Food safety: new concepts for the new millennium

Current market trends for high quality, fresh, convenient foods, plus improvements in efficiency and reductions in cost mean that the rate of adoption of non-thermal processes is likely to increase. Obstacles to commercialization include the lack of systematic inactivation kinetic data, the interpretation of non-linear death kinetics and the need to establish equivalent control measures for non-thermal treatments in comparison with traditional heat processes. The commercialization of new non-thermal technologies, such as high pressure processing and pulsed electric fields, could be expedited by following guidelines given by the International Commission on Microbiological Specifications for Foods (ICMSF). ICMSF has recently proposed a scheme for the management of microbial hazards for foods, which includes the concept of Food Safety Objectives (FSOs). FSOs are intended to communicate the level of a hazard that is required to meet a given public health goal and to facilitate the acceptance of different, but equivalent processes. ICMSF principles of setting FSOs, the use of performance criteria, process criteria and validation in relationship to HACCP and GHP plans are described. Additionally, the use of FSOs as a framework for developing equivalent control measures are discussed in the context of establishing inactivation regimes based on non-thermal technologies. The implications of non-linear death kinetics to the establishment of process criteria are discussed, especially in relation to the development of safe, equivalent processes for commercial food production.     

An outbreak of food poisoning due to egg yolk reaction-negative Staphylococcus aureus

An outbreak of staphylococcal food poisoning due to an egg yolk (EY) reaction-negative strain occurred in Japan. Twenty-one of 53 dam construction workers who ate boxed lunches prepared at their company cafeteria became ill, and eight required hospital treatment. The outbreak showed a typical incubation time (1.5-4 h with a median time of 2.7 h) and symptoms (vomiting and diarrhea) of staphylococcal food poisoning. Staphylococcus aureus, which produces staphylococcal enterotoxin (SE) A, was isolated from four fecal specimens of eight patients tested. Scrambled egg in the boxed lunches contained 20-40 ng/g of SEA, and 3.0 x 10(9)/g of viable S. aureus cells that produced this toxin. All isolates from patients and the food were EY reaction-negative, coagulase type II, and showed the same restriction fragment length polymorphism (RFLP) pattern. We concluded that the outbreak was caused by scrambled egg contaminated with EY reaction-negative S. aureus. In Japan, outbreaks of staphylococcal food poisoning are mainly caused by EY reaction-positive S. aureus, and EY reaction-negative colonies grown on agar plates containing EY are usually not analyzed further for detection of S. aureus. The present outbreak suggested that EY reaction-negative isolates should be subjected to further analysis to detect the causative agents of staphylococcal food poisoning.     

金黄色葡萄球菌及其引起的食物中毒的研究进展

金黄色葡萄球菌广泛存在于自然环境中,致病力强,是常见食物中毒的病原菌。本文对金黄色葡萄球菌致病因素、污染状况、食物中毒、试验诊断、预防措施等方面进行概述,为今后金黄色葡萄球菌食物中毒的预防提供参考。     

食物中毒事件中致病菌肠毒素检测和分子分型研究

目的 对一起食物中毒事件样本进行检测,查明食物中毒原因,并对分离菌株进行相关性分析。方法 参照国家标准方法GB4789.10-2016,对采集样本进行细菌学检验。应用酶联免疫吸附法(ELISA)检测葡萄球菌肠毒素。通过脉冲场凝胶电泳(PFGE)进行分子分型,分析菌株之间的相关性。结果 食物中毒事件中5名患者洗胃液、食物、分离菌株,均检测出E型葡萄球菌肠毒素。3株分离菌株PFGE分子分型提示来源不同克隆株,除了产生E型葡萄球菌肠毒素外,还有B、C、D型。结论 该起食物中毒由不同PFGE型别的产毒金黄色葡萄球菌污染食物引起,菌株产葡萄球菌肠毒素的型别不完全相同。金黄色葡萄球菌引起的食物中毒应关注多污染源及菌株肠毒素基因表达情况。     

