiring accurate microtopographic data using both terrestrial laser scanning (TLS) and microtopographic profile gauges. Additionally, standard dendrogeomorphic procedures, dependent on anatomical variations in root rings, are utilized to establish the timing of exposure. Both TLS and microtopographic profile gauges are used to obtain ground surface pro&#64257;les, from which Ex is estimated after the threshold distance (TD) is determined, i.e., the distance between the root and the sediment knickpoint, which allows de&#64257;ning the lowering of the ground surface caused by sheet erosion. For each profile, we measured the height between the topside of the root and a virtual plane tangential to the ground surface. In this way, we intended to avoid small-scale impacts of soil deformation, which may be due to pressures exerted by the root system, or by the arrangement of exposed roots. This may provoke small amounts of soil sedimentation or erosion depending on how they physically affect the surface runoff. We demonstrate that an adequate microtopographic characterization of exposed roots and their associated ground surface is very valuable to obtain accurate erosion rates. This finding could be utilized to develop the best management practices designed to eventually halt or perhaps, at least, lessen soil erosion, so that more sustainable management policies can be put into practice.

Characterizing erosion from dendrogeomorphology has usually focused on accurately finding the starting time of root exposure, by examining macroscopic or cell level changes caused by exposure. Here, we offer a detailed description of different novel techniques to obtain more precise erosion rates from highly accurate microtopographic data.

利用树形地貌估算板材侵蚀速率的实验室和现场协议

Sheet erosion is among the crucial drivers of soil degradation. Erosion is controlled by environmental factors and human activities, which often lead to ...

Construction of a Compact Low-Cost Radiation Shield for Air-Temperature Sensors in Ecological Field Studies

Low cost temperature sensors are increasingly used by ecologists to assess climatic variation and change on ecologically relevant scales. Although cost-effective, if not deployed with proper solar radiation shielding, the observations recorded from these sensors will be biased and inaccurate. Manufactured radiation shields are effective at minimizing this bias, but are expensive compared to the cost of these sensors. Here, we provide a detailed methodology for constructing a compact version of a previously described custom fabricated radiation shield, which is more accurate than other published shielding methods that attempt to minimize shield size or construction costs. The method requires very little material: corrugated plastic sheets, aluminum foil duct tape, and cable ties. One 15 cm and two 10 cm squares of corrugated plastic are used for each shield. After cutting, scoring, taping and stapling of the sheets, the 10 cm squares form the bottom two layers of the solar radiation shield, while the 15 cm square forms the top layer. The three sheets are held together with cable ties. This compact solar radiation shield can be suspended, or placed against any flat surface. Care must be taken to ensure that the shield is completely parallel to the ground to prevent direct solar radiation from reaching the sensor, possibly causing increased warm biases in sun-exposed sites in the morning and afternoon relative to the original, larger design. Even so, differences in recorded temperatures between the smaller, compact shield design and the original design were small (mean daytime bias = 0.06 &#176;C). Construction costs are less than half of the original shield design, and the new design results in a less conspicuous instrument that may be advantageous in many field ecology settings.

With the advent of small, low-cost environmental sensors, it is now possible to deploy high-density networks of sensors to measure hyper localized temperature variation. Here, we provide a detailed methodology for constructing a compact version of a previously described custom-fabricated radiation shield for use with inexpensive thermochrons.

一种用于空气温度传感器的紧凑型低成本辐射屏蔽的生态场研究

Low cost temperature sensors are increasingly used by ecologists to assess climatic variation and change on ecologically relevant scales. Although ...

Visualizing Efficacy of Pesticides Against Disease Vector Mosquitoes in the Field

Efficacy of public health pesticides targeting nuisance and disease-vector insects such as mosquitoes, sand flies, and filth-breeding flies is not uniform across ecological zones. To best protect public and veterinary health from these insects, the environmental limitations of pesticides need to be investigated to inform effective use of the most appropriate pesticide formulations and techniques. We have developed a research program to evaluate combinations of pesticides, pesticide application equipment, and application techniques in hot-arid desert, hot-humid tropical, warm and cool temperate, and urban locations to derive pesticide use guidelines specific to target insect and environment. To these ends we designed a system of protocols to support efficient, cost-effective, portable, and standardized evaluation of a diverse range of pesticides and equipment across multiple environments. At the core of these protocols is the use of an array of small cages with colony-reared sentinel mosquitoes (adults and immatures) and sand flies (adults), strategically arranged in natural habitats and exposed to pesticide spray. Spatial and temporal patterns of pesticide efficacy are derived from percent mortality in sentinel cages, then mapped and visualized in a geographic information system. Maps of sentinel mortality data may be statistically compared to evaluate relative efficacy of a pesticide across multiple environments, or to study multiple pesticides in a single environment. Protocols may be modified to accommodate a variety of scenarios, including, for example, the vertical orientation of sentinels in canopy habitats or simultaneous testing of ground and aerial application methods.

The efficacy of public health pesticides targeting nuisance and disease-vector insects is not uniform across different ecological zones. Here we present a system of techniques using captive vector insects as sentinels for pesticide efficacy to derive electronic maps supporting the standard evaluation of pesticides across multiple environments.

农药在田间对病媒蚊子的影响

Efficacy of public health pesticides targeting nuisance and disease-vector insects such as mosquitoes, sand flies, and filth-breeding flies is not uniform ...

Construction of a Low-cost Mobile Incubator for Field and Laboratory Use

Incubators are essential for a range of culture-based microbial methods, such as membrane filtration followed by cultivation for assessing drinking water quality. However, commercially available incubators are often costly, difficult to transport, not flexible in terms of volume, and/or poorly adapted to local field conditions where access to electricity is unreliable. The purpose of this study was to develop an adaptable, low-cost and transportable incubator that can be constructed using readily available components. The electronic core of the incubator was first developed. These components were then tested under a range of ambient temperature conditions (3.5 &#176;C - 39 &#176;C) using three types of incubator shells (polystyrene foam box, hard cooler box, and cardboard box covered with a survival blanket). The electronic core showed comparable performance to a standard laboratory incubator in terms of the time required to reach the set temperature, inner temperature stability and spatial dispersion, power consumption, and microbial growth. The incubator set-ups were also effective at moderate and low ambient temperatures (between 3.5 &#176;C and 27 &#176;C), and at high temperatures (39 &#176;C) when the incubator set temperature was higher. This incubator prototype is low-cost (&lt; 300 USD) and adaptable to a variety of materials and volumes. Its demountable structure makes it easy to transport. It can be used in both established laboratories with grid power or in remote settings powered by solar energy or a car battery. It is particularly useful as an equipment option for field laboratories in areas with limited access to resources for water quality monitoring.

This paper describes a method for building an adaptable, low-cost and transportable incubator for microbial testing of drinking water. Our design is based on widely available materials and can operate under a range of field conditions, while still offering the advantages of higher-end laboratory-based models.

建造供野外和实验室使用的低成本移动孵化器

Incubators are essential for a range of culture-based microbial methods, such as membrane filtration followed by cultivation for assessing drinking water ...

巴西田间放生转基因 伊 蚊的大规模生产

巴西田间放生转基因伊蚊的大规模生产