Name: evaluating dryocosmus kuriphilus-induced damage on castanea sativa
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著者は、この研究を部分的に資金調達のための環境局ティチーノの森林サービスおよび中央政府のオフィスに感謝しています。

1. 木の選定・評価設計
<ol><li>可能であれば、信頼できる情報源 (例えば、科学を用いたクリタマバチとTorymus ススキとt. シネンシス寄生率の到着の年を決定することによって研究領域の流行段階を特定します。出版物、森林サービス、チェスナット グローブ マネージャーの知識)。<ol>
<li>信頼できる情報源が利用できない場合は、下記観測と組み合わせてt. チャ寄生率を計算することによって 4 つの主要な流行段階 (初期、ピーク、回復、生物防除) を識別します。<ol>
<li>樹冠も重大な損傷を示すとき初期の流行としてステージの識別もクラウンの透明性、現在の年苛立つはまれで、 t. チャ寄生が非常に低いまたは無視。</li>
			<li>樹冠表示、透明度の高い、枯れ枝はまれで、現在の年えいが豊富なときは、流行のピークを識別します。</li>
			<li>現在および以前の年のえいが豊富 (3 年前まで) とt. チャ寄生はまだ非常に低いまたは不在の長期にわたる流行のピークを識別します。樹冠はまだ透明度の高いを表示、追加ダメージは王冠の枯れ枝の最初の証拠で表されます。</li>
			<li>T. チャ寄生率は 7510より大きくなるときは、早期回復を識別します。被害は依然として高いが、現在の年の数の減少をガルズし、いくつかの枝生成特に頂芽からの胆嚢無料撮影。</li>
			<li>T. チャ寄生率は完全に 75% を超える、現在年えいが稀で、通常は単一の木だけに限定し、ほとんど枝生産胆嚢無料撮影するときは、回復期を識別します。過去の古い枝、枯れ枝D. クリタマバチのために年の虫こぶの攻撃がまだ表示されます。</li>
			<li>識別段階を完全に回復損傷や苛立つ (過去と現在の年) 非常に稀または不在、歯冠が完全に回復します。ひどく傷つけられた木、いくつかの名残で (死んだ撮影などや過去の年の苛立つ腐った) 事前D. クリタマバチの攻撃することができますまだクラウン内部に存在。 注: 補足ファイル 1 は、流行段階ごとに模範的な木の王冠の写真を示しています。</li>
		</ol>
</li>
	</ol>
</li>
	<li>ダメージ変動の中で、ツリー内を視覚的に推定する地域全体で栗の木を観察します。初期の流行と (クラウンは基本的に健全な) 回復段階の間にだけでなく、流行のピーク時にダメージ変動は通常低い (全体の冠は苛立つの満ちている)。対照的に、変動は過去のD. クリタマバチ攻撃のため死んだ撮影がまだ存在する場合高流行の中間段階でする傾向があります。</li>
	<li>1.1 と 1.2 に基づき、分析する木の数を決定します。残念ながら、それはないもフィールドや研究の目的で特定の流行状況に応じて異なる場合がありますサンプル サイズに関する特定のルールを与える適切です。以下お勧め私たちの 10 年の経験、10 ヘクタールの敷地のために基づいて (また詳細については表 1を参照)。<ol>
<li>流行の段階に関係なく、サイトごとに少なくとも 10 の木をサンプルします。回復期および早い流行 3 本の木は、十分だろうが、10 にサンプル サイズを増やしてより統計的に力を与える結果。</li>
		<li>回復期および初期の流行、ツリーごとの 1 つの分岐をサンプルします。</li>
		<li>流行のピーク時に樹冠内均等な苛立つかより深刻ないくつかの王冠の部分を攻撃されていることに気づく場合ツリーごと 2 つのブランチ ツリーごとの 1 つの分岐をサンプルします。</li>
		<li>他の伝染性の段階で (よく回復している木) 2 つまたは 3 つに枝の数を増やす (より多くの損傷木) すべての木の王冠のダメージの変動に基づきます。</li>
	</ol>
</li>
</ol>フィールドのデータ コレクション
<ol><li>クリップボード、キャンプ椅子、剪定はさみ、伸縮式の木の剪定、30 m 測定テープおよびツリー クライミング装置 8 m 以上トップ クラウン解析が必要な場合、適切な機器を準備します。</li>
