この作品は、熱帯 AgriSciences 部の内部補助金機関によって支えられた (プロジェクト番号: 20175013) と両方の付与、チェコの大学生命科学からたばこ 20182023。

1. 肉準備
<ol><li>地元の肉屋から牛 (大腿二頭筋から新鮮な牛) のロースを入手して研究室に転送します。 注: 気密密閉袋に入れて 20 分より長くない期間常温 (20-25 ° C)、牛ロース肉を輸送する勧めします。</li>
	<li>層流の安全キャビネットで、牛筋の外側の表面を殺菌する 10 の 70% (v/v) エタノールを噴霧して筋肉を洗うスクイズ ボトル 500 mL を使用して s。筋表面の 1 cm2あたりエタノールの 0.025 g を適用します。</li>
	<li>筋肉の内側に残りのエタノールを避けるためにナイフで肉の外側の表面を無菌削除します。筋肉の表面の均一性を保つために筋肉の内側の約 3 mm を削除します。</li>
	<li>プラスチック製の密封された袋の筋をパッケージ化し、冷凍庫に転送すること。</li>
	<li>1 日-18 ° C で筋肉を格納します。その後、6 h 4 ° C で凍結筋を解凍します。 注: 融解、勧め筋肉を冷凍庫から冷蔵庫に移動します。</li>
	<li>層流安全キャビネットで各筋肉カッターで 0.5 cm 厚切りにスライスします。ナイフで切ると小さな 5 × 2.5 cm2長方形のサンプル。</li>
	<li>パッケージのビニール袋に長方形の肉サンプル、-18 ° c 保存用冷凍庫で保管します。</li>
</ol>2. 標準化と層流安全キャビネット内接種手順の準備
<ol><li>標準化された接種 (1.5 × 108 CFU/mL)エシェリヒア属大腸菌ATCC 25922 肉サンプルを接種するを準備します。<ol>
<li>ストックの接種材料の準備のため最初入力済みに 10 ml の滅菌バッファー ミュラー ヒントン スープ (BMHB) 15 mL 滅菌チューブに (製造業者によって配信) 凍結乾燥細菌培養を調剤します。37 ° C で 24 時間この懸濁液を育成します。<ol>
<li>次のように細菌の貯蔵液の準備: 取る約 0.1 - 0.2 mL の細菌懸濁液 20 mL バイアルに希薄で閉じてゴム栓滅菌 BMHB 15 mL のアルミ キャップ付き。37 ° C で 24 時間この懸濁液を育成します。</li>
			<li>標準化された接種の準備のための 4 ° C で冷蔵庫内を格納します。</li>
		</ol>
</li>
		<li>原液から (手順 2.1.1 参照)大腸菌取るの約 0.1 - 0.2 mL の細菌懸濁液とプラスチック製の 15 mL の希釈滅菌チューブ 10 mL の滅菌バッファー ミュラー ヒントン スープ (BMHB) が記入されました。24 h の 37 ° C でチューブを孵化させなさい。</li>
		<li>標準化された接種 (1.5 × 108 CFU/mL) の準備のため事前の滅菌 BMHB 10 mL でいっぱい 15 mL 滅菌チューブにこの懸濁液の少量を追加します。</li>
		<li>徹底的に渦混合物とメジャー 600 の光学濃度 (OD) 濃度計32nm。</li>
		<li>2.1.3 - 2.1.4 マクファーランド値はきれいな BMHB の値と比較して 0.5 ずつ増加を示した OD までの手順を繰り返します。</li>
	</ol>
</li>
	<li>接種の手順 2 つ異なるアルミ箔 (20 cm × 30 cm) コントロールのサンプルと接種肉サンプルの 2 番目の長方形の肉サンプルを配置します。<ol>
<li>2 番目のアルミ箔を表面に接種を均等に分散することにより (これは 1.2 × 108 CFU 肉サンプルごとに相当) 選択したひずみの細菌懸濁液の 800 μ L で生肉長方形サンプルを接種します。<ol>
<li>サンプルの 1 つの側面に 400 μ L をピペットし、無菌細胞拡散を用いた表面を軽く広げ。サンプルの 10 分反対側の懸濁液の残りの部分と同じ手順の繰り返し乾燥させます。</li>
		</ol>
</li>
	</ol>
</li>
</ol>3. 乾燥とテオの応用
<ol><li>ドライヤーに層流の安全キャビネットから長方形の肉のサンプルを含む両方のアルミ箔を転送: 各アルミ箔でカバーし、ドライヤー内サンプルを配置。</li>
	<li>標準的な実験室のドライヤーで乾燥を行います。 注: 最初に、オーブンを 55 ° Cこの手順は、20 分間続くことができます。<ol>
<li>乾燥空気相対湿度値 30-45% から 55 ° C で 6 時間コントロールのサンプルを乾燥させます。 注: 乾燥空気相対湿度値肉からの液体の蒸発の率によって時間が異なります。</li>
	</ol>
</li>
	<li>適用すると、テオのボリュームを計算し、ドライヤー ボリューム (mL/L 空気) ごとにテオのボリュームとしてエッセンシャル オイルの濃度を表します。たとえば、53 L (ドライヤーのボリューム) でテオの 1.5 mL の用量は 0.028 mL/L 空気の濃度の結果します。エシェリヒア属大腸菌のテオのマイクを決定するするには、1.5 mL (0.028 mL/L 空気)、1 mL (0.019 mL/L 空気) と 0.75 mL (0.014 mL/L 空気) の用量を使用します。</li>
	<li>テオの適用のため、乾燥する前に、(主な複合 79% としてチモール)、蒸気は、ファンの前でドライヤーにテオと場所の 1.5 mL の用量フィルター紙 (12 cm × 20 cm) を浸します。</li>
	<li>コントロールのサンプル (3.1 と 3.2 の手順) と同じ手順を使用して扱われるテオ肉サンプルを乾燥させます。 注: 乾燥のプロセスが終了し、サンプルを削除したら 80 ° C で 3 時間オーブンのスイッチ、オーブンからのエッセンシャル オイルの残留物をきれいにするため 100% 通気する空気弁表示を設定します。</li>
</ol>4. 微生物解析
<ol><li>細菌と肉接種前に任意の粗悪品のため肉のサンプルを確認します。スライムの外観は、任意の強く、刺激的な匂いの検出は、肉の腐敗を示しています。テクスチャは、ぬるぬるした感じ、細菌を始めていると、肉の表面に掛けるかもしれない。</li>
	<li>接種効率を評価するために大腸菌ATCC 25922 の存在の raw サンプルをテストし、乾燥の手順の前に非接種コントロールのサンプルとの比較をします。この目的。<ol>
<li>各肉サンプル (2 コントロールのサンプルおよび 2 サンプル) を洗います。1:10 (w/v) 7 7.3 から pH の範囲との比率でペプトン水 (8.5 g, 塩化ナトリウムのペプトン 1 g、リン酸緩衝生理食塩水、5 錠、1 L の水にポリソルベート 80 の 1 g) で滅菌フラスコ内の各肉サンプルを中断します。室温で 10 分間 140 rpm でシェーカーを使用して横に振る。 注: 接種手続き後すぐに洗ってください。</li>
		<li>プレート カウント寒天培地 (PCA) とマッコンキー寒天培地 (MCA) アーウィン ・ ネース, 陳33によって要約される調整 6 × 6 ドロップ プレート プロシージャによって細菌数を評価します。 注: 6 × 6 ドロップ方法は、マルチ チャンネル ピペット、集中的より経済的な従来法33,と比較してより少ない労働であると調査のサンプルの 10 倍のシリアル希薄を準備するスープ マイクロ希釈法を使います34。</li>
		<li>エシェリヒア属大腸菌の評価による 6 × 6 ドロップ プレート シリアル サンプルの 10 倍希釈液を栽培してください。 注: 特に 6 × 6 ドロップ方法、栽培使用 6 5 μ L-滴、6 から選択マルチ チャンネル ピペットの調査のサンプルの希釈液。適切に乾燥ペトリ皿に寒天に滴をすぐに吸収してこのメソッドによって植わることは非常に便利で管理しやすい34。</li>
		<li>24 h の 37 ° C でシャーレを孵化させなさい。栽培期間後に、セクション 5 で説明したようペトリ皿 (乾燥肉 CFU g-1 )エシェリヒア属大腸菌の細菌コロニー数を評価します。</li>
	</ol>
</li>
	<li>乾燥後 2 つ接種乾燥サンプルと実行可能な大腸菌、それぞれそれら乾燥非接種コントロールの 2 つのサンプルと比較します。これらの 4 つのサンプルのエシェリヒア属大腸菌の有無を決定するには、各肉のサンプルの前濃縮プロセスをとおり実行します。<ol>
<li>ペプトン水で滅菌フラスコ内の各肉サンプルを中断 (を参照してくださいステップ 4.2.1)、室温で 10 分間 140 rpm でシェーカーを使用して横に振る。その後、前濃縮 6 h 中に 37 ° C で各フラスコを孵化させなさい。</li>
		<li>細菌の培養と評価の手順 4.2.2 - 4.2.4 で同じ手順に従います。</li>
	</ol>
</li>
</ol>5. 結果を確認します。
<ol><li>孵化が完了したら、インキュベーターからペトリ皿を削除し、次のように結果を確認します。<ol>
<li>PCA (白斑) と典型的なエシェリヒア属大腸菌の中温好気性細菌の存在のプレートを調べるコロニー数の合計を評価する MCA の植民地 (濃いピンクに赤)。病原体が存在しない場合、両方の寒天は成長はありません。</li>
		<li>コロニーをカウントし、大腸菌(CFU/g-1乾燥肉の) 現在の量を決定します。 注: は、ドロップ (図 1) 当たり 30 以下の植民地を含む 2 つの連続希薄でコロニー (N) の数をカウントします。次のとおり35として乾燥肉の CFU/g-1の数Nを決定します。 <img alt="Equation" src="/files/ftp_upload/57054/57054eq1.jpg"> どこで、 Cはカウントすべての滴のコロニーの合計、 vは、サンプル希釈使用ドロップ (ここでは、0.05 mL) あたりの体積、 n1は最初の希釈で使用する滴数、 n2は、液滴の数2 番目の希釈で使用、 dは、最初のカウントがキャプチャされた希釈を表します。</li>
	</ol>
</li>
	<li>微生物学的データを解析するには、ログ CFU g-1と治療36の主な効果の分散分析 (ANOVA) にそれらを服従にコロニー数を変換します。<ol>
<li>複数の平均比較36テューキー正直差テスト (テューキー HSD) を行い、治療の重要な違いを特定します。</li>
	</ol>
</li>
</ol>

