Name: laboratory rearing of stable flies and other muscoid diptera
Uploaded: 2018-08-03T12:30:00.000+00:00
Duration: 7 min
Description: Watch this Scientific Journal Video about Laboratory Rearing of Stable Flies and Other Muscoid Diptera at JoVE.com

我们要感谢安东尼. Weinhold 和多年来与我们合作的众多学生的技术支持, 以及改进我们的昆虫饲养程序的建议。

1. 收集卵子 (图 1)
<ol><li>准备好怂恿杯, 把布的一端放在500毫升的烧杯里, 里面装满了温暖的 (~ 40 °c) 的水。重叠的杯子的两侧, 并修复与橡胶带。将布料松散的一端折回杯顶。</li>
	<li>将怂恿杯放在8–10天的老苍蝇笼中 2 h. 妊娠雌性稳定的苍蝇会在布料上排卵。</li>
	<li>从笼中取出怂恿杯, 将怂恿布上的鸡蛋冲洗成一个小平底锅, 用挤压洗瓶。</li>
</ol>2. 准备幼虫饲养盘
<ol><li>
准备培养基 (1 饲养盘的数量)
	<ol>
<li>将麦麸 (500 克)、木屑 (200 克) 和鱼粉 (115 克) 组合在10升塑料碟盘中, 混合均匀。</li>
		<li>加入碳酸氢铵 (50 克) 到水 (~ 25 °c, 1600 毫升) 和混合, 直到溶解。</li>
		<li>添加水溶液的干燥配料和混合, 以确保没有 unwetted 材料。在平底锅中的水平介质, 但不压缩。</li>
	</ol>
</li>
	<li>
添加稳定的飞卵
	<ol>
<li>在中等长度的平底锅上作浅沟。</li>
		<li>将1毫升稳定的飞卵 (8000 卵) 在沟中用蛋吸管 (图 2) 存入。 注: 通过在所需点切割一个塑料刻度吸管, 插入100网格屏幕, 再将吸管粘在一起, 制成的蛋吸管。</li>
		<li>用一层薄薄的培养基盖上鸡蛋以防止脱水。</li>
		<li>盖平底锅与枕头盒, 关闭与橡胶带和标签。在幼虫室放置平底锅 (23 2 °c, 30–50% RH 和 12:12 [L: D] h 光周期)。 注意: 幼虫发育需要从产卵时10–14天。</li>
	</ol>
</li>
</ol>3. 蛹处理后, 幼虫在产卵和 Pupariate 13 天或14后移到培养基7–9天的边缘。
<ol><li>铲蛹, 主要位于锅的边缘周围的外壳, 形成在表面上的介质, 出和放置在一个干净的平底锅。</li>
	<li>装满平底锅, 用蛹, ½¾满水。分解蛹和培养基的团簇。蛹 > 1 天老浮19, 中间水槽的麦麸和蛋白质组分。</li>
	<li>去除浮动材料, 蛹和一些中等组分与漏勺并且洗涤通过一系列筛子 (#5, #7, #12, #20) 去除剩余的媒介。#12 筛收集蛹, #20 使固体废弃物进入排水管。用水槽喷雾器在筛子之间洗蛹。</li>
