绿豆芽生产过程中微生物的生长、分布及消毒剂处理效果评价

研究绿豆芽生产过程中微生物的生长情况及种类分布,采用2种不同类型的消毒剂进行处理及效果评价,并用16SrRNA方法对豆芽中主要微生物进行鉴定。结果表明,成品绿豆芽中的微生物总量为5.8×10~7 CFU/g,微生物的种类主要为克雷伯氏菌(Klebsiella)、不动杆菌(Acinetobacter)、肠杆菌(Enterobacter)和假单胞菌属(Pseudomonas)等。有效氯浓度1 000~3 000 mg/L的NaClO处理芽菜30 min,菌落总数分别降至1.2×10~7~1.0×10~5 CFU/g,而直接处理单独培养的模型对照组则平均降至2.3×10~4~1.0×10~3 CFU/mL。25 mg/L AgNO_3可使菌落总数降至3.8×10~5 CFU/g,消毒效果同样差于模型对照组。扫描电子显微镜观察分析芽菜表面微生物可形成明显可见的生物膜,显著强化了细菌对消毒剂的耐受力。     

柠檬草精油纳米脂质体抗不同材质表面金黄色葡萄球菌生物膜的研究

主要研究了柠檬草精油及其纳米脂质体对不同材质表面金黄色葡萄球菌生物膜的抑制和清除作用。在柠檬草精油对金黄色葡萄球菌生物膜的抑制和清除实验中,抑制结果表明,当柠檬草精油浓度达到2 mg/m L时,不锈钢片表面无生物膜生成,精油浓度达到4 mg/m L时,塑料和玻璃片表面的生物膜被完全抑制;清除结果表明,8 h时,4 mg/m L的精油对不锈钢、塑料片和玻璃片表面的生物膜清除率分别达到了99.91%、99.63%和99.94%。通过薄膜超声波分散法制备了柠檬草精油纳米脂质体,脂质体平均粒径为168.5nm,PDI为0.276,Zeta电位为-50.62 m V,包封率为31.49%。精油脂质体对不同材质表面的金黄色葡萄球菌生物膜均具有良好的抗生物膜性能,当作用15天时,不锈钢、塑料和玻璃片表面生物膜的清除率分别达到99.89%、99.99%和99.81%。     

肉桂精油对蔬菜表面大肠杆菌O157∶H7生物膜的杀菌作用研究

实验评价了肉桂精油对大肠杆菌O157:H7的杀菌活性,并且研究了肉桂精油对新鲜蔬菜表面大肠杆菌O157:H7生物膜的清除效果。实验结果表明,肉桂精油具有较强的杀菌活性,短时间内即可灭活高浓度的大肠杆菌O157:H7游离细菌。此外,肉桂精油对大肠杆菌O157:H7生物膜有较好的清除作用,其最小抑制生物膜浓度(MBIC)和最小清除生物膜浓度(MBEC)仅为0.5 mg/m L和1 mg/m L,在浓度为2 mg/m L及4 mg/m L时,可在60 min内完全灭活生物膜细菌。当肉桂精油应用于生菜、菠菜、莴苣、青椒及黄瓜表面时,大肠杆菌O157:H7生物膜细菌数量大幅降低。附着在蔬菜表面的生物膜经4 mg/m L精油处理2d后,细菌数量均低于10 CFU/cm2。     

5种氧化型减菌剂对罗非鱼片品质及蛋白质的影响

本研究以罗非鱼片为原料,考察5种常见氧化型减菌剂(H₂O₂溶液、ClO₂溶液、NaClO溶液、臭氧水和微酸性电解水)的减菌效果及其对鱼片质构特性、色泽及肌原纤维蛋白(myofibrillar protein,MP)氧化情况的影响。结果表明,减菌率达80%时各氧化型减菌剂的质量浓度和作用时间分别为：1 500 mg/L H₂O₂溶液浸泡8 min;200 mg/L ClO₂溶液浸泡10 min;200 mg/L NaClO溶液浸泡10 min;9 mg/L臭氧水浸泡10 min;30 mg/L微酸性电解水浸泡20 min。在上述5种减菌条件下对鱼肉色泽、质构特性、MP氧化情况与对照组进行比较,发现减菌处理后的罗非鱼片硬度、L*值增大,a*值减小,MP质量浓度、总巯基含量下降,羰基含量升高,表面疏水性基团数量增加,其中微酸性电解水和NaClO溶液处理组蛋白质的变性程度最明显;蛋白荧光强度降低,其中微酸性电解水处理组降低幅度最大;MP的α-螺旋和β-转角相对含量...     

