单增李斯特菌的生长特性及其在冷藏牛奶中的预测模型

通过测定单增李斯特菌菌株在不同培养条件下菌液的OD600值,绘制生长曲线,分析培养基成分、温度、p H、Na Cl浓度、接种量以及转速对单增李斯特菌生长的影响,并选取Gompertz方程、Logistic方程和Hill方程建立单增李斯特菌在冷藏牛奶中的生长模型。结果表明:菌株在TSB-YE和MRS培养基中生长良好;能够耐受0.5%³%的Na Cl浓度范围;最适生长温度为37℃,最适生长p H为8,对氧气的需求量表现不明显。菌株可在4℃冷藏的全脂牛奶中缓慢生长,达到稳定期的菌数有2 lg CFU/m L的增长,Gompertz和Logistic方程模型可用于预测其生长情况。      

Evaluation of Listeria innocua as a suitable indicator for replacing Listeria monocytogenes during ripening of Camembert cheese

The suitability of Listeria innocua for use as an indicator for replacing Listeria monocytogenes during the cheese-making and ripening of Camembert cheese was evaluated. Pasteurized whole milk inoculated with either L. innocua or L. monocytogenes was used to make Camembert cheese, which were ripened in three stages. All cheese was ripened in three stages: room temperature (∼20 °C) and relative humidity of 60% for 36 h; 12 °C and relative humidity of 93% for 2 weeks; and 7 °C and relative humidity of 85% for 3 weeks. Results showed that population values of L. innocua and L. monocytogenes on day 1 were 7.16 and 6.11 log₁₀ CFU/g, respectively, which declined to 6.54 and 5.45 log₁₀ CFU/g, respectively, during subsequent 20 days. Thereafter, L. innocua and L. monocytogenes populations increased to 7.38 and 6.06 log₁₀ CFU/g on day 35 of ripening, respectively. During ripening, surface and interior of cheeses were analysed for populations of L. innocua and L. monocytogenes , respectively. The data were collected on day 1, 5, 10, 15, 20, 25, 30, and 35 of ripening. Generally, the growth of L. innocua and L. monocytogenes is faster in surface than in centre. Top centre, bottom centre and bottom surface locations had similar population values during ripening. There were no significant differences ( P  > 0.05) between batch and section of cheese. The ripening time and locations had significant effect ( P  < 0.05) on the survival and growth of L. innocua and L. monocytogenes . The trends of survival and growth of L. innocua and L. monocytogenes were similar. These results indicated that L. innocua can be considered as an indicator for L. monocytogenes during ripening of Camembert cheese.     

单增李斯特菌在牛奶中生长曲线的拟合和预测模型的建立

通过生长动力学曲线拟合,建立牛奶中单增李斯特菌的生长预测模型。比较了5、15、25和35℃条件下Gompertz方程、Logistic方程和Hill方程的拟合情况,确定Gompertz方程为单增李斯特菌在牛奶中最优的初级模型。采用Belehradek方程描述温度对最大生长速率和延滞时间的影响,结果表明它们之间成良好的线性关系(R2分别为0.9721和0.9747),并建立了相应的二级模型。对20℃条件下由预测模型构建的微生物生长动力学方程进行了验证,同时对模型的可靠性进行了分析,结果表明预测值和真实值的残差均小于0.2,模型的偏差度为1.04,准确度为1.13,表明建立的模型有效且可靠。在已知初始菌数的情况下,可以根据模型快速预测5~35℃条件下不同贮藏时间点单增李斯特菌在牛奶中的生长情况。      

Occurrence and Antimicrobial Susceptibility of Listeria monocytogenes Isolated from Brined White Cheese in Jordan

