Production of diarrheal toxin by selected strains of Bacillus cereus

The production of diarrheal toxin by six selected strains of Bacillus cereus was monitored during growth at 32 °C, a temperature described as near-optimal for growth and toxin production. Toxic activity was measured in culture filtrates and cellular extracts sampled at three different times during growth. Two alternative methods, a cytotoxicity test on Chinese hamster ovary (CHO) cells and a commercial immunological test (BCET-RPLA, Oxoid) were used. Toxin titres were in agreement with epidemiological characteristics and toxicity demonstrated by using other systems in other examinations. A comparison of intra- and extracellular toxicities measured at the exponential and stationary growth phases showed that the toxin was essentially secreted during the exponential phase. For several strains, secretion peaked during the period from the middle exponential phase to the beginning of the stationary phase. There was no important overall increase of the toxicity during full and late stationary phase. The level was stable or even lower, thus indicating that diarrheal toxin production during stationary phase was small, if any, and that the toxin was unstable under these conditions. Statistical analysis of toxicities showed that the cytotoxicity test was correlated with the immunological test (significant at a 1% level). For routine determinations, a toxicologic laboratory may use any of the two methods, depending oft its facilities, the immunological test being relatively expensive.     

生食蔬菜模拟污染米饭样本中蜡样芽胞杆菌和呕吐毒素基因分布状况研究

目的 分析被生食蔬菜模拟污染的米饭样本中蜡样芽胞杆菌（Bc）和呕吐毒素基因（ces）的分布状况,为Bc食物中毒科学防控提供基础数据。方法 采集生食蔬菜样本50件,每件样本用0.85%生理盐水盥洗后污染“新煮熟米饭”,置于30 ℃、70% RH培养箱中放置24 h。对生食蔬菜和“污染米饭”进行Bc的定量计数、荧光PCR检测和数字PCR检测。对基于不同采集地点、不同蔬菜类型分组的生食蔬菜样本和及被其污染的“污染米饭”的各项检出率指标进行统计学分析。结果 生食蔬菜样本中Bc检出率为80.00%（40/50）,ces基因和Bac16s RNA基因检出率分别为0（0/50）和10.00%（5/50）;“污染米饭”样本Bc检出率为94.00%（47/50）,ces基因和Bac16s RNA基因检出率分别为14.00%（7/50）和90.00%（45/50）。采集自农贸市场和农户土地的2组生食蔬菜中Bc检出率差异有统计学意义（χ²=11.063,P=0.000 88校正）,被上述2组生食蔬菜类污染的“污染米饭”Bac16s RNA基因检出率和ces基因检出率差异均有统计学意义（χ²=3.926,P=0.047 5校正;χ²=5.444,P=0.019 6校正）。7件“污染米饭”基于荧光PCR检测ces基因阳性,Ct值介于24.12~37.73,数字PCR结果介于6.8 copes/μL~6.2×10⁶ copes/μL。结论 被生食蔬菜模拟污染的米饭样本可具有导致Bc食物中毒风险的病原学特征。     

2014—2019年岳阳市食源性蜡样芽胞杆菌溶血素hblA与磷脂酶C plc基因特征分析

目的 了解湖南省岳阳市2014－2019年食源性蜡样芽胞杆菌菌株病原学特征,并为蜡样芽胞杆菌引起的食物中毒事件的科学防控提供依据。方法 对2014—2019年分离自湖南省岳阳市城区餐饮门店的26株蜡样芽胞杆菌菌株的致病毒力因子溶血素BL的hblA基因和磷脂酶C的plc基因进行序列扩增和测序,并使用Seqman和MEGA X软件对蜡样芽胞杆菌的hblA和plc基因进行遗传进化分析。结果 从岳阳市分离到的蜡样芽胞杆菌hblA、plc毒力基因的同源性与GenBank中的蜡样芽胞杆菌群相比均大于93.0%。结论 岳阳市分离的蜡样芽胞杆菌的hblA和plc基因与GenBank中的蜡样芽胞杆菌群的同源性高,具有一定的亲缘关系。本研究为进一步了解和科学防控蜡样芽胞杆菌引起的食物中毒事件奠定了研究基础。     

