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.     

二氧化氯对苹果表面Listeria monocytogenes杀菌效果的研究

利用A PI Listeria鉴定系统对从榨汁苹果表面分离的Listeria monocytogenes疑似菌落进行了鉴定,并将原料清洗与杀菌过程相结合,通过4因素中心组合试验设计(CCD),以杀菌效果(以杀菌效率的负对数表示,即logNN )为响应值,研究二氧化氯溶液浓度、杀菌温度、处理时间和杀菌液pH 等因素对杀 0菌效果的影响,用数学方法描述二氧化氯对苹果表面Listeria m onocytogenes的杀菌规律。结果表明:二氧化氯浓度对杀菌效果影响最大,杀菌液pH 次之,杀菌温度影响最小,延长杀菌时间对杀菌效果的影响不显著,二氧化氯溶液浓度、杀菌温度和pH 之间对杀菌效果有协同作用,建立的数学模型其预报结果与实测结果拟合基本一致,相对误差在 5.0% 以内。 ±     

Effect of water activity on inactivation of Listeria monocytogenes and lactate dehydrogenase during high pressure processing

The aim of this study was to investigate the effect of water activity (a_w) on the inactivation of Listeria monocytogenes and lactate dehydrogenase (LDH) during high pressure processing (HPP). For microbial inactivation lyophilized cells of L. monocytogenes 19,115 were left dry or were suspended in 10 ml of 0.1% peptone water, 10 ml of glycerol, or mixtures of glycerol and peptone water. All samples of various a_ws were high pressure (HP) processed at ambient temperature at 600 MPa for 300 s. Following HPP, samples were serially diluted in 0.1% peptone and spread-plated on Tryptic Soy agar supplemented with Yeast Extract. For enzyme inactivation, 4.2 mg of lyophilized LDH was suspended in 2 ml of 100 mM phosphate buffer (pH 7.4), 2 ml of peptone water or glycerol, or in 2 ml mixtures of glycerol and peptone water. A lyophilized sample with no added liquid was also included. All enzyme samples were subjected to HPP as described above. After HPP, LDH was diluted to 0.28 μg/ml in 100 mM phosphate buffer (pH 7.4). LDH activity was assessed by measuring the change in concentration of β-NADH as a function of time. Dynamic light scattering analysis (DLS) was performed to examine the size distribution, polydispersity, and hydrodynamic radius of LDH before and after HPP. No significant difference in CFU/g was observed between lyophilized cells not subjected to HPP and lyophilized cells subjected to 600 MPa for 300 s (P < 0.05). However, lyophilized cells that were suspended in 100% to 60% peptone water showed a ~ 7.5-log₁₀ reduction when subjected to HPP. Survival of L. monocytogenes following HPP significantly increased (P < 0.05) when the peptone water concentration was decreased below 60% (a_w ~ 0.8). DLS results revealed that LDH suspended in buffer underwent aggregation following HPP (600 MPa, 300 s). Inactivation rate constants obtained using a first-order kinetic model indicated that untreated and HP processed lyophilized LDH had similar activities. When LDH was subject to HPP in solutions containing glycerol, enzyme activity decreased as the water content increased (r² = 0.95). Lyophilization completely protected L. monocytogenes and LDH from inactivation by high pressure. Furthermore, enzyme activity and cell survival increased as water activity was decreased. We postulate low a_w results in protein stabilization, which prevents protein denaturation and cell death during HPP.     