丁香酚对解淀粉芽孢杆菌气液界面生物膜的抑制作用

为研究丁香酚对解淀粉芽孢杆菌DY1a生物膜的抑制作用及成膜早期界面黏附聚集能力的影响,分析了不同质量浓度的丁香酚对生物膜形成、生物膜表面微观结构、膜内活菌数及生物膜基质多糖和蛋白质含量的影响,并评价其对成熟生物膜的清除能力,通过细菌运动能力实验、细胞表面疏水性、细胞表面Zeta电位及细胞自聚集能力,综合分析丁香酚对生物膜形成早期界面黏附聚集能力的影响。结果表明:丁香酚对解淀粉芽孢杆菌的最低生物膜抑制质量浓度(MBIC)为1.500mg/mL,MBIC的丁香酚对成熟生物膜清除率为28.85%。添加1/2 MBIC、1/4 MBIC的丁香酚培养基气液界面形成的生物膜较为单薄,表面较为光滑平整,生物膜内活菌数显著降低。丁香酚对腐败菌泳动能力抑制率为22.16%~100.00%,对丛集能力的抑制率为43.86%~97.50%,使细胞表面疏水性、细胞表面负Zeta电位和自聚集率均降低。此外,丁香酚显著抑制了腐败菌胞外多糖和蛋白质合成。丁香酚对芽孢杆菌生物膜的抑制作用主要是通过抑制细菌运动能力,降低细胞表面的疏水性、自聚集性,从而干扰早期菌体在成膜界面上的黏附能力,并通过抑制胞外聚合物组分的合成分泌延迟生物膜的形成和成熟。丁香酚可作为抑制腐败解淀粉芽孢杆菌气液界面生物膜形成的潜在抗生物膜剂。     

嗜热链球菌和保加利亚乳杆菌混菌生物膜发酵特性及其抗逆性

利用扫描电子显微镜对嗜热链球菌(Streptococcus thermophilus)和保加利亚乳杆菌(Lactobacillus bulgaricus)在不锈钢网布及椰果粒表面形成的混菌生物膜进行观察,并对不锈钢网布表面混菌生物膜发酵特性及其抗逆性进行研究。结果表明:Streptococcus thermophilus和Lactobacillus bulgaricus可在两种载体表面形成典型的混菌生物膜结构,且不锈钢网布表面混菌生物膜的发酵液菌体密度变化曲线与细菌S型生长曲线相似,但在停滞期和对数期不明显,发酵液pH值呈现先下降后趋于平稳的趋势,终止pH值接近4.0;此外,不锈钢网布表面混菌生物膜比游离乳酸菌具有更强的抗性,并且可在15d内稳定地释放较高浓度的游离乳酸菌。     

保加利亚乳杆菌和嗜热链球菌生物膜形成研究

采用置片法和菌落计数法对嗜热链球菌(Streptococcus thermophilus)和保加利亚乳杆菌(Lactobacillus bulgaricus)在不同载体表面(椰果粒、不锈钢网布、塑料片、陶瓷片和玻璃片)上形成生物膜的能力进行研究。结果表明：椰果粒和不锈钢网布是适宜乳酸菌单菌生物膜形成的载体,培养7d椰果粒和不锈钢网布上的单菌生物膜初始菌密度可达107CFU/cm2,并且在后续培养中能稳定在3.2×106CFU/cm2左右;混菌生物膜菌密度连续7d稳定在1×107CFU/cm2左右。利用扫描电镜对S. thermophilus和L. bulgaricus形成的混菌生物膜进行观察,结果表明在椰果粒和不锈钢网布表面上S. thermophilus和L. bulgaricus形成典型的混菌生物膜结构。     

不同培养基质上阪崎克罗诺杆菌生物膜的比较

为了比较阪崎克罗诺杆菌在玻璃、聚丙烯及不锈钢表面形成的生物膜,将培养48 h的阪崎克罗诺杆菌生物膜经荧光染料SYTO 9/PI标记后,用激光共聚焦扫描显微镜(confocal laser scanning microscope,CLSM)联合COMSTAT软件定性定量地分析生物膜结构。结果表明:阪崎克罗诺杆菌在3种材料表面形成的生物膜的总生物量、平均厚度及平均扩散距离依次是不锈钢聚丙烯玻璃,粗糙系数及表面积与生物量比值则为不锈钢聚丙烯玻璃,生物膜的菌体活死比例是玻璃高于聚丙烯、不锈钢。另外,标准菌株ATCC 29544生物膜的总生物量、菌体活死比例、平均厚度及平均扩散距离均高于分离株Ⅰ、Ⅱ,粗糙系数及表面积与生物量比值均低于分离株。可见,阪崎克罗诺杆菌在不锈钢表面的成膜性强于玻璃与聚丙烯,标准菌株ATCC 29544的成膜能力高于2株食品分离菌株。     

