The influence of non-lethal temperature on the rate of inactivation of vegetative bacteria in inimical environments may be independent of bacterial species

The influence of non-lethal temperature on the survival of two species of food-borne bacteria under growth-preventing pH and water activity conditions was investigated. Specifically, inactivation rates of four strains of Escherichia coli and three strains of Listeria monocytogenes were determined in culture broth adjusted to pH 3.5 and water activity 0.90, to prevent growth of both species, and for temperatures in the range 5–45 °C at 5 °C intervals. Sixty-three inactivation rates were obtained, plotted on Arrhenius co-ordinates, and lines of best-fit determined by simple linear regression. Differences in the mean inactivation rate of each species at a given temperature were not significant (p < 0.05) with the exception of the rates at 25 °C. The inactivation rate responses of both species were comparable to those reported by McQuestin et al. (Appl. Environ. Microbiol., 75:6963–6972, 2009) for a variety of E. coli strains under a wide range of growth-preventing pH and water activity conditions. The results support the hypothesis that non-lethal temperature is a key factor governing the rate of inactivation of vegetative bacteria in foods when other hurdles prevent their growth and indicate that the temperature effect may also be independent of bacterial species.     

Identification of non-linear microbial inactivation kinetics under dynamic conditions

In this study dynamic microbial inactivation experiments are exploited for performing parameter identification of a non-linear microbial model. For that purpose microbial inactivation data are produced and a differential equation exhibiting a shoulder and a loglinear phase is employed. The derived parameter estimates from this method were used to perform predictions on an independent experimental set at fluctuating temperature. Joint confidence regions and asymptotic confidence intervals of the estimated parameters were compared with previous studies originating from parameter identification under isothermal conditions. The developed approach can provide more reliable estimates for realistic conditions compared to the usual or standard two step approach.     

离心条件对乳酸菌离心存活率的影响

研究了以11%脱脂乳为液体培养基进行培养时,离心力、离心时间和离心温度对乳酸菌在离心过程中损失率、存活率的影响。结果表明:延长离心时间,离心损失率和离心存活率均逐渐降低;在同一离心时间条件下,增大离心力,离心损失率和离心存活率均显著降低(p>0.01);在11%脱脂乳液体培养基中培养的保加利亚乳杆菌、嗜热链球菌,其最适离心条件为:5000r/min离心10min,离心温度为20℃,此时乳酸菌的离心存活率为94.32%。     

耐热乳酸菌热致死动力学模型研究

测定了从云南传统乳制品乳扇中筛选的2种6株耐热乳酸菌在60℃和65℃热处理条件下的菌株存活情况.发现E.faeciumgrx28的耐热能力最强,D60为6.45min,D65为1.32 min与其他菌株有显著差异.分别基于Linear,Weibull和Log-logistic动力学模型,对各耐热乳酸菌在不同热处理条件下的活菌数量的变化情况使用lstopt1.0软件进行数据的曲线拟合及相关系数的分析,确定在60℃的热处理条件下,Weibull模型更适合描述菌株的残活量,而在65℃的热处理条件下,Logistic模型与菌株的存活曲线更为接近,得到各菌株在不同处理条件下的动力学方程.结果表明,乳酸菌在不同热处理条件下,乳酸菌的死亡变化并不相同.     

UV inactivation of bacteria in apple cider

Apple cider, inoculated with Escherichia coli and Listeria innocua, was processed using a simple UV apparatus. The apparatus consisted of a low-pressure mercury lamp surrounded by a coil of UV transparent tubing. Cider was pumped through the tubing at flow rates of 27 to 83 ml/min. The population of E. coli K-12 was reduced by 3.4 +/- 0.3 log after being exposed for 19 s at a treatment temperature of 25 degrees C. The population of L. innocua, which was more resistant to UV, was reduced by 2.5 +/- 0.1 log after being exposed for 58 s. The electrical energy for the process was 34 J/ml and is similar to that for conventional thermal processing. UV processing has the potential to improve the safety and extend the shelf life of apple cider.     

