Role of Salmonella Newport cell surface structures on bacterial attachment and transfer during cucumber peeling.

Fresh cucumbers have been recognized as a vehicle in foodborne disease outbreaks since several multistate outbreaks of salmonellosis linked to fresh cucumbers occurred in the United States. Little is known about how microbial cell surface characteristics that are known to affect adhesion can influence bacterial cross-contamination and transfer. This study investigated the role of S. Newport cell surface components on bacterial attachment and transfer in cucumbers. Wild type Salmonella Newport and its transposon mutants were used to inoculate cucumbers. Attachment strength of S. Newport wild type to cucumber was not significantly different than that of mutants. Log10 percent transfer of mutant strains to edible flesh was not different from the wild type. Significantly less wild type Salmonella remained on the peel and transferred to the peeler than one mutant did, but not the other. Our results suggest that while curli and cellulose enhance Salmonella attachment to surface of cucumbers, there appear to be other mechanisms and factors that govern Salmonella transfer in cucumbers.

Efficacy of oregano and rosemary essential oils to affect morphology and membrane functions of noncultivable sessile cells of Salmonella Enteritidis 86 in biofilms formed on stainless steel.

AIMS: This study evaluated the efficacy of essential oil from Origanum vulgare L. (oregano; OVEO) and Rosmarinus officinalis L. (rosemary; ROEO) to inactivate sessile cells of Salmonella enterica serovar Enteritidis 86 (SE86) in young and mature biofilms formed on stainless steel. METHODS AND RESULTS: Ultrastructural alterations and damage in different physiological functions caused by OVEO and ROEO in noncultivable sessile cells of SE86 were investigated using scanning electron microscopy and flow cytometry. OVEO (2·5 µl ml-1 ) and ROEO (40 µl ml-1 ) were effective to eradicate young and mature biofilms formed by SE86 sessile cells on stainless steel surfaces; however, the efficacy varied with exposure time. OVEO and ROEO caused alterations in morphology of SE86 sessile cells, inducing the occurrence of bubbles or spots on cell surface. OVEO and ROEO compromised membrane polarization, permeability and efflux activity in noncultivable SE86 sessile cells. These findings show that OVEO and ROEO act by a multitarget mechanism on SE86 membrane functions. CONCLUSIONS: ROEO and OVEO showed efficacy to eradicate SE86 sessile cells in preformed biofilms on stainless steel, displaying a time-dependent effect and multitarget action mode on bacterial cell membrane. SIGNIFICANCE AND IMPACT OF THE STUDY: The study provides for the first time the effects of OVEO and ROEO on morphology and physiological functions of noncultivable sessile cells of S. Enteritidis biofilms preformed on stainless steel surfaces.

Surfactant concentration and type affects the removal of Escherichia coli from pig skin during a simulated hand wash.

The effect of surfactant type and concentration on a bland soap formulation's ability to remove bacteria from hands remains largely unstudied. Several combinations of surfactants and water were combined to test bacterial removal efficacy using a hand-washing device (two pieces of pig skin and a mechanical motor) to simulate a hand wash. A nalidixic acid-resistant, nonpathogenic strain of Escherichia coli (ATCC 11229) was used. Two anionic surfactants, sodium lauryl sulphate and sodium stearoyl lactylate, and two nonionic surfactants, poloxamer 407 and sorbitan monostearate, each in concentrations of 2, 5 and 10% were studied. A slight positive (r2  = 0·17) but significant (P = 0·03) correlation was observed between hydrophile-lipophile balance value and mean log reduction. No correlation was observed between pH of the treatment solution and the mean log reduction (r2  = 0·05, P = 0·25). A 10% sodium lauryl sulphate mixture showed the highest log reduction (x¯ = 1·1 log CFU reduction, SD = 0·54), and was the only treatment significantly different from washing with water (P = 0·0005). There was a correlation between increasing surfactant concentrations above the critical micelle concentration, and mean microbial reduction (r2  = 0·62, P = 0·001). SIGNIFICANCE AND IMPACT OF THE STUDY: This study characterizes the role of surfactants in removing microbes during a hand wash. Numerous studies address how surfactants support antimicrobial effect in soap, or cause irritation of skin, but no published studies show which surfactants are best for removing microbes. We used pig skin as a model for human skin and a lathering device to simulate a hand wash. A 10% sodium lauryl sulphate mixture was the only treatment significantly different from a water wash. There was a strong correlation between increasing surfactant concentrations above the critical micelle concentration and mean microbial reduction.

