Yazarlar bu çalışma ile yardımcı gıda güvenliği laboratuvar öğrenciler ve gönüllüler teşekkür etmek istiyorum. Bu araştırma USDA Ulusal Entegre Gıda Güvenliği Girişimi (NIFSI) hibe (Ödül # 10507316) tarafından finanse edilmiştir.

1.. Standart Eğri <ol><li> 37.5 mg floresan toz tartılır ve 6 ml 200 proof etanol içinde çözülür. 200 derece etanol içindeki Bu çözeltiye 2-kat seri dilüsyonları olun. </li><li> Vortex tüm 10 saniye boyunca numune ve 370 nm&#39;de absorbans ölçümü. </li><li> 1 ml 200 derece etanol başına floresan tozu miktarını hesaplayın. </li><li> Floresan toz konsantrasyonuna göre absorbansı göstermek için (x-ekseni üzerinde 370 nm&#39;de y-ekseni ve emilime mg / ml konsantrasyon) standart bir eğri hazırlanır. </li></ol> 2. Floresan Tozu ile Deli Et (Bologna) inoküle <ol><li> Temiz bir bıçak kullanarak ve kesme tahtası yaklaşık 100 mm kalınlığındaki numuneler içine deli et kesti. A, B ve C işaretli üç numune hazırlayın </li><li> Coat örnek bir hafif nemli, temiz bir sünger kullanarak floresan tozu ile eşit bir. Eşit et numunesi üzerinde toz yayıldı. Floresan tozu ile B ve C numuneleri aşılamak etmeyin. </li&gt; <li> Plastik wrap ile örnekleri sarın ve &#39;A&#39; ve bant ile B ve C gibi iki numune kalan olarak floresan toz kaplanmış numunenin etiket. </li><li> Deli durum ya da deney gerçekleştirilecektir sahte mutfakta 4 ° C&#39;de buzdolabında tutulan numuneler. </li></ol> 3. Mock Deli Mutfak Hazırlık <ol><li> Mutfakta temiz eldiven ve% 70 etanol sprey sağlayın. </li><li> Dilimleyici alanı çevresinde 13 W ampul ile siyah kompakt floresan ışıkları monte edin. </li><li> Paspas için şablon olarak hizmet etmek üzere bir alüminyum folyo, 5 cm x 5 cm kesme hazırlayın. </li><li> 6 ml% 95 etanol ihtiva eden 15 ml tüpler hazırlayın. </li></ol> 4. Video Ayarı Aynı zamanda alay deli tüm alanları gözlemlemek için stratejik yerlerde Dağı üç video kameralar. (Protokol 5&#39;te açıklandığı gibi) dilimleme işlemi simüle önce kameralar açın ve gibi (eşzamanlı analiz sonuçları) Protokol 7 olarak açıkladı. 5. Çapraz Bulaşma Track Deli Dilimleme Prosedürleri benzetme Örneklem büyüklüğü artırmak ve varyasyon dahil, ilgili kurumun Kurumsal Değerlendirme Kurulu tarafından protokolün onayından sonra işe katılımcılar tarafından bu çalışmayı gerçekleştirmek için. Katılımcı ya da deneyim perakende deli ortamlarda çalışan yok olabilir bir üniversite öğrencisi olabilir. Onlar deli girdikten sonra her bir katılımcı eldiven giyer emin olun. Take a siyah bir floresan ışığı altında resim izledi &quot;önce&quot; bir çalışmanın tamamlanmasıyla resim &quot;sonra&quot;. Aşağıdaki gibi her katılımcıya açık yazılı ve sözlü talimatları sağlayın: <ol><li> Buzdolabına git. </li><li> &quot;A&quot; etiketli et çıkarın. </li><li> Et açınız ve plastik wrap kaydedin. </li><li> Dilimleyicinin taşıma tepsi üzerine eti yerleştirin. </li><li> Et kavrama ile et sabitleyin. </li><li> Güç anahtarı AÇIK konuma getirin. </li><li> SLI ayarlayıncer endeksi topuzu &quot;2&quot;. </li><li> Dilim ve şarküteri kağıda et 5 adet dağıtmak. </li><li> OFF güç anahtarını açın ve et kavrama bırakın. </li><li> &quot;A&quot; etiketli plastik torba içinde et dilimlerini yerleştirin. </li><li> Eti &quot;A&quot; sarmaktır ve buzdolabı dönmek. </li><li> TEKRAR &quot;B&quot; ve &quot;C&quot; etiketli etler ile 2-11 Adımları. </li></ol> 6. Miktarının Floresan Tozu Deli mutfakta sahte dilimleme (Bölüm 5.4) tamamlanmasından sonra floresan tozu niceliğini. Bu aşağıdakileri yapın için: <ol><li> Steril 5 cm x 5 cm alüminyum folyo şablonunu kullanarak ve Şekil 1 &#39;de gösterilen alanlar üzerine yerleştirin. </li><li> Bölgeyi çubukla,% 95 etanol ile ıslatılmış steril bir kalsiyum aljinat bezi kullanın ve steril bir polipropilen tüp içinde 6 ml% 95 etanol içinde yer. </li><li> Vorteks tüp iyice ve 370 nm&#39;de absorbans okumak için bir cam küvet transfer. </li><li> Standar bakınAşağıdaki denklem kullanılarak floresan toz miktarının (y) saptamak için eğri d: x emme y = mx + b, m, standart eğrinin eğimi ve b bölümüdür. </li><li> Verilerin normalleştirilmesi için, aşağıdaki formül kullanılarak örnek alan başına floresan tozun yüzde hesaplanır: </li></ol><img src="/files/ftp_upload/51402/51402eq1.jpg" alt="Formül 1" /> 7. Video Analiz Videotape katılımcılar tam görevleri 5,1-5,12 gibi. Sync ve tüm açıları aynı anda görülebilir, böylece DVD diskler için video kayıtları kaydedin. El dokunur sayısını deli ortamında (4 farklı araştırmacılar bağımsız seti komple verileri analiz edecek) çeşitli yüzeylerde (gönüllü alay deli alanlarına dokunmayın kez sayı) kaydetmek için videoları izleyin.

