Category Archives: Microbiology

Research – Efficacy of Acetic Acid or Chitosan for Reducing the Prevalence of Salmonella- and Escherichia coli O157:H7–Contaminated Leafy Green Plants in Field Systems

Journal of Food Protection

Outbreaks associated with fresh-cut leafy greens continue to occur despite efforts to implement horticultural practices that minimize introduction of enteric pathogens to the crop. The experimental trials in this study were designed to examine the efficacy of an acetic acid (AA)- and chitosan-based spray treatment, applied 1 day prior to harvest, for reducing the prevalence of Escherichia coli O157:H7 (O157) and Salmonella in field-grown leafy greens contaminated at levels detectable only through enrichment culture. Responses to the treatment solution were variable and depended on the type of leafy green (leafy lettuce, spinach, or cabbage), cultivar, pathogen, and AA concentration (0.3 to 0.7%). No significant differences in E. coli O157 prevalence were found for untreated and treated cabbage heads and spinach plants (P > 0.05). In contrast, treatment significantly affected Salmonella on ‘Bravo F1’ green cabbage and ‘7-Green’ spinach (P < 0.05), with odds ratios of 2.2 and 3.3 for finding the pathogen on untreated versus treated greens, respectively. Salmonella was also 7.1 times more likely to be found on an untreated lettuce plant than on a lettuce plant sprayed with a 0.7% AA treatment solution (95% confidence interval [CI], 4.1 to 12.2; P < 0.0001). In studies addressing the efficacy of chitosan (0.1 or 0.3%), this chemical failed to reduce the prevalence of either pathogen on lettuce (P > 0.05). Similarly, spraying with 0.3% AA did not affect the prevalence of Salmonella on lettuce plants (P > 0.05); however, treatment solutions with 0.4% AA reduced the likelihood of detecting Salmonella in treated versus untreated plants by 6.6 times (95% CI, 2.1 to 20.9; P = 0.0007). After the lettuce was harvested and hand washed, consumers failed to distinguish either visually or organoleptically between untreated lettuce and lettuce sprayed with an acetic acid solution (P > 0.05). These results indicate that acetic acid could be used to reduce the microbiological risk of preharvest leafy greens.

USA – FDA takes steps to improve ice cream production facility safety following inspections and environmental sampling efforts

FDA

“Inspecting food facilities and collecting and testing samples from the environment where foods are produced are two of the many ways the FDA works to better understand microbial hazards and to help prevent contaminated products from reaching consumers. These activities help the FDA gather data and information necessary to develop prevention-based systems and, when contamination does occur, to respond swiftly to these hazards. Following a string of safety issues related to a number of U.S. ice cream distributors, the FDA engaged a team to inspect and obtain environmental samples from 89 ice cream production facilities in 32 states to test for Listeria monocytogenes and Salmonella. Although many of these facilities were adhering to good manufacturing practices, we did find that some were in violation of the law,” said Frank Yiannas, FDA Deputy Commissioner for Food Policy and Response. “Findings from our inspections resulted in three voluntary recalls that were conducted in 2017 and 2018 to protect public health. We also collaborated with inspected companies to help them make needed corrections and implement food safety plans designed to keep harmful bacteria out of their products and protect American consumers. These results serve as an important reminder to all food facilities distributing products in the U.S. of the importance of complying with rules set forth to mitigate safety issues. Ultimately, we must work together to ensure all necessary protective steps are taken so that Americans can continue having confidence that the foods available for purchase in the U.S. are safe and wholesome.”

Research – Effects of Package Atmosphere and Storage Conditions on Minimizing Risk of Escherichia coli O157:H7 in Packaged Fresh Baby Spinach