	<li>それに比例して樹冠内支店の多様性をカバーしようと最も代表的な分岐を選択 (健全木通常支店を持っているような破損した木が枝の損傷の程度が異なるとあるかもしれませんが)。たとえば、ツリーごとの 3 つの枝を収集する場合は、被害が最も支店、健康と、中間的なものを収集します。</li>
	<li>可能な限り、reiterations (トランク吸盤または reiterations扇子Hallé)16を避けながら建築枝のみを選択します。</li>
	<li>枝は、長さ 50 cm 以上、少なくとも 10 の撮影があることを確認します。</li>
	<li>切削点で枝の地面の上の高さを測定するために伸縮ツリー剪定はさみの刃に近い測定テープの初めを接続します。</li>
	<li>伸縮式剪定はさみで枝を切って、その刈取り高さの視点は、その型を記録 (建築または繰り返し)、分析の部分だけを維持するために剪定はさみとブランチの選択を絞り込みます。</li>
	<li>ブランチに一意の ID を割り当てるし、その時代の一般的な情報について (頂点に最初の分岐ポイント) から、最大の長さを記録します。</li>
	<li>その歴史と現状 (重く攻撃か) の最初の印象を取得する全支店で簡単に見て、すべての要素および図 2と図 3の助けを借りて、DCI の計算の重要な機能を示します。</li>
</ol>3. 分岐機能の定義
次の定義は、· ゲーリングらからで部分的または完全に再現します。20185シュプリンガー社ベルリン ハイデルベルク 2017 の権限を持つ。
<ol><li>撮影に発展芽から現在の生長の季節の間に生えている形成されたばかりの芽として、もやし(から撮影) を定義します。</li>
	<li>サンプリング日に関して、前生長する季節からスプラウトとして撮影を定義 (例えば、採取時期 = 2017、撮影 = 2016 年に育ったスプラウト)。デッドオアア ライブをすることができます。</li>
	<li>D. クリタマバチ攻撃後または自然死のため死んだ撮影として死んで撮影(Sd) を定義します。</li>
	<li>再アクティブ化された休眠芽と混同しないように、生きている撮影、ライブ撮影(として) を定義します。</li>
	<li>以上の撮影は、多年にわたる分岐部に休眠芽から現在の生長の季節の間に生えている形成されたばかりの芽として再度有効に休眠芽(Bdor) を定義します。</li>
	<li>胆嚢の基部や芽の軸に沿って開発としての撮影で胆嚢(Gon) を定義します。技術的には、これらが「もやしに虫こぶ"と呼ばれる必要があります既存の文献との整合性、目的のため我々 は「撮影に苛立つ」としてそれらを参照。 注:図 2と図 3は、選択された分岐の機能の例を示します。詳細かつ完全な説明 (このホワイト ペーパーの範囲を超えています) · ゲーリングらであります。20185と Maltoniら20124。</li>
</ol>4. 分岐解析
<ol><li>数とレコードすべての生活 (生きている撮影) の撮影します。</li>
	<li>カウントし、死んだすべてのシュートを記録します。</li>
	<li>カウントし、再アクティブ化されたすべての休眠芽を記録します。</li>
	<li>カウントし、撮影にすべての虫こぶを記録します。 注: 補足のファイル 2 フィールド サンプリング フォームの例を示します、補足ファイル 3 記入サンプル フォームを示しています。</li>
</ol>5. 損傷複合インデックスの計算
<ol><li>死んでシュート シュート (死んだ撮影 + 生きている撮影) の合計数で割った値の数から死んだ枝 (Sd) の割合を計算します。</li>
	<li>再アクティブ化された休眠芽生稈 (BdoR + 生きている撮影) の合計数で割った値の数から再アクティブ化された休眠芽 (BdoR) の比率を計算します。</li>
	<li>撮影生活撮影 (BdoR + 生きている撮影) の数で割った値に虫こぶの数から撮影 (Gon) に虫こぶの数の平均値を計算します。</li>
	<li>DCI の DCI の数式を使用して計算 = (Sd * 0.479 + BdoR * 0.525 + Gon * 0.120) * 100。</li>
	<li>損傷の重症度を評価するため、表 2を使用します。 注: DCI 関数 R スクリプト補足符号化ファイル 1 で利用可能です。補足的なファイル 4 にその説明があります。</li>
</ol>

