响应面法优化啤酒有害菌检测培养基

为了提高啤酒有害菌的检出率和检测的灵敏性,采用Plackett-Burman设计法和响应面分析法(Response Surface Analysis)对CNFF-A培养基组分和配比进行优化。先用Plackett-Burman设计从7个因子中筛选出对菌落数有显著影响的因素,再用最陡爬坡试验及Box-Behnken设计进一步优化。结果表明,蜂蜜、叶酸和乙酸钠是影响菌落数的显著因素,优化后的培养基配方为:蜂蜜0.1g/L、牛肉浸粉7.0g/L、酵母浸出物5.0g/L、精氨酸1.0g/L、叶酸2.0g/L和乙酸钠1.5g/L。此条件下,培养基对啤酒有害菌的检出率大大提高,且检测时间缩短了24h。     

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     

新型酒花制品在啤酒工业的应用

啤酒花是啤酒工业的重要原料之一。阐述了啤酒花中3类物质即酒花树脂、酒花油、多酚物质的主要化学成分，酒花在啤酒酿造中的作用，以及酒花浸膏和酒花油等多种新型的酒花制品在啤酒工业中的应用。     

The influence of thiamine and riboflavin on various spoilage microorganisms commonly found in beer

Beer is generally considered a stable product owing to its intrinsic ‘unfavourable’ conditions (hops, alcohol, low oxygen, etc.) that inhibit the growth of pathogenic microorganisms. However spoilage microorganism such as Lactobacillus brevis , Pediococcus damnosus , Acetobacter aceti , Zymomonas mobilis and various wild yeasts (e.g. Brettanomcyes spp.) can have significant detrimental effects on the organoleptic properties of the final product. The presence of essential vitamins, such as thiamine and riboflavin, can help to enhance the growth of these microorganisms, accelerating the rate of spoilage. The presence of thiamine had a noticeable effect on the lactic acid productivity of L. brevis and P. damnosus , acetaldehyde productivity of Z. mobilis and acetic acid production of Brettanomyces spp., while riboflavin enhanced 2,3‐pentanedione production by P. damnosus and Brettanomyces spp. Copyright © 2017 The Institute of Brewing & Distilling     

食源性腐败菌挥发性有机化合物的研究进展

食源性腐败菌是食品腐败的重要原因, 它可通过初级或次级代谢产生具有异味的挥发性有机化合物(volatile organic compounds, VOCs), 加速食品的腐败进程。腐败菌种类及组成会随食品基质和贮藏条件而改变, 导致产生的VOCs也具有多样性和差异性。随挥发性有机物质收集和检测技术的快速发展, 通过分析腐败菌相关的VOCs成分组成可鉴别食源性腐败菌种类和食物腐败进程。本文对常见腐败菌VOCs的产生途径和种类进行归纳, 并重点探讨了VOCs提取分析技术及在食品腐败研究中的具体应用, 为有效进行腐败菌防控, 提升食品质量和安全提供参考。     

冷鲜牛肉的优势腐败菌及其消长规律

冷鲜牛肉的腐败与微生物的种类和数量密切相关。对冷鲜牛肉的初始菌相和不同贮藏温度条件下菌相及pH的变化规律进行研究,结果表明:假单胞菌和乳酸菌是冷鲜牛肉初始菌相中的主要微生物;假单胞菌和热死环丝菌是冷鲜牛肉的优势腐败菌;不同贮藏温度下细菌总数达到107 cfu/g的时间为2～8 d,pH的变化范围为5.6～6.8。     

