This chapter discusses the great involvement of home-processed foods in outbreaks of botulism. In general, foodborne botulism is caused by eating improperly preserved food in which Clostridium botulinumhas grown and produced toxin. The etiologic agent of botulism was first isolated in 1896 by Emile Pierre Marie van Ermengem, a professor of bacteriology at the University of Ghent in Belgium. In infant botulism, spores of C. botulinum are ingested; they colonize the intestine, germinate, and form botulinal toxin in the intestinal lumen. Conventional botulism food poisoning, or botulism, results from consumption of food that has supported growth of C. botulinum and elaboration of its deadly neurotoxin. Spores prepared from cultures freshly isolated from a botulism outbreak produced toxin more readily in the implicated food than spores of laboratory stock cultures. Botulism as a disease was first recognized in the United States in 1899, and since then records of reported outbreaks have been maintained.

ObjectivesRendered oils and fats are commonly used in pet food products. Given the implementation of the Food Safety Modernization Act (FSMA), which enforces a zero-tolerance policy for Salmonella in pet foods, the microbiological safety of these products has become an area of focus for the industry. The objectives of this study were to (i) understand the lethality of E. coli O157:H7 and Salmonella in heat treated, rendered used cooking oil, (ii) determine D and z-values for E. coli O157:H7 and Salmonella in heat treated, rendered used cooking oil and (iii) establish critical limits for use in a HACCP or other food safety systems.Materials and MethodsThe objectives were addressed by inoculating previously rendered, used cooking oil, with a five-strain mixture of Salmonella or E. coli O157:H7 at a level of approximately 8.0 log CFU/ml. Inoculated samples were exposed to heat at 62, 71, or 82°C using a water bath fitted with a circulator. At predetermined time intervals, individual samples were removed from water bath and immediately chilled in an ice bath. Samples were diluted in 0.1% buffered peptone water supplemented with 1% emulsifier (Tween 80). Samples were plated on selective agars (xylose lysine deoxycholate [Salmonella]; sorbitol MacConkey [E. coli O157:H7]) and a non-selective agar (tryptic soy agar supplemented with 1% sodium pyruvate). Limited methodologies are available for the microbiological analysis of high fat and/or oil matrices. Extensive pre-project work was conducted to establish appropriate methodologies to meet objectives in a laboratory setting. It was determined that a traditionally used aqueous inoculum would not be adequately distributed throughout an oil matrix and could potentially interfere with heat transfer during heat treatment. Thus, pathogen cells were resuspended in warmed (37°C), sterile vegetable oil (soybean oil). No difference in inoculation level was observed when an aqueous solution and sterile vegetable oil were compared. Volumetric measurement of oil samples proved inconsistent and thus for experimentation, the mass, rather than volume, was utilized. The addition of Tween 80 aided in homogenization of sample and diluent during surface plating and eliminated phase separation and errors in dilution (i.e., no ten-fold differences across dilutions). Additionally, a larger width tube (50 mL Falcon Tube) was used for the first dilution blank and vortexing was standardized to 30 s—these components proved to be critical for microbial analysis of high-fat liquids or semi-solids. Six replicates were performed for each pathogen and temperature combination. Segmented regression was performed using the Proc NLIN function of SAS (v 9.4; SAS Inst. Inc., Cary, NC) and z-values were calculated using linear regression (SAS).ResultsNonlinear death curves were observed for all pathogen and temperature combinations. This response to heat treatment indicated not only pathogen survival, but possible heat resistance among some strains. As expected, D-values were lower as temperature increased and for all pathogens ranged from 0.03 to 0.04 min at 82°C, 0.14 to 0.27 min at 71°C, and 0.77 to 1.49 min at 62°C. Similarly, Z-values ranged from 14.14°C to 27.78°C for all pathogens. Critical limits were established to be used in a HARPC or HACCP plan.ConclusionEstablished critical limits are being used by renderers to comply with FSMA regulations.