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著者が明らかに何もありません。

クリタマバチは、春に苛立つの形成を誘導する栗の木芽の卵を産みます。繰り返されると制御不能はD. クリタマバチは攻撃形成、光合成の葉エリア5で多くの撮影および重大な損失の死を含む一般的な分岐の破損だけでなく、原因です。木は通常休眠芽の活性化を介して代替撮影を生成しようとして反応します。このため、流行のピーク時と回復期、 D. クリ?...

栗 gallwaspクリタマバチ安松 (ハチ目: に関する)クリ1,2,3属の最も重要な世界的な害虫です。その繰り返しのかじり活動を通じての予防し、通常撮影開発4,5, 葉面積の削減の進行やツリー グリーン バイオマスと活力5,6 の結果としての損失の原因を抑制します。、休眠芽の活性化5と gallwasp ポスト出現支店死亡7、8に増加。
Gallwasp 流行の欧州の経験は、制御されていないと繰り返し gallwasp 攻撃が甘い栗 (ヨーロッパグリミル) クラウン汚職の高レベルを引き起こす可能性がありますを示しています。これは、休眠芽の活性化も同じ植生期間5中 2 番目のフラッシュを構築することにより生成される代替葉によってどちらも補償は、70% までの冠葉区域の損失につながります。
害虫の人口を減らすし、栗の木を回復する唯一の成功のメソッドはその自然な拮抗薬Torymus ススキ上條寄生蜂による生物的防除 (ハチ目: 大陸)9,10。その天敵を生物学的コントロールが達成されれば、栗の木は新しい健康な芽を生成する開始します。ツリー損傷レベルが非常に高い、この可能性がありますのみ、冬芽から始まって gallwasp 産卵活動4後その形成のため通常のインフェ ステーション無料だという事実のために。これは、全体の樹冠が再確立5前に長い回復プロセスを意味します。
後Torymus ススキによる生物的防除に達し、その林業 (剪定、間伐) の介入の必要性を確認するには、森林管理者と栗の栽培方法が必要に迅速な栗の木の陽性反応を確認するために、gallwasp 流行の害虫による初期の侵入段階から回復中の損傷レベルと関連ブランチ アーキテクチャと葉地域進化その拮抗薬による生物的防除後信頼性の高い評価。Gallwasp 侵入度 (メイド) を評価するための手法を開発し、攻撃の芽11またはえい芽12あたりの平均数の割合を測定などの日付に世界中で使用します。グリーン バイ オ マス (例えば、葉面積)、休眠芽など準備構造、反応構造 (休眠芽など再アクティブ化と 2 番目のフラッシュ)、または前年、メイドを測定しない直接メジャーとして被害 (例えば、死んだ撮影)現在ツリー活力と気力6,13,14のプロキシ。さらに、ほとんどのメイドはのみ苛立つペスト流行 (図 1) のピーク時に特に木の枝と過小評価本物の枝損傷発見の数に基づいています。
本稿で述べる · ゲーリングらによって提案された複合インデックス (DCI) アプローチ損傷グリーン バイ オ マスのプロキシを考慮した 2018年5留保休眠芽など流行のすべての段階を通じて被害の現実的な, 信頼性の高い、かつ合理的に迅速な評価を有効にするツリー反応 (休眠芽再活性化や第二フラッシュ)· ゲーリングらによって提案された評価努力最適化と組み合わせる場合は特に201715。
本稿の目的は 1) 2) 損傷の複合指数を提示する、3)、改善された重症度スケール変換を提案するなどと評価される関連するブランチ機能を含むフィールド プロトコルの詳細な説明を与えるため、in particular、します。DCI。

<table><tbody><tr><td>Name of Material/ Equipment</td><td>Company</td><td>Catalog Number</td><td>Comments</td></tr><tr><td>Clipboard</td><td>Any brand</td><td></td><td></td></tr><tr><td>Camping chair&lt;br/&gt; (Foldable and lightweight chair)</td><td>Any brand</td><td></td><td>Companies: Kelty, Campz, Half-Ton.</td></tr><tr><td>Felco 9 secateurs&lt;br/&gt; (One-hand pruning shear)</td><td>Felco</td><td></td><td>Other companies: Bahco.</td></tr><tr><td>AP-5M-Aluminium Pole&lt;br/&gt; (Telescopic tree pruner pole)</td><td>Bahco</td><td>8152079</td><td>Other companies: Spear &amp;amp; Jackson, Kingfisher, Hortex, Fiskars.</td></tr><tr><td>P34-37 top pruner&lt;br/&gt; (Telescopic tree pruner head)</td><td>Bahco</td><td>8002787</td><td></td></tr><tr><td>100 ft Fiberglass Long Tape&lt;br/&gt; (30 m measuring tape)</td><td>Stanley</td><td>34-790</td><td>Other companies: Tjima, Freemans, Astor, Lux.</td></tr><tr><td>Parallel 10.5mm&lt;br/&gt; (Low stretch kernmantel rope, flexible and lightweight for rope access)</td><td>Petzl</td><td>R077AA03</td><td>Basic equipment for tree climbing&amp;nbsp; (if necessary). Many other equipment configurations can be used for tree climbing, depending on the situation and on single operator preferences. We used Pezl equipment but many other companies offer similar products (e.g. Edelrid, Notch, Climbing technologies, DMM, ...). For a complete overview of equipment and companies we recommend a search in google&amp;nbsp; &quot;tree climbing gear&quot; as search keyword. PLEASE NOTE: tree climbing activities should be done only by professionals and are submitted to specific regulatory prescriptions according to the country.</td></tr><tr><td>Avao Sit&lt;br/&gt; (Harness for work positioning and suspension)</td><td>Petzl</td><td>C69AFA 2</td><td></td></tr><tr><td>Rig&lt;br/&gt; (Compact self-braking descender)</td><td>Petzl</td><td>D21A</td><td></td></tr><tr><td>Ascension&lt;br/&gt; (Handled rope clamp for rope ascents)</td><td>Petzl</td><td>B17ALA</td><td></td></tr><tr><td>Eclipse&lt;br/&gt; (Storage for throw-line)</td><td>Petzl</td><td>S03Y</td><td></td></tr><tr><td>Airline&lt;br/&gt; (Throw-line)</td><td>Petzl</td><td>R02Y 060</td><td></td></tr><tr><td>Jet&lt;br/&gt; (Throw-bag)</td><td>Petzl</td><td>S02Y 300</td><td></td></tr><tr><td>Vertex best&lt;br/&gt; (Comfortable helmet for work at height and rescue)</td><td>Petzl</td><td>A10BYA</td><td></td></tr><tr><td>Zillon&lt;br/&gt; (Adjustable work positioning lanyard for tree care)</td><td>Petzl</td><td>L22A 040</td><td></td></tr><tr><td>Ok&lt;br/&gt; (Lightweight oval carabiner)</td><td>Petzl</td><td>M33A SL</td><td></td></tr></tbody></table>