蜡样芽孢杆菌LJ01中亚硝酸盐还原酶的基因克隆和生物信息学分析

为研究蜡样芽孢杆菌LJ01(Bacillus cereus LJ01)中亚硝酸盐还原酶(NiR)的酶学性质,获得高表达量的基因工程菌,本文对B.cereus LJ01的NiR序列进行了生物信息学分析,并将NiR基因克隆到表达载体pET-28a(+)和pET-32a(+)中,构建了基因工程菌pET-28a(+)-nir-BL21和pET-32a(+)-nir-BL21,随后探究了不同诱导条件对重组NiR表达量的影响。结果表明NiR编码蛋白的理论分子量约为60 kDa,理论pI为5.47,二级结构主要为α-螺旋和无规则卷曲,是不具有跨膜结构的亲水性蛋白。随着诱导温度的升高,重组NiR的表达量逐渐减少,诱导温度为16℃时重组NiR表达量最高。在同一诱导温度下,NiR在pET-28a(+)-nir-BL21中的表达量明显高于pET-32a(+)-nir-BL21,因此选用pET-28a(+)-nir-BL21作为基因工程菌。从B.cereus LJ01中克隆了NiR基因,构建了基因工程菌pET-28a(+)-nir-BL21,为该重组NiR的理化性质研究和食源芽孢杆菌中NiR的异源表达奠定了基础。     

阿魏酸对大肠杆菌和蜡状芽孢杆菌产组胺的影响

以产组胺的大肠杆菌(E. coli)和蜡状芽孢杆菌(B. cereus)两株菌为研究对象,通过双倍琼脂稀释法,得出阿魏酸对其最低杀菌浓度(MBC)和最低抑菌浓度(MIC);并利用高效液相色谱法(HPLC)检测不同浓度的阿魏酸对两株菌株产组胺的影响,从而明确阿魏酸对其产组胺的抑制作用。结果表明:阿魏酸对供试菌株都具有较强的抑菌活性,且抑菌效果随着阿魏酸浓度的增大而增强;阿魏酸对E. coli的MIC为0.031%,对B. cereus的MIC为0.063%;当阿魏酸的添加量为MIC时,E. coli和B. cereus的生长均受到抑制,导致组胺含量显著降低(p<0.05),E. coli和B. cereus所产组胺量较空白对照组分别降低了18.45%和25.78%。所以,阿魏酸是一种能够显著抑制组胺的优良添加物,在食品工业中具有很好的应用前景。     

Properties of Bacillus cereus and other bacilli contaminating biomaterial-based industrial processes

This paper is an overview on bacilli in industrial processes, with focus on food grade paper and paperboard production. Paperboards mainly contain sporeforming bacteria belonging to the genera Bacillus, Paenibacillus and Brevibacillus, usually found in quantities from <50 to 250 cfu g⁻¹ homogenized paperboard. Of those frequently found, Bacillus cereus group, B. licheniformis, B. subtilis and Brevibacillus brevis are important for food hygiene because of their hydrolytic activities on food components and the ability of some strains to produce food poisoning toxins or to grow at refrigerated temperatures. We found that the phenotypic properties (lecithinase activity, nitrate reduction) used in standard methods (e.g., ISO, FDA, IDF) to recognize B. cereus, were unreliable for industrial isolates. Whole cell fatty acid composition of a group of the industrial isolates deviated so much from those in a widely used commercial database that the strains were not or only poorly recognized as B. cereus. Industrial isolates, including toxigenic ones, often missed one or more of these characters, even in cases where 100% 16S rDNA identity was found with B. cereus or with B. thuringiensis. 11-Methyldodecanoic acid and trans-9-hexadecenoic acid were found without exception in over 200 industrial B. cereus group isolates and in over 30 culture collection strains. The detection of these fatty acids is a secure method for the identification of B. cereus. Negative reaction for starch hydrolysis and for BCET-RPLA test and a specific ribotype were found in all B. cereus strains producing the emetic toxin.     

