Name: application of the intelligent high-throughput antimicrobial sensitivity testing/phage screening system and lar index of antimicrobial resistance
Uploaded: 2023-07-21T14:45:00
Description: Watch this Scientific Journal Video about Application of the Intelligent High-Throughput Antimicrobial Sensitivity Testing/Phage Screening System and Lar Index of Antimicrobial Resistance at JoVE.com

本工作得到国家重点研发专项（2019YFA0904003）资助;山东省现代农业产业体系（SDAIT-011-09）;国际合作平台优化项目（CXGC2023G15）;山东省农业科学院农业科技创新项目重大创新任务（CXGC2023G03）.

本研究中使用的 沙门氏菌菌 株是在获得中国山东省农业科学院畜牧兽医研究所生物安全委员会的批准后从中国山东家禽中采集的。
1. 智能高通量AST系统的应用8
<ol><li>接种物制备<ol><li>将质量控制微生物 大肠杆 菌和93种 沙门氏菌 菌株在37°C下在Mueller-Hinton琼脂（MHA）平板上检测AST16-18小时3。</li>
		<li>根据C...

<ol>
	<li>Ramanan, L., 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=Antimicrobial+resistance-the+need+for+global+solutions.">Antimicrobial resistance-the need for global solutions.</a> The Lancet Infectious Diseases. 13 (12), 1057-1098 (2013).</li><li>Xiaonan, Z., Qing, Z., Thomas, S. P., Yuqing, L., Martha, R. J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=inPhocus:+Perspectives+of+the+application+of+bacteriophages+in+poultry+and+aquaculture+industries+based+on+Varms+in+China.">inPhocus: Perspectives of the application of bacteriophages in poultry and aquaculture industries based on Varms in China.</a> PHAGE: Therapy, Applications, and Research. 2 (2), 69-74 (2021).</li><li>CLSI. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Performance Standards for Antimicrobial Disk Susceptibility Tests. 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S., L&#246;ffler, B., R&#246;del, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+antimicrobial+susceptibility+testing+in+blood+culture+diagnostics+performed+by+direct+inoculation+using+the+VITEK&#174;-2+and+BD+PhoenixTM+platforms.">Rapid antimicrobial susceptibility testing in blood culture diagnostics performed by direct inoculation using the VITEK&#174;-2 and BD PhoenixTM platforms.</a> European Journal of Clinical Microbiology &amp; Infectious Diseases. 38 (3), 471-478 (2019).</li><li>Dupuis, G. <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+the+Cobasbact+automated+antimicrobial+susceptibility+testing+system.">Evaluation of the Cobasbact automated antimicrobial susceptibility testing system.</a> European Journal of Clinical Microbiology &amp; Infectious Diseases. 4 (2), 119-122 (1985).</li><li>Liu, Y., 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+system+of+bacterial+antimicrobial+resistance+detection+and+its+operation+method.">A system of bacterial antimicrobial resistance detection and its operation method.</a> China Patent. , (2019).</li><li>Liu, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+high+throughput+test+plate+for+screening+bacteriophage+of+zoonotic+pathogens+and+its+application.">A high throughput test plate for screening bacteriophage of zoonotic pathogens and its application.</a> China Patent. , (2022).</li><li>Adams, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Bacteriophages. , (1959).</li><li>Nair, A., Ghugare, G. S., Khairnar, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+appraisal+of+bacteriophage+isolation+techniques+from+environment.">An appraisal of bacteriophage isolation techniques from environment.</a> Microbial Ecology. 83 (3), 519-535 (2022).</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=.">.</a> Shandong veterinary antibiotic resistance system. , (2023).</li><li>Ming, 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=Comparison+of+the+results+of+96-dot+agar+dilution+method+and+broth+microdilution+method.">Comparison of the results of 96-dot agar dilution method and broth microdilution method.</a> Chinese Journal of Antibiotics. 43 (6), 729-733 (2018).</li><li>Laxminarayan, R., Klugman, K. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Communicating+trends+in+resistance+using+a+drug+resistance+index.">Communicating trends in resistance using a drug resistance index.</a> BMJ Open. 1 (2), e000135 (2011).</li><li>Chen, Y., 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=Assessing+antibiotic+therapy+effectiveness+against+the+major+bacterial+pathogens+in+a+hospital+using+an+integrated+index.">Assessing antibiotic therapy effectiveness against the major bacterial pathogens in a hospital using an integrated index.</a> Future Microbiology. 12, 853-866 (2017).</li><li>Ciccolini, M., Spoorenberg, V., Geerlings, S. E., Prins, J. M., Grundmann, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Using+an+index-based+approach+to+assess+the+population-level+appropriateness+of+empirical+antibiotic+therapy.">Using an index-based approach to assess the population-level appropriateness of empirical antibiotic therapy.</a> Journal of Antimicrobial Chemotherapy. 70 (1), 286-293 (2015).</li><li>Yanbo, L., 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=Preliminary+application+of+inoculation+system+for+high-throughput+drug+susceptibility+test.">Preliminary application of inoculation system for high-throughput drug susceptibility test.</a> China Poultry. 42 (6), 52-57 (2020).</li></ol>

