Predicting microbial heat inactivation under nonisothermal treatments

The aim of this study was to develop an equation that accurately predicts microbial heat inactivation under nonisothermal treatments at constantly rising heating rates (from 0.5 to 5 degrees C/min) in media with different pH values (4.0 or 7.4). The survival curves of all bacteria (Enterococcus faecium, Escherichia coli, Listeria monocytogenes, Salmonella Senftenberg 775W, Salmonella Typhimurium, and Staphylococcus aureus) tested under isothermal treatments were nearly linear. For the most heat-resistant microorganism (E. faecium), the estimated DT-values at pH 7.4 were at least 100 times those of the second most thermotolerant microorganism (Salmonella Senftenberg 775W). The heat resistance of E. faecium was up to 30 times lower at pH 4.0 than at pH 7.4. However, E. faecium was still the most heat-resistant microorganism under nonisothermal treatments at both pH values. Inactivation under nonisothermal conditions was not accurately estimated from heat resistance parameters of isothermal treatments when microbial adaptation or sensibilization occurred during the heating up lag phases. The under-prediction of the number of survivors might be greater than 15 log CFU within the nonisothermal treatment conditions investigated. Therefore, the nonisothermal survival curves of the most heat-resistant microorganisms were fitted with the following equation: log S(t) = -(t/delta)P. This equation accurately described the survival curves of all the bacteria tested. We observed a linear relationship between the log of the scale parameter (delta) and the log of the heating rate. A p value characteristic of each microorganism and pH tested was calculated. Two equations capable of predicting the inactivation rate of all bacteria tested under nonisothermal treatments at pH 7.4, 5.5, or 4.0 were developed. The model was evaluated in skim milk and apple juice. The results of this study could be used to help minimize public health risks and to extend the shelf life of those foods requiring long heating up lag phases during processing.     

不同温度培养诱导金黄色葡萄球菌对超高压失活的抗性及其建模

探讨不同培养温度处理诱导金黄色葡萄球菌(ATCC 6538)对超高压的抗性,并建立不同培养温度下金黄色葡萄球菌的超高压抗性模型。金黄色葡萄球菌经不同温度培养,在100~500 MPa的超高压条件下,选用线性、Weibull和Gompertz三种模型来拟合超高压抗性曲线,以决定系数(R2),均方误差(RMSE),精确因子(Af)和偏差因子(Bf)作为模型拟合度优劣的评判指标。实验结果表明,在100~500 MPa压力的作用下,线性模型的拟合效果不佳,R2最小值达到0.8870,Weibull和Gompertz模型对超高压的抗性具有较好的拟合性(R2≥0.9467),且Weibull模型的拟合效果最好,R2最大值达到0.9956,RMSE最小值为0.0312。因此,Weibull模型可以很好地拟合金黄色葡萄球菌以不同的培养温度胁迫后在超高压作用下的抗性曲线,随着培养温度的升高,金黄色葡萄球菌的超高压抗性呈增加趋势。      

Comparing predicting models for heat inactivation of Listeria monocytogenes and Pseudomonas aeruginosa at different pH

Under the same experimental conditions it has been demonstrated that whereas survival curves of Listeria monocytogenes in the range of temperatures from 54 to 62 °C followed a first-order kinetic, those of Pseudomonas aeruginosa in the range of temperatures from 50 to 56 °C were not linear showing a shoulder followed by a linear region. The first order kinetic model did not describe survival curves of P. aeruginosa. A model based on the Weibull distribution (Log₁₀(N_t/N₀)=(1/−2.303)*(t/b)ⁿ)) accurately described the inactivation kinetics of both microorganisms at the three pHs of 4, 5.5, 7.4 investigated. For both microorganisms, the b value depended on the treatment temperature and the pH of the treatment medium. Whereas for L. monocytogenes the n value was independent of the treatment conditions, for P. aeruginosa the n value depended on the pH of the treatment medium.

The model based on the Weibull distribution was capable of accurately predicting the treatment time to inactivate five Log₁₀ cycles of both microorganisms at the three pHs investigated.     