evaluating dryocosmus kuriphilus-induced damage on castanea sativa

クリタマバチ安松は、ヨーロッパに到着以来、 Castaneasativaの主要な害虫になっています。そのかじり活動別ゴール型の形成に結果し、通常のシュートの開発を防ぐことができます。苛立つと葉面積のアテンダント胆嚢低下の生産、休眠芽の活性化における分岐パーツ、および増加の死を含むブランチ アーキテクチャの進歩的な破損以外にも、繰り返されると制御不能の攻撃を引き起こします。これまでのところ、支店建築被害を定量化するためのほとんどの試みが します。さらに、侵入度 (メイド) を評価するためさまざまな方法されている開発苛立つの存在と豊かさにのみ焦点を当てる。
辺材面積関係に葉面積を使用すると、緑色のバイオマスの指標として、我々 開発以前の研究で最も重要な支店の建築的特徴を考慮した損傷複合インデックス (DCI) の現実的な損傷評価を可能にします。プロセス全体の流行。
本研究の目的は、この手法を提示し、他の広く使用されている指標に関して破損の説明の違いを強調することです。結果を示す DCI で良く、枝損傷を描いたメイドと比較して、流行のピーク時に特にそれを過小評価する傾向が.我々 は古典的なメソッド、およびクラウン透明性評価を使用して侵入度複合損傷インデックスによる病害虫の全体への影響を正しく評価する方法を示唆することによって締結します。

全胆嚢ハチ流行段階を網羅時空間勾配を作成するために 2013、2016 年までの間、スイスのティチーノ州に 25 の地域の合計に訪問されました。合計では、我々 を収集および流行初期 (害虫とツリーの破損の最初の到着) で 5 つのサイトで 94 枝、流行のピーク ( D. クリタマバチ攻撃の高レベルのための深刻な被害の中) で 5 つのサイトで 200 本の枝を分析、回復期 ( <...

Watch this Scientific Journal Video about Evaluating Dryocosmus Kuriphilus-induced Damage on Castanea Sativa at JoVE.com

Evaluating Dryocosmus Kuriphilus-induced Damage on Castanea Sativa

クリタマバチの評価-ヨーロッパグリを照射

苛立つだけの豊かさを考慮したのではなく、また考慮に関連事業所破損を取って、クリタマバチの被害を評価するの一般的です。従ってより現実的な損傷の評価を有効にする最も重要な分岐機能を考慮した複合損傷指標を提案する.

Dryocosmus kuriphilus Yasumatsu has become a major pest for Castaneasativa since its arrival in Europe. Its galling activity results in the formation of ...

evaluating-dryocosmus-kuriphilus-induced-damage-on-castanea-sativa

Research

JoVE Journal

Environment

Evaluating Dryocosmus Kuriphilus-induced Damage on Castanea Sativa

7.2K ビュー.  WSL Swiss Federal Research Institute. 苛立つだけの豊かさを考慮したのではなく、また考慮に関連事業所破損を取って、クリタマバチの被害を評価するの一般的です。従ってより現実的な損傷の評価を有効にする最も重要な分岐機能を考慮した複合損傷指標を提案する.