琼脂稀释法已经得到广泛应用。高通量AST系统的原理是琼脂稀释法。该协议中的关键步骤之一是一次准确高通量转移 96 个接种物，该转移连续多次进行。为了完成这一关键步骤，96点阵接种器的引脚均匀且非常光滑。每个引脚的自然沉积体积约为 2 μL，在琼脂培养基表面聚集成小液滴，迅速吸收到琼脂中，不会流动或飞溅以造成交叉污染。第二个关键步骤是对大型AST数据进行统计处理。在高通量?...

随着抗菌药物被广泛用于预防细菌性传染病，抗菌素耐药性（AMR）已成为全球公共卫生问题1。抗击抗微生物药物耐药性是目前监测流行病病原体抗微生物药物耐药性以及敏感抗菌剂和溶菌噬菌体的协同治疗的主要任务2。
体外 抗菌药物敏感性试验 （AST） 是监测治疗和检测 AMR 水平的主要方法。它是抗菌药理学的重要组成部分，也是临床用药的重要基础。美国临床和实验室标准协会（CLSI）和欧洲药敏试验委员会（EUCAST）制定和修订了AST的国际标准，并不断修改和补充AST方法和断点，以确定某种“生物体-抗菌剂”组合的MIC为敏感（S）、耐药（R）或中间（I）3，4.
从 1980 年代到 1990 年代，自动微量肉汤稀释仪器迅速发展并应用于临床实践，例如 Alfred 60AST、VITEK System、FISTM 和 Cobasbact5、6、7。然而，这些仪器价格昂贵，需要高成本的耗材，并且它们的检测范围是为临床患者用药设计的5,6,7。由于这些原因，它们不适合兽医临床检查和大量高耐药菌株的检测。本研究开发了一种智能高通量AST/噬菌体筛选系统，包括96点基质接种器（图1）、图像采集转换器（图2）和相应的软件8，通过琼脂稀释法对一批细菌菌株同时对多种抗菌剂进行AST。此外，该系统还用于检测和分析噬菌体对抗菌耐药菌的裂解模式9，并从噬菌体库中高效筛选裂解噬菌体。该系统被发现高效、经济且易于操作。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/64785/64785fig01.jpg"> 图 1：96 点阵接种器的结构图。 1：接种针板;2：移动运营商;3：种子块;4：孵育板;5：底座;6：操作手柄;7：限位销。<a href="https://www.jove.com/files/ftp_upload/64785/64785fig01large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/64785/64785fig02.jpg"> 图2：图像采集转换器的结构图。 1：外壳;2：显示屏;3：图像采集室;4：检测板底座;5：出入库检测板;6：控制板;7：图像采集转换装置;8：光源;9：图像扫描仪。<a href="https://www.jove.com/files/ftp_upload/64785/64785fig02large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>96 well&#160; culture plate</td>
			<td>Beijing lanjieke Technology Co., Ltd</td>
			<td>11510</td>
			<td></td>
		</tr>
		<tr>
			<td>96-dot matrix AST image acquisition system</td>
			<td>Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences</td>
			<td>In-house software copyright</td>
			<td></td>
		</tr>
		<tr>
			<td>96-dot matrix inoculator&#160;</td>
			<td>Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences</td>
			<td>N/A</td>
			<td>Patented product</td>
		</tr>
		<tr>
			<td>Agar</td>
			<td>Qingdao hi tech Industrial Park Haibo Biotechnology Co., Ltd</td>
			<td>HB8274-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Amikacin&#160;</td>
			<td>Shanghai McLean Biochemical Technology Co., Ltd</td>
			<td>A857053</td>
			<td></td>
		</tr>
		<tr>
			<td>Amoxicillin</td>
			<td>Shanghai McLean Biochemical Technology Co., Ltd</td>
			<td>A822839</td>
			<td></td>
		</tr>
		<tr>
			<td>Ampicillin</td>
			<td>Shanghai McLean Biochemical Technology Co., Ltd</td>
			<td>A830931</td>
			<td></td>
		</tr>
		<tr>
			<td>Analytical balance</td>
			<td>Sartorius</td>
			<td>BSA224S</td>
			<td></td>
		</tr>
		<tr>
			<td>Automated calculation software for Lar index of AMR</td>
			<td>Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences</td>
			<td>In-house software copyright</td>
			<td></td>
		</tr>
		<tr>
			<td>Bacteria Salmonella strains</td>
			<td>Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences</td>
			<td>N/A</td>
			<td>Animal origin</td>
		</tr>
		<tr>
			<td>Bacterial resistance Lar index certification management system V1.0</td>
			<td>Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences</td>
			<td>In-house software copyright</td>
			<td></td>
		</tr>
		<tr>
			<td>Ceftiofur</td>
			<td>Shanghai McLean Biochemical Technology Co., Ltd</td>
			<td>C873619</td>
			<td></td>
		</tr>
		<tr>
			<td>Ciprofloxacin</td>
			<td>Shanghai McLean Biochemical Technology Co., Ltd</td>
			<td>C824343</td>
			<td></td>
		</tr>
		<tr>
			<td>Clavulanic acid</td>
			<td>Shanghai McLean Biochemical Technology Co., Ltd</td>
