噬菌体对虾仁中单核细胞增生李斯特菌的抑制效果

以虾仁为研究对象,通过人为模拟在虾仁中接种单核细胞增生李斯特菌后,再用噬菌体ListexTMP100在常温(20℃)、冷藏(4℃、0℃)和冷冻(-18℃)环境下进行处理,分析虾仁中该菌的抑制效果,单核细胞增生李斯特菌采用平板计数法进行计数。结果表明:噬菌体浓度大于2×107PFU/mL时,能有效杀灭虾仁中单核细胞增生李斯特菌(P<0.05);该浓度在20℃下作用虾仁1 h,便能降低1.50 Log10CFU/g(死亡率为99%以上)的单核细胞增生李斯特菌;在0℃和4℃,作用24 h下,单核细胞增生李斯特菌的死亡率达到99.9%;在-18℃下贮藏30d,单核细胞增生李斯特菌在第1天下降了1.38Log10CFU/g。这些结果表明,噬菌体ListexTMP100对虾仁中的单核细胞增生李斯特菌具有明显的抑制效果,可作为一种理想的生物杀菌剂用于水产食品中。     

基于FSO值对生熟食品交叉污染中单增李斯特菌的危害识别

为定性分析生熟食品中单增李斯特菌在不同案板材质上的交叉污染,选定木制、塑料及不锈钢案板为材质研究对象,设置4种常见场景,并测定各案板材质不同场景下卤猪舌、案板与黄瓜中单增李斯特菌的污染水平(lg CFU/g)。同时,选择一级Baranyi模型修改式,二级主参数模型对黄瓜中单增李斯特菌的生长进行模拟。此外,根据食品安全目标(FSO)计算整个交叉污染过程的污染水平,并以即食食品中单增李斯特菌的FSO值2(lg CFU/g)为标准,与12种交叉污染情景(4种场景×3种材质)比较,并进行风险等级排序。结果表明:木质案板场景1(S1w)、塑料案板场景1(S1p)和不锈钢案板场景1(S1s)风险等级最高;场景2、3和4下,木质案板风险等级分别为2、3和3,高出塑料和不锈钢案板(3、4和4)一个风险等级。本研究表明对案板不进行任何清洗处理最易引发交叉污染,木质案板相较于其他2种案板材质的风险较大。     

酵母菌促进单核细胞增生李斯特氏菌的生长

为了缩短单核细胞增生李斯特氏菌(单增李斯特氏菌)的前增菌时间,提升检测效率,本研究将酵母菌(ATCC9763)作为一种生长促进剂加入到单核细胞增生李斯特氏菌的前增菌培养基(LB1)中,结果表明添加适量的酵母菌可以使单增生李斯特氏菌的生长速度提升近100%,并且这种促进作用与培养过程中酵母菌的接种量、培养基的溶氧量以及培养方式密切相关。根据酵母菌的生长特性分析,这种促进作用产生的原因很有可能是由于酵母菌在生长过程中发酵分解了培养基中的糖类等营养物质,改善了单增李斯特氏菌的营养条件,从而提高了单增李斯特氏菌的繁殖速度。本研究为缩短单增李斯特氏菌检测的富集培养时间,提高检测效率奠定了很好的基础,同时也侧面提示我们要关注发酵类食品遭受单增李斯特氏菌污染的食品安全风险。     

Utilization of Zein Coating and Sorbic Acid to Reduce Listeria monocytogenes Growth on Cooked Sweet Corn

Several proteins, lipids and waxes were tested as edible coatings on sweet corn. Only zein, a natural constituent of corn, gave a continuous adhesive and stable coating with satisfactory sensory properties. After 8 days at 10 °C, the population of L. monocytogenes was 10‐fold lower on coated sweet corn than on non‐coated sweet corn indicating a barrier effect of zein coating. Sorbic acid was incorporated in the coating at a concentration required to inhibit L. monocytogenes growth (approximately 1 mg sorbic acid/g of sweet corn). The inhibitory concentration was the same for both coated and non‐coated sweet corn. Zein coating therefore did not improve the preservative effect of sorbic acid.     