<ol>
	<li>Sheen, S., Hwang, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mathematical+modeling+the+cross-contamination+of+Escherichia+coli+O157:+H7+on+the+surface+of+ready-to-eat+meat+product+while+slicing.">Mathematical modeling the cross-contamination of Escherichia coli O157: H7 on the surface of ready-to-eat meat product while slicing.</a> Food Microbiol. 27, 37-43 (2010).</li><li>Crandall, P., Neal, 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=Minimizing+the+risk+of+Listeria+monocytogenes+in+retail+delis+by+developing+employee+focused,+cost+effective+training.">Minimizing the risk of Listeria monocytogenes in retail delis by developing employee focused, cost effective training.</a> Agric. Food Anal. Bacteriol. 1, 159-174 (2011).</li><li>Koo, O. K., Sirsat, S. 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=Physical+and+chemical+control+of+Salmonella+in+ready-to-eat+products.">Physical and chemical control of Salmonella in ready-to-eat products.</a> Agric. Food Anal. Bacteriol. 2, 56-68 (2012).</li><li>Hanning, I. B., Johnson, M. G., Ricke, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Precut+prepackaged+lettuce:+a+risk+for+listeriosis?.">Precut prepackaged lettuce: a risk for listeriosis?.</a> Foodborne Pathog. Dis. 5, 731-746 (2008).</li><li>Hanning, I. B., Nutt, J. D., Ricke, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Salmonellosis+outbreaks+in+the+United+States+due+to+fresh+produce:+sources+and+potential+intervention+measures.">Salmonellosis outbreaks in the United States due to fresh produce: sources and potential intervention measures.</a> Foodborne Pathog. Dis. 6, 635-648 (2008).</li><li>Zhu, M., Du, 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=Control+of+Listeria+monocytogenes+contamination+in+ready&#8208;to&#8208;eat+meat+products.">Control of Listeria monocytogenes contamination in ready&#8208;to&#8208;eat meat products.</a> Compr. Rev. Food Sci. Food Safety. 4, 34-42 (2005).</li><li>Lianou, A., Sofos, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+the+incidence+and+transmission+of+Listeria+monocytogenes+in+ready-to-eat+products+in+retail+and+food+service+environments.">A review of the incidence and transmission of Listeria monocytogenes in ready-to-eat products in retail and food service environments.</a> J. Food Protect. 70, 2172-2198 (2007).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+risk+assessment+of+relative+risk+to+public+health+from+foodborne+Listeria+monocytogenes+among+selected+categories+of+ready-to-eat+foods.">Quantitative risk assessment of relative risk to public health from foodborne Listeria monocytogenes among selected categories of ready-to-eat foods.</a> , .</li><li>Endrikat, S., Gallagher, D., 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=A+comparative+risk+assessment+for+Listeria+monocytogenes+in+prepackaged+versus+retail-sliced+deli+meat.">A comparative risk assessment for Listeria monocytogenes in prepackaged versus retail-sliced deli meat.</a> J. Food Protect. 73, 612-619 (2010).</li><li>Pradhan, A. K., Ivanek, 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=Comparison+of+public+health+impact+of+Listeria+monocytogenes+product-to-product+and+environment-to-product+contamination+of+deli+meats+at+retail.">Comparison of public health impact of Listeria monocytogenes product-to-product and environment-to-product contamination of deli meats at retail.</a> J. Food Protect. 74, 1860-1868 (2011).</li><li>Jones, C. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Handwashing+Technique+Analysis.">Handwashing Technique Analysis.</a> Patent number 5900067. , (1999).</li><li>Gibson, K. E., Koo, O. K., 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=Observation+and+relative+quantification+of+cross-contamination+within+a+mock+retail+delicatessen+environment.">Observation and relative quantification of cross-contamination within a mock retail delicatessen environment.</a> Food Control. 31, 116-124 (2013).</li><li>Xiao, M., Zheng, 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=Method+and+System+for+Fecal+Detection.">Method and System for Fecal Detection.</a> Patent number 5821546. , (1998).</li><li>Waldroup, A., Kirby, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Method+and+System+for+Fecal+Detection.">Method and System for Fecal Detection.</a> Patent number 5621215. , (1997).</li><li>Ashby, K. D., Wen, 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=Fluorescence+of+dietary+porphyrins+as+a+basis+for+real-time+detection+of+fecal+contamination+on+meat.">Fluorescence of dietary porphyrins as a basis for real-time detection of fecal contamination on meat.</a> J. Agric. Food Chem. 51, 3502-3507 (2003).</li><li>Cabrera-Diaz, E., Moseley, T. 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=Fluorescent+protein+marked+Escherichia+coli+biotype+I+strains+as+surrogates+for+enteric+pathogens+in+validation+of+beef+carcass+interventions.">Fluorescent protein marked Escherichia coli biotype I strains as surrogates for enteric pathogens in validation of beef carcass interventions.</a> J. Food Protect. 72, 295-303 (2009).</li><li>Byrd, J., Hargis, B., 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=Fluorescent+marker+for+the+detection+of+crop+and+upper+gastrointestinal+leakage+in+poultry+processing+plants.">Fluorescent marker for the detection of crop and upper gastrointestinal leakage in poultry processing plants.</a> Poult. Sci. 81, 70-74 (2002).</li><li>Vorst, K. L., Todd, E. C., 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=Transfer+of+Listeria+monocytogenes+during+mechanical+slicing+of+turkey+breast+bologna,+and+salami..">Transfer of Listeria monocytogenes during mechanical slicing of turkey breast bologna, and salami..</a> J. Food Protect. 69, 619-626 (2006).</li></ol>