<ol>
	<li>Eklund, M. W., Peterson, M. E., Poysky, F. T., Paranjpye, R. N., Pelroy, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Control+of+bacterial+pathogens+during+processing+of+cold-smoked+and+dried+salmon+strips.">Control of bacterial pathogens during processing of cold-smoked and dried salmon strips.</a> J. Food Prot. 67 (2), 347-351 (2004).</li><li>Mahmoud, B. S. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preservative+effect+of+combined+treatment+with+electrolyzed+NaCl+solutions+and+essential+oil+compounds+on+carp+fillets+during+convectional+air-drying.">Preservative effect of combined treatment with electrolyzed NaCl solutions and essential oil compounds on carp fillets during convectional air-drying.</a> Int. J. Food Microbiol. 106 (3), 331-337 (2006).</li><li>Rahman, M. S., Guizani, N., Al-Ruzeiki, M. H., Al Khalasi, A. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microflora+Changes+in+Tuna+Mince+During+Convection+Air+Drying.">Microflora Changes in Tuna Mince During Convection Air Drying.</a> Dry. Technol. 18 (10), 2369-2379 (2000).</li><li>Faith, N. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Viability+of+Escherichia+coli+O157:+H7+in+ground+and+formed+beef+jerky+prepared+at+levels+of+5+and+20%+fat+and+dried+at+52,+57,+63,+or+68+C+in+a+home-style+dehydrator.">Viability of Escherichia coli O157: H7 in ground and formed beef jerky prepared at levels of 5 and 20% fat and dried at 52, 57, 63, or 68 C in a home-style dehydrator.</a> Int. J. Food Microbiol. 41 (3), 213-221 (1998).</li><li>Hierro, E., De La Hoz, L., Ord&#243;&#241;ez, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Headspace+volatile+compounds+from+salted+and+occasionally+smoked+dried+meats+(cecinas)+as+affected+by+animal+species.">Headspace volatile compounds from salted and occasionally smoked dried meats (cecinas) as affected by animal species.</a> Food Chem. 85 (4), 649-657 (2004).</li><li>Nummer, B. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+Preparation+Methods+on+the+Microbiological+Safety+of+Home-Dried+Meat+Jerky.">Effects of Preparation Methods on the Microbiological Safety of Home-Dried Meat Jerky.</a> J. Food Prot. 67 (10), 2337-2341 (2004).</li><li>Greig, J. D., Ravel, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+foodborne+outbreak+data+reported+internationally+for+source+attribution.">Analysis of foodborne outbreak data reported internationally for source attribution.</a> Int. J. Food Microbiol. 130 (2), 77-87 (2009).</li><li>Eidson, M., Sewell, C. M., Graves, G., Olson, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beef+jerky+gastroenteritis+outbreaks.">Beef jerky gastroenteritis outbreaks.</a> J. Environ. Health. 62 (6), 9-13 (2000).</li><li>Allen, K., Cornforth, D., Whittier, D., Vasavada, M., Nummer, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+high+humidity+and+wet+marinade+methods+for+pasteurization+of+jerky.">Evaluation of high humidity and wet marinade methods for pasteurization of jerky.</a> J. Food Sci. 72 (7), (2007).</li><li>Levine, P., Rose, B., Green, S., Ransom, G., Hill, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathogen+testing+of+ready-to-eat+meat+and+poultry+products+collected+at+federally+inspected+establishments+in+the+United+States,+1990+to+1999.">Pathogen testing of ready-to-eat meat and poultry products collected at federally inspected establishments in the United States, 1990 to 1999.</a> J. Food Prot. 64 (8), 1188-1193 (1990).</li><li>Keene, W. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+outbreak+of+Escherichia+coli+O157:H7+infections+traced+to+jerky+made+from+deer+meat.">An outbreak of Escherichia coli O157:H7 infections traced to jerky made from deer meat.</a> JAMA. 277 (15), 1229-1231 (1997).</li><li>Oliver, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+viable+but+nonculturable+state+in+bacteria.">The viable but nonculturable state in bacteria.</a> J. Microbiol. 43, 93-100 (2005).</li><li>Oliver, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+findings+on+the+viable+but+nonculturable+state+in+pathogenic+bacteria.">Recent findings on the viable but nonculturable state in pathogenic bacteria.</a> FEMS Microbiol. Rev. 34 (4), 415-425 (2010).</li><li>Khamisse, E., Firmesse, O., Christieans, S., Chassaing, D., Carpentier, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+cleaning+and+disinfection+on+the+non-culturable+and+culturable+bacterial+loads+of+food-contact+surfaces+at+a+beef+processing+plant.">Impact of cleaning and disinfection on the non-culturable and culturable bacterial loads of food-contact surfaces at a beef processing plant.</a> Int. J. Food Microbiol. 158 (2), 163-168 (2012).</li><li>Li, L., Mendis, N., Trigui, H., Oliver, J. D., Faucher, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+importance+of+the+viable+but+non-culturable+state+in+human+bacterial+pathogens.">The importance of the viable but non-culturable state in human bacterial pathogens.</a> Front. Microbiol. 5, 258 (2014).</li><li>Hern&#225;ndez, H., Claramount, D., Ku&#269;erov&#225;, I., Banout, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+modified+blanching+and+oregano+essential+oil+on+drying+kinetics+and+sensory+attributes+of+dried+meat.">The effects of modified blanching and oregano essential oil on drying kinetics and sensory attributes of dried meat.</a> J. Food Process. Preserv. , (2016).</li><li>Garc&#237;a-D&#237;ez, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Impact+of+Essential+Oils+on+Consumer+Acceptance+of+Chouri&#231;o+de+vinho+-+A+Dry-Cured+Sausage+Made+from+Wine-Marinated+Meat+-+Assessed+by+the+Hedonic+Scale,+JAR+Intensity+Scale+and+Consumers'+&amp;#34;Will+to+Consume+and+Purchase.&amp;#34.">The Impact of Essential Oils on Consumer Acceptance of Chouri&#231;o de vinho - A Dry-Cured Sausage Made from Wine-Marinated Meat - Assessed by the Hedonic Scale, JAR Intensity Scale and Consumers' &amp;#34;Will to Consume and Purchase.&amp;#34.</a> J. Food Process. Preserv. 41 (4), (2017).</li><li>Govaris, A., Solomakos, N., Pexara, A., Chatzopoulou, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+antimicrobial+effect+of+oregano+essential+oil,+nisin+and+their+combination+against+Salmonella+Enteritidis+in+minced+sheep+meat+during+refrigerated+storage.">The antimicrobial effect of oregano essential oil, nisin and their combination against Salmonella Enteritidis in minced sheep meat during refrigerated storage.</a> Int. J. Food Microbiol. 137 (2-3), 175-180 (2010).</li><li>Holley, R. A., Patel, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improvement+in+shelf-life+and+safety+of+perishable+foods+by+plant+essential+oils+and+smoke+antimicrobials.">Improvement in shelf-life and safety of perishable foods by plant essential oils and smoke antimicrobials.