Nicholas J. Sevart

Evaluating the Efficacy of Three U.S. Department of Agriculture–Approved Antimicrobial Sprays for Reducing Shiga Toxin–Producing Escherichia coli Surrogate Populations on Bob Veal Carcasses

Effective antimicrobial intervention strategies to reduce Shiga toxin-producing Escherichia coli (STEC) risks associated with veal are needed. This study evaluated the efficacy of lactic acid (4.5%, pH 2.0), Citrilow (pH 1.2), and Beefxide (2.25%, pH 2.3) for reducing STEC surrogates on prerigor and chilled bob veal carcasses and monitored the effects of these interventions on chilled carcass color. Dehided bob veal carcasses were inoculated with a five-strain cocktail of rifampin-resistant, surrogate E. coli bacteria.E. coli surrogates were enumerated after inoculation, after water wash, after prechill carcass antimicrobial spray application, after chilling for 24 h, and after postchill carcass antimicrobial spray application; carcass color was measured throughout the process. A standard carcass water wash (∼50°C) reduced the STEC surrogate population by 0.9 log CFU/cm(2) (P ≤ 0.05). All three antimicrobial sprays applied to prerigor carcasses delivered an additional ∼0.5-log reduction (P ≤ 0.05) of the surrogates. Chilling of carcasses for 24 h reduced (P ≤ 0.05) the surrogate population by an additional ∼0.4 log cycles. The postchill application of the antimicrobial sprays provided no further reductions. Carcass L*, a*, and b* color values were not different (P > 0.05) among carcass treatments. Generally, the types and concentrations of the antimicrobial sprays evaluated herein did not negatively impact visual or instrumental color of chilled veal carcasses. This study demonstrates that warm water washing, followed by a prechill spray treatment with a low-pH chemical intervention, can effectively reduce STEC risks associated with veal carcasses; this provides processors a validated control point in slaughter operations.

Validation of Baking To Control Salmonella Serovars in Hamburger Bun Manufacturing, and Evaluation of Enterococcus faecium ATCC 8459 and Saccharomyces cerevisiae as Nonpathogenic Surrogate Indicators.

This study was conducted to validate a simulated commercial baking process for hamburger buns to destroy Salmonella serovars and to determine the appropriateness of using nonpathogenic surrogates (Enterococcus faecium ATCC 8459 or Saccharomyces cerevisiae) for in-plant process validation studies. Wheat flour was inoculated (∼6 log CFU/g) with three Salmonella serovars (Typhimurium, Newport, or Senftenberg 775W) or with E. faecium. Dough was formed, proofed, and baked to mimic commercial manufacturing conditions. Buns were baked for up to 13 min in a conventional oven (218.3°C), with internal crumb temperature increasing to ∼100°C during the first 8 min of baking and remaining at this temperature until removal from the oven. Salmonella and E. faecium populations were undetectable by enrichment (>6-log CFU/g reductions) after 9.0 and 11.5 min of baking, respectively, and ≥5-log-cycle reductions were achieved by 6.0 and 7.75 min, respectively. D-values of Salmonella (three-serovar cocktail) and E. faecium 8459 in dough were 28.64 and 133.33, 7.61 and 55.67, and 3.14 and 14.72 min at 55, 58, and 61°C, respectively, whereas D-values of S. cerevisiae were 18.73, 5.67, and 1.03 min at 52, 55, and 58°C, respectivly. The z-values of Salmonella, E. faecium, and S. cerevisiae were 6.58, 6.25, and 4.74°C, respectively. A high level of thermal lethality was observed for baking of typical hamburger bun dough, resulting in rapid elimination of high levels of the three-strain Salmonella cocktail; however, the lethality and microbial destruction kinetics should not be extrapolated to other bakery products without further research. E. faecium demonstrated greater thermal resistance compared with Salmonella during bun baking and could serve as a conservative surrogate to validate thermal process lethality in commercial bun baking operations. Low thermal tolerance of S. cerevisiae relative to Salmonella serovars limits its usefulness as a surrogate for process validations.

Electrostatic spray cabinet evaluation to verify uniform delivery of chemical and biological solutions to pre-chilled meat animal carcasses

Introduction Shiga toxin-producing Escherichia coli (STEC) are a group of bacteria that cause an estimated 265,000 illnesses, 3,600 hospitalizations, and 30 deaths annually in the United States. STEC are frequently associated with raw or undercooked meat products, prompting the beef industry to develop and apply various antimicrobial intervention technologies during processing operations. The application of chemical antimicrobials to carcasses and fabricated cuts using an electrostatic spray (ESS) system (Figure 1) offers several potential advantages for controlling disease-causing pathogens, including enhanced chemical deposition (coverage) profiles, reduced overspray wastage of foodgrade antimicrobials, and reduced water requirements. The objectives of this study were to (1) calibrate an ESS carcass cabinet installed at the Kansas State University Biosecurity Research Institute, (2) test the chemical deposition profile of the ESS cabinet onto a meat carcass using fluorescent dye, and (3) determine if the ESS could be used to uniformly apply a biological inoculum to a carcass to support pathogen-inoculated validation studies of different chemical intervention technologies to support the needs of the beef processing industry.

