Antilisterial Peptides Released by Enzymatic Hydrolysis from Grass Carp Proteins and Activity on Controlling L. monocytogenes Inoculated in Surimi Noodle

The primary objective of this study was to prepare the antilisterial peptides by enzymatic (pepsin, trypsin, protamex, neutrase, flavourzyme, papain, alcalase, and acid protease) hydrolysis of grass carp proteins with various degree of hydrolysis. The second objective was to evaluate the antilisterial activity of grass carp proteins hydrolysates (GCPH) at differnet levels in the surimi noodle samples with or without boiling treatment inoculated with 10⁴ CFU/g of Listeria monocytogenes for storage at 4 and 25 °C up to 20 d. These results revealed that GCPH, obtained by treatment with the neutrase hydrolysates at degree of hydrolysis of 19% (NSH19), showed the highest antilisterial activity reaching up to 64.8% inhibition of bacterial growth. The antilisterial activity of NSH19 in the raw surimi noodle was stronger than that of the boiled group, and the samples treated by NSH19 at 10 g/100 g level had no‐detectable numbers of L. monocytogenes for both raw and boiled noodle samples within 20 d of storage at 4 and 25 °C. The results of this study indicated that antilisterial peptides generated via neutrase from grass carp proteins can efficiently inhibit the growth of L. monocytogenes in surimi noodle, which was useful as natural preservatives for storage and distribution of meat based products.     

Effect of partial replacement of potassium lactate and sodium diacetate by natural green tea and grape seed extracts and postpackaging thermal treatment on the growth of Listeria monocytogenes in hotdog model system

Low‐ (5%) and high‐fat (20%) chicken and turkey hotdogs were formulated in three groups: no antimicrobials (control), chemical preservatives (potassium lactate and sodium diacetate) alone and partial replacement of chemical preservatives by green tea (GTE) and grape seed extracts (GSE), surface inoculated (c. 10³ CFU g^?1) with Listeria monocytogenes, treated with or without heat treatment (65 °C for 104 s) to determine the growth of L. monocytogenes until spoilage (28 days). Maximum growth inhibitions (c. 2.0 CFU g^?1) were observed in the treatments having chemical preservatives and plant extracts regardless of the meat and fat type. Furthermore, plant extracts along with chemical preservatives demonstrated additional inhibitory effect on the growth of L. monocytogenes survivors in chicken hotdog samples followed by postpackaging heat treatment. Results demonstrated that natural GTE and GSE can partially replace the chemical preservatives and further enhance the antilisterial activities when combined with heat treatment.     

Effects of curing sodium nitrite additive and natural meat fat on growth control of Listeria monocytogenes by the bacteriocin-producing Lactobacillus curvatus strain CWBI-B28

In realistic model meat systems, the separate and combined effects of fat content and sodium nitrite on the antilisterial activity of the bacteriocin of Lactobacillus curvatus CWBI-B28 were studied. In laboratory fermentations where Listeria monocytogenes was co-cultured at 4 °C with bacteriocin-producing CWBI-B28 in lean pork meat (fat content: 13%) without added nitrite, a strong antilisterial effect was observed after one week. The effect was maintained for an additional week, after which a slight and very gradual rebound was observed. Both added nitrite (20 ppm) and a high-fat content (43%) were found to antagonise this antilisterial effect, the Listeria cfu count reached after six weeks being 200 times as high in high-fat meat with added nitrite than in lean meat without nitrite. This antagonism could not be attributed to slower growth of the bacteriocin-producing strain, since CWBI-B28 grew optimally in fat-rich meat with 20 ppm sodium nitrite. Bacteriocin activity was also measured in the samples. The observed activity levels are discussed in relation to the degree of antilisterial protection conferred.     