Objective This study compared knowledge and food-handling behavior after pathogen-specific (experimental treatment) versus basic food safety instruction (active control) presented during nutrition education classes for low-income English- and Spanish-language pregnant women. Methods Subjects (n = 550) were randomly assigned to treatment groups in two different locations in the United States. Food safety instruction was part of an 8-lesson curriculum. Food safety knowledge and behavior were measured pre/post intervention. Descriptive data were analyzed by Chi-Square or ANOVA; changes after intervention were analyzed by regression analysis. Results Knowledge improved after intervention in the pathogen-specific treatment group compared to active control, especially among Spanish-language women. Behavior change after intervention for the pathogen-specific treatment group improved for thermometer usage, refrigeration and consumption of foods at high risk for safety; however, all other improvements in behavior were accounted for by intervention regardless of treatment group. As expected, higher pre-instruction behavioral competency limited potential gain in behavior post-instruction due to a ceiling effect. This effect was more dominant among English-language women. Improvements were also linked to formal education completed, a partner at home, and other children in the home. Conclusions for Practice This study demonstrated that pathogen-specific food safety instruction leads to enhance knowledge and food handling behaviors that may improve the public health of pregnant women and their unborn children, especially among Spanish-language women. More importantly, food safety instruction, even at the most basic level, benefited pregnant women’s food safety knowledge and food-handling behavior after intervention.

The objective of this study was to identify the maximum time of refrigerated storage before aerobic psychrotrophic bacteria grew to a level indicative of spoilage (7 log cfu/g) or other indicators of spoilage were observed for whole-muscle pork and ground pork sausage packaged using FreshCase technology. Pork chops and pork sausage were packaged using conventional vacuum packaging without nitrite in film (Control) or using FreshCase technology and were compared with respect to microbial counts, pH, instrumental color measurements, lipid oxidation level, and sensory properties. The storage life was 45 d for pork chops stored in FreshCase packages at 1°C and 19 d for ground pork sausage stored under the same condition. Results indicated that both pork chops and sausage stored in FreshCase packages retained redder color ( < 0.05) than those stored in Control packages. No differences ( > 0.05) existed between Control and FreshCase packaged samples for any off-odor detection for either pork chops or sausage. Moreover, levels of oxidative rancidity in all packages had low thiobarbituric acid reactive substances values. The results indicated that FreshCase technology can be used to extend storage life of pork products without having adverse effects on pork quality.

ABSTRACT Escherichia coli O157:H7 is one of the major foodborne pathogens in the United States. We isolated a variant Shiga toxin-negative E. coli O157:H7 strain from feedlot cattle. We report here the draft genome sequence of this isolate, consisting of a chromosome of ~4.8 Mb and two plasmids of ~96 kb and ~14 kb.

Rifampicin-resistant (Rif(R)) strains have often been used in studies of Escherichia coliO157:H7 to assure more specific recovery. The present study compared the heat inactivation kinetics of rifampicin-resistant variants of pathogenic and nonpathogenic E.coliO157 strains with those of the wild-type parental strains. Spontaneous Rif(R) derivatives of 11 pathogenic and six nonpathogenic E.coliO157 strains were selected. Stationary-phase cells of each strain in sterile tryptic soy broth were heated at 60C for 0, 30, 60, 90, 120, 150, 180 or 240s, and aliquots were plated on tryptic soy agar supplemented with 0.1% pyruvate + 100g/mL rifampicin. D values of three pathogenic and two nonpathogenic Rif(R)E.coliO157 strains were 35-70% lower (P<0.05) than those of their wild-type counterparts. However, the heat resistance of most of the Rif(R)E.coliO157:H7 strains (12 strains) was similar (P0.05) to that of their parental counterparts. Therefore, the findings of this study demonstrated that the majority of the rifampicin-resistant E. coli O157:H7 variants that were evaluated to assure more reliable recovery in microbial inactivation studies are suitable for use in various heat challenge studies.Practical ApplicationsMore reliable media are needed for the isolation or recovery of E.coliO157:H7 in inoculated challenge studies. Prior to use of rifampicin-resistant E.coliO157:H7 strains in heat challenge studies, it is important that their thermal inactivation kinetics be compared with those of their wild-type parental strains. The results of this study showed that the thermotolerance at 60C of the majority of the tested rifampicin-resistant E. coliO157:H7 variants was not different than that of the corresponding wild-type strains. These rifampicin-resistant strains can, therefore, be used in heat challenge studies and can be recovered with non-selective culture media supplemented with rifampicin.