Journal of Food Protection

ABSTRACT

Packaged fresh spinach has been associated with outbreaks of illness caused by Escherichia coli O157:H7. The purpose of this study was to assess the behavior of E. coli O157:H7 in packaged baby spinach in response to storage conditions of temperature and package atmosphere and including effects of inoculation level, spinach leaf damage (cut leaves), internalized or leaf surface contamination, exposure to hypochlorite sanitizer, and package size. Behavior of E. coli O157:H7 inoculated at 2 and 4 log CFU/g on spinach packaged in polymer bags composed of a two-layer laminate (polypropylene and polyethylene) and stored under atmospheres of 20% O2–3% CO2 and 0% O2–15% CO2 (aerobic and anaerobic, respectively) was assessed at 5, 7, 12, and 15°C for up to 14 days. Growth kinetics were calculated using DMFit software. Temperature decreases progressively diminished growth or survival of the pathogen, and an aerobic package atmosphere resulted in longer lag times (4 to 6 days) and lower population levels (0.2 to 1.4 log CFU/g) compared with the anaerobic atmosphere at 15°C. Internalized contamination, leaf cuts, or exposure to 100 ppm of hypochlorite did not result in changes in pathogen behavior compared with controls; however, a growth minimization trend consisting of longer lag times and lower population levels was repeatedly observed in the aerobic compared with the anaerobic package atmospheres. In contrast, growth of indigenous mesophiles and Enterobacteriaceae was unaffected by package atmosphere. Spinach stored at 5 to 7°C in two sizes (5 and 16 oz) of polyethylene terephthalate clamshell packages with ambient air atmospheres was more likely to progress to lower-oxygen conditions in 16-oz compared with 5-oz packages after 7 days of storage (P < 0.05). Practices to maintain aerobic conditions within the package, as well as storage of the package at low temperature, are ways to limit growth of E. coli O157:H7 in packaged spinach.

HIGHLIGHTS
  • Cold aerobic conditions limited survival of E. coli O157:H7 in packaged spinach.

  • Low-oxygen atmosphere increased pathogen risk in temperature-abused packages.

  • Internalization, leaf cuts, and hypochlorite stress did not increase pathogen risk.

  • Large spinach packages trended toward lower-oxygen conditions more than small packages.

  • Maintaining cold aerobic conditions can limit pathogen risk in packaged spinach.

Research – Antibacterial Effects of Phytic Acid against Foodborne Pathogens and Investigation of Its Mode of Action

Journal of Food Protection

This study investigated the antimicrobial mechanism of phytic acid (PA) and its antibacterial effects in combination with ethanol. The MIC of PA on Escherichia coli ATCC 11229, Staphylococcus aureus ATCC 6538P, Bacillus subtilis ATCC 6633, and Salmonella Typhimurium CICC 27483 were 0.24, 0.20, 0.26, and 0.28% (w/w), respectively. E. coli ATCC 11229 and S. aureus ATCC 6538P were selected to investigate the mechanism of PA by analyzing its effects at 1/2MIC and at MIC on the cell morphology, intracellular ATP, and cell membrane integrity. Environmental scanning electron microscope images revealed that PA was able to change the cell morphology and disrupt the intercellular adhesion. PA retarded bacterial growth and caused cell membrane dysfunction, which was accompanied by decreased intracellular ATP concentrations. Flow cytometry analysis further revealed that almost all the bacterial cells were damaged after treatment with PA at its MIC for 2 h. Moreover, PA has a synergistic antimicrobial ability when used in combination with ethanol. These results suggested that PA is effective in inhibiting growth of foodborne pathogens mainly by the mechanism of cell membrane damage and to provide a theoretical basis for the development of natural antimicrobial agents in the food industry.

Research – Aflatoxins in almonds: Monitoring and exposure assessment

Wiley Online

Abstract

Aflatoxins (AFs) are naturally occurring contaminants classified as human carcinogens. This article describes presence and concentrations of AFs in both raw and roasted almonds collected from Turkey from September 2017 to February 2018. Long‐term exposure to AFs was then calculated using the per capita consumption of almonds and the concentration of AFs in almonds. AFs were determined by high performance liquid chromatography coupled to fluorescence detector (HPLC‐FLD). The analytical method fulfilled the requirements on method performance according to Commission Regulation (EC) No 401/2006. AFs were detected in 15% of almonds, but at levels far below the European Union (EU) maximum levels (MLs). The long‐term mean exposure to AFB1 and total AFs via the consumption of almonds for Turkish consumer were calculated as 0.0019 and 0.0021 ng kg−1 b.w. day−1 at the lower bound, and 0.0043 and 0.0067 ng kg−1b.w. day−1 at the upper bound, respectively.

