Response of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella typhimurium, and Staphylococcus aureus to the thermal stress occurring in model manufactures of Grana Padano cheese

The purpose of this research was to investigate the effect of temperature in the technology of production of Grana cheese against Escherichia coli O157:H7, Listeria monocytogenes, Salmonella Typhimurium, and Staphylococcus aureus. According to the technology of production, the cheese curds are cooked at 55°C and then cooled at room temperature (25°C). A curd-cooling model was developed to estimate the temperature variation across the curd during cooling, and the thermal stress was applied to the pathogens according to the model in model-scale productions of Grana cheese artificially contaminated with approximately 10⁴ cfu/mL of the selected pathogens. According to the numerical results, the initial temperature inside the cheese is kept at almost the initial value (above 50°C) for at least 4 h during cooling, whereas the crust of the curd cools rapidly to 30°C in the first hour. The best case was that of the core of the cheese where the high temperature was able to efficiently eliminate the contaminating pathogens. Moreover, the worst case was where the external ring of the curd in which a more rapid cooling allowed bacterial survival. Therefore, the thermal stress in the technology of production of Grana cheese can be only partially effective in the control of the selected pathogens. However, the whole technology of production includes other hurdles that can affect the survival of the pathogens and that need to be taken into account as a whole to evaluate the safety of Grana Padano cheese.     

肉中4种致病菌的PCR快速检测方法的建立

目的:建立一种能同时检测肉中金黄色葡萄球菌、志贺氏菌、沙门氏菌和单核增生李斯特菌的多重PCR检测方法。方法:根据金黄色葡萄球菌的耐热核酸酶基因(nuc)、沙门氏菌的侵袭蛋白基因(invA)、志贺氏菌的侵袭性质粒抗原基因(ipaH)和单核细胞增生性李斯特菌的内化素基因(inlA)设计引物,通过优化好的反应体系进行多重聚合酶链反应(PCR)扩增目的基因。结果:特异性实验结果表明4种菌均能在相应位置扩增出特异性条带。对污染4种菌的猪肉进行检测,确定出金黄色葡萄球菌和沙门氏菌的检出限是102CFU/mL,志贺氏菌和单核增生李斯特菌的检出限是101CFU/mL。结论:本实验建立的多重PCR方法比传统细菌检测方法更特异、快速、灵敏,适用于肉中金黄色葡萄球菌、志贺氏菌、沙门菌和单核增生李斯特菌的快速检测。     

The mitochondrial genome from the thermal dimorphic fungus Paracoccidioides brasiliensis

We present here the sequence of the mitochondrial DNA of the pathogenic thermodimorphic fungus Paracoccidioides brasiliensis, agent of an endemic disease in most South American countries. The sequenced genome has 71 334 bp and is organized as a circular molecule with two gaps of unknown size flanking the middle exon of the nad5 gene. We located genes coding for the three subunits of the ATP synthase (atp6, atp8 and atp9), the apocytochrome b (cob), three subunits of the cytochrome c oxidase enzyme complex (cox1, cox2 and cox3), seven subunits of the reduced nicotinamide adenine dinucleotide ubiquinone oxidoreductase (nad1, nad2, nad3, nad4, nad5, nad6 and nad4L) and the large (rnl) and small (rns) subunits of ribosomal RNA. Two maturases and a ribosomal protein (rms5) are located inside introns. Twenty-five tRNAs were identified with acceptors for all 20 amino acids. Seven polypurine/polypyrimidine tracts (140-240 bp) have been found in this genome. All genes are in the same orientation over the genome, while their order is closest to the mitochondrial genomes from Penicillium marneffei and Aspergillus nidulans.     

多重PCR技术在食源性病原细菌检测中的应用

食品中污染的病原细菌是引起食源性疾病的主要因素之一,快速检测食品中病原细菌是及时有效预防病原细菌传播及食物中毒的重要前提。多重PCR作为一种可同时检测多种病原细菌的方法应用日益广泛。本文介绍了多重PCR技术在食源性病原细菌检测中的应用现状及其多重PCR反应条件的优化,同时提出了存在的问题并对应用前景作了展望。     

非热杀菌技术在低温鸡肉制品致病菌控制中的应用研究进展

由于低温鸡肉制品具有较好的感官品质,能够保留鸡肉原有的营养,成为我国肉制品发展的一个趋势。低温鸡肉制品水分含量和pH值较高,在加工和贮藏过程中极易受到食源性致病菌的污染,影响产品的质量安全,甚至引发食源性疾病,因此采用合适的杀菌技术来解决致病菌的污染是低温鸡肉制品贮藏保鲜需要解决的关键问题。由于热杀菌技术不适合用于低温鸡肉制品的灭菌,非热杀菌技术的应用就显得尤其重要。目前,应用在低温鸡肉制品加工中的非热杀菌技术主要包括超高压、辐照、超声、低温等离子体、脉冲紫外线、脉冲电场和脉冲微波。本文主要综述以上7 种非热杀菌技术的杀菌机制以及在不同低温鸡肉制品加工应用时对致病菌的控制效果,为未来非热杀菌技术在低温肉制品加工中的应用提供参考。     

