软烤贻贝中蜡样芽孢杆菌生长/非生长界面模型建立与评价

建立软烤贻贝中蜡样芽孢杆菌标准菌株(ATCC49064与DSMZ 4312)在不同贮藏温度(T)、pH、水分活度(Aw)下生长/非生长界面模型,对其拟合情况和来自软烤贻贝蜡样芽孢杆菌(YB001)的验证情况进行分析和评价,并与已建立的脑心浸出液肉汤(BHI)中蜡样芽孢杆菌生长/非生长界面模型进行比较。所建模型总方程为Lopit(P)=-208.457-2.167·T+35.304·pH+705.573·Bw+1.117·T·pH-7.072·T·Bw-174.946·pH·Bw,其中R2-Nagelkerke=0.979和χ~2=0.019(df=8,P=1)显示拟合度较高,而且其预测一致率明显高于BHI培养基中建立的模型,表明该模型在预测软烤贻贝中蜡样芽孢杆菌的生长/非生长情况有很高的精确度和很好的适用性。此外,贮藏温度、水分活度、pH及其交互作用显著影响蜡样芽孢杆菌的生长/非生长(P0.05)。因此可以通过所建生长/非生长界面模型量化温度、水分活度、pH值等栅栏因子并结合其交互效应来确保软烤贻贝的高品质与安全性。     

软烤贻贝中蜡样芽孢杆菌生长/非生长界面模型的构建

基于不同贮藏温度、pH值和水分活度,使用R软件建立软烤贻贝中蜡样芽孢杆菌的生长/非生长界面logistic回归模型。分析结果得到,R2-Nagelkerke=0.979,Hosmer-Lemeshow检验中χ~2=0.0189,P=1,表明logistic回归模型拟合度高。此外,贮藏温度、水分活度、pH值及其交互作用对蜡样芽孢杆菌的生长/非生长情况影响显著(P0.05),并且贮藏温度、水分活度或pH值越低,蜡样芽孢杆菌受到的抑制越强。随着温度、水分活度或pH值的升高,它们对蜡样芽孢杆菌生长概率的主要影响因素地位被其余两个因素的交互作用所取代,而且交互作用中两因素的影响中一个增强,另一个减弱。建立的生长/非生长界面模型对实际生产有重要的指导意义,可以量化栅栏因子,并结合其交互效应,保证微生物的安全,及产品本身的营养和感官品质。     

环境因子交互作用下蜡样芽孢杆菌生长/非生长界面模型的建立与评价

本文旨在建立一种生长/非生长界面模型来预测蜡样芽胞杆菌在环境因子交互作用下的生长概率。选取五株蜡样芽孢杆菌菌株的混合菌株作为研究对象,研究温度、pH、Aw对混合菌株生长概率的交互影响。获得的生长/非生长实验数据用logistic回归方程拟合,建立了环境因子交互作用下蜡样芽孢杆菌生长/非生长界面模型。实验采取部分析因设计方案,选定80%的实验数据用做模型的拟合,20%的数据用做模型的验证。并从已发表的文献中选取30个数据作为测试集,通过比较预测值和观察值来检测已建模型的适用度。实验结果表明,训练集的一致性指数为0.991,验证集的一致率为0.988,说明模型对同类数据预测准确度高;同时模型的R2-Nagelkerke值也较高,为0.949;Hosmer-Lemeshow检验中的χ2=0.012,P=1,logistic回归模型拟合度较高。模型对测试集的预测准确率达83.3%,该模型对所选数据具有较高的预测能力,说明模型具有较广的适用范围。     

基于环介导等温扩增技术的产呕吐毒素 蜡样芽胞杆菌特异性检测

本研究旨在建立食品中产呕吐毒素蜡样芽胞杆菌的快速检测方法。基于产呕吐毒素蜡样芽胞杆菌合成酶基因cesB靶基因,设计4条特异性引物(2条内引物、2条外引物),建立环介导等温扩增技术(LAMP)。然后采用2株产呕吐毒素蜡样芽胞杆菌、19株蜡样芽胞杆菌和41株非蜡样芽胞杆菌验证了该LAMP具有很好的特异性。LAMP检测的灵敏度在DNA水平上达到1.49 pg/μL,纯菌的灵敏度为5×10~3 cfu/mL。人工污染米饭样品,当起始污染量为2 cfu/g时,37℃增菌培养6 h,用试剂盒法和煮沸法提取的DNA,都可以检出产呕吐毒素蜡样芽胞杆菌。采用此LAMP方法进行了72份实际样品的检测,检出2份产呕吐毒素蜡样芽胞杆菌阳性样品,与传统方法一致。因此,本研究建立的LAMP检测方法可应用于食品中产呕吐毒素蜡样芽胞杆菌的快速特异检测。     