肉桂醛对猪肉糜中单核细胞增生李斯特菌的 抑制作用及其机制

目的研究不同温度条件下肉桂醛对猪肉糜中单核细胞增生李斯特菌的抑制作用,并分析其机制。方法在不同温度(55、60、65和70℃)条件下,对添加不同浓度肉桂醛的猪肉糜中单核细胞增生李斯特菌的活细胞变化曲线进行测定分析,并采用扫描电子显微镜技术研究肉桂醛对其菌体形态的影响。结果在猪肉糜中添加肉桂醛能明显增大单核细胞增生李斯特菌的热失活速率,缩短热处理的时间,这种促进作用也随着肉桂醛浓度的增大而逐渐加强。在70℃下,添加有1.0%肉桂醛的猪肉糜样品在0.5 min之内可使得单核细胞增生李斯特菌活细胞的数目从9.03 Log CFU/g迅速减少到2.89 Log CFU/g,而未添加肉桂醛的对照样品则需要3 min。扫描电子显微镜结果显示,肉桂醛处理后单核细胞增生李斯特菌的菌体变长,细胞色泽暗沉。结论肉桂醛能够显著改变单核细胞增生李斯特菌的形态,降低其在猪肉糜中的热抵抗能力,因而加快了其热失活速率。本研究为肉桂醛在食品中食源性致病菌的安全控制领域的应用提供了参考。     

Effect of preinoculation growth media and fat levels on thermal inactivation of a serotype 4b strain of Listeria monocytogenes in frankfurter slurries

Preinoculation growth conditions and fat levels were evaluated for effects on the heat resistance of Listeria monocytogenes strain MFS 102 in formulated frankfurter slurries and on frankfurter surfaces. Comparison of linear inactivation rates (D-values) for cells heated in frankfurter slurry showed that growth conditions were significant (P<0.05) factors affecting subsequent thermal resistance. The average D(60 degrees C)-values for the five preinoculation growth media tested from most resistant to least heat resistant were: tryptic soy broth with 0.6% yeast extract (TSBYE) (2.2 min) and 8.5% fat slurry (2.2 min), followed by 23% fat slurry (1.7 min) and 11% fat slurry (1.7 min), and then TSYBE with quaternary ammonium compounds added (TSBYE+Q) (1 min). The fat level in the frankfurter heating media also had a significant (P<0.05) effect on the thermal death rate of L. monocytogenes. Cells heated in 8.5% fat slurry had a significantly higher (P<0.05) D(60 degrees C)-value (2.2 min) than those heated in 11% fat (1.0 min) and 23% fat slurry (0.9 min). Growth media (TSBYE, 8.5% fat slurry, and TSBYE+Q), and fat level (15% and 20%), however, were not significant factors (P>0.05) affecting thermal inactivation rates on frankfurter surfaces. Heat inactivation rates were consistently higher on frankfurter surfaces compared to similar treatments done in frankfurter slurry. On frankfurter surfaces, a 2.3- to 5.1-log(10) reduction was achieved after 15 min depending on frankfurter surface type. The time necessary to achieve a 3-log(10) reduction using post-processing pasteurization of frankfurters in a hot water-bath at 60 degrees C almost doubled for cells grown in TSBYE and heated in 23% fat frankfurter slurry (19.6 min) versus cells grown and heated in 8.5% fat frankfurter slurry (10.8 min).     

鲜切结球莴苣中单增李斯特菌生长预测模型的建立

为建立不同温度下鲜切结球莴苣中单增李斯特菌生长模型,将单增李斯特菌接种到鲜切结球莴苣表面,并于不同温度下贮藏,获得其在4、8、16、24和32℃下的生长数据,选用Gompertz模型进行拟合,建立初级生长模型。在此基础上建立二级模型研究温度对初级模型中单增李斯特菌生长动力学参数的影响,并进行数学检验。结果表明,对最大比生长速率和延滞时间建立平方根模型,结果呈良好的线性关系,相关系数R2分别为0.977 2和0.984 7,所建立的预测模型能很好地描述不同温度下单增李斯特菌的生长动态。     

Effects of ultra high hydrostatic pressure on Listeria monocytogenes and natural flora in broth, milk and fruit juices