	<li>冲洗蛹, 在 #12 筛, 成一个干净的锅。把锅装满水½。将漂浮的蛹倒入漏勺中, 允许多余的水排出。</li>
	<li>将抽干的蛹放到烘干屏上, 直到干燥 (图 3)。</li>
</ol>4. 蛹货架, 另一种收集蛹的方法是与蛹货架18
注: 货架是由一个从10升碟盘 (10.2 厘米高 x 10.2 厘米宽 x 31.9 厘米长) 的一端的塑料切割。
<ol><li>准备介质如2.1 所述。锥形介质在平底锅从2.5 厘米深的一端到7.5 厘米深。</li>
	<li>
将架子放在饲养盘的浅端和胶带上, 以防幼虫爬行在架子上 (图 4)。
	<ol>
<li>饱和海绵 (~ 14.5 x 9.0 x 4.5 cm3, 再生纤维素) 并且包装它在水被浸泡的布料 (30.5 x 42 cm2, 棉花) 和地方在架子大约一个半英寸从媒介。</li>
	</ol>
</li>
	<li>
添加鸡蛋, 如2.2 所述, 并继续像以前一样。
	<ol>
<li>每天检查平底锅以确保海绵保持湿润。</li>
	</ol>
</li>
	<li>将蛹从货架上冲洗出来, 海绵和布料放入一个干净的盘子里, 产卵后14天倒入漏勺。</li>
</ol>5. 质量控制
<ol><li>称量所有在平底锅中生产的蛹 (总重量)。</li>
	<li>隔离100蛹, 称重并放置在9厘米培养皿中。</li>
	<li>伯爵和性成人 ~ 5–8天后确定出现率和性别比例。储存培养皿与出现的成年人在一个冰箱, 以计数在以后的日期, 如果需要的话。苍蝇是奸的形状, 他们的眼睛和宽度的额轨道板20或生殖器在低放大倍数。</li>
	<li>记录总重量, 重量100蛹, 并且出现成年男性和女性的数量。</li>
</ol>6. 准备血液
<ol><li>从当地屠宰场收集新鲜牛血19升桶, 含70克柠檬酸钠三元二水合物500毫升水。大力搅拌血液5–10分钟, 返回实验室。</li>
	<li>通过漏勺的血液来去除血块并分成2升的容器, 标签与收集的日期和储存在一个冰箱 (-20 °c)。</li>
	<li>在需要前2天, 从冰箱中取出容器并放入冰箱。血液可用于2周, 一旦解冻, 并储存在冰箱里长达1年。</li>
</ol>7. 成人保养
<ol><li>准备笼子, 线笼底部用屠夫纸方便清洗。</li>
	<li>在每个笼子里放置50克干燥稳定的飞蛹 (~ 3500)。成人 eclose 几个小时后, 蛹处理后几天。</li>
	<li>在每个笼子的顶部放一个新鲜的血液浸泡女性餐巾喂养。开始喂养24小时内成人羽化和重复每天直到产卵前的一天 (9–10天后出现)。</li>
	<li>产卵后 (1 节), 把笼子放在冰箱里 4–8 h, 然后清除纸张, 用热水冲洗两侧和地板上的取食量, 用洗衣粉和氯漂白剂擦洗所有表面。用热水冲洗笼子, 让干燥。</li>
</ol>

