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Concept: Foodborne illness

312

In order to facilitate foodborne outbreak investigations there is a need to improve the methods for identifying the food products that should be sampled for laboratory analysis. The aim of this study was to examine the applicability of a likelihood ratio approach previously developed on simulated data, to real outbreak data. We used human case and food product distribution data from the Norwegian enterohaemorrhagic Escherichia coli outbreak in 2006. The approach was adjusted to include time, space smoothing and to handle missing or misclassified information. The performance of the adjusted likelihood ratio approach on the data originating from the HUS outbreak and control data indicates that the adjusted approach is promising and indicates that the adjusted approach could be a useful tool to assist and facilitate the investigation of food borne outbreaks in the future if good traceability are available and implemented in the distribution chain. However, the approach needs to be further validated on other outbreak data and also including other food products than meat products in order to make a more general conclusion of the applicability of the developed approach.

Concepts: Bacteria, Nutrition, Escherichia coli, Food, Escherichia coli O157:H7, Foodborne illness, Food safety, Maslow's hierarchy of needs

169

Microbial communities associated with agricultural animals are important for animal health, food safety, and public health. Here we combine high-throughput sequencing (HTS), quantitative-PCR assays, and network analysis to profile the poultry-associated microbiome and important pathogens at various stages of commercial poultry production from the farm to the consumer. Analysis of longitudinal data following two flocks from the farm through processing showed a core microbiome containing multiple sequence types most closely related to genera known to be pathogenic for animals and/or humans, including , and . After the final stage of commercial poultry processing, taxonomic richness was ca. 2-4 times lower than the richness of fecal samples from the same flocks and abundance was significantly reduced. Interestingly, however, carcasses sampled at 48 hr after processing harboured the greatest proportion of unique taxa (those not encountered in other samples), significantly more than expected by chance. Among these were anaerobes such as , , , and multiple sequence types. Retail products were dominated by , but also contained 27 other genera, most of which were potentially metabolically active and encountered in on-farm samples. Network analysis was focused on the foodborne pathogen and revealed a majority of sequence types with no significant interactions with other taxa, perhaps explaining the limited efficacy of previous attempts at competitive exclusion of . These data represent the first use of HTS to characterize the poultry microbiome across a series of farm-to-fork samples and demonstrate the utility of HTS in monitoring the food supply chain and identifying sources of potential zoonoses and interactions among taxa in complex communities.

Concepts: Bacteria, Microbiology, Food, Pathogen, Sequence, Foodborne illness, Poultry farming, Poultry

167

This work explores the bactericidal effect of (+)-limonene, the major constituent of citrus fruits' essential oils, against E. coli. The degree of E. coli BJ4 inactivation achieved by (+)-limonene was influenced by the pH of the treatment medium, being more bactericidal at pH 4.0 than at pH 7.0. Deletion of rpoS and exposure to a sub-lethal heat or an acid shock did not modify E. coli BJ4 resistance to (+)-limonene. However, exposure to a sub-lethal cold shock decreased its resistance to (+)-limonene. Although no sub-lethal injury was detected in the cell envelopes after exposure to (+)-limonene by the selective-plating technique, the uptake of propidium iodide by inactivated E. coli BJ4 cells pointed out these structures as important targets in the mechanism of action. Attenuated Total Reflectance Infrared Microspectroscopy (ATR-IRMS) allowed identification of altered E. coli BJ4 structures after (+)-limonene treatments as a function of the treatment pH: β-sheet proteins at pH 4.0 and phosphodiester bonds at pH 7.0. The increased sensitivity to (+)-limonene observed at pH 4.0 in an E. coli MC4100 lptD4213 mutant with an increased outer membrane permeability along with the identification of altered β-sheet proteins by ATR-IRMS indicated the importance of this structure in the mechanism of action of (+)-limonene. The study of mechanism of inactivation by (+)-limonene led to the design of a synergistic combined process with heat for the inactivation of the pathogen E. coli O157:H7 in fruit juices. These results show the potential of (+)-limonene in food preservation, either acting alone or in combination with lethal heat treatments.

