Inactivation of biofilm cells of foodborne pathogen by aerosolized sanitizers

The objective of this study was to determine the effect of aerosolized sanitizers on the inactivation of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes biofilms. Biofilms were formed on a stainless steel and polyvinyl chloride (PVC) coupon by using a mixture of three strains each of three foodborne pathogens. Six day old biofilms on stainless steel and PVC coupons were treated with aerosolized sodium hypochlorite (SHC; 100 ppm) and peracetic acid (100, 200, and 400 ppm) in a model cabinet for 5, 10, 30, and 50 min. Treatment with 100 ppm PAA was more effective than the same concentration of SHC with increasing treatment time. Exposure to 100 ppm SHC and PAA for 50 min significantly (p<0.05) reduced biofilm cells of three foodborne pathogens (0.50 to 3.63 log CFU/coupon and 2.83 to more than 5.78 log CFU/coupon, respectively) compared to the control treatment. Exposure to 200 and 400 ppm PAA was more effective in reducing biofilm cells. Biofilm cells were reduced to below the detection limit (1.48 log CFU/coupon) between 10 and 30 min of exposure. The results of this study suggest that aerosolized sanitizers have a potential as a biofilm control method in the food industry.     

Effect of NaCl on the biofilm formation by foodborne pathogens

This study was designed to evaluate the effect of NaCl on the biofilm formation of Listeria monocytogenes, Staphylococcus aureus, Shigella boydii, and Salmonella Typhimurium. The biofilm cells were cultured in media containing different NaCl concentrations (0% to 10%) for 10 d of incubation at 37 °C using a 24-well polystyrene microtiter plate, collected by swabbing methods, and enumerated using plate count method. The attachment and detachment kinetic patterns were estimated according to the modified Gompertz model. The cell surface hydrophobicity and auto-aggregation were observed at different NaCl concentrations. Most strains showed 2 distinctive phases at lower than 6% NaCl, while the numbers of adhered cells gradually increased throughout the incubation period at 4% to 10% NaCl. At 0% NaCl, the numbers of adhered L. monocytogenes, S. aureus, S. boydii, and S. Typhimurium cells rapidly increased up to 7.04, 6.47, 6.39, and 7.27 log CFU/cm(2), respectively, within 4 d of incubation. The maximum growth rate (k(A)) and specific growth rate (μ(A)) of adherent pathogenic cells were decreased with increasing NaCl concentration. Noticeable decline in the numbers of adherent cells was observed at low concentration levels of NaCl (<2%). The adherence abilities of foodborne pathogens were influenced by the physicochemical surface properties. The hydrophobicity and auto-aggregation enhanced the biofilm formation during the incubation periods. Therefore, this study could provide useful information to better understand the adhesion and detachment capability of foodborne pathogens on food contact surfaces.     

Inactivation of foodborne pathogens in ready-to-eat salad using UV-C irradiation

To examine the applicability of ultraviolet (UV)-C irradiation on the inactivation of foodborne pathogen in ready-to-eat salad, it was inoculated with Escherichia coli O157:H7 and Listeria monocytogenes and then irradiated with UV-C light. Radiation dose required for 90% reduction (d _R) values of E. coli O157:H7 and L. monocytogenes were determined to be 0.21 and 2.48 J/m², respectively. Foodborne pathogen populations significantly (p<0.05) decreased with increasing UV-C irradiation. UV-C irradiation at 8,000 J/m² reduced the populations of E. coli O157:H7 and L. monocytogenes on ready-to-eat salad by 2.16 and 2.57 log CFU/g, respectively.     