Practical applications

The present work was focused on aflatoxins (AFB1, AFB2, AFG1, and AFG2) contamination levels of raw and roasted almond kernels consumed in Turkey. The exposure assessment of general Turkish population to AFs through the consumption of almonds was also assessed. This study underlines the importance of periodic monitoring studies to provide a high level of food safety. This is the first report on exposure to AFs through the consumption of almond kernels for Turkish consumers.

Research – Comparison of Antimicrobial Treatments Applied via Conventional or Handheld Electrostatic Spray To Reduce Shiga Toxin–Producing Escherichia coli on Chilled Beef Outside Rounds

Journal of Food Protection

The purpose of this study was to compare the efficacy of different antimicrobial interventions applied via either conventional spray (CS) or handheld electrostatic spray (ESS) to reduce Shiga toxin–producing Escherichia coli (STEC) on fresh beef surfaces. Hot-boned outside rounds (ORs) were inoculated within 1 h after harvest with a cocktail of eight isolates consisting of 8 O157 and non-O157 serogroups of STEC (STEC8). ORs were hung on sterile meat hooks at 4°C for 36 h to simulate a contaminated full carcass side in the chiller. ORs were then treated with lactic acid (LA; 4.5%, w/v), 3.0% lauric arginate ester (LAE), 0.8% cetylpyridinium chloride, 200 mg/L peracetic acid, 3 mg/L chlorine dioxide, 5 mg/L ClO2, or tap water by using CS or ESS. Temperatures of LA and peracetic acid were set at 55 and 42°C before spraying, whereas all other solutions were applied at room temperature (25°C). Pretreatment and posttreatment STEC8-inoculated beef tissue samples were aseptically collected to evaluate the efficacy of interventions by application method (CS or ESS). LA applied with CS achieved the greatest reduction in STEC8 numbers (3.3 log CFU/cm2) compared with all other treatments: 0.2 log CFU/cm2 (tap water) to 2.3 log CFU/cm2 (LAE). Only for LA did a significant difference arise in mean STEC8 reductions between CS and ESS applications (3.2 versus 1.7 log CFU/cm2, respectively). Among the treatments applied with ESS, LAE produced the greatest reduction of STEC8. Antimicrobial interventions applied via conventional wand or cabinet-applied technologies can reduce the O157 and non-O157 STEC on fresh beef carcass surfaces, reducing transmission to beef consumers.

HIGHLIGHTS
  • We found no advantage in the use of electrostatic spray to reduce STEC8 on cold beef.

  • Greatest reductions in STEC8 were achieved by lactic acid with conventional spray.

  • Lauric arginate ester was the second best antimicrobial agent at reducing STEC8.

  • Lactic acid reduced pH on the beef surface significantly.

  • There was no effect of antimicrobial solution on temperature increase on beef outside rounds.

Research – Relationship of Sanitizers, Disinfectants, and Cleaning Agents with Antimicrobial Resistance

Journal of Food Protection

Sanitizers, disinfectants, and cleaning agents are vital to food hygiene assurance and are a major public health protection measure. Limiting microbial antibiotic resistance is also a global public health priority. Although many factors contribute to the rise in antimicrobial resistance in bacteria infecting humans, antibiotic use in both human clinical settings and for food-producing animals are primary contributors. Some concerns have been raised about the possibility of coselection between food hygiene chemicals and reduced antimicrobial susceptibility. This article reviews available evidence from individual studies purporting to demonstrate a possible risk of antimicrobial resistance development, following biocide usage. Furthermore, the conclusions of several key expert reports and meta-analysis publications were assessed for supportive evidence of a relationship between biocide usage in food production and resistance development. Although many studies report on the isolation of antimicrobial-resistant bacterial strains in food, evidence is lacking on the attribution of this resistance to biocide usage. Also, although a theoretical risk of causality exists, many of the studies performed to demonstrate this are in vitro studies using laboratory-grown or -trained bacterial isolates, challenged with sublethal (below the recommended food industry) disinfectant or sanitizing agent concentrations. The proper use of, and adherence to biocide manufacturer’s instruction for use, and the avoidance of biocide active agent dilution (e.g., through biofilm presence) must be ensured in food production environments. It is recommended that in situ studies should be performed to further assess causality, ensured a clear differentiation between interpretation of stable antimicrobial resistance and phenotypic adaptation. Furthermore, authorization of new biocidal active substances should take a scientific and risk-based approach regarding the potential for driving microbial resistance.