低温条件下即食虾中单增李斯特菌与副溶血性弧菌共存分子预测模型的建立

本研究旨在应用实时荧光定量聚合酶链式反应（quantitative real-time polymerase chain reaction,qPCR）技术描述即食虾中的单增李斯特菌与副溶血性弧菌的生长行为,构建混菌模式下的分子预测模型。将单增李斯特菌与副溶血性弧菌等量（（5.0±0.5）（lg（CFU/g）））混合接种于即食虾中,置于低温环境（4 ℃和10 ℃）下培养,并利用qPCR定量检测单增李斯特菌与副溶血性弧菌数量的动态变化。运用生长模型（修正Gompertz、Logistic、Baranyi）和失活模型（Log-linear、Weibull）分别拟合两株菌的生长和失活趋势。结果表明：低温条件下,修正Gompertz、Logistic和Baranyi模型均可成功拟合单增李斯特菌的生长曲线,其决定系数（R2）均大于0.98。对于副溶血性弧菌,在4 ℃条件下,Log-linear和Weibull模型能够清晰地描述其失活情况,R2分别为0.950和0.945;而10 ℃条件下,2 个失活模型均难以描述其行为变化,R2仅为0.784和0.775。本研究运用分子生物学技术描述即食虾中两种致病菌混合培养的菌量变化,探究混菌模式下微生物的生长失活情况,为分子预测模型的进一步研究提供了新的思路。     

Antimicrobial activity of Ginkgo biloba leaf extract on Listeria monocytogenes

ABSTRACT: The antimicrobial activities of Ginkgo biloba leaf extract (GBE) and the combined effects of GBE and sodium EDTA (sodium Ethylenediaminetetraacetic acid) against Listeria monocytogenes were determined at 4 °C, 25 °C, and 37 °C. Listeria monocytogenes grown at 37 °C for 24 h was inoculated (6 to 7 log CFU/mL) into BHI broth containing either GBE or GBE and EDTA (1.6 mg/mL) with various GBE concentrations of 0.1, 0.25, 0.5, 1, 2.5, 5.0, 7.5, 10.0, 15.0, or 20.0% vol/vol and stored at 4 °C, 25 °C, and 37 °C. The inhibitory effect of the GBE was more pronounced at low temperature of 4 °C. GBE was effective in inhibiting microbial growth. Addition of EDTA enhanced antimicrobial activity of GBE.     

Growth of Listeria monocytogenes on vacuum-packaged horsemeat for human consumption

In order to investigate the likelihood of Listeria monocytogenes (serotype 4b, ATCC 19115) growth on vacuum-packaged horsemeat at refrigeration temperature, fourteen horsemeat surface/volume homogeneous 150 g weight pieces were superficially inoculated with serotype 4b L. monocytogenes and vacuum packaged. The samples were stored at 4 ± 1 °C. Two pieces (one for pH determination and one for L. monocytogenes counts) were examined at days 0, 7, 14, 21, 28, 35 and 42. Surface pH did not show significant variations during the experiment. The average L. monocytogenes initial contamination level was 1.77log₁₀ CFU/g. A lag phase of 7 days was recorded. The exponential growth rate between day 7 to day 35 was 0.125log₁₀ CFU/day, corresponding to 3.51log₁₀ CFU/g in 28 days. At the end of the experiment the mean L. monocytogenes log₁₀ CFU/g was 5.78.     

Effect of Salt Reduction on Growth of Listeria monocytogenes in Meat and Poultry Systems

Abstract: Reducing sodium in food could have an effect on food safety. The objective was to determine differences in growth of Listeria monocytogenes in meat and poultry systems with salt substitutes. For phase 1, fresh ground beef, pork, and turkey with NaCl, KCl, CaCl₂, MgCl₂, sea salt, or replacement salt added at 2.0% were inoculated with L. monocytogenes to determine growth/survival during 5 d at 4 °C to simulate a pre‐blend process. L. monocytogenes populations significantly decreased (0.41 log CFU/g) during the storage time in beef, but no differences (P > 0.05) were observed over time in pork or turkey. Salt type did not affect (P > 0.05) L. monocytogenes populations during pre‐blend storage. MgCl₂ and NaCl allowed significant growth of aerobic populations during storage. For phase 2, emulsified beef and pork products were processed with 2% NaCl, KCl, sea salt, or a NaCl/KCl blend and post‐process surface‐inoculated with L. monocytogenes to determine growth/survival at 4 °C for 28 d. Pork products showed significantly greater L. monocytogenes population growth at all sampling times (0, 7, 14, 21, and 28 d) than beef products, but salt type had no effect on L. monocytogenes populations with sampling times pooled for data analysis. Although salt types had no impact on L. monocytogenes populations in preblend and emulsified meat products, pork and turkey preblends and emulsified pork had greater L. monocytogenes populations compared with beef products. These studies demonstrate that sodium may not affect the safety of preblends and emulsified meat and poultry products. Practical Application: odium reduction in food is an important topic because of sodium's unfavorable health effects. This research shows that reducing sodium in pre‐blends and emulsified meat and poultry products would have no effect on Listeria monocytogenes populations, but replacement of NaCl with MgCl₂ may affect growth of aerobic populations.     

