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

アンソニー Weinhold と私たちの昆虫飼育の手順を改善するための提案と同様、テクニカル サポートの年にわたって私たちと働いている多数の学生に感謝したいと思います。

1. 採卵 (図 1)
<ol><li>Egging カップを用意し、暖かい (~ 40 ° C) 水で満たされた ~ 500 mL ビーカーに布の片方の端を置きます。カップの両側をオーバー ラップし、輪ゴムで固定します。カップの上に戻って布の緩い端を折る。</li>
	<li>Egging カップ ケージ ~ 2 h. 妊娠女性の安定した古い 8-10 日ハエの飛ぶ場所が布のプレハブハウスします。</li>
	<li>スクイズ洗浄ボトルを小さな鍋に egging 布オフ ケージとリンスの卵から egging カップを削除します。</li>
</ol>2. 幼虫飼育のパンを準備します。
<ol><li>
メディア (1 飼育パンの量) の準備します。
	<ol>
<li>小麦ふすま (500 g)、木の削りくず (200 g) と魚の食事 (115 g) 10 L プラスチックの皿パンで組み合わせるし、よく混ぜます。</li>
		<li>(~ 25 ° C、1600 mL) の水に炭酸水素アンモニウム (50 g) を加え、溶けるまでを混ぜます。</li>
		<li>水ソリューションを乾燥した原料および unwetted 材料がないことを確認するためにミックスに追加します。パンの中のレベルが、圧縮しません。</li>
	</ol>
</li>
	<li>
安定した卵のフライを追加します。
	<ol>
<li>パンの長さ中の浅い溝を作る。</li>
		<li>卵ピペット (図 2) を溝に安定して飛ぶ卵 (卵 ~ 8,000) 1 mL を入金します。 注: 必要な時点で、100 を挿入するプラスチック卒業ピペットを切削で作られ卵ピペット メッシュ スクリーンと戻って一緒に接着のピペット。</li>
		<li>卵の乾燥を防ぐために培地の薄層をカバーしてください。</li>
		<li>カバー、ピローケース、パン、ラバー バンドとラベルを閉じます。幼虫の部屋で鍋を配置 (23 ± 2 ° C、30-50% RH 12:12 [L:D] h 日長)。 注: 幼虫の発育は、産卵の時から 10-14 日かかります。</li>
	</ol>
</li>
</ol>3. 蛹処理、幼虫は媒体の端へ移動 13 または 14 日産卵と Pupariate の後の 7-9 日
<ol><li>主にパンを培地の表面に形成し、きれいなフライパンに地殻の下の端のまわりにある puparia をスクープします。</li>
	<li>水の完全な 3/4 に ½ puparia で鍋を埋めます。Puparia 中小の塊を破る。培地の小麦ふすまとタンパク質成分シンク puparia ながら > 1 日古いフロート19。</li>
	<li>浮遊物、puparia、ザルやふるい (#5、7 #、#12 #20) 残りの媒体を削除する一連の洗浄といくつかの中のコンポーネントを削除します。#12 ふるい収集、puparia し #20 ドレインを入ることから固形廃棄物を保持します。シンク噴霧器とふるい間 puparia を洗います。</li>
	<li>#12 ふるいにきれいなフライパンに、puparia をすすぐ。水の ½ の完全にパンをご記入ください。ザルにフローティング puparia を注ぐでき、余分な水分を排出します。</li>