			<td>C824181</td>
			<td></td>
		</tr>
		<tr>
			<td>Clean worktable</td>
			<td>Suzhou purification equipment Co., Ltd</td>
			<td>SW-CJ-2D</td>
			<td></td>
		</tr>
		<tr>
			<td>Colistin sulfate</td>
			<td>Shanghai McLean Biochemical Technology Co., Ltd</td>
			<td>C805491</td>
			<td></td>
		</tr>
		<tr>
			<td>Culture plate</td>
			<td>Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences</td>
			<td>N/A</td>
			<td>Patented product</td>
		</tr>
		<tr>
			<td>Doxycycline</td>
			<td>Shanghai McLean Biochemical Technology Co., Ltd</td>
			<td>D832390</td>
			<td></td>
		</tr>
		<tr>
			<td>Enrofloxacin</td>
			<td>Shanghai McLean Biochemical Technology Co., Ltd</td>
			<td>E809130</td>
			<td></td>
		</tr>
		<tr>
			<td>Filter 0.22 &#956;m</td>
			<td>Millipore</td>
			<td>SLGP033RB</td>
			<td></td>
		</tr>
		<tr>
			<td>Florfenicol</td>
			<td>Shanghai McLean Biochemical Technology Co., Ltd</td>
			<td>F809685</td>
			<td></td>
		</tr>
		<tr>
			<td>Gentamicin</td>
			<td>Shanghai McLean Biochemical Technology Co., Ltd</td>
			<td>G810322</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass bottle 50 mL</td>
			<td>Xuzhou Qianxing Glass Technology Co., Ltd</td>
			<td>QX-7</td>
			<td></td>
		</tr>
		<tr>
			<td>High-throughput resistance detection system V1.0</td>
			<td>Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences</td>
			<td>In-house software copyright</td>
			<td></td>
		</tr>
		<tr>
			<td>Image acquisition converter</td>
			<td>Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences</td>
			<td>N/A</td>
			<td>Patented product</td>
		</tr>
		<tr>
			<td>Meropenem</td>
			<td>Shanghai McLean Biochemical Technology Co., Ltd</td>
			<td>M861173</td>
			<td></td>
		</tr>
		<tr>
			<td>Mueller-Hinton agar</td>
			<td>Qingdao hi tech Industrial Park Haibo Biotechnology Co., Ltd</td>
			<td>HB6232</td>
			<td></td>
		</tr>
		<tr>
			<td>Petri dish 60 mm x 15 mm</td>
			<td>Qingdao Jindian biochemical equipment Co., Ltd</td>
			<td>16021-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Petri dish 90 mm x 15 mm</td>
			<td>Qingdao Jindian biochemical equipment Co., Ltd</td>
			<td>16001-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Salmonella phages</td>
			<td>Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Shaker incubator</td>
			<td>Shanghai Minquan Instrument Co., Ltd</td>
			<td>MQD-S2R</td>
			<td></td>
		</tr>
		<tr>
			<td>Turbidimeter</td>
			<td>Shanghai XingBai Biotechnology Co., Ltd</td>
			<td>F-TC2015</td>
			<td></td>
		</tr>
		<tr>
			<td>Varms base type library system V1.0</td>
			<td>Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences</td>
			<td>In-house software copyright</td>
			<td></td>
		</tr>
		<tr>
			<td>Vertical high-pressure steam sterilizer</td>
			<td>Shanghai Shen&#39;an medical instrument factory</td>
			<td>LDZX-75L</td>
			<td></td>
		</tr>
		<tr>
			<td>Veterinary pathogen resistance testing management system</td>
			<td>Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences</td>
			<td>In-house software copyright</td>
			<td></td>
		</tr>
		<tr>
			<td>Veterinary resistance cloud monitoring and phage control platform V1.0</td>
			<td>Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences</td>
			<td>In-house software copyright</td>
			<td></td>
		</tr>
	</tbody>
</table>