</a> Food Microbiol. 22 (4), 273-292 (2005).</li><li>Petrou, S., Tsiraki, M., Giatrakou, V., Savvaidis, I. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chitosan+dipping+or+oregano+oil+treatments,+singly+or+combined+on+modified+atmosphere+packaged+chicken+breast+meat.">Chitosan dipping or oregano oil treatments, singly or combined on modified atmosphere packaged chicken breast meat.</a> Int. J. Food Microbiol. 156 (3), 264-271 (2012).</li><li>Ballester-costa, C., Sendra, E., Viuda-martos, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+Antioxidant+and+Antibacterial+Properties+on+Meat+Homogenates+of+Essential+Oils+Obtained+from+Four+Thymus+Species+Achieved+from+Organic+Growth.">Assessment of Antioxidant and Antibacterial Properties on Meat Homogenates of Essential Oils Obtained from Four Thymus Species Achieved from Organic Growth.</a> Foods. 6 (8), 59 (2017).</li><li>Hern&#225;ndez, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+oregano+essential+oil+on+microbial+load+and+sensory+attributes+of+dried+meat.">The effect of oregano essential oil on microbial load and sensory attributes of dried meat.</a> J. Sci. Food Agric. 97 (1), 82-87 (2017).</li><li>Garc&#237;a-D&#237;ez, J., Alheiro, J., Falco, V., Fraqueza, M. J., Patarata, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+characterization+and+antimicrobial+properties+of+herbs+and+spices+essential+oils+against+pathogens+and+spoilage+bacteria+associated+to+dry-cured+meat+products.">Chemical characterization and antimicrobial properties of herbs and spices essential oils against pathogens and spoilage bacteria associated to dry-cured meat products.</a> J. Essent. Oil Res. 29 (2), 117-125 (2017).</li><li>Cavanagh, H. M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antifungal+Activity+of+the+Volatile+Phase+of+Essential+Oils:+A+Brief+Review.">Antifungal Activity of the Volatile Phase of Essential Oils: A Brief Review.</a> Nat. Prod. Commun. 2 (12), 1297-1302 (2007).</li><li>Tajkarimi, M. M., Ibrahim, S. A., Cliver, D. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antimicrobial+herb+and+spice+compounds+in+food.">Antimicrobial herb and spice compounds in food.</a> Food Control. 21 (9), 1199-1218 (2010).</li><li>Nedorostova, L., Kloucek, P., Kokoska, L., Stolcova, M., Pulkrabek, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antimicrobial+properties+of+selected+essential+oils+in+vapour+phase+against+foodborne+bacteria.">Antimicrobial properties of selected essential oils in vapour phase against foodborne bacteria.</a> Food Control. 20 (2), 157-160 (2009).</li><li>Burt, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Essential+oils:+Their+antibacterial+properties+and+potential+applications+in+foods+-+A+review.">Essential oils: Their antibacterial properties and potential applications in foods - A review.</a> Int. J. Food Microbiol. 94 (3), 223-253 (2004).</li><li>Ramanathan, L., Das, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studies+on+the+control+of+lipid+oxidation+in+ground+fish+by+some+polyphenolic+natural+products.">Studies on the control of lipid oxidation in ground fish by some polyphenolic natural products.</a> J. Agric. Food Chem. 40 (1), 17-21 (1992).</li><li>Yamazaki, K., Yamamoto, T., Kawai, Y., Inoue, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhancement+of+antilisterial+activity+of+essential+oil+constituents+by+nisin+and+diglycerol+fatty+acid+ester.">Enhancement of antilisterial activity of essential oil constituents by nisin and diglycerol fatty acid ester.</a> Food Microbiol. 21 (3), 283-289 (2004).</li><li>Garc&#237;a-D&#237;ez, J., Alheiro, J., Falco, V., Fraqueza, M. J., Patarata, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synergistic+activity+of+essential+oils+from+herbs+and+spices+used+on+meat+products+against+food+borne+pathogens.">Synergistic activity of essential oils from herbs and spices used on meat products against food borne pathogens.</a> Nat. Prod. Commun. 12 (2), 281-286 (2017).</li><li>Hussein Hamdy Roby, M., Atef Sarhan, M., Abdel-Hamed Selim, K., Ibrahim Khalel, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+antioxidant+activity,+total+phenols+and+phenolic+compounds+in+thyme+(Thymus+vulgaris+L.),+sage+(Salvia+officinalis+L.),+and+marjoram+(Origanum+majorana+L.)+extracts.">Evaluation of antioxidant activity, total phenols and phenolic compounds in thyme (Thymus vulgaris L.), sage (Salvia officinalis L.), and marjoram (Origanum majorana L.) extracts.</a> Ind. Crops Prod. 43, 827-831 (2013).</li><li>Gouveia, A. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Antimicrobial+Effect+of+Essential+Oils+Against+Listeria+monocytogenes+in+Sous+vide+Cook-Chill+Beef+during+Storage.">The Antimicrobial Effect of Essential Oils Against Listeria monocytogenes in Sous vide Cook-Chill Beef during Storage.</a> J. Food Process. Preserv. 41 (4), (2017).</li><li>Chen, C., Nace, G., Irwin, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+6+x+6+drop+plate+method+for+simultaneous+colony+counting+and+MPN+enumeration+of+Campylobacter+jejuni,+Listeria+monocytogenes,+and+Escherichia+coli.">A 6 x 6 drop plate method for simultaneous colony counting and MPN enumeration of Campylobacter jejuni, Listeria monocytogenes, and Escherichia coli.</a> J. Microbiol. Methods. 55 (2), 475-479 (2003).</li><li>Herigstad, B., Hamilton, M., Heersink, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+to+optimize+the+drop+plate+method+for+enumerating+bacteria.">How to optimize the drop plate method for enumerating bacteria.</a> J. Microbiol. Methods. 44 (2), 121-129 (2001).</li><li>. Practical food microbiology Available from: <a target="_blank" href="https://drive.google.com/file/d/0BzyVOLllJ0B1YmlEemZ5M1RZekU/view?ts=590d8019">https://drive.google.com/file/d/0BzyVOLllJ0B1YmlEemZ5M1RZekU/view?ts=590d8019</a> (2003)</li><li>Smith-Palmer, A., Stewart, J., Fyfe, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antimicrobial+properties+of+plant+essential+oils+and+essences+against+five+important+food-borne+pathogens.">Antimicrobial properties of plant essential oils and essences against five important food-borne pathogens.</a> Lett. Appl. Microbiol. 26 (2), 118-122 (1998).</li><li>Burt, S. a., Reinders, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antibacterial+activity+of+selected+plant+essential+oils+against+Escherichia+coli+O157:H7.">Antibacterial activity of selected plant essential oils against Escherichia coli O157:H7.</a> Lett. Appl. Microbiol. 36 (3), 162-167 (2003).</li></ol>

