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.

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.

Factors Impacting Persistence of Phi6 Bacteriophage, an Enveloped Virus Surrogate, on Fomite Surfaces.

The persistence of Phi6 (Φ6) bacteriophage on surfaces commonly encountered in consumer-facing environments was evaluated. Φ6 has been utilized as a surrogate for enveloped viruses, including SARS-CoV-2-the causative agent of COVID-19-due to structural similarities, biosafety level 1 (BSL-1) status, and ease of use. Φ6 persistence on fomites was evaluated by characterizing the impact of the inoculum matrix (artificial saliva, phosphate-buffered saline [PBS], tripartite), inoculum level (low and high), and surface type (nonporous-aluminum, stainless steel, plastic, touchscreen, vinyl; porous-wood). Φ6 was inoculated onto surfaces at low and high inoculum levels for each inoculum matrix and incubated (20.54 ± 0.48°C) for up to 168 h. Φ6 was eluted from the surface and quantified via the double agar overlay assay to determine virus survival over time. For nonporous surfaces inoculated with artificial saliva and PBS, significantly higher D values were observed with high inoculum application according to the 95% confidence intervals. In artificial saliva, D values ranged from 1.00 to 1.35 h at a low inoculum and 4.44 to 7.05 h at a high inoculum across inoculation matrices and surfaces. D values for Φ6, regardless of the inoculum level, were significantly higher in tripartite than in artificial saliva and PBS for nonporous surfaces. In contrast with artificial saliva or PBS, D values in tripartite at low inoculum (D values ranging from 45.8 to 72.8 h) were greater than those at high inoculum (D values ranging from 26.4 to 45.5 h) on nonporous surfaces. This study characterized the impact of the inoculum matrix, inoculum level, and surface type on Φ6 survival on various surfaces relevant to fomite transmission in public settings. IMPORTANCE An important consideration in virus contact transmission is the transfer rate between hands and surfaces, which is driven by several factors, including virus persistence on inanimate surfaces. This research characterized Φ6 persistence on surfaces commonly encountered in public settings based on various factors. The inoculum matrix, which simulates the route of transmission, can impact virus persistence, and three separate matrices were evaluated in this study to determine the impact on Φ6 persistence over time. The number of microorganisms has also been suggested to impact persistence, which was evaluated here to simulate real-world contamination scenarios on six surface types. Results from this study will guide future research utilizing Φ6 or other surrogates for enveloped viruses of public health concern.

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.

Pathogen and Surrogate Survival in Relation to Fecal Indicator Bacteria in Freshwater Mesocosms.

The microbial quality of agricultural water for fresh produce production is determined by the presence of the fecal indicator bacterium (FIB) Escherichia coli, despite poor correlations with pathogen presence. Additional FIB, such as enterococci, have been utilized for assessing water quality. The study objective was to determine the survival times (first time to detect zero or censored) of FIB (E. coli and enterococci), surrogates (Listeria innocua, Listeria seeligeri, Salmonella enterica serovar Typhimurium, and PRD1), and pathogens (four strains each of pathogenic E. coli and Listeria monocytogenes and five Salmonella serovars) simultaneously inoculated in freshwater mesocosms exposed to diel and seasonal variations. Six separate mesocosm experiments were conducted for ≤28 days each season, with samples (sediment/water) collected each day for the first 7 days and weekly thereafter. Microorganisms survived significantly longer in sediment than in water (hazard ratio [HR] for water/sediment is 2.2; 95% confidence interval [CI], 1.79 to 2.71). Also, FIB E. coli survived significantly longer than FIB enterococcus (HR for enterococci/E. coli is 12.9 [95% CI, 8.18 to 20.37]) after adjusting for the sediment/water and lake/river effects. Differences in the area under the curve (calculated from log CFU or PFU over time) were used to assess pathogen and surrogate survival in relation to FIB. Despite sample type (sediment/water) and seasonal influences, survival rates of pathogenic Salmonella serovars were similar to those of FIB E. coli, and survival rates of L. monocytogenes and pathogenic E. coli were similar to those of FIB enterococci. Further investigation of microbial survival in water and sediment is needed to determine which surrogates are best suited to assess pathogen survival in agricultural water used in irrigation water for fresh produce. IMPORTANCE Contamination of fresh produce via agricultural water is well established. This research demonstrates that survival of fecal indicator bacteria, pathogenic microorganisms, and other bacterial and viral surrogates in freshwater differs by sample type (sediment/water) and season. Our work highlights potential risks associated with pathogen accumulation and survival in sediment and the possibility for resuspension and contamination of agricultural water used in fresh produce production. Specifically, a greater microbial persistence in sediments than in water over time was observed, along with differences in survival among microorganisms in relation to the fecal indicator bacteria E. coli and enterococci. Previous studies compared data among microbial groups in different environments. Conversely, fecal indicator bacteria, surrogates, and pathogenic microorganisms were assessed within the same water and sediment mesocosms in the present study during four seasons, better representing the agricultural aquatic environment. These data should be considered when agricultural microbial water quality criteria in fresh produce operations are being determined.

