Tulane Virus Persistence and Microbial Stability in 3D Food Ink under Various Storage Conditions: A Pre- and Post-Printing Analysis.

3D food printers facilitate novel customization of the physicochemical properties of food. This study aimed to investigate the impact of storage conditions on the inactivation of the human norovirus surrogate, Tulane virus (TuV), within 3D printed foods. TuV-inoculated protein cookie food ink (∽ 4 log PFU/g) was distributed into 18 3D food printer capsules (50 g each); half immediately underwent extrusion. Storage of the capsules and printed food products at 20 °C (0, 6, 12, and 24 h), 4 °C (0, 1, 3, and 5d), and - 18 °C (0, 1, 3, and 5d) was completed before analysis for TuV via plaque assays in addition to aerobic plate count, yeast and mold counts, and pH and water activity (aw) measurements. A significant 3-way interaction effect was observed between time, temperature, and storage method (capsule/print) (p = 0.006). Significant findings include: (1) A greater reduction in virions was observed in capsules after 24 h at 20 °C and (2) a substantial reduction in virions at 4 °C from day 0 to day 1 was observed, independent of storage method. Microbial indicators remained steady across temperatures, with storage temperature significantly impacting pH and aw. A significant two-way interaction effect (p = 0.006) was found between microorganism type (yeast/aerobic counts) and temperature. This research seeks to provide insights for the food industry and regulatory bodies in crafting guidelines for the safe storage and handling of 3D printed foods and inks.

From bytes to bites: Advancing the food industry with three-dimensional food printing.

The rapid advancement of three-dimensional (3D) printing (i.e., a type of additive manufacturing) technology has brought about significant advances in various industries, including the food industry. Among its many potential benefits, 3D food printing offers a promising solution to deliver products meeting the unique nutritional needs of diverse populations while also promoting sustainability within the food system. However, this is an emerging field, and there are several aspects to consider when planning for use of 3D food printing for large-scale food production. This comprehensive review explores the importance of food safety when using 3D printing to produce food products, including pathogens of concern, machine hygiene, and cleanability, as well as the role of macronutrients and storage conditions in microbial risks. Furthermore, postprocessing factors such as packaging, transportation, and dispensing of 3D-printed foods are discussed. Finally, this review delves into barriers of implementation of 3D food printers and presents both the limitations and opportunities of 3D food printing technology.

Cultivating Food Safety Together: Insights About the Future of Produce Safety in the U.S. Controlled Environment Agriculture Sector.

Controlled environment agriculture (CEA) is a rapidly growing sector that presents unique challenges and opportunities in ensuring food safety. This manuscript highlights critical gaps and needs to promote food safety in CEA systems as identified by stakeholders (n=47) at the Strategizing to Advance Future Extension andResearch (S.A.F.E.R.) CEA conference held in April 2023 at The Ohio State University's Ohio CEA Research Center. Feedback collected at the conference was analyzed using an emergent thematic analysis approach to determine key areas of focus. Research-based guidance is specific to the type of commodity, production system type, and size. Themes include the need for improved supply chain control, cleaning, and sanitization practices, pathogen preventive controls and mitigation methods and training and education. Discussions surrounding supply chain control underscored the significance of the need for approaches to mitigate foodborne pathogen contamination. Effective cleaning and sanitization practices are vital to maintaining a safe production environment, with considerations such as establishing standard operating procedures, accounting for hygienic equipment design, and managing the microbial communities within the system. Data analysis further highlights the need for risk assessments, validated pathogen detection methods, and evidence-based guidance in microbial reduction. In addition, training and education were identified as crucial in promoting a culture of food safety within CEA. The development of partnerships between industry, regulatory, and research institutions are needed to advance data-driven guidance and practices across the diverse range of CEA operations and deemed essential for addressing challenges and advancing food safety practices in CEA. Considering these factors, the CEA industry can enhance food safety practices, foster consumer trust, and support its long-term sustainability.

Aqueous Ozone Efficacy for Inactivation of Foodborne Pathogens on Vegetables Used in Raw Meat-Based Diets for Companion Animals.