阪崎肠杆菌生物膜形成条件及超声波对其去除作用

通过96/6孔板培养法进行生物膜体外模型建立,比浊法测定生物膜含量,结晶紫染色法观察生物膜形态,平板菌落计数法评价超声波对玻璃表面生物膜的去除作用效果,激光共聚焦显微镜观察超声波处理后生物膜脱落效果,研究阪崎肠杆菌生物膜的形成条件和超声波处理对其去除作用效果。结果表明,阪崎肠杆菌生物膜形成的最适温度37℃,最适pH 7.5,最适盐度为2.5%。CaCl_2(0.50%)、MgCl_2(1.50%)对阪崎肠杆菌生物膜形成具有明显促进作用。Cu~(2+)(1.50%)、EDTA(2.5%)对阪崎肠杆菌生物膜形成具有抑制作用。超声波处理能够使玻璃表面上附着的阪崎肠杆菌生物膜脱落,超声波处理初始温度30℃,时间14 min条件下去除效果最好。激光共聚焦显微镜可直接观察到超声波处理后盖玻片上生物膜中阪崎肠杆菌活菌数量变化,可用于证实和评价超声处理对玻璃表面阪崎肠杆菌生物膜的去除效果。     

单增李斯特菌生物膜形成及其调控机制研究进展

单增李斯特菌是一种重要的食源性致病菌,极易在食品接触表面形成难以清除的生物膜,导致食品持续性的污染。细菌生物膜在形成量、活细菌数量、微观结构等方面受到环境因素及菌株自身特性的影响。在单增李斯特菌的生物膜形成过程中,多种调控机制发挥着重要的作用。该文介绍了细菌生物膜的形成过程及影响单增李斯特菌生物膜形成的重要因素,简述了与单增李斯特菌生物膜相关的基因调控、胞外聚合物质分泌与群体感应系统调节的研究进展,有助于更好地理解其生物膜形成的复杂生理过程,为单增李斯特菌生物膜的污染及防控提供一定参考。     

Assessment of a regulatory sanitization process in Egyptian dairy plants in regard to the adherence of some food-borne pathogens and their biofilms

Food-borne pathogens may develop certain strategies that enable them to defy harsh conditions such as chemical sanitization. Biofilm formation represents a prominent one among those adopted strategies, by which food-borne pathogens protect themselves against external threats. Thus, bacterial biofilm is considered as a major hazard for safe food production. This study was designed to investigate the adherence and the biofilm formation ability of some food-borne pathogens on stainless steel and polypropylene surfaces using chip assay, and to validate regular sanitizing process (sodium hypochlorite 250mg/L) for effective elimination of those pathogens. Sixteen pathogenic bacterial strains, previously isolated from raw milk and dairy products at Zagazig city, Egypt (9 Staphylococcus aureus, 4 Cronobacter sakazakii and 3 Salmonella enterica serovar Typhimurium), were chosen for this study. Strains showed different patterns of adherence and biofilm formation on tested surfaces with minor significance between surfaces. The ability of sodium hypochlorite to completely eradicate either adhered or biofilm-embedded pathogens varied significantly depending on the strain and type of surface used. Whilst, sodium hypochlorite reduced tested pathogens counts per cm(2) of produced biofilms, but it was not able to entirely eliminate neither them nor adherent Cronobacter sakazakii to stainless steel surface. This study revealed that biofilm is considered as a sustainable source of contamination of dairy products with these pathogens, and also emphasized the need of paying more attention to the cleaning and sanitizing processes of food contact surfaces.     

