吉林省肉及肉制品食源性致病菌检测与分析

目的 了解吉林省肉及肉制品食源性致病菌污染情况,分析主要的危险因素,为防控食源性疾病提供科学依据。方法 按照随机采样原则和食源性致病菌监测样本采样要求,对2011-2019年吉林省9个市的肉及肉制品采集5683株样品,依据GB/T 4789的标准方法,对沙门氏菌、金黄色葡萄球菌、单核细胞增生李斯特氏菌进行检测。结果 共检出阳性食源性致病菌314株, 总体检出率为5.53%, 其中沙门氏菌、金黄色葡萄球菌、单核细胞增生李斯特氏菌检出率依次为1.47%、5.51%、9.41%。在不同地点采集的标本中,农贸市场、酒店和超市检出率较高,集体食堂和学校周围小商铺均未检出。结论 吉林省各市不同场所的肉及肉制品存在不同程度的食源性致病菌污染,食品安全监管部门应根据污染程度实施有针对性的食品安全监督管理。     

单增李斯特菌毒力因子多重荧光PCR法 建立与应用

目的建立同时检测单增李斯特菌(Listeria monocytogenes)及其3种毒力因子的多重荧光PCR快速检测方法,并应用于日常食品的检测。方法根据单增李斯特菌溶血素基因hly A、内化素基因inl A和表面蛋白act A基因的保守序列,分别设计合成特异性引物和探针,优化多重荧光PCR反应体系。对该方法的特异性、敏感性和重复性进行评估。结果该法特异性强、敏感性高,对单增李斯特菌纯培养物的最低检出限410cfu/m L;重复性好,变异系数均小于2%。对84份食品检测结果与传统国标法相符,共检出单增李斯特菌4份,检出率为4.76%。多重荧光PCR检测方法耗时1 h,比传统方法节约2~5 d。4株单增李斯特菌分离株中2株同时含有inl A、act A、hly A 3种毒力基因,另2株为毒力基因act A缺失株,提示目前流行株并非同一来源。结论本研究建立的多重实时荧光PCR方法能同时对单增李斯特菌及其3种毒力因子进行快速检测,且灵敏度高、特异性好,为食源性疾病的病原学检测提供了快速可靠的方法。     

Edible coatings for enhancing microbial safety and extending shelf life of hard-boiled eggs

ABSTRACT:  Hard-boiled eggs were coated with chitosan–lysozyme (CL), whey protein isolate (WPI), or Bake sheen (BS), inoculated with Listeria monocytogenes or Salmonella enterica Ser. Enteritidis at 10⁴ CFU/g, and stored for 4 wk at 10 °C. Microbial populations were enumerated weekly. Two nonchallenge studies were also conducted with hard-boiled eggs: coated shell-on eggs packaged in plastic containers or peeled eggs coated and vacuum-packaged. Total plate counts (TPC), coliforms, yeasts, molds, weight loss, pH, and color of eggs were determined during the 10-wk storage at 10 °C. Uncoated eggs served as controls. All the coatings were not effective in inhibiting the growth of L. monocytogenes, but CL coating controlled the growth of S. Enteritidis. At the end of 4-wk storage, the numbers of S. enteritidis on CL-coated eggs were about 4-log₁₀ CFU/g less than that of the controls. Coatings reduced ( P < 0.05) the populations of coliforms and TPC, and completely inhibited mold growth during the 10-wk storage. Coatings also reduced ( P < 0.05) the weight loss of eggs, 4.1% to 4.8% on coated eggs compared with 7.5% in uncoated ones at the end of 10-wk storage. The pH of CL-coated eggs remained stable throughout the storage period, while the control eggs increased from 7.6 to 8.6. Color changes in CL- and WPI-coated eggshells were less ( P < 0.05) than those of BS-coated and the control. The CL coating effectively suppressed the numbers of TPC, coliforms, yeasts, and molds on peeled eggs during the 6-wk storage ( P < 0.05). The results suggest that CL coating on hard-boiled eggs can control the growth of S. Enteritidis and reduce undesirable changes in the interior quality of eggs.     

Recipes for antimicrobial wine marinades against Bacillus cereus, Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella enterica

ABSTRACT:  We have evaluated bactericidal activities against Bacillus cereus , Escherichia coli O157:H7, Listeria monocytogenes , and Salmonella enterica of several antimicrobial wine recipes, each consisting of red or white wine extracts of oregano leaves with added garlic juice and oregano oil. Dose-response plots were used to determine the percentage of the recipes that resulted in a 50% decrease in colony-forming units (CFU) at 60 min (BA₅₀). Studies designed to optimize antibacterial activities of the recipes demonstrated that several combinations of the naturally occurring plant-derived ingredients rapidly inactivated the above mentioned 4 foodborne pathogens. We also showed that (a) incubation temperature affected activities in the following order: 37 °C > 21 °C > 4 °C; (b) varying the initial bacterial concentrations from 10³ to 10⁴ to 10⁵ CFU/well did not significantly affect BA₅₀ values; (c) storage of 3 marinades up to 2 mo did not change their effectiveness against Salmonella enterica ; and (d) polyphenolic compounds isolated by chromatography from red wine exhibited exceptional activity at nanogram levels against 2 strains of Bacillus cereus . These observations suggest that antimicrobial wine formulations have the potential to improve the microbiological safety of foods.     

