啤酒酿造过程中有害微生物的危害及其检测

本文分析了有害微生物对啤酒质量的影响及其有害微生物的检测方法，着重介绍了快速生物检测方法——PCR技术在啤酒污染微生物检测中的应用。     

啤酒有害菌检测及NBB的使用

啤酒在酿造、包装和储存过程中，真正对啤酒产生危害的是那些能在啤酒中生长，引起酒体浑浊，产生异味的厌氧茵和微好氧茵。而NBB培养基是一种检测啤酒有害茵选择性较好的培养基，它快速、可靠、选择性高。啤酒企业开展啤酒有害茵的检验和控制，对提高啤酒质量及其生物稳定性具有十分重要的意义。     

啤酒腐败细菌与酒花抗性

啤酒的腐败细菌有革兰氏阳性菌（乳酸菌、片球菌、M．kristinae等）和革兰氏阴性菌（Pectinatus、巨型球菌、Z．raffinosivorans等）。啤酒中的酒花具有抑制腐败菌的作用，主要物质为异α-酸；酒花能对微生物体内的酶起药物钝化、改变目的底物、抑制药物流入等作用。对啤酒腐败菌的检测方法有聚合酶链式反应（PCR）、伏安型生物传感器检测鉴定微生物细胞、自动微生物检测系统（AMS）、改良MRS培养基检测啤酒中乳酸茵和ATP法。（孙悟）     

PCR技术在食品微生物检测中的运用

由于受到环境影响,部分原生食物中污染成分含量较高,致使其出现了食品安全隐患。部分食物中的有害菌群远高出限定值,或存在致病微生物,为此需要利用微生物检测方法对食品进行检测。本文首先阐述了PCR技术的应用原理,然后分析了PCR技术对致病菌、乳酸菌、啤酒腐败菌的检测。     

foodproof(R)啤酒检测试剂盒一次性检测30种啤酒有害菌

在啤酒酿造过程中,对快速检测有害茵有非常高的要求.过去,日常应用的方法有很多,它们大多数不是工序过于繁琐,就是检测不具有专一性;应用传统检测方法至少需要6天甚至更长时间才能获得检测结果.传统的检测方法越来越不能满足现代高速啤酒生产线需求;啤酒生产企业的质量管理人员亟需一种能快速检测啤酒有害菌的方法.茵群的种类有快速生长型和缓慢生长型.如果了解所污染微生物的种类,那么就能够就污染源进行有效跟踪,当污染种群发生变化也能及时发现.所有这些都能帮助企业降低生产损失和减少额外清洗工作,还可以避免产品召回,提高啤酒企业的生产效率.     

控制CIP系统啤酒有害菌污染的重要性

众所周知,在啤酒生产中,除了人工纯种培养的酵母菌之外,不允许其他微生物侵入,尤其是啤酒的有害菌。如何解决有害菌的污染,生产出高品质啤酒,是啤酒厂质量控制的重要内容。国内啤酒厂家微生物检验工作起步较晚,检测点少,检测方法不规范,微生物检测点重点放在啤酒     

浅谈纯生啤酒生产中微生物控制

纯生啤酒采用无菌酿造、无菌过滤、无菌包装,生产纯生啤酒的关键是防止和控制有害微生物的污染,即从生产的各个环节入手对微生物进行严格管理和控制,以保证啤酒的微生物安全,它包括纯种酿造、无菌过滤、无茵罐装等过程.     

啤酒酿造过程中的微生物控制

微生物控制是啤酒酿造过程中的重要环节。介绍了啤酒酵母的使用，一般微生物控制方法及有害菌的检测。     

实时荧光PCR方法快速检测鉴定啤酒有害菌的研究

本文对PCR方法快速检测鉴定啤酒有害菌进行了研究,结果表明使用实时荧光PCR方法检测啤酒有害菌可以将检测时间缩短至48h,并可以同时得到检测与鉴定的结果。该方法有较好的特异性、可重复性,相比传统培养基法,PCR法的检测结果准确可靠。     