Concepts: DNA, Bacteria, Escherichia coli, Citrus, Escherichia coli O157:H7, Fruit, Foodborne illness, Juice vesicles

141

An outbreak of salmonellosis occurred following attendance at a school camp between 5 and 8 August 2014 in a remote area of the Northern Territory, Australia. We conducted a retrospective cohort study via telephone interviews, using a structured questionnaire that recorded symptoms and exposures to foods and activities during the camp. A case was anyone with laboratory confirmed Salmonella Saintpaul infection or a clinically compatible illness after attending the camp. Environmental health officers from the Environmental Health Branch undertook an investigation and collected water and environmental samples. We interviewed 65 (97%) of the 67 people who attended the camp. There were 60 students and 7 adults. Of the 65 people interviewed, 30 became ill (attack rate 46%); all were students; and 4 had laboratory confirmed S. Saintpaul infection. The most commonly reported symptoms were diarrhoea (100% 30/30), abdominal pain (93% 28/30), nausea (93% 28/30) and fever (70% 21/30). Thirteen people sought medical attention but none required hospitalisation. Illness was significantly associated with drinking cordial at lunch on 7 August (RR 3.8, 95% CI 1.3-11, P < 0.01), as well as drinking cordial at lunch on 8 August (RR 2.1, 95% CI 1.1-4.2, P=0.01). Salmonella spp. was not detected in water samples or wallaby faeces collected from the camp ground. The epidemiological investigation suggests the outbreak was caused by environmental contamination of food or drink and could have occurred during ice preparation or storage, preparation of the cordial or from inadequate sanitising of the cooler from which the cordial was served. This outbreak highlights the risks of food or drink contamination with environmental Salmonella. Those preparing food and drink in campground settings should be vigilant with cleaning, handwashing and disinfection to prevent outbreaks of foodborne disease.

Concepts: Cohort study, Epidemiology, Illness, Salmonella, Diarrhea, Foodborne illness, Gastroenteritis, Environmental health officer

141

To investigate whether acid suppression medicines (ASMs) increase the risk of bacterial gastroenteritis.

Concepts: Cohort study, Diarrhea, Foodborne illness

122

Bacterial cross-contamination from surfaces to food can contribute to foodborne disease. The cross-contamination rate of Enterobacter aerogenes was evaluated on household surfaces using scenarios that differed by surface type, food type, contact time (<1, 5, 30 and 300 s), and inoculum matrix (tryptic soy broth or peptone buffer). The surfaces used were stainless steel, tile, wood and carpet. The food types were watermelon, bread, bread with butter and gummy candy. Surfaces (25 cm(2)) were spot inoculated with 1 ml of inoculum and allowed to dry for 5 h, yielding an approximate concentration of 10(7) CFU/surface. Foods (with 16 cm(2) contact area) were dropped on the surfaces from a height of 12.5 cm and left to rest as appropriate. Post transfer surfaces and foods were placed in sterile filter bags and homogenized or massaged, diluted and plated on tryptic soy agar. The transfer rate was quantified as the log % transfer from the surface to the food. Contact time, food and surface type all had a highly significant effect (P<0.000001) on log % transfer of bacteria. The inoculum matrix (TSB or peptone buffer) also had a significant effect on transfer (P = 0.013), and most interaction terms were significant. More bacteria transferred to watermelon (∼0.2-97%) relative to other foods, while fewer bacteria transferred to gummy candy (∼0.1-62%). Transfer of bacteria to bread (∼0.02-94%) and bread with butter (∼0.02-82%) were similar, and transfer rates under a given set of condition were more variable compared with watermelon and gummy candy.

Concepts: Bacteria, Food, Bread, Foodborne illness, Food safety, Enterobacter, Enterobacter aerogenes, Gummi bear

111

The U.S. wastes 31 to 40% of its post-harvest food supply, with a substantial portion of this waste occurring at the consumer level. Globally, interventions to address wasted food have proliferated, but efforts are in their infancy in the U.S. To inform these efforts and provide baseline data to track change, we performed a survey of U.S. consumer awareness, attitudes and behaviors related to wasted food. The survey was administered online to members of a nationally representative panel (N=1010), and post-survey weights were applied. The survey found widespread (self-reported) awareness of wasted food as an issue, efforts to reduce it, and knowledge about how to do so, plus moderately frequent performance of waste-reducing behaviors. Three-quarters of respondents said they discard less food than the average American. The leading motivations for waste reduction were saving money and setting an example for children, with environmental concerns ranked last. The most common reasons given for discarding food were concern about foodborne illness and a desire to eat only the freshest food. In some cases there were modest differences based on age, parental status, and income, but no differences were found by race, education, rural/urban residence or other demographic factors. Respondents recommended ways retailers and restaurants could help reduce waste. This is the first nationally representative consumer survey focused on wasted food in the U.S. It provides insight into U.S. consumers' perceptions related to wasted food, and comparisons to existing literature. The findings suggest approaches including recognizing that many consumers perceive themselves as being already-knowledgeable and engaged, framing messages to focus on budgets, and modifying existing messages about food freshness and aesthetics. This research also suggests opportunities to shift retail and restaurant practice, and identifies critical research gaps.