Antioxidant and antimicrobial activity and potential of heather (Erica spp.) extracts in the control of Listeria monocytogenes

In this work, heather and its flowers were studied regarding their antioxidant and antimicrobial activities. Plants were subjected to ultrasound-assisted methanolic extraction followed by fractionation. A phytochemical characterisation of extracts content in total phenols and flavonoids, and their antioxidant activity was performed. The antimicrobial activity was evaluated through the determination of minimum inhibitory concentration and by bioautography. Following, studies on the antilisterial potential were carried out by: time-kill curves, inhibition of biofilm formation and tolerance of Listeria monocytogenes to adverse conditions. The results evidenced the antioxidant activity in both extracts, as well as, the antimicrobial activity against Gram-positive bacteria. Concerning the evaluation of the antilisterial potential, a bacteriostatic behaviour and inhibition of biofilms formation ability were observed. Listeria monocytogenes showed an increased susceptibility to adverse conditions when preincubated with extracts. Thus, heather and its flowers may be a source of new compounds with antilisterial activity potential.     

Antimicrobial activity of lactic acid bacteria against Listeria monocytogenes on frankfurters formulated with and without lactate/diacetate

Contamination by Listeria monocytogenes has been a constant public health threat for the ready-to-eat (RTE) meat industry due to the potential for high mortalities from listeriosis. Lactic acid bacteria (LAB) have shown protective action against various pathogenic bacteria. The aim of this study was to evaluate the antilisterial activity of a combination of three LAB strains (Lactiguard®) on L. monocytogenes. The combination of the LAB was inhibitory to L. monocytogenes inoculated onto frankfurters not containing lactate/diacetate after 8 weeks of refrigerated storage (0.6 log reduction compared to L. monocytogenes only control), and when a cell free extract (CFS) of the LAB was added with LAB even more inhibition was obtained (1.2 log reduction compared with L. monocytogenes only). In frankfurters containing lactate/diacetate the LAB and the LAB plus CFS were more effective in reducing growth of L. monocytogenes after 8 weeks of refrigerated storage (2 and 3.3 log reductions respectively).     

Antimicrobial activity of lactic acid bacteria against Listeria monocytogenes on frankfurters formulated with and without lactate/diacetate

Contamination by Listeria monocytogenes has been a constant public health threat for the ready-to-eat (RTE) meat industry due to the potential for high mortalities from listeriosis. Lactic acid bacteria (LAB) have shown protective action against various pathogenic bacteria. The aim of this study was to evaluate the antilisterial activity of a combination of three LAB strains (Lactiguard®) on L. monocytogenes. The combination of the LAB was inhibitory to L. monocytogenes inoculated onto frankfurters not containing lactate/diacetate after 8 weeks of refrigerated storage (0.6 log reduction compared to L. monocytogenes only control), and when a cell free extract (CFS) of the LAB was added with LAB even more inhibition was obtained (1.2 log reduction compared with L. monocytogenes only). In frankfurters containing lactate/diacetate the LAB and the LAB plus CFS were more effective in reducing growth of L. monocytogenes after 8 weeks of refrigerated storage (2 and 3.3 log reductions respectively).     

Development of a New Paenibacillin‐Producing Strain and Testing its Usability in Improving Food Safety

A new bacterial strain that produces a bacteriocin (paenibacillin) without polymyxin was developed from Paenibacillus polymyxa that co‐produces the 2 antimicrobial agents. Gamma radiation was used successfully to develop the new strain, P. polymyxa OSY‐HG. Subsequently, we explored the feasibility of using food or food ingredients as growth media for the new strain. Milk supported the growth of P. polymyxa OSY‐HG which produced up to 32 mg paenibacillin/L milk without polymyxin. Fermentation crude extract was applied in a model food (Vienna sausage) to control Listeria innocua, a Listeria monocytogenes surrogate. The treatment increased Listeria lag time by 2 d at 7 °C and at least 6 h at 37 °C. In conclusion, a new paenibacillin‐producing P. polymyxa strain has been developed for potential industrial use. Using the new strain in applications that enhance food safety is feasible.     