动物性食品中蜡样芽胞杆菌的建模及风险评估应用进展

动物性食品有极高的营养价值,在食品组成中占有重要地位。蜡样芽胞杆菌是动物性食品中常见的一种食源性致病菌。综述了近年来国内外对蜡样芽胞杆菌的建模及在风险评估中的应用所开展的研究,对肉类、乳及乳制品、蛋及蛋制品等食物中建立的生长模型与失活模型的最新研究成果进行总结,概述了动态变化环境下的界面模型研究及模型在风险评估中的应用情况,最后对蜡样芽胞杆菌预测建模的研究及应用提出了展望,以期为今后的研究提供参考。     

蜡样芽胞杆菌生长特性及大蒜精油对其抑菌活性的研究

主要研究了37℃条件下3种蜡样芽胞杆菌在营养肉汤中的生长状况,建立了37℃下蜡样芽胞杆菌在营养肉汤中的Boltzmann牛长模犁,3条牛长曲线相关系数鼯均大于0．97;检测了不同培养时间蜡样芽胞杆菌的产芽胞情况,结果表明1号菌株和14号菌株较早产芽胞,培养相同时间,产芽胞数：1号菌株,4号菌株〉标准菌株;采用牛沣杯法和平板计数法研究了大蒜精油对蜡样芽胞杆菌的抑制效果,结果表明浓度为lO。的人蒜精油对3种蜡样芽胞杆菌都有很好的抑制效果,3种菌株中l号菌株最难抑制。     

婴幼儿奶粉和米粉中蜡样芽胞杆菌及其毒素、毒力基因的调查研究

了解婴幼儿奶粉及米粉中蜡样芽胞杆菌污染状况及其毒素、毒力基因的携带特点。方法 采用稀释培养计数(MPN计数)法分离蜡样芽胞杆菌,采用PCR技术检测10种蜡样芽胞杆菌的腹泻毒素及呕吐毒素基因,在流动相A为0.1%甲酸-乙腈溶液,流动相 B为0.1%甲酸-0.2 mmol/L乙酸铵溶液条件下,用Acquity BEH300 C₁₈色谱柱(100 mm×2.1 mm,1.7 μm)对样品进行分离,采用超高效液相色谱-串联质谱法检测样品中的呕吐毒素(cereulide)。结果 本研究共监测39份样品,28份检出蜡样芽胞杆菌,检出率为71.79%(28/39)；2份检出呕吐毒素,检出率为5.13%(2/39)。检出的蜡样芽胞杆菌菌株大多属于携带复合型毒素的菌株,均携带3种以上的腹泻毒素基因,非溶血性的肠毒素 nhe基因(nheA、nheB 和nheC)和肠毒素FM基因(entFM)为主要的毒力基因,其中nheABC 基因携带率为100%,entFM基因携带率为35.71%(10/28),cytK基因是检测到的最少的一种毒力基因。结论 应加强婴幼儿奶粉及米粉中的蜡样芽胞杆菌污染监测及其毒力基因致病性研究,以科学评估蜡样芽胞杆菌对婴幼儿食品可能构成的食品安全风险。     

乳酸对蜡样芽胞杆菌的抑制研究

本文研究了乳酸在营养肉汤培养基对蜡样芽胞杆菌的抑制,以及前期乳酸浸泡大豆后对蜡样芽胞杆菌在天培中生长情况的影响.研究发观,在营养肉汤培养基中,当采用乳酸调整pH时,在pH值为4.5时可以较好地抑制蜡样芽孢杆菌的生长.在天培的制作过程中,在前期未加乳酸浸泡处理的天培中,菌数可达108cfu/g并最终导致天培的酸败;在前期加乳酸浸泡处理过的天培中,蜡样芽胞杆菌亦可以生长,但菌数有所下降,为106cfu/g,乳酸浸泡对抑制天培中蜡样芽胞杆菌的生长起到一定的作用.     