On the use of the Weibull model to describe thermal inactivation of microbial vegetative cells

This paper evaluates the applicability of the Weibull model to describe thermal inactivation of microbial vegetative cells as an alternative for the classical Bigelow model of first-order kinetics; spores are excluded in this article because of the complications arising due to the activation of dormant spores. The Weibull model takes biological variation, with respect to thermal inactivation, into account and is basically a statistical model of distribution of inactivation times. The model used has two parameters, the scale parameter alpha (time) and the dimensionless shape parameter beta. The model conveniently accounts for the frequently observed nonlinearity of semilogarithmic survivor curves, and the classical first-order approach is a special case of the Weibull model. The shape parameter accounts for upward concavity of a survival curve (beta < 1), a linear survival curve (beta = 1), and downward concavity (beta > 1). Although the Weibull model is of an empirical nature, a link can be made with physiological effects. Beta < 1 indicates that the remaining cells have the ability to adapt to the applied stress, whereas beta > 1 indicates that the remaining cells become increasingly damaged. Fifty-five case studies taken from the literature were analyzed to study the temperature dependence of the two parameters. The logarithm of the scale parameter alpha depended linearly on temperature, analogous to the classical D value. However, the temperature dependence of the shape parameter beta was not so clear. In only seven cases, the shape parameter seemed to depend on temperature, in a linear way. In all other cases, no statistically significant (linear) relation with temperature could be found. In 39 cases, the shape parameter beta was larger than 1, and in 14 cases, smaller than 1. Only in two cases was the shape parameter beta = 1 over the temperature range studied, indicating that the classical first-order kinetics approach is the exception rather than the rule. The conclusion is that the Weibull model can be used to model nonlinear survival curves, and may be helpful to pinpoint relevant physiological effects caused by heating. Most importantly, process calculations show that large discrepancies can be found between the classical first-order approach and the Weibull model. This case study suggests that the Weibull model performs much better than the classical inactivation model and can be of much value in modelling thermal inactivation more realistically, and therefore, in improving food safety and quality.     

纳米Fe2O3靶向鸡肉腐败菌光催化抑菌效能 特性研究

目的探索可见光下纳米Fe2O3对鸡肉腐败菌的光催化抑菌效能特性及机制。方法以Escherichia coli及鸡肉腐败菌Pseudomonas fluorescens和Macrococcus caseolyticus为受试菌株,可见光照为激发条件,研究纳米Fe2O3对细菌菌落数量和菌体脂质氧化程度的影响。结果纳米Fe2O3在可见光下能够有效抑制E.coli、P.fluorescens和M.caseolyticus生长繁殖,1.2 mmol/L的H2O2能够增强Fe2O3的光催化抑菌活性;当Fe2O3质量浓度为0.4 g/L时,对P.fluorescens和M.caseolyticus的抑菌作用效果最强;在光催化过程中,P.fluorescens和M.caseolyticu的脂质氧化值随着反应时间延长呈先升高后降低的趋势,在120 min分别达到最大值1.31nmol/mg(细胞干重)和2.14 nmol/mg(细胞干重)。结论可见光条件下,M.caseolyticus对光催化反应比P.fluorescens更加敏感,纳米Fe2O3先引起细菌体内脂质氧化,进而导致菌体细胞破裂而死亡。     

Stability of microencapsulated lactic acid bacteria under acidic and bile juice conditions

The probiotic strains Lactobacillus brevis CCMA1284 and Lactobacillus plantarum CCMA0359 were microencapsulated by spray drying using different matrices – whey powder (W), whey powder with inulin (WI) and whey powder with maltodextrin (WM). Viability of the microencapsulated strains in acid and bile juices and during 90 days of storage (seven and 25 °C) was evaluated. The two strains exhibited high encapsulation efficiency (> 86%) by spray drying. The different matrices maintained L. plantarum viability above six log CFU g⁻¹ at 7 °C for 90 days, whereas similar results for L. brevis were observed only for W. The use of inulin as matrix of encapsulation did not enhance bacterial viability in the evaluated conditions. In general, the use of W and WM as matrices was effective for L. plantarum viability. However, only W was effective for L. brevis in the evaluated conditions. The spray drying technique was successfully adopted for the encapsulation of L. plantarum CCMA0359 and L. brevis CCMA1284 strains.     