食源性金黄色葡萄球菌肠毒素及其检测方法

金黄色葡萄球菌是一种重要的食源性致病菌,在自然界中广泛分布,其引起的食物中毒是世界性的公共卫生问题。金黄色葡萄球菌肠毒素是引起金黄色葡萄球菌食物中毒的主要致病因子。目前共发现22种金黄色葡萄球菌肠毒素或类肠毒素(SEA-SEE、SEG-SET、SElU、SElU_2和SElV),其中具有催吐活性的被定义为肠毒素,没有催吐活性或者尚待验证的被定义为类肠毒素(SEl)。传统肠毒素SEA~SEE被报道是引起金黄色葡萄球菌食物中毒的主要肠毒素类型,但是大多数新型肠毒素或类肠毒素与食物中毒的关系还没有被真正认识。本文对近几年关于金黄色葡萄球菌肠毒素与食物中毒的关系、肠毒素的表达调控以及肠毒素检测方法的研究进展进行了总结,以期更好地了解金黄色葡萄球菌新型肠毒素致病性,为今后金黄色葡萄球菌食物中毒的预防和控制提供依据。     

Distribution of newly described enterotoxin-like genes in Staphylococcus aureus from food

Extensive analysis of the Staphylococcus aureus genome has allowed the identification of new genes encoding enterotoxin-like superantigens (SEls). Some of these are thought to be involved in staphylococcal food poisoning, while others do not elicit any emetic effect. The potential impact of these members of the enterotoxin-like family on the human organism seems to rely mainly on their superantigenic activity. In this paper the distribution of the genes coding for enterotoxin-like superantigens in S. aureus isolated from food was studied. Fifty isolates of S. aureus were examined and 27 were shown to be enterotoxigenic. Only 9 of the 27 strains carried genes encoding enterotoxins SEA-SEE. In 18 SEA-SEE-negative strains the presence of newly described enterotoxin genes was detected. All SEA-SEE-positive strains simultaneously carried genes of new SEls. We show that the gene encoding SElH (staphylococcal enterotoxin-like enterotoxin H) was the most frequently detected (n=14), while genes encoding SElI together with SElG accompanied by the other genes of the egc locus were detected in three strains. We also detected the presence of three less investigated genes: sep, sel, and sek. These genes were present in eight, two, and one isolate, respectively. In one strain, sep was accompanied by genes of other SEls, while in the remaining seven it was the only enterotoxin-like gene detected. The high prevalence of newly discovered enterotoxin genes, including the genes encoding emetic toxins, was demonstrated in food-derived strains. This supports the need for additional work on its role in food poisoning and, alternatively, to monitor its presence in S. aureus isolated from food. Our results suggest that yet unknown genetic elements encoding enterotoxin genes may exist.     

我国食品卫生微生物标准体系中存在的问题及建议

主要对我国的食品卫生微生物标准体系存在问题进行分析,借鉴食品法典委员会(CAC)、国际食品微生物规格委员会(ICMSF)及发达国家的食品卫生微生物标准体系,结合我国的国情,提出修改我国食品卫生微生物标准体系的构想和建议。     

2015~2016年安康市食物中毒检测分析

目的 回顾分析2015~2016安康市食物中毒事件, 总结安康市事物食物中毒特点。方法 收集安康市的中毒事件的纸质材料进行分析。参考GB 4789-2016《食品卫生微生物学检验》、WS271-2007《感染性腹泻诊断标准》和WS289-2008《霍乱诊断标准》等试验方法对食物中毒样本进行检测。结果 2015~2016年安康市发生食物中毒事件14起, 1起为有机磷中毒, 其余13起均为食源性致病菌中毒。检出致病菌4种16株, 分别是: 沙门氏菌4株; 金黄色葡萄球菌5株、蜡样芽胞杆菌3株; 致泻性大肠埃希氏菌EPEC-A、EHEC、EAEC、EIEC各1株。 结论 安康市食物中毒多以细菌性为主, 污染的致病菌种类多且致病性强。     

国内外食品卫生微生物标准体系探究

主要对国内外食品卫生微生物标准体系中的食品种类、检验项目、限量标准、应用要求、取样计划等内容进行比较分析,指出我国食品卫生微生物标准体系与食品法典委员会(CAC)、国际食品微生物规格委员会(ICMSF)及发达国家相比存在的差距和不足,提出了完善我国食品卫生微生物标准体系的建议。     