Çıkar çatışması ilan etti.

Bu çalışmanın genel amacı bakteriyel çapraz kontaminasyon gıda kaynaklı hastalık riskini kontrol etmek ve azaltmak için stratejiler geliştirmek amacıyla bir perakende deli ortamda nasıl meydana gelebileceğini göstermekti. Perakende deli çevre L. için özellikle yüksek bir risk vardır kontaminasyonu monocytogenes. USDA-GGBS perakende mağazasında dilimlenmiş deli et deli et dilimlenmiş ve bir federal teftiş fabrikasında 9 paketlendiği listeriosisde ile yedi kat daha y...

Özellikle bireysel bölümündeki yiyecek hazırlama ortamında kaynaklı patojenlerin, çapraz kontaminasyon nedeniyle etler gibi sebze 1-5 de dahil olmak üzere çeşitli kaynaklardan gelen gıda kaynaklı bir hastalık riski için önemli bir husustur. Çoğu zaman, bakteriyel patojenler kirlenmiş gıda ürünleri ile 6 perakende ortamını girin. Tüketilmeden önce 7 başka müdahale veya tedavi (yani pişirme veya ısıtma), genellikle olduğu gibi perakende seviyesinde hazırlanmış hazır yiyecekleri özellikle endişe vardır. Ayrıca, kontamine RTE gıda mevcut patojenler daha sonra perakende ortamında diğer gıda ürünleri veya gıda ile temas eden yüzeylere transfer edilebilir. Çapraz kontaminasyon perakende gıda ortamında hemen hemen her tür içinde ortaya çıkabilir, delis nedeniyle özellikle patojen Listeria monocytog ile RTE şarküteri etler dernek özellikle ilgienes 2. ABD Gıda ve İlaç İdaresi (FDA) ve Tarım Gıda Güvenliği ve Denetim Servisi (USDA-GGBS) ABD Bakanlığı tarafından yapılan risk değerlendirmesi analizleri dayanarak, kirlenmiş şarküteri listeryoz vakaların% 90 olasılıkla sorumludur - nadir bir L. neden henüz ciddi insan hastalığı monocytogenes - Amerika Birleşik Devletleri&#39;nde 8. Genel olarak, gıda ve perakende işyerlerinde hizmet dilimlenmiş şarküteri L. riski ile ilişkili bulunmuştur monocytogenes üretici 9 ile USDA denetlenen fabrikasında presliced ​​şarküteri göre. Bunun nedeni gıda işleyicileri, gelen ham maddeler, ya da tedavi sonrası 7 kirlenmiş olabilir işlenmiş gıdalar patojenlerin giriş ihtimalinin artması olabilir. Genel olarak, listeriozis ABD&#39;de yılda gıda kaynaklı hastalığa neden tahmin ölümlerin yaklaşık% 28&#39;ini Çünkü InL. artmış risk perakende seviyesinde dilimlenmiş şarküteri etler kontaminasyonu monocytogenes, deli dilimleme kendisi perakende ortamında 2,10 gıda kaynaklı patojenlerin yüzey transferi için önemli bir faktör olarak tespit edilmiştir. Sheen 1 olarak gösterilen, dilimleme tüketilmeden önce son işlem adımdır, bu nedenle deli dilimleme anlayış ve çapraz bulaşmayı 1 önlenmesi hem bir kritik kontrol noktası olarak kabul edilmelidir. Bu çalışmanın genel amacı gibi Glo Germ gibi bir floresan tozu kullanılarak perakende deli ortamında kirlenmeyi izlemek için bir metodoloji geliştirmek oldu. Bu bakteri çapraz bulaşmayı ve sıhhi uygulamaları 11,12 simüle etmek için kullanılan bir floresan bileşiktir. Bu çalışma dilimleme, çevresini ve diğer şarküteri kontamine etler patojen transferi izlemek için tasarlanmıştır. Bu cont ve yolları anlamak için çok önemlidiretkili girişim ve perakende çalışanlar ve yöneticiler için eğitim materyali hem de geliştirmek için aminleştirme. Benzer bir yaklaşım, diğer işleme ortamlarda (örneğin tavuk ve sığır eti), çiftlikler ve hastanelerde patojen aktarımını simüle etmek için kullanılabilir.

