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1.
杨波  潘丽晶 《中国酿造》2006,(10):42-45
根据乳酸菌16SrDNA序列的特征,设计合成了针对啤酒有害乳酸菌的通用引物,在16SrDNA基因水平上采用聚合酶链式反应(PCR)技术鉴定了啤酒中59株有害乳酸菌的种类。同时根据鉴定结果设计了8种乳酸菌的特异引物,PCR技术验证结果表明该8种特异引物能够准确鉴定乳酸菌。  相似文献   

2.
啤酒污染乳酸菌PCR引物的设计   总被引:6,自引:0,他引:6  
郑飞云  金建中  顾国贤 《酿酒》2002,29(2):44-47
根据细菌16s rDNA序列的特点,通过对啤酒污染菌16s rDNA序列进行分析,设计合成了针对啤酒污染乳酸菌的引物,在16srDNA基因水平证明了该引物对乳酸菌的特异性,该引物的特异性是PCR检测技术在啤酒厂推广应用的前提,同时反映了16s rDNA在微生物鉴定中所起的重要作用。  相似文献   

3.
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。  相似文献   

4.
基于16s rDNA的PCR快速鉴定啤酒腐败菌的研究   总被引:5,自引:0,他引:5  
田小群  周世宁 《酿酒》2006,33(2):51-54
从Genebank中下载啤酒腐败菌所在属的63种细菌的16s rDNA序列,用DNAstar软件进行序列对比分析,找出这63种细菌16s rDNA序列所共有的5条同源序列,以这5条同源序列为基础设计出一对适合所有腐败菌的通用引物:上游:5’-GAACAGGATTAGATACCC-3’,下游:5’-GACTFAACCCAACATCTCAC-3’。利用该对引物,可以检测出啤酒厂出现的8种腐败菌,而对啤酒厂出现的13种非腐败菌则不能检出,保证了该引物对啤酒腐败菌的特异性。  相似文献   

5.
本研究主要评价RAPD-PCR技术在快速鉴定啤酒污染菌中的应用。首先评价了CTAB法和试剂盒提取DNA模板对RAPD指纹稳定性的影响以及乳酸菌专一性BP引物扩增的16SrDNA的5'末端740bp序列用于鉴定菌种的可行性,采用PCR产物直接测序鉴定分离的污染菌,构建标准污染菌库。对分离菌M-13的RAPD指纹聚类分析表明,相同来源的同一种菌能很好地归类在一起。根据建立的快速鉴定流程和初步确定的菌种相似性阈值(SCC),对8株分离菌的鉴定表明,RAPD-PCR技术是一项简单、快捷、可靠性强的鉴定技术,为研究啤酒酿造过程的污染菌提供了一个非常有用的快速鉴定工具。  相似文献   

6.
啤酒腐败菌的分离鉴定及对啤酒的危害   总被引:1,自引:0,他引:1  
从啤酒厂分离到31株啤酒腐败菌。对其进行了AP150CHL试剂条生理生化反应试验。并测定了这3l株菌的16SrDNA部分序列,与Genebank中已知序列进行了比较,并以此为依据进行了系统进化关系分析。结果表明,31株啤酒腐败菌分属于5个种,部分腐败菌对啤酒风味有不良影响。  相似文献   

7.
为了研究内蒙古锡林郭勒盟传统酸马奶的乳酸菌构成,将从酸马奶样品中分离到的11株乳酸菌采用16 SrDNA鉴定方法进行分子鉴定。结果为:将所测菌株序列与标准菌株序列进行比对,菌株序列发现与瑞士乳杆菌菌株序列的同源性均较高,所测11株乳酸菌菌株均为瑞士乳杆菌。  相似文献   

8.
乳酸菌对制麦和啤酒酿造的影响   总被引:1,自引:0,他引:1  
利用乳酸菌生物酸化麦芽、糖化醪和麦芽汁酿造的啤酒,有利于啤酒生物稳定性。有些乳酸菌产生乳酸菌素,能抑制某些革兰氏阳性腐败菌的生长。乳酸菌也是霉菌的天然抗菌剂,降低啤酒中霉菌毒素的形成。但大多数乳酸菌是啤酒主要的污染菌,对啤酒质量影响很大。腐败乳酸菌产生生物胺,可以用来检测污染程度。  相似文献   