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以前の研究では、乾燥10年食中毒の原因微生物間生き残ることを示しています。したがって、食品の安全性を確保するために乾燥する前に防腐剤を適用する必要は。本研究で我々 はテオを使用してに焦点を当てます。理由は 2 つあります: 最初に、代替添加剤として天然物を使用して食品品質16; を向上させる消費者からの高い需要があります。第二に、以?...

乾燥食品を保存する伝統的な方法としては、古くから使用されています。最近では、乾燥食品保全1,2,3の効果的な方法としての関心の高まりがあります。それは様々 な特殊加工肉を作るためです。最もよく知られているの 1 つはジャーキーです。
ジャーキー、肉保存のための最も古い方法の 1 つは、硬化・乾燥を行う低水分活性に基づいて、したがってその寿命4を拡張します。今日では、保存塩漬け肉は非常に人気があるまだぎくしゃくした、食品の安全性、味、テクスチャが必須です。肉は、牛肉、豚肉、鶏肉、またはゲーム5を含むほぼすべてのタイプのけいれん的な準備が使え、無駄のないストリップで肉をまな板と乾燥が必要です。通常、硬化ソリューションまたは喫煙で肉をマリネ乾燥と共に得られるぎくしゃくの独特の味6。
本当に食べ物を保存する乾燥の広大な関心にもかかわらず不十分な乾燥肉から大腸菌による食中毒発生の危険性は非常に重要で、制御する必要があります。いくつかの研究は、特にE. 大腸菌o157: h7、ホーム乾燥中の処理不十分な熱に帰因すると食中毒胃腸炎アウトブレイクの報告があります。似たようなケースは、ぎくしゃくした7,8,9を商業的準備でも発生しています。レバイン氏ら。10は、食中毒微生物が商業のぎくしゃくした生産者によって使用される適度な乾燥条件 (約 60 ° C) を生き残ることができることを提案しました。エシェリヒア属大腸菌o157: h7、1990 年代半ばに食中毒の発生は、乾燥させてすりつぶした肉製品6,11に帰因しました。興味深いことに、すべての前の例で主なリスクが認められた非人為的 (VBNC) が実行可能な細菌の病原体が原因です。大腸菌の細胞は飢餓や温度変化など様々 なストレスの下で、VBNC 状態12,13として知られている特定の状態を入力しました。VBNC セル適切な条件への露出によって培養細胞に戻る、蘇生可能性があり、食中毒汚染14,15による人の健康への脅威。これは、製品を乾燥させた直後に肉を消費した場合は、安全を意味します。ただし、湿度が高くなどの不適切なストレージの場合、病原体や微生物の増殖の再活性化のリスクが高いがあります。
乾燥とマリネの方法に加えて、食品品質16,17を改善する添加物の代替として自然の製品を使用する消費者からの高い需要があります。古典的な合成防腐剤18,19,20,21の代わりに肉の天然食品添加物のアプリケーションに特に興味があった。肉を乾燥エッセンシャル オイルの使用に十分な実験的証拠の欠乏がある、にもかかわらず、この分野の初期の研究は既に肯定的な結果22,23を示します。
中間年齢以来、人々 は、抗菌、殺虫、駆虫特性24,25,26精油化合物 (Eoc) を認識しています。今日、Eoc 生理活性天然化合物の最も重要なグループの 1 つの一部であります。異なる Eoc のチモールは、最もよく知られているの 1 つです。それはテオ23の 85% 以上で構成されます。このフェノールは、食品に追加される微生物・化学的劣化を防ぎます。さらに、その他天然の防腐剤2,27,の28,29,30との組み合わせでその抗菌特性を改善可能性があります。タイム (タチジャコウソウ)、今日では、シソ科の家族に属しているハーブ香料剤として非常に効果的な肉防腐剤31として認識されています。ガルシーア ・ ディーツらによる研究。30肉製品には、テオに他のエッセンシャル オイルと比較した場合の食品由来病原体に対する広い阻害パターンが表示されることを発見しました。したがって、乾燥肉の価値を増加し、乾燥処理中にエッセンシャル オイルを適用することによって媒介する病気のリスクを軽減する機会があります。
このプロトコルでは乾燥肉中にテオを適用する手法を提案する、商工会議所、乾燥蒸気フォームで直接で具体的に。評価、原料のものと比較して扱われたサンプルで病原性の細菌の有無を決定するためにマイクを使用します。予備の結果は、このメソッドが合成防腐剤に非常に効果的な代替手段であり、この乾燥肉の微生物負荷が大幅に減少することを示します。