Interactions between Staphylococcus aureus and lactic acid bacteria: an old story with new perspectives

Staphylococcus aureus is a gram positive opportunistic pathogen and a major concern for both animal and human health worldwide. In some contexts where Lactic Acid Bacteria (LAB) are the normal dominant microbiota, such as in fermented food or in the vaginal ecosystem, S. aureus sometimes colonises, persists, expresses virulence factors and produces food poisoning or urogenital infections, respectively. Studies on the interactions between LAB and S. aureus began a few decades ago and were pursued to shed light on the inhibitory capabilities that LAB might have on S. aureus growth and/or enterotoxin production in fermented foodstuffs. These early studies had the aim of developing methods to prevent staphylococcal food poisoning, thus improving food safety. More recently, the concept of vaginal probiotic LAB has emerged as a promising way to prevent urogenital infections, S. aureus being one of the potential pathogens targeted. This review provides an up-to-date look at the current hypotheses of the mechanisms involved in the inhibition of S. aureus by LAB in both the vaginal ecosystem and in fermented food ecosystems. We also emphasise that post-genomic approaches can now be envisioned in order to study these diverse and complex interactions at the molecular level. Further works in this field will open up new avenues for methods of biocontrol by LAB and/or for biotechnological uses of LAB-compounds to fight against the long-standing, yet incumbent menace of staphylococcal infection.     

Characteristics of enterotoxin H-producing Staphylococcus aureus isolated from clinical cases and properties of the enterotoxin productivity

Staphylococcal enterotoxin H (SEH) is predicted to be involved in staphylococcal food poisoning. To characterize SEH-producing Staphylococcus aureus isolates from staphylococcal food poisoning cases in Japan, we investigated the relationship between SEH production and coagulase serotype, which is an epidemiological marker, and compared the properties of SEH production with those of staphylococcal enterotoxins A (SEA) and B (SEB). SEH production was determined by a newly developed sandwich enzyme-linked immunosorbent assay. Eighty-six (59.7%) of 144 isolates from staphylococcal food poisoning cases produced SEH. Seventy-one of the SEH-producing isolates simultaneously produced SEA, SEB, or both. All SEH-producing isolates belonged to coagulase type VII, which was the predominant type, representing 99 (68.8%) of 144 isolates. The amount of SEH produced in brain heart infusion was almost the same as the amount of SEA and approximately 10-fold lower than that of SEB. SEH and SEA were produced mainly during the late exponential phase of growth, whereas SEB was produced mostly during the stationary phase. The production levels of SEH and SEA were gradually affected by decreases in water activity, but the production of SEB was greatly reduced under conditions of low water activity. These findings indicate that SEH-producing S. aureus isolates are of high prevalence in staphylococcal food poisoning cases. Given the unique epidemiological characteristic of these isolates, SEH and SEA probably are responsible for food poisoning.     

Use of novel PCR primers specific to the genes of staphylococcal enterotoxin G, H, I for the survey of Staphylococcus aureus strains isolated from food-poisoning cases and food samples in Taiwan

Data regarding the incidence of the newly found enterotoxigenic Staphylococcus aureus strains in food poisoning cases and in food samples were to date not available in Taiwan. In this study, PCR primers specific for the detection of SEG, H and I genes, i.e., seg, seh and sei, were used for the assay of 55 human isolates of S. aureus negative to the classical enterotoxins (SEA-->SEE) detection. These isolates were from the fecal specimens of the patients suffering from food poisoning outbreaks. Only eight strains were found to have the seg, seh and sei. The presence of other bacterial pathogens, such as Vibrio parahaemolyticus, Bacillus cereus and Salmonella spp. and perhaps, strains producing other new staphylococcal enterotoxins, in the fecal specimens of these patients, may account for these food poisoning cases. For 139 strains from food samples, such as frozen Chinese foods, Chinese sausages and lunch meals, sea strains accounted for the major portion and it seemed to be the most common SE type to coexist with seg, seh and sei. Only two strains had sec and none of them had seg, seh or sei. For strains without the classical SE genes, only 13 strains had seg, seh and/or sei. The above results imply that seg, seh and sei S. aureus strains play only a minor role in food-borne outbreaks in Taiwan.     