ビデオ: Evaluating Dryocosmus Kuriphilus-induced Damage on Castanea Sativa

A Technique to Screen American Beech for Resistance to the Beech Scale Insect (Cryptococcus fagisuga Lind.)

Beech bark disease (BBD) results in high levels of initial mortality, leaving behind survivor trees that are greatly weakened and deformed.&#160;The disease is initiated by feeding activities of the invasive beech scale insect, Cryptococcus fagisuga, which creates entry points for infection by one of the Neonectria species of fungus.&#160;Without scale infestation, there is little opportunity for fungal infection. Using scale eggs to artificially infest healthy trees in heavily BBD impacted stands demonstrated that these trees were resistant to the scale insect portion of the disease complex1.&#160;Here we present a protocol that we have developed, based on the artificial infestation technique by Houston2, which can be used to screen for scale-resistant trees in the field and in smaller potted seedlings and grafts.&#160;The identification of scale-resistant trees is an important component of management of BBD through tree improvement programs and silvicultural manipulation.

A Technique to Screen American Beech for Resistance to the Beech Scale Insect Cryptococcus fagisuga Lind.

Beech bark disease is initiated by feeding activities of the beech scale insect that create fungal entry points in the bark.&#160;Trees that are resistant to the scale insect are also disease resistant.&#160;Here we present the protocol we have developed to screen individual beech trees for beech scale resistance.

ブナスケール昆虫への抵抗のためのアメリカのブナを（画面ための技術<em&gt;クリプトコッカスfagisuga</em&gt;リンド。）

Beech bark disease (BBD) results in high levels of initial mortality, leaving behind survivor trees that are greatly weakened and deformed.&#160;The ...

環境学

Remote Sensing Evaluation of Two-spotted Spider Mite Damage on Greenhouse Cotton

The objective of this study was to evaluate a ground-based multispectral optical sensor as a remote sensing tool to assess foliar damage caused by the two-spotted spider mite (TSSM), Tetranychus urticae Koch, on greenhouse grown cotton. TSSM is a polyphagous pest which occurs on a variety of field and horticultural crops. It often becomes an early season pest of cotton in damaging proportions as opposed to being a late season innocuous pest in the mid-southern United States. Evaluation of acaricides is important for maintaining the efficacy of and preventing resistance to the currently available arsenal of chemicals and newly developed control agents. Enumeration of spider mites for efficacy evaluations is laborious and time consuming. Therefore, subjective visual damage rating is commonly used to assess density of spider mites. The NDVI (Normalized Difference Vegetation Index) is the most widely used statistic to describe the spectral reflectance characteristics of vegetation canopy to assess plant stress and health consequent to spider mite infestations. Results demonstrated that a multispectral optical sensor is an effective tool in distinguishing varying levels of infestation caused by T. urticae on early season cotton. This remote sensing technique may be used in lieu of a visual rating to evaluate insecticide treatments.

This manuscript describes a multispectral optical sensor that effectively detected damage to early season cotton artificially infested with varying densities of the two-spotted spider mite populations.

温室効果コットンの二斑点スパイダーダニの被害のリモートセンシング評価

The objective of this study was to evaluate a ground-based multispectral optical sensor as a remote sensing tool to assess foliar damage caused by the ...

In Vitro Rearing of Solitary Bees: A Tool for Assessing Larval Risk Factors

Although solitary bees provide crucial pollination services for wild and managed crops, this species-rich group has been largely overlooked in pesticide regulation studies. The risk of exposure to fungicide residues is likely to be especially high if the spray occurs on, or near host plants while the bees are collecting pollen to provision their nests. For species of Osmia that consume pollen from a select group of plants (oligolecty), the inability to use pollen from non-host plants can increase their risk factor for fungicide-related toxicity. This manuscript describes protocols used to successfully rear oligolectic mason bees, Osmia ribifloris sensu lato, from egg to prepupal stage within cell culture plates under standardized laboratory conditions. The in vitro-reared bees are subsequently used to investigate the effects of fungicide exposure and pollen source on bee fitness. Based on a 2 &#215; 2 fully crossed factorial design, the experiment examines the main and interactive effects of fungicide exposure and pollen source on larval fitness, quantified by prepupal biomass, larval developmental time, and survivorship. A major advantage of this technique is that using in vitro-reared bees reduces natural background variability and allows the simultaneous manipulation of multiple experimental parameters. The described protocol presents a versatile tool for hypotheses testing involving the suite of factors affecting bee health. For conservation efforts to be met with significant, lasting success, such insights into the complex interplay of physiological and environmental factors driving bee declines will prove to be critical.

In Vitro Rearing of Solitary Bees: A Tool for Assessing Larval Risk Factors

Fungicide sprays on flowering plants may expose solitary bees to high concentrations of pollen-borne fungicide residues. Using laboratory-based experiments involving in vitro-reared bee larvae, this study investigates the interactive effects of consuming fungicide-treated pollen derived from host and non-host plants.