<table border="0" cellpadding="0" cellspacing="0" width="637">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Glo-Germ Powder</td>
			<td style="width:71px;">Glo Germ Co</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">Purchased from vendor</td>
			<td style="width:64px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Ethanol</td>
			<td style="width:71px;">Sigma</td>
			<td style="width:119px;">E7023</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Permanent markets</td>
			<td style="width:71px;">Sharpie</td>
			<td style="width:119px;"></td>
			<td colspan="2">Purchased from stationary store</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Gloves</td>
			<td style="width:71px;">VWR</td>
			<td style="width:119px;">82026-424</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Deli Meat</td>
			<td style="width:71px;">NA</td>
			<td style="width:119px;">NA</td>
			<td colspan="2">Bologna Chub from regular grocery store</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Cutting Board</td>
			<td style="width:71px;">NA</td>
			<td style="width:119px;">NA</td>
			<td colspan="2">A regukar kitchen cutting board</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Knife</td>
			<td style="width:71px;">NA</td>
			<td style="width:119px;">NA</td>
			<td>A regular kitchen knife&#160;</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">5 x 5 cm sterile templates</td>
			<td style="width:71px;">NA</td>
			<td style="width:119px;">NA</td>
			<td colspan="2">Aluminium foil templates cut into 5x 5 cm templates and sterilized&#160;</td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">15 ml polypropylene centrifuge tubes</td>
			<td style="width:71px;">VWR</td>
			<td style="width:119px;">89039-664</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Cotton swabs</td>
			<td style="width:71px;">Puritan</td>
			<td style="width:119px;">25-806</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Glass cuvettes</td>
			<td style="width:71px;">VWR</td>
			<td style="width:119px;">470019-186</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Vortex</td>
			<td style="width:71px;">VWR</td>
			<td style="width:119px;">58816-121</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Flip camera</td>
			<td style="width:71px;">Flip Ultra HD</td>
			<td style="width:119px;">NA</td>
			<td>Purchased online</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Deli slicer</td>
			<td style="width:71px;">Bizerba</td>
			<td style="width:119px;">SE-12</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="43">
			<td height="43" style="height:43px;width:216px;">Deli refrigerator</td>
			<td style="width:71px;">True company</td>
			<td style="width:119px;">TDBD-722</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:216px;">Scale</td>
			<td style="width:71px;"></td>
			<td style="width:119px;">NA</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="85">
			<td height="85" style="height:85px;width:216px;">Spectrophotometer</td>
			<td style="width:71px;">Milton Roy Spectronic 20D</td>
			<td style="width:119px;">NA</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

tracking microbial contamination in retail environments using fluorescent powder - a retail delicatessen environment example

Perakende ortamında gıda kaynaklı patojenlerin çapraz kontaminasyon gıda kaynaklı hastalık riskinin artmasına katkıda önemli bir halk sağlığı sorunudur. Böyle deli et, peynir, ve bazı durumlarda taze üretmek gibi hazır yemek (RTE) işlenmiş gıdalar gibi Listeria monocytogenes gibi patojen bulaşma nedeniyle gıda kaynaklı hastalık salgınları dahil edilmiştir. L. ile ilgili olarak monocytogenes, deli dilimleme genellikle çapraz bulaşma ana kaynağıdır. Bu çalışmanın amacı bakteri kontaminasyonu taklit ve bir perakende ortamında bu kirlenme izlemek için floresan bir bileşiği kullanmak oldu. Bir alay deli mutfak perakende ortamını simüle etmek için tasarlanmıştır. Deli et floresan bileşik ile aşılanır ve gönüllü bir gıda perakende çalışanın beklenen benzer görevleri bir dizi tamamlamak için işe alındı. Gönüllüler, paket dilim ve bir deli buzdolabında et saklamak için talimatı verildi. Potansiyelbulaşma belirli alanlarda swabbing ve bir spektrofotometre ile temizlenmesi alanının optik yoğunluğu ölçerek sahte perakende ortamında takip edildi. Sonuçlar dilimleyicide buzdolabı (yani deli durumda) kavrama ve çeşitli alanlarda çapraz kontaminasyon riskinin en yüksek olduğunu gösterdi. Bu çalışmanın sonuçları, perakende çalışanlar için daha odaklı eğitim materyali geliştirmek için kullanılabilir. Buna ek olarak, benzer metodolojileri de gıda üretim ortamlarında (örneğin küçük çiftlikler), hastaneler, bakım evleri, yolcu gemileri ve otellerde mikrobiyal kontaminasyonu izlemek için kullanılabilir.