The antimicrobial effects of thyme oil (TO), grapefruit seed extract (GSE), and basil essential oil, alone or in combination with cetylpyridinium chloride (CPC), sodium diacetate, or lactic acid, were evaluated against Escherichia coli O157:H7 in a moisture-enhanced beef model system. The model system was composed of a nonsterile beef homogenate to which NaCl (0.5%) and sodium tripolyphosphate (0.25%) were added, together with the tested antimicrobial ingredients. Beef homogenate treatments were inoculated (ca. 3 log CFU/ml) with rifampin-resistant E. coli O157:H7 (eight-strain mixture) and incubated at 15°C (48 h). The most effective individual treatments were TO (0.25 or 0.5%) and GSE (0.5 or 1.0%), which immediately reduced (P < 0.05) pathogen levels by ≥3.4 log CFU/ml. Additionally, CPC (0.04%) reduced initial E. coli O157:H7 counts by 2.7 log CFU/ml. Most combinations of the tested plant-derived extracts with CPC (0.02 or 0.04%) and sodium diacetate (0.25%) had an additive effect with respect to antibacterial activity. In a second study, antimicrobial interventions were evaluated for their efficacy in reducing surface contamination of E. coli O157:H7 on beef cuts and to determine the effect of these surface treatments on subsequent internalization of the pathogen during blade tenderization. Beef cuts (10 by 8 by 3.5 cm) were inoculated (ca. 4 log CFU/g) on one side with the rifampin-resistant E. coli O157:H7 strain mixture and were then spray treated (20 lb/in2, 10 s) with water, GSE (5 and 10%), lactic acid (5%), or CPC (5%). Untreated (control) and spray-treated surfaces were then subjected to double-pass blade tenderization. Surface contamination (4.4 log CFU/g) of E. coli O157:H7 was reduced (P < 0.05) to 3.4 (5% CPC) to 4.1 (water or 5% GSE) log CFU/g following spray treatment. The highest and lowest transfer rates of pathogen cells from the surface to deeper tissues of blade-tenderized sections were obtained in the untreated control and CPC-treated samples, respectively.

Not-ready-to-eat breaded chicken products formulated with antimicrobial ingredients were tested for the effect of sample dimensions, surface browning method and final internal sample temperature on inoculated Salmonella populations. Fresh chicken breast meat portions (5 × 5 × 5 cm), inoculated with Salmonella (7-strain mixture; 5 log CFU/g), were mixed with (5% v/w total moisture enhancement) (i) distilled water (control), (ii) caprylic acid (CAA; 0.0625%) and carvacrol (CAR; 0.075%), (iii) CAA (0.25%) and ε-polylysine (POL; 0.5%), (iv) CAR (0.15%) and POL (0.5%), or (v) CAA (0.0625%), CAR (0.075%) and POL (0.5%). Sodium chloride (1.2%) and sodium tripolyphosphate (0.3%) were added to all treatments. The mixtures were then ground and formed into 9 × 5 × 3 cm (150 g) or 9 × 2.5 × 2 cm (50 g) portions. The products were breaded, browned in (i) an oven (208 °C, 15 min) or (ii) deep fryer (190 °C, 15 s), packaged, and stored at -20 °C (8 d). Overall, maximum internal temperatures of 62.4 ± 4.0 °C (9 × 2.5 × 2 cm) and 46.0 ± 3.0 °C (9 × 5 × 3 cm) were reached in oven-browned samples, and 35.0 ± 1.1 °C (9 × 2.5 × 2 cm) and 31.7 ± 2.6 °C (9 × 5 × 3 cm) in fryer-browned samples. Irrespective of formulation treatment, total (after frozen storage) reductions of Salmonella were greater (P < 0.05) for 9 × 2.5 × 2 cm oven-browned samples (3.8 to at least 4.6 log CFU/g) than for 9 × 5 × 3 cm oven-browned samples (0.7 to 2.5 log CFU/g). Product dimensions did not (P ≥ 0.05) affect Salmonella reductions (0.6 to 2.8 log CFU/g) in fryer-browned samples. All antimicrobial treatments reduced Salmonella to undetectable levels (<0.3 log CFU/g) in oven-browned 9 × 2.5 × 2 cm samples. Overall, the data may be useful for the selection of antimicrobials, product dimensions, and surface browning methods for reducing Salmonella contamination.