Listeria monocytogenes is a serious foodborne pathogen that has been isolated from different dairy food products. Several foodborne outbreaks of listeriosis have been associated with consumption of cheese. The aims of this study were to determine the occurrence of L. monocytogenes and Listeria spp. in brined white cheese (BWC) sold in Jordan, and to determine the susceptibility of isolated L. monocytogenes to antimicrobials. Three hundred and fifty samples of 5 different types of BWC (akkawi, boiled, halloumi, pasteurized, and shellal) were collected from a local market in Jordan. The ISO (11290-1) procedure was followed for isolation and identification of Listeria spp. from cheese samples and a polymerase chain reaction (PCR) technique was used for confirmation of L. monocytogenes isolates. The VITEK2 automated system was used for testing antimicrobial susceptibility of L. monocytogenes isolates. The overall prevalence of Listeria spp. in cheese sample was 27.1%. L. monocytogenes was isolated from 39 (11.1%) samples. Other isolated species were L. grayi (6.9%), L. innocua (2%), L. ivanovii (4%), L. seeligeri (2%), and L. welshimeri (0.3%). The pH values and salt concentrations of L. monocytogenes positive cheese samples ranged from 5.10 to 6.32 and 5.64 to 13.16, respectively. L. monocytogenes isolates were sensitive or intermediate susceptible to imipenem, gentamicin, linezolid, teicoplanin, vancomycin, fusidic acid, trimethoprim/sulfamethoxazole, benzylpenicillin, ciprofloxacin, erythromycin, tetracycline, and rifampicin, but resistant to fosfomycin, oxacillin, and clindamycin.     

食品中单核细胞增生李斯特氏菌调查及毒力研究

为了解和掌握各类食品中单核细胞增生李斯特氏菌（Lm)污染状况及菌株的毒力情况,采集5类265件食品,菌株鉴定应用API细胞鉴定系统;毒力测定采用溶血实验、多聚酶链反应（PCR）以及小鼠致病力实验的方法进行,结果显示Lm检出率为4.90%;其中从生肉、熟肉、灌汤和速冻食品中的检出率分别为15.00%、2.48%、9.52%;从熟肉和速冻食品中还检出其它李斯特氏菌,检出率分别为4.96%、5.92%;乳制品和水产品中未检出Lm。13株Lm毒力测定显示,溶血实验与小鼠的致病力和PCR测定的溶血素基因无必然联系,而溶血素基因与内化素基因阳性结果相同,Lm对多种抗生素敏感,其中对氨氨苄青霉素、头孢唑啉和环丙沙星敏感率为100%。     

云南省牛乳中单核细胞增生李斯特氏菌的分布特征、耐药性及毒力研究

目的 分析云南省牛乳中单核细胞增生李斯特氏菌(Listeria monocytogenes)的分布特征、耐药性及毒力现状.方法 采集云南省主要奶产区牛乳样本,按照GB 4789.30—2016方法分离单核细胞增生李斯特氏菌疑似菌株,采用飞行时间质谱技术结合16S rDNA测序对疑似菌株进行鉴定;采用微量肉汤稀释法对分离...     

Identification of Listeria innocua surrogates for Listeria monocytogenes in hamburger patties

ABSTRACT:  Listeria innocua M1 has been used by many researchers as a nonpathogenic thermal processing surrogate for Listeria monocytogenes . However, L. innocua M1 has been criticized because its thermal survivability characteristics are not as closely parallel to L. monocytogenes as some would like in a variety of foods and processing conditions. The present study was conducted to compare multiple L. innocua and L. monocytogenes strains to validate L. innocua M1 as the ideal surrogate under high-temperature thermal processing conditions for L. monocytogenes . The D - and z -values of L. innocua M1, L. innocua strain SLCC 5639 serotype (6a), SLCC 5640 (6b), SLCC 2745 (4ab), and L. monocytogenes F4243 (4b) were calculated for raw hamburger patties. Hamburger patties were inoculated with 10^7–8 CFU/g of L. monocytogenes or L. innocua . Samples were heat treated at 4 temperatures (62.5 to 70 °C). At each temperature, the decimal reduction time ( D -value) was obtained by linear regression of survival curves. The D - and z -values were determined for each bacterium. The D -values of L. innocua and L. monocytogenes serotypes ranged from 3.17 to 0.13 min at 62.5 to 70 °C, and the z -values of L. innocua and L. monocytogenes were 7.44 to 7.73 °C. Two of the 4 L. innocua serotypes used in this experiment have the potential for use as surrogates in hamburger meat with varying margins of safety. L. innocua M1 should serve as the primary nonpathogenic surrogate with the greatest margin of safety in verifying a new thermal process to destroy L. monocytogenes .     