啤酒酿造过程中有害微生物的防治

啤酒酿造是利用啤酒酵母进行的纯种发酵。本文首先分析了啤酒生产中常见的有害微生物及对啤酒质量的危害，然后针对有害微生物的来源，提出了相应的防治措施。     

啤酒有害菌的PCR检测和SYBR Green实时PCR定量

啤酒有害菌是一些能在啤酒中存活并使啤酒的外观和风味发生改变的细菌,对其进行快速检测和定量是啤酒生产急待解决的问题。我们从华润雪花啤酒（中国）有限公司各工厂提供的样品中分离到28株啤酒有害菌,16S rDNA序列的系统进化分析表明,其中26个菌株属于乳杆菌属（Lactobacillus spp．）、1个菌株为明串珠菌属（Leuconostoc spp．）,1个菌株为片球菌属（Pediococcu sp．）。根据酒花（hop）抗性基因horA、horB和horC的保守序列设计了扩增这3个基因的PCR引物,用这些引物对28株啤酒有害菌进行了常规PCR检测,检出率分别为89％、79％和75％,用hor A—horC双引物进行检测,检出率为100％。用SYBR Green实时定量PCR技术,以horA基因为靶序列,建立了对啤酒有害菌的细胞数进行快速定量的新方法,用该方法测定的污染啤酒样品中有害菌的浓度与平板培养法相近。     

An improved plate culture procedure for the rapid detection of beer‐spoilage lactic acid bacteria

Lactic acid bacteria (LAB) are the most frequently encountered beer‐spoilage bacteria, and they can render beer undrinkable owing to the production of lactic acid, diacetyl and turbidity. Three beer‐spoilage strains, 2011–6, 2011–8 and 2011–11, were isolated from finished beers. Based on the 16S rRNA sequence analysis, these three isolates were identified as Lactobacillus acetotolerans. Only the horA homologue was detected in these strains, while the horC homologue was not detected. In addition, an improved plate culture method for the rapid detection of beer‐spoilage LAB by the addition of catalase was evaluated. Supplementation with catalase enhanced the growth and colony sizes of the spoilage LAB investigated. These beer‐spoilage bacteria, including some slowly growing strains, were detected within five days of incubation using the modified method. Taken together, the modified procedure could be a rapid countermeasure against beer‐spoilage LAB, and it compared favourably with the conventional plate culture method. Copyright © 2014 The Institute of Brewing & Distilling     

EMA-PCR法快速检测啤酒中腐败短乳杆菌

将叠氮溴乙锭(ethidium bromide monoazide,EMA)与聚合酶链式反应(polymerase chain reaction,PCR)技术相结合,以酒花耐受基因hor C为靶基因,用啤酒腐败短乳杆菌基因组DNA作为模板进行扩增。结果发现,在前处理过程中加入EMA,当终质量浓度小于20μg/m L时,对活的短乳杆菌中靶基因的扩增没有明显抑制作用;而当EMA终质量浓度为1.0μg/m L时可有效抑制10~5 CFU/m L短乳杆菌死细胞的扩增。本实验建立的EMA-PCR检测方法的灵敏度为10~4活细胞/m L酒液样品。验证实验结果表明,在13株乳酸菌中,建立的hor C特异性EMA-PCR能有效检测到其中的全部5株啤酒污染菌,同时可区分这5株菌的活/死细胞混合体系,降低检测过程中的假阳性。     

聚合酶链式反应(PCR)技术鉴定啤酒腐败菌的最新进展

综述了PCR技术在啤酒腐败细菌鉴定方面的应用及最新进展 ;介绍了 3种PCR技术 ,特别讲述了利用这些技术对啤酒中乳酸菌的鉴定 ;并分析了PCR技术的局限性 ,对其应用前景进行了展望。     

啤酒花及其抗菌机理研究进展

就国内外对酒花的研究、啤酒花的发展及其应用历史、啤酒花的成分、酒花抗菌物质进行了分析和综述.重点对啤酒花的抗菌机理和抗菌特性进行了阐述.通过介绍一些常用和最新检测啤酒腐败菌的方法,论述了酒花抗菌机理对啤酒腐败菌检测的影响意义,展望了啤酒腐败菌检测的发展趋势.     