Predicting heat inactivation of Listeria monocytogenes under nonisothermal treatments

The aim of this study was to find a model that accurately predicts the heat inactivation of Listeria monocytogenes (ATCC 15313) at constantly rising heating rates (0.5 to 9 degrees C/min) in media of different pH values (4.0 to 7.4). Survival curves of L. monocytogenes obtained under isothermal treatments at any temperature were nearly linear. Estimations of survival curves under nonisothermal treatments obtained from heat resistance parameters of isothermal treatments adequately fit experimental values obtained at pH 4.0. On the contrary, survivors were much higher than estimations at pH 5.5 and 7.4. The slower the heating rate and the longer the treatment time, the greater the differences between the experimental and estimated values. An equation based on the Weibullian-like distribution, log S(t) = (t/delta)p, accurately described survival curves of L. monocytogenes obtained under nonisothermal conditions within the range of heating rates investigated. A nonlinear relationship was observed between the scale parameter (delta) and the heating rate, which allowed the development of an equation capable of predicting the inactivation rate of L. monocytogenes under nonisothermal treatments at pH 5.5 and 7.4. The model predictions were a good fit to the measured data independent of the magnitude of the thermotolerance increase. This work might contribute to the increase in safety of those food products that require long heating lag phases during the pasteurization process.     

脉冲强光对馒头表面金黄色葡萄球菌的杀菌效果

目的研究脉冲强光技术在馒头表面金黄色葡萄球菌杀菌中的效果及其机制。方法控制闪照距离为9~21 cm,单脉冲能量为100~500 J,闪照次数为0~20次,分析不同条件下金黄色葡萄球菌的失活规律,并将该优化条件应用于馒头表面金黄色葡萄球菌的杀菌试验,最后采用透射电子显微镜技术分析脉冲强光技术对金黄色葡萄球菌形态结构的影响,从而分析其杀菌机制。结果脉冲强光在闪照距离为9 cm,单脉冲能量为500 J的情况下,闪照16次能对金黄色葡萄球菌产生良好的杀菌效果。该脉冲强光处理技术能降低馒头表面的金黄色葡萄球菌1.72 log CFU/g。透射电镜观察结果说明了脉冲强光可破坏金黄色葡萄球菌的细胞膜和菌体胞质内容物,从而造成细胞的失活。结论脉冲强光可对金黄色葡萄球菌产生较强的杀菌效果,在馒头加工业中具有一定的应用前景。     

光动力对金黄色葡萄球菌的杀伤作用及其AFM观察

通过细菌菌落计数以及原子力显微镜图像研究了血啉甲醚对金黄色葡萄球菌的光动力杀伤作用。结果表明:HMME在金黄色葡萄球菌菌液中的最强吸收峰为392nm,溴钨灯光源波长范围350～2500nm,能满足HMME所需的波长;HMME浓度为5μg/mL接受溴钨灯光源光照30min能杀死82.2%的金黄色葡萄球菌;原子力显微镜图像显示其灭菌机制是HMME在光照条件下导致菌体破裂,细胞内容物大量外泄;无光照条件下,HMME对金黄色葡萄球菌杀伤效果不显著。     

Comparing predicting models for heat inactivation of Listeria monocytogenes and Pseudomonas aeruginosa at different pH

Under the same experimental conditions it has been demonstrated that whereas survival curves of Listeria monocytogenes in the range of temperatures from 54 to 62 °C followed a first-order kinetic, those of Pseudomonas aeruginosa in the range of temperatures from 50 to 56 °C were not linear showing a shoulder followed by a linear region. The first order kinetic model did not describe survival curves of P. aeruginosa. A model based on the Weibull distribution (Log₁₀(N_t/N₀)=(1/−2.303)*(t/b)ⁿ)) accurately described the inactivation kinetics of both microorganisms at the three pHs of 4, 5.5, 7.4 investigated. For both microorganisms, the b value depended on the treatment temperature and the pH of the treatment medium. Whereas for L. monocytogenes the n value was independent of the treatment conditions, for P. aeruginosa the n value depended on the pH of the treatment medium.The model based on the Weibull distribution was capable of accurately predicting the treatment time to inactivate five Log₁₀ cycles of both microorganisms at the three pHs investigated.     

