Category Archives: Technology

Research – Evaluating a Combined Method of UV and Washing for Sanitizing Blueberries, Tomatoes, Strawberries, Baby Spinach, and Lettuce

Journal of Food Protection

ABSTRACT

We assessed a fresh produce decontamination system using a combined method of UV and washing (water-assisted UV [WUV]) in different scales. The system used tap water to wash fresh produce while exposing it to UV light. First, the reduction of Salmonella in tap water under UV treatment (1 to 1,740 mJ/cm2) was determined. Increasing the UV dose significantly (P < 0.05) increased the Salmonella reduction in wash water, and UV intensity of more than 2 mW/cm2 could reduce Salmonella in tap water to below 1 CFU/mL given enough processing time (more than 1 min; UV dose of 120 mJ/cm2). Then, the decontamination effectiveness of a small WUV system was tested on blueberries (50 g). Blueberries were spot or dip inoculated with a Salmonella cocktail and treated by the small WUV system (200 mL of water). In general, WUV treatments achieved significantly better Salmonella inactivation than tap water wash; tap water wash (10 min) and 2 mW/cm2 WUV treatment (with a UV dose of 1,200 mJ/cm2) reduced populations of spot-inoculated Salmonella on blueberries by 2.44 and 5.45 log, respectively. Compared with spot-inoculated Salmonella on blueberries, dip-inoculated Salmonella was more difficult to be inactivated by WUV treatments. Then, the decontamination effectiveness of WUV treatments was tested on blueberries (170 g), tomatoes (290 g), strawberries (170 g), baby spinach (60 g), and lettuce (60 g) using a larger WUV system. In general, 10 min of 29 mW/cm2 WUV treatment (a high UV dose of 17,400 mJ/cm2) resulted in significantly better Salmonella inactivation than tap water wash (for 10 min) regardless the inoculation method, agreeing with the results of the small-scale study. For both spot- and dip-inoculated lettuce, no significant difference (P > 0.05) in Salmonella inactivation by WUV treatments was observed when the quantity of lettuce increased from 50 to 100 g.

HIGHLIGHTS
  • Water-assisted UV showed ≤3 log more Salmonella inactivation than tap water wash.

  • Water-assisted UV showed the highest Salmonella reduction on blueberries and tomatoes.

  • More spot-inoculated Salmonella was killed than dip-inoculated Salmonella by up to 3 log.

  • Water-assisted UV of 17.4 J/cm2 could reduce Salmonella in water to less than 1 CFU/mL.

Research – Interaction and inactivation of Listeria and Lactobacillus cells in single and mixed species biofilms exposed to different disinfectants

Wiley Online

Abstract

Listeria spp. are ubiquitously found in both the natural and the food processing environment, of which Listeria monocytogenes is of an important health risk. Here, we report on the formation of single and mixed species biofilms of L. monocytogenes/Listeria innocua and Lactobacillus plantarum strains in 24‐well polystyrene microtiter plates and on the inactivation of 24‐hr and 72‐hr biofilms using quaternary ammonium compound‐, tertiary alkyl amine‐, and chlorine‐based disinfectants. Fluorescent in situ hybridization (FISH) and LIVE/DEAD BacLight staining were applied for 72‐hr L. innocuaL. plantarum mixed biofilms in the LabTek system for the species identification and the reaction of biofilm cells to disinfectants, respectively. L. monocytogenes/L. innocua were more resistant to disinfectants in 72‐hr than in 24‐hr biofilms, whereas L. plantarum strains did not show any significant differences between 72‐hr and 24‐hr biofilms. Furthermore, L. innocua when grown with L. plantarum was more resistant to all disinfection treatments, indicating a protective effect from lactobacilli in the mixed species biofilm. The biofilm formation and reaction to disinfectants, microscopically verified using fluorescence in situ hybridization and LIVE/DEAD staining, showed that L. innocua and L. plantarum form a dense mixed biofilm and also suggested the shielding effect of L. plantarum on L. innocua in the mixed species biofilm.

