Multistate Outbreaks of Foodborne Illness in the United States associated With Fresh Produce From 2010 to 2017

In the United States, the consumption of fresh fruits and vegetables has increased during recent years as consumers seek to make healthier lifestyle choices. However, the number of outbreaks associated with fresh produce that involve cases in more than one state (multistate) has increased concomitantly. As the distance along the farm-to-fork continuum has lengthened over time, there are also more opportunities for fresh produce contamination with bacterial pathogens before it reaches the consumer. This review provides an overview of the three bacterial pathogens (i.e., pathogeni

Food Safety attitudes, Behaviors, and Hygiene Measures among Predominantly Low-income Parents in Houston, Texas

ABSTRACT: Foodborne infections in the United States affect racial-ethnic minority and low-income populations at higher rates than the general population. to identify the prevalence of food safety behaviors and demographic characteristics associated with food handling practices among a susceptible, high-risk population, a cross-sectional survey was administered to 106 parents with children enrolled at two elementary schools serving predominantly low-income families in Houston, Texas. Relationships between demographic characteristics and food safety behavioral outcomes were examined using cross-tabulations and Fisher\u27s exact test. Most respondents were female (93.4%), Hispanic, Latino, or Mexican American (94.9%), and had no previous food handling employment experience (75.0%). The primary source of food safety information reported was the Internet (32.7%), and nearly half of parents (42.7%) reported that they did not consider contamination of food with germs a serious food safety problem. Hand washing before food preparation was more common (98.0%) than before touching the refrigerator handle (66.3%), after electronic device use (55.6%), or after handling raw animal proteins (77.6%). The prevalence of fresh fruit (98.0%) and vegetable (97.9%) washing and appropriate contaminated cutting board handling (89.0%) was high among parents. Self-reported gaps in food handling behaviors identified included lack of food thermometer ownership (80.4%), use of reusable cleaning tools (71.0%), inappropriate defrosting methods (67.4%), and washing of raw poultry (86.3%), seafood (84.9%), and meat (74.7%). Hand washing after electronic device use and defrosting methods were observed to vary significantly according to demographic characteristics. Food safety education with messages targeted to specific demographic groups may be necessary to reduce the risk of foodborne disease among low-income parents and young children

Evaluation of the Kitchen Microbiome and Food Safety Behaviors of Predominantly Low-income Families

Bacterial pathogens in the domestic environment present a risk to residents, particularly among susceptible populations. However, the impact of consumer demographic characteristics and food handling methods on kitchen microbiomes is not fully understood. The domestic kitchen bacterial communities of ten predominantly low-income families in Houston, TX, were assessed in conjunction with a cross-sectional food safety survey to evaluate differences in household and surface-specific microbiomes and bacterial foodborne pathogen presence. Three kitchen surfaces within each household, including the sink drain, the refrigerator handle, and the counter, were environmentally sampled and metataxonomically evaluated via targeted 16S rRNA sequencing. Disposable dish sponges were also acquired and examined. Results indicated that alpha diversity did not vary by the households, sampling locations, or demographic characteristics evaluated. Significant differences in beta diversity were observed among the bacterial communities of five pairs of households and between refrigerator handle and disposable dish sponge microbiomes. A total of 89 unique bacterial foodborne pathogens were identified across surface types. Each household contained at least one contaminated surface, and the most common bacterial foodborne pathogens identified wer

THE TRANSCRIPTIONAL RESPONSE OF ARTIFICIALLY-INOCULATED LISTERIA MONOCYTOGENES DURING THE MANUFACTURE OF UNPASTEURIZED GOUDA CHEESE