海水鱼优势腐败菌腐败能力分析

研究大黄鱼和大菱鲆无菌鱼块接种优势腐败菌后5℃贮藏中的感官、腐败产物和腐败菌的变化,以生长动力学参数和腐败产物的产量因子(YTVBN/CFU和YTMA/CFU)为指标,探讨两种冷藏海水鱼优势腐败菌希瓦氏菌和假单胞菌的腐败能力。结果表明,大菱鲆鱼块接种腐败希瓦氏菌和恶臭假单胞菌的货架期分别为60,72h,货架期终点时的TVBN含量分别为35.48,37.56mg/100g,腐败菌菌数分别为8.14,8.32lg(CFU/g),产量因子YTVBN/CFU分别为1.86×10-7,1.35×10-7 mg TVBN/CFU。大黄鱼鱼块接种腐败希瓦氏菌和荧光假单胞菌的货架期分别为162,174h,货架期终点时的TVBN含量分别为31.74,39.01mg/100g,腐败菌菌数分别为8.71,8.91lg(CFU/g),产量因子YTVBN/CFU分别为4.49×10-8,3.72×10-8 mg TVBN/CFU。大黄鱼鱼块的货架期比大菱鲆的明显长,接种假单胞菌的两种鱼块的货架期比接种希瓦氏菌的稍长。两种海水鱼低温有氧贮藏优势腐败菌希瓦氏菌和假单胞菌都有很强的腐败能力。     

影响啤酒质量的因素及其检验

总结了影响啤酒质量的几种主要因素,介绍了监检这些因素的方法,对提高啤酒质量有一定的参考价值。     

凡纳滨对虾优势腐败菌鉴定及其致腐能力的初步研究

分析鉴定了凡纳滨对虾0℃与20℃贮藏条件下的菌相组成与优势腐败菌,并对优势腐败菌16SrDNA、生长动力学、致腐能力与菌落数的变化进行了测定。结果表明,0℃与20℃贮藏条件下,对虾优势腐败菌分别是希瓦氏菌(30%)、不动杆菌(16.7%)与希瓦氏菌(46.5%)、发光杆菌(17.7%)。7℃条件下,将一定浓度的希瓦氏菌与不动杆菌菌悬液接种到无菌对虾上,结果显示接种希瓦氏菌的样品其腐败代谢产物产量因子YTVB-N/CFU、YTMA/CFU分别为12.44×10-9、6.193×10-10,而接种不动杆菌的样品其YTVB-N/CFU、YTMA/CFU分别为8.937×10-9、5.548×10-10。结果表明,7℃条件下,希瓦氏菌的致腐能力强于不动杆菌,希瓦氏菌在对虾腐败过程中占主导作用,其分析结果与对虾菌相组成的鉴定结果相一致。     

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     

食源性致病菌和腐败菌的快速检测方法研究进展

食源性致病菌和腐败菌污染一直是引发食品安全事件的重要因素。传统的检测方法虽具有较高的准确性,但需培养及生化实验,耗费时间长且操作较复杂,开发高效、快速的检测方法对保障食品安全至关重要。近年来,科学技术的进步使一些新方法、新技术逐渐被运用,比传统培养法检测时间更短、效率更高、特异性更好,具有广阔的应用前景。因此,本文综述了食源性致病菌和腐败菌快速检测方法的最新研究进展,包括纸片法、流式细胞术、阻抗法、腺苷三磷酸荧光法、光谱检测技术、分子生物学检测技术、免疫学检测法及生物传感器检测技术等,着重论述了各种方法的原理、优缺点、应用状况及发展方向,为致病菌和腐败菌引发的食品安全事件的预防及控制提供参考。     

冰温贮藏中南美白对虾特定腐败菌的分离鉴定及其腐败能力分析

为了分离鉴定南美白对虾在冰温贮藏条件下的腐败菌,并分析其腐败能力的大小。通过冻结实验,确定南美白对虾的冰点,在0℃与冰点之间贮藏南美白对虾,从中分离腐败菌,进行形态学观察和分子生物学鉴定,将分离得到的腐败菌接种至灭菌虾汁中,通过产硫化氢,降解蛋白质,产挥发性盐基氮（TVB-N）的测定衡量腐败能力的大小。结果表明:从南美白对虾中分离得到12株腐败菌,分别是Shewanella hafniensis,Acinetobacter johnsonii,Planoccoccus citreus,Bacillus cereus,Acinetobacter beijerinckii,Enterobacter hormaechei,Arthrobacter bergeri,Bacillus licheniformis,未鉴定出的菌株X1和X10。在冰温贮藏条件下,导致南美白对虾腐败的优势腐败菌依次是Shewanella hafniensis、X1、Acinetobacter johnsonii和Acinetobacter beijerinckii。      