Assessing the effect of sodium dichloroisocyanurate concentration on transfer of Salmonella enterica serotype Typhimurium in wash water for production of minimally processed iceberg lettuce (Lactuca sativa L.).

UNLABELLED: This study evaluated the impact of sodium dichloroisocyanurate (5, 10, 20, 30, 40, 50 and 250 mg l(-1) ) in wash water on transfer of Salmonella Typhimurium from contaminated lettuce to wash water and then to other noncontaminated lettuces washed sequentially in the same water. Experiments were designed mimicking the conditions commonly seen in minimally processed vegetable (MPV) processing plants in Brazil. The scenarios were as follows: (1) Washing one inoculated lettuce portion in nonchlorinated water, followed by washing 10 noninoculated portions sequentially. (2) Washing one inoculated lettuce portion in chlorinated water followed by washing five noninoculated portions sequentially. (3) Washing five inoculated lettuce portions in chlorinated water sequentially, followed by washing five noninoculated portions sequentially. (4) Washing five noninoculated lettuce portions in chlorinated water sequentially, followed by washing five inoculated portions sequentially and then by washing five noninoculated portions sequentially in the same water. Salm. Typhimurium transfer from inoculated lettuce to wash water and further dissemination to noninoculated lettuces occurred when nonchlorinated water was used (scenario 1). When chlorinated water was used (scenarios 2, 3 and 4), no measurable Salm. Typhimurium transfer occurred if the sanitizer was ≥10 mg l(-1) . Use of sanitizers in correct concentrations is important to minimize the risk of microbial transfer during MPV washing. SIGNIFICANCE AND IMPACT OF THE STUDY: In this study, the impact of sodium dichloroisocyanurate in the wash water on transfer of Salmonella Typhimurium from inoculated lettuce to wash water and then to other noninoculated lettuces washed sequentially in the same water was evaluated. The use of chlorinated water, at concentration above 10 mg l(-1) , effectively prevented Salm. Typhimurium transfer under several different washing scenarios. Conversely, when nonchlorinated water was used, Salm. Typhimurium transfer occurred in up to at least 10 noninoculated batches of lettuce washed sequentially in the same water.

Observation of turbulent intermittency scaling with magnetic helicity in an MHD plasma wind tunnel.

The intermittency in turbulent magnetic field fluctuations has been observed to scale with the amount of magnetic helicity injected into a laboratory plasma. An unstable spheromak injected into the MHD wind tunnel of the Swarthmore Spheromak Experiment displays turbulent magnetic and plasma fluctuations as it relaxes into a Taylor state. The level of intermittency of this turbulence is determined by finding the flatness of the probability distribution function of increments for magnetic pickup coil fluctuations B˙(t). The intermittency increases with the injected helicity, but spectral indices are unaffected by this variation. While evidence is provided which supports the hypothesis that current sheets and reconnection sites are related to the generation of this intermittent signal, the true nature of the observed intermittency remains unknown.

Modification of turbulent transport with continuous variation of flow shear in the large plasma device.

Continuous control over azimuthal flow and shear in the edge of the Large Plasma Device (LAPD) has been achieved using a biasable limiter which has allowed a careful study of the effect of flow shear on pressure-gradient-driven turbulence and transport in the LAPD. The LAPD rotates spontaneously in the ion diamagnetic direction; positive limiter bias first reduces, then minimizes (producing a near-zero shear state), and finally reverses the flow into the electron diamagnetic direction. Degradation of particle confinement is observed in the minimum shearing state and a reduction in the turbulent particle flux is observed with increasing shearing in both flow directions. Near-complete suppression of the turbulent particle flux is observed for shearing rates comparable to the turbulent autocorrelation rate measured in the minimum shear state. Turbulent flux suppression is dominated by amplitude reduction in low-frequency (<10 kHz) density fluctuations. An increase in fluctuations for the highest shearing states is observed with the emergence of a coherent mode which does not lead to net particle transport. The variations of density fluctuations are fit well with power laws and compare favorably to simple models of shear suppression of transport.

