Selection Criteria for Water Disinfection Techniques in Agricultural Practices

This paper comprises a selection tool for water disinfection methods for fresh produce pre- and postharvest practices. A variety of water disinfection technologies is available on the market and no single technology is the best choice for all applications. It can be difficult for end users to choose the technology that is best fit for a specific application. Therefore, the different technologies were characterized in order to identify criteria that influence the suitability of a technology for pre- or postharvest applications. Introduced criteria were divided into three principal components: (i) criteria related to the technology and which relate to the disinfection efficiency, (ii) attention points for the management and proper operation, and (iii) necessities in order to sustain the operation with respect to the environment. The selection criteria may help the end user of the water disinfection technology to obtain a systematic insight into all relevant aspects to be considered for preliminary decision making on which technologies should be put to feasibility testing for water disinfection in pre- and postharvest practices of the fresh produce chain.     

Resistance of Listeria monocytogenes, Escherichia coli O157:H7 and Campylobacter jejuni after exposure to repetitive cycles of mild bactericidal treatments

While maintaining nutritional and sensorial attributes of fresh foods mild processing technologies generally deliver microbiologically perishable food products. Currently little information exists on possible increase in the resistance of pathogens after repetitive exposure to mild (sub-lethal) treatments. Multiple strain-cocktails of Listeria monocytogenes, Escherichia coli O157:H7 and Campylobacter jejuni were exposed to 20 consecutive cycles of sub-lethal inactivation by three different techniques. Used techniques comprised inactivation with lactic acid (LA), chlorine dioxide (ClO₂) and intense light pulses (ILP). Results showed that the selection of resistant cells was both species and technique dependent. While repetitive cycles of ClO₂ treatment did not result in increased resistance, repetitive inactivation with LA yielded L. monocytogenes culture of higher resistance in comparison to the parental culture. The increased resistance, expressed as decreased level of reduction in bacterial counts in subsequent inactivation cycles, was also observed with ILP for both L. monocytogenes and E. coli O157:H7 strains. Visual trend observations were confirmed through statistical linear regression analysis. No such effects were noted for C. jejuni which became undetectable after first 2–5 cycles. Current findings indicate the ability of foodborne pathogens to adapt to mild bactericidal treatments creating new challenges in risk assessment and more specifically in hazard analysis.     

Zero Risk Does Not Exist: Lessons Learned from Microbial Risk Assessment Related to Use of Water and Safety of Fresh Produce

Risk assessments related to use of water and safety of fresh produce originate from both water and food microbiology studies. Although the set‐up and methodology of risk assessment in these 2 disciplines may differ, analysis of the current literature reveals some common outcomes. Most of these studies from the water perspective focus on enteric virus risks, largely because of their anticipated high concentrations in untreated wastewater and their resistance to common wastewater treatments. Risk assessment studies from the food perspective, instead, focus mainly on bacterial pathogens such as Salmonella and pathogenic Escherichia coli. Few site‐specific data points were available for most of these microbial risk assessments, meaning that many assumptions were necessary, which are repeated in many studies. Specific parameters lacking hard data included rates of pathogen transfer from irrigation water to crops, pathogen penetration, and survival in or on food crops. Data on these factors have been investigated over the last decade and this should improve the reliability of future microbial risk estimates. However, the sheer number of different foodstuffs and pathogens, combined with water sources and irrigation practices, means that developing risk models that can span the breadth of fresh produce safety will be a considerable challenge. The new approach using microbial risk assessment is objective and evidence‐based and leads to more flexibility and enables more tailored risk management practices and guidelines. Drawbacks are, however, capacity and knowledge to perform the microbial risk assessment and the need for data and preferably data of the specific region.     

Antimicrobial effect of spices and herbs on Shigella sonnei and Shigella flexneri

Of 17 spices and herbs tested at 1% (wt/vol) in Mueller-Hinton (MH) agar, only cloves, thyme, oregano, allspice, basil, rosemary, and marjoram showed antimicrobial effects on Shigella. The MICs of thyme, oregano, basil, and rosemary (as determined by the agar dilution method) ranged from 0.5 to 1% (wt/vol) depending on the Shigella strain used. With the use of various combinations of temperatures (12, 22, and 37 degrees C), pHs (5.0, 5.5, and 6.0), and NaCl concentrations (1, 2, 3, and 4%, wt/vol) and the inclusion or exclusion of thyme or basil at 1% (wt/vol) in an MH agar model system, it was established that basil or thyme can contribute to combination processing as a growth-inhibitory factor for Shigella spp. In the presence of basil and thyme, Shigella flexneri did not develop CFU during the 7-day incubation period for, respectively, 14 and 16 of the 18 tested combinations, while growth was noted in the corresponding temperature-pH-NaCl concentration combinations without basil or thyme. A growth-inhibitory effect on Shigella sonnei was also noted. The results of an orientation study involving the addition of basil and thyme to spaghetti sauce prior to autoclaving and S. sonnei inoculation indicated that basil and thyme contributed to the reduction of S. sonnei after 16 days at 12 degrees C but not at 4 degrees C.     