Use of an Electrostatic Spraying System or the Sprayed Lethality in Container Method To Deliver Antimicrobial Agents onto the Surface of Beef Subprimals To Control Shiga Toxin–Producing Escherichia coli

The efficacy of an electrostatic spraying system (ESS) and/or the sprayed lethality in container (SLIC) method to deliver antimicrobial agents onto the surface of beef subprimals to reduce levels of Shiga toxin-producing Escherichia coli (STEC) was evaluated. Beef subprimals were surface inoculated (lean side; ca. 5.8 log CFU per subprimal) with 2 mL of an eight-strain cocktail comprising single strains of rifampin-resistant (100 μg/mL) STEC (O26:H11, O45:H2, O103:H2, O104:H4, O111:H-, O121:H19, O145:NM, and O157:H7). Next, inoculated subprimals were surface treated with lauric arginate (LAE; 1%), peroxyacetic acid (PAA; 0.025%), or cetylpyridinium chloride (CPC; 0.4%) by passing each subprimal, with the inoculated lean side facing upward, through an ESS cabinet or via SLIC. Subprimals were then vacuum packaged and stored at 4°C. One set of subprimals was sampled after an additional 2 h, 3 days, or 7 days of refrigerated storage, whereas another set was retreated via SLIC after 3 days of storage with a different one of the three antimicrobial agents (e.g., a subprimal treated with LAE on day 0 was then treated with PAA or CPE on day 3). Retreated subprimals were sampled after 2 h or 4 days of additional storage at 4°C. A single initial application of LAE, PAA, or CPC via ESS or SLIC resulted in STEC reductions of ca. 0.3 to 1.3 log CFU per subprimal after 7 days of storage. However, when subprimals were initially treated with LAE, PAA, or CPC via ESS or SLIC and then separately retreated with a different one of these antimicrobial agents via SLIC on day 3, additional STEC reductions of 0.4 to 1.0 log CFU per subprimal were observed after an additional 4 days of storage. Application of LAE, PAA, or CPC, either alone or in combination, via ESS or SLIC is effective for reducing low levels (ca. 0.3 to 1.6 log CFU) of STEC that may be naturally present on the surface of beef subprimals.

Validation of a Sequential Hide-On Bob Veal Carcass Antimicrobial Intervention Composed of a Hot Water Wash and Lactic Acid Spray in Combination with Scalding To Control Shiga Toxin–Producing Escherichia coli Surrogates†

Scalding of hide-on bob veal carcasses with or without standard scalding chemical agents typically used for hogs, followed by an 82.2°C hot water wash and lactic acid spray (applied at ambient temperature) before chilling, was evaluated to determine its effectiveness in reducing Shiga toxin-producing Escherichia coli surrogate populations. A five-strain cocktail of rifampin-resistant, nonpathogenic E. coli surrogates was used to inoculate hides of veal carcasses immediately after exsanguination (target inoculation level of 7.0 log CFU/100 cm2). For carcasses receiving no scalding treatments, spraying with 82.2°C water as a final wash resulted in a 4.5-log CFU/100 cm2 surrogate reduction, and an additional 1.2-log CFU/100 cm2 reduction was achieved by spraying with 4.5% lactic acid before chilling. Scalding hide-on carcasses in 60°C water (no chemicals added) for 4 min in a traditional hog scalding tank resulted in a 2.1-log CFU/100 cm2 reduction in surrogate levels, and a subsequent preevisceration 82.2°C water wash provided an additional 2.9-log CFU/100 cm2 reduction. Spraying a 4.5% solution of lactic acid onto scalded, hide-on carcasses (after the 82.2°C water wash) resulted in a minimal additional reduction of 0.4 log CFU/100 cm2. Incorporation of scalding chemicals into the scald water resulted in a 4.1-log CFU/100 cm2 reduction (1.9 log CFU/100 cm2 greater than scalding without chemicals) in the surrogate population, and the first 82.2°C wash provided an additional 2.5-log CFU/100 cm2 reduction. Application of antimicrobial interventions did not affect the carcass temperature decline during chilling, the pH decline, or the color characteristics of the ribeye or the flank of the bob veal carcasses.

SHIGA TOXIN-PRODUCING ESCHERICHIA COLI IN MEAT: A PRELIMINARY SIMULATION STUDY ON DETECTION CAPABILITIES FOR THREE SAMPLING METHODS

Contamination by Shiga Toxin-producing Escherichia coli (STEC) is a continuing concern for meat production facility management throughout the United States. Several methods have been used to detect STEC during meat processing, however the excessive experimental cost of determining the optimal method is rarely feasible. The objective of this preliminary simulation study is to determine which sampling method (Cozzini core sampler, core drill shaving, and N-60 surface excision) will better detect STEC at varying levels of contamination present in the meat. 1000 simulated experiments were studied using a binary model for rare occurrences to find the optimal method. We found that for meat contamination levels less than 0.1% or greater than 10% all sampling methods perform equally. At moderate levels of contamination (between 0.1% and 10%) core drill shaving and N-60 perform significantly better than Cozzini core sampler. However, there does not appear to be a significant difference between core drill shaving and N60. This project was supported by an Agriculture and Food Research Initiative Competitive Grant no. 2012-68003-30155 from the USDA National Institute of Food and Agriculture.

Effects of media type on Shiga toxigenic E. coli growth patterns

Introduction Escherichia coli O157:H7 was declared to be an adulterant in raw ground beef in 1994 by the United States Department of Agriculture Food Safety and Inspection Service following a large and deadly foodborne disease outbreak in the Pacific Northwest involving undercooked hamburgers sold at Jack-in-the-Box restaurants. Due to their recognition as significant human foodborne pathogens, six additional strains (serotypes) of Shiga toxin-producing E. coli (STEC) were also deemed to be adulterants in raw beef products in 2012.