我々は、コンピュータグラフィックスのヘルプは昆虫館とナターシャSostromの援助のためにキースShigeteniに感謝。この作品は、USDA - ARSによって賄われていた。本書で述べられた意見、所見、結論または勧告は著者のものであり、必ずしも米国農務省の見解を反映するものではありません。 USDAは、機会均等のプロバイダや雇用主です。

1。寄生虫感染のためにホストのミバエの卵を準備します。 <ol><li>いっぱいに料理、サイドあたりの長さが10センチと1.5センチの深さ、寒天（よりコスト効果の高いオプションを代わりに、Gelcarin GP812、FMCバイオポリマー、ユーイングNJ）を準備することによって寄生さに、ショウジョウバエの卵のために基板を準備します。水1リットル当たり9グラムの濃度で寒天を準備します。 </li><li>液体培地（約70ミリリットル）とリムに料理をご記入のうえ、（少なくとも45分）、それらが冷却固化することができます。 </li><li>寒天ブロックが固体になれば、各ブロックの上部を覆って、ティッシュペーパーの単層（ジョージアパシフィック&quot;環境設定&quot;のバッチ組織の単層）を適用します。組織は、理由の表面の水分の寒天に容易に守らなければならない。 </li><li>寒天ブロックの組織覆われた表面に水に懸濁し、0.5mlの体積のミバエの卵を適用します。私たちの昆虫館では、これはBactrocera背 （ml当たり12000卵）の約6000の卵と同等です。 </li><li>幅広いブラシは組織の表面に均等にミバエの卵を広げるために水に浸した清潔な2.5センチメートルを使用してください。 </li><li>産卵を可能にするために寄生ケージの下画面の下のミバエの卵を置きます。卵の妨げになる死んだ寄生虫を除去するために底画面の下側をタップします。下の画面が卵に触れていない場合、それが最善です。 </li><li>寄生虫は、一晩と翌日に約21時間を産卵することができます。これは、寄生する卵の割合が高いのに十分な時間です​​。より長い曝露リスクのsuperparasitization。 </li></ol> 2。寄生宿主のショウジョウバエの孵化と蛹化<ol><li>面を上に卵とショウジョウバエの食事療法19の1.25リットル当たりの寄生卵の3から3.5準備寒天ブロックを配置することにより、コンテナの飼育準備。 </li><li>大きな蛹化容器に細かいグレードのバーミキュライトまたは洗浄砂の1.5奥行き層以上ライザーの卵でダイエットの容器を置きます。とマスキングテープの端蓋とテープで蛹化容器をカバーしています。バーミキュライトは、（下記参照）ふるい分け後に再利用できます。 </li><li>食事の容器と下vemiculiteに20を &quot;ポップ&quot;からクロールしに、ショウジョウバエの幼虫の蛹化容器の蓋の上部の間に十分なスペースがあることを確認してください。 </li><li> 27で部屋に蛹化容器を移動℃、一週間では80％の相対湿度（RH）。最初の4日間の暗い布でカバーのコンテナは、残りの日のための布を取り除く。 </li><li>一週間後に幼虫の食を削除、一度幼虫がポップされ、蛹化のためのバーミキュライトを入力している。バーミキュライトの幼虫がある場合、それらは一晩蛹になることができます。 </li><li>ふるいバーミキュライトからサナギ。粗いふるい（約3mm）で始まるは、バーミキュライトの残りの塊を除去するために手のふるいを使用してください。 </li></ol> 3。ソートと選択<ol><li>蛹含むF. arisanusは部分的にサイズがunparasitizedショウジョウバエから分離することができます。これらは寄生ハエの濃縮されるように、直径で1.65、2.26ミリメートルの間で蛹を収集する。主に小柄な寄生を含む、より大きなさなぎが主にハエが含まれているより小さいは12 pupariumの複数形は。寄生蛹のために女性の割合はサイズ12で正の相関があることに注意してください。 </li><li>様々な方法は、サイズがサナギをソートするために用いることができる。我々は、カスタムサイズソーター21を用いて機械的な方法を説明します。 </li><li>徐々にサイズ選別機を供給する振動フィーダーの上部に漏斗で蛹を置きます。 </li><li> 30 °の角度で少し発散圧延棒のペアで構成される大きさの選別、中に蛹を養う。漏斗を通してten別の容器に空のスロットの配列へのサイズによって個別にサナギのドロップ。 </li><li> 1.65〜2.26ミリメートルの大きさの範囲内のコップからサナギを収集し、ほとんどのハエがunparasitizedサナギから出てまで、さらに7日間出現ケージの中に置いてください。 </li><li>次に、それらはまだ含まれている昆虫から空のpupariumの複数形を区別するためにファンを使用してください。 </li></ol> 4。F. arisanusの出現とメンテナンス<ol><li> F.