Şekil 1 gönüllüler atanan görevleri tamamlandıktan sonra silinmiştir dilimleyicinin alanlarını temsil eder. Gönüllüler bu çeşitli yüzeylerde el temas ortalama sıklığı analiz videoya alındı. El temas sıklığı dört farklı gözlemciler tarafından analiz edilmiş ve ortalama alındı. Bu sonuçlar, Şekil 2&#39;de gösterilmiştir. Veri et kavrama, deli et sarıcı, deli et ve şarküteri kağıt el teması (8-14 kontakların ortalama) en yüksek oranda olduğunu ...

Watch this Scientific Journal Video about Tracking Microbial Contamination in Retail Environments Using Fluorescent Powder - A Retail Delicatessen Environment Example at JoVE.com

Tracking Microbial Contamination in Retail Environments Using Fluorescent Powder - A Retail Delicatessen Environment Example

Bu çalışmanın genel amacı, bir perakende deli bir ortamda gıda kaynaklı patojen Listeria monocytogenes potansiyel çapraz kontaminasyon mekanizmasını göstermekti. Bu metodoloji, patojen kirlenme izlemek için farklı ortamlarda çeşitli tatbik edilebilir.

Floresan Tozu Kullanımı Perakende Ortamlarda Mikrobiyal Bulaşma Takip - Bir Perakende Deniz Çevre Örneği

Cross contamination of foodborne pathogens in the retail environment is a significant public health issue contributing to an increased risk for foodborne ...

tracking-microbial-contamination-retail-environments-using

Research

JoVE Journal

Environment

11.0K Görüntüleme.  University of Houston.  Bu çalışmanın genel amacı, bir perakende deli bir ortamda gıda kaynaklı patojen Listeria monocytogenes potansiyel çapraz kontaminasyon mekanizmasını göstermekti. Bu metodoloji, patojen kirlenme izlemek için farklı ortamlarda çeşitli tatbik edilebilir. 

Video: Tracking Microbial Contamination in Retail Environments Using Fluorescent Powder - A Retail Delicatessen Environment Example

Determination of Microbial Extracellular Enzyme Activity in Waters, Soils, and Sediments using High Throughput Microplate Assays

Much of the nutrient cycling and carbon processing in natural environments occurs through the activity of extracellular enzymes released by microorganisms. Thus, measurement of the activity of these extracellular enzymes can give insights into the rates of ecosystem level processes, such as organic matter decomposition or nitrogen and phosphorus mineralization. Assays of extracellular enzyme activity in environmental samples typically involve exposing the samples to artificial colorimetric or fluorometric substrates and tracking the rate of substrate hydrolysis. Here we describe microplate based methods for these procedures that allow the analysis of large numbers of samples within a short time frame. Samples are allowed to react with artificial substrates within 96-well microplates or deep well microplate blocks, and enzyme activity is subsequently determined by absorption or fluorescence of the resulting end product using a typical microplate reader or fluorometer. Such high throughput procedures not only facilitate comparisons between spatially separate sites or ecosystems, but also substantially reduce the cost of such assays by reducing overall reagent volumes needed per sample.

Microplate based procedures are described for the colorimetric or fluorometric analysis of extracellular enzyme activity. These procedures allow for the rapid assay of such activity in large numbers of environmental samples within a manageable time frame.

Yüksek Verimli Mikroplate Tahliller kullanarak Waters, Topraklar ve tortulları Mikrobiyal Ekstrasellüler Enzim Aktivitesi Belirlenmesi

Much of the nutrient cycling and carbon  processing in natural environments occurs through the activity of extracellular  enzymes released by ...

Çevre

Technique for Studying Arthropod and Microbial Communities within Tree Tissues

Phloem tissues of pine are habitats for many thousands of organisms. Arthropods and microbes use phloem and cambium tissues to seek mates, lay eggs, rear young, feed, or hide from natural enemies or harsh environmental conditions outside of the tree. Organisms that persist within the phloem habitat are difficult to observe given their location under bark. We provide a technique to preserve intact phloem and prepare it for experimentation with invertebrates and microorganisms. The apparatus is called a &#8216;phloem sandwich&#8217; and allows for the introduction and observation of arthropods, microbes, and other organisms. This technique has resulted in a better understanding of the feeding behaviors, life-history traits, reproduction, development, and interactions of organisms within tree phloem. The strengths of this technique include the use of inexpensive materials, variability in sandwich size, flexibility to re-open the sandwich or introduce multiple organisms through drilled holes, and the preservation and maintenance of phloem integrity. The phloem sandwich is an excellent educational tool for scientific discovery in both K-12 science courses and university research laboratories.