	<li>転送はファンと乾燥スクリーンに puparia を排水し、ドライ (図 3) まで残します。</li>
</ol>4. 蛹棚、Puparia を収集するための代替メソッドは、蛹の棚18
注: 棚は、10 L 料理鍋 (31.9 cm x 10.2 cm x 10.2 cm) の端からプラスチックのカットの部分から行われます。
<ol><li>2.1 で説明されるように媒体を準備します。テーパー中 ~7.5 cm ~2.5 cm 深さの 1 つの端からのパンの他に深い。</li>
	<li>
飼育のパンと幼虫が棚 (図 4) 横をクロールするを防ぐためにテープの浅い端の中に棚を配置します。
	<ol>
<li>スポンジ (~14.5 × 9.0 x 4.5 cm3、再生セルロース) を飽和水に浸した布 (30.5 x 42 cm2綿) や媒体からの約 1 つの半分インチの棚に場所で包む。</li>
	</ol>
</li>
	<li>
2.2 で説明されているように、卵を追加し、前に進みます。
	<ol>
<li>スポンジもしっとりしていることを毎日鍋を確認します。</li>
	</ol>
</li>
	<li>棚、スポンジ、きれいな皿に布からリンス puparia はパンおよび 14 日産卵後ザルに注ぐ。</li>
</ol>5. 品質管理
<ol><li>パン (総重量) で生産された puparia のすべての重量を量る。</li>
	<li>100 puparia を分離、重量を量る 9 cm シャーレに配置されます。</li>
	<li>カウントとセックス大人 〜 5-8 日後に出現率と性比を決定します。必要な場合、後日カウントするため冷凍庫に大人が登場し、シャーレを格納します。ハエが彼らの目の形と眼窩板20または低倍率下で性器の幅によって性欲を刺激します。</li>
	<li>レコードの総重量、重量 100 puparia、現れた大人の男性と女性の数。</li>
</ol>6. 血液の準備
<ol><li>70 g 水 500 mL にクエン酸ナトリウム第三水和物を含む 19 L バケツにローカルの屠畜場から新鮮な牛の血液を収集します。攪拌は、5-10 分は積極的に血液や研究室に戻った。</li>
	<li>削除するザルを通してひずみ血液凝固し、コレクションと冷凍庫 (-20 ° C) に保管の日付とラベルを 2 L 容器に分割します。</li>
	<li>冷凍庫、冷蔵庫内の場所から削除コンテナー 2 日前に必要です。〜 2 週間一度解凍し、1 年間冷凍保存血を使用できます。</li>
</ol>7. 成人のメンテナンス
<ol><li>ライン ケージ下部洗浄を容易にする肉屋の紙にケージを準備します。</li>
	<li>各ケージに 50 g の乾燥安定したハエ蛹 (~ 3,500) を配置します。大人 eclose 蛹処理後数日に数時間。</li>
	<li>場所新鮮な血液を供給するため各ケージの上に女性のナプキンを浸した。成虫の羽化の 24 h 内で給餌を開始し、産卵 (出現後 9-10 日) の前日まで毎日繰り返します。</li>
	<li>(セクション 1)、産卵後 4-8 時間冷凍庫にケージを置くし、紙を取り外し、frass の側面お湯で床からを洗浄および洗剤と塩素系漂白剤で表面を洗浄してきれい。お湯でケージを洗うし、乾燥することができます。</li>
</ol>