Swabbing the surface: critical factors in environmental monitoring and a path towards standardization and improvement.

Cross-contamination can be broadly defined as the transfer, direct or indirect, of microorganisms from a contaminated product to a non-contaminated product. Events that may result in cross-contamination include inadequate hygiene practices, contaminated equipment surfaces, contamination via food handling personnel, further product processing, or storage abuse All of these niches require consistent environmental surveillance systems to monitor microbial harborage sites to prevent foodborne illnesses via cross-contamination. Environmental surveillance is achieved through routine surface sampling of the food contact surfaces and surrounding areas. To better understand cross-contamination, the role of environmental surface transmission during outbreaks due to the presence and persistence of pathogenic microorganisms on various food contact surfaces must be investigated. However, studies on environmental sampling techniques are rarely performed in an actual food processing environment but rather under controlled variables within a laboratory-setting. Moreover, results and conclusions of studies differ because of the considerable variability across surface sampling tools due to individual operator dependency, low recovery rates, and low reproducibility. Information is also often lacking on environmental sampling tools used within a processing facility, the characterization of these tools, and the optimization of recovery of microorganisms for surface sampling. Thus, this review aims to: (1) discuss and compare factors impacting the recovery of microorganisms and the standardization of surface sampling methods for optimal recovery of microorganisms and (2) examine how research strategies could focus more towards the development of standard methodologies for surface sampling.

Virus-Bacteria Interactions: Implications for Prevention and Control of Human Enteric Viruses from Environment to Host.

Human enteric viruses, specifically human norovirus (hNoV), are the most common cause of foodborne illness boasting a wide range of transmission routes. These include person to person, contact with contaminated fomites, as well as ingestion of contaminated water and food. Because of this, the control and prevention of enteric viruses in food and other relevant environments have been a research focus over the past few decades. Interestingly, viruses as well as many other pathogens are often studied in isolation even though it is known that microorganisms do not occur in isolation but rather as part of complex microbial communities-both external from the host and within the host. Therefore, the overall goal of this review is to present the current evidence on virus-microbe interactions as these relate to the infectivity as well as the control and prevention of epidemiologically relevant foodborne viruses (such as hNoV) within our food systems. Therefore, this review is divided into in vivo, in situ, and in vitro implications of virus-microbe interactions through discussion of studies investigating the complex relationships between human enteric viruses and microbial cohabitants, specifically hNoV and bacteria.

Identification of Factors Affecting Fecal Pollution in Beaver Lake Reservoir.

Standard methods for the evaluation of recreational water quality rely on generic bacterial indicators such as . However, does not provide enough information to determine fecal source or public health risk. The stsudy objective was to determine factors influencing the presence of and host-specific markers (HSM) from upstream to downstream in Beaver Lake Reservoir (BLR). From February 2014 to September 2015, 420 base flow and rain event samples were collected from seven sites-two sites from streams (White River [WR] and War Eagle Creek) draining into BLR and five sites from within BLR. Each sample was analyzed for and by quantitative polymerase chain reaction for HSM related to human, bovine, and poultry. The data indicate that overall levels of were significantly greater in the WR and significantly lower at the most downstream sampling location in BLR. is more likely present during spring (adjusted odds ratio [aOR] = 1.86), at the WR sampling site (aOR = 3.39), or during a rain event (aOR = 2.73). Moreover, the HSM HumM2 is more likely present (aOR = 1.99) when is present. These same factors were associated with concentrations >126 most probable number 100 mL (aOR = 2.76-12.48), except the poultry marker CL was more likely associated (aOR = 3.81) than HumM2. This study revealed that both seasonal and locational factors are important variables for fecal pollution in BLR. Moreover, these same factors may apply to fecal pollution in manmade reservoirs within similar types of watersheds across the United States, as well as internationally.