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作者没有什么可透露的。

稳定的苍蝇可以发现发展在广泛的各种基质的性质, 可以饲养在许多类型的媒体在实验室。我们用木屑和蛭石作膨胀剂。蛭石工作良好, 但使分离蛹从饲养培养基困难和昂贵 ($ 0.60/平底锅)。可能是由于增加了从培养基中分离蛹的严谨性, 羽化也降低了蛭石, 57% vs 75% 用于木屑。棉刨花可媲美蛭石, 但有时难以采购。最重要的成分似乎是植物或纤维素材料发酵和维持一个 "开放" 或良好的充气培养基和?...

稳定的苍蝇, Stomoxys calcitrans (L), 是 hematophagous 苍蝇, 其痛苦的叮咬扰乱牲畜的放牧行为, 造成痛苦和痛苦的同伴动物和扰乱人类娱乐活动的世界各地。不成熟的稳定蝇生长在发酵的植物物质, 经常被动物粪便污染。改变农艺做法和农作物造成了作物残留物的稳定蝇的严重暴发, 澳大利亚的蔬菜1, 巴西的甘蔗2, 哥斯达黎加的菠萝3。虽然每只动物的14只稳定苍蝇被认为是经济阈值4, 但在最近的疫情中, 每只动物的2000多只苍蝇的观测结果已经达到5。这种侵扰程度将主机生产率降低到接近零, 并可能导致死亡率6。由于 agronomically 相关的暴发, 稳定的苍蝇正在接受新的兴趣和对实验室殖民地的需求急剧增加。
对于所有 holometamorphic 昆虫, 稳定的苍蝇在未成熟或幼体阶段获得生长所需的所有养分。因此, 饲养系统的一个重要组成部分是幼虫饮食或基质。稳定的蝇幼虫已经观察到在田间7的广泛基质中发展, 它们依赖于基底8,9的微生物群落。天然幼虫基质主要由分解或发酵经常被含氮废物污染的营养物质组成。
在实验室饲养中, 稳定的蝇幼虫基质通常由营养物质和添加的氮源组成。大量的材料已用于稳定的飞行幼虫饮食。第一种幼虫饮食模仿天然基质, 包括发酵燕麦秸秆和马或牛粪10,11。碳水化合物来源包括麦麸12、13、14、苜蓿膳食12、13、14和由化学专业开发的商业配方。制造商协会 (CSMA, 33% 麦麸, 27% 苜蓿膳食, 40% 啤酒酵母颗粒)13,14,15,16。氮源包括酵母悬浮液12、鱼粉和碳酸氢铵17。惰性膨胀材料通常包括燕麦壳12, 蔗渣13, 蛭石16, 木片13,18和颗粒花生壳14。
实验室饲养的一个主要目标是生产一种与 "野生型" 相似的产品, 以便实验室实验能产生反映田间人群的结果。这就要求最大限度地减少育种和选择, 以维持遗传多样性, 并使营养资源与实地的情况相媲美。次要目标是尽量减少劳动和开支。减少开支的一个主要组成部分是利用当地现有的饮食成分。为实现这些目标, 研制了稳定的飞养系统。

<table><tbody><tr><td>Diamalt</td><td>Premier Malt Products, Inc., Saddle Brook, NJ</td><td>2540</td><td></td></tr><tr><td>CSMA Fly media</td><td>Purina Animal Nutrition, Arden Hills, MN</td><td>5S6Z</td><td></td></tr><tr><td>Thin Maxi Pad</td><td>The Tranzonic Co., Cleveland, OH, USA</td><td>5001M</td><td></td></tr><tr><td>Calf Manna</td><td>MannaPro, Chesterfield, MO, USA</td><td>Manna Pro</td><td></td></tr><tr><td>Ammonium Bicarbonate</td><td>Spectrum Chemical Manufacturing Corp, Gardena, CA</td><td>A1125</td><td></td></tr><tr><td>Wheat bran, Coarse</td><td>Siemer Milling Company, Teutopolis, IL</td><td></td><td></td></tr><tr><td>Wood shavings</td><td>Tractor Supply Company, Brentwood, TN</td><td>502770699</td><td></td></tr><tr><td>Fishmeal</td><td>Consumer Supply Distributing, North Sioux City, SD</td><td>F1550</td><td></td></tr><tr><td>Adult cages</td><td>All Aluminum Window Company, Lincoln, Ne</td><td>Custom</td><td>45 &amp;times; 45 &amp;times; 45 cm, 18 &amp;times; 16 mesh aluminum screen, stockinette access</td></tr><tr><td>9 &amp;times; 28 cm black cotton cloth</td><td>Robert Kaufman Fabrics, Los Angeles, CA</td><td>K040-114</td><td>Egging cloth</td></tr><tr><td>10 L plastic dish pans</td><td>Rubbermaid, Saratoga Springs, NY</td><td>FG2951ARWHT</td><td>Larval pans</td></tr><tr><td>Stockinette, Cotton, 12 inch x 25 yard roll</td><td>Tex-Care Medical Company, Burlington, NC</td><td>91311-225</td><td></td></tr></tbody></table>