Research – Antibiofilm Efficacy of Peptide 1018 against Listeria monocytogenes and Shiga Toxigenic Escherichia coli on Equipment Surfaces

Journal of Food Protection

ABSTRACT

Listeria monocytogenes and Shiga toxigenic Escherichia coli (STEC) are important foodborne bacterial pathogens that can form biofilms on equipment surfaces at food processing facilities. Pathogens in biofilms are resistant to conventional antimicrobials and require higher antimicrobial concentrations to be inactivated. In this study, the efficacy of a synthetic innate defense regulator peptide 1018 (peptide 1018) for inactivating L. monocytogenes and STEC (O26, O111, O145, O157) biofilms on stainless steel and polycarbonate surfaces was investigated. Stainless steel and polycarbonate coupons (12 mm in diameter) were used in a Centers for Disease Control and Prevention biofilm reactor containing 400 mL of 10% tryptic soy broth (TSB) that had been inoculated with an individual strain of L. monocytogenes or STEC to obtain 6 log CFU/mL populations. The reactor was set with a constant flow rate at 50 mL/h of 10% TSB for 48 h. After 48 h, coupons were treated with peptide 1018 at 0, 10, 20, or 50 μg/mL in phosphate buffer saline (PBS) for 24 h. Surviving bacterial populations were determined by scraping off the coupons and spiral plating on selective media. Significantly higher levels of pathogens in biofilms formed by certain bacterial strains, including L. monocytogenes F6854, E. coli O157:H7 RM4407 and NADC5713, and non-O157 E. coli NADC3629, were recovered on polycarbonate surfaces than on stainless steel. Antibiofilm efficacy of peptide 1018 against pathogens was concentration-dependent and varied with the type of pathogen and material surfaces. Peptide 1018 at 50 μg/mL significantly inactivated all tested bacterial biofilms on both surfaces compared with the PBS control (P < 0.05). L. monocytogenes was the bacterium most sensitive to peptide 1018; on stainless steel surfaces treated with 50 μg/mL peptide 1018, there was a 3.7- to 4.6-log CFU/cm2 reduction in Listeria populations compared with a 1.0- to 3.5-log CFU/cm2 reduction of STEC. Results suggest that peptide 1018 may be used to inactivate L. monocytogenes and STEC biofilms on equipment surfaces.

HIGHLIGHTS
  • Bacteria attach at higher levels on polycarbonate surfaces than on stainless steel.

  • L. monocytogenes is more sensitive than STEC to peptide 1018.

  • Peptide 1018 can be used to inactivate biofilms on equipment surfaces.

Research – Anti‐listeria activity and shelf life extension effects of Lactobacillus along with garlic extract in ground beef

Wiley Online

The current study investigates the effect of Lactobacillus reuteri and Lactobacillus plantarum combined with water extract of garlic on microbial growth, chemical changes, and sensory attributes in ground beef samples at refrigeration condition (+4°C) up to 12 days of storage. in vitro study revealed that garlic extract combined with L. reuteri or L. plantarum caused 2.13 and 2.57 log reduction in the Listeria monocytogenes count, respectively. Combination of L. plantarum and 1% garlic extract significantly (p < .05) reduced aerobic mesophilic bacteria (1.64 log cycle) and L. monocytogenes (1.44 log cycle) counts in ground beef. Lipid oxidation was also significantly (p < .05) lower in samples treated with L. plantarum plus garlic extract (1%). Furthermore, higher sensory scores were received by samples treated with Lactobacillus plus garlic extract. In conclusion, the combination of L. plantarum and garlic extract was found to be suitable to use in ground beef by controlling the L. monocytogenes growth and increasing its shelf life.

Practical Applications

Garlic extract not only has an antimicrobial activity but also has a stimulatory effect on the Lactobacillus spp. growth. On the other hand, some Lactobacillus strains can inhibit pathogenic bacteria. Then, the combination of Lactobacillus and garlic extract may be used to produce new bio‐preserved and functional meat products. The current study indicated the potential of Lactobacillus combined with garlic extract to control microbial and chemical changes in ground beef. The combination of Lactobacillus plantarum and garlic extract significantly (p < .05) reduced Listeria monocytogenes counts and lipid oxidation rates and improved the sensory scores in ground beef.