体外孤独な蜂の飼育: 幼虫の危険因子を評価するためのツール

Although solitary bees provide crucial pollination services for wild and managed crops, this species-rich group has been largely overlooked in pesticide ...

Ecotoxicological Methodologies to Evaluate Biomarkers at Different Scales in Neotropical Anurans

The new questions in ecotoxicology highlight the importance of applying a battery of biomarkers, as this results in ecotoxicological predictions that improve not only the interpretation of the effects of environmental stressors on organisms but also the determination of their possible impact. It is well known that the use of ecotoxicological biomarkers at different levels of organization allows for the prediction of the biological responses of organisms to environmental stressors, which is useful in environmental risk assessment. 
Nevertheless, it is necessary to consider the optimization of basic procedures, to generate historical data in control groups, and to employ specific bioassays to evaluate responses in organs and tissues in order to elucidate the nature and variation of the effects observed. Therefore, the present work aims to describe several ecotoxicological methodologies employed in all stages of neotropical anurans at different ecological levels and to validate them as useful biomarkers to be used both in wildlife and in laboratory conditions. In this work, these biomarkers were applied at the individual/organismic level (body condition index), histological/physiological level (histopathology, histometric, and pigmentary analyses), biochemical level (oxidative stress enzymes), and genetic level (direct and oxidative damage in DNA by comet assay). 
Although these methodologies have small variations or modifications depending on the species, these techniques provide effective biomarkers for evaluating the effect of xenobiotics on anurans, which possess certain characteristics that make them useful indicator species of aquatic and terrestrial ecosystems. In conclusion, the battery of biomarkers employed in the present study has proven to be adequate for estimating toxic responses in Neotropical anurans and can be further recommended as bioindicators for identifying the impact of pollutants on the aquatic ecosystems of the region. Finally, it is recommended to achieve the standardization of these important biomarkers for anurans in specific regions as well as to possibly include them in risk assessments and decision-making.

<meta charset="utf8"></head><body>新熱帯性無尿類におけるさまざまなスケールでのバイオマーカーを評価するための生態毒性学的方法論

In this paper, standardized ecotoxicological methods for the evaluation of biomarkers in neotropical anuran species are presented. Specifically, this paper details several methodologies at different scales of ecotoxicological evaluation, such as the genetic, cellular-histological, biochemical, morphological, and individual levels.

新熱帯性無尿類におけるさまざまなスケールでのバイオマーカーを評価するための生態毒性学的方法論

The new questions in ecotoxicology highlight the importance of applying a battery of biomarkers, as this results in ecotoxicological predictions that ...

Establishment of a Robust and Reproducible Model of Radiation-Induced Skin and Muscle Fibrosis

Radiation-induced skin fibrosis (RISF) can result from a plethora of scenarios including cancer therapy, accidental exposure, or acts of terrorism. Radioactive beams can penetrate through the skin and affect the structures in their path including skin, muscles, and internal organs. Skin is the first structure to get exposed to radiation and is susceptible to develop chronic fibrosis, which is challenging to treat. Currently, limited treatment options show moderate efficacy in mitigating radiation-related skin fibrosis. A key factor hindering the development of effective countermeasures is the absence of a convenient and robust model that could allow for translation of the experimental findings to humans. Here, a robust and reproducible murine hind limb skin fibrosis model has been established for prophylactic and therapeutic evaluation of possible agents for functional and molecular recovery.
The right hind limb was irradiated using a single dose of 40 (Gray) Gy to induce skin fibrosis. Subjects developed edema and dermatitis in the early stages proceeded by visible skin constriction. Irradiated limbs showed a significantly reduced limb range of motion in the following weeks. In late stages, acute side effects subsided, yet chronic fibrosis persisted. A gait index was performed as an additional functional assay, which demonstrated the development of functional impairment. These non-invasive methods demonstrated reliable measurements for tracing fibrosis progression, which is supported by histological analyses. The radiation dose, application, and post-irradiation analyses employed in this model offer a vigorous and reproducible method for studying radiation-induced skin fibrosis and testing the efficacy of therapeutical agents.

Here we present a protocol to induce radiation-induced skin fibrosis in the hind limb of mice and perform post-irradiation measurements of chronic impairment via limb excursion and gait index analyses to evaluate the functional outcome. The model elucidates radiation-related skin fibrosis mechanisms and is useful in subclinical therapeutic studies.

Radiation-induced skin fibrosis (RISF) can result from a plethora of scenarios including cancer therapy, accidental exposure, or acts of terrorism. ...

