Inhibition of foodborne bacteria by the lactoperoxidase system in a beef cube system

Biopreservatives are being developed to inhibit the growth of foodborne pathogens and thus improve food safety. The lactoperoxidase system (LPS) is a naturally occurring system that has potential for use as an antimicrobial agent in foods. Growth of single strains of the pathogens Staphylococcus aureus, Listeria monocytogenes, Escherichia coli O157:H7, Salmonella enterica subsp. enterica serovar Typhimurium, Yersinia enterocolitica, Pseudomonas aeruginosa and beef microflora were assessed on LPS-treated meat surfaces in an experimental system. Beef cubes inoculated with approximately 10(4) cfu cm(-2) of bacteria were treated with the LPS and incubated at 37 degrees C for 24 h, 12 degrees C for 7 days or in a chilling regime: 12 to -1 degrees C over 1 week and held at -1 degrees C for 4 weeks. Treatment with LPS was more effective at storage temperatures non-permissive for rapid bacterial growth with strong inhibition of growth achieved on LPS-treated cubes at 12 degrees C and reduction in pathogen viable counts at chilling temperatures. At chilling temperatures, the LPS inhibited the growth of native pseudomonads but did not prevent the development of native lactic acid bacteria.     

Inhibition of bacterial foodborne pathogens by the lactoperoxidase system in combination with monolaurin

The lactoperoxidase system (LPS) and monolaurin (ML) are potential natural antimicrobial agents for use in foods. The LPS is considered to have greatest activity against Gram-negative bacteria while ML is usually considered to have greatest activity against Gram-positive bacteria. An LPS-ML combination system (utilizing lactoperoxidase (LPX) in the range 5-200 mg kg(-1) and ML in the range 50-1,000 ppm) inhibited growth of Escherichia coli O157:H7 and Staphylococcus aureus. Growth of S. aureus was inhibited more strongly in broth than in milk, in milk than in ground beef A similar pattern was observed for E. coli O157:H7, though enhanced inhibition by LPS-ML systems over that obtained in comparable LPS only systems was not observed in ground beef The inhibitory action of the LPS in combination with other lipids was also examined, with progressively weaker inhibition observed in combinations including palmitoleic acid, monopalmitolein, lauric acid, caprylic acid, and sodium lauryl sulphate.     

Antibacterial effect of the lactoperoxidase system on psychotrophic bacteria in milk.

Activation of the antibacterial lactoperoxidase system in milk, i.e. increasing the thiocyanate concentration to 0.25 mM and adding an equimolar amount of H2O2, results in a substantial reduction of the bacterial flora and prevents the multiplication of psychrotrophic bacteria for up to 5 d. This treatment has no effect on the physico-chemical properties of milk and does not lead to the accumulation of resistant bacteria. The practical application of the lactoperoxidase system in prolonging the storage period of raw milk at low temperatures is discussed.     

Effect of high-pressure treatment on denaturation of bovine lactoferrin and lactoperoxidase

Lactoferrin and lactoperoxidase are whey proteins with biological properties that may provide health benefits to consumers. These properties are vulnerable to potentially denaturing conditions during processing. High-pressure treatment is an appealing alternative to the traditional heat processing of foods because it exerts an antimicrobial effect without changing the sensory and nutritional quality of foods. In this work, the effect of high-pressure treatment on the denaturation of lactoferrin and lactoperoxidase present in skim milk and whey, and as isolated proteins in buffer, was studied over a pressure range of 450 to 700 MPa at 20°C. Denaturation of lactoferrin was measured by the loss of reactivity with their specific antibodies using a sandwich ELISA. Denaturation of lactoperoxidase was determined by measuring the loss of enzymatic activity using a spectrophotometric technique. No substantial inactivation of lactoperoxidase was observed in any treatment assayed. The concentration of the residual immunoreactive lactoferrin after each pressure treatment was determined, and the data were subjected to kinetic analysis to obtain D and Z values. Denaturation of lactoferrin increased with pressure and holding time, and D values were lower when lactoferrin was treated in whey than in milk, and lower in both whey and milk than in phosphate buffer. Thus, protein is denatured more slowly in buffer and in milk than in whey. Denaturation of lactoferrin in the 3 media was found to follow a reaction order of n=1.5. Volumes of activation of about -34.77, -24.35, and -24.09 mL/mol were obtained for lactoferrin treated in skim milk, whey, and buffer, respectively, indicating a decrease in protein volume under pressure.     