125th Anniversary Review: Microbiological Instability of Beer Caused by Spoilage Bacteria

Beer has been generally recognized as a microbiologically stable beverage. However, microbiological incidents occasionally occur in the brewing industry. The microbiological instability of beer is often caused by bacteria consisting of four genera, Lactobacillus, Pediococcus, Pectinatus and Megasphaera. Lactobacillus and Pediococcus belong to the lactic acid bacteria (LAB), whereas Pectinatus and Megasphaera form a group of strict anaerobes that are known as intermediates between Gram‐positive and Gram‐negative bacteria. The frequencies of beer spoilage incidents caused by these four genera have been reported to exceed 90% in Europe and therefore Lactobacillus, Pediococcus, Pectinatus and Megasphaera are considered to be the principal spoilage agents in the brewing industry. Thus, this review consists of three parts involving these four genera. The first part describes spoilage LAB in alcoholic beverages with some emphasis on beer spoilage LAB. In this part, the emergence and evolution of these spoilage LAB is discussed, the insight of which is useful for developing quality control methods for these beverages. The second part is devoted to the hop resistance in beer spoilage LAB. This area of research is evolving rapidly and recent progress in this field is summarized. The third part concerns Pectinatus and Megasphaera. Although this group of beer spoilage bacteria has been described relatively recently, the incident reports in Europe increased in the early 1990s, reaching around 30% of spoilage incidents. Various aspects of Pectinatus and Megasphaera, ranging from their taxonomy and beer spoilage ability to detection and eradication methods are described.     

Detection of culturable and viable but non‐culturable cells of beer spoilage lactic acid bacteria by combined use of propidium monoazide and horA‐specific polymerase chain reaction

Current methods of detecting beer spoilage lactic acid bacteria (LAB) are time‐consuming and do not differentiate between viable and non‐viable bacteria. In this study, a combination of the conventional polymerase chain reaction (PCR) and propidium monoazide (PMA) pretreatment has been described to circumvent the disadvantages. The horA‐specific PMA‐PCR described here identifies beer spoilage LAB based not on their identity, but on the presence of a gene that is shown to be highly correlated with the ability of LAB to grow in beer. The results suggest that the use of 20 µg/mL or less of PMA did not inhibit the PCR amplification of DNA derived from viable, but putatively non‐culturable (VPNC) Lactobacillus acetotolerans. The minimum amount of PMA to completely inhibit the PCR amplification of DNA derived from dead L. acetotolerans cells was 1.5 µg/mL. The detection limit of established PMA‐PCR assays was found to be 100 VPNC cells/reaction for the horA gene. Furthermore, the horA‐specific PMA‐PCR assays were subjected to 18 reference strains, representing 100% specificity with no false positive amplification observed. In conclusion, the use of horA‐specific PMA‐PCR allows for a substantial reduction in the time required for the detection of potential beer spoilage LAB and efficiently discriminates between live and dead cells. Copyright © 2016 The Institute of Brewing & Distilling     

PCR技术检测啤酒中腐败菌的研究进展

对传统方法和PCR技术检测啤酒中腐败菌的原理进行了分析和比较，对传统检测方法更有优势的PCR检测技术进行了综述。综合分析了目前国内外PCR技术检测的最新研究，重点讨论了标志性基因（抗酒花基因）和引物设计在PCR检测技术中应用，并对PCR技术检测啤酒中腐败菌的应用前景进行了分析。     

啤酒污染乳酸菌PCR引物的设计与验证

根据细菌16SrDNA序列的特点,对啤酒中常见污染乳酸菌16SrDNA序列进行分析,设计合成了针对啤酒污染乳酸菌的特征引物。并用该引物对从啤酒厂分离到的7种乳酸菌进行了检测,PCR结果表明该引物能够准确检测到啤酒中常见的乳酸菌。     

细菌污染对啤酒的影响

概述了重要细菌群的特征 ,及其初始和二级代谢终产物对啤酒香味风味的影响 ,提出了针对主要污染菌在生产过程中应采取的有效微生物控制和卫生措施