食源性致病菌污染估计中删失数据分析的研究进展

食源性致病菌污染水平的确定是开展微生物定量风险评估的重要前提,而删失数据的存在易造成对食品中致病菌整体污染水平的估计产生偏差.对检测过程中出现的删失数据进行分析研究已逐渐成为食源性致病菌定量建模工作的重要内容之一.本文对国内外相关研究进行综述,介绍了食源性致病菌污染检测中删失数据的分类,比较了替代法、参数估计法、非参数...     

多重PCR同时检测食品中4 种细菌与常见霉菌

建立一种同时检测食品中金黄色葡萄球菌、单核细胞增生李斯特氏菌、沙门氏菌、大肠杆菌O157:H7以及霉菌的多重聚合酶链式反应（polymerase chain reaction,PCR）检测方法。根据金黄色葡萄球菌的耐热核酸酶基因（nuc）、沙门氏菌的侵袭正调节蛋白基因（hilA）、大肠杆菌O157:H7鞭毛基因（flic）、单核细胞增生李斯特氏菌的毒力调控蛋白基因（prfA）及霉菌18S?rRNA基因V5区分别设计5?对特异性引物,并对PCR扩增反应体系和扩增条件进行优化,确定获得特异性良好的PCR扩增产物。而后在37?℃对人工污染致病菌的香肠、面包和豆腐进行增菌培养,5?种致病微生物在20?h内的检出限均可达到100?CFU/25?g。本实验建立的多重PCR检测方法适用于食品中4?种细菌与常见霉菌的同时检测,相较于传统检测方法,具有快速、简便、特异性高的优点。     

2016年广州市市售食品中食源性致病菌 监测结果分析

目的 了解广州市市售食品中食源性致病菌污染及分布情况, 发现危险因素。方法 2016年共采集6类共1058份食品样品, 对副溶血性弧菌、金黄色葡萄球菌、致泻大肠埃希氏菌、沙门氏菌、蜡样芽孢杆菌、单增李斯特氏菌等进行监测分析。结果 检出食源性致病菌164株, 总检出率为15.50%。食源性致病菌中副溶血性弧菌的检出率最高(40.93%), 其次为蜡样芽胞杆菌(9.29%)、金黄色葡萄球菌(2.38%)、单增李斯特氏菌(2.08%)、沙门氏菌(1.42%), 致泻大肠埃希氏菌检出率为0。不同食品类别中水产品检出率最高, 达到了43.52%, 其次为烘焙食品(13.64%)、小摊贩食品(10.62%)、生鸡肉(8.47%)、冷藏膳食(7.50%)和熟肉制品(6.82%), 不同食品食源性致病菌检出率差异有统计学意义。不同采样场所中采自餐饮店的食品食源性致病菌检出率最高(50.00%), 其次是农贸肉菜市场(17.70%)、超市(16.71%)和小摊贩(10.62%), 最低的是零售店(9.83%)。散装食品食源性致病菌检出率(15.85%)高于预包装食品的检出率(9.09%)。中心城区烘焙食品、生鸡肉致病菌检出率要高于周边区, 周边区域小摊贩食品致病菌检出率为零。结论 2016年广州市市售食品存在较高的食源性致病菌检出率, 对食品安全隐患较高的食品应加强监管, 预防和控制食源性疾病的发生。     

多重PCR检测4种常见食源性致病菌

目的 建立一种多重聚合酶链式反应法(multiplex polymerase chain reaction, MPCR)快速检测肉制品中金黄色葡萄球菌、沙门氏菌、志贺氏菌和单增李斯特氏菌的分析方法。方法 选取金黄色葡萄球菌nuc基因、沙门氏菌SipB基因、志贺氏菌ipaH基因、单增李斯特菌inlA基因作为目标基因, 设计4对PCR引物, 建立并优化多重PCR反应体系, 评价该体系的特异性和灵敏度, 并对人工污染的熟肉样品进行检测。结果 构建的多重PCR方法特异性强、灵敏度高, 人工污染熟肉匀浆中4种致病菌的检出限为103 CFU/mL。结论 构建的多重PCR检测方法能够快速、准确、高效地检测肉制品中金黄色葡萄球菌、沙门氏菌、志贺氏菌和单增李斯特氏菌, 为食源性疾病菌的快速检测提供参考依据。