高校周边速食品中病原微生物的检测与评价

由致病微生物引起的食源性疾病,已成为国内主要的食品安全问题之一。为了解天津某高校周边市场即食食品的微生物污染状况,本课题采取随机抽样方法采集天津某高校周边小吃街等场所的主食类、卤制品类、小吃类、凉拌菜类、糕点类、沙拉寿司水果、饮品类等7类别共计50件食品,根据GB/T4789和国家卫生标准,对7类食品中的菌落总数和大肠菌群及主要致病性微生物,包括金黄色葡萄球菌、沙门氏菌、志贺氏菌等进行全程调查与评价。调查结果显示,大肠菌群,金黄色葡萄球菌和沙门氏菌合格率较低,分别是74.98%,62.17%和51.72%。在各类食品中,合格率分别是饮品(75%)>糕点(66.67%)>小吃类(57.89%)>主食类(56.25%)>沙拉寿司(50%)>卤制品(25%)>凉拌菜(0%),在7类食品中,除了饮品以外,都检测出不同程度的致病菌微生物,其中以卤制熟食品和凉拌菜类的合格率最低,分别是25%和0%。主食类、卤制品、小吃类、凉拌菜类3种致病菌均有检出,糕点类只检出沙门氏菌,沙拉寿司水果检出金黄色葡萄球菌和沙门氏菌。研究结果显示,高校周边市场即食食品微生物污染情况严重,致病菌检出率较高,相关部门应该加强对学生的食品安全教育和周边市场的卫生监督,防止食源性疾病发生。     

Molecular Mechanisms of Host Cytoskeletal Rearrangements by Shigella Invasins

Pathogen-induced reorganization of the host cell cytoskeleton is a common strategy utilized in host cell invasion by many facultative intracellular bacteria, such as Shigella, Listeria, enteroinvasive E. coli and Salmonella. Shigella is an enteroinvasive intracellular pathogen that preferentially infects human epithelial cells and causes bacillary dysentery. Invasion of Shigella into intestinal epithelial cells requires extensive remodeling of the actin cytoskeleton with the aid of pathogenic effector proteins injected into the host cell by the activity of the type III secretion system. These so-called Shigella invasins, including IpaA, IpaC, IpgB1, IpgB2 and IpgD, modulate the actin-regulatory system in a concerted manner to guarantee efficient entry of the bacteria into host cells.     

志贺氏菌噬菌体空间杀菌效力的研究

将福氏志贺氏菌噬菌体SF-2A、痢疾志贺氏菌噬菌体SD-11及宋氏志贺氏菌噬菌体SS-92以等比例混合后进行气雾喷洒分析其在空间的杀菌效力,以期在食品生产和加工中控制食源性志贺氏菌的污染。首先在空间分析不同噬菌体气雾喷洒后的杀菌活性和最佳作用浓度,其次将福氏、痢疾及宋氏志贺氏菌以等量进行混合,以3×105cfu/mL的终浓度进行人工喷洒污染,并以3×108cfu/mL的噬菌体混合物进行气雾喷洒,分别在喷洒后1 h,2 h和3 h检测宿主菌的数量。结果显示,噬菌体SF-2A、SD-11及SS-92均能在空间有效灭活其相应宿主菌,其混合溶液作用3 h后已检测不到志贺氏菌。由此可见,混合噬菌体能够有效控制志贺氏菌的污染,这为食品生产及加工中环境中的病原菌的污染控制提供了新途径。     

Assessment of the toxicity of two Aconitum herbal medicines by microcalorimetry and chemometrics

In this study, a non-invasive and non-destructive microcalorimetric method was applied to assess the toxic effects of two Aconitum herbal medicines, Radix Aconitum Kusnezoffii and Radix A. carmichaeli, on the multiplying and non-multiplying metabolism of Shigella dysenteriae. Some crucial information, such as the thermogenic curves and thermo-kinetic parameters including growth rate constant k, inhibitory ratio I and half-inhibitory concentration IC₅₀, of S. dysenteriae growth affected by the two Aconitum species was obtained. Coupled with chemometric analysis on this information, it could be found that Radix A. Kusnezoffii and Radix A. carmichaeli could be differentiated according to their toxic effects; Radix A. Kusnezoffii with IC₅₀ of 35.2 mg/mL had stronger toxic effect than Radix A. carmichaeli with IC₅₀ of 49.6 mg/mL. This present study shows the effectiveness of combining microcalorimetry and chemometrics for assessing the toxic effects of herbal medicines and other food matrices.     