量化栅栏技术和创建生长/非生长界面模型对食品保藏的意义

通过温度与水分活度(aw)和温度与pH对微生物生长速率联合作用的预测模型分析栅栏效应,由于食品中联合栅栏作用下,存在着微生物生长/非生长界面,探讨通过微生物生长动力模型来限定微生物生长/非生长界面的可能性。由于联合栅栏效应在不同的条件下,作用是不同的,有时是叠加的,有时是协同的。分析食品中栅栏作用就需要量化栅栏技术,量化栅栏技术可能为开发一种最低限度影响产品质量的新一代食品保藏技术提供一些新的思路。     

三因子金黄色葡萄球菌生长/非生长模型构建

运用统计分析R软件,建立金黄色葡萄球菌在温度、pH、盐浓度3种环境因子影响下的生长/非生长模型,构建的回归模型参数值a_1、a_2、a_3、a_4、a_5、a_6值均低于0.01,具有高度显著的拟合效果。分析各因子不同取值得到的方程式曲线图得出:在温度为10~25℃,pH值为4.5~5.5,盐浓度为0.5%~14.5%时,单因子对金黄色葡萄球菌生长/非生长影响显著;其他选值范围,3种因子协同影响。     

Survival and growth of foodborne pathogens during cooking and storage of oriental-style rice cakes

Fresh cooked rice cakes for retail sale are typically held at room temperature because refrigeration dramatically reduces their quality. Room temperature, high water activity, and a pH of > 4.6 provided an environment conducive to pathogen growth. To date, no studies have been published regarding survival and growth of foodborne pathogens in fresh cooked rice cakes. This study was undertaken to investigate the effect of steam cooking on foodborne pathogens and their subsequent growth in five varieties of rice cakes made from flours of regular rice, sweet rice, white rice, tapioca, and mung bean. Bacillus cereus spores were detected in white rice, tapioca, and mung bean samples. The rice cake flours were inoculated with non-spore-forming foodborne pathogens (Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, Listeria monocytogenes, and Staphylococcus aureus) or spore-forming bacteria (Bacillus cereus) and steam cooked (100 degrees C) for 30 min. Steam cooking significantly reduced (> 6 log CFU/g) non-spore-forming foodborne pathogens in all samples and inactivated spores of B. cereus by 1 to 2 log CFU/g. Although spores of B. cereus survived steam cooking and germinated during 3 days of storage at room temperature, populations in most rice cakes remained below 106 CFU/g, which is the threshold for producing toxin. Rice cakes made from mung bean flour supported growth and germination of B. cereus spores above that critical level. In mung bean rice cakes, enterotoxin production was detected by the second day, when B cereus cell populations reached about 6.9 log CFU/g. The toxin concentration increased with storage time. However, our results suggest that rapid growth of total mesophilic microorganisms by more than 7 to 8 log CFU/ml during the first day of storage produced off flavors and spoilage before B. cereus was able to grow enough to produce toxins. Therefore, steam-cooked rice cakes made from a variety of flours including mung bean flour are safe for sale for up to 1 day after storage at room temperature and are free of B. cereus toxins.     

响应面法优化蜡质芽孢杆菌低产正丙醇发酵工艺

为优化从浓香型白酒酿酒大曲和糟醅中分离出的低产正丙醇蜡质芽孢杆菌(Bacillus cereus)发酵工艺,本文通过单因素试验考察该菌发酵时间、接种量、发酵温度以及发酵液初始pH四个主要因素对发酵液中正丙醇含量的影响。然后以正丙醇含量为响应值,进行响应面分析和优化,建立了B. cereus发酵工艺的回归模型。结果表明,最佳发酵工艺为发酵时间33 h,接种量4.9%,发酵温度32 ℃,发酵液初始pH7.0。在此条件下,发酵液中正丙醇含量为(0.70±0.0012) mg/100 mL,和模型的预测值相吻合。     