Listeria monocytogenes was subjected to ultra high hydrostatic pressure (UHHP) treatments from 200 to 700 MPa at 25 °C in broth, raw milk, peach juice and orange juice. Survivor curves showed that cell death increased as pressure increased. After 10 min pressure treatment at 400 MPa reductions of about 2.09 and 2.76 log CFU mL^?1 in aerobic bacteria and L. monocytogenes, respectively, were produced in raw milk, this increased to 5.09 and 6.47 log CFU mL^?1, respectively, at 600 MPa. Death of bacteria at UHHP treatment was greater in orange juice than peach juice, and in peach juice than milk. Listeria monocytogenes was more sensitive to increased pressure than increased pressurization time. Injury of L. monocytogenes occurred from 0 to 100%. Factors effecting the rate of microbial inactivation are: pressure, age of cell, composition of medium, and pressurization time. UHHP inactivation can be used to extend shelf life and increase food quality during storage, and may also contribute to inactivation of L. monocytogenes.     

酸性电解水对副溶血性弧菌和单核细胞增生李斯特氏菌杀菌效果的比较研究

目的研究不同浓度酸性电解水对副溶血性弧菌和单核细胞增生李斯特氏菌单增李斯特菌的杀菌作用,比较酸性电解水对革兰氏阴性菌和革兰氏阳性菌的杀菌效果。方法采用传统的平板计数法进行细菌计数,扫描电镜观察细菌细胞的形态变化,琼脂糖胶电泳检测细菌DNA变化和BSA法进行细菌蛋白质泄露测定。结果酸性电解水对副溶血性弧菌和单核细胞增生李斯特氏菌均具有很强的杀灭效果,其杀菌效果随着电解水浓度的增加而增强。较高浓度的电解水处理后,副溶血性弧菌几乎全部杀灭,单核细胞增生李斯特氏菌菌落总数降低了1.45 log CFU/mL。扫描电镜实验表明,经酸性电解水处理的副溶血性弧菌坍塌明显,且随着酸性电解水浓度的升高,细胞形态崩解严重,细菌形状模糊。结论相较于单核细胞增生李斯特氏菌,酸性电解水对革兰氏阴性菌副溶血性弧菌有明显的杀菌效果     

Searching for new mathematical growth model approaches for Listeria monocytogenes

ABSTRACT:  Different secondary modeling approaches for the estimation of Listeria monocytogenes growth rate as a function of temperature (4 to 30 °C), citric acid (0% to 0.4% w/v), and ascorbic acid (0% to 0.4% w/v) are presented. Response surface (RS) and square-root (SR) models are proposed together with different artificial neural networks (ANN) based on product functions units (PU), sigmoidal functions units (SU), and a novel approach based on the use of hybrid functions units (PSU), which results from a combination of PU and SU. In this study, a significantly better goodness-of-fit was obtained in the case of the ANN models presented, reflected by the lower SEP values obtained (< 24.23 for both training and generalization datasets). Among these models, the SU model provided the best generalization capacity, displaying lower RMSE and SEP values, with fewer parameters compared to the PU and PSU models. The bias factor (B_f) and accuracy factor (A_f) of the mathematical validation dataset were above 1 in all cases, providing fail-safe predictions. The balance between generalization properties and the ease of use is the main consideration when applying secondary modeling approaches to achieve accurate predictions about the behavior of microorganisms.     

单核细胞增生李斯特菌的毒力因子

目的单核细胞增生李斯特菌(Listeria monocytogenes,Lm)属于革兰阳性无芽孢杆菌李斯特菌属,主要通过食物传播。Lm的致病性与毒力因子密切相关,研究其毒力因子对认识致病机理有着重要意义。方法对Lm重要的毒力因子(溶血素、磷脂酶、内化素、肌动蛋白、P60蛋白等)进行综述。结果溶血素是一个多功能的毒力因子,对于Lm逃离吞噬细胞囊是必需的。Lm能产生两种磷脂酶C:磷脂酰肌醇磷脂酶C和磷脂酰胆碱磷脂酶C,协助细菌的细胞内复制。肌动蛋白使得Lm在宿主细胞间能够扩散。内化素与Lm的侵袭力有关。P60蛋白是Lm的主要免疫原性抗原。结论Lm毒力因子研究的深入对李斯特菌病的防治将带来深远的影响。     