The present study evaluates the efficacy of a batch wash ozone sanitation system (BWOSS) and spray wash ozone sanitation system (SWOSS) against Listeria monocytogenes (two strains) and Salmonella enterica subsp. enterica (three serovars) inoculated on the surface of carrots, sweet potatoes, and butternut squash, commonly used in raw meat-based diets (RMBDs) marketed for companion animals such as dogs and cats. Produce either remained at room temperature for 2 h or were frozen at -20°C and then tempered overnight at 4°C to mimic the preprocessing steps of a raw pet food processing operation ('freeze-temper') prior to ozone treatment. Two ozone concentrations (0 and 5 ppm) were applied for either 20 s or 60 s for BWOSS and 20 s for SWOSS. Based on an ANOVA, BWOSS data showed no significant difference (P > 0.05) in microbial reduction between 0 and 5 ppm ozone concentration across all treatment durations for each produce type. BWOSS resulted in mean microbial reductions of up to 1.56 log CFU/mL depending on the treatment time and produce type. SWOSS data were analyzed using a generalized linear model with Quasipoisson errors. Freeze-tempered produce treated with SWOSS had a higher bacterial log reduction at 5 ppm ozone compared to 0 ppm ozone (P = 0.0013) whereas room temperature produce treated with SWOSS did not show any significant difference in microbial reduction between ozone concentrations. The potential to mitigate microbial cross-contamination was also investigated during SWOSS treatment. The results indicate that 5 ppm ozone decreased pathogens in the rinsate and proximal surfaces by 0.63-1.66 log CFU/mL greater than no ozone depending on the pathogen and sample. Overall, data from this study indicate that SWOSS would be more effective compared to BWOSS in reducing the microbial load present on the surface of root tubers and squash subjected to freezing and thawing and has the potential to mitigate cross-contamination within RMDB manufacturing environments.

Growing Safer Greens: Exploring Food Safety Practices and Challenges in Indoor, Soilless Production Through Thematic Analysis of Leafy Greens Grower Interviews.

Indoor, soilless production-often referred to more broadly as controlled environment agriculture (CEA)-is increasingly used for the cultivation of leafy greens. Minimal information is currently available regarding food safety practices during production and distribution of leafy greens grown within indoor, soilless environments in the United States (U.S.). This study aimed to describe production challenges and implementation of good agricultural practices among CEA growers. Data collection methods included semi-structured interviews (N = 25) and a supplemental online survey completed by growers (N = 12) in the U.S. Out of 18 total responses (i.e., multiple responses allowed per completed survey), survey data indicated that lettuce (n = 5; 27.8%) was the most commonly grown leafy green, followed by culinary herbs (n = 3; 16.7%) and arugula (n = 3; 16.7%). Most growers (n = 7; 58.3%) grew other agricultural products, specifically other crops in addition to leafy greens. Revenue from sales ranged from US$500 000 per year. Meanwhile, nearly half (n = 5; 45.5%) of respondents (N = 11) were uncertain whether their produce was subject to the FSMA Produce Safety Rule. Most survey respondents used vertical farming techniques (5 out of 11; 45.5%) or some variety of greenhouse (4 out of 11; 36.4%). Based on 35 total responses, leafy greens were most commonly sold to "Commercial Restaurants" (n = 7; 20.0%), "Grocery Stores" (n = 7; 20.0%), "Institutional Foodservice Establishments (hospitals, schools, childcare, long-term care)" (n = 6; 17.1%), and "Wholesaler/Distributers" (n = 6; 17.1%). The 11 interview questions elucidated three major themes: contextual, barriers to risk management and regulatory compliance, and research needs. Thirteen subthemes were identified, and an example of a subtheme within each major theme, respectively, includes worker hygiene and training, regulatory and certification environment, and risk assessments of individual issues.

Transfer rates of Salmonella Typhimurium, Listeria monocytogenes, and a human norovirus surrogate impacted by macronutrient composition of food inks in 3D food printing systems.