四种不同接触表面蜡样芽孢杆菌菌膜形成的影响

为了解食源性致病菌蜡样芽孢杆菌在食品加工环境中菌膜形成能力,以玻璃、不锈钢、聚氯乙烯、聚丙烯为接触面,采用超声波平板菌落计数法测定不同环境因素(温度、pH、氯化钠、葡萄糖、苯甲酸钠及山梨酸钾)、不同材料表面蜡样芽孢杆菌(B.cereus)菌膜形成的变化趋势。结果表明:四种材质表面形成B.cereus菌膜能力的大小顺序为:玻璃 > 不锈钢 > 聚氯乙烯 > 聚丙烯。其中,30 ℃,pH7.0时菌膜形成量最大,添加低浓度葡萄糖(4.0%)或氯化钠(0.5%)对B.cereus菌膜形成有显著促进作用(p<0.05),添加0.15%苯甲酸钠、山梨酸钾的菌膜形成量显著高于添加0.10%的菌膜形成量(p<0.05)。本研究为蜡样芽孢杆菌风险评估提供基础数据,为食品工业蜡样芽孢杆菌菌膜的预防和控制奠定基础,为改进蜡样芽孢杆菌的清洗控制措施提供参考。     

ε- 聚赖氨酸在冷鲜猪肉保鲜中的应用

研究ε- 聚赖氨酸(ε-PL)保鲜冷鲜猪肉的处理条件,考察在优化条件下ε-PL 对冷鲜猪肉感官、微生物、理化等指标的影响。结果表明：400mg/L ε-PL 单独使用时就能显著抑制冷鲜猪肉感官品质的下降、微生物的生长繁殖、pH 值的上升和TVB-N 的积累,而当ε-PL 和乙酸复配使用后抑制作用更佳,特别是对微生物的生长繁殖有明显的抑制作用,浸泡时间对ε-PL- 乙酸助剂的抑菌效果影响很大,随着浸泡时间的增加,ε-PL- 乙酸助剂抑菌效果逐渐加强。综合抑菌效果、处理成本以及处理效率考虑,优化组合为：ε-PL 质量浓度400mg/L,助剂为0.2% 乙酸,浸泡时间为5min。优化处理后冷鲜猪肉保质期从对照组2d 延长至8d。结果揭示：ε-PL 作为冷鲜肉生物保鲜剂,具有广阔的应用前景。     

Efficacy of neutral electrolyzed water to inactivate Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, and Staphylococcus aureus on plastic and wooden kitchen cutting boards

This study evaluated the efficacy of neutral electrolyzed water (NEW; 64.1 mg/liter of active chlorine) to reduce populations of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Listeria monocytogenes on plastic and wooden kitchen cutting boards. Its effectiveness was compared with that of a sodium hypochlorite solution (NaClO; 62.3 mg/liter of active chlorine). Inoculated portions of cutting boards were rinsed in either NEW or NaClO solutions, or deionized water (control). Plastic boards were rinsed for 1 min and wooden boards for 1 and 5 min. After each treatment, the surviving population of each strain was determined on the surface and in the soaking water. No significant difference (P > or = 0.05) was found between the final populations of each strain with regard to the treatment solutions (NEW or NaClO). However, a significant difference (P < or = 0.05) was revealed between surface materials after 1 min of washing. Whereas in plastic boards the initial bacterial populations were reduced by 5 log CFU/50 cm2, in wooden cutting boards they underwent a reduction of <3 log CFU/50 cm2. A 5-min exposure time yielded reductions of about 4 log CFU/50 cm2. The surviving populations of all bacteria in NEW and NaCIO washing solutions were <1 log CFU/ml after soaking both surfaces. This study revealed that NEW treatment is an effective method for reducing microbial contamination on plastic and wooden cutting boards. NEW efficacy was comparable to that of NaCIO, with the advantage of having a larger storage time.     

Biofilm formation by multidrug-resistant Salmonella enterica serotype typhimurium phage type DT104 and other pathogens

The biofilm-forming capability of Salmonella enterica serotypes Typhimurium and Heidelberg, Pseudomonas aeruginosa, Listeria monocytogenes, Escherichia coli O157:H7, Klebsiella pneumoniae, and Acinetobacter baumannii isolated from humans, animal farms, and retail meat products was evaluated by using a microplate assay. The tested bacterial species showed interstrain variation in their capabilities to form biofilms. Strong biofilm-forming strains of S. enterica serotypes, E. coli O157: H7, P. aeruginosa, K. pneumoniae, and A. baumannii were resistant to at least four of the tested antibiotics. To understand their potential in forming biofilms in food-processing environments, the strong biofilm formers grown in beef, turkey, and lettuce broths were further investigated on stainless steel and glass surfaces. Among the tested strains, Salmonella Typhimurium phage type DT104 (Salmonella Typhimurium DT104) isolated from retail beef formed the strongest biofilm on stainless steel and glass in beef and turkey broths. K. pneumoniae, L. monocytogenes, and P. aeruginosa were also able to form strong biofilms on the tested surface materials. Salmonella Typhimurium DT104 developed a biofilm on stainless steel in beef and turkey broths through (i) initial attachment to the surface, (ii) formation of microcolonies, and (iii) biofilm maturation. These findings indicated that Salmonella Typhimurium DT104 alongwith other bacterial pathogens could be a source of cross-contamination during handling and processing of food.     

