Category Archives: Research

Research -Evaluation of the Efficacy of Three Direct Fed Microbial Cocktails To Reduce Fecal Shedding of Escherichia coli O157:H7 in Naturally Colonized Cattle and Fecal Shedding and Peripheral Lymph Node Carriage of Salmonella in Experimentally Infected Cattle

Journal of Food Protection

ABSTRACT

Two experiments were conducted to evaluate the feeding of direct fed microbials (DFMs) on fecal shedding of Escherichia coli O157:H7 in naturally infected cattle (experiment I) and on Salmonella in the feces and peripheral lymph nodes (PLNs) of experimentally infected cattle (experiment II). Thirty cattle, 10 per treatment, were used in each experiment. Treatments in experiment I consisted of a control (lactose carrier only); DFM1, a 1:1 ratio of Enterococcus faecium and Lactobacillus animalis; and DFM2, a 1:1 ratio of Lactobacillus acidophilus and Pediococcus acidilactici. In Experiment II, DFM1 was replaced with DFM3, a 1:2 ratio of Lactobacillus reuteri and other Lactobacillus strains. Additives were mixed in water and applied as a top-dressing to each pen’s daily ration for 50 days. Approximately half-way through each experiment, the DFM concentration was doubled for the remainder of the study. Fecal samples were collected throughout experiment I and cultured for E. coli O157:H7. Cattle in experiment II were inoculated intradermally with Salmonella Montevideo on days 32, 37, and 42 and then necropsied on days 49 and 50 (five cattle per treatment on each day). Innate immune function was assessed on days 29, 49, and 50. In experiment I, fecal concentration and prevalence of E. coli O157:H7 were not different (P > 0.10) nor was there an effect (P = 0.95) on the percentage of super shedders (cattle shedding ≥3.0 log CFU/g of feces). In experiment II, no treatment differences (P > 0.05) were observed for Salmonella in the PLNs except for the inguinal nodes, which had a significantly lower Salmonella prevalence in DFM-supplemented cattle than in the controls. Immune function, as measured by monocyte nitric oxide production and neutrophil oxidative burst, was decreased (P < 0.05) in the DFM treatment groups. Although results of this research indicate little to no effect of these DFMs on E. coli O157:H7 or Salmonella in cattle, an increase in the duration of administration to that similar to what is used for commercial cattle might elicit treatment differences.

HIGHLIGHTS
  • Direct fed microbials were evaluated for pathogen mitigation in cattle.

  • No treatment effects on fecal shedding of E. coli O157:H7 were observed.

  • DFM treatment reduced Salmonella in only one the four lymph node types examined.

  • Under these experimental conditions, short-term feeding of DFMs failed to mitigate pathogens.

 

Research – Effect of Eryngium caeruleum essential oil on microbial and sensory quality of minced fish and fate of Listeria monocytogenes during the storage at 4°C

Wiley Online

The present study investigated in vitro antimicrobial activity of Eryngium caeruleum essential oil (EEO) against five foodborne pathogenic bacteria based on microdilution and disk diffusion methods. Moreover, its effects on specific spoilage microorganisms, inoculated Listeria monocytogenes, and its sensory changes in minced fish were evaluated during 12 days of storage at refrigeration temperature. The results showed that Staphylococcus aureus and Escherichia coli were the most sensitive and the most resistant bacteria with a minimum inhibitory concentration of 0.125 and 1 mg/ml, as well as inhibition zones of 15.66 and 11.66 mm, respectively. Regarding the antimicrobial effect of EEO on the microbial profile and inoculated L. monocytogenes, treating with 0.4% EEO caused a significant decrease in the studied microorganisms when compared to the control group (p < 0.05). In addition, considering the sensory evaluation, the best scores were observed for the samples treated with 0.2% and 0.4% EEO. However, none of the groups obtained acceptable scores until the final day of storage except for the color attribute. In general, sensory evaluation and its correlation with microbial counting indicated that the treatment with 0.4% EEO was able to preserve the microbial quality of the minced fish at refrigeration temperature without any undesirable sensory effects.