High pressure processing combined with selected hurdles: Enhancement in the inactivation of vegetative microorganisms

High pressure processing (HPP) as a nonthermal processing (NTP) technology can ensure microbial safety to some extent without compromising food quality. However, for vegetative microorganisms, the existence of pressure-resistant subpopulations, the revival of sublethal injury (SLI) state cells, and the resuscitation of viable but nonculturable (VBNC) state cells may constitute potential food safety risks and pose challenges for the further development of HPP application. HPP combined with selected hurdles, such as moderately elevated or low temperature, low pH, natural antimicrobials (bacteriocin, lactate, reuterin, endolysin, lactoferrin, lactoperoxidase system, chitosan, essential oils), or other NTP (CO₂, UV-TiO₂ photocatalysis, ultrasound, pulsed electric field, ultrafiltration), have been highlighted as feasible alternatives to enhance microbial inactivation (synergistic or additive effect). These combinations can effectively eliminate the pressure-resistant subpopulation, reduce the population of SLI or VBNC state cells and inhibit their revival or resuscitation. This review provides an updated overview of the microbial inactivation by the combination of HPP and selected hurdles and restructures the possible inactivation mechanisms.     

Pasteurization of milk: the heat inactivation kinetics of milk-borne dairy pathogens under commercial-type conditions of turbulent flow

This is the first study to report kinetic data on the survival of a range of significant milk-borne pathogens under commercial-type pasteurization conditions. The most heat-resistant strain of each of the milk-borne pathogens Staphylococcus aureus, Yersinia enterocolitica, pathogenic Escherichia coli, Cronobacter sakazakii (formerly known as Enterobacter sakazakii), Listeria monocytogenes, and Salmonella was selected to obtain the worst-case scenario in heat inactivation trials using a pilot-plant-scale pasteurizer. Initially, approximately 30 of each species were screened using a submerged coil unit. Then, UHT milk was inoculated with the most heat-resistant pathogens at ~10(7)/mL and heat treated in a pilot-plant-scale pasteurizer under commercial-type conditions of turbulent flow for 15s over a temperature range from 56 to 66°C and at 72°C. Survivors were enumerated on nonselective media chosen for the highest efficiency of plating of heat-damaged bacteria of each of the chosen strains. The mean log(10) reductions and temperatures of inactivation of the 6 pathogens during a 15-s treatment were Staph. aureus >6.7 at 66.5°C, Y. enterocolitica >6.8 at 62.5°C, pathogenic E. coli >6.8 at 65°C, C. sakazakii >6.7 at 67.5°C, L. monocytogenes >6.9 at 65.5°C, and Salmonella ser. Typhimurium >6.9 at 61.5°C. The kinetic data from these experiments will be used by the New Zealand Ministry of Agriculture and Forestry to populate the quantitative risk assessment model being developed to investigate the risks to New Zealand consumers from pasteurized, compared with nonpasteurized, milk and milk products.     

Lytic and nonlytic mechanism of inactivation of gram-positive bacteria by lysozyme under atmospheric and high hydrostatic pressure

A different behavior was observed in three gram-positive bacteria exposed to hen egg white lysozyme by plate counts and phase-contrast microscopy. The inactivation of Lactobacillus johnsonii was accompanied by spheroplast formation, which is an indication of peptidoglycan hydrolysis. Staphylococcus aureus was resistant to lysozyme and showed no signs of peptidoglycan hydrolysis, and Listeria innocua was inactivated and showed indications of cell leakage but not of peptidoglycan hydrolysis. Under high hydrostatic pressure, S. aureus also became sensitive to lysozyme but did not form spheroplasts and was not lysed. These results suggested the existence of a nonlytic mechanism of bactericidal action of lysozyme on the latter two bacteria, and this mechanism was further studied in L. innocua. Elimination of the enzymic activity of lysozyme by heat denaturation or reduction with beta-mercaptoethanol eliminated this bactericidal mechanism. By means of a LIVE/DEAD viability stain based on a membrane-impermeant fluorescent dye, the nonlytic mechanism was shown to involve membrane perturbation. In the absence of lysozyme, high-pressure treatment was shown to induce autolytic activity in S. aureus and L. innocua.     