甘草多糖对单核细胞增生性李斯特菌体内外生长的影响

目的：探讨甘草多糖对单核细胞增生性李斯特菌（Listeria monocytogenes,Lm）体内外生长的影响。方法：采用平板计数法和荧光显微镜、分光光度法及Bliss法分别分析甘草多糖对Lm生长速率和生物被膜形成的影响及计算细菌对小鼠半数致死量（median lethal dose,LD50）。观察低剂量（1 mg/kg）、中剂量（10 mg/kg）和高剂量（100 mg/kg）甘草多糖对感染100×LD50 Lm小鼠的保护力。小鼠感染0.01×LD50的Lm后1、7、14、21、28、35 d剖杀小鼠,检测脾脏、肝脏和肺脏中Lm分布的消长情况。结果：低质量浓度时（10、20、50 μg/mL）,甘草多糖随着质量浓度增加对Lm标准菌株10403S生长速率和生物被膜形成的促进作用呈上升趋势;但高质量浓度时（100、200 μg/mL）这种促进作用逐渐降低。Lm口服感染小鼠的LD50为2.70×108 CFU。高剂量甘草多糖对于100×LD50的Lm感染可提供50%的保护力。0.01×LD50的Lm感染小鼠,随着甘草多糖质量浓度增加小鼠脾脏、肝脏和肺脏的细菌量逐渐减少。结论：低质量浓度甘草多糖在体外对Lm的生长具有促进作用,但随着质量浓度增高促进强度逐渐降低,而甘草多糖可以提高小鼠抵抗Lm感染的能力。     

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.     

Evaluation of Changes in Listeria monocytogenes Populations on Frankfurters at Different Stages from Manufacturing to Consumption

ABSTRACT: This study evaluated the fate of inoculated Listeria monocytogenes on frankfurters stored under conditions simulating those that may be encountered between manufacturing and consumption. Frankfurters with or without 1.5% potassium lactate and 0.1% sodium diacetate (PL/SD) were inoculated (1.8 ± 0.1 log CFU/cm²) with a 10‐strain composite of L. monocytogenes, vacuum‐packaged, and stored under conditions simulating predistribution storage (24 h, 4 °C), temperature abuse during transportation (7 h, 7 °C followed by 7 h, 12 °C), and storage before purchase (60 d, 4 °C; SBP). At 0, 20, 40, and 60 d of SBP, samples were exposed to conditions simulating delivery from stores to homes or food establishments (3 h, 23 °C), and then opened or held vacuum‐packaged at 4 or 7 °C for 14 d (SHF). Pathogen counts remained relatively constant on frankfurters with PL/SD regardless of product age and storage conditions; however, they increased on product without antimicrobials. In vacuum‐packaged samples, during SHF at 4 °C, the pathogen grew faster (P < 0.05) on older product (20 d of SBP) compared to product that was fresh (0 d of SBP); a similar trend was observed in opened packages. At 7 °C, the fastest growth (0.35 ± 0.02 log CFU/cm²/d) was observed on fresh product in opened packages; in vacuum‐packages, growth rates on fresh and aged products were similar. By day 40 of SBP the pathogen reached high numbers and increased slowly or remained unchanged during SHF. This information may be valuable in L. monocytogenes risk assessments and in development of guidelines for storage of frankfurters between package opening and product consumption.     

Inactivation of Listeria monocytogenes on Beef Bologna and Cheddar Cheese Using Polyvinyl-idene Chloride Films Containing Sorbic Acid

ABSTRACT: Polyvinylidene chloride (PVDC, Saran^R F-310) films containing sorbic acid (0%, 1.5%, and 3.0% w/v) were prepared with use of a solvent-casting method and were then placed between slices of commercially produced beef bologna that were previously surface-inoculated with L. monocytogenes at 10³ or 10⁵ CFU/g. In addition, cubes of commercial Cheddar cheese were surface-inoculated to contain 10³ or 10⁵ Listeria monocytogenes colony-forming units (CFU) /g and then wrapped with the sorbic acid-containing films. Films containing 1.5% and 3.0% (w/ v) sorbic acid prevented growth of L. monocytogenes on bologna slices with populations as much as 7.1 logs lower after 28 d of storage at 4 °C compared with the sorbic acid-free controls. In contrast, numbers of Listeria remained relatively stable on Cheddar cheese with populations decreasing < 1.3 logs after 35 d of storage. With use of the sorbic acid-containing films, common spoilage organisms were also inhibited on both products. After 28 d of contact with bologna and Cheddar cheese, these films retained 7% and 60% of their original sorbic acid content, respectively, with the control film retaining 85% of its original sorbic acid content. Given these findings, sorbic acid-containing films may be useful in enhancing the safety and shelf-life of ready-to-eat delicatessen products.     