Listeriosis outbreaks indicate that L. monocytogenes contamination is an issue for various food types, such as unpasteurized (raw) cheese. Current regulation prevents the interstate sale and distribution of raw cheese and mandates a ≥60 day aging period at ≥2˚C to ensure product safety; yet studies demonstrate that pathogens can persist during aging. Environmental stress can alter the transcriptomic profiles of pathogens; however, these surveys are rarely conducted in food matrices. This study aimed to assess the transcriptomic profiles of L. monocytogenes strain F2365 during environmental stressors inherent throughout cheesemaking. First, quantitative polymerase chain reaction (qPCR) was used to monitor transcription levels of nine L. monocytogenes genes involved in virulence, stress response, energy transport, and metabolism during osmotic stress (11% NaCl) and varying temperature/time (5˚C 24 h, 25˚C 24 h, 38˚C 30 min) conditions in Brain Heart Infusion (BHI) broth, raw milk, and pasteurized milk. Generally, the genes prfA, lmo1381, lmo0963, and lmo1875 were down-regulated, whereas the genes lmo1864, lmo0914, lmo0348, lmo1428, and lmo1264 were up-regulated. Virulence gene, prfA, was most down-regulated when L. monocytogenes was grown in raw milk with salt at 25˚C (4774.72±838.14 fold; relative to 24 h growth at 37˚C). The stress response gene lmo0914, encoding σB, was most up-regulated when L. monocytogenes was grown in BHI at 25˚C with salt (14.61±7.72 fold). Additionally, transcription levels of the nine genes were assessed at points during the laboratory-scale manufacture of Gouda cheese made with raw milk artificially-inoculated with L. monocytogenes via qPCR. Similar differential regulation for both prfA and lmo0914 in L. monocytogenes was observed during cheesemaking. The gene lmo1864, encoding a putative pore-forming hemolysin, was up-regulated throughout the cheesemaking process, but was most up-regulated after stirring the curd (449.81±432.53 fold). Ultimately, these results indicate that the lmo1864 gene may play a role in L. monocytogenes survival during cheesemaking. Methods developed in this study can be used to assess the risk of L. monocytogenes, not only during cheesemaking, but during the ≥60-day aging process. Overall, these results contribute to the understanding of L. monocytogenes survival mechanisms during the cheesemaking process.M.S. in Food Safety and Technology, May 201

Data_Sheet_1_Evaluation of the kitchen microbiome and food safety behaviors of predominantly low-income families.docx

Bacterial pathogens in the domestic environment present a risk to residents, particularly among susceptible populations. However, the impact of consumer demographic characteristics and food handling methods on kitchen microbiomes is not fully understood. The domestic kitchen bacterial communities of ten predominantly low-income families in Houston, TX, were assessed in conjunction with a cross-sectional food safety survey to evaluate differences in household and surface-specific microbiomes and bacterial foodborne pathogen presence. Three kitchen surfaces within each household, including the sink drain, the refrigerator handle, and the counter, were environmentally sampled and metataxonomically evaluated via targeted 16S rRNA sequencing. Disposable dish sponges were also acquired and examined. Results indicated that alpha diversity did not vary by the households, sampling locations, or demographic characteristics evaluated. Significant differences in beta diversity were observed among the bacterial communities of five pairs of households and between refrigerator handle and disposable dish sponge microbiomes. A total of 89 unique bacterial foodborne pathogens were identified across surface types. Each household contained at least one contaminated surface, and the most common bacterial foodborne pathogens identified were Escherichia coli, Staphylococcus aureus, and Klebsiella pneumoniae. All parents reported washing their hands before meal preparation, washing fresh fruits and vegetables, and washing cutting boards with soap after use to prepare raw animal proteins. Gaps in food safety behaviors identified included a lack of serious concern for food contamination with germs and inappropriate handwashing, food handling, and cleaning behaviors. The number of unique bacterial foodborne pathogens identified within households was significantly higher among households whose respondent parent reported that they did not consider food contamination with germs to be a serious food safety problem (median: 41.0 species) compared to households whose respondent parent did consider food contamination to be a serious food safety problem (median: 3.0 species; p value = 0.0218). These results demonstrate that domestic kitchen taxonomic abundance profiles vary according to household and surface type. Data suggest that low-income consumers may be at risk of foodborne pathogen exposure from contaminated home kitchen surfaces, and that food safety attitudes may directly contribute to this hazard.</p

Listeria monocytogenes Growth Kinetics in Milkshakes Made from Naturally and Artificially Contaminated Ice Cream