不同粉碎条件对葡萄籽超微粉破壁率的影响

以赤霞珠葡萄籽为原料,研究不同的冷冻温度和粉碎时间对葡萄籽超微粉细胞破壁率的影响。借鉴理想破碎模型单元破碎率计算法,测定葡萄籽的细胞直径以及葡萄籽超微粉的粒径,并统计粒径的分布概率研究破壁率。结果表明:同一冷冻温度下,除个别条件下存在例外,葡萄籽细胞破壁率随着粉碎时间的延长有明显的升高趋势,且在-15和-25℃时破壁率与粉碎时间具有线性关系;在同一粉碎时间下,随着温度的降低破壁率有升高的趋势但不呈线性,在粉碎时间延长至75min时破壁率已达到较高水平,温度对其影响微弱。超微粉粒径小于葡萄籽细胞平均直径时,破壁率为100%;超微粉粒径大于葡萄籽细胞平均直径时,破壁率随着粒径的增大而减小。推荐较好的葡萄籽超微粉破碎条件为:冷冻温度-15℃、粉碎时间75min。     

Inactivation model equations and their associated parameter values obtained under static acid stress conditions cannot be used directly for predicting inactivation under dynamic conditions

Organic acids (e.g., lactic acid, acetic acid and citric acid) are popular preservatives. In this study, the Listeria innocua inactivation is investigated under dynamic conditions of pH and undissociated lactic acid ([LaH]). A combined primary (Weibull-type) and secondary model developed for the L. innocua inactivation under static conditions [Janssen, M., Geeraerd, A.H., Cappuyns, A., Garcia-Gonzalez, L., Schockaert, G., Van Houteghem, N., Vereecken, K.M., Debevere, J., Devlieghere, F., Van Impe, J.F., 2007. Individual and combined effects of pH and lactic acid concentration on L. innocua inactivation: development of a predictive model and assessment of experimental variability. Applied and Environmental Microbiology 73(5), 1601-1611] was applied to predict the microbial inactivation under dynamic conditions. Because of its non-autonomous character, two approaches were proposed for the application of the Weibull-type model to dynamic conditions. The results quantitatively indicated that the L. innocua cell population was able to develop an induced acid stress resistance under dynamic conditions of pH and [LaH]. From a modeling point of view, it needs to be stressed that (i) inactivation model equations and associated parameter values, derived under static conditions, may not be suitable for use as such under dynamic conditions, and (ii) non-autonomous dynamic models reveal additional technical intricacies in comparison with autonomous models.     

不同粉碎条件对葡萄籽超微粉破壁率的影响

以赤霞珠葡萄籽为原料,研究不同的冷冻温度和粉碎时间对葡萄籽超微粉细胞破壁率的影响。借鉴理想破碎模型单元破碎率计算法,测定葡萄籽的细胞直径以及葡萄籽超微粉的粒径,并统计粒径的分布概率研究破壁率。结果表明:同一冷冻温度下,除个别条件下存在例外,葡萄籽细胞破壁率随着粉碎时间的延长有明显的升高趋势,且在-15和-25℃时破壁率与粉碎时间具有线性关系;在同一粉碎时间下,随着温度的降低破壁率有升高的趋势但不呈线性,在粉碎时间延长至75min时破壁率已达到较高水平,温度对其影响微弱。超微粉粒径小于葡萄籽细胞平均直径时,破壁率为100%;超微粉粒径大于葡萄籽细胞平均直径时,破壁率随着粒径的增大而减小。推荐较好的葡萄籽超微粉破碎条件为:冷冻温度-15℃、粉碎时间75min。      

Time to growth and inactivation of three STEC outbreak strains under conditions relevant for fermented sausages

Published models predicting growth and survival capabilities of shiga-toxin-producing Escherichia coli (STEC) under a_w and lactic acid stress were validated by performing experiments with fermented sausage associated outbreak strains. Strain variation in inactivation and time to growth (TTG) were investigated for strains representing three serotypes (O103, O111, and O157). The TTG and growth boundaries of each strain were compared with predictions of a model for generic acid adapted E. coli and survival with predictions of two inactivation models. In addition, the influence of strain variation on the performance of the inactivation models, in terms of bias and accuracy factors, was illustrated. Strains with induced acid tolerance were used in broths containing 50 or 110 mM total lactic acid. The concentration of undissociated lactic acid (HLac) was adjusted by setting the pH-value, and water activity (0.900 to 0.995 depending on experiment) was adjusted by adding NaCl. The survival capabilities of the outbreak strains were good compared to the model predictions. The average bias factors of inactivation model predictions were within a factor of 2.2 depending on the strain used to validate the model indicating that inactivation rates of outbreak strains were slower than predicted. However, the observed rates were similar to the rates of a previously studied acid tolerant generic E. coli strain. Similarly, the time to growth of two of the strains (O103 and O157) was comparable with model predictions, whereas the growth capability of the third strain (O111) was lower than predicted. These results suggest that the properties of the most tolerant sausage outbreak strains are comparable to tolerant generic E. coli strains, which imply that suitable non-pathogenic E. coli strains are valid surrogates for fermented sausage outbreak strains. The relative sensitivity of strains depended on the environmental parameters and the response evaluated. The strain with the smallest log reduction at 20 °C was O157, whereas it was strain O103 at 8 °C. Under conditions unfavorable for growth, the time to growth was much shorter for strains O103 and O157 than for strain O111, whereas differences between strains were negligible under conditions favorable for growth. Depending on the response variable and the specific application the limitation of not addressing strain variation may lead to biased, fail-dangerous, predictions. Thus, solutions on how to best address strain variation in the development and validation of predictive models are needed.     