<table><tbody><tr><td>Meat cutter</td><td>Kalorik</td><td>KP 3530</td><td>from Miami Gardens, FL, USA</td></tr><tr><td>Laminar safety cabinet</td><td>Faster s.r.l</td><td></td><td>from Italy</td></tr><tr><td>Squeeze bottle of 500 mL</td><td>Merci</td><td>632 524 325 025</td><td>from CZ</td></tr><tr><td>Standard laboratory drier UFE 400</td><td>Memmert</td><td>DE 66812464</td><td>from Germany</td></tr><tr><td>Incubator</td><td>BT 120</td><td>N/A</td><td>from CZ</td></tr><tr><td>Refrigerator and Freezer</td><td>Bosch</td><td>KGN34VW20G</td><td>from DE</td></tr><tr><td>Densitometer</td><td>Biosan</td><td>220 000 050 122</td><td>Latvia; supplier Merci, CZ</td></tr><tr><td>&lt;em&gt;Escherichia coli&lt;/em&gt; ATCC 25922</td><td>Oxoid</td><td>CL7050</td><td>from CZ</td></tr><tr><td>Vortex</td><td>Chromservis</td><td>22008013</td><td>from CZ</td></tr><tr><td>Sterilized plastic tubes 15 mL</td><td>Gama</td><td>331 000 020 115</td><td>from CZ, supplier Merci</td></tr><tr><td>20 mL injection vial</td><td>Healthy vial</td><td>hvft169</td><td>from China</td></tr><tr><td>20 mm sterile butyl rubber stopper</td><td>Merci</td><td>22008013</td><td>from CZ</td></tr><tr><td>20 mm aluminum cap</td><td>Healthy vial</td><td>N/A</td><td>from China</td></tr><tr><td>Thyme essential oil</td><td>Sigma Aldrich</td><td>W306509</td><td>from St Louis, MO, USA</td></tr><tr><td>Mueller Hinton Broth</td><td>Oxoid</td><td>CM0337</td><td>from CZ</td></tr><tr><td>NaCl</td><td>Penta</td><td>16610-31000</td><td>from CZ</td></tr><tr><td>Peptone</td><td>Oxoid</td><td>LP0034</td><td>from CZ</td></tr><tr><td>Phosphate-buffered saline</td><td>Sigma Aldrich</td><td>P4417</td><td>from CZ</td></tr><tr><td>Polysorbate 80 (Tween 80)</td><td>Roth</td><td>T 13502</td><td>from DE, supplier P-lab</td></tr><tr><td>Shaker SHO-1D</td><td>Verkon</td><td>DH.WSR04020</td><td>from CZ,&amp;nbsp; 10 - 300 rpm. 350 x 350 mm with a platform for flasks</td></tr><tr><td>Ethanol 70%</td><td>Bioferm</td><td>N/A</td><td>from CZ</td></tr><tr><td>MacConkey Agar</td><td>Oxoid</td><td>CM007</td><td>from CZ</td></tr><tr><td>Plate Count Agar</td><td>Oxoid</td><td>CM0325</td><td>from CZ</td></tr><tr><td>Filter paper</td><td>Merci</td><td>480 622 080 040</td><td>from CZ</td></tr><tr><td>Erlenmeyer flasks 250 mL</td><td>Simax</td><td>610 002 122 636</td><td>from CZ; supplier Merci CZ</td></tr><tr><td>Multichannel pipette</td><td>Socorex</td><td>S852820</td><td>from Switzerland; supplier P lab, CZ</td></tr><tr><td>Microtiter plate</td><td>Gamma</td><td>V400916</td><td>CZ</td></tr><tr><td>Microlitre pipette 100-1000 &amp;mu;L</td><td>Eppendorf</td><td>333 120 000 062</td><td>from Germany; supplier Merci, CZ</td></tr></tbody></table>

the effect of the application of thyme essential oil on microbial load during meat drying

肉は、保全と安全性が重要なぎくしゃくした、人気のある食品おやつの準備で使用されている、高たんぱくな食事です。食品の安全性を確保するために、肉及び肉製品の棚寿命を延ばすため、合成又は天然の防腐剤の使用はコントロールに適用されているし、食中毒細菌を除去します。肉の天然食品添加物のアプリケーションの関心の高まりが増加しています。肉及び肉製品、食中毒を引き起こす大腸菌などの微生物を汚染します。したがって、肉保全プロセスを改善する必要があります。しかし、肉を乾燥するときのエッセンシャル オイルの使用は深く研究されていません。この点で、乾燥肉の価値を増加し、乾燥処理中にエッセンシャル オイルを適用することによって媒介する病気のリスクを軽減する機会があります。このプロトコルでは、乾燥、乾燥室で直接蒸気の形で具体的には肉の中にタイムの精油 (テオ) を適用する手法を提案する.評価は、最小阻止濃度 (MIC) を使用して、raw のサンプルと比較して扱われたサンプルで有害な細菌の数を検出します。予備的な結果は、このメソッドは、実行可能な代替オプション合成防腐剤を示し乾燥肉の微生物負荷が大幅に減少すること。

まず以前食品の安全性を高めるため、乾燥肉の価値を増加するオレガノ精油 (OEO) を使用してこのメソッドを開発した.一般に、前の実験では大腸菌が生存戦略として乾燥時 VBNC 状態に入る。22乾燥後、培養菌はなかったという事実によってこれを示します。したがって、6 h の前濃縮プロセスひずみのカウントを許可する必要があります。短い?...