がん研究

Measuring Changes in Tactile Sensitivity in the Hind Paw of Mice Using an Electronic von Frey Apparatus

Measuring inflammation-induced changes in thresholds of hind paw withdrawal from mechanical pressure is a useful technique to assess changes in pain perception in rodents. Withdrawal thresholds can be measured first at baseline and then following drug, venom, injury, allergen,&#160;or otherwise evoked inflammation by applying an accurate force on very specific areas of the skin. An electronic von Frey apparatus allows precise assessment of mouse hind paw withdrawal thresholds that are not limited by the available filament sizes in contrast to classical von Frey measurements. The ease and rapidity of measurements allow for incorporation of assessment of tactile sensitivity outcomes in diverse models of rapid-onset inflammatory and neuropathic pain as multiple measurements can be taken within a short time period. Experimental measurements for individual rodent subjects can be internally controlled against individual baseline responses and exclusion criteria easily established to standardize baseline responses within and across experimental groups. Thus, measurements using an electronic von Frey apparatus represent a useful modification of the well-established classical von Frey filament-based assays for rodent mechanical allodynia that may also be applied to other nonhuman mammalian models.

Electronic von Frey measurements are an effective and improved alternative to classical von Frey filaments, as they allow rapid and precise measurement of changes in rodent mechanical withdrawal thresholds to continuously applied pressure stimuli before and after an inflammatory event.

Measuring inflammation-induced changes in thresholds of hind paw withdrawal from mechanical pressure is a useful technique to assess changes in pain ...

神経科学

An Improved Assay and Tools for Measuring Mechanical Nociception in Drosophila Larvae

Published assays for mechanical nociception in Drosophila have led to variable assessments of behavior. Here, we fabricated, for use with Drosophila larvae, customized metal nickel-titanium alloy (nitinol) filaments. These mechanical probes are similar to the von Frey filaments used in vertebrates to measure mechanical nociception. Here, we demonstrate how to make and calibrate these mechanical probes and how to generate a full behavioral dose-response from subthreshold (innocuous or non-noxious range) to suprathreshold (low to high noxious range) stimuli. To demonstrate the utility of the probes, we investigated tissue damage-induced hypersensitivity in Drosophila larvae. Mechanical allodynia (hypersensitivity to a normally innocuous mechanical stimulus) and hyperalgesia (exaggerated responsiveness to a noxious mechanical stimulus) have not yet been established in Drosophila larvae. Using mechanical probes that are normally innocuous or probes that typically elicit an aversive behavior, we found that Drosophila larvae develop mechanical hypersensitization (both allodynia and hyperalgesia) after tissue damage. Thus, the mechanical probes and assay that we illustrate here will likely be important tools to dissect the fundamental molecular/genetic mechanisms of mechanical hypersensitivity.

An Improved Assay and Tools for Measuring Mechanical Nociception in Drosophila Larvae

The goal of this protocol is to show how to perform an improved assay for mechanical nociception in Drosophila larvae. We use the assay here to demonstrate that mechanical hypersensitivity (allodynia and hyperalgesia) exists in Drosophila larvae.

ショウジョウバエ幼虫における機械的な結知を測定するための改善されたアッセイとツール

Published assays for mechanical nociception in Drosophila have led to variable assessments of behavior. Here, we fabricated, for use with Drosophila ...

行動学

Using Terminal Transferase-mediated dUTP Nick End-labelling (TUNEL) and Caspase 3/7 Assays to Measure Epidermal Cell Death in Frogs with Chytridiomycosis

Amphibians are experiencing a great loss in biodiversity globally and one of the major causes is the infectious disease chytridiomycosis. This disease is caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd), which infects and disrupts frog epidermis; however, pathological changes have not been explicitly characterized. Apoptosis (programmed cell death) can be used by pathogens to damage host tissue, but can also be a host mechanism of disease resistance for pathogen removal. In this study, we quantify epidermal cell death of infected and uninfected animals using two different assays: terminal transferase-mediated dUTP nick end-labelling (TUNEL), and caspase 3/7. Using ventral, dorsal, and thigh skin tissue in the TUNEL assay, we observe cell death in the epidermal cells in situ of clinically infected animals and compare cell death with uninfected animals using fluorescent microscopy. In order to determine how apoptosis levels in the epidermis change over the course of infection we remove toe-tip samples fortnightly over an 8-week period, and use a caspase 3/7 assay with extracted proteins to quantify activity within the samples. We then correlate caspase 3/7 activity with infection load. The TUNEL assay is useful for localization of cell death in situ, but is expensive and time intensive per sample. The caspase 3/7 assay is efficient for large sample sizes and time course experiments. However, because frog toe tip biopsies are small there is limited extract available for sample standardization via protein quantification methods, such as the Bradford assay. Therefore, we suggest estimating skin surface area through photographic analysis of toe biopsies to avoid consuming extracts during sample standardization.