Challenge testing the lactoperoxidase system against a range of bacteria using different activation agents

Lactoperoxidase (LP) exerts antimicrobial effects in combination with H₂O₂ and either thiocyanate (SCN⁻) or a halide (e.g., I⁻). Garlic extract in the presence of ethanol has also been used to activate the LP system. This study aimed to determine the effects of 3 LP activation systems (LP+SCN⁻+H₂O₂; LP+I⁻+H₂O₂; LP + garlic extract + ethanol) on the growth and activity of 3 test organisms (Staphylococcus aureus, Pseudomonas aeru-ginosa, and Bacillus cereus). Sterilized milk was used as the reaction medium, and the growth pattern of the organisms and a range of keeping quality (KQ) indicators (pH, titratable acidity, ethanol stability, clot on boiling) were monitored during storage at the respective optimum growth temperature for each organism. The LP+I⁻+ H₂O₂ system reduced bacterial counts below the detection limit shortly after treatment for all 3 organisms, and no bacteria could be detected for the duration of the experiment (35 to 55 h). The KQ data confirmed that the milk remained unspoiled at the end of the experiments. The LP + garlic extract + ethanol system, on the other hand, had no effect on the growth or KQ with P. aeruginosa, but showed a small retardation of growth of the other 2 organisms, accompanied by small increases (5 to 10 h) in KQ. The effects of the LP+SCN⁻+H₂O₂ system were intermediate between those of the other 2 systems and differed between organisms. With P. aeruginosa, the system exerted total inhibition within 10 h of incubation, but the bacteria regained viability after a further 5 h, following a logarithmic growth curve. This was reflected in the KQ indicators, which implied an extension of 15 h. With the other 2 bacterial species, LP+SCN⁻+H₂O₂ exerted an obvious inhibitory effect, giving a lag phase in the growth curve of 5 to 10 h and KQ extension of 10 to 15 h. When used in combination, I⁻ and SCN⁻ displayed negative synergy.     

Radiation inactivation of foodborne pathogens on frozen seafood products

Foodborne illness due to consumption of contaminated seafood is, unfortunately, a regular occurrence in the United States. Ionizing (gamma) radiation can effectively inactivate microorganisms and extend the shelf life of seafood. In this study, the ability of gamma irradiation to inactivate foodborne pathogens surface inoculated onto frozen seafood (scallops, lobster meat, blue crab, swordfish, octopus, and squid) was investigated. The radiation D(10)-values (the radiation dose needed to inactivate 1 log unit of a microorganism) for Listeria monocytogenes, Staphylococcus aureus, and Salmonella inoculated onto seafood samples that were then frozen and irradiated in the frozen state (-20°C) were 0.43 to 0.66, 0.48 to 0.71, and 0.47 to 0.70 kGy, respectively. In contrast, the radiation D(10)-value for the same pathogens suspended on frozen pork were 1.26, 0.98, and 1.18 kGy for L. monocytogenes, S. aureus, and Salmonella, respectively. The radiation dose needed to inactivate these foodborne pathogens on frozen seafood is significantly lower than that for frozen meat or frozen vegetables.     

Innovative nonthermal technologies for inactivation of emerging foodborne viruses

Various foodborne viruses have been associated with human health during the last decade, causing gastroenteritis and a huge economic burden worldwide. Furthermore, the emergence of new variants of infectious viruses is growing continuously. Inactivation of foodborne viruses in the food industry is a formidable task because although viruses cannot grow in foods, they can survive in the food matrix during food processing and storage environments. Conventional inactivation methods pose various drawbacks, necessitating more effective and environmentally friendly techniques for controlling foodborne viruses during food production and processing. Various inactivation approaches for controlling foodborne viruses have been attempted in the food industry. However, some traditionally used techniques, such as disinfectant-based or heat treatment, are not always efficient. Nonthermal techniques are considered a new platform for effective and safe treatment to inactivate foodborne viruses. This review focuses on foodborne viruses commonly associated with human gastroenteritis, including newly emerged viruses, such as sapovirus and Aichi virus. It also investigates the use of chemical and nonthermal physical treatments as effective technologies to inactivate foodborne viruses.     