Applications of chia (Salvia hispanica L.) in food products

BackgroundThe outstanding nutritional and technological properties lead to innovative applications of chia (Salvia hispanica L.) in food products.Scope and approachThe aim of this contribution is to give an overview of the various food application approaches for chia. The nutritional and technological properties of chia as well as its technological and innovative utilization are presented. Examples for the various applications in food products are given in five main topics: baked goods, dairy products, meat and fish products, gluten-free products and other products such as functional food, hydrocolloid and thickener.Key findings and conclusionsBesides the nutritional benefits of chia incorporation in food products the technological effects are emphasized very often. Summarized in five main topics, most authors conclude that chia is a valuable food ingredient for functional food development. The application of chia in baked goods for example is beneficial not only to improve the nutritional value but acting as hydrocolloid or substitute egg, fat or gluten. The increase of oil stability and applications as food thickener in novel food applications are of high importance too. As different chia fractions give various options for applications the utilization of chia for further food products will increase significantly in the future.     

一起食物中毒事件中产独特肠毒素表型的金黄色葡萄球菌病原学分析

目的对一起疑似为金黄色葡萄球菌所导致的食物中毒事件进行葡萄球菌肠毒素检测,结合金黄色葡萄球菌病原学分析,为明确食物中毒诊断提供依据。方法根据流行病学调查,采用ELISA方法对可疑食物进行葡萄球菌肠毒素检测,同时对可疑食物和患者呕吐物进行金黄色葡萄球菌分离,运用Vitek2 Compact全自动细菌鉴定仪和血浆凝固酶试验鉴定为金黄色葡萄球菌,采用脉冲场凝胶电泳(PFGE)对病原菌进行同源性分析,以ELISA方法对检出的金黄色葡萄球菌菌株进行肠毒素检测,用PCR方法对肠毒素基因进行分型。结果食物和患者样品中分别分离出2和11株金黄色葡萄球菌,PCR方法及ELISA方法对肠毒素分型结果显示,其中12株同时存在SEA、SEB、SED、SEE 4种肠毒素及相关基因,PFGE聚类分析显示,其中12株产肠毒素金黄色葡萄球菌具有高度同源性。结论本起食物中毒事件为具有独特肠毒素表型的金黄色葡萄球菌导致,在金黄色葡萄球菌中毒实验室调查过程中,肠毒素检测结合病原菌溯源分析可以为相关公共卫生事件提供科学依据。     

食品中金黄色葡萄球菌概况及新型检测技术研究进展

金黄色葡萄球菌是食品中常见的食源性致病菌,是引起细菌性食物中毒最普遍的原因之一,感染后可引起人食物中毒、急性肠胃炎和动物腹泻等疾病,严重危害人类安全及健康。因此研究食品中金黄色葡萄球菌的特性及快速准确的检测方法、对及时有效控制病原菌传播、预防食物中毒,减少食源性疾病事件的发生具有重要的现实意义。     

Foodborne outbreak caused by Staphylococcus aureus: phenotypic and genotypic characterization of strains of food and human sources

An outbreak of staphylococcal food poisoning involving approximately 180 people occurred in Brodowski, S?o Paulo State, Brazil, in April 1998. Strains of Staphylococcus aureus isolated from foods and food handlers, implicated as the etiologic agent, were characterized with phenotypic (phage typing, antibiotic susceptibility test, and enterotoxin production), and genotypic (random amplified polymorphic DNA) characterization. Strains isolated from vegetable salad with mayonnaise sauce, broiled chicken, pasta in tomato sauce, and from the oropharyngeal secretions of five food handlers--A, B, C, H, and I--showed the same phage profile and antibiotic resistance. Random amplified polymorphic DNA generated 17 combined profiles with primers OPE-20 and OPA-7. The similarity of strains was analyzed by generating a dendrogram that classified the 59 strains of S. aureus into four major clusters (I, II, III, and IV). Strains from four food handlers (A, B, H, and I) and from vegetable salad with mayonnaise, broiled chicken, and pasta in tomato sauce showing the same phage type profile and resistance to antibiotics belonged to the same cluster and produced staphylococcal enterotoxin A. Therefore, these foods and food handlers were incriminated in the outbreak.     