のコロニーを維持するためにarisanusは 、取り外し可能なガラスのフロントとサイドあたりの長さが約25cmの寄生保持するケージを使用してください。ケージは、1mm 2の上にスクリーニングし、側面の二つがあります。ケージの背面には、アクセス9 cmの穴とゴムシートで覆われている。ケージの底には、外寄生産卵するため、ショウジョウバエの卵を持つ2つの寒天ブロックの配置を可能にするために十分な大きさ1〜2ミリメートル2画面で内側に覆われてカットされています。 </li><li>保持しているケージはまた上昇率で、その結果、ケージの前面に沿って寄生混雑を作成することができる光を最小限に抑えるためにガラスの前面の底部四半期に沿って色を付けられた部分を持つ必要があります。 </li><li>これらの選択範囲の約11 gを置きますプラスチック容器にテッド蛹直径約9センチメートル。これはケージあたり600から700程度の寄生虫を生成する必要があります。サイズの選択は、約60％の女性になるはずです。 </li><li>コンテナへのスクリーン蓋を取り付けます。蓋は、成人F.できるようにする2ミリメートル2スクリーニング、で覆われるべきarsianus出現時にコンテナを残すことが、残りのミバエが含まれます。 </li><li> 24で寄生虫を維持° Cと12時12分日長と通気性のいいRH約45％。高い死亡率が観察されている場合は、1日に数時間または増加させる換気のために屋外にケージを移動すると、それを減らすことができます。寄生虫は、1つの追加週間で産卵する準備が整っているはずですし、さらに2週間のための生産的でなければなりません。 </li><li>ストリークは、ケージの上部に沿って週または使用可能な場合は蜂蜜が乾燥して少なくとも3回のスピン蜂蜜を原液。指先を使って幅の狭い縞で紡糸蜂蜜を適用します。縞が重すぎる場合は蜂蜜は、ケージに滴下することに注意してください。明るすぎる場合蜂蜜はすぐに乾燥して頻繁に再適用が必要になります。 </li><li>寄生虫に水分を提供するために、ケージの上部の上に置いて寒天ブロック（10センチ× 10センチメートル× 4cm）を。これらは、2回週を変更する必要があります。 </li></ol> 5。代表的な結果： ここでは、USDA - ARS F.に実施した品質管理手順の結果を報告する1月5日およ ​​び2010年6月22日の間にヒロ、ハワイでarisanus飼育操作。このビデオの記事とプロトコルで説明されているスケールでヒロの場所で飼育すると、2009年8月に開始され、そして新しい場所で最初の問題との調整は主に月までに解決されていた。したがって、これらのデータは、結果の代表的な他の研究者が説明したように彼らがプロトコルに従えば、そのコロニーを開始入手し、一般的に昆虫を飼育経験がある場合があります。この期間中、F.の小さなコロニーを維持していたarisanus：約3600 F.に相当する週当たりの生産4寄生蜂保有ケージ、 arisanus。 ホールディングケージ（幼虫食が削除された直後に、すなわちとの開発のさらに1週間前に2.26ミリメートル（濃縮） - 豊かな寄生虫感染率の最初のチェックは1.65の大きさの範囲内の蛹の2gのサンプルを取ることによって実施された蛹は、ふるいにかけたとサイズはソートさ。私たちは、&quot;初期のサナギ&quot;としてこれらを参照してください）​​。 2010年の最初の三ヶ月の間に濃縮されたサンプルの平均割合の寄生虫は0.46（SD = 0.18）であった。次の四半期中にこの平均値は、新しい場所での植民地と飼育手続の安定化を反映し、0.58（SD = 0.08）であった。が正常に寄生されなかった濃縮サナギがemergentsを生成しなかったものとハエを生産するものが含まれることに注意してください。 空のpupariumの複数形の保持ケージの取り外しでさらに1週間後に別の2グラムのサンプルが（&quot;後半蛹&#39;）取られた。ミバエは、この2番目のサンプルの1％未満の表現。ショウジョウバエの低い割合が以前の寄生虫と比較してハエの出現、その保持する容器に残ったほとんどのハエに部分的に原因です。 最後に、図1は、1月から6月2010サナギの質量の収率が得られる。利回りは、利息の大きさの範囲内で蛹濃縮、1.65〜2.26ミリメートルから測定されます。調整が新たな施設に対して行われていた間、今年の初めに&quot;早期蛹&quot;の大きな質量は、その時のミバエとunemerged蛹の高い割合を示している。 <img src="/files/ftp_upload/2901/2901fig1.jpg" alt="図1" /> 図1初期の濃縮（蛹の直径1.65〜2.26ミリメートル）の収率（すぐにサイズの選択後）とヒロ、ハワイでARS - USDA F. ariasnusの植民地での&quot;遅い&quot;（一週間のために保持し、空のケーシングを取り除いた後）サナギ、2010年1月〜6月

