Category Archives: Enterococcus

Research – Microbiological Quality and Safety of Fresh Turkey Meat at Retail Level, Including the Presence of ESBL-Producing Enterobacteriaceae and Methicillin-Resistant S. aureus



The aim of this work was to study the microbiological safety and quality of marketed fresh turkey meat, with special emphasis on methicillin-resistant S. aureus, ESBL-producing E. coli, and K. pneumoniae. A total of 51 fresh turkey meat samples were collected at retail level in Spain. Mesophile, Pseudomonas spp., enterococci, Enterobacteriaceae, and staphylococci counts were 5.10 ± 1.36, 3.17 ± 0.87, 2.03 ± 0.58, 3.18 ± 1.00, and 2.52 ± 0.96 log CFU/g, respectively. Neither Campylobacter spp. nor Clostridium perfringens was detected in any sample. ESBL-producing K. pneumoniae and E. coli were detected in 22 (43.14%), and three (5.88%) samples, respectively, all of which were multi-resistant. Resistance to antimicrobials of category A (monobactams, and glycilcyclines) and category B (cephalosporins of third or fourth generation, polymixins, and quinolones), according to the European Medicine Agency classification, was found among the Enterobacteriaceae isolates. S. aureus and methicillin-resistant S. aureus were detected in nine (17.65%) and four samples (7.84%), respectively. Resistance to antimicrobials of category A (mupirocin, linezolid, rifampicin, and vancomycin) and category B (cephalosporins of third- or fourth generation) was found among S. aureus, coagulase-negative staphylococci, and M. caseolyticus isolates.

Research – Microbiological safety of aged meat



The impact of dry-ageing of beef and wet-ageing of beef, pork and lamb on microbiological hazards and spoilage bacteria was examined and current practices are described. As ‘standard fresh’ and wet-aged meat use similar processes these were differentiated based on duration. In addition to a description of the different stages, data were collated on key parameters (time, temperature, pH and aw) using a literature survey and questionnaires.

The microbiological hazards that may be present in all aged meats included Shiga toxin-producing Escherichia coli(STEC),Salmonella spp., Staphylococcus aureus, Listeria monocytogenes, enterotoxigenic Yersinia spp., Campylobacter spp. and Clostridium spp. Moulds, such as Aspergillus spp. and Penicillium spp., may produce mycotoxins when conditions are favourable but may be prevented by ensuring a meat surface temperature of−0.5 to 3.0°C, with a relative humidity (RH) of 75–85% and an airflow of 0.2–0.5 m/s for up to 35 days.

The main meat spoilage bacteria include Pseudomonas spp., Lactobacillus spp. Enterococcus spp., Weissella spp., Brochothrix spp., Leuconostoc spp. Lactobacillus spp., Shewanella spp. and Clostridium spp. Undercurrent practices, the ageing of meat may have an impact on the load of microbiological hazards and spoilage bacteria as compared to standard fresh meat preparation. Ageing under defined and controlled conditions can achieve the same or lower loads of microbiological hazards and spoilage bacteria than the variable log10increases predicted during standard fresh meat preparation. An approach was used to establish the conditions of time and temperature that would achieve similar or lower levels of L. monocytogenes and Yersinia enterocolitica (pork only) and lactic acid bacteria(representing spoilage bacteria) as compared to standard fresh meat. Finally, additional control activities were identified that would further assure the microbial safety of dry-aged beef, based on recommended best practice and the outputs of the equivalence assessment.

Resaerch- Evaluation of Various Lactic Acid Bacteria and Generic E. coli as Potential Non-pathogenic Surrogates for In-Plant Validation of Biltong Dried Beef Processing