Ribotyping and a study of transmission of Staphylococcus aureus collected from food preparation facilities

Food poisoning from Staphylococcus aureus is sometimes caused by improper handling of food items in food preparation facilities. Prevention of contamination by employees is particularly important in facilities where a significant amount of food preparation is performed by hand. Some experiments have been performed to describe bacterial cross-contamination in the food preparation process, but there have been few studies of cross-contamination in actual food preparation facilities. Aiming to shed light on the transmission of S. aureus in food preparation facilities, this study collected samples of 66 strains of this bacterium from the fingers of food preparation staff, foodstuffs, prepared foods, cooking utensils, and cooking equipment and typed them with the ribotyping method. S. aureus from the same ribogroup was detected on the hands of a study participant, a faucet, knife, frying pan, and a salad, indicating that bacteria found on the hands of the study participant was transmitted to cooking utensils and prepared foods. Transmission (from a faucet to a frying pan handle) of bacteria by another person, a third party, was also detected.     

The detection of enterotoxins and toxic shock syndrome toxin genes in Staphylococcus aureus by polymerase chain reaction

A simple polymerase chain reaction (PCR)-based procedure was developed for the detection of fragments of staphylococcal enterotoxins (SEs) SEA, SEB, SEC, SED, SEE, SEG, SEH, and SEI together with the toxic shock syndrome toxin (TSST-1) genes of Staphylococcus aureus. One hundred and twenty-nine cultures of S. aureus were selected, 39 of which were recovered from 38 suspected staphylococcal food-poisoning incidents. The method was reproducible, and 32 different toxin genotypes were recognized. The presence of SE genes was associated with S. aureus strains reacting with phages in group III, and the TSST-1 gene with phages in group I. There was a 96% agreement between the PCR results for detection of SEA-D and TSST-1 as compared with a commercial reverse passive latex agglutination assay for the detection of SEs from cultures grown in vitro. Enterotoxin gene fragments were detected in S. aureus cultures recovered from 32 of the 38 suspected staphylococcal food poisoning incidents, and of these, 17 were associated with SEE, SEG, SEH, and SEI in the absence of SEA-D. Simple PCR procedures were also developed for the detection of SE directly in spiked food samples, and this was most successfully achieved in mushroom soup and ham. Detection was less successful in three types of cheese and in cream. SEA or SEB were detected by enzyme-linked immunosorbent assay in three food samples (two of which were associated with food poisoning incidents) naturally heavily contaminated with S. aureus: the appropriate SEA or SEB gene fragments were detected directly in these three foods by PCR.     

生菜生产到消费全程金黄色葡萄球菌的定量风险评估

目的：评估即食生菜金黄色葡萄球菌（Staphylococcus aureus）引起食物中毒的风险,为风险管理措施的选择提供理论支持。方法：排查生菜在大棚和露地两种种植方式下各环境样本的污染率及污染水平,结合居民消费习惯和生菜中金黄色葡萄球菌的生长模型,描述生菜从农田到餐桌过程中金黄色葡萄球菌的变化,从危害识别、危害特征描述、暴露评估和风险特征描述4 方面评估我国即食生菜导致金黄色葡萄球菌中毒的风险。结果：大棚生菜的土壤中有金黄色葡萄球菌检出,检出率为0.04%,污染水平为－8.72～3.01（lg（CFU/g））,经过灌溉、采收、运输、销售、购买、贮藏及清洗等过程,生菜中金黄色葡萄球菌在食用前的污染平均值为－1.95（lg（CFU/g））,90%置信区间为－8.98～4.40（lg（CFU/g））。评估结果表明,我国因即食生菜导致金黄色葡萄球菌中毒的概率为2.72×10－4,每年因此引发的食物中毒病例约为77万 人。敏感性分析结果显示,土壤中金黄色葡萄球菌的初始污染水平对生菜中金黄色葡萄球菌的污染水平影响最大,其次为家庭贮藏温度及食用前清洗过程;控制生菜种植卫生环境、合理使用冰箱及清洗生菜可有效降低即食生菜引起金黄色葡萄球菌中毒的风险。