Inactivation of foodborne pathogens in milk using dynamic high pressure

Improving the microbiological safety of perishable foods is currently a major preoccupation in the food industry. The aim of this study was to investigate the inactivation of three major food pathogens (Listeria monocytogenes [LSD 105-1], Escherichia coli O157:H7 [ATCC 35150], and Salmonella enterica serotype Enteritidis ATCC [13047]) by dynamic high pressure (DHP) in order to evaluate its potential as a new alternative for the cold pasteurization of milk. The effectiveness of DHP treatment against L. monocYtogenes, E. coli O157:H7, and Salmonella Enteritidis was first evaluated in 0.01 M phosphate-buffered saline (PBS) at pH 7.2 as a function of applied pressure (100, 200, and 300 MPa) and of the number of passes (1, 3, and 5) at 25 degrees C. A single pass at 100 MPa produced no significant inactivation of the three pathogens, while increasing the pressure up to 300 MPa or the number of passes to five increased inactivation. From an initial count of 8.3 log CFU/ml, complete inactivation of viable L. monocytogenes was achieved after three successive passes at 300 MPa, while 200-MPa treatments with three and five passes completely eliminated viable Salmonella Enteritidis and E. coli O157:H7, respectively. The effectiveness of DHP for the inactivation of these pathogens was compared to that of hydrostatic high pressure (HHP) using the same pressure (200 MPa, single pass at 25 degrees C). In general, two additional log reductions in viable count were obtained with DHP DHP was less effective against L. monocytogenes and E. coli O157:H7 in raw milk than in PBS. After five passes at 200 MPa, an 8.3-log reduction was obtained for E. coli O157:H7, while a reduction of about 5.8 log CFU/ml was obtained for L. monocytogenes exposed to 300 MPa for five passes. Exposing milk or buffer samples to mild heating (45 to 60 degrees C) prior to dynamic pressurization enhanced the lethal effect of DHP The inactivation of pathogens also depended on the initial bacterial concentration. The highest reduction was obtained when the bacterial load did not exceed 10(5) CFU/ml. In conclusion, DHP was shown to be very effective for the destruction of the tested pathogens. It offers a promising alternative for the cold pasteurization of milk and possibly other liquid foods.     

Emerging foodborne pathogens

The broad spectrum of foodborne infections has changed dramatically over time, as well-established pathogens have been controlled or eliminated, and new ones have emerged. The burden of foodborne disease remains substantial: one in four Americans is estimated to have a significant foodborne illness each year. The majority of these illnesses are not accounted for by known pathogens, so more must remain to be discovered. Among the known foodborne pathogens, those more recently identified predominate, suggesting that as more and more is learned about pathogens, they come under control. In addition to the emergence or recognition of new pathogens, other trends include global pandemics of some foodborne pathogens, the emergence of antimicrobial resistance, the identification of pathogens that are highly opportunistic, affecting only the most high-risk subpopulations, and the increasing identification of large and dispersed outbreaks. New pathogens can emerge because of changing ecology or changing technology that connects a potential pathogen with the food chain. They also can emerge de novo by transfer of mobile virulence factors, often through bacteriophage. Though this is rarely observed, it can be reconstructed. Better understanding of the ecology and dynamics of phage transmission among bacteria will help us to understand the appearance of new pathogens in the future. One may look for emerging foodborne pathogens among the silent zoonoses, and among the severe infections affecting the immunocompromised humans. We should expect the unexpected. In the past, separating human sewage and animal manure from human food and water supplies was critical to improving public health. Now, our health depends increasingly on the safety of the feed and water supplies for the animals themselves. The successes of the 20th century and the new challenges we face mean that public health vigilance, careful investigation of new problems, responsible attention to food safety from farm to table, and partnerships to bring about new foodborne disease control measures will be needed for the foreseeable future.     

Inactivation of foodborne pathogens by the synergistic combinations of food processing technologies and food‐grade compounds

There is a need to develop food processing technologies with enhanced antimicrobial capacity against foodborne pathogens. While considering the challenges of adequate inactivation of pathogenic microorganisms in different food matrices, the emerging technologies are also expected to be sustainable and have a minimum impact on food quality and nutrients. Synergistic combinations of food processing technologies and food‐grade compounds have a great potential to address these needs. During these combined treatments, food processes directly or indirectly interact with added chemicals, intensifying the overall antimicrobial effect. This review provides an overview of the combinations of different thermal or nonthermal processes with a variety of food‐grade compounds that show synergistic antimicrobial effect against pathogenic microorganisms in foods and model systems. Further, we summarize the underlying mechanisms for representative combined treatments that are responsible for the enhanced microbial inactivation. Finally, regulatory issues and challenges for further development and technical transfer of these new approaches at the industrial level are also discussed.     

Inactivation of Cryptosporidium parvum oocysts in cider by flash pasteurization

Cryptosporidium parvum is a well-recognized pathogen of significant medical importance, and cider (apple juice) has been associated with foodborne cryptosporidiosis. This study investigated the effect of flash pasteurization on the viability of contaminant C. parvum oocysts. Cider inoculated with oocysts was heated at 70 or 71.7 degrees C for 5, 10, or 20 s, and oocyst viability was measured by a semiquantitative in vitro infectivity assay. By infecting multiple wells of confluent Madin-Darby bovine kidney cells with serial dilutions of heat-treated oocysts and examining infected cells by indirect fluorescent antibody staining, the most probable number technique was applied to quantify log reduction of oocyst viability. Heating for 10 or 20 s at either temperature caused oocyst killing of at least 4.9 log (or 99.999%), whereas oocyst inactivation after pasteurization for 5 s at 70 and 71.7 degrees C was 3.0 log (99.9%) and 4.8 log (99.998%), respectively. Our results suggested that current practices of flash pasteurization in the juice industry are sufficient in inactivating contaminant oocysts.     