<ol>
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安定したハエは、さまざまな自然の中の基板の開発を見つけることができますや、研究室では、メディアの多くの種類で飼育することができます。充填剤として木の削りくずやバーミキュ ライトを使いました。バーミキュ ライトがうまく、しかし、飼育困難な媒体から分離 puparia 製高価 (~$0.60/pan)。おそらく puparia を培地から分離する追加の厳しさのためには、羽化もバーミキュ ライト、木...

安定したハエ、第(l.)、吸血性のハエが痛みを伴う刺され家畜の放牧行動を混乱させる、痛みを引き起こすとに苦しんでいる動物を仲間し世界人間のレクリエーション活動を中断します。発酵栄養物、しばしば動物の排泄物で汚染された未熟な安定したハエを開発します。農業慣行の変化と作物作物残渣、オーストラリア1の野菜、ブラジル2サトウキビとパイナップル コスタリカ3で安定したハエの深刻な流行を生み出した。動物あたりのちょうど 14 の安定したハエは経済のしきい値の4になることが、最近の発生5の中に動物あたりの以上 2,000 のハエの観測が行われました。このような寄生レベルはゼロに近いホスト生産性が低下し、死亡率6があります。農学関連の集団発生、結果として安定したハエは更新された興味を受け取っているし、研究所の植民地の需要が大幅に増加します。
すべての holometamorphic の昆虫に関しては安定したハエは未熟か幼虫の段階で成長に必要な栄養素のすべてを取得します。したがって、飼育システムの重要なコンポーネントは、幼虫のダイエットや基板です。フィールド7で基板の広い範囲の開発、安定したハエの幼虫を観察されているし、彼らは基板8,9の微生物のコミュニティに依存しています。天然幼虫基板は、分解または多くの場合窒素廃棄物で汚染された植物原料を発酵の主に構成されます。
研究所の飼育、安定したハエ幼虫基板、通常加えられた窒素源となる植物素材構成されます。多数の材料を安定して飛ぶ幼虫ダイエットのため使用されています。最初の幼虫ダイエット自然基板を真似た、エンバクわらと馬や牛の肥料10,11を発酵します。炭水化物源にはが含まれて小麦にはふすまには12 には,13,14, アルファルファの食事12,13,14と特殊化学品が開発した商用の処方製造業者協会 (csma 方式、33% 小麦ふすま、27% アルファルファの食事, 40% ビール酵母粒)13,14,15,16。酵母懸濁液12魚粉、炭酸水素アンモニウム17窒素源が含まれます。不活性材料の充てんは、エンバク殻12、バガス13、バーミキュ ライト16、木材チップ13,18小球形にされたピーナツの殻14などダイエットで含まれています。
飼育室の主な目的は、実験の結果がフィールドの人口のそれらを反映するために、生理学的、できるだけ「野生型」に似ている製品を生産するです。これは繁殖で、選択を最小化する必要があります遺伝的に維持するために多様性と栄養のリソース フィールドのそれらに匹敵します。副次目的は労働および費用を最小限に抑えるです。費用を最小限に抑えることの主要なコンポーネントは、ローカルで使用できる食事成分の。サシバエの飼育紹介システムは、これらの目標を満たすために開発されました。

<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

安定したハエ、第は、家畜、人間、伴侶動物および野生動物の世界の深刻な害虫です。最後の 20 + 年の間に農業慣行の変更は、いくつかの国で安定したハエの深刻な発生になった。これらの流行は、家畜の生産、この害虫の研究、管理作業を向上するための公開の要求の結果として人間のレクリエーションを中断しました。研究室の研究のための安定したハエを飼育のためのシンプルで安価なプロシージャが表示されます。プロシージャは、ローカルで使用できる食事成分、機器および消耗品を使用します。他有翅のハエの顔フライ (イエバエ autumnalis)、ノサシバエ (ノサシバエ製剤)、イエバエ (イエバエ) などを飼育の手順を適用できます。手順は、平均 12.5 mg と 〜 35% 卵成虫生存に安定して飛ぶ puparia を生成します。約 3000 のハエは、各パンの生産されています。

幼虫 pupariate 10-14 日間大人は 14-16 日産卵後出てくる。生成時間、卵には 〜 24 日です。図 5に 3 つの異なる増量エージェントと 2 人のコロニー 2013年 5 月 1 月 2017 年までの飼育データが掲載されています。コットンウッドを与えた最高の収量、3867 ±1442 (<img alt="Equation" src="/files/ftp_upload/57341/57341eq1.jpg"> </img> ± 標準偏差) 大人 2,872 ±1294 を生産研究所...

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

安定したハエ (第) を飼育する手順が表示されます。手順は、食事成分、機器および消耗品のローカルで利用可能な材料を使用します。

研究所は、安定したハエや他の有翅の双翅目の飼育

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

Universidad de Cartagena UDEC

Research

JoVE Journal

Biology

8.5K ビュー.  ARS-USDA. 安定したハエ (第) を飼育する手順が表示されます。手順は、食事成分、機器および消耗品のローカルで利用可能な材料を使用します。

ビデオ: 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.

子育てと研究のEristalis テナックスハナアブの長期メンテナンス

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

環境学

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.

再現可能な dsRNA のインジェクションと蚊とハエの産卵生物検定

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.

安価なセンサで昆虫の検出と分類をフライング

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

生物工学

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.

子育てに最適化されたプロトコル Fopius arisanus、Tephritid果実の寄生ハエ

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.

のメンテナンス<em&gt;キイロショウジョウバエ</em&gt;人口ケージ

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.

ショウジョウバエ・メラノガスターの概日伝播

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.

昆虫農業と無脊椎動物の生理学のための汎用性の高いオルソプテランモデルである Gryllus bimaculatusの実験室培養の維持

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.

蚊やショウジョウバエに対する殺虫剤毒性を定量するための局所適用バイオアッセイ

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.

下部双翅目バエ Bradysia(Sciara)coprophilaの実験室維持:新/古い新興モデル生物

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

ショウジョウバエのメンテナンスと飼育

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