即食米面制品中蜡样芽胞杆菌分离鉴定及毒力基因研究

为探究即食米面制品中蜡样芽胞杆菌的污染状况及食源性蜡样芽胞杆菌中毒力基因的分布规律,以餐饮服务场所采集245份即食米面制品为蜡样芽胞杆菌的污染调查材料,通过国标法及持家基因分离鉴定得到49株蜡样芽胞杆菌阳性株,并对其进行13种毒力基因的PCR检测。结果表明:蜡样芽胞杆菌的平均检出率为10.61%(26/245),阳性检出样品主要为盒饭及米饭,其蜡样芽胞杆菌平均检出水平为3032 CFU/g,其中以盒饭的检出程度最高。49株分离菌株均检出两种或两种以上的毒力基因,非溶血性肠毒素基因nhe和entFM检出率最高,分别达到100%及91.84%,是分离菌株的主要毒力基因;溶血素基因hblA、hblC、hblD检出率分别为20.41%、38.78%和40.82%。同时携带nhe三个基因又携带hblA、hblC和hblD基因的强毒株有7株,占14.29%,且这7株菌均含有entFM基因。呕吐型毒力基因ces、cer、EM1仅2株检出,检出率为4.08%。本研究对即食米面制品中蜡样芽胞杆菌的监控、预警及爆发引起食物中毒后追踪其感染源和传播途径及构建基因指纹图谱库和分子溯源平台具有指导意义。     

活性氧与蜡样芽孢杆菌菌膜形成的相关性

为阐明活性氧（ROS）与蜡样芽孢杆菌菌膜形成的相关性,以市售原料奶中分离的蜡样芽孢杆菌分离株为目标菌,以细菌中NADPH氧化酶介导产生的ROS为主要靶点,用外源ROS补充剂过氧化氢(H₂O₂)、ROS清除试剂N-乙酰半胱氨酸（NAC）、NADPH氧化酶抑制剂二苯基氯化碘盐（DPI）处理蜡样芽孢杆菌,测定分析ROS变化与菌膜形成之间的关系。结果表明,0.01 μmol/L H₂O₂、1 μmol/L H₂O₂、100 μmol/L H₂O₂处理组平均菌膜形成量均显著高于对照组平均菌膜形成量（P<0.05）,0.1 μmol/L H₂O₂、10 μmol/L H₂O₂处理组平均菌膜形成量与对照组平均菌膜形成量相比无显著性差异（P>0.05）,随着H₂O₂浓度的升高,ROS逐渐下降。NAC各处理组平均菌膜形成量均显著高于对照组平均菌膜形成量（P<0.05）,随着NAC浓度的升高,ROS逐渐下降。DPI浓度≤1 μmol/L时,随着DPI浓度的升高,平均菌膜形成量均显著高于对照组平均菌膜形成量（P<0.05）,ROS逐渐下降;DPI浓度>5 μmol/L时,随着DPI浓度的升高,平均菌膜形成量显著低于对照组平均菌膜形成量（P<0.05）,ROS含量升高。表明在不影响蜡样芽孢杆菌生长和细胞活性的前提下,一定浓度的H₂O₂、NAC和DPI处理菌株能够诱导ROS减少,增强蜡样芽孢杆菌菌膜的形成。激光共聚焦显微镜结果显示,与对照组相比处理组具有更强的染色效果,呈现红色的网络结构,而未经处理组呈现松散的结构和更少的菌膜生物量,这表明ROS对蜡样芽孢杆菌菌膜的形成存在抑制作用。     

蜡样芽孢杆菌高密度发酵条件与过程的优化

蜡样芽孢杆菌的生物量对其生物功能具有重要影响。为了提高蜡样芽孢杆菌的产量与芽孢率,通过单因素实验筛选出最适宜其生长的碳源与氮源,分别为质量比1∶1复配的葡萄糖与水溶性淀粉和质量比1∶1复配的大豆蛋白胨与酵母提取物。在此基础上,于7 L发酵罐中探究流加方式、p H、搅拌转速和通气量等因素对其生物量及芽孢率的影响。最优发酵条件为：在0～9 h之间,转速250 r/min,通气量3 L/min,pH恒定6.5;9～18 h时,转速升高至350 r/min,通气量升高至4.5 L/min,并以0.45 mL/min流量补加培养基浓缩液;18 h时,转速降低至150 r/min,通气量降低至2.25 L/min,pH调高至7.5,停止补料至发酵结束。基于以上发酵策略,在18 h时,蜡样芽孢杆菌活菌数达2.2×10¹⁰CFU/mL,是未优化前的4.88倍;发酵结束时,芽孢率超过90%。本研究结果为蜡样芽孢杆菌的工业化应用提供了一定基础。     