单核细胞增生李斯特菌菌株分型与其致病潜力基因相关性研究进展

单核细胞增生李斯特菌是一种常见的食源性致病菌,该菌分型繁多。不同分型的单核细胞增生李斯特菌致病潜力不同,这与菌株所携带的抗性基因和毒力基因不同有关。本文综述了不同分型单核细胞增生李斯特菌与抗性基因和毒力基因的关系,结果发现与该菌抗性相关的基因,如热抗性、耐冷性、酸抗性、耐高渗、耐干燥、耐金属和杀菌剂及参与应激蛋白表达调控的基因较多,它们与分型之间的关系暂无明确相关性结论,但也发现应激生存岛（stress survival island,SSI）-1仅存在于谱系I和谱系II部分菌株中,SSI-2目前仅被发现在ST121菌株中携带。从功能毒力基因和李斯特菌毒力基因岛（Listeria pathogenicity islands,LIPI）两方面分述了毒力基因,LIPI-3和LIPI-4主要存在于谱系I菌株中。此外,ST5、ST8、ST9和ST121菌株的inlA基因中出现提前终止密码子,使得菌株的毒力降低。研究不同分型单核细胞增生李斯特菌致病潜力基因的差异有助于单核细胞增生李斯特菌的预警预测、防控,并为不同分型菌株致病机制的深入研究提供参考。     

环介导恒温扩增方法快速检测乳中单增李斯特菌

建立了一种快速检测灭菌乳中单增李斯特菌的环介导恒温扩增(Loop-Mediated Isothermal Amplification,LAMP)方法。以hlyA基因作为靶基因,对人工污染乳中单增李斯特菌进行了LAMP方法的灵敏度试验,同时与PCR方法进行比较。并对单增李斯特菌和7种其他乳中常见致病菌进行了LAMP检测,以验证该方法的特异性。结果表明,LAMP检测单增李斯特菌的特异性强,检出限为42 mL-1,其灵敏度比普通PCR高10倍。并且检测时间比PCR更短,在1.5 h内即可完成扩增反应。此方法快速、特异、简单、灵敏,具有较高的推广价值。     

生鲜乳中单核细胞增生李斯特菌的研究进展

单核细胞增生李斯特菌是一种重要的人畜共患食源性致病菌,普遍存在于生鲜乳中,严重威胁生鲜乳质量安全。本文就单核细胞增生李斯特菌的生物学特征和致病性、检测方法和生鲜乳中单核细胞增生李斯特菌的检出率的研究进展以及对生鲜乳中单核细胞增生李斯特菌的防控措施进行综述,为生鲜乳质量安全风险评估提供一定的理论基础。     

单核细胞性李斯特菌基因dal的敲除及其生物学特性初步分析

在单核细胞性李斯特菌(Listeria monocytogenes)野生株EGDe act A及inl B双基因缺失株(EGDeΔact AΔinl B)的基础上,利用同源重组的方法进一步构建了缺失营养基因dal的菌株(EGDe Δact AΔinl BΔdal),并对该缺失菌株生长状态、毒力基因表达水平、生物被膜的形成量及细胞侵袭等方面作进一步分析。结果显示,37℃摇床培养6 h后,缺失株的菌浓度显著低于EGDe Δact AΔinl B(P0.001),培养基中补充D-丙氨酸的缺失株生长速率与亲本株相比无显著差异;实时荧光定量聚合酶链式反应结果显示,缺失株的sig B基因表达水平变化最明显(P0.01),约下调90%;缺失株生物被膜形成量显著增加(P0.05),培养基补充D-丙氨酸后缺失株生物被膜的生成量与亲本株相比无差异;对Coca-2细胞的侵袭无影响,表明该基因对细菌生长能力及生物被膜形成具有重要的调控作用,并不影响菌株对细胞的侵袭力。此缺失株的构建为进一步研究基因dal的功能提供了理论支持。     