D and z Values of Salmonella, Listeria innocua, and Listeria monocytogenes in Fully Cooked Poultry Products

: The D and z values of Salmonella, Listeria innocua, and Listeria monocytogenes were obtained for different ready‐to‐eat poultry products, including chicken, turkey, and duck. The D values of Salmonella, L. innocua, and L. monocytogenes were 151.5 to 0.1 min at 55 to 70°C, and the z values of Salmonella, L. innocua, and L. monocytogenes were 4.9 to 7.0 °C. Significant differences were found for the heat resistance of Salmonella, L. innocua, and L. monocytogenes among turkey, duck, and chicken products, indicating that the kinetic values of a certain pathogen in a specific product should be used for determining process lethality in fully cooked and vacuum‐packaged poultry products during post‐cook heat treatments.     

猪肉中单核细胞增生李斯特菌的分离与耐药性研究

了解猪肉中单核细胞增生李斯特菌的污染情况、血清型和耐药性现状。按照国标GB 4789.30-2010程序分离猪肉中单核细胞增生李斯特菌;多重PCR确定血清型;纸片扩散法检测单核细胞增生李斯特菌对抗生素的敏感性。研究结果表明273份猪肉样本中有52（19.02%)个受单核细胞增生李斯特菌的污染,共检出78株该致病菌。33株（42.31%）、24株（30.71%）、16株（20.51%）、3株（3.85%）、2株（2.56%)分别被确定为血清型1/2a（或3a）、1/2b（或3b或7）、1/2c（或3c）、4a（或4c）和4b（或4d或4e）。所有菌株对氨苄西林、 氨苄西林/舒巴坦、亚胺培能、环丙沙星、左氧氟沙星、万古霉素都敏感,对四环素（20.51%）的耐药率最为严重。11.54%的菌株对临床治疗李斯特菌病的青霉素、庆大霉素、红霉素等抗生素出现耐药。tet(M)基因是单核细胞增生李斯特菌耐四环素的主要机制之一,接合型转座子Tn916与该菌四环素耐药散播有直接关系。     

熟肉制品中单核细胞增生李斯特氏菌耐药及致病的遗传分析

目的 获得中国不同省份熟肉制品中的33株单核细胞增生李斯特氏菌(单增李斯特菌)的抗生素敏感性特征图谱,并运用全基因组测序对菌株进行耐药和致病的基因遗传分析.方法 采用微量肉汤稀释法对33株熟肉制品中的单增李斯特菌进行药敏测定,同时进行高精度框架图测序,基因组序列经组装后通过相应的生物信息学流程进行数据分析.结果 33株...     

Survival of Listeria monocytogenes, Listeria innocua, and Lactic Acid Bacteria in Chill Brines

ABSTRACT:  Listeria monocytogenes is the pathogen of concern in ready-to-eat (RTE) meat products. Salt brines are used to chill processed meats. L. monocytogenes and lactic acid bacteria (LAB) can grow under saline conditions, and may compete with each other for nutrients. The objective of this study was to determine the effect of lactic acid bacteria ( Enterococcus faecalis , Carnobacterium gallinarum , and Lactobacillus plantarum ) on the survival of L. monocytogenes and Listeria innocua in brines stored under low temperatures for 10 d. Sterile tap water (STW) and 2 brine solutions (7.9% and 13.2% NaCl) were inoculated with 1 of 5 cocktails ( L. monocytogenes , L. innocua , LAB, L. monocytogenes + LAB, or L. innocua + LAB) at initial concentrations of 7 log CFU/mL. Brines were stored for 10 d at 4 or 12 °C. Three replications of each brine concentration/cocktail/temperature combination were completed. No significant reductions of L. monocytogenes occurred in 7.9%[w/v] or 13.2%[w/v] brines when LAB were present; however, there were significant reductions after 10 d of L. monocytogenes in the STW solution when LAB were present (1.43 log CFU/mL at 4 °C and 3.02 log CFU/mL at 12 °C). L. innocua was significantly less resilient to environmental stresses of the brines than L. monocytogenes , both with and without LAB present ( P ≤ 0.05). These strains of lactic acid bacteria are not effective at reducing L. monocytogenes in brines at low temperatures. Furthermore, use of L. innocua as a model for L. monocytogenes is not appropriate under these environmental conditions.     