D and z Values of Salmonella, Listeria innocua, and Listeria monocytogenes in Fully Cooked Poultry Products

: The D and z values of Salmonella, Listeria innocua, and Listeria monocytogenes were obtained for different ready‐to‐eat poultry products, including chicken, turkey, and duck. The D values of Salmonella, L. innocua, and L. monocytogenes were 151.5 to 0.1 min at 55 to 70°C, and the z values of Salmonella, L. innocua, and L. monocytogenes were 4.9 to 7.0 °C. Significant differences were found for the heat resistance of Salmonella, L. innocua, and L. monocytogenes among turkey, duck, and chicken products, indicating that the kinetic values of a certain pathogen in a specific product should be used for determining process lethality in fully cooked and vacuum‐packaged poultry products during post‐cook heat treatments.     

单核细胞增生李斯特氏菌的PCR检测方法

研究比较了裂解法、热煮沸法、试验盒法提取单增李斯特氏菌DNA的效果 ,认为Promega试剂盒法提取的DNA ,得率高、纯度好。同时根据单增李斯特氏菌的毒力相关基因的 5对引物 ,进行了引物的特异性研究 ,结果发现inlA引物、inlB引物、plcA引物特异性好 ,可以用于食品中单增李斯特氏菌的检测     

Antimicrobial activity of Ginkgo biloba leaf extract on Listeria monocytogenes

ABSTRACT: The antimicrobial activities of Ginkgo biloba leaf extract (GBE) and the combined effects of GBE and sodium EDTA (sodium Ethylenediaminetetraacetic acid) against Listeria monocytogenes were determined at 4 °C, 25 °C, and 37 °C. Listeria monocytogenes grown at 37 °C for 24 h was inoculated (6 to 7 log CFU/mL) into BHI broth containing either GBE or GBE and EDTA (1.6 mg/mL) with various GBE concentrations of 0.1, 0.25, 0.5, 1, 2.5, 5.0, 7.5, 10.0, 15.0, or 20.0% vol/vol and stored at 4 °C, 25 °C, and 37 °C. The inhibitory effect of the GBE was more pronounced at low temperature of 4 °C. GBE was effective in inhibiting microbial growth. Addition of EDTA enhanced antimicrobial activity of GBE.     

一株食源性多重耐药单核细胞增生李斯特菌的耐药性研究

本研究对食品样本中分离的一株多重耐药单核细胞增生李斯特菌进行耐药机制的探讨,以期对食源性单核细胞增生李斯特菌多重耐药现象的控制提供理论依据。本文通过聚合酶链式反应筛选耐药决定因子,质粒消除及自然转化实验对耐药决定因子进行定位及传播能力的探讨,最后通过传代实验验证该菌株多重耐药性传播的稳定性。结果表明,对检测到的多重耐药菌株LM78(耐受氯霉素、红霉素、链霉素、四环素、复方新诺明)进行相关耐药基因检测,检测到cat、erm B、tet S 3个耐药基因。质粒消除后MIC值下降到敏感范围,且该质粒可通过自然转化在不同菌属间传递,说明这些耐药基因存在于质粒上。该质粒在无抗生素选择压力下连续传代,仍具有较高稳定性。食源性致病菌多重耐药性有可能通过不同细菌种属间转移,进而由食物链向人类传播,对人类健康造成潜在的威胁。     

Effects and interactions of sodium lactate, sodium diacetate, and pediocin on the thermal inactivation of starved Listeria monocytogenes on bologna