Concepts: Food, Recycling, Foodborne illness, Food safety, Restaurant, Consumer

103

Backyard poultry flocks have increased in popularity concurrent with an increase in live poultry-associated salmonellosis (LPAS) outbreaks. Better understanding of practices that contribute to this emerging public health issue is needed. We reviewed outbreak reports to describe the epidemiology of LPAS outbreaks in the United States, examine changes in trends, and inform prevention campaigns. LPAS outbreaks were defined as ≥2 culture-confirmed human Salmonella infections linked to live poultry contact. Outbreak data were obtained through multiple databases and a literature review. During 1990-2014, a total of 53 LPAS outbreaks were documented, involving 2,630 illnesses, 387 hospitalizations, and 5 deaths. Median patient age was 9 years (range <1 to 92 years). Chick and duckling exposure were reported by 85% and 38% of case-patients, respectively. High-risk practices included keeping poultry inside households (46% of case-patients) and kissing birds (13%). Comprehensive One Health strategies are needed to prevent illnesses associated with live poultry.

Concepts: Health care, Public health, Health, Epidemiology, United States, Salmonella, Foodborne illness, Salmonellosis

67

Identifying the major sources of risk in disease transmission is key to designing effective controls. However, understanding of transmission dynamics across species boundaries is typically poor, making the design and evaluation of controls particularly challenging for zoonotic pathogens. One such global pathogen is Escherichia coli O157, which causes a serious and sometimes fatal gastrointestinal illness. Cattle are the main reservoir for E. coli O157, and vaccines for cattle now exist. However, adoption of vaccines is being delayed by conflicting responsibilities of veterinary and public health agencies, economic drivers, and because clinical trials cannot easily test interventions across species boundaries, lack of information on the public health benefits. Here, we examine transmission risk across the cattle-human species boundary and show three key results. First, supershedding of the pathogen by cattle is associated with the genetic marker stx2. Second, by quantifying the link between shedding density in cattle and human risk, we show that only the relatively rare supershedding events contribute significantly to human risk. Third, we show that this finding has profound consequences for the public health benefits of the cattle vaccine. A naïve evaluation based on efficacy in cattle would suggest a 50% reduction in risk; however, because the vaccine targets the major source of human risk, we predict a reduction in human cases of nearly 85%. By accounting for nonlinearities in transmission across the human-animal interface, we show that adoption of these vaccines by the livestock industry could prevent substantial numbers of human E. coli O157 cases.

Concepts: Epidemiology, Bacteria, Vaccine, Vaccination, Escherichia coli, Escherichia coli O157:H7, Shiga toxin, Foodborne illness

66

Crohn’s disease (CD) is a chronic inflammatory condition of diverse etiology. Exposure to foodborne pathogens causing acute gastroenteritis produces a long-term risk of CD well into the post-infectious period but the mechanistic basis for this ongoing relationship to disease onset is unknown. We developed two novel models to study the comorbidity of acute gastroenteritis caused by Salmonella Typhimurium or Citrobacter rodentium in mice colonized with adherent-invasive Escherichia coli (AIEC), a bacterial pathobiont linked to CD. Here, we show that disease activity in the post-infectious period after gastroenteritis is driven by the tissue-associated expansion of the resident AIEC pathobiont, with an attendant increase in immunopathology, barrier defects, and delays in mucosal restitution following pathogen clearance. These features required AIEC resistance to host defense peptides and a fulminant inflammatory response to the enteric pathogen. Our results suggest that individuals colonized by AIEC at the time of acute infectious gastroenteritis may be at greater risk for CD onset. Importantly, our data identify AIEC as a tractable disease modifier, a finding that could be exploited in the development of therapeutic interventions following infectious gastroenteritis in at-risk individuals.

Concepts: Immune system, Bacteria, Microbiology, Escherichia coli, Salmonella, Diarrhea, Foodborne illness, Gastroenteritis