Guojie Cao, Jennifer Miller, Shizhen Steven Wang, Sandra Tallent
{"title":"BACARA®2与RAPID’b的计数比较研究。蜡样芽孢杆菌®琼脂对甘露醇-蛋黄-多粘菌素（MYP）琼脂的检测","authors":"Guojie Cao, Jennifer Miller, Shizhen Steven Wang, Sandra Tallent","doi":"10.1093/jaoacint/qsaf059","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>The Bacillus cereus group (B. cereus sensu lato) is a group of spore-forming strains. B. cereus group is a human pathogen associated with foodborne outbreaks with symptoms of diarrhea and emesis.</p><p><strong>Objective: </strong>This study evaluates the performance of BACARA® 2 (BAC2) and RAPID'B. cereus® (RAPID) agars compared to mannitol-egg yolk-polymyxin (MYP) agars and the original BACARA® (BAC) media to detect and enumerate the B. cereus group in pure culture and foods.</p><p><strong>Methods: </strong>We incorporated five food types to include liquid milk, whey powder, mashed potatoes, rice, and tea bags. For each food matrix, there were five test portions per level inoculum prepared (low, medium, and high), and a negative control. Individual test portions were diluted 1:10 in Butterfield's phosphate buffer (BPB) and 0.1 mL of the dilutions were inoculated on BAC2, RAPID, and MYP agars. In-house R script was applied to conduct statistical analysis to compare medium performance in food adulteration tests.</p><p><strong>Results: </strong>Inclusivity testing determined that all six B. cytotoxicus strains grew on BAC2, RAPID, and MYP but only one strain grew on BAC after 24 h incubation. RAPID had similar results as BAC2 in single-colony tests. In food tests, relative level of detection (RLOD) values indicated MYP was more sensitive for detecting B. cereus strains than BAC2 and RAPID, which is a result of B. cytotoxicus strains in mashed potato and whey powder needing an additional 24-hour incubation to exhibit typical morphology on BAC2 and RAPID.</p><p><strong>Conclusion: </strong>Our study demonstrated the use of BAC2 and/or RAPID agar will improve the performance of the U.S. Food and Drug Administration Bacteriological Analytical Manual (BAM) B. cereus method, particularly for the detection of B. cytotoxicus. Importantly, the use of BAC2 and/or RAPID will decrease the time to detect viable B. cereus strains without additional confirmation steps compared to MYP.</p><p><strong>Highlights: </strong>For improved isolation and identification of B. cereus group in foods, we propose the use of BAC2 or RAPID as optional agars, along with MYP.</p>","PeriodicalId":94064,"journal":{"name":"Journal of AOAC International","volume":" ","pages":"732-752"},"PeriodicalIF":1.7000,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12418170/pdf/","citationCount":"0","resultStr":"{\"title\":\"Enumeration Comparison Study of BACARA® 2 and RAPID'B. cereus® Agars to Mannitol-Egg Yolk-Polymyxin (MYP) Agar for Detection of Bacillus cereus Group Strains.\",\"authors\":\"Guojie Cao, Jennifer Miller, Shizhen Steven Wang, Sandra Tallent\",\"doi\":\"10.1093/jaoacint/qsaf059\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>The Bacillus cereus group (B. cereus sensu lato) is a group of spore-forming strains. B. cereus group is a human pathogen associated with foodborne outbreaks with symptoms of diarrhea and emesis.</p><p><strong>Objective: </strong>This study evaluates the performance of BACARA® 2 (BAC2) and RAPID'B. cereus® (RAPID) agars compared to mannitol-egg yolk-polymyxin (MYP) agars and the original BACARA® (BAC) media to detect and enumerate the B. cereus group in pure culture and foods.</p><p><strong>Methods: </strong>We incorporated five food types to include liquid milk, whey powder, mashed potatoes, rice, and tea bags. For each food matrix, there were five test portions per level inoculum prepared (low, medium, and high), and a negative control. Individual test portions were diluted 1:10 in Butterfield's phosphate buffer (BPB) and 0.1 mL of the dilutions were inoculated on BAC2, RAPID, and MYP agars. In-house R script was applied to conduct statistical analysis to compare medium performance in food adulteration tests.</p><p><strong>Results: </strong>Inclusivity testing determined that all six B. cytotoxicus strains grew on BAC2, RAPID, and MYP but only one strain grew on BAC after 24 h incubation. RAPID had similar results as BAC2 in single-colony tests. In food tests, relative level of detection (RLOD) values indicated MYP was more sensitive for detecting B. cereus strains than BAC2 and RAPID, which is a result of B. cytotoxicus strains in mashed potato and whey powder needing an additional 24-hour incubation to exhibit typical morphology on BAC2 and RAPID.