3D food printers (3DFPs) allow for the customization of the physiochemical properties of foods in new ways. Transfer kinetics of foodborne pathogens between surfaces and food inks have not been evaluated in 3DFPs. This study aimed to determine if the macromolecular composition of food inks would impact the transfer rate of foodborne pathogens from the stainless steel food ink capsule to the 3D printed food. Salmonella Typhimurium, Listeria monocytogenes, and a human norovirus surrogate, Tulane virus (TuV), were inoculated onto the interior surface of stainless steel food ink capsules and dried for 30 min. Subsequently, 100 g of one of the following prepared food inks was extruded: (1) pure butter, (2) a powdered sugar solution, (3) a protein powder solution, and (4) a 1:1:1 ratio of all three macromolecules. Pathogen enumeration was completed for both the soiled capsules and the printed food products and resulting transfer rates were estimated using a generalized linear model with quasibinomial errors. A significant two-way interaction effect was found between microorganism type and food ink type (P = 0.0002). Tulane virus was typically transferred the most, and no significant differences between L. monocytogenes and S. Typhimurium were observed for any food matrix or across food matrices. Among food matrices, the complex mixture transferred fewer microorganisms in all instances, while butter, protein, and sugar were all statistically indistinguishable. This research seeks to further develop the field of 3DFP safety and to advance the understanding of the role of macromolecular composition in pathogen transfer kinetics, which have not previously been explored in pure matrices.

Efficacy of Manufacturer Recommendations for the Control of Salmonella Typhimurium and Listeria monocytogenes in Food Ink Capsules Utilized in 3D Food Printing Systems.

The adoption of 3D food printing systems has allowed for the personalization of food properties such as color, shape, and texture. This study aimed to determine if manufacturer cleaning recommendations for stainless steel food ink capsules utilized in 3D food printers adequately control foodborne pathogens of concern, as the recommendations have not been tested. A cocktail of ∼9 log10 CFU/mL each of Salmonella Typhimurium and Listeria monocytogenes was inoculated onto the interior surface of the capsules. Capsules were either unsoiled or soiled with one of the following: butter, protein powder solution, powdered sugar solution, or a mixture containing all three food components. The prepared capsules underwent one of three hygienic protocols: manual washing (MW), a dishwasher speed cycle (DSC), or a dishwasher heavy cycle (DHC). The interaction effect between DSC and the soil mixture was significant (P = 0.01), with the combination achieving an estimated mean log reduction of 5.28 (95% CI: 4.61, 6.05) for L. monocytogenes and 6.69 (95% CI: 6.03, 7.41) for S. Typhimurium. The DSC was the least effective method of cleaning when compared with MW and the DHC. No significant differences were found by placing capsules on the right or left side of the dishwasher (P > 0.1). The interaction effect between wash type and capsule position was significant (P = 0.0007), with the soil mixture and DSC combination achieving an estimated mean log reduction of 3.48 (95% CI: 2.72, 4.45) for the front-most position versus 7.92 (95% CI: 6.72, 9.31) for the back-most position. Soil matrix, cleaning protocol, and capsule position all significantly impact capsule cleanability and therefore food safety risk. The DHC is recommended, and the corners should be avoided during dishwasher loading. The current study provides practical information for consumers, restaurants, industry, and regulatory industries regarding the best practices for cleaning 3D food printers.

Surface Inactivation of a SARS-CoV-2 Surrogate with Hypochlorous Acid is Impacted by Surface Type, Contact Time, Inoculum Matrix, and Concentration.