Pulsed electric fields inactivation of attached and free-living Escherichia coli and Listeria innocua under several conditions

The use of pulsed electric fields (PEF) is considered as a mild process in the inactivation of microorganisms present in liquid food products. PEF treatments of Escherichia coli and Listeria innocua suspended in milk and phosphate buffer, with same pH and same conductivities, yielded to similar inactivation. Reduction rates obtained in distilled water indicated that conductivity of the food product is a main parameter in bacterial inactivation. Bacteria attached to polystyrene beads were inactivated by PEF at a greater (E. coli) or equal rate (L. innocua) than free-living bacteria. Base on the use of selective and non-selective enumeration media, no clear indications were obtained for sublethal damage of microorganisms surviving the PEF treatment. E. coli cells subjected to 60 pulses at 41 kV/cm were examined by transmission and scanning electron microscopy. Changes in the cytoplasm were observed and the cell surface appeared rough. The cells outer membranes were partially destroyed allowing leaking of cell cytoplasm.     

Menadione-catalyzed luminol chemiluminescence assay for the rapid detection of viable bacteria in foods under aerobic conditions

A menadione-catalyzed luminol chemiluminescence assay was developed for the rapid detection and estimation of viable bacteria in foods. The principle of this assay is based on the extracellular menadione-catalyzed active oxygen spieces (O2- and H2O2) generated by the activity of NAD(P)H:menadione oxidoreductase in viable cells. This luminol chemiluminescence assay requires 10 min for the incubation of cells with menadione and then 2 s for the measurement of chemiluminescence intensity after an injection of luminol solution without the treatment of cell lysis. This method was evaluated using liquid food samples of milk, vegetable juice, green tea, and coffee spiked with Escherichia coli ATCC 25922. The study result revealed that E. coli contamination at 1 to 10 CFU/ml in these foods could be detected after incubation at 37 degrees C for 7 h in an enrichment medium; however, the green tea and coffee samples requires filtration. This method could be a useful tool for the rapid evaluation of microbial food contamination.     

提高棉织物低带液量下活性染料的固色率

活性染料在低带液量条件下具有较高的固色率,为进一步了解低带液量时影响活性染料固色率的因素,试验探讨了棉织物带液量、Na₂CO₃质量浓度、固色温度、固色时间、Na₂SO₄加入量和加入方式、染料上染量、染料直接性对固色率的影响。结果表明:棉织物在带液量为25%～35%时,染料的固色率最高;碱剂量、固色时间、固色温度、上染量对染料水解影响较小。在低带液量下,染料在纤维上的分布状态与染料结构是影响固色率的主要因素,进入纤维内部的染料可实现内部扩散和固色,较难发生水解。染料直接性越大,活性基团活性越高且数量越多,染料固色率就越高。     

The influence of the temperature increase rate on the accuracy of diffusion parameters estimated under non-isothermal conditions

The accuracy of the diffusion parameters estimated under non-isothermal conditions was studied using computer-generated pseudo-experimental points. The data sets were obtained from Fick's 2nd law and an Arrhenius-type temperature dependence of the diffusivity. A normally distributed error was randomly attributed to the model data. The pseudo-experimental points thus obtained were then fitted to the model equation, with non-linear regression using the Simplex algorithm, yielding the estimates of the pre-exponential factor and activation energy. Several linear temperature profiles were studied, from 0.014 to 16°C min⁻¹. It was concluded that the accuracy of the parameters estimation depends on the temperature increase rate. The effects of the pre-exponential factor, the activation energy, the range of temperature and the magnitude of the experimental error on the value of the temperature increase rate that led to maximum accuracy of the parameters estimates were also studied.