laboratory rearing of stable flies and other muscoid diptera

稳定的苍蝇, Stomoxys calcitrans, 是家畜, 人类, 同伴动物和野生动物的严重害虫。在过去的20年中, 农艺做法的变化导致几个国家的稳定蝇的严重暴发。这些暴发扰乱了牲畜生产和人类的娱乐活动, 导致公众要求为这种害虫增加研究和管理工作。提出了一种简单、廉价的用于实验室研究的稳定蝇的饲养方法。该程序使用当地现有的饮食成分, 设备和用品。该程序可适用于饲养其他 muscoid 苍蝇, 包括面部苍蝇 (蝇 autumnalis), 喇叭飞行 (Haematobia irritans), 和房子苍蝇 (家蝇)。该程序生产稳定的飞行蛹平均12.5 毫克和35% 鸡蛋的成年生存。大约3000只苍蝇在每个平底锅被生产。

幼虫 pupariate 10–14天和成人在产卵后14–16天出现。世代时间, 蛋到蛋, 是 ~ 24 天。2013年5月至2017年1月的饲养数据有三种不同的膨胀剂和两个菌落,如图 5所示。白杨提供了最好的产量, 3867 ±1442<img alt="Equation" src="/files/ftp_upload/57341/57341eq1.jpg"> </img> (标准偏差) 蛹重12.5 ±1.6 毫克74±19% 羽化生产2872±1294成年蝇为建立的实验室殖民地。性别比例略有偏向女性 0...

Watch this Scientific Journal Video about Laboratory Rearing of Stable Flies and Other Muscoid Diptera at JoVE.com

Laboratory Rearing of Stable Flies and Other Muscoid Diptera

提出了一种饲养稳定蝇 (Stomoxys calcitrans) 的方法。该程序使用当地现有的材料, 为膳食成分, 设备和用品。

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 ...

laboratory-rearing-of-stable-flies-and-other-muscoid-diptera

Research

JoVE Journal

Biology

8.5K 次观看.  ARS-USDA. 提出了一种饲养稳定蝇 (Stomoxys calcitrans) 的方法。该程序使用当地现有的材料, 为膳食成分, 设备和用品。

视频: Laboratory Rearing of Stable Flies and Other Muscoid Diptera

Rearing and Long-Term Maintenance of Eristalis tenax Hoverflies for Research Studies

With an estimated 6000 species worldwide, hoverflies are ecologically important as alternative pollinators to domesticated honeybees. However, they are also a useful scientific model to study motion vision and flight dynamics in a controlled laboratory setting. As the larvae develop in organically polluted water, they are useful models for investigating investment in microbial immunity. While large scale commercial breeding for agriculture already occurs, there are no standardized protocols for maintaining captive populations for scientific studies. This is important as commercial captive breeding programs focusing on mass output during peak pollination periods may fail to provide a population that is consistent, stable and robust throughout the year, as is often needed for other research purposes. Therefore, a method to establish, maintain and refresh a captive research population is required. Here, we describe the utilization of an artificial hibernation cycle, in addition to the nutritional and housing requirements, for long term maintenance of Eristalis tenax. Using these methods, we have significantly increased the health and longevity of captive populations of E. tenax compared to previous reports. We furthermore discuss small scale rearing methods and options for optimizing yields and manipulating population demographics.

Rearing and Long-Term Maintenance of Eristalis tenax Hoverflies for Research Studies

The overall goal of these procedures is to establish, maintain and refresh a captive population of Eristalis tenax in a research setting.

研究性学习的Eristalis 管Hoverflies 的饲养和长期维护

With an estimated 6000 species worldwide, hoverflies are ecologically important as alternative pollinators to domesticated honeybees. However, they are ...