We provide a technique to preserve intact tree phloem and prepare it for observation. We create an apparatus called a phloem sandwich that allows for the introduction and observation of arthropods, microbes, and other organisms that inhabit phloem tissues.

Ağaç Dokuların içinde eklembacaklı ve Mikrobiyal Toplulukları incelenmesi için Tekniği

Phloem tissues of pine are habitats for many thousands of organisms.  Arthropods and microbes use phloem and cambium tissues to seek mates, lay eggs, rear ...

An Aquatic Microbial Metaproteomics Workflow: From Cells to Tryptic Peptides Suitable for Tandem Mass Spectrometry-based Analysis

Meta-omic technologies such as metagenomics, metatranscriptomics and metaproteomics can aid in the understanding of microbial community structure and metabolism. Although powerful, metagenomics alone can only elucidate functional potential. On the other hand, metaproteomics enables the description of the expressed in situ metabolism and function of a community. Here we describe a protocol for cell lysis, protein and DNA isolation, as well as peptide digestion and extraction from marine microbial cells collected on a cartridge filter unit (such as the Sterivex filter unit) and preserved in an RNA stabilization solution (like RNAlater). In mass spectrometry-based proteomics studies, the identification of peptides and proteins is performed by comparing peptide tandem mass spectra to a database of translated nucleotide sequences. Including the metagenome of a sample in the search database increases the number of peptides and proteins that can be identified from the mass spectra. Hence, in this protocol DNA is isolated from the same filter, which can be used subsequently for metagenomic analysis.

This protocol is for the extraction and concentration of protein and DNA from microbial biomass collected from seawater, followed by the generation of tryptic peptides suitable for tandem mass spectrometry-based proteomic analysis.

Bir Aquatic Mikrobiyal Metaproteomics İş Akışı: Tandem Kütle Spektrometre tabanlı analiz için uygun Triptik Peptitler için Hücreleri itibaren

Meta-omic technologies such as metagenomics, metatranscriptomics and metaproteomics can aid in the understanding of microbial community structure and ...

Extraction and Analysis of Microbial Phospholipid Fatty Acids in Soils

Phospholipid fatty acids (PLFAs) are key components of microbial cell membranes. The analysis of PLFAs extracted from soils can provide information about the overall structure of terrestrial microbial communities. PLFA profiling has been extensively used in a range of ecosystems as a biological index of overall soil quality, and as a quantitative indicator of soil response to land management and other environmental stressors.
The standard method presented here outlines four key steps: 1. lipid extraction from soil samples with a single-phase chloroform mixture, 2. fractionation using solid phase extraction columns to isolate phospholipids from other extracted lipids, 3. methanolysis of phospholipids to produce fatty acid methyl esters (FAMEs), and 4. FAME analysis by capillary gas chromatography using a flame ionization detector (GC-FID). Two standards are used, including 1,2-dinonadecanoyl-sn-glycero-3-phosphocholine (PC(19:0/19:0)) to assess the overall recovery of the extraction method, and methyl decanoate (MeC10:0) as an internal standard (ISTD) for the GC analysis.

Phospholipid fatty acids provide information about the structure of soil microbial communities. We present methods for extraction from soil samples with a single-phase chloroform mixture, fractionation of extracted lipids using solid phase extraction columns, and methanolysis to produce fatty acid methyl esters, which are analyzed by capillary gas chromatography.

Ekstraksiyon ve Topraklarda Mikrobiyal Fosfolipid Yağ Asitleri Analizi

Phospholipid fatty acids (PLFAs) are key components of microbial cell membranes. The analysis of PLFAs extracted from soils can provide information about ...

Sampling, Sorting, and Characterizing Microplastics in Aquatic Environments with High Suspended Sediment Loads and Large Floating Debris

The ubiquitous presence of plastic debris in the ocean is widely recognized by the public, scientific communities, and government agencies. However, only recently have microplastics in freshwater systems, such as rivers and lakes, been quantified. Microplastic sampling at the surface usually consists of deploying drift nets behind either a stationary or moving boat, which limits the sampling to environments with low levels of suspended sediments and floating or submerged debris. Previous studies that employed drift nets to collect microplastic debris typically used nets with &#8805;300 &#181;m mesh size, allowing plastic debris (particles and fibers) below this size to pass through the net and elude quantification. The protocol detailed here enables: 1) sample collection in environments with high suspended loads and floating or submerged debris and 2) the capture and quantification of microplastic particles and fibers &lt;300 &#181;m. Water samples were collected using a peristaltic pump in low-density polyethylene (PE) containers to be stored before filtering and analysis in the lab. Filtration was done with a custom-made microplastic filtration device containing detachable union joints that housed nylon mesh sieves and mixed cellulose ester membrane filters. Mesh sieves and membrane filters were examined with a stereomicroscope to quantify and separate microplastic particulates and fibers. These materials were then examined using a micro-attenuated total reflectance Fourier transform infrared spectrometer (micro ATR-FTIR) to determine microplastic polymer type. Recovery was measured by spiking samples using blue PE particulates and green nylon fibers; percent recovery was determined to be 100% for particulates and 92% for fibers. This protocol will guide similar studies on microplastics in high velocity rivers with high concentrations of sediment. With simple modifications to the peristaltic pump and filtration device, users can collect and analyze various sample volumes and particulate sizes.

