Quantitative risk assessment model to investigate the public health impact of varying Listeria monocytogenes allowable levels in different food commodities: A retrospective analysis.

Invasive listeriosis is a potentially fatal foodborne disease that according to this study may affect up to 32.9 % of the US population considered as increased risk and including people with underlying conditions and co-morbidities. Listeria monocytogenes has been scrutinized in research and surveillance programs worldwide in Ready-to-Eat (RTE) food commodities (RTE salads, deli meats, soft/semi-soft cheese, seafood) and frozen vegetables in the last 30 years with an estimated overall prevalence of 1.4-9.9 % worldwide (WD) and 0.5-3.8 % in the United States (US). Current L. monocytogenes control efforts have led to a prevalence reduction in the last 5 years of 4.9-62.9 % (WD) and 12.4-92.7 % (US). A quantitative risk assessment model was developed, estimating the probability of infection in the US susceptible population to be 10-10,000× higher than general population and the total number of estimated cases in the US was 1044 and 2089 cases by using the FAO/WHO and Pouillot dose-response models. Most cases were attributed to deli meats (>90 % of cases) followed by RTE salads (3.9-4.5 %), soft and semi-soft cheese and RTE seafood (0.5-1.0 %) and frozen vegetables (0.2-0.3 %). Cases attributed to the increased risk population corresponded to 96.6-98.0 % of the total cases with the highly susceptible population responsible for 46.9-80.1 % of the cases. Removing product lots with a concentration higher than 1 CFU/g reduced the prevalence of contamination by 15.7-88.3 % and number of cases by 55.9-100 %. Introducing lot-by-lot testing and defining allowable quantitative regulatory limits for low-risk RTE commodities may reduce the public health impact of L. monocytogenes and improve the availability of enumeration data.

The power, potential, benefits, and challenges of implementing high-throughput sequencing in food safety systems.

The development and application of modern sequencing technologies have led to many new improvements in food safety and public health. With unprecedented resolution and big data, high-throughput sequencing (HTS) has enabled food safety specialists to sequence marker genes, whole genomes, and transcriptomes of microorganisms almost in real-time. These data reveal not only the identity of a pathogen or an organism of interest in the food supply but its virulence potential and functional characteristics. HTS of amplicons, allow better characterization of the microbial communities associated with food and the environment. New and powerful bioinformatics tools, algorithms, and machine learning allow for development of new models to predict and tackle important events such as foodborne disease outbreaks. Despite its potential, the integration of HTS into current food safety systems is far from complete. Government agencies have embraced this new technology, and use it for disease diagnostics, food safety inspections, and outbreak investigations. However, adoption and application of HTS by the food industry have been comparatively slow, sporadic, and fragmented. Incorporation of HTS by food manufacturers in their food safety programs could reinforce the design and verification of effectiveness of control measures by providing greater insight into the characteristics, origin, relatedness, and evolution of microorganisms in our foods and environment. Here, we discuss this new technology, its power, and potential. A brief history of implementation by public health agencies is presented, as are the benefits and challenges for the food industry, and its future in the context of food safety.

Prevalence of Listeria Species and Listeria monocytogenes on Raw Produce Arriving at Frozen Food Manufacturing Facilities.

ABSTRACT: The ubiquity of Listeria monocytogenes in the environment affects the food industry and presents concerns for frozen food facilities. This study determined the prevalence and numbers of Listeria species and L. monocytogenes on raw produce arriving at frozen food facilities. Raw produce was collected using multilevel blinding protocols to ensure anonymity of participants and avoid traceback. Five raw vegetables were selected: corn, carrots, green beans, peas, and spinach. Raw products were collected after arrival at the facilities but before cleaning or other preprocessing steps that are typically performed inside the facility. The U.S. Food and Drug Administration's Bacteriological Analytical Manual method for detection of Listeria spp. and L. monocytogenes was followed, with PCR screening followed by selective plating methods. Listeria numbers were estimated from positive samples using the most-probable-number (MPN) methodology. A total of 290 samples were collected, with 96 and 17 samples positive for Listeria spp. (33.1%) and L. monocytogenes (5.9%), respectively. Enumeration data for the 96 Listeria spp. samples indicated 82 samples had greater than 100 MPN of Listeria spp. per g and 14 samples had less than 100 MPN Listeria spp. per g. The prevalence of Listeria spp. varied by commodity: spinach (66.7%), peas (50%), corn (32.2%), green beans (22.2%), and carrots (13%). L. monocytogenes prevalence was determined in corn (13.6%), peas (6.3%), and green beans (4.2%) arriving at processing facilities. Such data were previously unavailable to frozen vegetable processors and are valuable in implementing process control standards. The prevalence and pathogen concentration data from raw commodities found in this study can provide the industry with information to conduct more accurate quantitative risk assessments and a baseline to model and target appropriate pathogen reduction steps during processing.

Blinding Protocols for Acquisition of Potentially Sensitive Food Safety Information.

