Superhydrophobic coatings reduce human bacterial foodborne pathogen attachment to woods used in fresh produce harvest and postharvest packing.

Wood is reportedly more difficult to maintain in hygienic condition versus other food contact materials, yet its use in produce packing and retail warrants efforts to reduce the risk of microbial pathogen contamination and attachment. This study characterized antifouling capabilities of fluorinated silanes applied to wood used in fresh edible produce handling to render the wood superhydrophobic and less supportive of bacterial pathogen attachment. Pine and oak cubic coupon surfaces were treated with 1% (w/w) silane or left untreated. Treated and untreated coupons were inoculated with Salmonella enterica or Listeria monocytogenes and held to facilitate pathogen attachment for 1, 4, or 8 h. Silane treatment of wood produced significant reductions in the proportions of strongly attaching cells for both pathogens versus loosely attaching cells (P < 0.01). Salmonella attachment demonstrated a dependency on wood treatment; silane-treated wood supported a lower fraction of strongly adhering cells (1.87 ± 1.24 log CFU/cm2) versus untreated wood (3.72 ± 0.67 log CFU/cm2). L. monocytogenes demonstrated significant declines in strongly attaching cells during extended exposure to silane-treated wood, from 7.59 ± 0.14 to 5.27 ± 0.68 log CFU/cm2 over 8 h post-inoculation. Microscopic analysis demonstrated silane treatment increased the surface roughness of both woods, leading to superhydrophobic conditions on wood surfaces, consequently decreasing strong attachment of pathogenic bacteria.

Southern Region Produce Safety Alliance Grower Training: Using Pre- and Post-Training Knowledge Assessments to Understand Training Effectiveness.

The Produce Safety Alliance (PSA) grower training was introduced in 2016 as the standardized curriculum to meet the training requirements of the Food and Drug Administration's (FDA) Food Safety Modernization Act's (FSMA) Produce Safety Rule (PSR). The PSR states that at least one supervisor or responsible party from each farm must have successfully completed this food safety training or one equivalent to the standardized curriculum, as recognized by the FDA. This study evaluated the effectiveness of PSA trainings conducted between 2017 and 2019 in the Southern United States by the Southern Regional Center for Food Safety Training, Outreach, and Technical Assistance by analyzing pre- and posttest assessments. Effectiveness was based on a 25-question knowledge assessment administered to participants before (n = 2494) and after (n = 2460) each training. The knowledge assessment indicated the overall effectiveness of the training, with average scores increasing significantly from pretest (15.9/25, 63.4%) to posttest (20.3/25, 81.3%) (P < 0.001). The greatest knowledge gains were seen in the Postharvest Handling and Sanitation, How to Develop a Farm Food Safety Plan, and Agricultural Water modules. Notably, these modules had lower posttest scores compared to the other modules, indicating that the amount of knowledge gained did not necessarily correspond with a sufficient understanding of the material. To ensure that participants understand all aspects of the PSR and best practices to minimize food safety risks, additional or advanced trainings may be needed. Additionally, the current testing instrument (pre-/posttest) used for PSA grower training, while validated, may not be optimal, thus alternative methods to assess the training effectiveness are likely needed.

Edible nano-encapsulated cinnamon essential oil hybrid wax coatings for enhancing apple safety against food borne pathogens.

Post-harvest losses of fruits due to decay and concerns regarding microbial food safety are significant within the produce processing industry. Additionally, maintaining the quality of exported commodities to distant countries continues to pose a challenge. To address these issues, the application of bioactive compounds, such as essential oils, has gained recognition as a means to extend shelf life by acting as antimicrobials. Herein, we have undertaken an innovative approach by nano-encapsulating cinnamon-bark essential oil using whey protein concentrate and imbibing nano-encapsulates into food-grade wax commonly applied on produce surfaces. We have comprehensively examined the physical, chemical, and antimicrobial properties of this hybrid wax to evaluate its efficacy in combatting the various foodborne pathogens that frequently trouble producers and handlers in the post-harvest processing industry. The coatings as applied demonstrated a static contact angle of 85 ± 1.6°, and advancing and receding contact angles of 90 ± 1.1° and 53.0 ± 1.6°, respectively, resembling the wetting properties of natural waxes on apples. Nanoencapsulation significantly delayed the release of essential oil, increasing the half-life by 61 h compared to its unencapsulated counterparts. This delay correlated with statistically significant reductions (p = 0.05) in bacterial populations providing both immediate and delayed (up to 72 h) antibacterial effects as well as expanded fungal growth inhibition zones compared to existing wax technologies, demonstrating promising applicability for high-quality fruit storage and export. The utilization of this advanced produce wax coating technology offers considerable potential for bolstering food safety and providing enhanced protection against bacteria and fungi for produce commodities.