Effect of nisin on growth boundaries of Listeria monocytogenes Scott A, at various temperatures, pH and water activities

The effect of nisin on growth boundaries of Listeria monocytogenes Scott A in Tryptone Soy Broth (TSB) under different a(w)s, pH, and temperatures was studied. Growth/no growth turbidity data was modeled using logistic regression. Combinations of various temperatures (5-35 degrees C), pH (4.05-6.70) adjusted with HCl, a(w)s (0.937-0.998) NaCl (0.5-10.5%) and nisin (0-100 IU/ml) were used to monitor the growth/no growth response of L. monocytogenes Scott A for 60 days. The concordance of the logistic regression model was 99.4%, indicating successful data fitting. The minimum pH at which growth was observed was 4.81 at the temperature range of 25-35 degrees C and at a(w) as high as 0.992. Growth was observed at a(w) as low as 0.937, at pH 6.7, at the temperature range of 25-35 degrees C. Increasing nisin concentrations above 25 IU/ml resulted in a more inhibitory environment for L. monocytogenes. Presence of 100 IU/ml resulted in a minimum pH for growth at 5.20, and a minimum a(w) at 0.967 at the temperature range of 25-35 degrees C. It was remarkable that low to medium salt concentrations (2.5-4.5 NaCl% w/v) provided a protective effect against inhibition of L. monocytogenes by nisin. The present study points out the applicability of growth/no growth modeling in order to study any interactions between various factors affecting initiation of growth of micro-organisms, in which its turn helps the understudying of microbe-food ecosystem relations and the development of safer food.     

豆瓣中腐败菌蜡样芽孢杆菌性能及对抗生素敏感性研究

该实验以豆瓣中分离的蜡样芽孢杆菌为实验菌株,对蜡样芽孢杆菌最适生长条件及对抗生素敏感性进行了检测。结果显示：该菌的最适生长温度是35 ℃,5 ℃以下和50 ℃以上不生长;最适生长盐含量为2%,在含量5%时生长缓慢,在盐含量10%以上时不生长;最适生长pH值为7.0,pH值3.0以下和pH值10.0以上该菌不生长;另外,在检测的25种抗生素中,有21种抗生素对该菌的生长具有抗性。     

Growth/no growth interfaces of Bacillus cereus, Staphylococcus aureus and Salmonella enteritidis in model systems based on water activity, pH, temperature and ethanol concentration

This study focuses its attention on the boundary between the growth and no growth of three strains ofSalmonella enteritidis , Bacillus cereus and Staphylococcus aureus in the presence of growth controlling factors such as temperature, pH, water activity (A_w) and ethanol concentration. Preliminarly, the minimal values of pH, A_wand temperature, and the maximum ethanol concentrations allowing the growth of the considered micro-organisms were determined. The calculation of these values enabled the use of logistic model to evaluate the growth/no growth boundary for the bacteria in relation to the considered independent variables. The location of the growth/no-growth boundaries for S. enteritidis and Staph. aureus were strongly affected, at the same ethanol concentration, by temperature, pH and A_w. Among the considered species, Staph. aureus was endowed with the highest sensitivity to low pH values whileB. cereus's growth/no growth interface, was quite unaffected by the combination of the stresses, when the physico–chemical conditions were above the minimum for growth. The effects of temperature, A_wand ethanol on the limitation of growth of the considered species were not merely additive. It was possible to identify the combinations of such factors preventing the growth of Salmonella enteritidis, Staph. aureus and B. cereus.     

Modeling the boundaries of growth of Salmonella Typhimurium in broth as a function of temperature, water activity, and pH

The growth limits of a mixture of five strains of Salmonella Typhimurium in tryptic soy broth were examined at different environmental conditions. The response of the pathogen was monitored in a total of 350 combination treatments of temperature (10 to 35 degrees C), pH (3.76 to 6.44), and water activity (aw, 0.913 to 0.990) for 62 days. No growth/growth (turbidity) data were modeled by logistic polynomial regression. The concordance index of the logistic model was 99.8%, indicating a good fit to the observed data. The minimum pH and aw values that permitted growth were 3.94 and 0.942, respectively, and occurred in the temperature range of 25 to 35 degrees C. At temperatures below this range, the minimum pH and aw allowing growth increased as the temperature decreased. The results showed an abrupt change in the probability of growth close to the boundary with minor changes of the environmental factors. The probabilities predicted by the model were compared with published data on the actual response of Salmonella Typhimurium or other salmonellae serotypes in 50 cases of food products, including salad dressing, mayonnaise, meat, cheese, vegetables, and fruits. The model predicted successfully the response of the pathogen in 90% of the tested cases. The results of the study indicated that the developed model predicts satisfactorily the growth/no growth interface of Salmonella Typhimurium in foods and can provide useful quantitative data for the development of safer food products and processes.     