Research – Fate of Spoilage and Pathogenic Microorganisms in Acidified Cold-Filled Hot Pepper Sauces

Journal of Food Protection

ABSTRACT

Consumption of spicy foods and hot sauces is currently a popular trend worldwide. Shelf-stable acidified sauces are commonly hot-filled to ensure commercial sterility, but cold-fill-hold processes might also be suitable if microbial safety and stability are ensured. For this study, model acidified hot pepper sauces were developed and characterized. The effects of sauce pH and of two different organic acids on the survival of Pichia manshurica and Lactobacillus curvatus isolated from contaminated commercial hot sauces and on pathogenic Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes were assessed. Full factorial designs with three levels for pH (3.2, 3.5, and 3.9) and two for organic acid (citric and acetic) were used to determine the effects of these factors and their interactions on the survival of the microorganisms. Commercially sterile sauces were independently inoculated and kept at ambient temperature. Microbial counts were determined at different sampling times, depending on the treatment evaluated. Sauces acidified to pH 3.2 with citric or acetic acid were inoculated with cocktails of five strains or serotypes of the three pertinent pathogens, and inactivation curves were determined. Trials were performed in triplicate. A greater than 5-log reduction of P. manshurica and L. curvatus was achieved in less than 6 h in sauces adjusted to pH 3.2 with acetic acid. Greater than 5-log reductions of pathogenic bacteria were achieved 0.5 h after inoculation in sauces acidified to pH 3.2 with acetic acid. In contrast, at least 48 h was required to guarantee the same inactivation for the most tolerant pathogen when citric acid was used. Thus, a cold-fill-hold process may be a suitable alternative for acidified hot pepper sauces. Based on survival of the microorganisms evaluated in this study, microbial safety and stability can be achieved by adjusting the pH to 3.2 or less by the addition of acetic acid.

HIGHLIGHTS
  • pH and acidifier influence safety and stability of cold-filled acidified sauces.

  • Safe and stable cold-filled sauces were obtained at pH 3.2 with acetic acid.

  • Results help establish science-based conditions for cold-filled products.

Research – Validation of a High-Throughput Sausage Casing Model for the Assessment of Bacterial Inactivation Affected by Salt Concentration, pH, and Temperature

Journal of Food Protection

ABSTRACT

Previous studies have shown the efficacy of high concentrations of salt as the main preservative against vegetative bacteria present on natural sausage casings. These studies were limited in the number of variables and the interactions between these variables that were assessed. To remedy this situation, a MicroCasing high-throughput model was developed and validated to study the inactivation kinetics of various combinations of parameters (salt concentration, pH, and temperature) on eight bacterial isolates of Salmonella enterica, Staphylococcus aureus, Escherichia coli, and Listeria monocytogenes over a prolonged period. A Weibullian power model was the best fit to show the trends in sensitivity of each bacterial isolate to salt, pH, and temperature over time. The inactivation kinetics generated with this novel approach could serve as a predictive model for the required salting period for casings. The actual bacterial contamination of the product can vary with the respective production step during processing from animal intestine into sausage casings (initial level, ∼105 CFU/g; level after salting, <102 CFU/g). Subsequent selection and grading of these casings will require complete removal of all salt, and upon completion of this production step, the casings will be resalted. By determining the actual contamination level before the salting process, the minimum storage period in salt can be calculated and potentially optimized by adjusting the pH and temperature. As a result, a standard holding period of at least 30 days may no longer be necessary to produce salted natural casings in accordance with validated quality and food safety criteria.

HIGHLIGHTS
  • A new model system was developed for analysis of bacterial inactivation kinetics in foods.

  • The novel model allows determination of product-specific bacterial inactivation over time.

  • Effects of time, temperature, pH, and salt on casing preservation can be clarified with the model.

  • Prediction of inactivation parameters allows casing production to meet HACCP criteria.