Inactivation of Staphylococcus aureus in milk using flow-through pulsed UV-light treatment system

ABSTRACT:  This study investigated the efficacy of pulsed UV-light for continuous-flow milk treatment for the inactivation of Staphylococcus aureus , a pathogenic microorganism frequently associated with milk safety concerns. Pulsed UV light is an emerging technology, which can be used for the inactivation of this pathogen in milk in a relatively short time. Pulsed UV light damages the DNA of the bacteria by forming thymine dimers that lead to bacterial death. The effect of sample distance from the quartz window of the UV-light source, number of passes, and flow rate was investigated. A response surface methodology was used for the design and analysis of experiments. Milk was treated at 5-, 8-, or 11-cm distance from a UV-light strobe at 20, 30, or 40 mL/min flow rate and treated up to 3 times by recirculation of milk to assess the effect of the number of passes on inactivation efficiency. Log₁₀ reductions varied from 0.55- to 7.26-log₁₀ CFU/mL. Complete inactivation was obtained in 2 cases and no growth was observed following an enrichment protocol. Predicted results were in agreement with the experimental data. Overall, this work demonstrates that pulsed UV-light has a potential for inactivation of milk pathogens.     

金黄色葡萄球菌在生乳中生长预测模型的建立

目的建立生乳中金黄色葡萄球菌的生长预测模型。方法采用GB/T4789.37—2008第三法,测定不同温度下金黄色葡萄球菌在生乳中的生长曲线,利用麦夸特法和通用全局优化法将曲线数据分别拟合Gompertz方程、Logistic方程、Richards方程、Weibull方程、Baranyi方程和Monod方程,确定最适方程,建立一级模型。同时应用响应面方程建立二级生长预测模型。结果金黄色葡萄球菌在生乳中的最适预测模型为Gompertz模型。结论通过与多模型比较,建立了金黄色葡萄球菌在生乳中的生长预测模型。为该菌在生乳中的风险评估理论体系的建立提供理论基础,也为乳制品的安全生产和流通提供了科学依据。     

微波加热对三种微生物致死的Weibull模型

探讨不同微波功率加热至不同温度下对沙门氏菌、大肠杆菌和金黄色葡萄球菌等三种对象菌的致死作用,运用Weibull模型描述其致死历程,由Weibull模型计算微波加热使微生物数量减少5-log所需要的时间,并验证其模型的精确度。结果表明,Weibull模型描述沙门氏菌、大肠杆菌和金黄色葡萄球菌的致死历程均有较高的拟合度,通过Weibull模型推算出来的沙门氏菌、大肠杆菌和金黄色葡萄球菌减少5-log的时间与实验结果相近。     

食品加工中的减菌处理对残存金黄色葡萄球菌生长的影响研究

为了解食品加工过程中的减菌处理对残存微生物生长的影响。本文以金黄色葡萄球菌CMCC（B）26003为实验菌株,先将培养至对数期的菌液经过加热、冷冻、紫外线照射、次氯酸钠溶液浸泡、臭氧水浸泡处理后,接种至液体培养基中37℃下振荡培养12～14h,定时取样测定菌液浓度,Baranyi方程构建生长模型,根据生长迟滞期和最大比生长速率大小分析各种减菌处理方法对残存金黄色葡萄球菌生长的影响。结果表明,60℃加热5min和30mg/kg次氯酸钠溶液浸泡10min两种处理对金黄色葡萄球菌的生长迟滞期影响显著（p<0.01）,分别为3.95h和2.62h,约为对照的2.6和2.0倍。60℃加热5min和18W紫外照射15min两种处理对金黄色葡萄球菌的最大比生长速率影响显著（p<0.01）,分别为1.81logcfu/mL.h^-1和1.83logcfu/mL.h^-1,约为对照的1.1倍。0.5mg/kg臭氧水浸泡10min和-18℃冷冻3d两种处理对金黄色葡萄球菌的生长动力学参数影响不大。      