Computer aided boar semen motility analysis for cereulide detection in different food matrices

Computer Aided Semen Analysis (CASA) study of the boar semen motility has been demonstrated to be an appropriate assay for detection of cereulide (Bacillus cereus emetic toxin). Application of the boar semen bio-assay to detect cereulide directly in foods requires investigation of potential interference of food components, preservatives and other microbial and chemical food contaminants with the bio-assay. Current study provides evidence that none of included Staphylococcus aureus enterotoxins A, B, C and D nor B. cereus Hemolysin BL (HBL) and non-hemolytic enterotoxin (NHE) and three mycotoxins (Sterigmatocystin, Fumonisin B1 and Patulin) exhibited a toxic impact on semen progressive motility. Aflatoxin M1, M3 and zearalenone impaired semen motility only at concentrations (0.004 mg ml(-1), 0.1 mg ml(-1) and 10 mg ml(-1), respectively) much higher than those found in foods and those permitted by legislation, in comparison to cereulide which induces motility cease at concentrations lower than 20 ng ml(-1). Ten commonly used preservatives, namely potassium sorbate, sodium benzoate, (DL) malic acid, citric acid, (L+) tartaric acid, acetic acid, (DL) lactic acid, (L+) ascorbic acid, sodium chloride and sucrose induced no cease in spermatozoa motility even at preservative concentrations higher than permitted by legislation. Dioxins, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and acrylamide had no acute effect on spermatozoa motility at concentrations of 500 and 10,000 mg ml(-1), respectively. Robustness of computer aided boar semen motility analysis, tested with 14 different foods inoculated with cereulide producing B. cereus, showed distinct cereulide production in seven samples (although B. cereus growth to counts higher than 8 log CFU g(-1) was noted in 11 samples), in amounts close to those reported in foodborne outbreaks. Test evaluation in 33 samples suspected to hold cereulide showed actual cereulide presence in ten samples and no interference of food matrix with the assay.     

Organic acid based sanitizers and free chlorine to improve the microbial quality and shelf-life of sugar snaps

A screening in a sugar snap packaging company showed a converged build-up of aerobic psychrotrophic plate count (APC) (ca. 6.5 log CFU/100 mL), yeasts and molds (Y&M), and lactic acid bacteria (LAB) (both ca. 4.5 log CFU/100 mL) in the wash water in the absence of water sanitizer, and a low build-up of chemical oxygen demand (30 ± 5 mg O₂/L) and turbidity (5.2 ± 1.1 NTU).     

Notch4 and Wnt-1 proteins function to regulate branching morphogenesis of mammary epithelial cells in an opposing fashion

The ab initio folding problem can be divided into two sequential tasks of approximately equal computational complexity: the generation of native-like backbone folds and the positioning of side chains upon these backbones. The prediction of side-chain conformation in this context is challenging, because at best only the near-native global fold of the protein is known. To test the effect of displacements in the protein backbones on side-chain prediction for folds generated ab initio, sets of near-native backbones (< or = 4 A C alpha RMS error) for four small proteins were generated by two methods. The steric environment surrounding each residue was probed by placing the side chains in the native conformation on each of these decoys, followed by torsion-space optimization to remove steric clashes on a rigid backbone. We observe that on average 40% of the chi1 angles were displaced by 40 degrees or more, effectively setting the limits in accuracy for side-chain modeling under these conditions. Three different algorithms were subsequently used for prediction of side-chain conformation. The average prediction accuracy for the three methods was remarkably similar: 49% to 51% of the chi1 angles were predicted correctly overall (33% to 36% of the chi1+2 angles). Interestingly, when the inter-side-chain interactions were disregarded, the mean accuracy increased. A consensus approach is described, in which side-chain conformations are defined based on the most frequently predicted chi angles for a given method upon each set of near-native backbones. We find that consensus modeling, which de facto includes backbone flexibility, improves side-chain prediction: chi1 accuracy improved to 51-54% (36-42% of chi1+2). Implications of a consensus method for ab initio protein structure prediction are discussed.     

Multi-method approach indicates no presence of sub-lethally injured Listeria monocytogenes cells after mild heat treatment

Application of mild inactivation treatments follows an increasing trend in the food industry and is often combined with sub-optimal intrinsic product conditions to ensure appropriate level of microbial safety. Listeria monocytogenes was subjected to mild heat treatment (20 min at 60 degrees C) and subsequently exposed to various mild preservation conditions based on increased NaCl concentration and decreased pH. Recovery and resuscitation of L. monocytogenes cells were studied using various methods. Using 12-fold Most Probable Number (MPN) method no difference in the amount of recovered cells under adverse conditions was noted between heat-treated and non-treated L. monocytogenes cells. Time-to-detection method using on-line OD measurements showed that heat-treated L. monocytogenes cells reached detection limit faster in acidified media and NaCl supplemented media in comparison with non-heated control cells. Flow cytometry (FCM) analysis using 5-6-carboxyfluorescein diacetate (cFDA) and propidium iodide (PI) staining showed presence of low numbers of viable cells. Overall, there was no indication of sub-lethal injury in L. monocytogenes cells after mild heat treatment.