Combined use of bacteriocin‐producing strains to control Listeria monocytogenes regrowth in raw pork meat

Avoiding the presence of Listeria in meat and dairy products is a major challenge for the food industry. In this work, a Lactobacillus curvatus strain producing the bacteriocin sakacin P and a Pediococcus acidilactici strain producing another bacteriocin, pediocin AcH, were used as starter cultures under laboratory conditions in a Listeria‐seeded raw‐pork‐meat matrix, which was then stored for 6 weeks at 4 °C. At 1 week intervals during the storage period, the antilisterial activity was evaluated. When either strain was added alone, the Listeria monocytogenes cfu count initially dropped from 10² cfu g^?1 to an undetectable level by the end of week 1 or 2, but this was followed by a rebound (regrowth) 1 week later. When both strains were added together to the meat matrix, rebound was delayed, Listeria remaining undetected from the end of week 1 to the end of week 5. A rebound was observed 6 weeks post‐inoculation, but fewer than 10 cfu g^?1 were counted. The use of more than one bacteriocin‐producing strain may thus overcome some of the problems limiting the effectiveness of bacteriocins in food systems.     

Listeria monocytogenes in Vacuum‐Packed Smoked Fish Products: Occurrence,Routes of Contamination,and Potential Intervention Measures

The occurrence of Listeria monocytogenes in ready‐to‐eat (RTE) fish products is well documented and represents an important food safety concern. Contamination of this pathogen in vacuum‐packed (VP) smoked fish products at levels greater than the RTE food limit (100 CFU/g) has been traced to factors such as poor sanitary practices, contaminated processing environments, and temperature abuse during prolonged storage in retail outlets. Intervention technologies including physical, biological, and chemical techniques have been studied to control transmission of L. monocytogenes to these products. High‐pressure processing, irradiation, and pulsed UV‐light treatment have shown promising results. Potential antilisterial effects of some sanitizers and combined chemical preservatives have also been demonstrated. Moreover, the concept of biopreservation, use of bioactive packaging, and a combination of different intervention technologies, as in the hurdle concept, are also under consideration. In this review, the prevalence, routes of contamination, and potential intervention technologies to control transmission of L. monocytogenes in VP smoked fish products are discussed.     

Influence of composition and processing of Frankfurter sausages on the growth dynamics of Listeria monocytogenes under vacuum

Ready-to-eat (RTE) meat products, such as Frankfurter sausages, are often linked to cases of listeriosis, which is a disease caused by the foodborne pathogen Listeria monocytogenes. Frankfurter sausages vary, from manufacturer to manufacturer, in many aspects: (i) composition, (ii) physicochemical characteristics, (iii) background flora, (iv) processing technology and (v) organoleptic characteristics. Some of these factors have been widely investigated for their effect on L. monocytogenes on food products. However, studies on the specific effect of composition and processing technology on L. monocytogenes dynamics between two sausages are lacking. In this study, the growth dynamics of L. monocytogenes on two types of Frankfurter sausages, fresh and in brine, were studied at constant storage temperatures (4, 8 and 12 °C). Additionally, the physicochemical and compositional characteristics of both types of sausages were analysed. In order to study the isolated effect of preservatives, L. monocytogenes growth dynamics, at 4 °C and 30 °C, were studied in two types of liquid systems. These were prepared with the same level of preservatives as in the two types of sausages. Results indicated no major significant differences in physicochemical characteristics for the two types of sausages; but, statistically significant variability was detected in the concentration of preservatives. In liquid systems, the maximum specific growth rate (μ_max) remained unaffected by the effect of preservatives, but the lag phase was longer in the system mimicking fresh sausages. In sausages, the ‘in brine’ type had two-fold higher μ_max at all temperatures and shorter lag phase at 4 °C. The presence/absence of sausage skin, which was found to be impermeable from L. monocytogenes cells and was present in the fresh sausage, could explain those differences. In conclusion, this study highlighted the influence of processing factors, and specifically of the sausage casing on L. monocytogenes growth dynamics. Therefore, an edible membrane, which is heat resistant and impermeable to the cells, could be a hurdle strategy to control the microbiological food safety.     