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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>

我々は報告書への関心の競合はありません。

このプロトコルとそれに付随する映像の記事ではF.を飼育するための最適化されたプロトコルを説明し、実証しているariasnus、実験室の設定でTephritidショウジョウバエの寄生蜂、。このプロトコルは、コロニーを維持するために必要な労働力と専門的な機器の量を最小限に抑えるために長年にわたって洗練されています。我々は、ホストのミバエの十分に確立、生産性と安定したコロニーは、寄生の飼育におけるすべての試みのために必要であることに注意してください。私たちの昆虫館では、ホストのミバエとしてBactrocera背を使用していますが 、他の種も同様に14から17有能なホストであることが示されている。 F.のいくつかの側面arisanusのコロニーの維持が検討されずに残っている。これらは、この種の10の研究室の適応の速度、22学習のメカニズムと適応とホストのミバエの可塑性に関与することができるメカニズムで発生する可能性のある遺伝的変化が含まれています。 フランス領ポリネシア、BにB.背：過去10年間、特に世界中Dacineのハエの蔓延の複数の例は、属Bactroceraのものが行われている南米の一部でcarambolae、B.アフリカとBのinvadensアフリカと地中海北岸でzonata。3,23,18は、F.などの生物的防除剤の有効性をテストする他のBactrocera種に対するarisanusは高い優先順位でなければ、そしてそれは、このプロトコルとそれに付随する映像資料に記載の方法のアプリケーションは、F.に関する研究を加速することを願っています場所24の多種多様でarisanus。最後に、出発点としてこの方法を用いてさらなる研究がまた新たな卵寄生25,1の植民地化に重要な情報を提供することがあります