Raw drinking milk (RDM) has a diverse microbial flora which can include pathogens transmissible to humans. The main microbiological hazards associated with RDM from cows, sheep and goats, horses and donkeys and camels were identified using a decision tree approach. This considered evidence of milk-borne infection and the hazard being present in the European Union (EU), the impact of the hazard on human health and whether there was evidence for RDM as an important risk factor in the EU. The main hazards were Campylobacter spp., Salmonella spp., shigatoxin-producing Escherichia coli (STEC), Brucella melitensis, Mycobacterium bovis and tick-borne encephalitis virus, and there are clear links between drinking raw milk and human illness associated with these hazards. A quantitative microbiological risk assessment for these hazards could not be undertaken because country and EU-wide data are limited. Antimicrobial resistance has been reported in several EU countries in some of the main bacterial hazards isolated from raw milk or associated equipment and may be significant for public health. Sale of RDM through vending machines is permitted in some EU countries, although consumers purchasing such milk are usually instructed to boil the milk before consumption, which would eliminate microbiological risks. With respect to internet sales of RDM, there is a need for microbiological, temperature and storage time data to assess the impact of this distribution route. Intrinsic contamination of RDM with pathogens can arise from animals with systemic infection as well as from localised infections such as mastitis. Extrinsic contamination can arise from faecal contamination and from the wider farm environment. It was not possible to rank control options as no single step could be identified which would significantly reduce risk relative to a baseline of expected good practice, although potential for an increase in risk was also noted. Improved risk communication to consumers is recommended. (C) European Food Safety Authority, 2015.

Abstract An alternative method to the HTST treatment (High Temperature Short Time pasteurisation at 72 °C for at least 15 seconds or equivalent pasteurisation effect achieving a negative reaction to a phosphatase test), approved for the treatment of bovine colostrum (Category 3 material), was assessed. The purpose of the alternative method, based on a series of filtration steps, is the production of Colostrinov, a product whose main ingredient is bovine colostrum, to be used for foal nutrition. Since the filtration techniques used are known to eliminate particles of the size of bacteria, fungi and protozoa from liquids, it is reasonable to assume that the microfiltration process reduces these contaminants to a level at least equivalent to the treatment required by the legislation. Owing to their small size, viruses are not retained by the mechanical effect of the filters but they may be retained by physico‐chemical interactions with the surface of the filter, depending on the surface properties of the viruses and those of the filter, as well as on the properties of the surrounding liquid. From the information provided by the applicant, it cannot be concluded whether or not the microfiltration process reduces the relevant viral contaminants to a level at least equivalent to a single HTST treatment as required by the legislation.

EFSA is requested to assess the safety of a broad range of biological agents in the context of notifications for market authorisation as sources of food and feed additives, enzymes and plant protection products. The qualified presumption of safety (QPS) assessment was developed to provide a harmonised generic pre-assessment to support safety risk assessments performed by EFSA's scientific Panels. The safety of unambiguously defined biological agents (at the highest taxonomic unit appropriate for the purpose for which an application is intended), and the completeness of the body of knowledge are assessed. Identified safety concerns for a taxonomic unit are, where possible and reasonable in number, reflected as 'qualifications' in connection with a recommendation for a QPS status. A total of 85 biological agents were notified to EFSA between October 2014 and March 2015. From those, 35 biological agents already had a QPS status and were not further evaluated, and 45 were also not included as they are filamentous fungi or enterococci, biological groups which have been excluded from the QPS activities since 2014. Two notifications referred to two taxonomic units which were evaluated for the QPS status, one of which was recommended for the QPS list: Xanthomonas campestris, only for the production of xanthan gum, while the other, Bacillus circulans, was not due to insufficient body of knowledge on a safe history of use in foods and feeds. Three notifications belonging to the genus Streptomyces were not evaluated for the QPS status, because the genus was recently considered not suitable for the QPS approach. (C) European Food Safety Authority, 2015.