Phage inactivation of foodborne pathogens on cooked and raw meat

Phages infecting Salmonella Typhimurium PT160 and Campylobacter jejuni were added at a low or high (10 or 10(4)) multiplicity of infection (MOI) to either low or high (<100 or 10(4)cm(-2)) densities of host bacteria inoculated onto raw and cooked beef, and incubated at 5 and 24 degrees C to simulate refrigerated and room temperature storage. Counts of host bacteria were made throughout the incubation period, with phages being counted at the first and last sampling times. Host inactivation was variable and depended on the incubation conditions and food type. Significant host inactivations of the order of 2-3 log(10)cm(-2) at 5 degrees C and >5.9 log(10)cm(-2) at 24 degrees C were achieved compared to phage-free controls using the Salmonella phage under optimal conditions (high host cell density and MOI). These results alongside those already published indicate that phages may be useful in the control for foodborne pathogens.     

Atmospheric plasma inactivation of foodborne pathogens on fresh produce surfaces

A study was conducted to determine the effect of one atmosphere uniform glow discharge plasma (OAUGDP) on inactivation of Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes on apples, cantaloupe, and lettuce, respectively. A five-strain mixture of cultured test organisms was washed, suspended in phosphate buffer, and spot inoculated onto produce (7 log CFU per sample). Samples were exposed inside a chamber affixed to the OAUGDP blower unit operated at a power of 9 kV and frequency of 6 kHz. This configuration allows the sample to be placed outside of the plasma generation unit while allowing airflow to carry the antimicrobial active species, including ozone and nitric oxide, onto the food sample. Cantaloupe and lettuce samples were exposed for 1, 3, and 5 min, while apple samples were exposed for 30 s, 1 min, and 2 min. After exposure, samples were pummeled in 0.1% peptone water-2% Tween 80, diluted, and plated in duplicate onto selective media and tryptic soy agar and incubated as follows: E. coli O157:H7 (modified eosin methylene blue) and Salmonella (xylose lysine tergitol-4) for 48 h at 37 degrees C, and L. monocytogenes (modified Oxford medium) at 48 h for 32 degrees C. E. coli O157:H7 populations were reduced by >1 log after 30-s and 1-min exposures and >2 log after a 2-min exposure. Salmonella populations were reduced by >2 log after 1 min. Three- and 5-min exposure times resulted in >3-log reduction. L. monocytogenes populations were reduced by 1 log after 1 min of exposure. Three- and 5-min exposure times resulted in >3- and >5-log reductions, respectively. This process has the capability of serving as a novel, nonthermal processing technology to be used for reducing microbial populations on produce surfaces.     

食源性细菌低温适应的分子机制研究进展

冷藏可抑制食源性细菌的生长,延长食品的货架期,是最为常用的食品保藏策略。然而,许多食源性致病菌和腐败菌具有较强的低温适应能力,这严重威胁到食品的质量与安全。阐明食源性细菌低温适应的分子机制,对制定和实施更有效的控制措施是至关重要的。主要从食源性细菌低温应激响应关键基因的发掘,转录组学和蛋白组学在食源性细菌低温适应机制研究中的应用两个方面展开综述,以期为食源性细菌的研究和控制提供理论参考。     

食源性致病菌快速检测方法研究进展

食源性致病菌是影响食品安全的重要因素,随着生物技术发展及人们对食品安全要求的提高,食源性致病菌的检测方法也不断更新和升级。微流控技术将样品预处理、分离、检测等过程集成在微小的芯片上完成多种功能,而液滴微流控作为其中一个重要的分支,可利用两液体互不相溶特性形成的分散微液滴进行高通量的测试。本文针对核酸扩增方式的不同,比较总结了液滴微流控-数字化聚合酶链式反应、液滴微流控-数字化环介导等温扩增、液滴微流控-数字化重组酶聚合酶扩增检测方法的特点,介绍了其在食源性致病菌检测中的应用,并对该技术的未来发展前景进行了展望。液滴微流控技术结合数字化核酸扩增,可实现对食源性致病菌的智能化、连续化、精准化和微型化的快速检测,本文为其在食源性致病菌快速检测领域的发展提供了参考资料。     

