Staphylococcus aureus Growth and Enterotoxin Production in Mushrooms

An enterotoxin A (SEA) producing strain of Staphylococcus aureus was inoculated onto mushrooms and incubated under simulated pre-and post-canning conditions. S. aureus grew and produced SEA in mushrooms incubated at 25°C in 10% to 20% NaCl. Growth and SEA production occurred when mushrooms were stored in plastic bags at 37°C, but not at 25°C. Mushrooms heat-processed at 121.1°C supported S. aureus growth and SEA production. No SEA was recovered from mushrooms inoculated with 1 or 10 ng SEA/g and heated at 121.1°C for 4.5 min. At higher inoculated SEA levels (100 and 1,000 ng/g), SEA was detectable after 10 min of heating. Staphylococcal enterotoxin could be present in canned mushrooms as a result of pre-, but more likely post-processing contamination.     

Interrelationship Between Water Activity, Temperature and 5.5 Percent Oxygen on Growth and Enterotoxin A Secretion by Staphylococcus aureus in Precooked Bacon

The limiting water activity (a_w) for the growth of S. aureus A-100 sealed in cans at an oxygen concentration of 5.5% was 0.87 at 37°C and 0.91% at 20°C, values intermediate to those obtained previously for aerobic and anaerobic storage. Maximum populations attained at 37°C and 20°C were equal and once achieved population levels declined more slowly at 20°C than at 37°C. Enterotoxin A production was detected for population levels exceeding 10⁶ CFU/g bacon and were 100 ng/g bacon at 37°C and 16 ng/g bacon at 20°C. It is suggested that while temperature and oxygen are important considerations, a_w, being the main parameter in controlling microbial growth, should be measured directly rather than depending on indirect measurements such as the moisture-salt ratio.     

温度和接种量对搅打奶油中金黄色葡萄球菌生长及产肠毒素的影响

目的：研究金黄色葡萄球菌在搅打奶油中的生长以及产毒特性。方法：将不同浓度的初始接种量的产A型肠毒素金黄色葡萄球菌菌液接种到奶油中并搅打成型,低接种量控制在2～3（lg（CFU/g））,高接种量控制在4（lg（CFU/g））以上。定时测量不同贮存温度条件下奶油中的金黄色葡萄球菌的菌落总数以及产毒状况。结果：36 ℃条件下生长速率最快,其他温度条件下依次降低;低初接种量水平下,只有36 ℃条件下于27 h检测到产生毒素,其他温度条件下未检测到毒素产生;高初接种量水平条件下,在36 ℃于12 h、25 ℃于24 h、15 ℃于66 h检测到产生毒素,5 ℃条件下金黄色葡萄球菌既不生长,又不产毒。结论：随着温度的升高,金黄色葡萄球菌的生长速率随之升高,产肠毒素的时间也随之缩短,高初始接种量水平肠毒素产生的时间短于低初始接种量水平,肠毒素的产生时间为一般在对数期的中后期,此时金黄色葡萄球菌菌落总数≥6（lg（CFU/g））。     

Growth and Production of Enterotoxin A by Staphylococcus aureus on "Home-style" French Fries

Staphylococcus aureus strains were inoculated onto fresh-cut oil-blanched potato strips stored at 21 or 26.7 °C for up to 9 h to determine if the microorganism was capable of growth and staphylococcal enterotoxin (SE) production. Potato strips were assayed for SE using a commercial ELISA kit prior to and following finish frying. S. aureus increased by 2.5 to 2.8 log CFU/g over 9 h at 26.7 °C, and SE was detected after 5 h. SE remained serologically detectable following finish frying of the potato strips. It is recommended that oil-blanched potato strips stored at 26.7 °C be finish fried and served, or discarded, within 3 to 4 h to prevent possible production of SE.     

On the Minimal Water Activity for Growth of Staphylococcus aureus

S. aureus strains ATCC 6538 P, C-243, S-6, and 196 E were unable to grow at 35°C in brain heart infusion, pork extract or beef extract media in which the water activity was adjusted to 0.837—0.839 with sodium chloride. These results are different from some literature reports which indicated that S. aureus could grow at a_w 0.83—0.84. Possible reasons for the differences in the minimal a_w reported to be required for growth of S. aureus are discussed.     

Effect of Water Activity on the Growth Kinetics of Staphylococcus aureus in Ground Bread Crumb

The kinetics of Staphylococcus aureus (strains A, B, D) growth in bread crumb were determined as a function of water activity (A_w) from 0.836 to 0.909 at pH 5.2 to 5.5 and at 35°c. Adding glycerol to the dough or equilibrating the bread over saturated salt solutions adjusted the A_w of the bread. Growth kinetics data, plotted as enumerated colony counts versus incubation time, were fitted using the logistic function to determine maximum growth rates. Similar maximum growth rates resulted, irrespective of the method used to adjust A_w. Extrapolation of growth rate‐A_w results predicts the A_w corresponding to a zero growth rate.     