Combined Application of Essential Oil Compounds and Bacteriophage to Inhibit Growth of Staphylococcus aureus In Vitro.

Staphylococcus aureus is considered an important human pathogen. This study aimed to investigate the combination of essential oil compounds (EOCs) and bacteriophage as alternative antimicrobials to control S. aureus in vitro. Here, four EOCs (alpha-pinene, 3-carene, (+)-limonene, (1S)-(-)-ß-pinene) were evaluated by disc diffusion assay (DDA) and growth inhibition assay (GIA) to determine inhibitory effects against five strains of S. aureus. Phage adsorption assays were performed with phage K up to 120 h at 6, 13, and 37 °C to determine lytic activity. Combinations of phage K and EOCs against S. aureus were also evaluated at 37 °C. Alpha-pinene exhibited significantly greater inhibition towards S. aureus strains when compared to other EOCs tested by the DDA. GIAs indicate that all S. aureus strains exhibited significantly reduced growth (P < 0.006) over a 48-h period when exposed to EOCs. Phage adsorption assays indicate that phage K has high lytic activity at 37 °C with at least a 1.5-log increase in the number of plaque-forming units (PFU) over 6 h when compared to 6 and 13 °C. S. aureus strains showed significantly reduced growth (P < 0.05) when treated with combined phage K and EOCs. Results from the combined effect of EOC and phage indicate that phage alone inhibits S. aureus in vitro at 37 °C as effectively as EOCs alone or in combination with phage with variability between strains. The results from DDA, GIA, and phage adsorption assays indicate that select EOCs and phage K can be used as antimicrobials against S. aureus in vitro with potential application in situ.

Interactions between Human Norovirus Surrogates and Acanthamoeba spp.

Human noroviruses (HuNoVs) are the most common cause of food-borne disease outbreaks, as well as virus-related waterborne disease outbreaks in the United States. Here, we hypothesize that common free-living amoebae (FLA)-ubiquitous in the environment, known to interact with pathogens, and frequently isolated from water and fresh produce-could potentially act as reservoirs of HuNoV and facilitate the environmental transmission of HuNoVs. To investigate FLA as reservoirs for HuNoV, the interactions between two Acanthamoeba species, A. castellanii and A. polyphaga, as well as two HuNoV surrogates, murine norovirus type 1 (MNV-1) and feline calicivirus (FCV), were evaluated. The results showed that after 1 h of amoeba-virus incubation at 25°C, 490 and 337 PFU of MNV-1/ml were recovered from A. castellanii and A. polyphaga, respectively, while only few or no FCVs were detected. In addition, prolonged interaction of MNV-1 with amoebae was investigated for a period of 8 days, and MNV-1 was demonstrated to remain stable at around 200 PFU/ml from day 2 to day 8 after virus inoculation in A. castellanii. Moreover, after a complete amoeba life cycle (i.e., encystment and excystment), infectious viruses could still be detected. To determine the location of virus associated with amoebae, immunofluorescence experiments were performed and showed MNV-1 transitioning from the amoeba surface to inside the amoeba over a 24-h period. These results are significant to the understanding of how HuNoVs may interact with other microorganisms in the environment in order to aid in its persistence and survival, as well as potential transmission in water and to vulnerable food products such as fresh produce.

Genes Indicative of Zoonotic and Swine Pathogens Are Persistent in Stream Water and Sediment following a Swine Manure Spill.