Research – Thermal and Chemical Treatments To Reduce Salmonella on Alfalfa (Medicago sativa) and Broccoli (Brassica oleracea var. italica) Seeds before and during the Sprouting Process: A Hurdle Approach

Journal of Food Protection

ABSTRACT

Sprouts are vehicles of foodborne diseases caused by pathogens such as Salmonella. The aim of this study was to evaluate thermal and chemical treatments applied as a hurdle approach to reduce Salmonella in alfalfa (Medicago sativa) and broccoli (Brassica oleracea var. italica) seeds before and during their germination. Seeds, inoculated and then dried at 55°C for 48 h, were subjected to a chemical treatment and a thermal shock with (i) 75 mM caprylic acid at 70°C for 5 s, (ii) 0.04% CaO at 70°C for 5 s, or (iii) 1% H2O2 at 70°C for 5 s. After each treatment, seeds were immersed in water at 3°C for 5 s. Next, the imbibition process was carried out with 0.016% H2O2 at pH 3.0. Finally, the seeds were transferred to a rotary drum-type germinator and were sprayed with the same chemical solution that was applied before the imbibition process, for 20 s at intervals of 5 min for 40 min at 3 rpm. All chemical treatments reduced Salmonella at least 5 log CFU/g on both seeds. Germination rates between 90 and 93% were obtained after application of thermal and chemical treatments. Salmonella was not detected after the imbibition stage when caprylic acid and H2O2 treatments were applied. However, during the germination process of both seeds, Salmonella counts of >6 log CFU/g were obtained despite all treatments being applied at different stages of the sprouting process. These results demonstrated that thermal and chemical treatments used as a hurdle approach to control Salmonella on alfalfa and broccoli seeds significantly reduced the pathogen concentration on seeds >5 log but were ineffective to eliminate Salmonella and to control its growth during the sprouting process. The production of safe sprouts continues to be a major challenge for industry.

HIGHLIGHTS
  • A hurdle approach reduces Salmonella concentration >5 log on alfalfa and broccoli seeds.

  • Germination rates over 90% were reached on alfalfa and broccoli seeds after sequential treatments.

  • Salmonella counts increase during germination, despite the efficacy of previous treatments on seeds.

  • Production of safe sprouts continues to be a major challenge for industry.

Research – Foodborne Illness and the Struggle for Food Safety

CDC

Professor Timothy D. Lytton, a keen scholar of regulatory evolution, provides a lively and well-documented guide to 150 years of major advances in food safety regulation and prevention in the United States. He starts with the early efforts to cleanse and regulate the milk supply in the 19th century that ultimately led to near-universal pasteurization. Efforts to make canned food free of botulism in the 1920s led to a new focus on critical control steps in processing, using sufficient time and heat to eliminate the risk, and thus to a new general approach based on process control. Modernizing meat inspection with process control logic in the 1990s and the recent efforts to make fresh produce safer in the 2000s take the reader to the controversies of the present day.

This book fills a critical gap, weaving the history of public health, regulatory agencies, and the food industry together with issues of immediate concern today. It is an innovative perspective that captures the complexity of the system beyond the scientific report or published regulation. The book should be of interest to students and practitioners of public health and food science and anyone interested in making food reliably safe.

With fresh examples and detailed interviews, Lytton illustrates the dynamic interplay of outbreak investigations, better prevention strategies developed by industry, consumer advocacy, and regulations. He explains why the resulting balance is a punctuated equilibrium, with longer steady states ending in momentous rapid change. Large and catastrophic outbreaks come as the final trigger, as “focusing events” that, with media coverage, increase public attention and create pressure for change. Lytton tells the striking and less well-known story of what happens behind the scenes as food safety champions within the industry push new solutions and voluntary standards forward, show how they could reduce contamination, and gain adherents up and down the food supply chain, thus leading the way for others in industry and regulators to follow. He also deftly outlines the complex roles of third-party auditors, who provide information to one company about the safety practices of its suppliers, and provides a fresh perspective on the growing role that liability insurers may play in the future.

This is history that uplifts, showing how we honor those who suffered from and died of a foodborne disease that is now preventable in the form of better practices and safer food today. In the crucible of public action, it reminds us all how these advances begin and, with feedback and learning, how they can succeed.

Research – Prevalence of Salmonella in cucumbers, antibiotic and acid resistances and description of the kinetic behavior with dynamic model during storage

Wiley Online

This study isolated Salmonella from cucumbers, analyzed the antibiotic resistance and acid resistance for the isolates and developed a dynamic model. Salmonella prevalence in cucumbers and their resistances were determined. To describe the kinetic behavior of Salmonella isolates, the isolates were inoculated into cucumbers, and Salmonella cell counts were enumerated during storage at 10–30°C. The Baranyi model was fitted to the cell count data to calculate kinetic parameters (lag phase duration [LPD] and maximum specific growth rate (μmax)], and a polynomial model was fitted to the kinetic parameters as a function of temperature. The model performance was evaluated with root mean square error (RMSE). Using these models, a dynamic model was developed. Salmonella were detected in 3 of 24 cucumbers, all of which were multidrug‐resistant and one was acid‐resistant. As storage temperature increased, LPD decreased and μmax increased. These models were appropriate with 0.367 of RMSE. These results suggest that cross‐contaminated Salmonella could increase during transportation, and it may lead to human infection.