Reduction of Bacterial Enteric Pathogens and Hygiene Indicator Bacteria on Tomato Skin Surfaces by a Polymeric Nanoparticle-Loaded Plant-Derived Antimicrobial.

This study determined Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium survival on tomato skins as a function of sanitization treatment, under three differing contamination and sanitization scenarios. Sanitizing treatments consisted of the plant-derived antimicrobial (PDA) geraniol (0.5 wt.%) emulsified in the polymeric surfactant Pluronic F-127 (GNP), 0.5 wt.% unencapsulated geraniol (UG), 200 mg/L hypochlorous acid at pH 7.0 (HOCl), and a sterile distilled water wash (CON). Experimental contamination and sanitization scenarios tested were: (1) pathogen inoculation preceded by treatment; (2) the pathogen was inoculated onto samples twice with a sanitizing treatment applied in between inoculations; and (3) pathogen inoculation followed by sanitizing treatment. Reductions in counts of surviving pathogens were dependent on the sanitizing treatment, the storage period, or the interaction of these independent/main effects. GNP treatment yielded the greatest reductions in pathogen counts on tomato skins; pathogen survivor counts following GNP treatment were consistently statistically lower than those achieved by HOCl or UG treatments (p < 0.05). GNP treatment provided greatest pathogen reduction under differing conditions of pre- and/or post-harvest cross-contamination, and reduced hygiene-indicating microbes the most of all treatments on non-inoculated samples. Encapsulated geraniol can reduce the risk of pathogen transmission on tomato fruit, reducing food safety hazard risks for tomato consumers.

Complete Whole Genome Sequences of Escherichia coli Surrogate Strains and Comparison of Sequence Methods with Application to the Food Industry.

In 2013, the U.S. Department of Agriculture Food Safety and Inspection Service (USDA-FSIS) began transitioning to whole genome sequencing (WGS) for foodborne disease outbreak- and recall-associated isolate identification of select bacterial species. While WGS offers greater precision, certain hurdles must be overcome before widespread application within the food industry is plausible. Challenges include diversity of sequencing platform outputs and lack of standardized bioinformatics workflows for data analyses. We sequenced DNA from USDA-FSIS approved, non-pathogenic E. coli surrogates and a derivative group of rifampicin-resistant mutants (rifR) via both Oxford Nanopore MinION and Illumina MiSeq platforms to generate and annotate complete genomes. Genome sequences from each clone were assembled separately so long-read, short-read, and combined sequence assemblies could be directly compared. The combined sequence data approach provides more accurate completed genomes. The genomes from these isolates were verified to lack functional key E. coli elements commonly associated with pathogenesis. Genetic alterations known to confer rifR were also identified. As the food industry adopts WGS within its food safety programs, these data provide completed genomes for commonly used surrogate strains, with a direct comparison of sequence platforms and assembly strategies relevant to research/testing workflows applicable for both processors and regulators.

Validating Thermal Lethality to Salmonella enterica in Chicken Blood by Simulated Commercial Rendering.