Manure spills into streams are relatively frequent, but no studies have characterized stream contamination with zoonotic and veterinary pathogens, or fecal chemicals, following a spill. We tested stream water and sediment over 25 days and downstream for 7.6 km for the following: fecal indicator bacteria (FIB), the fecal indicator chemicals cholesterol and coprostanol, 20 genes for zoonotic and swine-specific bacterial pathogens by presence/absence PCR for viable cells, one swine-specific Escherichia coli toxin gene (STII gene) by quantitative PCR (qPCR), and nine human and animal viruses by qPCR or reverse transcription-qPCR. Twelve days postspill, and 4.2 km downstream, water concentrations of FIB, cholesterol, and coprostanol were 1 to 2 orders of magnitude greater than those detected before, or above, the spill, and genes indicating viable zoonotic or swine-infectious Escherichia coli were detected in water or sediment. STII gene levels increased from undetectable before or above the spill to 10(5) copies/100 ml of water 12 days postspill. Thirteen of 14 water (8/9 sediment) samples had viable STII-carrying cells postspill. Eighteen days postspill, porcine adenovirus and teschovirus were detected 5.6 km downstream. FIB concentrations (per gram [wet weight]) in sediment were greater than in water, and sediment was a continuous reservoir of genes and chemicals postspill. Constituent concentrations were much lower, and detections less frequent, in a runoff event (200 days postspill) following manure application, although the swine-associated STII and stx2e genes were detected. Manure spills are an underappreciated pathway for livestock-derived contaminants to enter streams, with persistent environmental outcomes and the potential for human and veterinary health consequences.

Improving the Detection and Understanding of Infectious Human Norovirus in Food and Water Matrices: A Review of Methods and Emerging Models.

Human norovirus (HuNoV) is a leading global cause of viral gastroenteritis, contributing to numerous outbreaks and illnesses annually. However, conventional cell culture systems cannot support the cultivation of infectious HuNoV, making its detection and study in food and water matrices particularly challenging. Recent advancements in HuNoV research, including the emergence of models such as human intestinal enteroids (HIEs) and zebrafish larvae/embryo, have significantly enhanced our understanding of HuNoV pathogenesis. This review provides an overview of current methods employed for HuNoV detection in food and water, along with their associated limitations. Furthermore, it explores the potential applications of the HIE and zebrafish larvae/embryo models in detecting infectious HuNoV within food and water matrices. Finally, this review also highlights the need for further optimization and exploration of these models and detection methods to improve our understanding of HuNoV and its presence in different matrices, ultimately contributing to improved intervention strategies and public health outcomes.

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.

Hydroponic Nutrient Solution Temperature Impacts Tulane Virus Persistence over Time.

Controlled environment agriculture (CEA), or indoor agriculture, encompasses non-traditional farming methods that occur inside climate-controlled structures (e.g., greenhouses, warehouses, high tunnels) allowing for year-round production of fresh produce such as leaf lettuce. However, recent outbreaks and recalls associated with hydroponically grown lettuce contaminated with human pathogens have raised concerns. Few studies exist on the food safety risks during hydroponic cultivation of leaf lettuce; thus, it is important to identify contributing risk factors and potential mitigation strategies to prevent foodborne transmission via hydroponically grown produce. In this study, the concentration of infectious Tulane virus (TV), a human norovirus surrogate, in hydroponic nutrient solution at 15 °C, 25 °C, 30 °C, and 37 °C was determined over a duration of 21 days to mimic the time from seedling to mature lettuce. The mean log PFU reduction for TV was 0.86, 1.80, 2.87, and ≥ 3.77 log10 at 15 °C, 25 °C, 30 °C, and 37 °C, respectively, at the end of the 21-day period. Similarly, average decimal reduction values (D-values) of TV at 15 °C, 25 °C, 30 °C, and 37 °C were 48.0, 11.3, 8.57, and 7.02 days, respectively. This study aids in the (i) identification of possible food safety risks associated with hydroponic systems specifically related to nutrient solution temperature and (ii) generation of data to perform risk assessments within CEA leaf lettuce operations to inform risk management strategies for the reduction of foodborne outbreaks, fresh produce recalls, and economic losses.

Utilizing Zebrafish Embryos for Replication of Tulane Virus: A Human Norovirus Surrogate.