Validation studies conducted within a food processing facility using surrogate organisms could better represent the manufacturing process than controlled laboratory studies with pathogenic bacteria on precision equipment in a BSL-2 lab. The objectives of this project were to examine potential surrogate bacteria during biltong processing, conduct biltong surrogate validation lethality studies, and measure critical factors and intrinsic parameters during processing. Beef pieces (1.9 cm × 5.1 cm × 7.6 cm) were inoculated with four-strain mixtures of Carnobacterium divergens/C. gallinarumPediococcus acidilactici/P. pentosaceous, and Biotype 1 E. coli ATCC BAA (-1427, -1428, -1429, and -1430), as well as a two-strain mixture of Latilactobacillus sakei and other commercially available individual bacterial cultures (P. acidilactici Saga200/Kerry Foods; Enterococcus faecium 201224-016/Vivolac Cultures). Inoculated beef was vacuum-tumbled in marinade and dried in a humidity-controlled oven for 8–10 days (24.9 °C; 55% relative humidity). Microbial enumeration of surviving surrogate bacteria and evaluation of intrinsic factors (water activity, pH, and salt concentration) were performed post inoculation, post marination, and after 2, 4, 6, 8, and 10 days of drying. Trials were performed in duplicate replication with triplicate samples per sampling time and analyzed by one-way RM-ANOVA. Trials conducted with E. faeciumPediococcus spp., and L. sakei never demonstrated more than 2 log reduction during the biltong process. However, Carnobacterium achieved a >5 log (5.85 log) reduction over a drying period of 8 days and aligned with the reductions observed in previous trials with pathogenic bacteria (Salmonella, E. coli O157:H7, L. monocytogenes, and S. aureus) in biltong validation studies. Studies comparing resuspended freeze-dried or frozen cells vs. freshly grown cells for beef inoculation showed no significant differences during biltong processing. Carnobacterium spp. would be an effective nonpathogenic in-plant surrogate to monitor microbial safety that mimics the response of pathogenic bacteria to validate biltong processing within a manufacturer’s own facility. View Full-Text

Research – Determination of Listeria monocytogenes, competitive microflora, microbial hygiene indicators and physico-chemical parameters during the shelf-life of the typical Mediterranean style fermented sausage “Salsiccia Sarda”

Journal of Food Protection

The aim of the present study was the determination of Listeria monocytogenes , competitive microbiota, microbial hygiene indicators and physico-chemical parameters in the typical Mediterranean Style fermented sausages “Salsiccia Sarda” . A batch of “Salsiccia Sarda” (25 samples) naturally contaminated by L. monocytogenes and vacuum packaged after 24 days of ripening was included in the study. Fifteen samples stored at 8° C were analysed after 13, 90 and lastly at the end of shelf-life, after 180 days from vacuum packaging. Ten vacuum packaged samples were stored at 12°C in a domestic fridge simulating temperature abuse and were evaluated at the end of the shelf-life. Samples were subjected to physico-chemical analysis (pH and aw) and investigated for the presence and enumeration of L. monocytogenes . Competitive microbiota, Lactic Acid Bacteria (LAB) and Coagulase negative Staphylococci (CNS), and microbial hygiene indicators (Total mesophilic bacterial counts, Enterobacteriaceae, Enterococcuss spp. and Staphylococcus aureus ) were determined in all the samples. Although a decreasing trend in L. monocytogenes prevalence was observed through the shelf-life, the detection of the pathogen in fermented sausages confirms the ability of L. monocytogenes to overcome the hurdles of the manufacturing process. The results of the present study highlight the importance to carefully evaluate the “Salsiccia Sarda” production process by Food Business Operators (FBOs) in order to maintain unfavourable conditions for the growth of L. monocytogenes .

Research – New NARMS report shows rising resistance in Salmonella, Campylobacter


The latest data from a national surveillance system that monitors foodborne bacterial pathogens for antimicrobial resistance (AMR) shows some concerning changes in resistance patterns among serotypes of Salmonella.

The findings come from the National Antimicrobial Resistance Monitoring Systems (NARMS) 2019 Integrated Summary, which combines data from the Centers for Disease Control and Prevention (CDC), the Food and Drug Administration (FDA), and the US Department of Agriculture (USDA). The report provides a snapshot of resistance patterns found in bacteria isolated from humans, animals, raw meats from retail outlets (chicken, ground turkey, ground beef, and pork chops), and meat and poultry product samples collected at slaughtering facilities.

In addition to Salmonella, which causes an estimated 1.35 million illnesses and 26,500 hospitalizations each year, the NARMS report also includes resistance data on Campylobacter (1.5 million illnesses and 19,500 hospitalizations), Escherichia coli, and Enterococcus. NARMS monitors these bacteria to detect emerging resistance patterns to the antibiotics that are most important to human medicine, multidrug resistance, and specific resistance genes.

Increase in multidrug-resistant Salmonella serotype

Overall, the NARMS report shows that more than three fourths of the Salmonella isolates (78%) from humans were not resistant to any of the antibiotics tested, and that the overall level of resistance in humans remains relatively unchanged since 2018. However, the report also found rising resistance to ciprofloxacin—one of the three antibiotics used to treat severe Salmonella infections.