Eight tools relevant to risk ranking of biological hazards in food were identified and assessed using two case studies. Differences in their performance were observed, related to the risk metrics, data requirements, ranking approach, model type, model variables and data integration. Quantitative stochastic models are the most reliable for risk ranking. However, this approach needs good characterisation of input parameters. The use of deterministic models that ignore variability may result in risk ranking errors. The ordinal scoring approaches in semi-quantitative models provide ranking with more errors than the deterministic approaches. FDA (Food and Drug Administration)-iRISK was identified as the most appropriate tool for risk ranking of microbiological hazards. The Burden of Communicable Diseases in Europe (BCoDE) toolkit can be used in combination with the outputs from FDA-iRISK or as a top-down tool to rank pathogens. Uncertainty needs to be addressed and communicated to decision makers and stakeholders as one of the outcomes of the risk ranking process. Uncertainty and variability can be represented by means of probability distributions. Techniques such as the NUSAP (numeral, unit, spread, assessment and pedigree) approach can also be used to prioritise factors for sensitivity and scenario analysis or stochastic modelling. Quantitative risk ranking models are preferred over semi-quantitative models. When data and time constraints do not allow quantitative risk ranking, semiquantitative models could be used, but the limitations of these approaches linked to the selection and integration of the ordinal scores should be made explicit. Decision trees should be used only to show how decisions are made about classifying food-pathogen combinations into broad categories. BCoDE and FDA-iRISK, in combination with a network of available predictive microbiology tools, databases and information sources, can form a risk ranking toolbox and be applied based on a "fit for purpose" approach supporting timely and transparent risk ranking. (C) European Food Safety Authority, 2015.

Microbial contamination of meat and meat products is unavoidable as microorganisms are present on animals and in their environment. Thus, raw and not fully heated (commercially processed) or otherwise processed/preserved (e.g. frozen, fermented/dried, high hydrostatic pressure processed, irradiated) meat and meat products are prone to spoilage and compromised safety due to microbial presence and growth. Raw meat products (although few consumers eat certain meat products raw or undercooked, intentionally or accidentally; a practice not recommended) need further processing and/or cooking before consumption. This makes them shelf-stable or semi-perishable, and safe for consumption or ready-to-eat. In general, the shelf-life, quality and safety of meat and meat products are extended and improved through adequate processing, appropriate marketing, storage and preparation for consumption, under properly clean, sanitary and hygienic conditions, following an integrated approach throughout all sectors of the food supply web, including producers, processors, distributors, retailers, as well as consumers. The strategy for hazard control should include: (1) good animal production practices on the farm; (2) slaughtering of animals that are disease-free; (3) processing of carcasses and meat in properly designed and maintained facilities and under sanitary and hygienic conditions; (4) use of decontamination intervention strategies, if approved, to reduce microbial levels when needed; (5) thermal processing, freezing, drying, fermentation, acidification, use of approved antimicrobials in certain products, and packaging; (6) maintenance of proper cold chain conditions during distribution; (7) proper storage and preparation procedures by food service and consumers; and (8) management of every segment of this common sense but complex system, with well-validated, verified and documented programs such as the hazard analysis critical control point (HACCP) system.