HIGHLIGHTS
  • Sanitizers and disinfectants (biocides) are essential for food safety assurance.

  • Concerns have been raised about theoretical risk of biocide-induced antimicrobial resistance.

  • In vitro studies provide weak causal evidence to attribute antimicrobial resistance to biocide usage.

  • GMPs, proper biocide usage, and avoidance of biofilms mitigate risk of antimicrobial resistance.

RASFF Alert – Animal Feed -Enterobacteriaceae – Pet Food

RASFF-Logo

RASFF – too high count of Enterobacteriaceae (up to 1310 CFU/g) in pet food from China in Sweden

 

Research – Limitation of microbial spoilage of rainbow trout fillets using characterized thyme oil antibacterial nanoemulsions

Wiley Online

Abstract

Thyme oil nanoemulsions (TONa and TONb), having 219 nm and 163 nm diameters, were successfully prepared by using sonication technique. Zeta potential (ZP) and polydispersity indexes (PDI) of TONa and TONb were defined to be 19.77, 0.24 and −24.80, 0.054, respectively. It was determined that zeta size values of TONs might have played a role on the inhibition of Pseudomonas aeruginosa, Escherichia coli, Salmonella typhimurium. All bacteria tested by agar diffusion method were gram‐negative bacteria that could be mostly found in fish fillets. Within 9 days cold storage period, TMAB growth in fish fillets treated with TONb was successfully reduced from 6.42 to 4.62 log CFU/g (change: 28%). In addition, as compared with control group samples, TPB growth of the same group was limited by 1.51 log CFU/g (from 7.93 to 6.42 log CFU/g). The results suggested that treatment of the fish fillets with the TONs would be a useful and practical nano application to delay microbial growth in fish fillets.

Practical applications

Thyme oil nanoemulsions (TONa and TONb) with a lower diameter than 219 nm have been successfully obtained. Antibacterial effects of TONa and TONb nanoemulsions have been revealed before treatment with fish fillets. The results revealed that TONs could be successfully used for the limitation of microbial spoilage of fish fillets stored at 4°C for 9 days. In this respect, the nano application with rapid and cost‐effective has been presented for the fishery processing industry.

Research – Exploratory Study of the Application of Smoke Aerosols to Manure-Based Composting Materials To Reduce Prevalence of Salmonella

Journal of Food Protection

During the early stages of aerobic composting, heat is generated and when the materials are self-insulating, extended exposure of pathogens to this heat source will lead to significant reduction, if not elimination, of the pathogens. However, when insufficient heat is applied to the composting materials, pathogens may survive. Under those conditions if the compost had contained material of animal origin or food waste, it would be considered untreated and would not be allowed in fields growing crops that may be consumed raw. However, alternative treatment processes are allowed, provided they are validated to meet the microbial standards stipulated in the Produce Safety final rule of the Food Safety Modernization Act and that the physical parameters of the process are documented to ensure that the conditions under which the process was validated have been met. Hence, this exploratory study was undertaken in a laboratory setting to determine the potential for application of aerosolized smoke to inactivate Salmonella in manure-based compost. Smoke generated from wood chips (oak or pecan) and introduced to the headspace of contaminated cow manure compost (≤3 log CFU/g) in sealed containers (35 g per container) resulted in no Salmonella detected by enrichment culture in 100% (0 of 14) of the samples after 18 to 48 h of exposure, whereas Salmonella in control samples remained at initial levels over the same time period. Shorter exposure times (6 h) to the smoke aerosols were less effective (11 of 24 samples positive by enrichment culture), and additional flushes with the wood smoke during this time failed to decrease the prevalence of contamination. Smoke aerosols generated from waste agricultural materials and held in containers with Salmonella-contaminated compost for 18 h significantly reduced the prevalence of the pathogen in samples compared with control samples (P < 0.05). The odds of not finding Salmonella in smoke-exposed compost were 14 (pine needles and rice hulls), 23 (cocoa hulls, orange rind, and peanut hulls), and 28 (sunflower hulls) times greater compared with samples not exposed to smoke. Many other variables remain to be examined (e.g., compost composition, compost maturity, and anaerobic conditions) to determine whether this approach could be universally applied to manure-based compost. Validation under field conditions will be required and may entail use of this approach in combination with suboptimal thermal conditions (<55°C).