Microbial and Physicochemical Characterization of the Horse Meat in Fermented Sausage

In this study, 150 lactic acid bacterial strains, originating from horse meat, were subjected to a step-by-step screening and characterization to search for potential protective cultures to be used in the meat industry. Strains were first tested for their homofermentation and salt tolerance. Second, the antibacterial capacities towards Listeria monocytogenes were determined in an agar spot test. In total, 50% of the tested strains were inhibitory towards Listeria monocytogenes. However, only 12 strains produced a bacteriocin. Finally, three strains with strong bacteriocin activity were competitively evaluated by comparing their growth rate, acidifying character and lactic acid production at 15°C under anaerobic conditions in a liquid broth. All three strains had a fast growth rate with rapid acidification caused by the production of high levels of lactic acid. Based on this Lactobacillus sakei was used as starter culture for producing sausage horse meat. In this study, fermentations were followed analyzing the microbiological and physicochemical aspects of this product. The sausages were characterized based on microbial activity of lactic acid bacteria that resulted in a product with a final pH of about 4.56. No Listeria monocytogenes, Salmonella spp. nor Sulfite reducing clostridia were ever isolated from the raw materials or the fermented sausages during the maturation, reflecting the safety of this product. The final water activity of the product was 0.85. Lactobacillus sakei was efficient in reducing amine production since this strain caused a quick pH drop during sausage fermentation.     

Hot topic: Antilisterial activity by endolysin PlyP100 in fresh cheese

Our objective was to assess the antimicrobial efficacy of a Listeria bacteriophage endolysin that may address limitations of current antilisterial processes for fresh cheeses. Listeria monocytogenes is highly problematic in the manufacture and processing of ready-to-eat foods due to its environmental persistence and its ability to grow under refrigerated storage. Special care must be taken to prevent listerial contamination during the production of fresh cheeses, as their delicate flavor and texture are incompatible with many of the antimicrobial processes and additives commonly used for other foods. Bacteriophage-derived cell wall hydrolytic enzymes, known as endolysins, comprise one possible intervention that may not suffer from the high strain specificity of their parent bacteriophages or the development of resistant strains. We recombinantly expressed endolysin PlyP100 and compared its lytic activity in vitro across several environmental parameters and target organisms, then incorporated it into a fresh cheese model challenged with a cocktail of L. monocytogenes. We show that PlyP100 demonstrates optimal activity under pH and salt concentrations consistent with a low-acid food matrix such as fresh cheese. Furthermore, we show that PlyP100 exhibits target specificity for gram-positive organisms with directly crosslinked peptidoglycan and displays considerable inhibitory activity against L. monocytogenes in fresh cheese for at least 4 wk under refrigerated storage. As PlyP100 demonstrates considerable promise for preventing the propagation of L. monocytogenes in fresh cheeses, this novel preservation method could help safeguard consumer health and the market expansion of an otherwise high-risk food with few other viable preservatives.     

Efficacy of freezing, frozen storage and edible antimicrobial coatings used in combination for control of Listeria monocytogenes on roasted turkey stored at chiller temperatures

The presence and growth of Listeria monocytogenes on ready-to-eat (RTE) turkey is an important food safety issue. The antilisterial efficacy of four polysaccharide-based edible coatings (starch, chitosan, alginate and pectin) incorporating sodium lactate (SL) and sodium diacetate (SD) as well as commercial preparations Opti.Form PD4, NovaGARD™ CB1, Protect-M and Guardian™ NR100 were compared against L. monocytogenes on roasted turkey. Pectin coating treatments incorporating SL/SD, Opti.Form PD4 with or without Protect-M, and NovaGARD™ CB1 displayed higher antimicrobial efficacy against.L. monocytogenes than the other antimicrobials and coating materials. In the second phase of the study, it was investigated whether frozen storage could enhance the antilisterial effectiveness of pectin coating treatments on chilled roasted turkey. Inoculated roasted turkey samples coated with pectin-based treatments were frozen for up to 4 weeks and subsequently stored at 4 °C for 8 weeks. Frozen storage significantly enhanced the antilisterial activity of various coating treatments; with selected treatments reducing the L. monocytogenes populations by as much as 1.1 log CFU/cm² during the subsequent 8-week chilled storage. This study demonstrates that pectin-based antimicrobial edible coatings hold promise in enhancing the safety of RTE poultry products and frozen storage has the potential to enhance their effectiveness.     