1874株金黄色葡萄球菌临床分布及耐药性结果分析

目的了解四川省某医院甲氧西林敏感金黄色葡萄球菌(methicillin sensitive Staphylococcus aureus,MSSA)与耐甲氧西林金黄色葡萄球菌(methicillin resistant Staphylococcus aureus,MRSA)的临床科室分布和耐药性情况。方法回顾性分析2014年12月-2017年12月四川省某医院门诊及住院患者,包括8名因金黄色葡萄球菌引起的食物中毒患者,送检各类标本分离出SA(Staphylococcus aureus)可疑菌落1874株,对其进行鉴定和药敏检测。结果 MSSA和MRSA标本主要来源于痰液(51.31%,74.33%)、伤口分泌物(21.52%,8.96%)和脓液(10.07%,7.92%)。725株MSSA临床分离率依次为门诊(27.17%)、呼吸科(15.45%)、ICU(10.21%)、神经科(8.55%)和口腔科(5.52%),1149株MRSA依次为ICU(33.16%)、神经科(11.66%)、呼吸科(9.75%)、儿科(7.57%)和妇产科(6.27%)。耐药性结果表明,MSSA对苯唑西林、万古霉素、利奈唑胺、替考拉林敏感,对青霉素G、红霉素和阿莫西林/克拉维酸耐药率依次为89.38%、56.69%、21.51%外,对其他抗菌药耐药率均18.00%; MRSA对万古霉素利、奈唑胺、替考拉林敏感,对苯唑西林、莫西沙星、克林霉素、红霉素、青霉素G、四环素、环丙沙星、阿莫西林/克拉维酸和氨苄西林/舒巴坦的耐药率均显著高于MSSA。结论四川省某医院2014年12月-2017年12月期间临床分离的SA以MRSA为主, MSSA和MRSA在标本来源、科室分布等方面存在较大差异, MRSA耐药情况更为严重,明显高于MSSA。若要评估金黄色葡萄球菌感染风险以及病人的治疗方案,应分别考虑MRSA和MSSA的感染来源和耐药性。     

香茅醛对金黄色葡萄球菌的抑制作用

该研究探讨香茅醛（Citronellal,CIT）对金黄色葡萄球菌（Staphylococcus aureus,S. aureus）的抑菌作用。采用微量肉汤稀释法及时间-杀菌曲线确定了CIT对S.aureus的抑菌活性。CIT对S.aureus最小抑菌浓度（MIC）及最小杀菌浓度（MBC）均为0.80 mg/mL,且具有快速杀菌作用,1.60 mg/mL的CIT处理0.5 h后,S. aureus活菌数从5.65 log CFU/mL下降到0.00。经CIT处理后,S. aureus胞外K+浓度显著升高至2.63 mg/L,碘化丙啶（PI）荧光强度与对照相比升高5 326.83 A.U.,表明CIT会影响细胞膜通透性,导致胞内小分子及大分子物质泄出。扫描电镜观察下发现,高浓度CIT能严重破坏S.aureus细胞形态及结构。此外,通过紫外可见光光谱发现,CIT在加入S. aureus的DNA后,吸收峰发生明显的增色效应,与对照组相比最大差值为0.45 A.U.,表明CIT可能通过沟槽结合与DNA相互作用,干扰DNA的正常生物功能,抑制细胞生长。综上所述,CIT可作为S. aureus生长...     

应用基因芯片技术鉴定食源性金黄色葡萄球菌的研究

目的:建立快速鉴定食源性金黄色葡萄球菌的基因芯片技术。方法:采用PCR方法扩增金黄色葡萄球菌16SrRNA基因的DNA片段,序列进行BLAST比较并通过软件设计其特异性探针,采用基因芯片杂交技术鉴定食源性金黄色葡萄球菌样品。结果:对所有样品进行基因芯片杂交技术处理并扫描观察,金黄色葡萄球菌的杂交结果呈阳性,其检测结果与传统方法鉴定结果一致。结论:应用基因芯片鉴定技术可快速、准确地鉴定食源性金黄色葡萄球菌,值得推广应用。     

食品中金黄色葡萄球菌PCR-ELISA检测技术建立

针对食品中金黄色葡萄球菌检测,选取金黄色葡萄球菌耐热核酸酶nuc基因作为靶标,利用地高辛标记引物扩增的PCR产物与生物素标记的探针杂交,然后与ELISA平板上的链霉素杂交,通过抗地高辛抗体显色建立PCR-ELISA检测技术。应用该方法检测人工污染牛奶中金黄色葡萄球菌,同时将大肠埃希氏菌、沙门氏菌、志贺氏菌、单增李斯特菌等常见食源性致病菌作为阴性对照。结果显示,金黄色葡萄球菌的纯培养物以及人工污染金黄色葡萄球菌牛奶,金黄色葡萄球菌为阳性,其他对照菌为阴性,两者PCR-ELISA检测敏感性均为10¹CFU/m L,比普通PCR检测的敏感性（10³CFU/m L）高100倍。因此,该研究建立的PCR-ELISA方法具有良好的特异性与敏感性,能够特异、敏感、准确地检测牛奶中的金黄色葡萄球菌,为食品中金黄色葡萄菌的快速鉴定、风险评估与有效监测提供重要技术手段。      