Research – Acidic environment could boost power of harmful pathogens

Science Daily

When food we’ve swallowed reaches our stomachs, it finds an acidic environment. The low pH in the stomach helps to begin digestion — and has been thought to kill the bacteria that hides in food that otherwise could harm our bodies.

However, recent work from the Ackley and Chandler labs in the Department of Molecular Biosciences at the University of Kansas runs counter to this idea, instead suggesting lower pH in the digestive tract may make some bacterial pathogens even more harmful.

Their findings, published in the peer-reviewed journal PLOS Pathogens, could have implications for addressing the crisis of antibiotic resistance in bacterial infections around the world.

The investigation was performed using small, bacteria-eating organisms called Caenorhabditis elegans.

Research – Treatment of fresh produce with a Salmonella‐targeted bacteriophage cocktail is compatible with chlorine or peracetic acid and more consistently preserves the microbial community on produce

Wiley Online

Diets rich in minimally processed foods are associated with numerous health benefits, in part, due to their diverse, natural microbiota. However, antimicrobials, such as chlorine and peracetic acid (PAA), that are used to address food safety concerns may damage the natural microflora of fresh produce. One promising approach for targeting pathogenic bacteria in foods without impacting the normal food microbiota are bacteriophages. In this study, we observed that combinational treatment of conventional antimicrobials (PAA and chlorine) and bacteriophages, specifically the Salmonella‐targeted preparation SalmoFresh, retained the bactericidal effectiveness of individual interventions, and in some cases, achieved substantially increased efficacy. Additionally, the bacterial microbiomes of farm fresh and organic produce were less affected after phage treatment compared to PAA and chlorine. Finally, our study revealed that resistance rates against SalmoFresh were relatively minor and unaffected by the stresses introduced after chemical washes and/or bacteriophage treatment.

Research-Nisin-Based Organic Acid Inactivation of Salmonella on Grape Tomatoes: Efficacy of Treatment with Bioluminescence ATP Assay

Journal of Food Protection

ABSTRACT

The antimicrobial activity of a new nisin-based organic acid sanitizer (NOAS), developed in our laboratory, was tested against viable aerobic mesophilic bacteria and Salmonella populations inoculated on produce surfaces. The activity of NOAS was compared with 200 ppm of chlorinated wash water and a bioluminescence ATP technique to determine the efficacy of treatments compared with plate count methods. The activity of the 10% final concentration of NOAS against viable populations of 109 CFU/mL Salmonella in phosphate-buffered saline (PBS), sterile deionized distilled water, and buffered peptone water was tested in vitro and on grape tomatoes inoculated with Salmonella at 2.5 log CFU/g. A similar batch of inoculated tomatoes were treated with 200 ppm of total available chlorinated water. All treatments for inactivation of viable Salmonella in vitro was tested up to 30 min and 5 min for the attached populations on tomatoes. Inactivation of viable Salmonella at 109 log CFU/mL by 10% the NOAS solution averaged >107 log CFU/mL in PBS, sterile deionized distilled water, and buffered peptone water. Similarly, Salmonella bacteria inactivated on tomato surfaces by the NOAS solution was significantly (P < 0.05) greater than numbers on chlorinated washed tomatoes, and surviving bacterial populations on NOAS and chlorine-treated tomatoes were <1 and 4 CFU/g, respectively. A significant linear correlation coefficient (r2 = 0.99) between the bioluminescence ATP assay and aerobic plate counts of inoculated and untreated grape tomatoes were recorded but not with NOAS and chlorine-treated tomatoes, as bacterial populations were less than the minimum baseline for determination. Also, the results indicated that the NOAS solution is a better alternative antimicrobial wash solution than 200 ppm of chlorinated water.

HIGHLIGHTS
  • The antimicrobial activity of NOAS was compared with chlorinated water.

  • Salmonella bacteria were more susceptible to NOAS than chlorinated water.

  • Assay correlated with aerobic plate count of inoculated and untreated tomatoes, not treated ones.