Watch this Scientific Journal Video about The Effect of the Application of Thyme Essential Oil on Microbial Load During Meat Drying at JoVE.com

The Effect of the Application of Thyme Essential Oil on Microbial Load During Meat Drying

タイムのエッセンシャル オイルのアプリケーションの肉乾燥中の微生物負荷に及ぼす影響

肉製品を汚染する大腸菌などの微生物は、食中毒を引き起こします。肉の乾燥工程でのエッセンシャル オイルの使用は深く研究されていません。ここでは、乾燥肉の微生物負荷を低減する乾燥時に肉にタイムのエッセンシャル オイルを適用する手法を提案する.

Meat is a high protein meal that is used in the preparation of jerky, a popular food snack, where preservation and safety are important. To assure food ...

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9.2K ビュー.  Czech University of Life Sciences Prague. 肉製品を汚染する大腸菌などの微生物は、食中毒を引き起こします。肉の乾燥工程でのエッセンシャル オイルの使用は深く研究されていません。ここでは、乾燥肉の微生物負荷を低減する乾燥時に肉にタイムのエッセンシャル オイルを適用する手法を提案する.

ビデオ: The Effect of the Application of Thyme Essential Oil on Microbial Load During Meat Drying

Preparation of Homogeneous MALDI Samples for Quantitative Applications

This protocol demonstrates a simple sample preparation to reduce spatial heterogeneity in ion signals during matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The heterogeneity of ion signals is a severe problem in MALDI, which results in poor data reproducibility and makes MALDI unsuitable for quantitative analysis. By regulating sample plate temperature during sample preparation, thermal-induced hydrodynamic flows inside droplets of sample solution are able to reduce the heterogeneity problem. A room-temperature sample preparation chamber equipped with a temperature-regulated copper base block that holds MALDI sample plates facilitates precise control of the sample drying condition. After drying of sample droplets, the temperature of sample plates is returned to room temperature and removed from the chamber for subsequent mass spectrometric analysis. The areas of samples are examined with MALDI-imaging mass spectrometry to obtain the spatial distribution of all components in the sample. In comparison with the conventional dried-droplet method that prepares samples under ambient conditions without temperature control, the samples prepared with the method demonstrated herein show significantly better spatial distribution and signal intensity. According to observations using carbohydrate and peptide samples, decreasing substrate temperature while maintaining the surroundings at ambient temperature during the drying process can effectively reduce the heterogeneity of ion signals. This method is generally applicable to various combinations of samples and matrices.

A protocol for reducing spatial heterogeneities of ion signals in MALDI mass spectrometry by regulating substrate temperature during sample drying processes is demonstrated.

定量的なアプリケーションのための均質なMALDI試料の調製

This protocol demonstrates a simple sample preparation to reduce spatial heterogeneity in ion signals during matrix-assisted laser desorption/ionization ...

生化学

Sample Preparation of Mycobacterium tuberculosis Extracts for Nuclear Magnetic Resonance Metabolomic Studies

Mycobacterium tuberculosis is a major cause of mortality in human beings on a global scale. The emergence of both multi- (MDR) and extensively-(XDR) drug-resistant strains threatens to derail current disease control efforts. Thus, there is an urgent need to develop drugs and vaccines that are more effective than those currently available. The genome of M. tuberculosis has been known for more than 10 years, yet there are important gaps in our knowledge of gene function and essentiality. Many studies have since used gene expression analysis at both the transcriptomic and proteomic levels to determine the effects of drugs, oxidants, and growth conditions on the global patterns of gene expression. Ultimately, the final response of these changes is reflected in the metabolic composition of the bacterium including a few thousand small molecular weight chemicals. Comparing the metabolic profiles of wild type and mutant strains, either untreated or treated with a particular drug, can effectively allow target identification and may lead to the development of novel inhibitors with anti-tubercular activity. Likewise, the effects of two or more conditions on the metabolome can also be assessed. Nuclear magnetic resonance (NMR) is a powerful technology that is used to identify and quantify metabolic intermediates. In this protocol, procedures for the preparation of M. tuberculosis cell extracts for NMR metabolomic analysis are described. Cell cultures are grown under appropriate conditions and required Biosafety Level 3 containment,1 harvested, and subjected to mechanical lysis while maintaining cold temperatures to maximize preservation of metabolites. Cell lysates are recovered, filtered sterilized, and stored at ultra-low temperatures. Aliquots from these cell extracts are plated on Middlebrook 7H9 agar for colony-forming units to verify absence of viable cells. Upon two months of incubation at 37 &deg;C, if no viable colonies are observed, samples are removed from the containment facility for downstream processing. Extracts are lyophilized, resuspended in deuterated buffer and injected in the NMR instrument, capturing spectroscopic data that is then subjected to statistical analysis. The procedures described can be applied for both one-dimensional (1D) 1H NMR and two-dimensional (2D) 1H-13C NMR analyses. This methodology provides more reliable small molecular weight metabolite identification and more reliable and sensitive quantitative analyses of cell extract metabolic compositions than chromatographic methods. Variations of the procedure described following the cell lysis step can also be adapted for parallel proteomic analysis.

Sample Preparation of Mycobacterium tuberculosis Extracts for Nuclear Magnetic Resonance Metabolomic Studies

The metabolomic profile of Mycobacterium tuberculosis is determined after growth in broth cultures. Conditions can be varied to test the effects of nutritional supplements, oxidants, and anti-tuberculosis agents on the metabolic profile of this microorganism. Procedure for extract preparation is applicable for both 1D 1H and 2D 1H-13C NMR analyses.

のサンプル調製<em&gt;結核菌</em核磁気共鳴代謝研究のための&gt;抽出

Mycobacterium tuberculosis is a major cause of mortality in human beings on a global scale. The emergence of both multi- (MDR) and extensively-(XDR) ...

免疫・感染学

Measuring Volatile and Non-volatile Antifungal Activity of Biocontrol Products

The protocol described is based on a plug-transfer technique that allows accurate determination of microorganism quantities and their developmental stages. A specified number of spores are spread on an agar plate. This agar plate is incubated for a defined period to allow the fungi to reach the expected developmental stage, except for spores where incubation is not required. Agar plugs covered by spores, hyphae, or mycelium are next withdrawn and transferred onto agar media containing the antifungal compound to be tested either placed at a distance from the fungi or in contact. This method is applicable to test both liquid extracts and solid samples (powders). It is particularly well suited for quantifying the relative contributions of volatile and non-volatile agents in bioactive mixtures and for determining their effects, specifically on spores, early hyphae, and mycelium.
The method is highly relevant for the characterization of the antifungal activity of biocontrol products, notably plant-derived products. Indeed, for plant treatment, the results can be used to guide the choice of mode of application and to establish the trigger thresholds.