Research – Microbiological Testing Results of Boneless and Ground Beef Purchased for the U.S. National School Lunch Program, School Years 2015 to 2018

Journal of Food Protection

ABSTRACT

The Agricultural Marketing Service (AMS) purchases beef for the National School Lunch Program and other federal nutrition assistance programs. For beef that will be delivered to food service facilities raw, each ca. 900-kg lot of boneless beef raw material and each ca. 4,500-kg sublot of resultant ground beef is tested for standard plate count (SPC) organisms, coliforms, Escherichia coli, Salmonella, and E. coli O157:H7. In addition, 1 of every 10 lots of boneless beef, randomly selected, is tested for E. coli O26, O45, O103, O111, O121, and O145. For beef that will be cooked using a validated lethality step at a federally inspected establishment before delivery, each lot of boneless beef and each sublot of ground beef is tested for SPC organisms, coliforms, and E. coli only. Any lot or sublot exceeding predefined critical limits (CLs) of 100,000 CFU g−1 for SPC organisms, 1,000 CFU g−1 for coliforms, or 500 CFU g−1 for E. coli or for beef containing Salmonella or any of previously mentioned E. coli serotypes is rejected for purchase. For school years 2015 through 2018 (July 2014 through June 2018), 220,497,254 kg of boneless beef and 189,347,318 kg of ground beef were produced for AMS. For boneless beef, 133 (0.06%), 164 (0.07%), and 106 (0.04%) of 240,488 lots exceeded CLs for SPC organisms, coliforms, and E. coli, respectively; 2,038 (1.30%) and 116 (0.07%) of 156,671 lots were positive for Salmonella and E. coli O157:H7, respectively; and 59 (0.36%) of 16,515 lots were positive for non-O157 Shiga toxin–producing E. coli. For ground beef, 46 (0.10%), 27 (0.06%), and 19 (0.04%) of 45,769 sublots exceeded CLs for SPC organisms, coliforms, and E. coli, respectively; and 329 (1.40%) and 18 (0.08%) of 23,475 sublots were positive for Salmonella and E. coli O157:H7, respectively. All lots and sublots found to exceed indicator organism CLs or to contain pathogens were identified, rejected for purchase, and diverted from federal nutrition assistance programs.

HIGHLIGHTS
  • AMS purchases beef for the National School Lunch Program.

  • Less than 0.10% of beef samples exceeded indicator organism critical limits.

  • Salmonella was found in 1.4% and E. coli O157:H7 was found in 0.08% of samples.

  • Indicator critical limit exceedance was weakly associated with pathogen presence.

  • Beef with excessive indicator organisms or containing pathogens was rejected for purchase.

Research -Antibacterial Interactions of Colloid Nanosilver with Eugenol and Food Ingredients

Journal of Food Protection

ABSTRACT

This study was conducted to investigate antibacterial properties of the colloidal silver nanoparticles (SNPs) and eugenol, alone and in combination, on Staphylococcus aureus and Salmonella Typhimurium and their interactions with food constituents (fat, protein, and carbohydrate). We examined antibacterial activities of SNPs and eugenol in Luria-Bertani (LB) broth and 1.5 and 3% fat ultrahigh-temperature (UHT) milk. MICs of eugenol and SNPs (particle size of 31.3 nm) were also investigated in the presence of sunflower oil, meat extract, and starch at concentrations of 2, 5, and 10% to examine the interactions between food constituents and antimicrobial agents. MICs and MBCs of eugenol and SNPs for both bacteria were at 2,500 and 25 μg/mL, respectively. Combinations of the two substances had additive and synergistic effects on Salmonella Typhimurium and S. aureus, respectively. Both compounds had bactericidal activity. In food matrices, results indicated that eugenol only in sunflower oil at 5 and 10% concentrations had significant antibacterial activity. A similar result was achieved for SNPs with 10% meat extract. In LB broth, eugenol at 2,500 and 5,000 μg/mL achieved 6-log reductions in the microbial population of both bacteria after 3 h, while SNPs achieved the same effect after 9 h. In UHT milk with 1.5% fat, eugenol at 5,000 μg/mL and SNPs at 25 μg/mL achieved 6-log reductions in bacterial populations after 24 h. Thus, the antimicrobial activity of both eugenol and SNPs depended on the medium in which the experiment was conducted, and the combination of both antimicrobial agents increased the antimicrobial effect.

HIGHLIGHTS
  • The interactions of eugenol, nanoparticles, and food constituents were investigated.