The U.S. rendering industry produces materials for use in further processed animal foods and feeds and is required to scientifically validate food safety hazard control. This study aimed to provide lethality validation for Salmonella enterica during simulated commercial rendering of whole chicken blood. Chicken blood was inoculated with a blend of multiple serovars of the pathogen (S. Heidelberg, Typhimurium, Senftenberg) and subjected to heating at 82.2, 87.8, or 93.3 °C; surviving cells were enumerated incrementally up to 5.0 min. Survivor data were modeled using the GInaFiT 1.7 freeware package. D-values and t7D (time to a 7.0 log10-cycle inactivation) values were generated from best-fit model parameters. Predictive modeling analysis revealed that the survival curves of Salmonella possessed log-linear components but also possessed shoulder and/or tail components. Mean D-values declined from 0.61 to 0.12 min as heating temperature was raised from 82.2 to 93.3 °F, respectively, differing by heating temperature (p = 0.023). t7D values differed significantly by heating temperature (p = 0.001), as was also the case for shoulder length (SL) (p = <0.0001), where, at lower temperatures, a shoulder was observed versus heating at 93.3 °F. These data aid scientific validation of Salmonella enterica inactivation during thermal rendering of poultry blood for use in further processed animal foods.

Application of Surfactant Micelle-Entrapped Eugenol for Prevention of Growth of the Shiga Toxin-Producing Escherichia coli in Ground Beef.

Beef safety may be compromised by O157 and non-O157 Shiga toxin-producing Escherichia coli (STEC) contamination. The capacity of surfactant micelles loaded with the plant-derived antimicrobial eugenol to reduce STEC on beef trimmings that were later ground and refrigerated for five days at 5 ± 1 °C was tested to determine their utility for beef safety protection. STEC-inoculated trimmings were treated with free eugenol, micelle-encapsulated eugenol, 2% lactic acid (55 °C), sterile distilled water (25 °C), or left untreated (control). Following treatment, trimmings were coarse-ground and stored aerobically at 5 ± 1 °C. Ground beef was then sampled for STEC immediately post-grinding, and again at three and five days of storage. STEC minimum inhibitory concentrations (MICs) in liquid medium for free eugenol and 1% sodium dodecyl sulfate (SDS)-loaded micelles were 0.5% and 0.125%, respectively. STEC numbers on beef trimmings treated by sterile water (6.5 log10 CFU/g), free eugenol (6.5 log10 CFU/g), micelle-loaded eugenol (6.4 log10 CFU/g), and lactic acid (6.4 log10 CFU/g) did not differ compared to untreated controls (6.6 log10 CFU/g) (p = 0.982). Conversely, STEC were significantly reduced by refrigerated storage (0.2 and 0.3 log10 CFU/g at three and five days of storage, respectively) (p = 0.014). Antimicrobial treatments did not significantly decontaminate ground beef, indicating their low utility for beef safety protection.

Investigation into Formation of Lipid Hydroperoxides from Membrane Lipids in Escherichia coli O157:H7 following Exposure to Hot Water.

Although studies have shown antimicrobial treatments consisting of hot water sprays alone or paired with lactic acid rinses are effective for reducing Escherichia coli O157:H7 loads on beef carcass surfaces, the mechanisms by which these interventions inactivate bacterial pathogens are still poorly understood. It was hypothesized that E. coli O157:H7 exposure to hot water in vitro at rising temperatures for longer time periods would result in increasing deterioration of bacterial outer membrane lipids, sensitizing the pathogen to subsequent lactic acid application. Cocktails of E. coli O157:H7 strains were subjected to hot water at 25 (control) 65, 75, or 85 °C incrementally up to 60 s, after which surviving cells were enumerated by plating. Formation of lipid hydroperoxides from bacterial membranes and cytoplasmic accumulation of L-lactic acid was quantified spectrophotometrically. Inactivation of E. coli O157:H7 proceeded in a hot water exposure duration- and temperature-dependent manner, with populations being reduced to nondetectable numbers following heating of cells in 85 °C water for 30 and 60 s (P < 0.05). Lipid hydroperoxide formation was not observed to be dependent upon increasing water temperature or exposure period. The data suggest that hot water application prior to organic acid application may function to increase the sensitivity of E. coli O157:H7 cells by degrading membrane lipids.