Using Terminal Transferase-mediated dUTP Nick End-labelling TUNEL and Caspase 3/7 Assays to Measure Epidermal Cell Death in Frogs with Chytridiomycosis

We quantify epidermal cell death in frogs with chytridiomycosis using two methods. First, we use terminal transferase-mediated dUTP nick end-labelling (TUNEL) in situ histology to determine differences between clinically infected and uninfected animals. Second, we conduct a time series analysis of apoptosis over infection using a caspase 3/7 protein analysis.

ターミナル媒介 dUTP を使用してニック終わりラベル (TUNEL) とカスパーゼ 3/7 がカエルツボカビ、カエルのメジャー表皮細胞死に試金します。

Amphibians are experiencing a great loss in biodiversity globally and one of the major causes is the infectious disease chytridiomycosis. This disease is ...

免疫・感染学

Electroantennographic Bioassay as a Screening Tool for Host Plant Volatiles

Plant volatiles play an important role in plant-insect interactions. Herbivorous insects use plant volatiles, known as kairomones, to locate their host plant.1,2 When a host plant is an important agronomic commodity feeding damage by insect pests can inflict serious economic losses to growers. Accordingly, kairomones can be used as attractants to lure or confuse these insects and, thus, offer an environmentally friendly alternative to pesticides for insect control.3 Unfortunately, plants can emit a vast number volatiles with varying compositions and ratios of emissions dependent upon the phenology of the commodity or the time of day. This makes identification of biologically active components or blends of volatile components an arduous process. To help identify the bioactive components of host plant volatile emissions we employ the laboratory-based screening bioassay electroantennography (EAG). EAG is an effective tool to evaluate and record electrophysiologically the olfactory responses of an insect via their antennal receptors. The EAG screening process can help reduce the number of volatiles tested to identify promising bioactive components. However, EAG bioassays only provide information about activation of receptors. It does not provide information about the type of insect behavior the compound elicits; which could be as an attractant, repellent or other type of behavioral response. Volatiles eliciting a significant response by EAG, relative to an appropriate positive control, are typically taken on to further testing of behavioral responses of the insect pest. The experimental design presented will detail the methodology employed to screen almond-based host plant volatiles4,5 by measurement of the electrophysiological antennal responses of an adult insect pest navel orangeworm (Amyelois transitella) to single components and simple blends of components via EAG bioassay. The method utilizes two excised antennae placed across a "fork" electrode holder. The protocol demonstrated here presents a rapid, high-throughput standardized method for screening volatiles. Each volatile is at a set, constant amount as to standardize the stimulus level and thus allow antennal responses to be indicative of the relative chemoreceptivity. The negative control helps eliminate the electrophysiological response to both residual solvent and mechanical force of the puff. The positive control (in this instance acetophenone) is a single compound that has elicited a consistent response from male and female navel orangeworm (NOW) moth. An additional semiochemical standard that provides consistent response and is used for bioassay studies with the male NOW moth is (Z,Z)-11,13-hexdecadienal, an aldehyde component from the female-produced sex pheromone.6-8

A method to rapidly screen host plant volatiles by measurement of the electrophysiological response of adult navel orangeworm (Amyelois transitella) antennae to single components and blends via electroantennographic analysis is demonstrated.

宿主植物の揮発性物質のスクリーニングツールとしてElectroantennographicバイオアッセイ

Plant volatiles play an important role in plant-insect interactions. Herbivorous insects use plant volatiles, known as kairomones, to locate their host ...

クリタマバチの評価-ヨーロッパグリを照射

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ブナスケール昆虫への抵抗のためのアメリカのブナを（画面ための技術

温室効果コットンの二斑点スパイダーダニの被害のリモートセンシング評価

体外孤独な蜂の飼育: 幼虫の危険因子を評価するためのツール

新熱帯性無尿類におけるさまざまなスケールでのバイオマーカーを評価するための生態毒性学的方法論

新熱帯性無尿類におけるさまざまなスケールでのバイオマーカーを評価するための生態毒性学的方法論

Establishment of a Robust and Reproducible Model of Radiation-Induced Skin and Muscle Fibrosis

Measuring Changes in Tactile Sensitivity in the Hind Paw of Mice Using an Electronic von Frey Apparatus

ショウジョウバエ幼虫における機械的な結知を測定するための改善されたアッセイとツール

ターミナル媒介 dUTP を使用してニック終わりラベル (TUNEL) とカスパーゼ 3/7 がカエルツボカビ、カエルのメジャー表皮細胞死に試金します。

宿主植物の揮発性物質のスクリーニングツールとしてElectroantennographicバイオアッセイ