Microbial enrichment and multiplexed microfiltration for accelerated detection of Salmonella in spinach

Ku Seockmo; Ximenes Eduardo; Kreke Thomas; Foster Kirk; Couetil Justin L.; Zuponcic Jessica; Zhao Xiaojun; Hoagland Lori; Deering Amanda J.; Ladisch Michael R.

摘要

Abstract > To attain Salmonella detection thresholds in spinach suspensions using enrichment media requires at least 24?hr. Separation and concentration of selected microorganisms via microfiltration and microfugation reduce time for sample preparation, especially when working with large volumes of vegetable suspensions. This facilitates accelerated detection of Salmonella in spinach suspensions, and may contribute to effectively monitoring this pathogen before it reaches the consumer. We report a microfiltration‐based protocol for accelerated sample preparation to concentrate and recover ≤1 colony forming unit (CFU) Salmonella /g pathogen‐free spinach. Store‐bought samples of spinach and a spinach plant subjected to two environmental conditions (temperature and light exposure) during its production were tested. The overall procedure involves extraction with buffer, a short enrichment step, prefiltration using a nylon filter, crossflow hollow fiber microfiltration, and retentate centrifugation to bring microbial cells to detection levels. Based on 1 CFU Salmonella /g frozen spinach, and a Poisson distribution statistical analyses with 99% probability, we calculated that 3 hr of incubation, when followed by microfiltration, is sufficient to reach the 2 log concentration required for Salmonella detection within 7 hr. Longer enrichment times (5 hr or more) is needed for concentrations lower than 1 CFU Salmonella /g of ready to eat spinach. The recovered microbial cells were identified and confirmed as Salmonella using both polymerase chain reaction (PCR) and plating methods. Different environmental conditions tested during production did not affect Salmonella viability; this demonstrated the broad adaptability of Salmonella and emphasized the need for methods that enable efficient monitoring of production for the presence of this pathogen. </abstract> </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> <div class="translation abstracttxt"> <span class="zhankaihshouqi fivelineshidden" id="abstract"> <span>机译：</span><Abstract Type =“Main”XML：Lang =“en”> <标题类型=“main”>抽象</ title> > 达到沙门氏菌</ i> 使用富集介质的菠菜悬浮液中的检测阈值需要至少24个小时。通过微滤和微胶凝的选定微生物的分离和浓缩减少样品制备的时间，特别是在使用大量的植物悬浮液时。这有助于加速检测沙门氏菌</ i> 在菠菜悬浮液中，并且可能有助于在它到达消费者之前有效地监测该病原体。我们报告了一种基于微滤的方案，用于加速样品制剂以浓缩和回收≤1菌落形成单元（CFU）沙门氏菌</ i> / g无病原体菠菜。测试了菠菜的菠菜样品和经过两种环境条件（温度和曝光）的菠菜厂进行了测试。整体过程涉及用缓冲液提取，使用尼龙过滤器，交叉流中空纤维微滤，并保持离心，使微生物细胞带入检测水平。基于1 CFU 沙门氏菌</ i> / g冷冻菠菜，以及泊松分布统计分析，概率为99％，我们计算出3小时的孵育，当之后进行微滤，足以达到所需的2个日志浓度沙门氏菌</ i> 7小时内检测。浓度低于1 CFU的浓度需要更长的富集时间（5小时或更高）沙门氏菌</ i> / g准备吃菠菜。鉴定回收的微生物细胞并确认为沙门氏菌</ i> 使用聚合酶链反应（PCR）和电镀方法。生产过程中测试的不同环境条件不会影响沙门氏菌</ i> 可行性;这证明了广泛的适应性沙门氏菌</ i> 并强调需要能够有效监测生产该病原体的方法。 </ p> </摘要> </span> <span class="z_kbtn z_kbtnclass hoverxs" style="display: none;">展开▼</span> </div> </div> <div class="record"> <h2 class="all_title" id="enpatent33" >著录项</h2> <ul> <li> <span class="lefttit">来源</span> <div style="width: 86%;vertical-align: text-top;display: inline-block;"> <a href='/journal-foreign-15005/'>《Biotechnology Progress》</a> <b style="margin: 0 2px;">|</b><span>2019年第6期</span><b style="margin: 0 2px;">|</b><span>共10页</span> </div> </li> <li> <div class="author"> <span class="lefttit">作者</span> <p id="fAuthorthree" class="threelineshidden zhankaihshouqi"> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Ku Seockmo&option=202" target="_blank" rel="nofollow">Ku Seockmo;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Ximenes Eduardo&option=202" target="_blank" rel="nofollow">Ximenes Eduardo;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Kreke Thomas&option=202" target="_blank" rel="nofollow">Kreke Thomas;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Foster Kirk&option=202" target="_blank" rel="nofollow">Foster Kirk;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Couetil Justin L.&option=202" target="_blank" rel="nofollow">Couetil Justin L.