婴幼儿配方乳粉加工环境中蜡样芽孢杆菌多位点序列分型

以分离自婴幼儿配方乳粉加工环境中蜡样芽孢杆菌为研究对象,利用多位点序列分型(MLST)技术对蜡样芽孢杆菌的多样性和系统进化关系进行探究。结果表明,84株蜡样芽孢杆菌共分成24个ST型,分别是ST-24、ST-26、ST-32、ST-62、ST-144、ST-374、ST-999、ST-1119、ST-1243、ST-1284、ST-1333、ST-1334、ST-1335、ST-1336、ST-1337、ST-1338、ST-1339、ST-1340、ST-1341、ST-1342、ST-1343、ST-1344、ST-1345及ST-1347,其中ST-999(22.62%)、ST-1343(15.48%)、ST-1335(7.14%)与ST-1345(7.14%)是该婴幼儿配方乳粉加工环境中的优势ST型。同时发现了3个新的等位基因pur-242,pyc-198,ilv-276与14个新的ST型ST-1333、ST-1334、ST-1335、ST-1336、ST-1337、ST-1338、ST-1339、ST-1340、ST-1341、ST-1342、ST-1343、ST-1344、ST-1345和ST-1347。系统发育分析表明24个ST型与B.cereus、B.anthracis和B.thuringiensis这三个种显示了更近的系统发育关系,与蜡样芽孢杆菌群体中的另外8个种的亲缘关系较远。     

辣木籽提取物提取工艺优化及其对乳中蜡样芽孢杆菌的抑制作用

为了探究辣木籽提取物对乳中蜡样芽孢杆菌的抑菌活性及抑菌稳定性。以抑菌圈为参照,采用单因素和响应面优化辣木籽抑菌成分的提取工艺条件,以温度、pH、紫外线照射衡量抑菌稳定性,并探究辣木籽提取物在液态乳贮藏中的应用。结果表明,辣木籽抑菌成分的最佳提取工艺条件为:以水为溶剂,pH3.1,料液比1:70 g/mL,浸提温度50 ℃,浸提时间2.5 h,在此工艺条件下,辣木籽提取物的抑菌圈大小为(27.23±0.58) mm,最小抑菌浓度（MIC）和最小杀菌浓度（MBC）分别为1.25、5 mg/mL,不同pH及紫外线照射时间处理对辣木籽提取物抑菌活性影响不显著（P>0.05）,以100 ℃加热30 min后其抑菌活性仍可保留空白组的50.97%。在经巴氏杀菌的全脂乳和脱脂乳中添加MIC辣木籽提取物对乳风味、色度和黏度均无显著影响（P>0.05）,且4 ℃条件下可有效抑制蜡样芽孢杆菌的生长,相较普通巴氏杀菌乳可将其保质期由7 d至少延长到14 d。因此,辣木籽提取物对乳中蜡样芽孢杆菌有高效、稳定的抑菌效果,具有应用于液态乳贮藏保鲜的潜力。     

生鲜食品中蜡样芽孢杆菌毒力基因及耐药性研究

目的:研究生鲜食品中蜡样芽孢杆菌的毒力基因及耐药性。方法:在成都市周边农贸市场和路边小摊采集各种生鲜食品共100份,采用MYP选择性培养基初步分离蜡样芽孢杆菌菌株,采用16S rRNA序列比对分析鉴定得到蜡样芽孢杆菌分离菌株,采用PCR检测分离鉴定菌株中蜡样芽孢杆菌13个毒力基因的携带情况,进一步采用纸片扩散法检测菌株对18种抗生素的耐药性。结果:从100份生鲜食品样本中,检出蜡样芽孢杆菌24株,检出率为24%,其中,路边小摊的检出率(70%)高于农贸市场(18.9%)。呕吐型基因ces和cer的检出率较低,仅在1株中发现;腹泻型基因nheC有24株检出;cytK有13株检出;bceT有12株检出;hblA有11株检出;nheB有10株检出;hblC、hblD、nheA各有9株检出;hly-Ⅱ有7株检出;hblB、entFM检出率为0;蜡样芽孢杆菌的4个看家基因rpoB、gyrB、groEL、vrrA在24株分离菌株中检出率为100%。同时发现,生鲜食品来源的蜡样芽孢杆菌分离菌株对杆菌肽B、磺胺甲亚唑、苯唑西林、青霉素表现出较高耐药性,耐药率大于96%;而对阿米卡星、氯霉素、庆大霉素、亚胺培南耐药率小于7%。结论:本研究通过分离鉴定生鲜食品源蜡样芽孢杆菌,研究其毒力基因携带及耐药性,为进一步评价生鲜食品中蜡样芽孢杆菌的安全风险提供参考数据。     