  • NOAS is an excellent alternative antimicrobial wash solution compared with chlorinated wash water.

Research – Inactivation of Murine Norovirus on Fruit and Vegetable Surfaces by Vapor Phase Hydrogen Peroxide

Journal of Food Protection

ABSTRACT

Vapor phase hydrogen peroxide (H2O2) can be utilized to inactivate murine norovirus (MNV), a surrogate of human norovirus, on surface areas. However, vapor phase H2O2 inactivation of virus on fruits and vegetables has not been characterized. In this study, MNV was used to determine whether vaporized H2O2 inactivates virus on surfaces of various fruits and vegetables (apples, blueberries, cucumbers, and strawberries). The effect of vapor phase H2O2 decontamination was investigated with two application systems. Plaque assays were performed after virus recovery from untreated and treated fresh produce to compare the quantity of infective MNV. The Mann-Whitney U test was applied to the test results to evaluate the virus titer reductions of treated food samples, with significance set at P ≤ 0.05. The infective MNV populations were significantly reduced on smooth surfaces by 4.3 log PFU (apples, P < 0.00001) and 4 log PFU or below the detection limit (blueberries, P = 0.0074) by treatment with vapor phase H2O2 (60 min, maximum of 214 ppm of H2O2). Similar treatments of artificially contaminated cucumbers resulted in a virus titer reduction of 1.9 log PFU. Treatment of inoculated strawberries resulted in 0.1- and 2.8-log reductions of MNV. However, MNV reduction rates on cucumbers (P = 0.3809) and strawberries (P = 0,7414) were not significant. Triangle tests and color measurements of untreated and treated apples, cucumbers, blueberries, and strawberries revealed no differences in color and consistency after H2O2 treatment. No increase of the H2O2 concentration in treated fruits and vegetables compared with untreated produce was observed. This study reveals for the first time the conditions under which vapor phase H2O2 inactivates MNV on selected fresh fruit and vegetable surfaces.

HIGHLIGHTS
  • Produce was treated with vapor phase H2O2 for 60 min (maximum of 260 ppm of H2O2).

  • A 4-log reduction in MNV was achieved by H2O2 treatment on apples and blueberries.

  • Reductions of MNV on treated strawberries and cucumbers were not significant.

Research- Survival Evaluation of Salmonella and Listeria monocytogenes on Selective and Nonselective Media in Ground Chicken Meat Subjected to High Hydrostatic Pressure and Carvacrol

Journal of Food Protection

ABSTRACT

High pressure processing (HPP) and treatment with the essential oil extract carvacrol had synergistic inactivation effects on Salmonella and Listeria monocytogenes in fresh ground chicken meat. Seven days after HPP treatment at 350 MPa for 10 min, Salmonella treated with 0.75% carvacrol was reduced to below the detection limit (1 log CFU/g) at 4°C and was reduced by ca. 6 log CFU at 10°C. L. monocytogenes was more sensitive to these imposed stressors, remaining below the detection limit during storage at both 4 and 10°C after HPP treatment at 350 MPa for 10 min following treatment with 0.45% carvacrol. However, pressure-injured bacterial cells may recover and lead to an overestimation of process lethality when a selective medium is used without proper justification. For HPP-stressed Salmonella, a 1- to 2-log difference was found between viable counts on xylose lysine Tergitol 4 agar and aerobic plate counts, but no significant difference was found for HPP-stressed L. monocytogenes between polymyxin–acriflavine–lithium chloride–ceftazidime–esculin–mannitol (PALCAM) agar and aerobic plate counts. HPP-induced bacterial injury and its recovery have been investigated by comparing selective and nonselective agar plate counts; however, few investigations have addressed this issue in the presence of essential oil extracts, taking into account the effect of high pressure and natural antimicrobial compounds (e.g., carvacrol) on bacterial survival in various growth media. Use of selective media may overestimate the efficacy of bacterial inactivation in food processing evaluation and validation studies, and the effects of various media should be systematically investigated.

HIGHLIGHTS
  • HPP and carvacrol had synergistic pathogen inactivation effects in ground chicken meat.

  • HPP at 350 MPa for 10 min with 0.60% carvacrol treatment resulted in a >5-log pathogen reduction.

  • A 1- to 2-log difference was found for counts of HPP-treated Salmonella on two growth media.

  • Counts of HPP-treated L. monocytogenes were similar on selective and nonselective media.

  • Carvacrol suppressed the growth and recovery of the HPP-treated bacterial cells.