From 2018 to 2019, Salmonella with decreased susceptibility to ciprofloxacin increased from 9% to 11% in humans, from 18% to 31% in retail chicken, from 20% to 30% in chicken product samples, from 26% to 32% in chicken cecal content samples, and from 0% to 14% in retail pork samples.

The increase in resistance to ciprofloxacin among poultry isolates was primarily due to the increase in Salmonella Infantis, a multidrug-resistant (MDR) serotype that emerged in 2014.

The rise in Salmonella Infantis isolates was also behind an increase in MDR isolates found in retail chicken (from 20% to 32%) and in chicken product samples (22% to 29%). Up to 10 antimicrobial resistance (AMR) genes were found in some of the Salmonella Infantis isolates.

In addition, the NARMS report shows an increase in another MDR Salmonella serotype, I 4,[5],12:i:-, which is linked to pigs and has become an increasing public health concern in Europe and the United States. The percentage of MDR isolates from humans that are of the I 4,[5],12:i:- serotype rose from 7% in 2010 to 26% in 2019, and from 7% to 35% in swine samples.

NARMS data also show rising fluoroquinolone resistance in Campylobacter isolates. In humans, the proportion of ciprofloxacin-resistant Campylobacter isolates rose from 29% in 2018 to 34% in 2019 for Campylobacter jejuni and from 41% to 45% in C coli. Ciprofloxacin-resistant C jejuni isolated from chicken cecal contents (21% in 2018 to 26% in 2019) and chicken retail samples (20% to 22%) also rose.

Analysis of E coli isolates found increases in ceftriaxone resistance in sow cecal samples (3% in 2018 to 7% in 2019) and in retail pork (4% to 7%). Whole-genome sequencing of Salmonella and E coli from animals, animal products, and retail meats found that none harbored any of the MCR-1 through MCR-8 colistin-resistance genes.

Slovakia – Report on zoonoses, foodborne diseases and waterborne diseases in the Slovak Republic in 2020



The protection of human and animal health can only be achieved through the active cooperation of experts in the field of control and research in the human and veterinary field. The report on zoonoses, foodborne diseases and waterborne diseases in the Slovak Republic for 2020 contains data from official inspections carried out in the field of agriculture and health care, as well as from research institutes and universities. The preparation of the report was coordinated by the National Contact Point for Scientific and Technical Cooperation with the European Food Safety Authority (EFSA EFSA), which is established at the Department of Food Safety and Nutrition of the Ministry of Agriculture and Rural Development of the Slovak Republic (MPRV SR).

The report serves as a basis for the EFSA NCB and scientific experts to set priorities and own national food safety risk assessments. At the same time, the report serves as one of the bases for the Community risk assessment carried out by the European Food Safety Authority (EFSA). Scientific risk assessment is the basis for risk management. The report describes the situation in  35 zoonotic agents, 5 foodborne diseases (ie foodborne diseases) without zoonotic potential and 4 waterborne pathogens. Of the 44 agents monitored, 23 are bacterial, 10 parasitic, 10 viral and prion.

It presents the summary results of examinations and tests performed in 2020 in the Slovak Republic and the evaluation of the national epidemiological situation in humans and animals with a focus on trends and sources of zoonotic and foodborne diseases.

The report presents the summary results of examinations and tests carried out in 2020 and an assessment of the national epidemiological situation in humans and animals , focusing on trends and sources of zoonotic and foodborne diseases . The number of monitored authors, cooperating organizations and experts is growing every year. A wide team of more than 70 experts from 24 scientific and control organizations in the Slovak Republic took part in its elaboration .

In 2020, 17,067 human diseases caused by the study agents were reported, with 29.1% related to campylobacteriosis, 20.9% to Clostridium dificille and 20.4% to salmonellosis. Rotavirus 11.6%, Norwalk virus 5.1%, Borrelia burgdorferi sl 5.6% and  Escherichia coli 1.2% also contributed to a higher percentage of diseases.

Seven of the study agents caused 380 human epidemics, of which 56.6% were salmonellosis, 23.2% were campylobacteriosis and 12.6% of epidemics were caused by rotavirus. Norwalk virus accounted for 5.5%, tick-borne encephalitis virus 1.3%, shigella and 0.5% and yersinia 0.3%.  