Inhibition of biofilm formation,adhesion and invasion in Caco-2 cells by foodborne pathogens using phyto-mediated synthesised silver nanoparticles from industrial wastes

Certain foodborne diseases are associated with antibiotic resistance, a significant problem throughout the world. Silver nanoparticles (AgNPs) using industrial waste from Eucalyptus camaldulensis and sericin, a protein derived from Bombyx mori, were synthesised by a one-step approach. Spherical-shaped nanoparticles with the average size of 17.19 nm exhibited strong antioxidant activity. The minimum bactericidal concentrations against foodborne pathogens including Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus, Escherichia coli O157:H7, Klebsiella pneumoniae, Salmonella Typhimurium, Shigella sonnei, Vibrio cholerae and Vibrio parahaemolyticus were between 2.96 and 11.83 µg/mL. Killing against L. monocytogenes and E. coli O157:H7 was observed within 4 h. Treatment with AgNPs at 0.25 – 0.5 × MIC significantly reduced biofilm production in all isolates (P < 0.05). AgNPs significantly impeded adhesion to and invasion of human epithelial Caco-2 cells by L. monocytogenes and E. coli O157:H7 (P < 0.05). Biocompatibility assessment of AgNPs with Caco-2 and human red blood cells demonstrated no toxic effects.     

Growth inhibition of foodborne pathogens by Oenococcus oeni

Abstract: To explore the possibility of using Oenococcus oeni to inhibit foodborne pathogens, and to characterize antimicrobial compounds produced by O. oeni, 24 strains of O. oeni were tested for their ability to inhibit growth of foodborne pathogens, Escherichia coli O157:H7, Salmonella enteritidis, and Listeria monocytogenes by using the spot‐on‐lawn method. Of the 24 strains, 17 strains were able to inhibit all 3 pathogens in this study. Proteases, catalase, and buffer solutions were used for determining the type of inhibitory compounds produced from 4 selected strains with stronger inhibitory activity. Antimicrobial activity of 2 strains against the pathogens was completely inactivated by buffer solution, and other 2 strains against E. coli O157:H7 were partially removed. The antimicrobial compound was not sensitive to selected proteases and catalase. Practical Application: There is little information available about using O. oeni for human pathogens control. The results of this study revealed such discovery and potential applications for pathogen control.     

Pore-forming toxins of foodborne pathogens

Pore-forming toxins (PFTs) are water-soluble molecules that have been identified as the most crucial virulence factors during bacterial pathogenesis. PFTs disrupt the host cell membrane to internalize or to deliver other bacterial or virulence factors for establishing infections. Disruption of the host cell membrane by PFTs can lead to uncontrollable exchanges between the extracellular and the intracellular matrix, thereby disturbing the cellular homeostasis. Recent studies have provided insights into the molecular mechanism of PFTs during pathogenesis. Evidence also suggests the activation of several signal transduction pathways in the host cell on recognition of PFTs. Additionally, numerous distinctive host defense mechanisms as well as membrane repair mechanisms have been reported; however, studies reveal that PFTs aid in host immune evasion of the bacteria through numerous pathways. PFTs have been primarily associated with foodborne pathogens. Infection and death from diseases by consuming contaminated food are a constant threat to public health worldwide, affecting socioeconomic development. Moreover, the emergence of new foodborne pathogens has led to the rise of bacterial antimicrobial resistance affecting the population. Hence, this review focuses on the role of PFTs secreted by foodborne pathogens. The review highlights the molecular mechanism of foodborne bacterial PFTs, assisting bacterial survival from the host immune responses and understanding the downstream mechanism in the activation of various signaling pathways in the host upon PFT recognition. PFT research is a remarkable and an important field for exploring novel and broad applications of antimicrobial compounds as therapeutics.     

Molecular-genetic analysis of foodborne pathogens

The review of the modern foreign and national publications on application of modern molecular and DNA-based analysis in food microbiology is given. Most frequently used nucleonic acids-based assays for foodborne pathogens, including DNA-hybridization, conventional PCR and real-time PCR are listed. The basic advantages and lacks of PCR-techniques also problems of their application are stated at the analysis of foods. Necessity of development of criteria and methodology of standardization of diagnostic PCR for the detection of foodborne pathogens is shown, with a view of harmonization of genetic methods with existing system of microbiological requirements and control.     