High‐pressure homogenization: a non‐thermal process applied for inactivation of spoilage microorganisms in beer

The inactivation of spoilage microorganisms in beer using high‐pressure homogenization (HPH) was studied with the aim of evaluating the possibility of changing the conventional pasteurization process using this particular process. The homogenization pressure required for the inactivation of lactic acid bacteria, acetic bacteria and yeasts was investigated. For the most resistant microorganisms, the pressure inactivation kinetics and the effects of multiple process passes, initial temperature of the beer and the CO₂ concentration were studied. The results indicated that Lactobacillus delbrueckii was the most resistant microorganism tested, requiring 250 MPa to reach a six decimal reduction. Additionally, results showed that L. delbrueckii inactivation followed a second‐order kinetic process. A multi‐pass process and the use of a high initial beer temperature increased inactivation by HPH with L. delbrueckii, allowing the use of 150 MPa to achieve a five log cycle of inactivation. In contrast, a high CO₂ concentration reduced the efficacy of the HPH process. The results that were obtained are useful for high‐pressure homogenization applications in breweries and help to elucidate the effect of this new technology in a beverage that is both alcoholic and carbonated. Copyright © 2013 The Institute of Brewing & Distilling     

冷却猪肉中优势腐败菌致腐能力研究

为明确冷却猪肉中优势腐败菌对其腐败的作用程度,本研究监测冷却猪肉冷藏过程中菌落总数和优势腐败菌的变化情况,并接种假单胞菌、气单胞菌、肠杆菌和热杀索丝菌于冷却猪肉中,分组检测尸胺含量在冷却猪肉冷藏过程中的变化。实验表明,优势腐败菌在冷却猪肉冷藏第5d后显著上升(p<0.05),与尸胺含量变化趋势相似,同时接种多种腐败菌与尸胺含量极显著相关(p<0.01),表明优势腐败菌的协同效应对冷却猪肉腐败具有重要作用。     

Influence of hop harvest date of the ‘Mandarina Bavaria’ hop variety on the sensory evaluation of dry‐hopped top‐fermented beer

To impart a special hop aroma to beer, dry‐hopping is a technique that is becoming more and more popular with commercial breweries. Nevertheless, until now little was known about the factors that influence the reproducibility (and consistent product quality) of dry‐hopping with flavour varieties. One factor that could influence the sensory impressions and aroma profile compositions of dry‐hopped beers is the hop harvest date. Therefore, to determine the effects of different harvest dates of the flavour variety ‘Mandarina Bavaria’ on the aroma of top‐fermented beer, laboratory‐scale dry‐hopping trials were performed. Besides tasting sessions of brewed beers, relative quantities of selected hop‐derived, as well as beer‐originated aroma compounds, were investigated by headspace–solid‐phase microextraction–gas chromatography–mass spectrometry. Duo–trio tests between the beers hopped with pellets of different harvest dates showed no significant differences (α = 0.05) between them. In addition, these beers had similar profiles in a five‐point profile tasting scheme. On the other hand, relative concentrations of some hop‐derived aroma compounds – especially myrcene, which is known to be able to contribute to beer flavour – increased corresponding to a later harvest date, while beer originated volatiles were not different between the beers. Analytical results combined with the results of sensory evaluations led to the conclusion that the harvest date of Mandarina Bavaria was not a dominant factor in the dry‐hopping aroma of top‐fermented beers. High amounts of fermentation by‐products are likely responsible for masking effects resulting in no sensory distinctness between the samples with different hop aroma compound concentrations. Copyright © 2016 The Institute of Brewing & Distilling     