Growth of Listeria monocytogenes in refrigerated ready-to-eat foods in Japan

The ability of L. monocytogenes to grow in a series of Japanese ready-to-eat (RTE) foods, including boiled baby sardine and Japanese pickle, was tested at two different refrigeration temperatures. In RTE foods in which L. monocytogenes can grow, growth was significantly higher at 10°C than that at 4°C during their shelf lives and growth patterns varied extensively among the different types of foods. However, growth did not occur at 4°C within the shelf life of certain RTE foods, such as broiled squid. The patterns of growth were varied extensively with different sample types. These results suggest that some types of traditional Japanese RTE foods stored at 10°C may be potential sources of listeriosis. To reduce the risk of food-borne listeriosis, studies to determine the contamination levels in RTE foods and the effects of storage temperature on their shelf lives are needed.     

The occurrence of Listeria monocytogenes in cheese from a manufacturer associated with a case of listeriosis

A case of listeriosis was associated with the consumption of a soft cheese produced in England. Goats cheese and other products from the same food manufacturer were examined for the presence of Listeria over the following 11 months. Listeria monocytogenes was isolated from 16 of 25 cheese samples on retail sale, 12 of 24 cheese samples obtained directly from the factory, and from shelving within the plant. Phage-typing of 68 isolates of L. monocytogenes from cheese samples and the factory showed that 66 (97%) were indistinguishable from the strain isolated from the patient's cerebrospinal fluid and stool. L. monocytogenes was not isolated from seven goats milk or two yoghurt samples. Listeria innocua was isolated from 10 cheese samples, two of which contained no other species of Listeria. Levels of L. monocytogenes shortly after production were low (<10/g), but were higher (10⁵–10⁷ cfu/g) in six of the 16 cheese samples obtained from retail outlets. Multiplication of L. monocytogenes was demonstrated in cheeses contaminated at the factory and held at 4°C in the laboratory.     

Listeria monocytogenes: A Target for Bacteriophage Biocontrol

Listeria monocytogenes is a growing concern in the food industry as it is the causative agent of human listeriosis. There are many research articles concerning the growth, survival, and diversity of L. monocytogenes strains isolated from food‐related sources, elucidating the difficulty in controlling these bacteria in a food‐processing facility. Bacteriophage biocontrol of L. monocytogenes strains was introduced in 2006, through the first commercial bacteriophage product targeting L. monocytogenes ListShield^TM. This review focuses on the use of bacteriophage biocontrol to target L. monocytogenes in the food industry, specifically direct application of the bacteriophages to food products. In addition, we discuss characteristics of these bacteria that will have a significant influence on the effective treatment of bacteriophages such as genetic diversity between strains prevalent in one facility. There are many positive results of phage treatments targeting L. monocytogenes in food; however, success of in vitro studies might not be reproducible in practice. Future studies should focus on creating experimental design that will imitate the conditions found in the food industry, such as a stressed state of the targeted bacteria. In situ evaluation of bacteriophage treatment of L. monocytogenes will also be necessary because the presence of these bacteria in a processing facility can vary greatly regarding genetic diversity. The potential use of phages in the food‐processing facility as a biosanitizer for L. monocytogenes, as well as the use of lysins to target these bacteria should also be explored. Despite the exciting research avenues that have to be explored, current research shows that biocontrol of L. monocytogenes is feasible and has potential to positively impact the food industry.     