Threshold Photoelectron Spectrum of m-Benzyne.

Due to their unusual electronic structure, the biradical m-benzyne, C6H4, and its cation are of considerable interest in chemistry. Here, the photoion mass-selected threshold photoelectron spectrum of the m-benzyne biradical is presented. An adiabatic ionization energy of 8.65 ± 0.015 eV is derived, while a vibrational progression of 0.10 eV is assigned to the ν9+ ring breathing mode, in excellent agreement with computations. The experimental spectrum was reproduced well by Franck-Condon spectral modeling of the 2A1 ← X 1A1 transition, in which the cation retains a monocyclic C6 framework. The energetically close-lying bicyclic 2A2 cation state exhibits low Franck-Condon factors, due to the large change in geometry, and thus cannot be observed.

Studying bicyclists' perceived level of safety using a bicycle simulator combined with immersive virtual reality.

There is a need for methods that provide a better understanding of bicyclists' perceived safety and preferences on currently unavailable and/or unknown bicycle facilities. Different survey methods have been used to study bicyclists' behavior, experiences, and preferences; ranging from verbally described facilities to surveys including images and videos. Virtual Reality (VR) experiments blur the boundaries between stated preference (SP) surveys and revealed preference (RP) surveys and provide a realistic sense of design. This research introduces a novel research method in bicycling research and discusses the results of an experiment using a bicycle simulator combined with immersive VR. In total, 150 participants participated in this experiment and were asked about demographics and perceptions and preferences after bicycling in five different environments with an instrumented bicycle in VR. A 5 × 2 mixed design was used with bicycling environment as within-subject factor and pedestrian / traffic volume as between-subject factor. ANOVA tests revealed how each environment and ambient pedestrian / traffic volume affected perceived level of safety (PLOS) and willingness to bicycle (WTB). Pairwise comparison showed that participants felt safer bicycling on the segregated bicycle path compared to bicycling on the painted bicycle path on the road and roadside. There was no meaningful difference between WTB for less than 10 min and WTB for more than 10 min between bicycling on a painted bicycle path on the sidewalk and painted bicycle path on the road. PLOS and WTB ratings of men and women were not significantly different from each other. The older segment of the sample was more worried about roadside bicycling and bicycle commuters were more confident to ride on the roadside. Despite having several limitations, immersive 360-degree VR was found a powerful presentation tool to evaluate future street designs which can inform transport and urban planning.

Bootstrap parametric GB2 and bootstrap nonparametric distributions for studying shiga toxin-producing Escherichia coli strains growth rate variability.

Shiga toxin-producing Escherichia coli (VTEC) strains, including the serotype O157:H7, are considered foodborne pathogens. Transmission occurs through consumption of undercooked meat, unpasteurized dairy products, vegetables, or contaminated water. The variability of pathogenic and nonpathogenic E. coli strains growth parameters at different temperatures and in different media was studied. Bootstrap parametric (Generalized Beta of the Second Kind, GB2) or nonparametric models were used. GB2 estimations show increased growth rates and shortened lag times with increased temperature, as expected. Similar estimations were obtained using the nonparametric model. Parametric and nonparametric growth rate confidence intervals were wider with increased temperature; lag times confidence intervals were wider with decreased temperature. The nonparametric method gives similar confidence intervals to the parametric method, confirming its suitability for growth parameters estimation. The estimations obtained from nonpathogenic E. coli strains approximate distributions from pathogenic E. coli strains.

Bayesian modeling of two- and three-species bacterial competition in milk.