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Zuponcic Jessica&option=202" target="_blank" rel="nofollow">Zuponcic Jessica;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Zhao Xiaojun&option=202" target="_blank" rel="nofollow">Zhao Xiaojun;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Hoagland Lori&option=202" target="_blank" rel="nofollow">Hoagland Lori;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Deering Amanda J.&option=202" target="_blank" rel="nofollow">Deering Amanda J.;</a> <a href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Ladisch Michael R.&option=202" target="_blank" rel="nofollow">Ladisch Michael R.;</a> </p> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zkzz" style="display: none;">展开▼</span> </div> </li> <li> <div style="display: flex;"> <span class="lefttit">作者单位</span> <div style="position: relative;margin-left: 3px;max-width: 639px;"> <div class="threelineshidden zhankaihshouqi" id="fOrgthree"> <p>Laboratory of Renewable Resources EngineeringPurdue UniversityWest Lafayette Indiana;</p> <p>Laboratory of Renewable Resources EngineeringPurdue UniversityWest Lafayette Indiana;</p> <p>Laboratory of Renewable Resources EngineeringPurdue UniversityWest Lafayette Indiana;</p> <p>Weldon School of Biomedical EngineeringPurdue UniversityWest Lafayette Indiana;</p> <p>Laboratory of Renewable Resources EngineeringPurdue UniversityWest Lafayette Indiana;</p> <p>Laboratory of Renewable Resources EngineeringPurdue UniversityWest Lafayette Indiana;</p> <p>Department of Horticulture and Landscape ArchitecturePurdue UniversityWest Lafayette Indiana;</p> <p>Department of Horticulture and Landscape ArchitecturePurdue UniversityWest Lafayette Indiana;</p> <p>Department of Food SciencePurdue UniversityWest Lafayette Indiana;</p> <p>Laboratory of Renewable Resources EngineeringPurdue UniversityWest Lafayette Indiana;</p> </div> <span class="z_kbtnclass z_kbtnclassall hoverxs" id="zhdw" style="display: none;">展开▼</span> </div> </div> </li> <li > <span class="lefttit">收录信息</span> <span style="width: 86%;vertical-align: text-top;display: inline-block;"></span> </li> <li> <span class="lefttit">原文格式</span> <span>PDF</span> </li> <li> <span class="lefttit">正文语种</span> <span>eng</span> </li> <li> <span class="lefttit">中图分类</span> <span><a href="https://www.zhangqiaokeyan.com/clc/15.html" title="生物科学">生物科学;</a></span> </li> <li class="antistop"> <span class="lefttit">关键词</span> <p style="width: 86%;vertical-align: text-top;"> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=detection&option=203" rel="nofollow">detection;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=microbial enrichment&option=203" rel="nofollow">microbial enrichment;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=multiplexed microfiltration&option=203" rel="nofollow">multiplexed microfiltration;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=Salmonella&option=203" rel="nofollow">Salmonella;</a> <a style="color: #3E7FEB;" href="/search.html?doctypes=4_5_6_1-0_4-0_1_2_3_7_9&sertext=spinach&option=203" rel="nofollow">spinach;</a> </p> <div class="translation"> 机译：检测;微生物富集;多重微滤;沙门氏菌;菠菜; </div> </li> </ul> </div> </div> <div class="literature cardcommon" id="literaturereference" style="display:none"> <div class="similarity "> <h3 class="all_title" id="enpatent111">引文网络</h3> <div class="referencetab clearfix"> <ul id="referencedaohang"> <li dataid="referenceul">参考文献</li> <li dataid="citationul">引证文献</li> <li dataid="commonreferenceul">共引文献</li> <li dataid="commoncitationul">同被引文献</li> <li dataid="tworeferenceul">二级参考文献</li> <li dataid="twocitationul">二级引证文献</li> </ul> </div> <div class="reference_details" id="referenceList"> <ul id="referenceul"></ul> <ul id="citationul"></ul> <ul id="commonreferenceul"></ul> <ul id="commoncitationul"></ul> <ul id="tworeferenceul"></ul> <ul id="twocitationul"></ul> </div> </div> </div> <div class="literature cardcommon"> <div class="similarity "> <h3 class="all_title" id="enpatent66">相似文献</h3> <div class="similaritytab clearfix"> <ul> <li class="active" >外文文献</li> <li >中文文献</li> <li >专利</li> </ul> </div> <div class="similarity_details"> <ul > <li> <div> <b>1. </b><a class="enjiyixqcontent" href="/journal-foreign-detail/0704021880122.html">Microbial enrichment and multiplexed microfiltration for accelerated detection of 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