Growth and Enterotoxin A Production by Staphylococcus aureus in Precooked Bacon in the Intermediate Moisture Range

Different water activities were obtained in precooked bacon by varying the frying time. Water activity (a_w) correlated best to the moisture, salt and protein content. When stored aerobically at 37°C, S. aureus A100 was capable of rapid growth in precooked bacon at a a_w of 0.84 or above, whereas at 20°C a a_w of 0.88 or higher was required. Under anaerobic storage at 37°C, growth was observed at a a_w of 0.90, and at 20°C slight growth was noted at a a_w of 0.91. The increase in the minimal a_w required for aerobic growth at the lower temperature was reflected in the differences between the isotherms obtained at 37°C and 20°C. The maximum populations achieved were higher for samples stored aerobically. Enterotoxin A (19–821 ng/g) was found in all aerobically stored samples where growth occurred. Enterotoxin A (38–109 ng/g) was also found in all anaerobically incubated samples where the population of S. aureus increased more than one logarithmic cycle.     

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。     

Combined Effects of Temperature,pH and Water Activity on Predictive Ability of Microbial Kinetic Inactivation Model

It is well known that temperature is the key factor controlling the microbial survival/inactivation. However, the interactive effects of further stressing environmental conditions may influence microbial behaviour. The objective of this work was to include, in the inactivation model, temperature, pH and a_w effects using a black box polynomial model, aiming at accurate prediction. Data of Listeria innocua obtained within the temperature range of 52.5 and 65.0 °C, pH of 4.5, 6.0 and 7.5, and a_w of 0.95 and 0.99 were used for model assessment. The relations of such parameters with temperature, a_w and pH were assumed to be polynomials.     

Effects of Frozen Storage on Survival of Staphylococcus aureus and Enterotoxin Production in Precooked Tuna Meat

This study investigated the survival of Staphylococcus aureus in precooked tuna meat for producing canned products during frozen storage (?20 ± 2 °C) as well as its growth and enterotoxin production at 35 to 37 °C after the storage. Samples (50 ± 5 g) of precooked albacore (loin, chunk, and flake) and skipjack (chunk and flake) tuna were inoculated with 5 enterotoxin‐producing strains of S. aureus at a level of approximately 3.5 log CFU/g and individually packed in a vacuum bag after 3 h incubation at 35 to 37 °C. Vacuum‐packed samples were stored in a freezer (?20 ± 2 °C) for 4 wk. The frozen samples were then thawed in 37 °C circulating water for 2 h and incubated at 35 to 37 °C for 22 h. Populations of S. aureus in all precooked tuna samples decreased slightly (<0.7 log CFU/g) after 4 wk of storage at ?20 ± 2 °C, but increased rapidly once the samples were thawed and held at 35 to 37 °C. Total S. aureus counts in albacore and skipjack samples increased by greater than 3 log CFU/g after 6 and 8 h of exposure to 35 to 37 °C, respectively. All samples became spoiled after 10 h of exposure to 35 to 37 °C, while no enterotoxin was detected in any samples. However, enterotoxins were detected in albacore loin and other samples after 12 and 24 h of incubation at 35 to 37 °C, respectively. Frozen precooked tuna meat should be used for producing canned tuna within 6 to 8 h of thawing to avoid product spoilage and potential enterotoxin production by S. aureus in contaminated precooked tuna meat.     

6种食品防腐剂对金黄色葡萄球菌抑菌效果及肠毒素基因表达的影响

目的：探索6 种食品防腐剂--亚硝酸盐、苯甲酸钠、ε-聚赖氨酸、壳聚糖、茶多酚、Nisin对1 株引起食物中毒的金黄色葡萄球菌SA003的抑菌作用,以及对该菌株所含不同肠毒素基因在mRNA水平表达的影响。方法：按照GB 2760-2014《食品添加剂使用标准》限量标准的质量浓度,分别添加6 种不同的防腐剂在金黄色葡萄球菌SA003初始接菌量约为105 CFU/mL的TSB液体培养基中,37 ℃培养24 h后进行菌落计数;同时收集菌体用于RNA提取,将提取的RNA进行反转录后,采用荧光定量聚合酶链式反应方法检测各肠毒素基因在mRNA水平上的相对表达量。结果：在国标最大限量条件下,不同添加剂对金黄色葡萄球菌SA003的抑菌作用强弱顺序分别为：Nisin＞茶多酚＞壳聚糖＞ε-聚赖氨酸＞苯甲酸钠＞亚硝酸盐。6 种防腐剂均能抑制肠毒素sea、sed、seg、sei、selj、selm、selr和selu基因在转录水平上的表达,但作用效果存在差异。结论：在TSB液体培养基中添加一定剂量的防腐剂不仅能抑制金黄色葡萄球菌的生长,还能够显著降低肠毒素基因的表达量。     