35,957 food samples were examined for the presence of 15 pathogens with a positive finding in 2.2% of samples. Higher percentages of positive findings were in  Yersinia spp. 48.1%, Enterococcus spp. 46.3% and  Vibrio spp. 31.8%.

The presence of 30 pathogens was monitored in 2,483,239 samples originating from livestock and wild animals, pets and zoos taken as part of official control, preventive monitoring, research, as well as from sick or dead animals. Positive findings accounted for 0.1% of samples. Higher percentages of positive findings were recorded for  Aeromonas spp. 59.3%, Clostridium spp. 55.4%, Francisella tularensis 50.4%, Babesia spp. 41.1%, Dirofilaria spp. 34.8%, Campylobacter spp. 18.3%, Yersinia spp. 18.3%, Staphylococcus aureus 18.1%, hepatitis E virus 14.1%, Listeria monocytogenes  11.8%, Toxocara spp. 10.5%.

Feed – 385 samples were examined for the presence of Salmonella spp. (1.5% positive samples), Escherichia coli (60.0% positive samples) and Clostridium spp.

(1.7% positive samples).

35,746 water samples were examined for the presence of 9 agents, of which 7.4% were positive, of which Legionella spp. 47.1% and Vibrio spp. 39.5%.

44,633 samples from the environment were examined for the presence of 8 pathogens, of which 2.3% were positive, of which Legionella spp. 36.0%, Vibrio spp. 6.8%,  E.coli 3.6% and  Enterococcus spp. 2.6%.

The report also includes the results of examinations for the resistance of microorganisms to antimicrobials, which has a growing trend worldwide and poses a real danger in the treatment of infections. Microbial resistance was monitored in Salmonella spp., E. coli , Campylobacter spp., Staphylococcus aureus and  Enterococcus spp.

The comprehensive report, which will be published as a publication, has a length of more than 130 pages, will be published in printed form, as a publication with an assigned ISBN. Summaries of individual chapters will be translated into English and published in an electronic version as a publication with an assigned ISBN.

See the appendices for more information.

Attachments (downloadable documents)


Research – Desiccation survival in Salmonella enterica, Escherichia coli and Enterococcus faecium related to initial cell concentration and cellular components

Journal of Food Protection

Salmonella enterica is well-known for its ability to survive and persist in low-moisture environments.  Previous studies have indicated a link between the initial cell concentration and the population of Salmonella that survive upon desiccation and subsequent storage; however, how the initial cell concentration affects survival is unknown.  This study examined the basis of this phenomena and whether it occurred in other microorganisms, specifically Shiga toxigenic Escherichia coli (STEC), and Enterococcus faecium . Salmonella, STEC, and E. faecium were grown as lawns on TSAYE and harvested using buffered peptone water (BPW). To determine recovery at different initial cell levels, cultures were diluted to 9, 7, and 5 log CFU/mL and applied to filters.  Filters were dried for 24 h, then stored for 28 d at 25°C/33% RH. During storage, cells were recovered from filters using BPW and cultivated on TSAYE.  Both Salmonella and E. coli , but not E. faecium , showed non-proportional recovery. Less viability remained with lower initial starting population after 24 h desiccation such that ≥10 log CFU/mL were recovered when 11 log CFU/mL was desiccated, but ❤ log CFU/mL were recovered when 5 log CFU/mL was desiccated. Once dried, persistence did not appear affected by initial cell concentration. When dead cells (heat-treated) were added to the diluent, recovery of Salmonella was proportional with respect to the initial cell concentration. To further examine the response on desiccation, Salmonella was diluted in BPW containing one of 11 different test cell components related to quorum sensing or known to affect desiccation resistance to assess recovery and persistence. Of the 11 additions only cell debris fractions, cell-free extract, and peptidoglycan improved recovery of Salmonella . Desiccation survival appears related to cell wall components, however, the exact mechanism affecting survival remains unknown.