The Qualified Presumption of Safety (QPS) approach, initially developed for the assessment of microorganisms referred to EFSA and added to the food chain is equally applicable to the assessment of botanicals or botanical preparations. Using the principles to establish the suitability of a botanical preparation for QPS status, it has been possible to develop a structured assessment scheme that provides a practical method for assessing botanicals and botanical preparations for which an adequate body of knowledge exists and therefore without the need for further testing. Reiterative applications of the assessment scheme to related botanicals or different botanical preparations obtained from the same plant variety can allow a QPS status to be derived for specific groupings. However, the particularity of botanicals that may be presented in a wide variety of forms or whose morphology and chemical composition may be markedly affected by geographical and environmental factors, makes the possibility to establish QPS status at high taxonomic levels quite limited. Still, the above-mentioned structured approach for the assessment of botanicals and botanical preparations represents a considerable advancement in the development of a comprehensive, systematic and transparent methodology. The Scientific Committee recommends its use as an extension of the 2009 EFSA guidance for the safety assessment of botanicals and botanical preparations intended to be used in food supplements. (-)-Synephrine has also been detected by Inafuku-Teramoto et al. (2011) in the peel of the fruits of all the analysed Citrus species: C. depressa, C. madurensis, C. rokugatsu, C. oto, C. keraji, C. nobilis, C. tankan, C. tangerine.

Studies evaluating the safety and efficacy of solutions, containing peroxyacetic acid (PAA) as the active ingredient, in mixtures with acetic acid, hydrogen peroxide, and 1-hydroxyethylidene-1,1-diphosphonic acid (HEDP) and possibly octanoic acid and peroxyoctanoic acid, for reduction of pathogens on poultry carcasses and meat were assessed. Treatments at ambient temperature consisted of dipping in short term baths, in chiller baths or spraying. On the basis of the previous EFSA exposure scenarios including short term baths that were not evaluated previously, no toxicity concerns were identified with regard to residues of peroxyacids, to HEDP and to possible reaction products of hydrogen peroxide and peroxyacids with lipids and proteins of the poultry carcasses. A relevant reduction of PAA treatment on E. coli and coliforms was demonstrated by dipping warm carcasses, but few data were available for pathogens (Salmonella and Campylobacter). Spraying appeared to be less effective than dipping in reducing indicator organisms than dipping. When dipping chilled carcasses, reduction of indicator organisms and pathogens was evident, although only in low or medium strength of evidence studies. In chiller bath application, there was a relevant impact on E. coli, but less effect on coliforms, and little data was available on reduction of pathogens. The emergence of acquired reduced susceptibility to biocides and/ or resistance to therapeutic antimicrobials following the use of PAA was considered unlikely. There were no concerns for environmental risk of peroxyacids, acetic acid and octanoic acid. On the basis of a conservative preliminary guideline for surface water quality, the emission of HEDP from a poultry plant into the environment could not be considered safe a priori. It was recommended that HACCP plans should include monitoring of the concentration of HEDP and of the decontaminating substance in the working solution and post-marketing surveillance for resistance in both pathogenic and commensal bacteria. (C) European Food Safety Authority, 2014

red meat, vacuum packed beef and poultry could be stored at 2 °C for up to 14, 39 and 5 days, respectively, without more bacterial pathogen growth occurring than that which would be achieved under current legislative conditions. It was therefore concluded that alternative time-temperature combinations for the storage of fresh meat between slaughter and mincing are possible without increasing bacterial pathogen growth, and maximum times for the storage of fresh meat intended for minced meat preparation are provided for different storage temperatures. The impact of spoilage on maximum storage times was not considered

For generations, those that produce livestock and meat generally felt that their country or geographical region (i.e., provenance) reflected a basis for product differentiation. This occurs to the extent that geography of production often is considered a “brand.” For example, there exists “U.S. Grain-Fed Beef” or “Kobe Black Wagyu” or “Uruguayan Grass-Fed Lamb” or “Danish Pork.” However, for most meat trade, industry has evolved beyond this. With the exception perhaps of farms onto which livestock are born, meat company's profits are not generally tied to geographical considerations. Most major companies (e.g., JBS, Marfrig, Tyson, Cargill, Danish Crown, Nippon Meat Packers, etc.) operate in multiple countries and represent to consumers the production of a number of locations. However, there also now exist entrepreneurial options for meat production and “local” sales, albeit at lesser volumes. This discussion explores “global” and “local” meat marketing options.

member and chair of the EFSA Panel on Food Contact Materials, Enzymes, Flavourings and Processing Aids (CEF) passed away suddenly on