ABSTRACT: Difficulties associated with addressing research problems can revolve around the collection of data from private entities. Potential issues can arise when collecting food samples or food safety data from industry or third-party sources because of concerns about distribution or exposure of potentially sensitive information. Industry is cautious of its involvement in research projects because issues associated with production levels, capital investment, regulatory inquiries, unwarranted publicity, or other legal aspects can arise depending on the nature of information gathered, and information may be inadvertently released into the public domain. Well-designed clinical trials with animals or humans use blinding methods to reduce bias. In this study, a similar strategy was applied to acquisition of sensitive data to gather meaningful food safety related data while assuring that information provided was not at risk. Blinding methods for collecting electronic data and material samples were created to obtain materials and records directly from participating frozen food companies. This approach provided insight into current industry practices without potential downsides for participating companies. Analysis of food safety concerns using industry data and the distribution of findings can be of assistance industry-wide for conducting risk assessments and developing improved research-based food safety plans. The method described was designed to collect information using blinding protocols to reduce bias and prevent traceback to the original source. The use of blinding protocols promotes industry participation and creates data collection with anonymity for the original source, which can improve reliability of the research and applicability for industry. These blinding protocols are suitable for use in future food safety research projects involving data within and between various segments of the food industry and could be used to encourage collection of valuable industry samples and data.

Evaluating Environmental Monitoring Protocols for Listeria spp. and Listeria monocytogenes in Frozen Food Manufacturing Facilities.

Food processors face serious challenges due to Listeria monocytogenes contamination. Environmental monitoring is used to control L. monocytogenes from the processing environment. Although frozen foods do not support the growth of L. monocytogenes, the moist and cold conditions in frozen food production environments are favorable for growth of L. monocytogenes. The purpose of the study was to determine the current state of awareness and practices applied across a variety of frozen food facilities related to environmental monitoring for Listeria. A survey tool was created to elicit information on existing environmental monitoring programs within the frozen food industry. The topics included cleaning and sanitizing applications and frequency, microbiological testing, and environmental areas of concern. The survey was reviewed by academic and industry experts with knowledge of microbiology and frozen food processing and was field tested by industry personnel with extensive knowledge of environmental monitoring. The survey was distributed and analyzed electronically via Qualtrics among 150 frozen food contacts. Data were gathered anonymously with a response rate of 31% (n = 46). The survey indicated that facilities are more likely to test for Listeria spp. in environmental monitoring zones 2 to 4 (nonfood contact areas) on a weekly basis. The major areas of concern in facilities for finding Listeria-positive results are floors, walls, and drains. At the time of the survey, few facilities incorporated active raw material and finished product testing for Listeria; instead, programs emphasized the need to identify presence of Listeria in the processing environment and mitigate potential for product contamination. Recognition of environmental monitoring as a key component of a comprehensive food safety plan was evident, along with an industry focus to further improve and develop verification programs to reduce prevalence of L. monocytogenes in frozen food processing environments.

Advances in the understanding of dairy and cheese flavors: symposium introduction.

A symposium titled "Advances in the Understanding of Dairy and Cheese Flavors" was held in September 2013 at the American Chemical Society's 246th National Meeting in Indianapolis, IN, USA. The symposium, which was sponsored by the Division of Agricultural and Food Chemistry, was to discuss the state of the art in the detection and quantitation of flavor in dairy products. The authors of two of the presentations have been selected to expand on their talks by submitting full papers about their research.

Virus assay using antibody-functionalized peptide nanotubes.

Robust trace-level detection of viruses is crucial to meet urgent needs in fighting the spread of disease or detecting bioterrorism events. We report a new method for rapid and highly sensitive detection of viruses utilizing fluorescent antibody nanotubes. When viral pathogens were mixed with these antibody nanotubes, the nanotubes rapidly aggregated around the viruses to form a networking structure. Trace quantities of viruses such as herpes simplex virus type 2, adenovirus, vaccinia and influenza type B were detected on attomolar order by changes in fluorescence and light scattering intensities associated with aggregation of dye-loaded antibody nanotubes around viruses. High specificity of each antibody nanotube toward its targeted virus was demonstrated by quantifying concentrations of two different viruses in mixtures. This antibody nanotube assay detects targeted pathogens within 30 minutes after incubation with antibody nanotubes. This antibody nanotube assay could fill a pressing need to detect and quantify viruses both rapidly and sensitively.

Overexpression of Lactobacillus casei D-hydroxyisocaproic acid dehydrogenase in cheddar cheese.

Metabolism of aromatic amino acids by lactic acid bacteria is an important source of off-flavor compounds in Cheddar cheese. Previous work has shown that alpha-keto acids produced from Trp, Tyr, and Phe by aminotransferase enzymes are chemically labile and may degrade spontaneously into a variety of off-flavor compounds. However, dairy lactobacilli can convert unstable alpha-keto acids to more-stable alpha-hydroxy acids via the action of alpha-keto acid dehydrogenases such as d-hydroxyisocaproic acid dehydrogenase. To further characterize the role of this enzyme in cheese flavor, the Lactobacillus casei d-hydroxyisocaproic acid dehydrogenase gene was cloned into the high-copy-number vector pTRKH2 and transformed into L. casei ATCC 334. Enzyme assays confirmed that alpha-keto acid dehydrogenase activity was significantly higher in pTRKH2:dhic transformants than in wild-type cells. Reduced-fat Cheddar cheeses were made with Lactococcus lactis starter only, starter plus L. casei ATCC 334, and starter plus L. casei ATCC 334 transformed with pTRKH2:dhic. After 3 months of aging, the cheese chemistry and flavor attributes were evaluated instrumentally by gas chromatography-mass spectrometry and by descriptive sensory analysis. The culture system used significantly affected the concentrations of various ketones, aldehydes, alcohols, and esters and one sulfur compound in cheese. Results further indicated that enhanced expression of d-hydroxyisocaproic acid dehydrogenase suppressed spontaneous degradation of alpha-keto acids, but sensory work indicated that this effect retarded cheese flavor development.