科研

环境科学

Reproducible dsRNA Microinjection and Oviposition Bioassay in Mosquitoes and House Flies

Synthetic dsRNAs, used to induce RNA interference, may have dose dependent phenotypic effects. These effects are difficult to define if the dsRNAs are delivered using a non-quantitative method. Accurate delivery of known quantities of nucleic acids or other chemicals is critical to measure the efficacy of the compound being tested and to allow reliable comparison between compounds.
Here we provide a reproducible, quantitative microinjection protocol that ensures accurate delivery of specific doses of dsRNA, reducing the mortality typically induced by injection injury. These modifications include the addition of Rhodamine B, a graduated injection needle, and an improved recovery method borrowed from Isoe and Collins. This method allows calculation of dose responses and facilitates comparisons between compounds. Versions of this method have been successfully used on three genera of mosquitoes as well as house flies to assess the reduction in fecundity resulting from gene silencing of ribosomal RNA transcripts.
This protocol provides strategies to reduce several challenges of small insect microinjection. Together, mechanical delivery of dsRNAs accompanied by visual verification, identification of effective locations for delivery, and inclusion of a post-injection recovery period ensure accurate dosing and low injury mortality. This protocol also describes an oviposition bioassay for uniform determination of effects on fecundity.

This protocol describes a microinjection methodology that we have standardized and used for several years to deliver specific quantities of nucleic acids directly to the hemolymph of mosquitoes and house flies. This protocol results in minimal injection mortality and allows dose correlated measurements of fecundity.

可重复的 dsrna 微注射和蚊子和家蝇的奥维恩生物测定

Synthetic dsRNAs, used to induce RNA interference, may have dose dependent phenotypic effects. These effects are difficult to define if the dsRNAs are ...

Flying Insect Detection and Classification with Inexpensive Sensors

An inexpensive, noninvasive system that could accurately classify flying insects would have important implications for entomological research, and allow for the development of many useful applications in vector and pest control for both medical and agricultural entomology. Given this, the last sixty years have seen many research efforts devoted to this task. To date, however, none of this research has had a lasting impact. In this work, we show that pseudo-acoustic optical sensors can produce superior data; that additional features, both intrinsic and extrinsic to the insect&#8217;s flight behavior, can be exploited to improve insect classification; that a Bayesian classification approach allows to efficiently learn classification models that are very robust to over-fitting, and a general classification framework allows to easily incorporate arbitrary number of features. We demonstrate the findings with large-scale experiments that dwarf all previous works combined, as measured by the number of insects and the number of species considered.

We proposed a system that uses inexpensive, noninvasive pseudo-acoustic optical sensors to automatically and accurately detect, count, and classify flying insects based on their flying sound.

飞行昆虫检测与分类用便宜的传感器

An inexpensive, noninvasive system that could accurately classify flying insects would have important implications for entomological research, and allow ...

生物工程

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

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 parasitoids known to infect the host during the egg stage1. Second, it has a wide range of potential fruit fly hosts. Perhaps due to its life history, F. arisanus has been a successfully used for biological control of fruit flies in multiple tropical regions2-4. One impediment to the wide use of F. arisanus for fruit fly control is that it is difficult to establish a stable laboratory colony5-9. Despite this difficulty, in the 1990s USDA researchers developed a reliable method to maintain laboratory populations of F. arisanus10-12. There is significant interest in F. arisanus biology13,14, especially regarding its ability to colonize a wide variety of Tephritid hosts14-17; interest is especially driven by the alarming spread of Bactrocera fruit fly pests to new continents in the last decade18. Further research on F. arisanus and additional deployments of this species as a biological control agent will benefit from optimizations and improvements of rearing methods. In this protocol and associated video article we describe an optimized method for rearing F. arisanus based on a previously described approach12. The method we describe here allows rearing of F. arisanus in a small scale without the use of fruit, using materials available in tropical regions around the world and with relatively low manual labor requirements.

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

Fopius arisanus is an egg-larval parasitoid of Tephritid fruit flies that is successfully used in biological control of these important tropical pests. We describe here an optimized protocol for rearing F. arisanus on a small scale using readily available materials.

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 ...