Listeria monocytogenes is a well-known food-borne pathogen and is among the bacteria best adapted to grow at low temperatures. Psychrotrophic spoilage microorganisms present in milk and milk products are primarily in the genus Pseudomonas, and their numbers increase during cold storage leading to deterioration and/or spoilage. The nature of the competition in two- or three-species bacterial systems with L. monocytogenes, L. innocua, and P. fluorescens in skimmed milk at 7 or 14°C was studied. The Baranyi growth model was used to estimate the growth rate and the maximum population density of the three microorganisms for each strain in single cultures or in two- or three-strains co-cultures. The highest Listeria populations were achieved by pure cultures, decreasing in co-culture with P. fluorescens at both temperatures. A modified deterministic logistic model was applied which includes inhibition functions for single cultures, and two- or three-species cultures. A subsequent Bayesian approach was applied for modelling the bacterial interactions. There was not a direct correlation between the growth rate of P. fluorescens and its inhibitory effect on Listeria species. The use of some species from the natural food microflora to inhibit pathogen growth may be an important tool to enhance the safety of refrigerated foods such as milk and dairy products.

Measuring the equations of state in a relaxed magnetohydrodynamic plasma.

We report measurements of the equations of state of a fully relaxed magnetohydrodynamic (MHD) laboratory plasma. Parcels of magnetized plasma, called Taylor states, are formed in a coaxial magnetized plasma gun, and are allowed to relax and drift into a closed flux conserving volume. Density, ion temperature, and magnetic field are measured as a function of time as the Taylor states compress and heat. The theoretically predicted MHD and double adiabatic equations of state are compared to experimental measurements. We find that the MHD equation of state is inconsistent with our data.

Permutation entropy and statistical complexity analysis of turbulence in laboratory plasmas and the solar wind.

The Bandt-Pompe permutation entropy and the Jensen-Shannon statistical complexity are used to analyze fluctuating time series of three different turbulent plasmas: the magnetohydrodynamic (MHD) turbulence in the plasma wind tunnel of the Swarthmore Spheromak Experiment (SSX), drift-wave turbulence of ion saturation current fluctuations in the edge of the Large Plasma Device (LAPD), and fully developed turbulent magnetic fluctuations of the solar wind taken from the Wind spacecraft. The entropy and complexity values are presented as coordinates on the CH plane for comparison among the different plasma environments and other fluctuation models. The solar wind is found to have the highest permutation entropy and lowest statistical complexity of the three data sets analyzed. Both laboratory data sets have larger values of statistical complexity, suggesting that these systems have fewer degrees of freedom in their fluctuations, with SSX magnetic fluctuations having slightly less complexity than the LAPD edge I(sat). The CH plane coordinates are compared to the shape and distribution of a spectral decomposition of the wave forms. These results suggest that fully developed turbulence (solar wind) occupies the lower-right region of the CH plane, and that other plasma systems considered to be turbulent have less permutation entropy and more statistical complexity. This paper presents use of this statistical analysis tool on solar wind plasma, as well as on an MHD turbulent experimental plasma.

Assessing the Performance of Clostridium perfringens Cooling Models for Cooked, Uncured Meat and Poultry Products.

Heat-resistant spores of Clostridium perfringens may germinate and multiply in cooked meat and poultry products when the rate and extent of cooling does not occur in a timely manner. Therefore, six cooling models (PMP 7.0 broth model; PMIP uncured beef, chicken, and pork models; Smith-Schaffner version 3; and UK IFR ComBase Perfringens Predictor) were evaluated for relative performance in predicting growth of C. perfringens under dynamic temperature conditions encountered during cooling of cooked, uncured meat and poultry products. The predicted growth responses from the models were extensively compared with those observed in food. Data from 188 time-temperature cooling profiles (176 for single-rate exponential cooling and 12 for dual-rate exponential cooling) were collected from 17 independent sources (16 peer-reviewed publications and one report) for model evaluation. Data were obtained for a variety of cooked products, including meat and poultry slurries, ground meat and poultry products with and without added ingredients (e.g., potato starch, sodium triphosphate, and potassium tetrapyrophosphate), and processed products such as ham and roast beef. Performance of the models was evaluated using three sets of criteria, and accuracy was defined within a 1- to 2-log range. The percentages of accurate, fail-safe, or fail-dangerous predictions for each cooling model differed depending on which criterion was used to evaluate the data set. Nevertheless, the combined percentages of accurate and fail-safe predictions based on the three performance criteria were 34.66 to 42.61% for the PMP 7.0 beef broth model, 100% for the PMIP cooling models for uncured beef, uncured pork and uncured chicken, 80.11 to 93.18% for the Smith-Schaffner cooling model, and 74.43 to 85.23% for the UK IFR ComBase Perfringens Predictor model during single-rate exponential chilling. Except for the PMP 7.0 broth model, the other five cooling models (PMIP, Smith-Schaffner, and UK IFR ComBase) are useful and reliable tools that food processors and regulatory agencies can use to evaluate the safety of cooked or heat-treated uncured meat and poultry products exposed to cooling deviations or to develop customized cooling schedules.