This study assessed the growth of Listeria monocytogenes in milkshakes made using the process-contaminated ice cream associated with a listeriosis outbreak in comparison to milkshakes made with artificially contaminated ice cream. For all temperatures, growth kinetics including growth rates, lag phases, maximum populations, and population increases were determined for the naturally and artificially derived contaminants at 5, 10, 15, and 25°C storage for 144 h. The artificially inoculated L. monocytogenes presented lower growth rates and shorter lag phases than the naturally contaminated populations at all temperatures except for 5°C, where the reverse was observed. At 25°C, lag phases of the naturally and artificially contaminated L. monocytogenes were 11.6 and 7.8 h, respectively. The highest increase in population was observed for the artificially inoculated pathogen at 15°C after 96 h (6.16 log CFU/mL) of storage. Growth models for both contamination states in milkshakes were determined. In addition, this study evaluated the antimicrobial effectiveness of flavoring agents, including strawberry, chocolate and mint, on the growth of the pathogen in milkshakes during 10°C storage. All flavor additions resulted in decreased growth rates of L. monocytogenes for both contamination states. The addition of chocolate and mint flavoring also resulted in significantly longer lag phases for both contamination states. This study provides insight into the differences in growth between naturally and artificially contaminated L. monocytogenes in a food product

Metagenomics of pasteurized and unpasteurized gouda cheese using targeted 16S rDNA sequencing

Abstract Background The microbiome of cheese is diverse, even within a variety. The metagenomics of cheese is dependent on a vast array of biotic and abiotic factors. Biotic factors include the population of microbiota and their resulting cellular metabolism. Abiotic factors, including the pH, water activity, fat, salt, and moisture content of the cheese matrix, as well as environmental conditions (temperature, humidity, and location of aging), influence the biotic factors. This study assessed the metagenomics of commercial Gouda cheese prepared using pasteurized or unpasteurized cow milk or pasteurized goat milk via 16S rDNA sequencing. Results Results were analyzed and compared based on milk pasteurization and source, spatial variability (core, outer, and under the rind), and length of aging (2–4 up to 12–18 months). The dominant organisms in the Gouda cheeses, based on percentage of sequence reads identified at the family or genus levels, were Bacillaceae, Lactococcus, Lactobacillus, Streptococcus, and Staphylococcus. More genus- or family-level (e.g. Bacillaceae) identifications were observed in the Gouda cheeses prepared with unpasteurized cow milk (120) compared with those prepared with pasteurized cow milk (92). When assessing influence of spatial variability on the metagenomics of the cheese, more pronounced differences in bacterial genera were observed in the samples taken under the rind; Brachybacterium, Pseudoalteromonas, Yersinia, Klebsiella, and Weissella were only detected in these samples. Lastly, the aging length of the cheese greatly influenced the number of organisms observed. Twenty-seven additional genus-level identifications were observed in Gouda cheese aged for 12–18 months compared with cheese only aged 2–4 months. Conclusions Collectively, the results of this study are important in determining the typical microbiota associated with Gouda cheese and how the microbiome plays a role in safety and quality

Systemic Inflammation in Preclinical Ulcerative Colitis

Background &amp; Aims: Preclinical ulcerative colitis is poorly defined. We aimed to characterize the preclinical systemic inflammation in ulcerative colitis, using a comprehensive set of proteins. Methods: We obtained plasma samples biobanked from individuals who developed ulcerative colitis later in life (n = 72) and matched healthy controls (n = 140) within a population-based screening cohort. We measured 92 proteins related to inflammation using a proximity extension assay. The biologic relevance of these findings was validated in an inception cohort of patients with ulcerative colitis (n = 101) and healthy controls (n = 50). To examine the influence of genetic and environmental factors on these markers, a cohort of healthy twin siblings of patients with ulcerative colitis (n = 41) and matched healthy controls (n = 37) were explored. Results: Six proteins (MMP10, CXCL9, CCL11, SLAMF1, CXCL11 and MCP-1) were up-regulated (P &lt; .05) in preclinical ulcerative colitis compared with controls based on both univariate and multivariable models. Ingenuity Pathway Analyses identified several potential key regulators, including interleukin-1β, tumor necrosis factor, interferon-gamma, oncostatin M, nuclear factor-κB, interleukin-6, and interleukin-4. For validation, we built a multivariable model to predict disease in the inception cohort. The model discriminated treatment-naïve patients with ulcerative colitis from controls with leave-one-out cross-validation (area under the curve = 0.92). Consistently, MMP10, CXCL9, CXCL11, and MCP-1, but not CCL11 and SLAMF1, were significantly up-regulated among the healthy twin siblings, even though their relative abundances seemed higher in incident ulcerative colitis. Conclusions: A set of inflammatory proteins are up-regulated several years before a diagnosis of ulcerative colitis. These proteins were highly predictive of an ulcerative colitis diagnosis, and some seemed to be up-regulated already at exposure to genetic and environmental risk factors