鉴定金黄色葡萄球菌的多重PCR方法

建立了一种快速鉴定和鉴别金黄色葡萄球菌的多重PCR方法。利用金黄色葡萄球菌的特异基因、耐甲氧西林基因和4种肠毒素基因nuc、mecA和sea、sec、sed、see建立6重PCR反应体系,并对PCR体系,引物浓度进行优化。6对PCR引物均能特异地扩出相应的目的基因。对47株金黄色葡萄球菌的检测结果与应用单重PCR鉴定结果完全一致。实验所建立的多重PCR方法能在一个反应体系中鉴定和鉴别金黄色葡萄球菌,为食品中金黄色葡萄球菌的快速检测提供了一种有效的检测手段。     

碳纳米生物传感检测金黄色葡萄球菌

目的建立一种基于碳纳米材料的适配体传感器快速检测金黄色葡萄球菌的方法。方法利用核酸适配体与其靶标之间的特异性结合能力以及碳纳米管的荧光猝灭特性,构建一种新型快速的金黄色葡萄球菌检测方法。结果该方法特异性良好,与非目标菌株无交叉反应。同时,该方法对金黄色葡萄球菌的检出限为10~5 CFU/mL,线性范围为10~5～10~9 CFU/mL,而且在此浓度范围之间呈现良好的线性关系,线性方程为Y=53.22X-39.99,线性相关系数r~2=0.992。结论该方法具有检测速度快、操作简便、检测成本低等特点,为金黄色葡萄球菌的快速检测提供了一种有效手段。     

金黄色葡萄球菌生物被膜生长动力学分析

利用优化的Liu动力学方程对金黄色葡萄球菌生物被膜的附着过程进行拟合得出:培养温度、培养基p H值、细菌接种浓度都对金黄色葡萄球菌生物被膜的粘附过程有影响,大约在60 min时,金黄色葡萄球菌生物被膜达到粘附平衡。在温度为35℃,p H值为7时,附着的微生物量最大,细菌接种浓度越高,附着的微生物量越大。利用Gompertz模型对生物被膜生长、成熟过程进行拟合得出:金黄色葡萄球菌生物被膜生长曲线呈现典型的S型,具有明显的延滞期、指数期和稳定期;生物被膜生长曲线表现出的S型有别于悬浮菌的生长曲线,其指数期相对较短。金黄色葡萄球菌生物被膜粘附过程、生长和成熟过程的模型矫正拟合度R2(Adj)均在0.97之上。表明Liu动力学方程以及Gompertz模型分别适用于描述外界环境与金黄色葡萄球菌生物被膜附着,生长与成熟的关系。     

金黄色葡萄球菌标准物质的研制

将金黄色葡萄球菌添加到复合保护剂中,经冻干制备成干粉状标准物质,通过比较冻干前后活菌数来验证保护剂保护效果,并研究标准物质的均匀性和稳定性。结果表明复合保护剂的保护率达到51.74%,可以在冻干过程中有效保护菌体,制备的金黄色葡萄球菌标准物质均匀性良好,可以稳定保存半年。该标准物质可用于食品检测实验室质量控制和食品中金黄色葡萄球菌检测结果的评价。     

防腐剂复配对溶血性金黄色葡萄球菌抑菌效果研究

选择不同的防腐剂,通过药敏纸片法、最小抑菌浓度（MIC）法测定金黄色葡萄球菌对其敏感性,利用正交设计筛选最佳防腐剂组合,并对获得的最佳防腐剂组合抑菌效果进行验证。结果表明,金黄色葡萄球菌对乳酸钠、山梨酸钾不敏感,而对乳酸链球菌素（Nisin）、聚赖氨酸、尼泊金乙酯较为敏感,MIC分别为0.005、0.005和0.313mg/mL。正交试验结果表明,当Nisin、聚赖氨酸和尼泊金乙酯浓度分别为0.01、0.02、0.125mg/mL时效果最佳,可使基质中金黄色葡萄球菌降低7.64个数量级。选择这一最佳组合显示,在室温（25℃）里脊肉、牛乳中细菌数在经过4d放置后,仅仅分别增长2.78、1.86个数量级,而猕猴桃汁细菌数降低了0.73个数量级。