application of the intelligent high-throughput antimicrobial sensitivity testing/phage screening system and lar index of antimicrobial resistance

为了提高耐药菌药敏试验（AST）和噬菌体高通量筛选的效率，降低检测成本，根据AST标准和临床与实验室标准协会（CLSI）制定的耐药性断点（R），开发了一种智能高通量AST/噬菌体筛选系统，包括96点阵接种器、图像采集转换器和相应的软件。采用智能高通量AST/噬菌体筛选系统，对山东省家禽中分离的1500株 沙门氏 菌菌株的AST和最小抑菌浓度（MIC）分布（R/8-8R）进行统计。Lar指数，即“较少的抗生素，较少的耐药性和残留，直到很少的抗生素”，是通过计算每个MIC的加权平均值并除以R得到的。与使用耐药性流行率来表征高度耐药菌株的抗菌素耐药性 （AMR） 程度相比，这种方法提高了准确性。对于AMR高的 沙门氏 菌菌株，利用该系统从噬菌体文库中高效筛选裂解噬菌体，并计算分析裂解谱图。结果表明，智能高通量AST/噬菌体筛选系统具有可操作性、准确性、高效、成本低廉、易于维护等特点。该系统与山东省兽用抗菌素耐药性监测系统相结合，适用于与AMR相关的科学研究和临床检测。

遵循智能高通量AST系统的方案，以中国山东家禽 沙门氏菌 为例说明其应用。
沙 门氏菌 菌株在含有氨苄青霉素（R为32μg/ mL）的琼脂平板上生长，浓度为2至256μg/ mL，由图像采集转换器确定，如图 3所示。水平1号 井A1为阴性对照，未显示菌落生长;A2和A3为CLSI标准化的质控范围（2-8 μg/mL）为质控菌株，MIC为4 μg/mL（用2 μg/mL氨苄青霉素...

Watch this Scientific Journal Video about Application of the Intelligent High-Throughput Antimicrobial Sensitivity Testing/Phage Screening System and Lar Index of Antimicrobial Resistance at JoVE.com

Application of the Intelligent High-Throughput Antimicrobial Sensitivity Testing/Phage Screening System and Lar Index of Antimicrobial Resistance

在这里，我们介绍了智能高通量抗菌药敏检测/噬菌体筛选系统的原理、结构和说明。以中国山东省家禽中分离的 沙门氏菌 为例，说明了其应用。计算Lar指数，并全面讨论其在评估抗菌素耐药性方面的意义。

智能高通量抗菌药敏检测/噬菌体筛选系统及Lar耐药性指数的应用

To improve the efficiency of antimicrobial susceptibility testing (AST) and phage high-throughput screening for resistant bacteria and to reduce the ...