生物学

Maintenance of a Drosophila melanogaster Population Cage

Large quantities of DNA, RNA, proteins and other cellular components are often required for biochemistry and molecular biology experiments. The short life cycle of Drosophila enables collection of large quantities of material from embryos, larvae, pupae and adult flies, in a synchronized way, at a low economic cost. A major strategy for propagating large numbers of flies is the use of a fly population cage. This useful and common tool in the Drososphila community is an efficient way to regularly produce milligrams to tens of grams of embryos, depending on uniformity of developmental stage desired. While a population cage can be time consuming to set up, maintaining a cage over months takes much less time and enables rapid collection of biological material in a short period. This paper describes a detailed and flexible protocol for the maintenance of a Drosophila melanogaster population cage, starting with 1.5 g of harvested material from the previous cycle.

Maintenance of a Drosophila melanogaster Population Cage

This manuscript reports a detailed protocol for culturing, on a regular basis, a population of Drosophila melanogaster using a fly population cage.

的维持<em&gt;果蝇</em&gt;人口笼

Large quantities of DNA, RNA, proteins and other cellular components are often required for biochemistry and molecular biology experiments. The short life ...

Circadian Entrainment of Drosophila Melanogaster

Nearly universal among organisms, circadian rhythms coordinate biological activity to earth&#39;s orbit around the sun. To identify factors creating this rhythm and to understand the resulting outputs, entrainment of model organisms to defined circadian time-points is required. Here we detail a procedure to entrain many Drosophila to a defined circadian rhythm. Furthermore, we detail post-entrainment steps to prepare samples for immunofluorescence, nucleic acid, or protein extraction-based analysis.

Here, we detail how to synchronize Drosophila to a circadian day. This is the first, and most important step necessary for studying biological rhythms and chronobiology.

果蝇黑色素的循环训练

Nearly universal among organisms, circadian rhythms coordinate biological activity to earth&#39;s orbit around the sun. To identify factors creating this ...

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

Gryllus bimaculatus (De Geer)&#160;is a large-bodied cricket distributed throughout Africa and Southern Eurasia where it is often wild-harvested as human food. Outside its native range, culturing G. bimaculatus is feasible due to its dietary plasticity, rapid reproductive cycle, lack of diapause requirement, tolerance for high-density rearing, and robustness against pathogens. Thus, G. bimaculatus can be&#160;a versatile model for studies of insect physiology, behavior, embryology, or genetics.
Cultural parameters, such as stocking density, within-cage refugia, photoperiod, temperature, relative humidity, and diet, all impact cricket growth, behavior, and gene expression and should be standardized. In the burgeoning literature on farming insects for human consumption, these crickets are frequently employed to evaluate candidate feed admixtures derived from crop residues, food-processing byproducts, and other low-cost waste streams.
To support ongoing experiments evaluating G. bimaculatus growth performance and nutritional quality in response to variable feed substrates, a comprehensive set of standard protocols for breeding, upkeep, handling, measurement, and euthanasia in the laboratory was developed and is presented here. An industry-standard cricket feed has proven nutritionally adequate and functionally appropriate for the long-term maintenance of cricket breeding stocks, as well as for use as an experimental control feed. Rearing these crickets at a density of 0.005 crickets/cm3 in screen-topped 29.3 L polyethylene cages at an average temperature of 27 &#176;C on a 12 light (L)/12 dark (D) photoperiod, with moistened coconut coir serving both as hydration source and oviposition medium has successfully sustained healthy crickets over a 2-year span. Following these methods, crickets in a controlled experiment yielded an average mass of 0.724 g 0.190 g at harvest, with 89% survivorship and 68.2% sexual maturation between stocking (22 days) and harvest (65 days).

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

This paper outlines basic methods to standardize important factors such as density, feed availability, hydration source, and environmental controls for the long-term rearing of laboratory cultures of the edible cricket, Gryllus bimaculatus.

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

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

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. ...