  • Eugenol and SNP had synergistic effects on S. aureus.

  • Protein and lipids reduced the efficacy of eugenol.

  • Protein negatively impacted the activity of SNPs.

Research – Antifungal Activity of Selected Natural Preservatives against Aspergillus westerdijkiae and Penicillium verrucosum and the Interactions of These Preservatives with Food Components

Journal of Food Protection

ABSTRACT

The present study examined the influence of primary food components on the antifungal activity of the essential oil of Origanum vulgare, carvacrol, thymol, eugenol, and trans-cinnamaldehyde against Penicillium verrucosum and Aspergillus westerdijkiae. The MIC was determined in food model media enriched with proteins (1, 5, or 10%), carbohydrates (1, 4, or 6%), or oil (1, 5, or 10%). Proteins increased the antifungal activity of O. vulgare essential oil, carvacrol, thymol, and eugenol, whereas the effect of trans-cinnamaldehyde decreased with increasing protein content. The presence of carbohydrates diminished the inhibitory effect of the natural preservatives on A. westerdijkiae; for P. verrucosum, their inhibitory effect increased with carbohydrates. Only the antifungal activity of trans-cinnamaldehyde did not depend on the carbohydrate content. The presence of oil had the strongest influence. At a concentration of 1% oil, the antifungal activity decreased significantly, and at 10% oil, almost no inhibition was observed. To investigate the effect of the antifungal agents on the morphology of the target molds, they were grown on malt extract agar containing carvacrol and trans-cinnamaldehyde and were examined by scanning electron microscopy. The hyphae, conidiophores, vesicles, and phialides were severely altered and deformed, and spore formation was clearly suppressed.

HIGHLIGHTS
  • The antifungal activity of natural preservatives is influenced by the food matrix.

  • Proteins increase the impact of O. vulgare EO, carvacrol, thymol, and eugenol.

  • Carbohydrates diminish the inhibition of natural preservatives on A. westerdijkiae.

  • In the presence of oil, natural inhibitors lose their antifungal effect.

  • Carvacrol and trans-cinnamaldehyde lead to distorted hyphae and loss of sporulation.

Research – Inactivation of Listeria monocytogenes, Staphylococcus aureus, and Salmonella enterica under High Hydrostatic Pressure: A Quantitative Analysis of Existing Literature Data

Journal of Food Protection

ABSTRACT

High hydrostatic pressure processing (HPP) is a mild preservation technique, and its use for processing foods has been widely documented in the literature. However, very few quantitative synthesis studies have been conducted to gather and analyze bacterial inactivation data to identify the mechanisms of HPP-induced bacterial inactivation. The purpose of this study was to conduct a quantitative analysis of three-decimal reduction times (t) from a large set of existing studies to determine the main influencing factors of HPP-induced inactivation of three foodborne pathogens (Listeria monocytogenes, Staphylococcus aureus, and Salmonella enterica) in various foods. Inactivation kinetics data sets from 1995 to 2017 were selected, and t values were first estimated by using the nonlinear Weibull model. Bayesian inference was then used within a metaregression analysis to build and test several models and submodels. The best model (lowest error and most parsimonious) was a hierarchical mixed-effects model including pressure intensity, temperature, study, pH, species, and strain as explicative variables and significant factors. Values for t and ZP associated with inactivation under HPP were estimated for each bacterial pathogen, with their associated variability. Interstudy variability explained most of the variability in t values. Strain variability was also important and exceeded interstudy variability for S. aureus, which prevented the development of an overall model for this pathogen. Meta-analysis is not often used in food microbiology but was a valuable quantitative tool for modeling inactivation of L. monocytogenes and Salmonella in response to HPP treatment. Results of this study could be useful for refining quantitative assessment of the effects of HPP on vegetative foodborne pathogens or for more precisely designing costly and labor-intensive experiments with foodborne pathogens.

HIGHLIGHTS
  • A meta-analysis was performed to identify factors influencing HPP inactivation of pathogens.

  • Three-decimal reduction times following HPP were estimated from existing data.

  • Staphylococcus aureus is the most piezoresistant of the three pathogens studied.

  • These three foodborne pathogens are less HPP resistant in acidic products.