We describe a modified agar-based method designed to quantify the antifungal effects of plant-derived products. Both volatile and non-volatile contributions to the antifungal activity can be assessed through this protocol. In addition, efficacy against fungi can be measured at key developmental stages in a single experimental setup.

バイオコントロール製品の揮発性および不揮発性抗真菌活性の測定

The protocol described is based on a plug-transfer technique that allows accurate determination of microorganism quantities and their developmental ...

環境学

Investigation of Microbial Cooperation via Imaging Mass Spectrometry Analysis of Bacterial Colonies Grown on Agar and in Tissue During Infection

Understanding the metabolic consequences of microbial interactions that occur during infection presents a unique challenge to the field of biomedical imaging. Matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry represents a label-free, in situ imaging modality capable of generating spatial maps for a wide variety of metabolites. While thinly sectioned tissue samples are now routinely analyzed via this technology, imaging mass spectrometry analyses of non-traditional substrates, such as bacterial colonies commonly grown on agar in microbiology research, remain challenging due to the high water content and uneven topography of these samples. This paper demonstrates a sample preparation workflow to allow for imaging mass spectrometry analyses of these sample types. This process is exemplified using bacterial co-culture macrocolonies of two gastrointestinal pathogens: Clostridioides difficile and Enterococcus faecalis. Studying microbial interactions in this well-defined agar environment is also shown to complement tissue studies aimed at understanding microbial metabolic cooperation between these two pathogenic organisms in mouse models of infection. Imaging mass spectrometry analyses of the amino acid metabolites arginine and ornithine are presented as representative data. This method is broadly applicable to other analytes, microbial pathogens or diseases, and tissue types where a spatial measure of cellular or tissue biochemistry is desired.

Investigation of Microbial Cooperation via Imaging Mass Spectrometry Analysis of Bacterial Colonies Grown on Agar and in Tissue During Infection

A novel sample preparation method is demonstrated for the analysis of agar-based, bacterial macrocolonies via matrix-assisted laser desorption/ionization imaging mass spectrometry.

イメージング質量分析 による 寒天上および感染組織内で増殖した細菌コロニーの微生物連携の検討

Understanding the metabolic consequences of microbial interactions that occur during infection presents a unique challenge to the field of biomedical ...

Therapeutic Effectiveness of a Dietary Supplement for Management of Halitosis in Dogs

Halitosis is a common complaint involving social and communicational problems in humans and also affects the pet-owner relationship. In this randomized placebo-controlled crossover clinical evaluation, we assessed&#160;the effectiveness of a dedicated dietary supplement to improve chronic halitosis in 32 dogs of different breeds and ages. This protocol describes how to evalute the presence of oral volatile suphur compunds, e.g. methyl mercaptan, hydrogen sulfide and dimethyl sulfide, by means of a portable gas chromatograph device coupled with a syringe, which was used to collect the breath, and a dedicated software, which allows the operator to monitor each compound concentration during each measurement, in a relatively short time (8 min).
A significant modification of halitosis parameters was observed after 30 days since the beginning of treatment (p &#60;0.05), while a long-lasting effect was still observed even 20 days after the suspension of the treatment. Portable gas chromatograph, which is also widely used in clinical practice, can be therefore used to confirm&#160;and control halitosis in humans and animals. Even though human and animal species present some differences,&#160;this innovative and alternative therapy for halitosis management might be extended to human clinical practice as an adjuvant dietary approach.

We describe a simple approach for diagnosis of halitosis in dogs as well as a dietary approach for its management. This protocol may be extended to the management of halitosis in humans in the near future.

犬の口臭の管理のための栄養補助食品の治療有効性

Halitosis is a common complaint involving social and communicational problems in humans and also affects the pet-owner relationship. In this randomized ...

医学

Metabolic Profiling to Determine Bactericidal or Bacteriostatic Effects of New Natural Products using Isothermal Microcalorimetry

Due to the global threat of rising antimicrobial resistance, novel antibiotics are urgently needed. We investigate natural products from Myxobacteria as an&#160;innovative source of such new compounds. One bottleneck in the process is typically the elucidation of their mode-of-action. We recently established isothermal microcalorimetry as part of a routine profiling pipeline. This technology allows for investigating the effect of antibiotic exposure on the total bacterial metabolic response, including processes that are decoupled from biomass formation. Importantly, bacteriostatic and bactericidal effects are easily distinguishable without any user intervention during the measurements. However, isothermal microcalorimetry is a rather new approach and applying this method to different bacterial species usually requires pre-evaluation of suitable measurement conditions. There are some reference thermograms available of certain bacteria, greatly facilitating interpretation of results. As the pool of reference data is steadily growing, we expect the methodology to have increasing impact in the future and expect it to allow for in-depth fingerprint analyses enabling the differentiation of antibiotic classes.

The elucidation of the mode-of-action of a novel antibiotic is a challenging task in the drug discovery process. The goal of the method described here is the application of isothermal microcalorimetry using calScreener in antibacterial profiling to provide additional insight into drug-microbe interactions.

等熱マイクロカロリメトリーを用いた新天然物の殺菌効果または静電効果を測定する代謝プロファイリング

Due to the global threat of rising antimicrobial resistance, novel antibiotics are urgently needed. We investigate natural products from Myxobacteria as ...

生物工学

A Tuberculosis Molecular Bacterial Load Assay (TB-MBLA)

Tuberculosis is caused by Mycobacterium tuberculosis (Mtb), a pathogen classified by the United Nations (UN) as a dangerous category B biological substance. For the sake of the workers&#8217; safety, handling of all samples presumed to carry Mtb must be conducted in a containment level (CL) 3 laboratory. The TB molecular bacterial load assay (TB-MBLA) test is a reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) test that quantifies Mtb bacillary load using primers and dual-labelled probes for 16S rRNA. We describe the use of heat inactivation to render TB samples noninfectious while preserving RNA for the TB-MBLA. A 1 mL aliquot of the sputum sample in tightly closed 15 mL centrifuge tubes is boiled for 20 min at either 80 &#176;C, 85 &#176;C, or 95 &#176;C to inactivate Mtb bacilli. Cultivation of the heat inactivated and control (live) samples for 42 days confirmed the death of TB. The inactivated sample is then spiked with 100 &#181;L of the extraction control and RNA is extracted following the standard RNA isolation procedure. No growth was observed in the cultures of heat treated samples. The isolated RNA is subjected to real-time RT-qPCR, which amplifies a specific target in the Mtb 16S rRNA gene, yielding results in the form of quantification cycles (Cq). A standard curve is used to translate Cq into bacterial load, or estimated colony forming units per mL (eCFU/mL). There is an inverse relationship between Cq and the bacterial load of a sample. The limitation is that heat inactivation lyses some cells, exposing the RNA to RNases that cause a loss of &#60;1 log10eCFU/mL (i.e., &#60;10 CFU/mL). Further studies will determine the proportion of very low burden patients that cause false negative results due to heat inactivation.