Most microplastic research to date has occurred in marine systems where suspended solid levels are relatively low. Focus is now shifting to freshwater systems, which may feature high sediment loads and floating debris. This protocol addresses collecting and analyzing microplastic samples from aquatic environments that contain high suspended solid loads.

Örnekleme, sıralama ve Microplastics su ortamlarında yüksek askıya alınan tortu yükler ve büyük yüzen enkaz ile karakterize

The ubiquitous presence of plastic debris in the ocean is widely recognized by the public, scientific communities, and government agencies. However, only ...

Detection of 3-Nitrotyrosine in Atmospheric Environments via a High-performance Liquid Chromatography-electrochemical Detector System

3-nitrotyrosine (3-NT) is generated from the tyrosine residue in atmospheric bio-aerosol proteins via a reaction with ozone (O3) and nitrogen dioxide (NO2). Stable 3-NT is a specific marker for oxidative damage and is reported to have a promotive effect to elicit allergies. In the present study, we report the development of a highly sensitive assay to quantify 3-NT in air sampler filters to collect &#60; 2.5 &#181;m of particulate matter (PM2.5) from urban environmental air, including bio-aerosol. In this method, a 6 mm-diameter round hole was cut from the filters of air samplers and mixed with a nonspecific protease cocktail in order to hydrolyze proteins. Protein samples digested to the amino acid level were tested for 3-NT using a high-performance liquid chromatography-electrochemical detector (HPLC-ECD). The maximum 3-NT content was detected in a prefilter for PM of sizes from 4.5 to 7.3 &#956;m, with a detection limit of 1.13 pg/m3.

Detection of 3-Nitrotyrosine in Atmospheric Environments via a High-performance Liquid Chromatography-electrochemical Detector System

We present a method to detect 3-nitrotyrosine chemical modifications of atmospheric proteins with 6 mm-diameter rounds cut from air sampler prefilters using a high-performance liquid chromatography-electrochemical detector (HPLC-ECD).

3-Nitrotyrosine atmosferik ortamlar üzerinden yüksek performanslı sıvı kromatografi elektrokimyasal dedektör sistemi tespiti

3-nitrotyrosine (3-NT) is generated from the tyrosine residue in atmospheric bio-aerosol proteins via a reaction with ozone (O3) and nitrogen dioxide ...

Fluorescently Labeled Bacteria as a Tracer to Reveal Novel Pathways of Organic Carbon Flow in Aquatic Ecosystems

Elucidating trophic interactions, such as predation and its effects, is a frequent task for many researchers in ecology. The study of microbial communities has many limitations, and determining a predator, prey, and predatory rates is often difficult. Presented here is an optimized method based on the addition of fluorescently labelled prey as a tracer, which allows for reliable quantitation of the grazing rates in aquatic predatory eukaryotes and estimation of nutrient transfer to higher trophic levels.

Presented here is a protocol for a single-cell, epifluorescence microscopy-based technique to quantify grazing rates in aquatic predatory eukaryotes with high precision and taxonomic resolution.

Floresan Etiketli Bakteriler Sucul Ekosistemlerde Organik Karbon Akışının Yeni Yollarını Ortaya Çıkaracak Bir İzleyici Olarak

Elucidating trophic interactions, such as predation and its effects, is a frequent task for many researchers in ecology. The study of microbial ...

Enrichment and Detection of Clostridium perfringens Toxinotypes in Retail Food Samples