Thermal and high-pressure inactivation kinetics of carrot pectinmethylesterase: from model system to real foods

Pectinmethylesterase (PME) was extracted from carrots (Daucus carrota) and subsequently purified by affinity chromatography on a CNBr-Sheparose-PME inhibitor (PMEI) column. Detailed kinetic studies on the inactivation of purified carrot PME by thermal and high-pressure processing were performed. Next to the model systems, the thermal and high-pressure inactivation of PME in real products (carrot juice and carrot pieces) was investigated. The inactivation was performed under isothermal and isothermal–isobaric conditions. Under all conditions investigated, a first-order kinetic model could describe the inactivation data. It was found that carrot PME is much more thermostable and pressure-stable in carrot pieces than in carrot juice or purified form.     

奶粉中病原菌质控考核结果分析

目的 对质控考核样品进行检测, 分离鉴定肠杆菌科志贺氏菌属A群痢疾志贺氏菌和金黄色葡萄球菌, 并分析其中特别的生物学现象。方法 按照《食品安全国家标准 食品微生物学检验 (GB 4789.5-2012)》对考核检样进行检测。结果 分离出宋内志贺氏菌而未分离出金黄色葡萄球菌, 并在检验过程中观察到特别的生物学现象。结论 志贺氏菌在志贺氏菌属显色培养基上并非显示一样颜色, 有白色菌落周围培养基显示紫红色,也有黄色菌落周围培养基显示黄色。     

纳米材料在食源性致病菌检测中的应用

食品安全问题是国内外一直都密切关注的热点问题,与每个人的生活都息息相关,而食源性致病菌则是引发食品安全问题的主要因素之一。快速、准确的检测食源性致病菌是控制这类问题的关键所在。传统的检测方法存在耗时长,特异性和灵敏性较差等问题。而将纳米材料与传统食源性致病菌检测方法相结合,能够有效解决此类问题,极大程度地促进了致病菌检测的研究进展。文章就几种常见的纳米材料在食源性致病菌检测中的应用进行了综合评述并对其未来的研究方向进行了展望。     

食源性致病菌快速检测方法的探索研究

目的探讨传统细菌培养法及荧光定量PCR检测两种检测方法在食源性致病菌上的检出效果。方法对2015年采集的7类食品共600例样品采用传统细菌培养法及增菌后的增菌液荧光定量PCR检测,对检测结果进行分析比较。结果被检食品存在食源性致病菌污染,600例样品共检测出35株致病菌,食品致病菌的总检出率为5.83%,其中副溶血性弧菌与金黄色葡萄球菌检出率最高,达到15%。蜡样芽胞杆菌检出率较高,达到10.00%。传统细菌培养法致病菌阳性检出率为2.67%,荧光定量PCR检测致病菌阳性检出率为3.17%,两种检测方法下的检出率差异没有明显差异(X~2=1.882,P0.05)。结论检测中将传统细菌培养法同荧光定量PCR检测这两种检测方法结合起来,不仅提高了食源性致病菌检出率,也可缩短检测时间,提高检出率。后续研究将继续加大样本量并覆盖更多食品种类。     

食源性致病菌RPA检测技术研究进展

食源性致病菌是食品安全的重要隐患,开发针对食源性致病菌快速、有效的检测技术迫在眉睫。在诸多食源性致病菌检测手段中,重组酶聚合酶扩增技术(RPA),因具有比传统分子及免疫学检测技术如聚合酶链式反应(PCR)、实时荧光定量PCR、酶联免疫吸附技术(ELISA)等更快速、灵敏、特异、简便以及实用性强等方面优势,现已在一定程度上得到应用。本文将对RPA技术原理及其衍生技术包括直接重组酶聚合酶扩增技术(Direct-RPA)、重组酶聚合酶扩增侧流层析技术(RPA-LFD)、重组酶聚合酶扩增酶联免疫吸附技术(RPA-ELISA)、实时重组酶聚合酶扩增技术(Real-time RPA)、RPA微流体等技术进行简要综述。     