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。      

携带质粒李斯特菌耐药及重金属抵制特性研究

鉴定单核细胞增生性李斯特菌(Lm)携带质粒特性,分析携带质粒Lm对抗生素、重金属镉及苯扎氯铵的敏感性。结果表明,11株Lm携带内源性质粒;所有Lm菌株对阿莫西林、红霉素和利福平均敏感(22/22),对头孢拉定100%耐药(22/22);其中5株对盐酸万古霉中度耐受,部分菌株对盐酸环丙沙星(17/22)、硫酸新霉素(16/22)及盐酸四环素素(9/22)具有一定耐受性。耐镉分析表明,菌株对重金属镉呈现抵制特性(14/22),且均为携带质粒菌株;而在苯扎氯铵敏感性检测中,仅3株携带质粒Lm菌株对苯扎氯铵具有抵制性。对Lm菌株进行噬菌体杀菌检测,结果均能够被噬菌体识别并裂解。综上所述,Lm分离株耐药性增强,且对苯扎氯铵和镉有抵制特性,而噬菌体能够杀灭抵制菌株,为控制耐药及抵制性菌株污染提供了新途径。     

Detection, Enumeration, and RAPD Analysis of Listeria monocytogenes Isolates in Fish Derived fromRetail Outlets in WesternMassachusetts

Fish were sampled over a 24-month period from two major supermarket retail outlets in Hadley, Massachusetts, USA, designated A and B, for the incidence of Listeria monocytogenes and numbers of the organism present per 100 g of tissue. Fifteen species of fish were represented. Seventy-four samples out of a total of 320 were confirmed by PCR as yielding L. monocytogenes. From retail source A, a total of 171 samples yielded 59 (34.5%) that were positive for the presence of L. monocytogenes. In contrast, from retail source B, a total of 149 samples yielded 15 (10.0%) that were positive. Only six samples (3.5%) from retail source A had MPN counts of L. monocytogenes in the range of 100 to 1,000 per 1,000 g. Only two samples (1.3%) from retail source B had counts of L. monocytogenes from 100 to 1,000 per 100 g. A total of 221 strains of L. monocytogenes were derived from the MPN cultures, 164 from retail source A, and 57 from retail source B. All 221 strains were subjected to RAPD analysis using three random primers. Primer LMPB1 yielded 21 RAPD profiles, primer LMPB4 yielded 19 profiles, and primer HLWL74 yielded 26 profiles. A total of 55 composite profiles were identified by combining the profiles derived from the three primers. Source A yielded 50 composite RAPD profiles, whereas source B yielded only ten composite profiles. In addition, 27 of the 55 composite profiles were derived from individual isolates and RAPD types 11 and 18 included 49 and 27 isolates respectively. Fish from retail source A clearly harbored far more RAPD types than did source B. The results clearly indicated that two major retail sources in close geographic proximity can vary considerably with respect to the incidence and numbers of L. monocytogenes present on the fish tissue. It was not possible to determine whether the processors furnishing fish to retail outlet A or the supermarket itself was responsible for the notably higher incidence and numbers of L. monocytogenes on fish from retail source A compared to fish from retail source B.     

抗菌肽zp37抑制果汁中单核细胞增生李斯特菌的活性及其作用机制

单核细胞增生李斯特菌（简称单增李斯特菌）作为一种常见的食源性致病菌,是影响果汁食用安全性的主要威胁因子。本实验设计并获得一种抗菌肽zp37,通过测定抗菌肽zp37最小抑菌浓度（minimum inhibitory concentration,MIC）,分析抗菌肽zp37处理后单增李斯特菌的生长曲线、膜电位、膜完整性、细胞微观结构、细胞聚集率、DNA结合情况以及胞内活性氧等,研究抗菌肽zp37抑制果汁中单增李斯特菌的活性及其作用机制。结果表明,抗菌肽zp37对单增李斯特菌的MIC为16μg/mL,当其质量浓度大于4MIC时,单增李斯特菌生长受到明显抑制。同时,8MIC的抗菌肽zp37处理1h能杀死绝大部分的单增李斯特菌,且在7 d贮藏期内,2MIC的抗菌肽zp37能将草莓汁、猕猴桃汁和苹果汁中的单增李斯特菌控制到检测限以下。抗菌肽zp37处理后,单增李斯特菌细胞膜离子通透性增加、膜电位消失,细胞膜完整性被破坏,细胞出现明显的凹陷变形,细胞间发生聚集。荧光标记示踪法结果显示,抗菌肽zp37同时作用于单增李斯特菌的细胞膜和细胞质。在细胞质中,当抗菌肽zp37质量浓度大于DNA质量浓度的1/4...