生物膜状态对食源性致病菌耐药与毒力的影响研究进展

生物膜是细菌抵御不利环境维持群体稳定性的一种常见的群落形态。生物膜状态增强了食源性致病菌的抗逆性和持久存活力,促进了细菌间的信息传导和物质交换。特别是生物膜状态显著增强了菌株对抗生素的耐受能力,提升了可移动元件在细菌间的转移效率,并且生物膜状态下的细菌具备更强的入侵和感染能力,成为食品安全和人类健康的重要危害。基于此,本文将从生物膜的结构特征与异质性、生物膜对基因突变和基因水平转移的影响、以及群体感应调控等多个角度,简述生物膜状态影响食源性致病菌耐药和毒力的研究进展,以期为深入研究生物膜的生物功能与危害防控提供新的思路。     

鲜榨柠檬原汁对7种常见食源性致病菌抑菌效果及被膜形成的影响

目的研究鲜榨柠檬果皮和果肉原汁对7种常见食源性致病菌的抑菌效果以及对生物被膜形成的影响。方法对柠檬果皮和果肉分别榨汁测定总酸;采用顶空取样结合气相色谱-质谱联用技术(GC-MS)分析鲜榨柠檬果皮和果肉原汁中的主要挥发性成分;采用平板抑菌试验研究两种鲜榨原汁的抑菌效果;采用试管法和96孔板法研究两种鲜榨原汁对细菌生物被膜形成的影响。结果鲜榨柠檬果肉原汁总酸为98.86 g/kg,鲜榨柠檬果皮原汁总酸为6.05 g/kg;GC-MS结果表明鲜榨柠檬果皮和果肉原汁中主要挥发性成分以柠檬烯、γ-萜品烯、α-松油醇、柠檬醛等为主;鲜榨柠檬果肉原汁对7种常见食源性致病菌均具有明显抑菌效果,鲜榨柠檬果皮原汁均未出现明显抑菌作用;鲜榨柠檬果肉原汁对部分细菌被膜形成有抑制作用,鲜榨柠檬果皮原汁对部分细菌被膜形成有明显促进作用。结论鲜榨柠檬原汁对常见食源性致病菌生长及被膜形成影响的研究为食源性致病菌的防控提供了科学依据。     

多重PCR检测4种常见食源性致病菌

目的 建立一种多重聚合酶链式反应法(multiplex polymerase chain reaction, MPCR)快速检测肉制品中金黄色葡萄球菌、沙门氏菌、志贺氏菌和单增李斯特氏菌的分析方法。方法 选取金黄色葡萄球菌nuc基因、沙门氏菌SipB基因、志贺氏菌ipaH基因、单增李斯特菌inlA基因作为目标基因, 设计4对PCR引物, 建立并优化多重PCR反应体系, 评价该体系的特异性和灵敏度, 并对人工污染的熟肉样品进行检测。结果 构建的多重PCR方法特异性强、灵敏度高, 人工污染熟肉匀浆中4种致病菌的检出限为103 CFU/mL。结论 构建的多重PCR检测方法能够快速、准确、高效地检测肉制品中金黄色葡萄球菌、沙门氏菌、志贺氏菌和单增李斯特氏菌, 为食源性疾病菌的快速检测提供参考依据。     

Inactivation effect of dielectric barrier discharge plasma against foodborne pathogens on the surfaces of different packaging materials

The usefulness of dielectric barrier discharge (DBD) plasma for surface disinfection of the common food packaging materials, namely glass, polyethylene, polypropylene, nylon, and paper foil was evaluated. DBD plasma was generated by applying a pulsed DC voltage of 10 kV and at a power of 208 W. The separation distance between the electrodes was 2.65 mm. On exposure of food pathogens-loaded packaging materials to the plasma, > 4 log/cm² reduction (99.99%) in viable cell counts of Escherichia coli O157:H7 was observed in 10 min. The other two tested pathogen strains, Salmonella typhimurium and Staphylococcus aureus, were inactivated in the range 3.0–3.5 log/cm². The inactivation pattern of the pathogens fitted well to the log-linear and tail model. Compared to unexposed packaging materials, no significant (p > 0.05) changes in the surface temperatures, optical characteristics, tensile strengths, and strain-induced deformation were observed for the DBD plasma-exposed materials. Therefore, the DBD plasma can be used to disinfect surfaces of different food packaging materials harboring moderate levels of bacterial contaminants without adversely affecting their physicomechanical properties.Industrial relevanceTraditionally, dry heat, steam, UV light and chemicals like ethylene oxide and hydrogen peroxide have been used as surface sterilants and disinfectants for packaging materials in the food industry. However, certain limitations have motivated the search for new approaches. Cold plasma technology is an emerging, green process for surface sterilization. The DBD plasma was found to be effective in reducing the bacterial food pathogens on different food packaging materials. As the technology is simple and scalable, it can be readily applied industrially.     