</p><p><strong>Conclusion: </strong>Our study demonstrated the use of BAC2 and/or RAPID agar will improve the performance of the U.S. Food and Drug Administration Bacteriological Analytical Manual (BAM) B. cereus method, particularly for the detection of B. cytotoxicus. Importantly, the use of BAC2 and/or RAPID will decrease the time to detect viable B. cereus strains without additional confirmation steps compared to MYP.</p><p><strong>Highlights: </strong>For improved isolation and identification of B. cereus group in foods, we propose the use of BAC2 or RAPID as optional agars, along with MYP.</p>\",\"PeriodicalId\":94064,\"journal\":{\"name\":\"Journal of AOAC International\",\"volume\":\" \",\"pages\":\"732-752\"},\"PeriodicalIF\":1.7000,\"publicationDate\":\"2025-09-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12418170/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of AOAC International\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1093/jaoacint/qsaf059\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of AOAC International","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1093/jaoacint/qsaf059","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
背景：蜡样芽孢杆菌群（B. cereus sensu lato）是一组孢子形成菌株。灰状芽孢杆菌群是一种与腹泻和呕吐症状的食源性暴发有关的人类病原体。目的：本研究评价BACARA®2 （BAC2）和RAPID’B. cereus®（RAPID）琼脂与甘露醇-蛋黄-多粘菌素（MYP）琼脂和原BACARA®（BAC）培养基在纯培养物和食品中检测和枚举蜡样芽孢杆菌群的性能。方法：选取液态奶、乳清粉、土豆泥、大米、茶包等5种食物。对于每种食物基质，每个接种水平准备了5个试验部分（低、中、高），并进行了阴性对照。将单个测试部分用1:10的BPB稀释，并将0.1 mL的稀释液接种于BAC2、RAPID和MYP琼脂上。采用内部R脚本进行统计分析，比较食品掺假检测的中效。结果：包容性测试表明，所有6株细胞毒杆菌菌株在BAC2、RAPID和MYP上都能生长，但在BAC上孵育24小时后，只有1株菌株能生长。在单菌落试验中，RAPID与BAC2的结果相似。在食品试验中，RLOD值表明MYP对蜡样芽孢杆菌的检测比BAC2和RAPID更敏感，这是由于土豆泥和乳清粉中的细胞毒性芽孢杆菌菌株需要额外孵育24小时才能在BAC2和RAPID上呈现典型形态。结论：我们的研究表明，使用BAC2和/或RAPID琼脂可以提高BAM蜡样芽孢杆菌法的性能，特别是对细胞毒性芽孢杆菌的检测。重要的是，与MYP相比，使用BAC2和/或RAPID将减少检测活蜡样芽孢杆菌菌株的时间，而无需额外的确认步骤。为了更好地分离和鉴定食品中的蜡样芽孢杆菌群，我们建议使用BAC2或RAPID作为可选琼脂，以及MYP。
Enumeration Comparison Study of BACARA® 2 and RAPID'B. cereus® Agars to Mannitol-Egg Yolk-Polymyxin (MYP) Agar for Detection of Bacillus cereus Group Strains.
Background: The Bacillus cereus group (B. cereus sensu lato) is a group of spore-forming strains. B. cereus group is a human pathogen associated with foodborne outbreaks with symptoms of diarrhea and emesis.
Objective: This study evaluates the performance of BACARA® 2 (BAC2) and RAPID'B. cereus® (RAPID) agars compared to mannitol-egg yolk-polymyxin (MYP) agars and the original BACARA® (BAC) media to detect and enumerate the B. cereus group in pure culture and foods.
Methods: We incorporated five food types to include liquid milk, whey powder, mashed potatoes, rice, and tea bags. For each food matrix, there were five test portions per level inoculum prepared (low, medium, and high), and a negative control. Individual test portions were diluted 1:10 in Butterfield's phosphate buffer (BPB) and 0.1 mL of the dilutions were inoculated on BAC2, RAPID, and MYP agars. In-house R script was applied to conduct statistical analysis to compare medium performance in food adulteration tests.
Results: Inclusivity testing determined that all six B. cytotoxicus strains grew on BAC2, RAPID, and MYP but only one strain grew on BAC after 24 h incubation. RAPID had similar results as BAC2 in single-colony tests. In food tests, relative level of detection (RLOD) values indicated MYP was more sensitive for detecting B. cereus strains than BAC2 and RAPID, which is a result of B. cytotoxicus strains in mashed potato and whey powder needing an additional 24-hour incubation to exhibit typical morphology on BAC2 and RAPID.
Conclusion: Our study demonstrated the use of BAC2 and/or RAPID agar will improve the performance of the U.S. Food and Drug Administration Bacteriological Analytical Manual (BAM) B. cereus method, particularly for the detection of B. cytotoxicus. Importantly, the use of BAC2 and/or RAPID will decrease the time to detect viable B. cereus strains without additional confirmation steps compared to MYP.
Highlights: For improved isolation and identification of B. cereus group in foods, we propose the use of BAC2 or RAPID as optional agars, along with MYP.
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