The zebrafish larvae/embryo model has been shown to support the replication of seven strains (G1.7[P7], GII.2[P16], GII.3[P16], GII.4[P4], GII.4[P16], GII.6[P7], and GII.17[P13]) of human norovirus (HuNoV). However, due to challenges in consistently obtaining HuNoV-positive stool samples from clinical sources, evaluating HuNoV surrogates in this model is highly valuable. This study assesses the potential of zebrafish embryos and larvae as a model for Tulane virus (TuV) replication. Three infection methods were examined: microinjection, immersion, and feeding. Droplet digital PCR was used to quantify viral RNA across all three infection methods. Microinjection of 3 nL of TuV into zebrafish embryos (< 6-h post-fertilization) resulted in significant replication, with viral RNA levels reaching 6.22 logs at 4-day post-infection. In contrast, the immersion method showed no replication after immersing 4-day post-fertilization (dpf) larvae in TuV suspension for 6 h. Similarly, no replication was observed with the feeding method, where Paramecium caudatum loaded with TuV were fed to 4 dpf larvae. The findings indicate that the zebrafish embryo model supports TuV replication through the microinjection method, suggesting that TuV may serve as a useful surrogate for studying HuNoV pathogenesis. Additionally, TuV can be utilized in place of HuNoV in method optimization studies using the zebrafish embryo model, circumventing the limited availability of HuNoV.

A comprehensive examination of microbial hazards and risks during indoor soilless leafy green production.

Produce grown under controlled environment agriculture (CEA) is often assumed to have a reduced risk of pathogen contamination due to the low chance of exposure to outdoor contaminant factors. However, the 2021 outbreak and numerous recalls of CEA-grown lettuce and microgreens demonstrate the possibility of pathogen introduction during indoor production when there is a failure in the implementation of food safety management systems. Indoor production of commercial leafy greens, such as lettuce and microgreens, is performed across a range of protective structures from primitive household setups to advanced and partially automatized growing systems. Indoor production systems include hydroponic, aquaponic, and aeroponic configurations. Hydroponic systems such as deep water culture and nutrient film technique comprised of various engineering designs represent the main system types used by growers. Depending on the type of leafy green, the soilless substrate, and system selection, risk of microbial contamination will vary during indoor production. In this literature review, science-based pathogen contamination risks and mitigation strategies for indoor production of microgreens and more mature leafy greens are discussed during both pre-harvest and post-harvest stages of production.

Functional dependence as a contributing factor for patient hand contamination by multi-drug resistant organisms (MDROs) in acute care.

OBJECTIVE: Patient hands' contribution to disease transmission in healthcare settings is an important, understudied topic. We assessed correlation between patient functional dependence and hand contamination with multi-drug resistant organisms (MDROs) in acute-care settings. DESIGN, SETTING, AND PATIENTS: Secondary, cross-sectional analyses of 399 general medicine patients enrolled in two tertiary-care hospitals over a six-month period. Our predictor was patient functional status evaluated using Katz Activities of Daily Living scale, scored as follows: functionally independent (scored 0), moderately dependent (score 1-3), and severely dependent (score of 4 or more). Our outcome was patient hand contamination with MDROs, including methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus, and resistant gram-negative bacteria. RESULTS: Of 399 patients, 298 were functionally independent, 45 were moderately dependent, and 56 were severely dependent. Odds of MDRO hand contamination were 2.63 (95% CI, 1.21-5.72) times higher in the severely dependent category compared to the independent category. Patients with feeding dependence had the highest odds of hand contamination with MDROs (OR 4.76, 95% CI, 1.54-14.71), followed by continence, dressing, and toileting. In addition to patient colonization, environmental contamination with MRSA was associated with patient function, with odds 2.60 (95% CI, 1.16-5.82) times higher in severely dependent patients. CONCLUSIONS: Patients with severe functional dependence are more likely to harbor MDROs on their hands and less likely to be able to cleanse them independently. Functionally dependent patients have high room contamination with MDROs. Patient hand hygiene interventions in the hospital should target this high-risk group.

Does high school health education class satisfaction influence student perceptions of public health and pandemic response?

BACKGROUND: The COVID-19 pandemic significantly affected high school students. Little is known about the mediators of student perceptions of infection prevention and public health entities. We piloted a survey to evaluate the relationship between student perceptions of COVID-19 topics and satisfaction with their most recent health class. METHODS: Students from one private high school in southeast Michigan completed a survey in early 2022. The primary outcomes were 4 domains: vaccination knowledge, intervention effectiveness, intervention impact, and willingness to readopt an intervention. We assessed the associations between health class satisfaction and these outcomes using multiple linear regression. RESULTS: One-hundred ninety students reported their health class satisfaction and were eligible for analysis. Students reported high confidence in vaccines (93%) but limited knowledge of COVID-19 vaccination (45%). Students perceived COVID-19 interventions as highly effective (range, 72% [hand hygiene]-93% [vaccination]) and reported a willingness to readopt them (range, 73% [stay-at-home orders]-96% [vaccination]). Health class satisfaction (54%) was positively associated with composite scores on vaccination knowledge and intervention effectiveness. DISCUSSION: Assessing students' intrapandemic perspectives on infection prevention illuminated areas of strength (ie, intervention confidence) and areas for improvement (ie, intervention knowledge and institutional confidence). CONCLUSIONS: Students reported favorable perceptions regarding common infection prevention interventions. Future work should investigate the role of educational satisfaction in mediating confidence in public health interventions and institutions.