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

Fopius arisanus（Sonan）は、少なくとも二つの理由Tephritidミバエの重要な寄生蜂である。最初に、それは卵のステージ1の間にホストに感染することが知られてわずか3 opiineの寄生の一つです。第二に、それは潜在的なショウジョウバエのホストの広い範囲を持っています。おそらく、その生活史、F.によるarisanusは正常に複数の熱帯地域で2-4のショウジョウバエの生物学的制御のために使用されています。 F.の広い使用には1つ障害ミバエの制御のためのarisanusは、安定した実験室コロニー5-9を確立することが困難なことです。この困難性にもかかわらず、1990年代に米国農務省の研究者は、F.の実験室個体群を維持するために信頼性の高い方法を開発arisanus 10-12。 F.に大きな関心があるarisanus生物学13,14、特にTephritidホスト14から17までの様々なコロニーを形成する能力については、関心は、特に過去10年間18の新たな大陸へBactroceraのミバエの害虫の驚くべき普及により駆動されます。 F.に関するさらなる研究arisanusと生物的防除剤としてのこの種の追加の展開では、飼育方法の最適化と改善の恩恵を受ける。このプロトコルと関連するビデオの記事ではF.を飼育するための最適化方法を説明します。 arisanusは、前述のアプローチ12に基づいて。我々はここで説明する方法により、F.の飼育世界中の、比較的低い肉体労働を必要とする熱帯地域で入手可能な材料を使って果物を使用せずに小規模でarisanus、。

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は正常にこれらの重要な熱帯の害虫の生物的防除に使用されているTephritidミバエの卵-幼虫寄生蜂である。ここではFを飼育するための最適化されたプロトコルを記述する容易に入手できる材料を使って小規模のarisanus。

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 ...

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Research

JoVE Journal

Biology

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

14.7K ビュー.  US Pacific Basin Agricultural Research Center. Fopius arisanusは正常にこれらの重要な熱帯の害虫の生物的防除に使用されているTephritidミバエの卵-幼虫寄生蜂である。ここではFを飼育するための最適化されたプロトコルを記述する容易に入手できる材料を使って小規模のarisanus。

ビデオ: 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

ショウジョウバエの勉強攻撃（ミバエ類）

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

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.

モスキート子育てのためのプロトコル（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 ...

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.

果実キューティクルの単離と生物物理学的研究

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.

電子鼻を使用した果実の揮発性の分析

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.

イメージングと免疫組織化学のための網膜ホールマウントの準備のために最適化されたプロトコル

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.

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 ...

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.

Simple Homemade Tools to Handle Fruit Flies&amp;#8212;&lt;em&gt;Drosophila melanogaster&lt;/em&gt;

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.

フルーツハエを扱うシンプルな自家製ツール -ショウジョウバエメラノガスター

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.

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. ...

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.

寄生性ガナスピス・ブラジリエンシスの飼育方法、侵襲性ショウジョウバエの有望な生物学的防除剤

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

無菌ナ ソニア スズメバチの飼育

Optimizing the Rearing Procedure of Germ-Free Wasps

Aseptic rearing technology is a method of culturing insects under sterile or almost sterile conditions, which can effectively eliminate the influence of external microorganisms on insect microbiota and thus promote the rapid development of insect microbiota research. Nasonia (wasp genus) is a parasitic wasp insect that has many advantages, such as a short lifespan, high genetic variation, easy operation, etc., and is widely used as an insect model system. Unlike antibiotic treatment, which can only reduce the number of microorganisms in animals, aseptic rearing techniques can control both the composition and quantity of microorganisms in animals, further facilitating the study of host-microbe interactions. However, previous versions of Nasonia rearing medium (NRM) have some defects and problems, such as a complex and time-consuming preparation process, easy contamination by bacteria or fungi, and short storage time. Therefore, this study solves these problems by optimizing the tools used in the NRM preparation process, storage conditions, and component ratios. The optimized medium could allow storage at -20 &#176;C for at least 3 months and eliminate the possibility of NRM contamination during feeding sterile wasps. This further improves the survival rate and health level of aseptic Nasonia, which is important for using Nasonia as a model for microbial research.

著者スポットライト:無菌スズメバチの飼育手順の最適化  

Nasonia wasp embryos were dissected from Lucillia sericata pupae after parasitization for 12-24 h and washed with alcohol and 10% sodium hypochlorite solution to obtain germ-free embryos. After rearing the germ-free embryos&#160;and supplying them with Nasonia rearing medium to grow and develop in vitro, germ-free Nasonia adults were obtained.

無菌スズメバチの飼育手順の最適化

Aseptic rearing technology is a method of culturing insects under sterile or almost sterile conditions, which can effectively eliminate the influence of ...