A Tuberculosis Molecular Bacterial Load Assay TB-MBLA

We describe a tuberculosis molecular bacterial load assay test performed after heat inactivation of sputum. Heat inactivation renders sputum samples noninfectious and obviates the need for containment level 3 laboratories for tuberculosis molecular tests.

結核分子細菌負荷アッセイ(TB-MBLA)

Tuberculosis is caused by Mycobacterium tuberculosis (Mtb), a pathogen classified by the United Nations (UN) as a dangerous category B biological ...

Preparation of Prokaryotic and Eukaryotic Organisms Using Chemical Drying for Morphological Analysis in Scanning Electron Microscopy (SEM)

Scanning electron microscopy (SEM) is a widely available technique that has been applied to study biological specimens ranging from individual proteins to cells, tissues, organelles, and even whole organisms. This protocol focuses on two chemical drying methods, hexamethyldisilazane (HMDS) and t-butyl alcohol (TBA), and their application to imaging of both prokaryotic and eukaryotic organisms using SEM. In this article, we describe how to fix, wash, dehydrate, dry, mount, sputter coat, and image three types of organisms: cyanobacteria (Toxifilum mysidocida, Golenkina sp., and an unknown sp.), two euglenoids from the genus Monomorphina (M. aenigmatica and M. pseudopyrum), and the fruit fly (Drosophila melanogaster). The purpose of this protocol is to describe a fast, inexpensive, and simple method to obtain detailed information about the structure, size, and surface characteristics of specimens that can be broadly applied to a large range of organisms for morphological assessment. Successful completion of this protocol will allow others to use SEM to visualize samples by applying these techniques to their system.

Preparation of Prokaryotic and Eukaryotic Organisms Using Chemical Drying for Morphological Analysis in Scanning Electron Microscopy SEM

SEM analysis is an effective method to aid in species identification or phenotypic discrimination. This protocol describes the methods for examining specific morphological details of three representative types of organisms and would be broadly applicable to examining features of many organismal and tissue types.

原核生物と真核生物の化学形態分析走査電子顕微鏡 (SEM) での乾燥の準備

Scanning electron microscopy (SEM) is a widely available technique that has been applied to study biological specimens ranging from individual proteins to ...

生物学

ナノハーブコーティング生地の抗菌効果

An Antimicrobial Fabric Using Nano-Herbal Encapsulation of Essential Oils

Lab coats are widely used in biohazard laboratories and healthcare facilities as protective garments to prevent direct exposure to pathogens, spills, and burns. These cotton-based protective coats provide ideal conditions for microbial growth and attachment sites due to their porous nature, moisture-holding capacity, and retention of warmth from the user&#39;s body. Several studies have demonstrated the survival of pathogenic bacteria on hospital garments and lab coats, acting as vectors of microbial transmission.
A common approach to fix these problems is the application of antimicrobial agents in textile finishing, but concerns have been raised due to the toxicity and environmental effects of many synthetic chemicals. The ongoing pandemic has also opened a window for the investigation of effective antimicrobials and eco-friendly and toxic-free formulations. This study uses two natural bioactive compounds, carvacrol and thymol, encapsulated in chitosan nanoparticles, which guarantee effective protection against four human pathogens with up to a 4-log reduction (99.99%). These pathogens are frequently detected in lab coats used in biohazard laboratories.
The treated fabrics also resisted up to 10 wash cycles with 90% microbial reduction, which is sufficient for the intended use. We made modifications to the existing standard fabric tests to better represent the typical scenarios of lab coat usage. These refinements allow for a more accurate evaluation of the effectiveness of antimicrobial lab coats and for the simulation of the fate of any accidental microbial spills that must be neutralized within a short time. Further studies are recommended to investigate the accumulation of pathogens over time on antimicrobial lab coats compared to regular protective coats.

著者スポットライト:エッセンシャルオイルのナノハーブカプセル化を使用した抗菌生地  

Antimicrobial lab coats prevent the cross-contamination of pathogen accumulation and accidental bio-spills. Here, we describe the protocol for developing a skin-friendly antimicrobial fabric using nano-herbal encapsulation and modified standard tests to precisely evaluate the efficacy and suitability for typical usage of the lab coat.

エッセンシャルオイルのナノハーブカプセル化を使用した抗菌ファブリック

Lab coats are widely used in biohazard laboratories and healthcare facilities as protective garments to prevent direct exposure to pathogens, spills, and ...

タイムのエッセンシャルオイルのアプリケーションの肉乾燥中の微生物負荷に及ぼす影響

要約

さらに動画を探す

この動画の章

定量的なアプリケーションのための均質なMALDI試料の調製

のサンプル調製

バイオコントロール製品の揮発性および不揮発性抗真菌活性の測定

イメージング質量分析による寒天上および感染組織内で増殖した細菌コロニーの微生物連携の検討

犬の口臭の管理のための栄養補助食品の治療有効性

等熱マイクロカロリメトリーを用いた新天然物の殺菌効果または静電効果を測定する代謝プロファイリング

結核分子細菌負荷アッセイ(TB-MBLA)

原核生物と真核生物の化学形態分析走査電子顕微鏡 (SEM) での乾燥の準備

エッセンシャルオイルのナノハーブカプセル化を使用した抗菌ファブリック

エッセンシャルオイルのナノハーブカプセル化を使用した抗菌ファブリック

タイムのエッセンシャル オイルのアプリケーションの肉乾燥中の微生物負荷に及ぼす影響

要約

さらに動画を探す

この動画の章

定量的なアプリケーションのための均質なMALDI試料の調製

のサンプル調製

バイオコントロール製品の揮発性および不揮発性抗真菌活性の測定

イメージング質量分析 による 寒天上および感染組織内で増殖した細菌コロニーの微生物連携の検討

犬の口臭の管理のための栄養補助食品の治療有効性

等熱マイクロカロリメトリーを用いた新天然物の殺菌効果または静電効果を測定する代謝プロファイリング

結核分子細菌負荷アッセイ(TB-MBLA)

原核生物と真核生物の化学形態分析走査電子顕微鏡 (SEM) での乾燥の準備

エッセンシャルオイルのナノハーブカプセル化を使用した抗菌ファブリック

エッセンシャルオイルのナノハーブカプセル化を使用した抗菌ファブリック

タイムのエッセンシャルオイルのアプリケーションの肉乾燥中の微生物負荷に及ぼす影響

イメージング質量分析による寒天上および感染組織内で増殖した細菌コロニーの微生物連携の検討