Research – Enterococci from Raw-Milk Cheeses: Current Knowledge on Safety, Technological, and Probiotic Concerns


The present study is focused on the safety, technological characteristics, and probiotic evaluation of Enterococcus species from different artisanal raw milk dairy products, mainly cheeses with ripening. Apart from proteolytic and lipolytic activities, most enterococci show the ability to metabolize citrate and convert it to various aromatic compounds. Long-ripened cheeses therefore have a specific flavor that makes them different from cheeses produced from thermally treated milk with commercial starter cultures. In addition, enterococci are producers of bacteriocins effective against spoilage and pathogenic bacteria, so they can be used as food preservatives. However, the use of enterococci in the dairy industry should be approached with caution. Although originating from food, enterococci strains may carry various virulence factors and antibiotic-resistance genes and can have many adverse effects on human health. Still, despite their controversial status, the use of enterococci in the food industry is not strictly regulated since the existence of these so-called desirable and undesirable traits in enterococci is a strain-dependent characteristic. To be specific, the results of many studies showed that there are some enterococci strains that are safe for use as starter cultures or as probiotics since they do not carry virulence factors and antibiotic-resistance genes. These strains even exhibit strong health-promoting effects such as stimulation of the immune response, anti-inflammatory activity, hypocholesterolemic action, and usefulness in prevention/treatment of some diseases. View Full-Text

Research- Predictive microbial modeling of E. faecium NRRL B-2354 inactivation during baking of a multi-component low-moisture food

Journal of Food Protection

Validating baking ovens as a microbial kill step, using thermal inactivation models, is desirable; however, traditional isothermal models may not be appropriate for these dynamic processes, yet they are being used by the food industry. Previous research indicates that the impact of additional process conditions, such as process humidity, should be considered when validating thermal processes for the control of microbial hazards in low-moisture foods. In this study, the predictive performance of traditional and modified thermal inactivation kinetic models accounting for process humidity were assessed for predicting bacterial inactivation of Enterococcus faecium NRRL B-2354 in a multi-ingredient composite food during baking. Ingredients (milk powder, protein powder, peanut butter, and whole wheat flour), individually inoculated to ~6 logCFU/g and equilibrated to a water activity of 0.25, were mixed to form a dough. An isothermal inactivation study was conducted for the dough to obtain traditional D- and z- values (n=63). In a separate experiment, cookies were baked under four dynamic heating conditions: 135℃/high humidity, 135℃/low humidity, 150℃/high humidity, and 150℃/low humidity. Process humidity measurements, time-temperature profiles for the product core, surface, and bulk air, and microbial survivor ratios were collected for the four conditions at six residence times (n=144). The traditional isothermal model had a poor root mean square error (RMSE) of 856.51 log (CFU/g), significantly overpredicting bacterial inactivation during the process. The modified model accounting for the dynamic time-temperature profile and process humidity data yielded a better predictive performance with a RMSE of 0.55 log CFU/g. The results demonstrate the importance of accounting for additional process parameters in baking inactivation models, and that model performance can be improved by utilizing model parameters obtained directly from industrial-scale experimental data.

Research – Thermal Inactivation Kinetics of Salmonella and Enterococcus faecium NRRL-B2354 on whole chia seeds (Salvia hispanica L.)

Journal of Food Protection

Intervention technologies for inactivating Salmonella in whole chia seeds are currently limited. The determination of the thermal inactivation kinetics of Salmonella o n chia seeds and selection of an appropriate nonpathogenic surrogate will provide a knowledge foundation for selecting and optimizing thermal pasteurization processes for chia seeds. In this study, chia seed samples from three separate production lots were inoculated with a five strain Salmonella cocktail or Enterococcus faecium NRRL-B2354 and equilibrated to 0.53 aw at room temperature (25 °C). After equilibration for at least three days, the inoculated seeds were subjected to isothermal treatments at 80, 85, or 90 °C. Samples were taken out at six timepoints and enumerated for survivors. Initial dilution of whole chia seeds was performed in a filter bag at a 1:30 ratio after it was shown to have similar recovery to grinding the seeds. Survivor data were fitted to consolidated models consisting of a primary model (log-linear or Weibull) and one secondary model (Bigelow). E. faecium exhibited higher thermal resistance than Salmonella , suggesting its suitability as a conservative nonpathogenic surrogate. The Weibull model was a better fit for the survivor data than the log-linear model for both bacteria due to its lower root mean square error and corrected Akaike’s Information Criterion values. Measurements of lipid oxidation and fatty acid content indicated a few statistically different values compared to the control samples, but the overall difference in magnitudes were relatively small. The thermal inactivation kinetics of Salmonella and E. faecium o n chia seeds as presented in this study can serve as a basis for developing thermal pasteurization processes for chia seeds.