Post-acute sequelae of SARS-CoV-2 (PASC) in nursing home residents: A retrospective cohort study.

BACKGROUND: Post-acute sequelae of SARS-CoV-2 (PASC) describes a syndrome of physical and cognitive decline that persists after acute symptoms of infection resolve. Few studies have explored PASC among nursing home (NH) residents. METHODS: A retrospective cohort study was conducted at two NHs in Michigan. COVID-positive patients were identified from March 21, 2020 to October 26, 2021. The comparison group were patients who lived at the same NH but who were never infected during the study period. Minimum Data Set was used to examine trajectories of functional dependence (Activity of Daily Living [ADL] composite score) and cognitive function (Brief Interview for Mental Status [BIMS]). Linear mixed-effects models were constructed to estimate short-term change in function and cognition immediately following diagnosis and over time for an additional 12 months, compared to pre-COVID and non-COVID trajectories and adjusting for sex, age, and dementia status. RESULTS: We identified 171 residents (90 COVID-19 positive, 81 non-COVID) with 719 observations for our analyses. Cohort characteristics included: 108 (63%) ≥ 80 yrs.; 121 (71%) female; 160 (94%) non-Hispanic white; median of 3 comorbidities (IQR 2-4), with no significant differences in characteristics between groups. COVID-19 infection affected the trajectory of ADL recovery for the first 9 months following infection, characterized by an immediate post-infection decrease in functional status post-infection (-0.60 points, p = 0.002) followed by improvement toward the expected functional trajectory sans infection (0.04 points per month following infection, p = 0.271). CONCLUSIONS: NH residents experienced a significant functional decline that persisted for 9 months following acute infection. Further research is needed to determine whether increased rehabilitation services after COVID-19 may help mitigate this decline.

Machine Learning and Imputation to Characterize Human Norovirus Genotype Susceptibility to Sodium Hypochlorite.

Human norovirus (HuNoV) is the leading cause of foodborne illness in the developed world and a major contributor to gastroenteritis globally. Its low infectious dose and environmental persistence necessitate effective disinfection protocols. Sodium hypochlorite (NaOCl) bleach is a widely used disinfectant for controlling HuNoV transmission via contaminated fomites. This study aimed to evaluate the susceptibility of HuNoV genotypes (n = 11) from genogroups I, II, and IV to NaOCl in suspension. HuNoV was incubated for 1 and 5 min in diethyl pyrocarbonate (DEPC) treated water containing 50 ppm, 100 ppm, or 150 ppm NaOCl, buffered to maintain a pH between 7.0 and 7.5. Neutralization was achieved by a tenfold dilution into 100% fetal bovine serum. RNase pre-treatment followed by RT-qPCR was used to distinguish between infectious and non-infectious HuNoV. Statistical methods, including imputation, machine learning, and generalized linear models, were applied to process and analyze the data. Results showed that NaOCl reduced viral loads across all genotypes, though efficacy varied. Genotypes GI.1, GII.4 New Orleans, and GII.4 Sydney were the least susceptible, while GII.6 and GII.13 were the most susceptible. All NaOCl concentrations above 0 ppm were statistically indistinguishable, and exposure duration did not significantly affect HuNoV reduction, suggesting rapid inactivation at effective concentrations. For instance, some genotypes were completely inactivated within 1 min, rendering extended exposure unnecessary, while other genotypes maintained the initial concentration at both 1 and 5 min, indicating a need for longer contact times. These findings underscore the critical role of HuNoV genotype selection in testing disinfection protocols and optimizing NaOCl concentrations. Understanding HuNoV susceptibility to NaOCl bleach informs better disinfection strategies, aiding public health and food safety authorities in reducing HuNoV transmission and outbreaks.