Inhibition of Listeria monocytogenes by food antimicrobials applied singly and in combination.

Combining food antimicrobials can enhance inhibition of Listeria monocytogenes in ready-to-eat (RTE) meats. A broth dilution assay was used to compare the inhibition of L. monocytogenes resulting from exposure to nisin, acidic calcium sulfate, ε-poly-L-lysine, and lauric arginate ester applied singly and in combination. Minimum inhibitory concentrations (MICs) were the lowest concentrations of single antimicrobials producing inhibition following 24 h incubation at 35 °C. Minimum bactericidal concentrations (MBCs) were the lowest concentrations that decreased populations by ≥3.0 log(10) CFU/mL. Combinations of nisin with acidic calcium sulfate, nisin with lauric arginate ester, and ɛ-poly-L-lysine with acidic calcium sulfate were prepared using a checkerboard assay to determine optimal inhibitory combinations (OICs). Fractional inhibitory concentrations (FICs) were calculated from OICs and were used to create FIC indices (FIC(I)s) and isobolograms to classify combinations as synergistic (FIC(I) < 1.00), additive/indifferent (FIC(I)= 1.00), or antagonistic (FIC(I) > 1.00). MIC values for nisin ranged from 3.13 to 6.25 µg/g with MBC values at 6.25 µg/g for all strains except for Natl. Animal Disease Center (NADC) 2045. MIC values for ε-poly-L-lysine ranged from 6.25 to 12.50 µg/g with MBCs from 12.50 to 25.00 µg/g. Lauric arginate ester at 12.50 µg/g was the MIC and MBC for all strains; 12.50 mL/L was the MIC and MBC for acidic calcium sulfate. Combining nisin with acidic calcium sulfate synergistically inhibited L. monocytogenes; nisin with lauric arginate ester produced additive-type inhibition, while ε-poly-L-lysine with acidic calcium sulfate produced antagonistic-type inhibition. Applying nisin along with acidic calcium sulfate should be further investigated for efficacy on RTE meat surfaces.

The use of reduced healing times on ITI implants with a sandblasted and acid-etched (SLA) surface: early results from clinical trials on ITI SLA implants.

ITI dental implants are available with two bone-anchoring surfaces, a titanium plasma-sprayed (TPS) surface, and a recently introduced sandblasted and acid-etched (SLA) surface. Cell culture and animal tests demonstrate that the SLA surface stimulates bone cell differentiation and protein production, has large amounts of bone-to-implant contact, and results in large removal torque values in functional testing of the bone contact. As a result of these studies, a prospective human clinical trial was initiated to determine whether the 4.1 mm diameter SLA ITI solid screw implants could be predictably and safely restored as early as six weeks after implant placement surgery. The protocol restricted the use of the reduced healing time to a) healthy patients with sufficient bone volume to surround the implant, and b) those patients who had good bone quality (classes I-III) at the implant recipient site. Patients with poorer bone quality (class IV) did not have restorations until 12 weeks after implant placement. The clinical trial is an ongoing multicenter trial, with six centers in four countries, and with follow-up over five years. The primary outcome variable was abutment placement with a 35 Ncm force, with no countertorque and no pain or rotation of the implant. A secondary outcome was implant success, as defined by no mobility, no persistent pain or infection, and no peri-implant radiolucency. To date, 110 patients with 326 implants have completed the one-year post-loading recall visit, while 47 patients with 138 implants have completed the two-year recall. Three implants were lost prior to abutment connection. Prosthetic restoration was commenced after shortened healing times on 307 implants. The success rate for these implants, as judged by abutment placement, was 99.3% (with an average healing time of 49 days). Life table analyses demonstrated an implant success rate of 99.1%, both for 329 implants at one year and for 138 implants at two years. In the 24-month period after restoration, no implant losses were reported for the 138 implants. These results demonstrate that, under defined conditions, solid screw ITI implants with an SLA endosseous surface can be restored after approximately six weeks of healing with a high predictability of success, defined by abutment placement at 35 Ncm without countertorque, and with subsequent implant success rates of greater than 99% two years after restoration.