不同包装条件下冷却肉品质变化及腐败菌相研究

为寻找冷却猪肉合理的包装方式,延长货架期,将猪后腿肉分别进行托盘包装、真空包装、高氧气调包装（20%CO₂+80%O₂）和低氧气调包装（65%CO₂+35%O₂）处理,并以无包装组为对照,测定样品在4℃贮藏过程中挥发性盐基氮（TVB-N）值、色差值a*、丙二醛（MDA）值、微生物菌落（菌落总数、假单胞菌菌落数和热杀索丝菌菌落数）和感官评分。结果表明:无包装和托盘包装冷却猪肉货架期不超过4 d,高氧气调包装（20%CO₂+80%O₂）货架期为6 d,真空包装和低氧气调包装（65%CO₂+35%O₂）货架期长达9 d。真空和气调包装均有利于延长冷藏猪肉货架期,但真空包装使肉呈暗红色。气调包装中O₂和CO₂的含量影响肉的品质,高氧气调包装有利于好氧菌假单胞菌和热杀索丝菌的生长繁殖,同时促进脂质氧化,但可以很好维持肉色,低氧气调包装（65%CO₂+35%O₂）可以抑制细菌的生长和脂肪氧化。综合各项评判结果,得到最佳包装方式为低氧包装（65%CO₂+35%O₂）。      

Effect of bacteriocin‐producing Pediococcus acidilactici K10 on beer fermentation

Beer is generally considered to be a beverage that has high microbiological stability. However, some undesirable lactic acid bacteria (LAB) can grow in beer and consequently spoil this beverage. In this study, bacteriocin‐producing Pediococcus acidilactici K10 was used as a means of bio‐acidifying the mash and reducing the spoilage LAB content of the beer. The K10 strain had antimicrobial activity against two beer spoilage LAB strains in wort and did not grow in a beer environment. The K10 strain was inoculated before the mashing step. The effect of K10 as a starter culture was investigated and compared with a control. As a result, filtration time was shortened by 17 min, alcohol content was increased by 137%, foam stability was increased by 156%, bitterness was increased by two bitterness units and there was a significant difference (p < 0.05) in aromatic and sour odour. The feasibility of using bacteriocin‐producing LAB strain in beer brewing is envisaged. Copyright © 2016 The Institute of Brewing & Distilling     

Possibilities to improve the antioxidative capacity of beer by optimized hopping regimes

Different hopping regimes were evaluated to investigate the effect on the oxidative stability of wort and beer. Compared with a single hop dosage at the beginning of wort boil, it was possible to increase the concentration of α‐acids in pitching wort and beer by applying incremental hop dosage, dry hopping or the use of a pre‐isomerized hop product in combination with an α‐acid extract, which concomitantly resulted in lower iron concentrations and an enhanced flavour stability as indicated by standard wort and beer analyses, atomic absorption spectroscopy, electron spin resonance spectroscopy and sensory analysis of fresh and force‐aged beers. The functional principle of hop dosage variations is explained by saving of α‐acids throughout the wort production process, which yields an increased formation and precipitation of pro‐oxidative acting transition metal ions (e.g. Fe) in α‐acid‐complexes during the whirlpool rest and fermentation. Consequently, fewer reactive oxygen species are generated. Additional laboratory trials simulating wort cooling and beer storage in buffered model solutions proved that un‐isomerized α‐acids are strong iron chelators and confirmed the functional principle of the applied hopping regimes. Negative effects of higher α‐acid contents on fermentation performance and depletion of the zinc concentration, which is an essential nutrient for yeast, could be excluded. Copyright © 2014 The Institute of Brewing & Distilling     

应用PCR-DGGE对巴氏消毒乳中腐败细菌的污染溯源研究

运用PCR-DGGE技术分析了巴氏杀菌乳从原料乳到杀菌、灌装这两个加工关键控制点的微生物动态变化和差异性,容易发生污染的环节是挤乳环节、杀菌后的加工管道和包装环节。原料乳中易携带的细菌是乳酸链球菌（Lactococcus lactis subsp.）和乳酸杆菌（Lactobacillus sp.）;挤乳环节中易受无乳链球菌（Streptococcus agalactiae）污染;杀菌后的加工管道易受热杀环丝菌（Brochothrix thermosphacta）、钻黄肠球菌（Enterococcus casseliflavus）、瑞士乳杆菌（Lactobacillus helveticus）和2株非培养的假单胞菌属（Uncultured Pseudomonas sp.）污染;肉杆菌属（Carnobacterium sp.）和产乳酸菌素的肉杆菌（Carnobacterium maltaromaticum）则在挤乳环节、杀菌后的加工管道或包装环节都可能被污染。