Postsplenectomy thrombocytosis: its association with mesenteric, portal, and/or renal vein thrombosis in patients with myeloproliferative disorders.

During the past two years, postoperative thromboses of parts or all of the splanchnic venous circulation have developed in four of 30 patients with myeloproliferative disorders who have undergone splenectomy at our institution. The patients' courses were marked by platelet abnormalities, both quantitative and qualitative. In an attempt to avoid this complication, aspirin and low-dose heparin sodium have been administered and there has been no evidence of thrombosis in the four patients so managed.

The application of the WLF equation to predict lag time as a function of temperature for three psychrotrophic bacteria.

Many mathematical models for microbial growth rates or lag times have been proposed. Most of these models either predict well, but are empirical and offer no insight into mechanism or they reflect mechanism but are too complex or do not fit the data adequately. The Williams-Landel-Ferry (WLF) equation (used by polymer chemists) is empirical, but shows some relation to first principles, and hence may offer some insight into mechanism. This model also has only three parameters and therefore a fairly simple form. The WLF equation was fit to lag times derived from three datasets previously developed in our laboratory. The fits obtained with the WLF equation were as good as the best fits obtained with other models (e.g Arrhenius, Davey, response surface and square root). The WLF model was able to account for 98 to 99% of the variance in the three datasets, indicating a very good fit overall. The parameter estimates of the WLF model were not as highly correlated as those of some of the other models. Many of the models, including the WLF equation did not predict well at very long lag times. Weighted least squares non-linear regression improved the fit for these long lag times.

Application of a statistical bootstrapping technique to calculate growth rate variance for modelling psychrotrophic pathogen growth.

The inherent variability or 'variance' of growth rate measurements is critical to the development of accurate predictive models in food microbiology. A large number of measurements are typically needed to estimate variance. To make these measurements requires a significant investment of time and effort. If a single growth rate determination is based on a series of independent measurements, then a statistical bootstrapping technique can be used to simulate multiple growth rate measurements from a single set of experiments. Growth rate variances were calculated for three large datasets (Listeria monocytogenes, Listeria innocua, and Yersinia enterocolitica) from our laboratory using this technique. This analysis revealed that the population of growth rate measurements at any given condition are not normally distributed, but instead follow a distribution that is between normal and Poisson. The relationship between growth rate and temperature was modeled by response surface models using generalized linear regression. It was found that the assumed distribution (i.e. normal, Poisson, gamma or inverse normal) of the growth rates influenced the prediction of each of the models used. This research demonstrates the importance of variance and assumptions about the statistical distribution of growth rates on the results of predictive microbiological models.

Time-to-detection, percent-growth-positive and maximum growth rate models for Clostridium botulinum 56A at multiple temperatures.

We previously developed models for the influence of inoculum size on the growth kinetics (time-to-detection and maximum growth rate) and percent-growth-positive samples of Clostridium botulinum 56A with factors of inoculum size (1, 100, and 10,000 spores/sample). pH (5.5. 6.0 and 6.5) and sodium chloride concentration (0.5%, 2% and 4%) at 30 degrees C. In this present study, data were collected at two more temperatures (15 and 22 degrees C), making the final design a complete 3 X 3 X 3 X 3 factorial with a total of 81 conditions. Growth was followed hourly as change in A620. The Gompertz equation was fit to the growth data, and the parameters derived were used to calculate the maximum growth rate and time-to-detection. Linear regression with polynomial terms was used to analyze the effect of environmental factors on time-to-detection and maximum growth rate. Logistic regression with polynomial terms was used to analyze the data for percent-growth-positive. Despite the fact that the variance is larger in this extended data set (which includes two temperatures that are further away from the optimum), the inoculum size effect is clearly demonstrated. When inoculum size increased, the percent-growth-positive samples increased and the time-to-detection decreased. When the inoculum was 1000 spores/sample or higher, little additional effect on time-to-detection was observed. Inoculum size might influence results through simple probability or quorum sensing. Our results show that the observed effect of inoculum size from the previous report at a single temperature is not restricted to a specific growth condition, but rather a general phenomenon. The maximum growth rate was independent of inoculum levels, confirming our previous results.