一起疑似由蜡样芽胞杆菌引起的食源性疾病事件的分子溯源研究

目的 对一起食源性疾病事件中食品分离蜡样芽胞杆菌（B. cereus）进行溯源分析,为确定污染源、切断传播途径提供技术支撑。方法 本研究建立脉冲场凝胶电泳（PFGE）方法,对12株B. cereus进行分子分型,同时对10株B. cereus进行全基因组测序（WGS）,利用BioNumerics软件对测序数据进行拼接组装、多位点序列分型（MLST）、毒力基因、核苷酸多态性（SNP）分析。结果 本起食源性疾病事件中分离自不同食品的12株B. cereus的PFGE分型显示为8种带型,其中3株ST1435型 B. cereus带型相同,且SNP分析显示这3株B. cereus只有3个碱基差异;2株ST24型B. cereus 的PFGE带型完全相同,且SNP分析显示只有1个碱基差异,提示3株ST1435和2株ST24菌株分别为克隆株。2株ST24型B. cereus携带溶血性肠毒素hlbA、hlbC和hlbD,另有4株B. cereus携带腹泻毒力基因（hlbA、hlbC和hlbD）。结论 B. cereus引起的食源性疾病事件比较复杂,污染源也比较复杂,因此加强原辅料监测、从业人员的卫生监测、环境和设备及环节的清洗消毒,对预防控制由其引起的食源性疾病事件非常重要。     

Investigation of Bacillus cereus contamination sites in a dairy plant with RAPD-PCR

The usefulness of RAPD-PCR as a typing method for investigation of Bacillus cereus isolates from a dairy plant was evaluated. A 1-d in-depth sampling study along the entire production line was performed and long-term sampling of products was conducted over two years. In addition, farm bulk tank isolates were collected. Isolates from an individual sample during storage at 7°C were often identical. Large clusters of identical isolates were found in the silo tank in the 1-d study and in samples from the long-term study. In combination with analytical data, these results indicated a prolonged contamination problem caused by a mesophilic strain early in the production chain. Detailed and useful information on occurrence of psychrotrophic B. cereus was obtained along the production line and among products. RAPD-PCR appeared to be a useful typing method for B. cereus isolates. In combination with carefully planned sampling and microbiological analysis, RAPD could be helpful in solving dairy contamination problems in pasteurized milk.     

Real-time-PCR-System zum Nachweis von Bacillus cereus (emetischer Typ) in Lebensmitteln

Bacillus cereus is a Gram-positive, facultative anaerobic, spore-performing bacterium. Some B. cereus strains have the ability to produce two different types of toxins: (a) diarrhoeic toxin: the disease is similar to a C. perfringens toxin-infection; it is caused by a heat-labile protein, (b) emetic toxin: the disease is similar to Staphylococcus-aureus intoxikation; it is caused by a heat-stable protein. Statements about the frequency of B. cereusfood-poisonings are difficult because a reporting system for this disease is missing in Germany and there exists no valid methodology to diagnose this disease with conventional microbiological or biochemical methods. Nevertheless B. cereus, beside S. aureus, seems to be the most important bacterium to cause food-associated intoxications.     

四种不同接触表面蜡样芽孢杆菌菌膜形成的影响

为了解食源性致病菌蜡样芽孢杆菌在食品加工环境中菌膜形成能力,以玻璃、不锈钢、聚氯乙烯、聚丙烯为接触面,采用超声波平板菌落计数法测定不同环境因素(温度、pH、氯化钠、葡萄糖、苯甲酸钠及山梨酸钾)、不同材料表面蜡样芽孢杆菌(B.cereus)菌膜形成的变化趋势。结果表明:四种材质表面形成B.cereus菌膜能力的大小顺序为:玻璃 > 不锈钢 > 聚氯乙烯 > 聚丙烯。其中,30 ℃,pH7.0时菌膜形成量最大,添加低浓度葡萄糖(4.0%)或氯化钠(0.5%)对B.cereus菌膜形成有显著促进作用(p<0.05),添加0.15%苯甲酸钠、山梨酸钾的菌膜形成量显著高于添加0.10%的菌膜形成量(p<0.05)。本研究为蜡样芽孢杆菌风险评估提供基础数据,为食品工业蜡样芽孢杆菌菌膜的预防和控制奠定基础,为改进蜡样芽孢杆菌的清洗控制措施提供参考。     