竹叶提取物对食源性细菌抑制效应的研究

目的 利用两种方法对竹叶有效抑菌成分的进行提取,并对3种食源性微生物进行抑菌圈试验比较, 研究竹叶提取物对食源性细菌的生长抑制效果以及对比两种提取方法的不同提取效果。方法 利用乙醇、乙酸乙酯、丙酮等有机溶剂为提取介质, 使用水浴浸提法和超声波微波提取法获得竹叶提取液。并利用提取物对大肠杆菌、金黄色葡萄球菌、恶臭假单胞菌三种目标菌株使用抑菌圈法进行抑菌实验。结果 利用乙醇 冰醋酸、丙酮、乙酸乙酯为提取介质的抑菌圈实验表明: 三种提取介质都能提取到竹叶有效抑菌成分, 但乙酸乙酯的提取效果相对较好, 另外, 超声波微波提取法在提取时间和效果上均优于水浴浸提法。结论 竹叶提取液对食源性细菌的生长有较高的抑制作用, 可用于天然食品防腐剂的开发。在竹叶有效成分的提取方面超声波微波提取法可替代传统的水浴浸提法。     

食源性致病菌高光谱识别模型构建

针对食源性致病菌检测过程耗时费力的问题,应用高光谱成像技术获取了12,18,24h3个生长期的5类致病菌[金黄色葡萄球菌(S.aureus)、单增李斯特菌(LM)、致泻大肠埃希氏菌(DEC)、肠炎沙门氏菌(SE)和福氏志贺菌(S.flexneri)]的高光谱图像,利用连续投影算法(SPA)结合相关分析(CA)提取不同生长期致病菌敏感波段并建立了最小二乘支持向量机(LS-SVM)判别模型。结果表明,5类致病菌对不同波长光的反射能力不同。SPA-CA筛选出的5个敏感波段(462,498,649,853,979nm)的反射率可较好地反映5类致病菌在不同生长期的波谱特性。基于该敏感波段反射率构建的LS-SVM模型能有效地判别出S.aureus、LM和DEC 3类致病菌,而SE和S.flexneri两种属容易被互相误判,SE被误判成S.flexneri的概率为11.2%,S.flexneri被误判成SE的概率为19.9%。LS-SVM模型对5类致病菌的总体识别正确率为90.9%。综上,高光谱成像技术结合化学计量学方法具有快速诊断食源性致病菌的能力。     

Enhanced inactivation of foodborne pathogenic and spoilage bacteria by FD&C Red no. 3 and other xanthene derivatives during ultrahigh pressure processing

Variability among microorganisms in barotolerance has been demonstrated at genus, species, and strain levels. Identification of conditions and additives that enhance the efficacy of ultrahigh pressure (UHP) against important foodborne microorganisms is crucial for maximizing product safety and stability. Preliminary work indicated that FD&C Red No. 3 (Red 3), a xanthene derivative, was bactericidal and acted synergistically with UHP against Lactobacillus spp. The objective of this study was to determine the antimicrobial efficacy of Red 3 and other xanthene derivatives, alone and combined with UHP, against spoilage and pathogenic bacteria in citrate-phosphate buffer (pH 7.0). Xanthene derivatives tested were fluorescein, Eosin Y, Erythrosin B, Phloxine B, Red 3, and Rose Bengal. Halogenated xanthene derivatives (10 ppm) were effective at reducing Listeria monocytogenes survivors but ineffective against Escherichia coli O157:H7. When combined with UHP (400 MPa, 3 min), the presence of derivatives enhanced inactivation. Because Red 3 was the only xanthene derivative to produce synergistic inactivation of both pathogens, further studies using this colorant were warranted. Efficacy of Red 3 against gram-positive bacteria (Lactobacillus plantarum and L. monocytogenes) was concentration dependent (1 to 10 ppm). E. coi O157: H7 strains were resistant to Red 3 concentrations up to 300 ppm. When Red 3 was combined with UHP, the lethality against gram-positive and gram-negative bacteria was dose dependent, with synergy being significant for most strains at > or = 3 ppm. Additional gram-positive and gram-negative bacteria showed lethalities similar to those observed for L. plantarum or L. monocytogenes, and E. coli O157:H7, respectively. Red 3 is a potentially useful additive to enhance the safety and stability of UHP-treated food products.