Indirect contact with contaminated surfaces is a potential transmission route for COVID-19. Therefore, it is necessary to investigate convenient and inexpensive surface sanitization methods, such as HOCl, against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The SARS-CoV-2 surrogate, Phi6 (~ 7 log PFU/mL), was prepared in artificial saliva and tripartite matrices, spot inoculated on coupons of either stainless steel or vinyl, and allowed to dry. The coupons were sprayed with either 500 ppm or 1000 ppm HOCl, and remained on the surface for 0 s (control), 5 s, 30 s, or 60 s. Samples were enumerated via the double agar overlay assay. Statistical analysis was completed in R using a generalized linear model with Quasipoisson error approximations. Time, concentration, surface type, and inoculum matrix were all significant contributors to log reduction at P = 0.05. Significant three-way interactions were observed for 1000 ppm, vinyl, and 60 s (P = 0.03) and 1000 ppm, tripartite, and 60 s (P = 0.0121). A significant two-way interaction between vinyl and 60 s was also observed (P = 0.0168). Overall, increased HOCl concentration and exposure time led to increased Phi6 reduction. Notably, the highest estimated mean log reduction was 3.31 (95% CI 3.14, 3.49) for stainless steel at 60 s and 1000 ppm HOCl in artificial saliva, indicating that this method of sanitization may not adequately reduce enveloped viruses to below infective thresholds.

Performance of Manufacturer Cleaning Recommendations Applied to 3D Food Ink Capsules for the Control of a Human Norovirus Surrogate.

With the widespread availability of 3D food printing systems for purchase, users can customize their food in new ways. Manufacturer recommendations for cleaning these machines remain untested with regard to the prevention of foodborne pathogen transmission. This study aimed to determine if manufacturer cleaning recommendations for food ink capsules utilized in 3D food printers are adequate to control human norovirus (HuNoV). A HuNoV surrogate, Tulane virus (TuV; ~ 6 log10 PFU/mL), was inoculated onto the interior surface of stainless steel food ink capsules. Capsules were either unsoiled or soiled with one of the following: butter, protein powder solution, powdered sugar solution, or a mixture containing all three food components. The capsules were allowed to dry and then one of three hygienic protocols was applied: manual washing (MW), a dishwasher speed cycle (DSC), or a dishwasher heavy cycle (DHC). The interaction effect between DSC and pure butter was a significant predictor of log reduction (P = 0.0067), with the pure butter and DSC combination achieving an estimated mean log reduction of 4.83 (95% CI 4.13, 5.59). The DSC was the least effective method of cleaning when compared with MW and the DHC. The 3-way interaction effects between wash type, soil, and capsule position were a significant predictor of log reduction (P = 0.00341). Capsules with butter in the DSC achieved an estimated mean log reduction of 2.81 (95% CI 2.80, 2.83) for the front-most position versus 6.35 (95% CI 6.33, 6.37) for the back-most position. Soil matrix, cleaning protocol, and capsule position all significantly impact capsule cleanability and potential food safety risk. The DHC is recommended for all capsules, and the corners should be avoided when placing capsules into the dishwasher. The current study seeks to provide recommendations for users of additive manufacturing and 3D food printing including consumers, restaurants, industry, and regulatory industries.

Transfer of Phi6 bacteriophage between human skin and surfaces common to consumer-facing environments.

AIMS: This study aimed to determine the extent of Phi6 (Φ6) transfer between skin and surfaces relevant to consumer-facing environments based on inoculum matrix, surface type and contact time. METHODS AND RESULTS: Φ6 transfer rates were determined from skin-to-fomite and fomite-to-skin influenced by inoculum matrix (artificial saliva and tripartite), surface type (aluminium, plastic, stainless steel, touchscreen, vinyl and wood) and contact time (5 and 10 s). Significant differences in estimated means were observed based on surface type (both transfer directions), inoculum matrix (skin-to-fomite) and contact time (both transfer directions). During a sequential transfer experiment from fomite-to-skin, the maximum number of consecutive transfer events observed was 3.33 ± 1.19, 2.33 ± 1.20 and 1.67 ± 1.21 for plastic, touchscreen and vinyl, respectively. CONCLUSIONS: Contact time significantly impacted Φ6 transfer rates, which may be attributed to skin absorption dynamics. Surface type should be considered for assessing Φ6 transfer rates. SIGNIFICANCE AND IMPACT OF THE STUDY: Although the persistence of Φ6 on fomites has been characterized, limited data are available regarding the transfer of Φ6 among skin and fomites. Determining Φ6 transfer rates for surfaces in consumer-facing environments based on these factors is needed to better inform future virus transmission mitigation strategies.