蒲公英总黄酮提取物在冷鲜猪肉涂膜保鲜中的应用

选用蒲公英总黄酮提取物和壳聚糖为涂膜材料,研究蒲公英总黄酮提取物对冷鲜肉的保鲜作用。考察蒲公英总黄酮提取物对冷鲜肉感官、理化、微生物等指标的影响。结果表明：蒲公英总黄酮提取物和壳聚糖分别为0.45 mg/L与2.0 g/100 mL时,浸泡时间5 min,冷鲜肉的挥发性盐基总氮含量、pH值、H2S实验和菌落总数的变化能得到有效控制。优化处理后冷鲜肉的保鲜期与对照组比较可延长8 d。证明由壳聚糖和蒲公英总黄酮提取物构成的复合天然保鲜剂具有抑制微生物生长、延长冷鲜肉货架期的作用。     

A predictive model for heat inactivation of Listeria monocytogenes biofilm on stainless steel

Heat treatment of potential biofilm-forming sites is sometimes used for control of Listeria monocytogenes in food processing plants. However, little information is available on the heat treatment required to kill L. monocytogenes present in biofilms. The purpose of this study was to develop a predictive model for the heat inactivation of L. monocytogenes in monoculture biofilms (strains Scott A and 3990) and in biofilms with competing bacteria (Pseudomonas sp. and Pantoea agglomerans) formed on stainless steel in the presence of food-derived soil. Biofilms were produced on stainless steel coupons with diluted tryptic soy broth incubated for 48 h at 25 degrees C. Duplicate biofilm samples were heat treated for 1, 3, 5, and 15 min at 70, 72, 75, 77, and 80 degrees C and tested for survivors using enrichment culture. The experiment was repeated six times. A predictive model was developed using logistic regression analysis of the fraction negative data. Plots showing the probability of L. monocytogenes inactivation in biofilms after heat treatment were generated from the predictive equation. The predictive model revealed that hot water sanitation of stainless steel can be effective for inactivating L. monocytogenes in a biofilm on stainless steel if time and temperature are controlled. For example, to obtain a 75% probability of total inactivation of L. monocytogenes 3990 biofilm, a heat treatment of 80 degrees C for 11.7 min is required. The model provides processors with a risk management tool that provides predicted probabilities of L. monocytogenes inactivation and allows a choice of three heat resistance assumptions. The predictive model was validated using a five-strain cocktail of L. monocytogenes in the presence of food soil.     

冷却猪肉及托盘表面细菌生物被膜分析和肉源荧光假单胞菌的鉴定与被膜研究

为研究冷却猪肉及其接触面中细菌生物被膜能力并探讨肉类特定腐败菌的生物被膜特征,分析了腐败的冷却肉和销售托盘表面的细菌生物被膜形成能力;利用形态学观察、16S r DNA分析和VITEK2微生物鉴定系统鉴定了冷却肉中一株生物被膜能力较强的荧光假单胞菌(Pseudomonas fluorescens);测定了该菌在培养基和猪肉浸提液中的生物被膜能力;用激光共聚焦和扫描电子显微镜观测了其生物被膜的结构特征。结果发现:冷却肉中及其销售托盘表面的细菌能够形成生物被膜的比例较高,37%和45%的细菌具有较强生物被膜形成能力。肉源性荧光假单胞菌能够在培养基和肉液中形成大量生物被膜,具有较强的生物被膜能力。显微成像结果表明荧光假单胞菌在培养6 h后有明显黏附,18 h后多糖分泌增多、菌体堆积并形成具有生物功能的立体生物被膜。这些特点可能有利于荧光假单胞菌在冷却肉表面黏附并成为优势腐败菌。     

绿色魏斯氏菌发酵液对单核细胞增生李斯特菌的抑制及其在冷却猪肉保鲜中的应用

采用微量肉汤稀释法测定绿色魏斯氏菌(Weissella viridescens)发酵液对单核细胞增生李斯特菌(Listeria monocytogenes,Lm)的最小抑菌浓度,微孔板法结合显微镜观察检测绿色魏斯氏菌发酵液对Lm生物被膜形成的影响,最后通过测定冷藏过程中冷却猪肉的菌落总数、Lm总数、感官指标和理化指标评...