该协议提出了一种多层次的方法，用于EST检测和噬菌体图。它还引入了用于保存的大索引，以比较不同控制技术的 EMR 水平。这种具有成本效益的技术是智能、高通量和多功能的。它促成了VAMS的建立，这是一个兽医监测网络，收集了20， 000个流行基因和分析噬菌体。该方法还可用于医学、农业、动物心脏电池和渔业中的病原菌的监测和控制。该协议和本公司文件应提供。我们提供技术培训指导。遵循它将帮助任何人在短时间内掌握该技术。首先在Mueller-Hinton琼脂平板上孵育质量控制微生物大肠杆菌和93个测试沙门氏菌菌株。接下来，将每个菌株的制备接种物稀释10次。将 200 微升无菌生理盐水作为阴性对照转移到 96 孔板的 A-1 孔中。然后将两种大肠杆菌悬浮液分别移入a-2和a-3孔中，作为阳性和对照和质量控制。在剩余的 93 个孔中，加入 200 微升稀释的测试菌株。用于制备抗菌琼脂平板。首先，根据 LAR 指数设置被测不同抗菌剂的浓度范围。按照CLSI标准琼脂稀释方法对抗生素溶液进行log-two加倍稀释方案。现在将两毫升抗菌溶液加入含有 18 毫升熔融 Mueller-Hinton 琼脂培养基的灭菌玻璃瓶中。充分混合后，倒入生物安全柜内的板中。让琼脂在室温下凝固。确保在平板盖下留出间隙，以便琼脂表面干燥。在孵育板的底部标记抗菌剂的类型及其各自的浓度。将每种抗菌剂的多个孵育板排列在堆栈中，按对数二加倍稀释顺序排列。最后，准备两个无药物琼脂平板作为每种药物的对照。在生物安全柜内 96 点基质接种器的支架上安装灭菌接种针板。接下来，将准备好的种子块与测试菌株和琼脂孵育板放在移动载体上。推动移动载体，直到种子块位于接种针板下方。现在按下操作手柄并降低接种针板，将 96 针引导至种子块 96 孔中的接种物。以受控方式松开操作手柄，自动复位接种针板，充分搅拌各接种物并浸入，推动操作手柄两到三次。然后，移动并定位载板，使培养板位于接种针板下方。按下操作手柄，放下接种针板并放置一到两秒钟，以确保与培养板表面完全接触。松开操作手柄，完成一轮接种。更换孵育的平板并重复该过程，直到完成一批抗菌琼脂平板。切换到新的接种针板和种子块以接种另一组测试菌株。在室温下孵育接种的抗菌琼脂平板，直到接种斑点中的水分完全吸收到琼脂中。孵育后，倒置平板并继续孵育，使不受抑制的细菌形成菌落。要进行图像采集和数据统计，首先单击96点阵AST图像采集系统以启动程序。从任务栏中，选择“测试信息”。单击“新建”以创建新的测试任务并填写所需的详细信息。然后按数据收集，然后按照片和测试项目，选择新创建的任务。选择抗生素以选择抗生素名称，选择梯度以选择抗生素的初始浓度。最后，单击“连接”以与图像采集转换器建立连接。接下来，将孵育板放在检测板底座上，并将其插入图像采集转换器。单击集合以捕获板的图像。按抗生素并选择下一组板。选择渐变以识别起始渐变，然后继续进行下一轮图像采集。通过单击提交来完成该过程，以确定菌落形成并将图像转换为最小抑制浓度，即MIC值。单击查询以访问所有 MIC 结果。首先利用双层琼脂法生产不同的噬菌体。将噬菌体稀释至合适的平行浓度，并将200微升噬菌体接种物加入96孔种子块中。接下来，为要检查的每个菌株准备一个双层孵育板。在双层板的盖子下留出一个间隙，以便干燥。将制备好的噬菌体种子块和双层板放在96点阵接种器的移动载体上。将所有噬菌体接种物转移到半琼脂表面。让平板保持在室温下，直到所有接种水分都被吸收到半琼脂中。现在，在 37 摄氏度下为测试菌株孵育平板 4 到 6 小时。使用图像采集转换器获取并保存各双层板实验结果的图像。根据获得的图像，在电子表格中记录不同斑点形状的数量和形态，并计算不同类型噬菌体的各自比例。沙门氏菌在氨苄青霉素平板上生长的形态学研究表明，a-1中的阴性对照没有生长。a-4 中沙门氏菌菌株的 MIC 为 64 μg/mL，而 a-5 的 MIC 为 16 μg/mL。氨苄西林、环丙沙星和阿莫西林-克拉维酸的耐药百分比值高于50%，而多西环素、氟非尼考、头孢噻肟和恩诺沙星的耐药率值为30%-50%庆大霉素、阿米卡星和美罗培南的耐药率值小于30%，环丙沙星和阿莫西林-克拉维酸的LAR指数明显延迟。沙门氏菌在双层板上生长的形态学分析表明，观察到噬菌体的4个主要斑点形态。圆形透明裂解点来自噬菌体，该噬菌体可以可靠地杀死平板上的宿主，但可能会也可能不会成功复制，代价是该宿主。牌匾的集合是由真正的牌匾形成的。浑浊的裂解点是由噬菌体引起的，该噬菌体不能可靠地杀死平板上的宿主，并且可能会也可能不会成功复制，而代价是该宿主。无裂解点表示非裂解性质。该技术可轻松有效地一次转移 96 个琼脂透镜，仪器会自动识别图像，然后计算线骰子。该技术有望促进噬菌体的研究和产业化，噬菌体的应用是减少和解决AMR问题的惊人方法之一。