Bradysia coprophila Lab Protocols and Chromosomal Research

Laboratory Maintenance of the Lower Dipteran Fly Bradysia (Sciara) coprophila: A New/Old Emerging Model Organism

Laboratory stocks of the lower dipteran fly, Bradysia (Sciara) coprophila, have been maintained for over a century. Protocols for laboratory upkeep of B. coprophila are presented here. These protocols will be useful for the rapidly increasing number of laboratories studying B. coprophila to take advantage of its unique biological features, which include (1) a monopolar spindle in male meiosis I; (2) non-disjunction of the X dyad in male meiosis II; (3) chromosome imprinting to distinguish maternal from paternal homologs; (4) germ line-limited (L) chromosomes; (5) chromosome elimination (paternal chromosomes in male meiosis I; one to two X chromosomes in early embryos; L chromosomes from the soma in early embryos); (6) sex determination by the mother (there is no Y chromosome); and (7) developmentally regulated DNA amplification at the DNA puff loci in larval salivary gland polytene chromosomes. 
It is now possible to explore these many unique features of chromosome mechanics by using the recent advances in sequencing and assembly of the B. coprophila genome and the development of transformation methodology for genomic engineering. The growing scientific community that uses B. coprophila for research will benefit from the protocols described here for mating the flies (phenotypic markers for mothers that will have only sons or only daughters; details of mass mating for biochemical experiments), checking embryo hatch, feeding larvae, and other comments on its rearing.

Author Spotlight: Exploring Bradysia coprophila's Unique Biology &#8211; A Guide to Laboratory Maintenance

This paper outlines the laboratory maintenance (including mating and feeding) of the lower dipteran fly Bradysia (Sciara) coprophila.

下双翅目蝇 Bradysia （Sciara） coprophila 的实验室维护:一种新的/旧的新兴模式生物

Laboratory stocks of the lower dipteran fly, Bradysia (Sciara) coprophila, have been maintained for over a century. Protocols for laboratory upkeep of B. ...

Drosophila Maintenance

Drosophila melanogaster, commonly known as fruit flies, are a frequently used model organism for life science research. Although starting a collection of these critters may seem as easy as leaving a banana on your kitchen counter for too long, a productive fly colony in the lab requires careful husbandry and maintenance.
This video demonstrates the necessary steps for maintaining a healthy fly stock. The overview begins with the preparation and storage of the yeast and sugar-containing media on which flies feed. Next, the vessels most commonly used for housing Drosophila are shown, as well as how and when to move flies between these containers. Finally, the presentation also includes examples of the ways in which housing and feeding conditions are manipulated for biological experiments.

Maintenance and Husbandry of Drosophila melanogaster

果蝇的饲养与保存

Drosophila melanogaster, commonly known as fruit flies, are a frequently used model organism for life science research. Although starting a collection of ...

稳定蝇及其他 Muscoid 双翅目的实验室饲养

摘要

探索更多视频

此视频中的章节

研究性学习的Eristalis 管Hoverflies 的饲养和长期维护

可重复的 dsrna 微注射和蚊子和家蝇的奥维恩生物测定

飞行昆虫检测与分类用便宜的传感器

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

的维持

果蝇黑色素的循环训练

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

局部应用生物测定以量化蚊子和果蝇的杀虫剂毒性

下双翅目蝇 Bradysia （Sciara） coprophila 的实验室维护:一种新的/旧的新兴模式生物

果蝇的饲养与保存

稳定蝇及其他 Muscoid 双翅目的实验室饲养

摘要

探索更多视频

此视频中的章节

研究性学习的Eristalis 管Hoverflies 的饲养和长期维护

可重复的 dsrna 微注射和蚊子和家蝇的奥维恩生物测定

飞行昆虫检测与分类用便宜的传感器

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

的维持

果蝇黑色素的循环训练

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

局部应用生物测定以量化蚊子和果蝇的杀虫剂毒性

下双翅目蝇 Bradysia （Sciara） coprophila 的实验室维护:一种新的/旧的新兴模式生物

果蝇的饲养与保存

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