Production of high degree polymerized chitooligosaccharides in a membrane reactor using purified chitosanase from Bacillus cereus

Crude chitosanase from Bacillus cereus NTU-FC-4 was separated by a cation exchanger to three fractions named CBCI, CBCII, and CBCIII. The CBCI hydrolyzed chitosan to yield dimers. The primary hydrolytic products of CBCII were low degree polymerized (DP) chitooligosaccharides. The CBCIII had the fastest reaction rate and yielded high DP chitooligosaccharides (heptamer and higher DP oligomers). When CBCIII was used in the ultrafiltration membrane reactor with enzyme/substrate ratio 0.06 unit/mg and 100 min of residence time (RT), the concentration of high DP oligomers was 9.78 mg/mL which occupied ca. 48% of total oligomers in the final product as compared to ca. 29% resulted from the crude enzyme. Decrease of RT to 50 min and 33 min, the high DP oligomers in the products were ca. 61% and 69%, respectively. This system could be operated for at least 24 h and kept a constant permeate flux and product output rate.     

1株食源性蜡样芽胞杆菌分离株的分子特征毒性及耐药性研究

目的 研究株蜡样芽胞杆菌SCY分离株的毒力和耐药性。方法 使用选择性培养基从银川市某餐厅鱼肉菜肴中培养分离获得蜡样芽胞杆菌SCY分离株,经23S rRNA基因PCR测序鉴定后,通过二代全基因组测序技术分析SCY分离株的基因组特征,进而分别采用免疫组化技术和药敏纸片实验研究SCY分离株的毒性和耐药性。结果 SCY分离株的基因组大小为5.82 Mb,编码基因个数为5 767,编码区总长度占全基因组的85.50%。SCY分离株基因组中共含有11个基因岛、16个CRISPR和5个前噬菌体;其中,SCY编码基因分别在PHI和VFDB毒力因子数据库中注释到了14个和62个毒力基因（Identiy>80,Evalue<0.05）,在CARD耐药数据库中注释到了215个耐药基因（Best Hit evalue<0.05）。小鼠肠道组织切片免疫组化结果表明,炎性因子IL-1β、CASP1和Nlrp3的表达水平发生差异显著性上调。22种细菌抗生素药敏纸片试验结果显示,SCY分离株对氯霉素、克林霉素等高度敏感,对四环素、头孢唑啉、头孢哌酮、氨苄青霉素表现为中介,对青霉素、苯唑西林、哌拉西林、杆菌肽、头孢他啶等10种抗生素表现为耐药,SCY分离株多重耐药情况较为严重。结论 本研究发现的SCY分离株属于肠毒株,携带多种肠毒素毒力基因,且具有典型的多重耐药特征。研究结果为揭示蜡样芽胞杆菌分离株的致病机制提供参考依据。     

The impact of oxygen availability on stress survival and radical formation of Bacillus cereus

Both the growth and stress survival of two model Bacillus cereus strains, ATCC 14579 and ATCC 10987, were tested in three different conditions varying in oxygen availability, i.e., aerobic, microaerobic and anaerobic conditions. Both B. cereus strains displayed highest growth rates and yields under aerobic conditions, whereas the microaerobic and anaerobic cultures showed similar reduced growth performances. The cells grown and exposed microaerobically and anaerobically were more resistant to heat and acid than cells that were cultured and exposed aerobically. On the other hand, the anaerobically grown cells were more sensitive to hydrogen peroxide compared to the (micro)aerobically grown cells. The increased heat- and acid-induced inactivation in aerobic conditions appeared to be associated with intracellular accumulation of excess hydroxyl and/or peroxynitrite radicals, as determined by flow cytometry in combination with the fluorescent reporter dye 3′-(p-hydroxyphenyl) fluorescein. This suggests that radical formation may contribute to inactivation of bacteria in the presence of oxygen, such as in aerobic and microaerobic conditions. No evidence was found for radical formation upon exposure to salt and hydrogen peroxide. The increased resistance to heat and acid in microaerobic and anaerobic conditions shows that oxygen availability should be taken into account when behavior of bacteria, such as B. cereus, in food industry related conditions is investigated, because oxygen availability may affect the efficiency of food preservation conditions.