Antibacterial effects of natural tenderizing enzymes on different strains of Escherichia coli O157:H7 and Listeria monocytogenes on beef

This study determined the efficacy of actinidin and papain on reducing Listeria monocytogenes and three mixed strains of Escherichia coli O157:H7 populations on beef. The average reduction of E. coli O157:H7 was greater than that of L. monocytogenes and higher concentrations of either protease yielded greater reduction in bacterial populations. For instance, actinidin at 700 mg/ml significantly (p ≤ 0.05) reduced the population of L. monocytogenes by 1.49 log cfu/ml meat rinse after 3 h at 25 & 35 °C, and by 1.45 log cfu/ml rinse after 24 h at 5 °C, while the same actinidin concentration significantly reduced the populations of three mixed strains of E. coli O157:H7 by 1.81 log cfu/ml rinse after 3 h at 25 & 35 °C, and 1.94 log cfu/ml rinse after 24 h at 5 °C. These findings suggest that, in addition to improving the sensory attributes of beef, proteolytic enzymes can enhance meat safety when stored at suitable temperatures.     

食品中单核细胞增生李斯特氏菌的风险评估

微生物风险评估主要评估食品中的微生物性病原可能对人群引起的潜在危害,以指导风险管理者制定相应的管理措施。单核细胞增生李斯特氏菌是一种重要的食源性致病菌,由于该菌引发的疾病致死率较高,且其暴发常出现于工业加工食品中而引起了世界范围内的广泛关注。JEMRA及美国FDA/FSIS分别对即食食品中单核细胞增生李斯特氏菌进行了定量评估并发布了完整的评估报告。各国也有对本国特定食品中单核细胞增生李斯特氏菌进行评估的文献报道。但在数据不足的情况下,也可通过定性/半定量的风险分级工具对不同来源的风险进行分级,并确定优先性。由于各国人群消费模式、消费量的不同,以及食品制作和处理方法上的差异会对暴露评估的结果产生影响,从而影响每份食品风险的大小,因此,各国有必要根据本国的情况对特定食品中的单核细胞增生李斯特氏菌进行相应的评估。     

Effect of Cover Brine Type on the Quality of Meat from Herring Marinades

Effects of vinegar, oil, and sour cream brines on meat quality of 4 popular cold marinades from herring were investigated in the study. Cover brine type affected the composition and nutritive value of meat as well as the sensory and microbiological quality of marinated herring. Qualitative differences resulted from cover brine penetration into meat, and from diffusion of components from meat to vinegar brine. Compared to oil and sour cream, vinegar brine contributed to increased concentrations of salt and acetic acid, hardness, color brightness of marinades meat and to increased microbial contamination of meat. Furthermore, vinegar caused nitrogen losses to 15%, including valuable products of protein hydrolysis, enzymes, and total volatile bases. The rolling up of fillets reduced diffusion even by 50%. In turn, oil and sour cream were causing mainly a higher fat content and overall sensory evaluation of the marinades.     

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.     

食源性单核细胞增生李斯特菌四环素、红霉素耐药基因研究

研究了食源性单核细胞增生李斯特菌四环素、红霉素耐药基因的分布状况及和耐药表型的关系。采用微量肉汤稀释法对2005~2007年河北省疾病预防控制中心分离到的食源性单核细胞增生李斯特菌株进行四环素、红霉素药敏实验;应用PCR方法对实验菌株进行四环素耐药基因tet(M)、tet(S)、tet(L)、tet(K)、tet(B)、及与tet(M)基因关系密切的转座子Tn916、红霉素核糖体甲基化酶基因ermB、ermC、及与ermB基因关系密切的转座子Tn917检测,对阳性样本序列进行鉴定分析;应用血清学分型、脉冲场凝胶电泳(PFGE)、及脂肪酸聚类分析方法分析四环素耐药菌株之间的相关性,确定基因型和多态性。结果表明,91株单核细胞增生李斯特菌四环素敏感77株、耐药14株;红霉素敏感89株、耐药2株,其中包含1株菌同时交叉耐药四环素和红霉素;14株四环素耐药株中含tet(M)基因的13株,在13株tet(M)基因阳性菌中,tet(M)位于Tn916转座子上的9株;1株同时交叉耐药四环素、红霉素菌同时携带tet(S)、ermB基因;ermC基因、转座子Tn917均为阴性;四环素、红霉素敏感株中未检测到上述任何耐药基因。14株四环素耐药菌株血清型分布以1/2a型为主(n=12),部分菌株PFGE、脂肪酸分型完全一致。食源性单核细胞增生李斯特菌获得tet(M)基因是耐四环素的主要机制之一,具有水平传播耐药基因能力的接合型转座子Tn916与该菌四环素耐药播散有直接关系;ermB基因介导的核糖体靶位点改变存在食源性单核细胞增生李斯特菌红霉素耐药株中;PFGE基因型结合脂肪酸聚类分析能够用来分析菌株之间的相关性。     

基于CRISPR-Cas13的单增李斯特菌RNA快速检测方法的建立

为实现食品中单增李斯特菌污染的快速检测,本研究构建了一种基于CRISPR-Cas系统和Broccoli适配体的RNA均相检测技术。利用Cas 13与cr RNA锚定序列结合形成识别元件cr RNA-Cas13复合物,靶标RNA存在时可激活Cas 13的非特异性RNase活性,并利用点亮型RNA适配体Broccoli作为信号探针,监测cr RNA此-Cas13的活化状态。荧光值的变化与单增李斯特菌浓度存在线性关系,利用来检测单增李斯特菌。本研究所构建的检测可在30min内完成对于单增李斯特菌的的识别与检测,检出限为148CFU/m L,对细菌具有良好的检测特异性,可区分大肠杆菌、鼠伤寒沙门氏菌和蜡样芽孢杆菌。在牛奶模型中单增李斯特菌的加标回收率为95.15%～97.99%。该方法具有较好的灵敏度、特异性,可直接靶向检测致病菌RNA,无需逆转录、PCR扩增和核酸标记,简化了实验流程,对于实现食品中单增李斯特菌的现场检测及生物安全控制具有重要意义。