The effects and interactions of temperature (56.3-60 °C), sodium lactate (SL; 0-4.8%), sodium diacetate (SDA; 0-2.5%), and pediocin (0-10,000 AU) on starved Listeria monocytogenes (10⁷ CFU/g) on bologna were investigated. Bologna slices containing SL and SDA in the formulation were dipped in pediocin, surface inoculated, and treated at various temperatures using combinations of parameters determined by central composite design. D-values were calculated. The observed D-values ranged from 2.8 min at 60 °C to 24.61 min at 56.3 °C. Injury ranged from 9.1 to 76% under various conditions. The observed D-values were analyzed using second order response surface regression for temperature, SL, SDA, and pediocin, and a predictive model was developed. Predicted D-values were calculated and ranged from 3.7 to 19 min for various combinations of parameters. Temperature alone reduced the predicted D-values from 33.96 min at 56.3 °C to 11.51 min at 60 °C. Addition of SL showed a protective effect. Other combination treatments either reduced or increased D-values depending on temperature. The combination of SL and SDA was effective at lower temperatures, however, higher levels of SDA at higher temperatures made the organism more heat resistant. Pediocin (up to 5000 AU) with increasing temperature and SDA reduced D-values. Depending on temperature and concentration, the interactions between various additives can affect thermal inactivation of L. monocytogenes on bologna. Starvation rendered L. monocytogenes more susceptible to heat and additives.     

单核细胞增生性李斯特菌自溶素研究进展

自溶素是由细菌产生的可以降解细菌细胞壁的肽聚糖水解酶,参与细菌的分裂、细胞壁更新、细菌自溶等生理过程。自溶素也是细菌的重要毒力因子之一,参与细菌的黏附与侵袭,与细菌的致病性密切相关。本文对单核细胞增生性李斯特菌的胞壁水解酶p60、酰胺酶Ami、自溶素IspC、自溶素Auto与胞壁水解酶MurA等自溶素的蛋白结构、生理功能及致病性进行了简要介绍。     

Control of Listeria monocytogenes in Vacuum-Packaged Pre-Peeled Potatoes

Growth of Listeria monocytogenes on the surface of fresh peeled potatoes, treated with sulfite or a commercial browning inhibitor (CBI), packaged under vacuum and stored at 4,15 and 28°C was determined. At 4°C, L. monocytogenes did not grow in all treated potatoes even after 21 days. At 15°C, L. monocytogenes grew to 7 log₁₀ CFU/g within 12 days in the potatoes treated with sulfite or CBI. At 28°C, L. monocytogenes population was greater than 3 log₁₀ CFU/g by 24 h in all samples regardless of treatment. Sulfites or a CBI appeared to provide a measure of safety in pre-peeled potatoes packaged under vacuum when kept at proper refrigeration temperatures.     

Recent developments in molecular sub-typing of Listeria monocytogenes

As a vast majority of the human listeriosis cases are caused by serotypes 1/2a, 1/2b and 4b strains, it is imperative that strains from clinical as well as from food and environment are further characterised so that accurate and timely epidemiological determination of sources of the contamination can be established to minimise the disease burden. Recent developments in the field of genomics provide a great opportunity to use these tools towards the development of molecular sub-typing techniques with a greater degree of discrimination spanning the entire length of the genome. This brief review summarises a few of these DNA-based techniques with an emphasis on DNA microarray and other whole genome sequencing-based approaches and their usefulness in Listeria monocytogenes sub-typing and outbreak investigations.     

乳酸菌细菌素Durancin GL对单增李斯特菌的抗菌活性及机制

单增李斯特菌广泛分布于肉类、禽类、蛋类、乳制品及蔬菜中,且适应能力强,即使在4 ℃的冷藏环境下仍可生长繁殖,是食品中主要的食源性致病菌之一。乳酸菌细菌素Durancin GL是由干酪中肠球菌产生的一种新型细菌素,对单增李斯特菌具有靶向抑制作用。本实验研究了Durancin GL对单增李斯特菌的抗菌活性及作用机制。通过最小抑菌浓度和抑菌动力学实验检测Durancin GL对单增李斯特菌的抑制作用,结合监测胞内物质泄漏、菌体存活情况以及形态学分析,探讨Durancin GL对单增李斯特菌的抑菌机制。Durancin GL对单增李斯特菌最小抑菌浓度为（2.5±0.4）mg/L,可引起李斯特菌细胞质泄漏,增加细胞外液电导率,导致菌体细胞死亡,从而发挥其抑菌活性。