Clostridium perfringens (C. perfringens) is a prolific toxin producer and causes a wide range of diseases in various hosts. C. perfringens is categorized into five different toxinotypes, A through E, based on the carriage of four major toxin genes. The prevalence and distribution of these various toxinotypes is understudied, especially their pervasiveness in American retail food. Of particular interest to us are the type B and D strains, which produce epsilon toxin, an extremely lethal toxin suggested to be the environmental trigger of multiple sclerosis in humans. To evaluate the presence of different C. perfringens toxinotypes in various food samples, we developed an easy method to selectively culture these bacteria without the use of an anaerobic container system only involving three culturing steps. Food is purchased from local grocery stores and transported to the laboratory under ambient conditions. Samples are minced and inoculated into modified rapid perfringens media (RPM) and incubated overnight at 37 &#176;C in a sealed, airtight conical tube. Overnight cultures are inoculated onto a bottom layer of solid Tryptose Sulfite Cycloserine (TSC) agar, and then overlaid with a top layer of molten TSC agar, creating a &#34;sandwiched&#34;, anaerobic environment. Agar plates are incubated overnight at 37 &#176;C and then evaluated for appearance of black, sulfite-reducing colonies. C. perfringens-suspected colonies are removed from the TSC agar using sterile eye droppers, and inoculated into RPM and sub-cultured overnight at 37 &#176;C in an airtight conical tube. DNA is extracted from the RPM subculture, and then analyzed for the presence of C. perfringens toxin genes via polymerase chain reaction (PCR). Depending on the type of food sampled, typically 15&#8211;20% of samples test positive for C. perfringens.

Enrichment and Detection of Clostridium perfringens Toxinotypes in Retail Food Samples

The objective of this protocol is to detect different Clostridium perfringens toxinotypes in locally purchased foods, particularly epsilon toxin producing strain types B and D, without the use of anaerobic chambers.

Perakende Gıda Numunelerinde Clostridium Perfringens Toxinotypes Zenginleştirme ve Algılama

Clostridium perfringens (C. perfringens) is a prolific toxin producer and causes a wide range of diseases in various hosts. C. perfringens is categorized ...

In Situ Chemotaxis Assay to Examine Microbial Behavior in Aquatic Ecosystems

Microbial behaviors, such as motility and chemotaxis (the ability of a cell to alter its movement in response to a chemical gradient), are widespread across the bacterial and archaeal domains. Chemotaxis can result in substantial resource acquisition advantages in heterogeneous environments. It also plays a crucial role in symbiotic interactions, disease, and global processes, such as biogeochemical cycling. However, current techniques restrict chemotaxis research to the laboratory and are not easily applicable in the field. Presented here is a step-by-step protocol for the deployment of the in situ chemotaxis assay (ISCA), a device that enables robust interrogation of microbial chemotaxis directly in the natural environment. The ISCA is a microfluidic device consisting of a 20 well array, in which chemicals of interest can be loaded. Once deployed in aqueous environments, chemicals diffuse out of the wells, creating concentration gradients that microbes sense and respond to by swimming into the wells via chemotaxis. The well contents can then be sampled and used to (1) quantify strength of the chemotactic responses to specific compounds through flow cytometry, (2) isolate and culture responsive microorganisms, and (3) characterize the identity and genomic potential of the responding populations through molecular techniques. The ISCA is a flexible platform that can be deployed in any system with an aqueous phase, including marine, freshwater, and soil environments.

Presented here is the protocol for an in situ chemotaxis assay, a recently developed microfluidic device that enables studies of microbial behavior directly in the environment.

Sucul Ekosistemlerde Mikrobiyal Davranışı İncelemek Için Situ Chemotaxis Tsay'da

Microbial behaviors, such as motility and chemotaxis (the ability of a cell to alter its movement in response to a chemical gradient), are widespread ...

Evaluating the Impact of Hydraulic Fracturing on Streams using Microbial Molecular Signatures

Hydraulic fracturing (HF), commonly called &#34;fracking&#34;, uses a mixture of high-pressure water, sand, and chemicals to fracture rocks, releasing oil and gas. This process revolutionized the U.S. energy industry, as it gives access to resources that were previously unobtainable and now produces two-thirds of the total natural gas in the United States. Although fracking has had a positive impact on the U.S. economy, several studies have highlighted its detrimental environmental effects. Of particular concern is the effect of fracking on headwater streams, which are especially important due to their disproportionately large impact on the health of the entire watershed. The bacteria within those streams can be used as indicators of stream health, as the bacteria present and their abundance in a disturbed stream would be expected to differ from those in an otherwise comparable but undisturbed stream. Therefore, this protocol aims to use the bacterial community to determine if streams have been impacted by fracking. To this end, sediment, and water samples, from streams near fracking (potentially impacted) and upstream or in a different watershed of fracking activity (unimpacted) must be collected. Those samples are then subjected to nucleic acid extraction, library preparation, and sequencing to investigate microbial community composition. Correlational analysis and machine learning models can subsequently be employed to identify which features are explanative of variation in the community, as well as identification of predictive biomarkers for fracking&#39;s impact. These methods can reveal a variety of differences in the microbial communities among headwater streams, based on the proximity to fracking, and serve as a foundation for future investigations on the environmental impact of fracking activities.

Here, we present a protocol to investigate the impacts of hydraulic fracturing on nearby streams by analyzing their water and sediment microbial communities.

Hidrolik Çatlatmanın Akışlar Üzerindeki Etkisini Mikrobiyal Moleküler İmzalar Kullanarak Değerlendirmek

Hydraulic fracturing (HF), commonly called &#34;fracking&#34;, uses a mixture of high-pressure water, sand, and chemicals to fracture rocks, releasing oil ...