9.
奶粉中阪崎肠杆菌PCR和荧光PCR 检测方法的研究   总被引:3,自引:0,他引:3  
高虹  张霞  高旗利 《食品科学》2006,27(9):203-207
建立了奶粉中可致婴幼儿高死亡率的阪崎肠杆菌的PCR和荧光PCR检测方法。利用细菌16S和23SrDNA的保守区设计通用引物,对6株阪崎肠杆菌16S-23SrDNA间区序列(ITS)进行扩增和测序,在比对阪崎肠杆菌ITS序列的基础上,设计了11条PCR和荧光PCR检测引物,组合成30对PCR引物,并筛选出一对种特异性引物,建立了奶粉中阪崎肠杆菌PCR和荧光PCR检测方法。用10株阪崎肠杆菌,18株近源菌株验证实验表明,本文所建立的PCR和荧光PCR方法特异性强;加菌实验表明,奶粉样品中阪崎肠杆菌检测低限为(2.2~5.4)CFU/100g,灵敏度高;新建的PCR和荧光PCR方法与FDABAM(美国食品及药品管理局微生物分析手册)方法比对实验表明,三种方法的检测结果完全一致。由于PCR和荧光PCR检测方法快速、可靠,因此可替代传统检验方法。  相似文献   

10.
结合传统的细菌分离培养法与现代分子生物技术方法16SrDNA菌群分析法对冰鲜鸭中优势腐败菌进行鉴定。传统分离培养检测到了17株优势腐败菌,16SrDNA菌群分析表明,冰鲜鸭中假单胞菌占绝对优势,达到54%;气单胞茵、肠杆菌是仅次于假单胞菌的第二个类群,都分别占15%;乳酸菌、节杆菌、紫色杆菌以及一株未鉴定的属都分别占4%。  相似文献   

11.
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  相似文献   

12.
The microbial flora of fresh meat stored aerobically at 5 degrees C up to spoilage was enumerated and collected in order to have mixed spoilage bacterial groups to be used in competition tests against Brochothrix thermosphacta. The bacterial groups collected as bulk colonies were identified by PCR-DGGE followed by partial 16S rDNA sequencing. The predominant bacteria associated with the spoilage of the refrigerated beef were B. thermosphacta, Pseudomonas spp, Enterobacteriaceae and lactic acid bacteria (LAB). The interactions between B. thermosphacta and the other spoilage microbial groups were studied in vitro at 5 degrees C. The results showed that a decrease of the growth of B. thermosphacta was evidenced in presence of LAB at 5 degrees C while the bacterium is the dominant organism when inoculated with mixtures of Pseudomonas spp., LAB and Enterobacteriaceae. A better understanding of bacterial meat spoilage interactions may lead to improved quality of fresh meat stored in refrigerated conditions.  相似文献   

13.
The presence of lactic acid bacteria (LAB) during shochu fermentation was monitored by PCR-denaturing gradient gel electrophoresis (DGGE) and by bacteriological culturing. No LAB were detected from fermented mashes by PCR-DGGE using a universal bacterial PCR primer set. However, PCR-DGGE using a new primer specific for the 16S rDNA of Lactococcus, Streptococcus, Tetragenococcus, Enterococcus, and Vagococcus and two primers specific for the 16S rDNA of Lactobacillus, Pediococcus, Leuconostoc, and Weissella revealed that Enterococcus faecium, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus nagelii, Lactobacillus plantarum, Lactococcus lactis, Leuconostoc citreum, Leuconostoc mesenteroides, and Weissella cibaria inhabited in shochu mashes. It was also found that the LAB community composition during shochu fermentation changed after the main ingredient and water were added during the fermentation process. Therefore, we confirmed that PCR-DGGE using all three primers specific for groups of LAB together was well suited to the study of the LAB diversity in shochu mashes. The results of DGGE profiles were similar to the results of bacteriological culturing. In conclusion, LAB are present during shochu fermentation but not dominant.  相似文献   