Unanticipated diagnoses found at autopsy in an urban public teaching hospital.

The authors set out to evaluate the use of the autopsy in an urban public teaching hospital setting during the AIDS era. Demographic and length of hospital stay data were obtained from weekly mortality review reports on all patients dying on the medicine service between 1/1/92 and 12/31/93. Clinical and autopsy diagnoses were compared for those patients who had autopsies. The autopsy rate was 16% (152/974). Significant, unsuspected diagnoses were found in 35% (53/152) of the cases, with infections, pulmonary emboli, and myocardial infarctions being most common. Human immunodeficiency virus-infected patients had a greater percentage of unsuspected findings (55%, 23/42), and many of these also were from an infectious etiology. The authors conclude that valuable, unsuspected information frequently can be obtained from autopsies in this clinical setting.

Modeling the survival of Escherichia coli O157:H7 in apple cider using probability distribution functions for quantitative risk assessment.

Outbreaks of foodborne illness from apple cider have prompted research on the survival of Escherichia coli O157:H7 in this food. Published results vary widely, potentially due to differences in E. coli O157:H7 strains, enumeration media, and other experimental considerations. We developed probability distribution functions for the change in concentration of E. coli O157:H7 (log CFU/day) in cider using data from scientific publications for use in a quantitative risk assessment. Six storage conditions (refrigeration [4 to 5 degrees C]; temperature abuse [6 to 10 degrees C]; room temperature [20 to 25 degrees C]; refrigerated with 0.1% sodium benzoate, 0.1% potassium sorbate, or both) were modeled. E. coli survival rate data for all three unpreserved cider storage conditions were highly peaked, and these data were fit to logistic distributions: ideal refrigeration, logistic (-0.061, 0.13); temperature abuse, logistic (-0.0982, 0.23); room temperature, logistic (-0.1, 0.29) and uniform (-4.3, -1.8), to model the very small chance of extremely high log CFU reductions. There were fewer published studies on refrigerated, preserved cider, and these smaller data sets were modeled with beta (4.27, 2.37) x 2.2 - 1.6, normal (-0.2, 0.13), and gamma (1.45, 0.6) distributions, respectively. Simulations were run to show the effect of storage on E. coli O157:H7 during the shelf life of apple cider. Under every storage condition, with and without preservatives, there was an overall decline in E. coli O157:H7 populations in cider, although a small fraction of the time a slight increase was seen.

Glove barriers to bacterial cross-contamination between hands to food.

Human hands are an important source of microbial contamination of foods. However, published data on the effectiveness of handwashing and glove use in a foodservice setting are limited. Bacterial transfer through foodservice quality gloves was quantified using nalidixic acid-resistant Enterobacter aerogenes (a nonpathogenic surrogate with attachment characteristics similar to Salmonella). Five transfer rates were determined: chicken to bare hand, chicken to hand through gloves, bare hand to lettuce, hand to lettuce through gloves (with low inoculum on hands), and hand to lettuce through gloves (with high inoculum on hands). At least 30 observations were made for each percent transfer rate using 30 individual volunteers. The logarithm of percent transfer data were then fit to distributions: chicken to bare hand, normal (0.71, 0.42); chicken to hand through gloves, gamma (5.91, 0.40, -5.00); bare hand to lettuce, logistic (1.16, 0.30); hand to lettuce through gloves (low inoculum), normal (0.35, 0.88); hand to lettuce through gloves (high inoculum), normal (-2.52, 0.61). A 0.01% transfer was observed from food to hands and from hands to food when subjects wore gloves and a 10% transfer was observed without a glove barrier. These results indicate that gloves are permeable to bacteria although transfer from hands to food through a glove barrier was less than without a glove barrier. Our results indicate that gloves may reduce both bacterial transfer from food to the hands of foodservice workers and in subsequent transfer from hands back to food.