副干酪乳杆菌M5-L抑制宋内志贺氏菌黏附作用影响因素的研究

研究副干酪乳杆菌M5-L抑制宋内志贺氏菌(S.sonnei)对HT-29细胞黏附的作用及其影响抑制黏附作用的表面分子分析。结果表明M5-L能够显著的抑制S.sonnei的黏附作用,在排除、竞争和取代黏附实验中,抑制率分别为55%、33%和30%,差异显著(p<0.05),高碘酸钠处理后,M5-L的抑制黏附作用没有变化,表明碳水化合物并没有参与到黏附抑制过程中。而经过5mol/L LiCl处理后,M5-L的抑制黏附作用下降,说明S-层蛋白参与了M5-L抑制S.sonnei对HT-29细胞的黏附作用。此外,M5-L的S-层蛋白能够显著抑制S.sonnei的黏附作用。     

靶序列富集多重PCR结合DHPLC同时检测5种食源性致病菌

目的建立同时检测沙门氏菌、单核细胞增生李斯特氏菌、大肠埃希氏菌O157、金黄色葡萄球菌和志贺氏菌5种食源性致病菌的简便、灵敏的高通量方法。方法根据GenBank公布的5种致病菌的特异性靶基因序列,设计5对特异性引物,与通用引物连接构成复合引物。经过反应条件的优化,建立了5种致病菌的靶序列富集多重PCR(target enriched multiplex PCR,Tem-PCR)扩增方法,扩增产物采用变性高效液相色谱(denaturing high-performance liquid chromatography,DHPLC)技术进行检测。结果采用Tem-PCR-DHPLC方法,检测48株试验菌株,未发现非特异性结果。对5种致病菌检测灵敏度,除金黄色葡萄球菌为620 cfu/mL外,其余都小于100 cfu/mL。对4种样品的检测结果与国家标准方法相符合。结论 Tem-PCR-DHPLC检测方法,相比传统多重PCR方法,能有效地解决引物扩增效率不均衡的问题,不需优化引物浓度,方法特异性强,灵敏度高,具有较好的实用性。     

Inactivation of Shigella boydii 18 IDPH and Listeria monocytogenes Scott A with power ultrasound at different acoustic energy densities and temperatures

ABSTRACT:  The effect of acoustic energy density (AED) on inactivation of Shigella boydii 18 IDPH and Listeria monocytogenes Scott A in a cell suspension was studied at sublethal temperatures and at AEDs of 0.49, 0.85, and 1.43 W/mL. The effect of temperature on ultrasonic inactivation of L. monocytogenes Scott A at 35, 50, and 65 °C was examined at an AED of 1.43 W/mL. Increasing AED increased the rate of inactivation for both S. boydii and L. monocytogenes . The destruction of S. boydii and L. monocytogenes followed 1st order kinetics in a 20-min treatment, except for S. boydii inactivation at 1.43 W/mL where a tailing effect was observed after 15 min. At sublethal temperatures, the D-values of S. boydii were 8.8, 4.3, and 2.5 min for AEDs of 0.49, 0.85, and 1.43 W/mL, whereas those for L. monocytogenes at the 3 AED levels were 31.5, 13.5, and 7.3 min, respectively. Ultrasonic treatment of L. monocytogenes at 35 and 50 °C enhanced inactivation. However, at 65 °C, application of ultrasound did not result in additional inactivation compared to thermal treatment alone at the same temperature. With the experimental conditions and the ultrasound system used in this study, an upper temperature limit for thermosonication was evident above which no added killing due to ultrasound was observed.     

抗志贺氏菌IgY的提纯及建立间接ELISA检测志贺氏菌

采用水稀释法提纯抗志贺氏菌IgY,检测10mg/mL纯化抗志贺氏菌IgY的效价为1∶320,并以此IgY为基础建立间接ELISA检测志贺氏菌,测定志贺氏菌纯培养液的检出限为105～106cfu/mL。对蜡样芽孢杆菌等10株不同菌株的检测结果表明,该方法对志贺氏菌有明显的检测特异性,对所测定的其它菌株无交叉反应。     

二重PCR法快速检测3种食源性致病菌

目的建立快速检测食品中金黄色葡萄球菌(Staphylococcus aureus)、沙门氏菌属(Salmonella spp.)及志贺氏菌属(Shigella spp.)的二重PCR方法。方法分别针对金黄色葡萄球菌.femA和nuc、沙门氏菌属invA和hilA及志贺氏菌属ipaB和ipaH设计6对特异性引物,检测每对引物的特异性及灵敏度,组合优化各自二重PCR反应体系。结果初步建立了针对这3类致病菌的二重PCR检测方法,整个检测过程不超过18 h,检测灵敏度均可达10 pg DNA/reaction。结论所建立的针对3类致病菌的二重PCR检测方法具有灵敏度高、特异性强和方便高效等优点,具有良好的市场应用前景。     

肠道杆菌的扫描电镜观察

我们用扫描电镜观察肠道杆菌种(Enterobacteriaceae)中的四种细菌,大肠埃希氏杆菌(Escherichiacoli),普通变形杆菌(Proteus vulgaris),伤寒沙门氏菌(Salmonella typhi)和福氏志贺氏菌(Shigella Flexneri)。未处理的菌落标本,菌落表面都可形成一层厚薄不同的表膜(Surface film)。四种细菌菌落的表膜,形状不一,千姿百态。适当处理后的菌落标本,则可显示菌体的本来面目。四种细菌菌体在菌落表面的分布和排列,也是各不相同,千差万别。本文根据扫描电镜的观察,对四种不同菌落进行了讨论。