Inactivation of pathogens on pork by steam-ultrasound treatment

The objective of the study was to evaluate a new pathogen inactivation concept that combines application of pressurized steam simultaneously with high-power ultrasound through a series of nozzles. On skin and meat surfaces of pork jowl samples, counts of total viable bacteria were reduced by 1.1 log CFU/cm(2) after treatment for 1 s and by 3.3 log CFU/cm(2) after treatment for 4 s. The mean reduction of 1.7 to 3.3 log CFU/cm(2) on the skin surface was significantly higher than the reduction of 1.1 to 2.5 log CFU/cm(2) on the meat surface. The inactivation of Salmonella Typhimurium, Salmonella Derby, Salmonella Infantis, Yersinia enterocolitica, and a nonpathogenic Escherichia coli was studied on inoculated samples that were treated for 0.5 to 2.0 s. With one exception, no significant differences in reduction were observed among the bacterial types. After treatment for 0.5 s, the 0.9-to 1.5-log reductions of E. coli were significantly higher than the 0.4- to 1.1-log reductions for Salmonella and Y. enterocolitica. Overall, reductions increased by increasing treatment time; reductions were 0.4 to 1.5 log CFU/cm(2) after treatment for 0.5 s and 2.0 to 3.6 log CFU/cm(2) after treatment for 2 s. Reductions on the skin (1 to 3.6 log CFU/cm(2)) were significantly higher than reductions on the meat surface (1 to 2.5 log CFU/cm(2)). The reduced effect on the meat surface may be explained by greater protection of bacteria in deep structures at the muscle surface. No significant difference in reduction was observed between samples inoculated with 10(4) CFU/cm(2) and those inoculated with 10(7) CFU/cm(2), and cold storage of samples for 24 h at 5°C after steam-ultrasound treatment did not lead to changes in recovery of bacteria.     

Exposure to 222 nm far UV-C effectively inactivates planktonic foodborne pathogens and inhibits biofilm formation

This study investigated the performance of a 222 nm far-UV-C krypton-chloride excilamp for inactivation of major foodborne pathogenic and spoilage bacteria in thin liquid films (TLF, 1.2 mm thickness), on solid stainless steel surfaces (SS), and against biofilm formation on SS. Both gram-positives (Listeria monocytogenes, Staphylococcus aureus) and gram-negatives (Escherichia coli O157:H7, Pseudomonas aeruginosa) (10⁹ CFU/mL starting concentration) were exposed to 222 nm light at cumulative doses of up to 354 mJ/cm². Significant (P < 0.05) reductions (1.4–5.1 log CFU) were found for all bacteria, and inactivation kinetics was described well by the Weibull model (0.77 ≤ R² ≤ 0.95). Substrate type (i.e., TLF vs. SS) substantially impacted treatment efficacy. No detectable resistance of L. monocytogenes was developed after repeated exposure to 222 nm in TLF. The 222 nm treatment also effectively minimized biofilm formation and growth by S. aureus and P. aeruginosa and increased the surviving cells' susceptibility to sodium hypochlorite by at least 2 fold.Industrial relevanceThis work demonstrates that 222 nm krypton-chloride excilamps can be used to effectively inactivate planktonic bacteria and inhibit biofilm formation and growth. This recommends them for use as novel nonthermal light-based systems for mitigation of pathogens and biofilms in a range of applications, including food processing, food service, and clinical environments.     

Microbiology and foodborne pathogens in honey

Honey has been considered a relatively safe foodstuff due to its compositional properties, with infant botulism caused by Clostridium botulinum being the most prominent health risk associated with it. Our review is focused on the honey microflora along the food chain and evaluates the pathogenic potential of those microorganisms found in honey. This product may contain a great variety of bacteria and, particularly, fungi that eventually entered the food chain at an early stage (e.g., via pollen). For many of these microorganisms, opportunistic infections in humans have been recorded (e.g., infections by Staphylococcus spp., Citrobacter spp., Escherichia coli, Hafnia alvei, Aspergillus spp., Fusarium spp., Trichoderma spp., Chaetomium spp.), although direct infections via honey were not registered.