THE NATURE OF TOXINS PRODUCED BY BACILLUS CEREUS BIS-59

Eight strains of Bacillus spp. were examined for production of toxin(s). Three strains of Bacillus cereus produced toxins lethal to mice. The culture filtrate of Bacillus cereus BIS-59 contained two lethal toxins. One was a hemolysin and the other a nonhemolytic glycoprotein. The growth of cells in fresh beef infusion broth resulted in high toxin production, compared to growth in tryptone-glucose-yeast extract or nutrient broth. Separate identities for the two toxins from B. vcereus BIS-59 were established on the basis of hemolytic activity, separation profiles on Sephadex gel filtration, responses to heat and radiation and kinetics of their production during the growth of the organism.     

The effect of calcium and sodium lactates on growth from spores of Bacillus cereus and Clostridium perfringens in a 'sous-vide' beef goulash under temperature abuse

The effect of calcium and sodium lactates on growth from spores of Bacillus cereus and Clostridium perfringens at three different concentrations (0, 1.5 and 3% w/w) and at different temperatures (10, 15 and 20 degrees C for B. cereus and 15, 20 and 25 degrees C for C. perfringens) was investigated, using beef goulash as a model system for pasteurised vacuum-packaged convenience foods. Calcium lactate at a level of 3% reduced the pH values of the samples from 6.0 to 5.5. No B. cereus growth was observed at 10 degrees C, but after 7 days at an incubation temperature of 15 degrees C, cell number increased by 1 log cfu/g in the control samples. At this temperature, lactates were seen to be effective at inhibiting growth. Calcium lactate was more inhibitory than sodium lactate as the growth of B. cereus was inhibited at 1.5 and 3% concentrations at 20 degrees C, respectively. Growth of C. perfringens was arrested in the presence of 1.5% calcium lactate at all storage temperatures, whereas growth was inhibited by 3% sodium lactate only at 15 degrees C.     

Effect of environmental parameters on growth kinetics of Bacillus cereus (ATCC 7004) after mild heat treatment

The influence of temperature (10 to 50 degrees C), initial pH (4.0 to 6.0) and sodium chloride concentration (0.5 to 3.0%) on the growth in nutrient broth and in meat extract of Bacillus cereus after mild heat treatment (90 degrees C--10 min) was determined. B. cereus spores survived after heating and they were able to germinate and grow in both media when post-treatment conditions were favourable. Heated B. cereus did not grow at 10 and 50 degrees C or in a medium with pH 4.0. Decreasing pH values and increasing levels of sodium chloride decreased growth rate and increased the lag phase of B. cereus. pH 4.5 was unable to prevent the growth of heated spores in a meat substrate with 0.5% NaCl at 12 degrees C. The combination of pH /=1.0% and temperatures /=60 degrees C could control heated B. cereus ATCC 7004 growth.     

Effect of temperature on growth characteristics of Bacillus cereus TZ415

The effect of temperature on the maximal specific growth rate was studied in Bacillus cereus between 5 and 40 degrees C cultivated in courgette broth and rich medium (J broth). B. cereus grown from 5 to 38 degrees C in rich medium. No growth was observed in courgette broth below 10 degrees C. The Arrhenius plot was fitted from experimental data of B. cereus grown in rich medium and at regulated pH, oxygen and temperature. Two domains which are separated by a critical temperature around 13 degrees C can be distinguished with regard to temperature dependence of maximal specific growth rate. Over the cold domain from 5 to 13 degrees C, the temperature characteristic was 2.6 fold higher than over the sub-optimal domain from 13 to 38 degrees C suggesting that the growth temperature regulates several metabolic pathways.