Research

JoVE Journal

Environment

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

在使用高通量微孔板测定水，土壤，沉积物和微生物测定胞外酶活性的

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

科研

环境科学

High-throughput Fluorometric Measurement of Potential Soil Extracellular Enzyme Activities

潜在土壤胞外酶活性的高通量荧光测量

Microbes in soils and other environments produce extracellular enzymes to depolymerize and hydrolyze organic macromolecules so that they can be ...

Optimized Negative Staining: a High-throughput Protocol for Examining Small and Asymmetric Protein Structure by Electron Microscopy

优化负染：通过电子显微镜检查小和不对称蛋白结构的高通量协议

Structural determination of proteins is rather challenging for proteins with molecular masses between 40 - 200 kDa. Considering that more than half of ...

Application of Two-spotted Spider Mite Tetranychus urticae for Plant-pest Interaction Studies

Application of Two-spotted Spider Mite Tetranychus urticae for Plant-pest Interaction Studies

二斑叶螨的应用<em&gt;二斑叶螨</em&gt;植物病虫害的相互作用研究

The two-spotted spider mite, Tetranychus urticae, is a ubiquitous polyphagous arthropod herbivore that feeds on a remarkably broad array of species, with ...

Modifying the Bank Erosion Hazard Index (BEHI) Protocol for Rapid Assessment of Streambank Erosion in Northeastern Ohio

Modifying the Bank Erosion Hazard Index BEHI Protocol for Rapid Assessment of Streambank Erosion in Northeastern Ohio

修改银行侵蚀危险指数（BEHI）协议的河岸侵蚀美国俄亥俄州东北部的快速评估

Understanding the source of pollution in a stream is vital to preserving, restoring, and maintaining the stream&#8217;s function and habitat it provides. ...

Semi-High Throughput Screening for Potential Drought-tolerance in Lettuce (Lactuca sativa) Germplasm Collections

Semi-High Throughput Screening for Potential Drought-tolerance in Lettuce Lactuca sativa Germplasm Collections

半高通量筛选潜在的抗干旱的生菜（<em&gt;莴苣</em&gt;）种质资源

This protocol describes a method by which a large collection of the leafy green vegetable lettuce (Lactuca sativa L.) germplasm was screened for likely ...

High-throughput Screening of Recalcitrance Variations in Lignocellulosic Biomass: Total Lignin, Lignin Monomers, and Enzymatic Sugar Release

总木质素，木质素单体，和酶促糖推出：顽拗变奏曲在木质纤维素生物质的高通量筛选

The conversion of lignocellulosic biomass to fuels, chemicals, and other commodities has been explored as one possible pathway toward reductions in the ...

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

百里香精油应用于肉类干燥过程中微生物负荷的影响

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 ...

The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea

The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea

植物感染试验: 以植物病原菌为载体的喷雾和伤口介导接种

Plants possess a powerful system to defend themselves against potential threats by pathogenic fungi. For agriculturally important plants, however, current ...

High-throughput, Microscale Protocol for the Analysis of Processing Parameters and Nutritional Qualities in Maize (Zea mays L.)

High-throughput, Microscale Protocol for the Analysis of Processing Parameters and Nutritional Qualities in Maize Zea mays L.

玉米加工参数和营养品质分析的高通量、微尺度协议 (玉蜀黍)

Maize is an important grain crop in the United States and worldwide. However, maize grain must be processed prior to human consumption. Furthermore, whole ...