The occurrence,growth, and biocontrol of Listeria monocytogenes in fresh and surface-ripened soft and semisoft cheeses

Listeria monocytogenes continues to pose a food safety risk in ready-to-eat foods, including fresh and soft/semisoft cheeses. Despite L. monocytogenes being detected regularly along the cheese production continuum, variations in cheese style and intrinsic/extrinsic factors throughout the production process (e.g., pH, water activity, and temperature) affect the potential for L. monocytogenes survival and growth. As novel preservation strategies against the growth of L. monocytogenes in susceptible cheeses, researchers have investigated the use of various biocontrol strategies, including bacteriocins and bacteriocin-producing cultures, bacteriophages, and competition with native microbiota. Bacteriocins produced by lactic acid bacteria (LAB) are of particular interest to the dairy industry since they are often effective against Gram-positive organisms such as L. monocytogenes, and because many LAB are granted Generally Regarded as Safe (GRAS) status by global food safety authorities. Similarly, bacteriophages are also considered a safe form of biocontrol since they have high specificity for their target bacterium. Both bacteriocins and bacteriophages have shown success in reducing L. monocytogenes populations in cheeses in the short term, but regrowth of surviving cells can commonly occur in the finished cheeses. Competition with native microbiota, not mediated by bacteriocin production, has also shown potential to inhibit the growth of L. monocytogenes in cheeses, but the mechanisms are still unclear. Here, we have reviewed the current knowledge on the growth of L. monocytogenes in fresh and surface-ripened soft and semisoft cheeses, as well as the various methods used for biocontrol of this common foodborne pathogen.     

单核细胞增生李斯特菌的毒力因子

目的单核细胞增生李斯特菌(Listeria monocytogenes,Lm)属于革兰阳性无芽孢杆菌李斯特菌属,主要通过食物传播。Lm的致病性与毒力因子密切相关,研究其毒力因子对认识致病机理有着重要意义。方法对Lm重要的毒力因子(溶血素、磷脂酶、内化素、肌动蛋白、P60蛋白等)进行综述。结果溶血素是一个多功能的毒力因子,对于Lm逃离吞噬细胞囊是必需的。Lm能产生两种磷脂酶C:磷脂酰肌醇磷脂酶C和磷脂酰胆碱磷脂酶C,协助细菌的细胞内复制。肌动蛋白使得Lm在宿主细胞间能够扩散。内化素与Lm的侵袭力有关。P60蛋白是Lm的主要免疫原性抗原。结论Lm毒力因子研究的深入对李斯特菌病的防治将带来深远的影响。     

A comparison of gene expression of Listeria monocytogenes in vitro and in the soft cheese Crescenza

This study focuses on the expression analysis of virulence and stress response genes (plcA, hly, iap and sigB) in 11 different strains of Listeria monocytogenes. The first step of this study considered an in vitro analysis in synthetic laboratory medium. According to statistical analysis, significant differences emerged for genes sigB and plcA. Subsequently, a soft cheese at two temperatures (4 and 12 °C) and for different times (24 and 48 h) was evaluated. Significant expression differences emerged for the condition at 12 °C after 48 h. No association could be observed between gene expression profile and origin of the different strains.     

单核细胞增生李斯特氏菌的PCR检测方法

研究比较了裂解法、热煮沸法、试验盒法提取单增李斯特氏菌DNA的效果 ,认为Promega试剂盒法提取的DNA ,得率高、纯度好。同时根据单增李斯特氏菌的毒力相关基因的 5对引物 ,进行了引物的特异性研究 ,结果发现inlA引物、inlB引物、plcA引物特异性好 ,可以用于食品中单增李斯特氏菌的检测     

单核细胞增生性李斯特菌自溶素研究进展

自溶素是由细菌产生的可以降解细菌细胞壁的肽聚糖水解酶,参与细菌的分裂、细胞壁更新、细菌自溶等生理过程。自溶素也是细菌的重要毒力因子之一,参与细菌的黏附与侵袭,与细菌的致病性密切相关。本文对单核细胞增生性李斯特菌的胞壁水解酶p60、酰胺酶Ami、自溶素IspC、自溶素Auto与胞壁水解酶MurA等自溶素的蛋白结构、生理功能及致病性进行了简要介绍。