14.
Lactic acid bacteria are the most frequently encountered beer‐spoilage bacteria, and they may render beer undrinkable due to the production of lactic acid, diacetyl, and turbidity. Microbrewed beer is typically sold unpasteurised, leaving it more susceptible to spoilage by lactic acid bacteria. In this study, the incidence of lactic acid bacteria in bottled microbrewed beer from Victoria, Australia was investigated. A total of 80 beers from 19 breweries were screened for lactic acid bacteria. Almost 30% contained culturable lactic acid bacteria, and many had lactic acid levels well above the flavour threshold. Ethanol, hops, and the pH levels of the beers were not predictors for spoilage in the beers examined, and contamination appeared to be more closely linked to the source brewery. The 45 lactic acid strains isolated from these beers were identified by RAPD‐PCR, with Lactobacillus brevis being the most frequently isolated species. All isolates were capable of spoiling beer and contained putative hop resistance genes. At typical beer levels, pH and ethanol had no effect on the growth of the particular spoilage bacteria isolated in this study.  相似文献   

15.
为准确鉴定分离自我国四川鲜牦牛奶和曲拉中9株乳酸菌到种水平,作者运用16S rDNA序列分析、recA基因特异扩增和hsp60-RFLP等多种分子技术对分离自我国四川地区鲜牦牛奶和曲拉中的9株乳酸菌进行分类鉴定。结果证实,16S rDNA序列分析法可将9株乳酸菌初步归类为植物乳杆菌群(4株),肠膜明串珠菌(4株),瑞士乳杆菌(1株)。由于16S rDNA序列分析法不能区分植物乳杆菌和戊糖乳杆菌,为了进一步鉴定植物乳杆菌群中的4株菌,继续采用recA基因特异扩增和hsp60-RFLP技术对其细分,结果表明recA基因特异扩增和hsp60-RFLP的方法均能很好地把植物乳杆菌群中的4株菌鉴定到种水平,且均为植物乳杆菌。  相似文献   

16.
To simplify the labor-intensive conventional routine testing of samples to detect Leuconostoc at a meat processing plant, we developed polymerase chain reaction (PCR) primers specific for Leuconostoc from 16S rRNA gene sequences. These primers did not detect other common lactic acid bacteria such as Lactobacillus plantarum, Lact. sake, Lact. fermentum, Lact. acidophilus and Weissella viridescens. PCR with this primer detected all Leuconostoc species tested (Leu. mesenteroides subsp. mesenteroides, Leu. pseudomesenteroides, Leu. carnosum, Leu. lactic, Leu. citreum, Leu. amelibiosum, Leu. gelidum), except for Leu. fallax, and no other lactic acid bacteria on agarose gel electrophoresis. The method could identify areas contaminated with Leuconostoc in a large-scale industrial meat processing plant. Of 69 samples analyzed, 34 were positive for Leuconostoc according to the conventional culture method (isolation of LAB producing dextran) and PCR, whereas 29 were negative according to both. Six samples were culture-negative but positive by PCR. No false negative results were generated by PCR. The method is rapid and simple, is useful for routinely monitoring areas contaminated with Leuconostoc in meat processing plants, and could help to prevent the spoilage of meat products.  相似文献   

17.
Three beer‐spoilage strains of lactic acid bacteria (LAB), Lactobacillus brevis ABBC45, L. lindneri DSM 20690T and L. para‐collinoides DSM 15502T, exhibited strong ATP‐yielding ability in beer. To investigate energy sources, these beer‐spoilage strains were inoculated into beer. After the growth of the strains in beer, utilized components were determined by high performance liquid chromatography (HPLC). As a result, it was shown that citrate, pyruvate, malate and arginine were consumed by beer‐spoilage LAB strains examined in this study. The four components induced considerable ATP production even in the presence of hop compounds, accounting for the ATP‐yielding ability of the beer‐spoilage LAB strains observed in beer. We have further examined the ATP‐yielding ability of other strains of bacteria in beer. Beer‐spoilage bacteria, including Pectinatus frisingensis and P. cerevisiiphilus, showed strong ATP‐yielding abilities, whereas species frequently isolated from brewery environments exhibited low ATP‐yielding abilities. Although some of the nonspoilage LAB strains produced substantial amount of ATP in beer, the measurement of ATP‐yielding ability was considered to be useful as a rapid pre‐screening method for potential beer‐spoilage bacteria isolated from brewery environments.  相似文献   

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