Staphylococcus aureus Growth and Toxin Production in Imitation Cheeses

The ability of eleven imitation (or substitute) cheeses to support the growth and toxin production of Staphylococcus aureus at 26°C was evaluated. All established enterotoxin serotypes were tested by inoculating suspensions of the requisite strains into 100-g samples of cheese (about 30 staphylococci/g). Water activity (a_w) of the cheese samples ranged from 0.942–0.973; pH values ranged from 5.33–6.14. Seven cheeses supported extensive growth of S. aureus; one or more serotypes of enterotoxin were produced in six cheeses. Enterotoxin in the cheese was detected in 4 days at 3 × 10⁶ staphylococci/g. However, the ability of some cheeses to support growth and toxin production of S. aureus could not be correlated with pH, a_w, or product formulation.     

金黄色葡萄球菌肠毒素在食源性微生物中的研究进展

由细菌污染引起的食源性疾病依然是影响人类公共健康和食品安全的最大问题之一。其中,金黄色葡萄球菌是人类的一种重要病原菌,引起许多严重感染。金黄色葡萄球菌属于革兰氏阳性球菌,广泛分布于自然界,是引起化脓性疾病的重要病原菌,也是引起食品污染和细菌性食物中毒的一种重要细菌。食品受金黄色葡萄球菌污染后,不仅会腐败变质,而且部分菌株产生金黄色葡萄球菌肠毒素（Staphylococcal enterotoxins,SEs）而引起食物中毒,由金黄色葡萄球菌肠毒素引起的食物中毒占整个细菌性食物中毒的首位。所以,对SEs的研究以及快速、精确的检测和筛查,成为关键环节。本文拟对金黄色葡萄球菌肠毒素生物学性状、致病．洼、检验方法等方面的研究进展进行综述。当然,对金黄色葡萄球菌及其主要致病因子肠毒素的研究有待进一步深入与发展。     

Model for Combined Effects of Temperature, pH and Water Activity on Thermal Inactivation of Bacillus cereus Spores

Initially, the effect of water activity (a_w) on heat resistance of Bacillus cereus spores (decimal reduction time) was investigated. A linear relationship was found between log D and 1-a_w. The combined effects of temperature (85-105°C), pH (4.5-6.5) and water activity (0.80-1) were then studied. A four parameter model was fitted to the data. This model appeared to be parsimonious with each parameter having a biological significance. Interactions between factors were observed but they accounted for <2.4% of the total variation and they were not taken into account by the model.     

金黄色葡萄球菌新型肠毒素SEK原核表达、纯化及溶液构象分析

金黄色葡萄球菌K型肠毒素（Staphylococcus aureus enterotoxin K,SEK）由金黄色葡萄球菌肠毒素基因簇编码,流行病学显示,编码SEK蛋白的sek基因在食品源金黄色葡萄球菌菌株中具有较高的检出率,说明SEK也可能是一种重要的食品中毒潜在致病因子。本研究将截去N端信号肽的SEK蛋白编码基因与原核表达载体pET-28a（＋）连接,构建重组表达质粒pET-28a（＋）-ΔNspsek;通过对sek基因在不同原核表达宿主菌中的表达及表达条件优化分析,确立SEK蛋白的最优表达条件;利用镍亲合层析纯化含组氨酸（His）标签的SEK融合蛋白,将凝血酶切除His标签后的SEK蛋白进行聚合状态及热稳定分析、荧光发射谱和圆二色谱分析。结果表明,重组蛋白SEK表达成功,热处理导致蛋白部分降解;荧光光谱揭示SEK蛋白在278?nm和295?nm波长处具有相同的最大色氨酸发射峰;344?nm波长处表明SEK蛋白处于紧密折叠的天然状态;圆二色谱分析表明,ΔNspSEK重组蛋白富含β-折叠（23.6%）、β-转角（28%）以及α-螺旋（29.1%）等二级结构。为深入研究SEK蛋白的结构与功能提供基础,也对改进食品加工工艺和提高食品安全具有指导意义。     

Influence of Temperature, pH, Water Activity and Antifungal Agents on Growth of Aspergillus flavus and A. parasiticus

Effect of temperature, pH, water activity, and nine antifungal agents on growth of Aspergillus flavus and A. parasiticus was determined on Sabouraud-Dextrose Agar and on corn. Maximal growth of the two molds occurred at 33°C, the highest temperature used, pH of 5.0 and a_w of 0.99. At 15°C, growth was observed at a_w of 0.95 but not 0.90. Slight growth was observed at an a_w, of 0.85 at 27°C and 33°C. Nine antifungal agents (Botran, Orthocide, Poly-ram 80, Topsin-M, Thiram, Imazalil, sodium propionate, sodium sulfite and DDVP) were tested for inhibition of growth. Activity of the antifungals increased as the a_w was decreased. All antifungals showed inhibitory activity, but Imazalil and DDVP were the most effective agents at the lowest concentrations.