Enhancement of antilisterial activity of essential oil constituents by nisin and diglycerol fatty acid ester

Plant-derived essential oil components in combination with nisin and diglycerol fatty acid esters were investigated for their antibacterial activity against Listeria monocytogenes. Carvacrol and thymol were found to have the strongest antilisterial properties, followed by eugenol, cinnamaldehyde and isoeugenol. The antilisterial activity of the other essential oils (limonene, pinene, ally lisothiocyanate and linalool) was found to be low, even at the highest concentration used (0.1%). Among the diglycerol esters of fatty acids with different carbon chain lengths, diglycerol monolaurate was the most active against L. monocytogenes. A combined antilisterial effect was observed between nisin and the essential oils (carvacrol, thymol and eugenol); moreover, the addition of diglycerol monolaurate as a third preservative factor led to further combined antilisterial activities between the essential oil constituents (carvacrol, thymol and eugenol) and nisin even at lower, sub-lethal concentrations. These results indicate that nisin and diglycerol monolaurate can be used to enhance the antilisterial activity of essential oils, allowing for a reduction in the dosage used in food preservation and thereby resulting in the reduction of undesirable flavors.     

Inhibitory Activity of Paenibacillus polymyxa on the Biofilm Formation of Cronobacter spp. on Stainless Steel Surfaces

The objective of this study was to control the survival or biofilm formation of Cronobacter spp. on stainless steel surfaces using Paenibacillus polymyxa. The antibacterial activity of a cell‐free culture supernatant (CFCS) of P. polymyxa against Cronobacter spp. was found to vary with P. polymyxa incubation time. Maximum activity occurred when P. polymyxa was incubated at 25 or 30 °C for 96 h. When the CFCS was introduced to Cronobacter spp. adhered to stainless steel strips at 25 °C for up to 72 h, the CFCS successfully inhibited Cronobacter biofilm formation. Additionally, stainless steel surfaces with a preformed P. polymyxa biofilm were exposed to Cronobacter spp. suspensions in PBS or 0.1% peptone water at 3, 5, or 7 log CFU/mL to facilitate its attachment. The Cronobacter population significantly decreased on this surface, regardless of inoculum level or carrier, when the P. polymyxa biofilm was present. However, the microbial population decreased within 6 h and remained unchanged thereafter when the surface was immersed in an inoculum suspended in 0.1% peptone water at 5 or 7 log CFU/mL. These results indicate that P. polymyxa is able to use a promising candidate competitive‐exclusion microorganism to control Cronobacter spp.     

Antilisterial Activity of Hops Beta Acids in Broth with or Without Other Antimicrobials

ABSTRACT: Hops beta acids (HBA) are parts of hops flowers used to preserve wort and provide flavor in beer, and are reported as having antimicrobial properties. This study evaluated the antilisterial activity of HBA alone or in combination with other known antimicrobials in a culture broth medium. Listeria monocytogenes (10‐strain mixture) was inoculated (2.6 to 2.8 log CFU/mL) into tryptic soy broth supplemented with 0.6% yeast extract (TSBYE) without (control) or with HBA (0.5 to 5.0 μg/mL), potassium lactate (1.0%), sodium diacetate (0.25%), or acetic acid (0.1%), alone or in combination with HBA (0.5 to 3.0 μg/mL). Survival/growth of the pathogen during storage at 4 °C (35 d), 10 °C (20 d), or 25 °C (2 d) was periodically monitored by spiral plating onto tryptic soy agar plus 0.6% yeast extract. As expected, TSBYE without antimicrobials (control) supported rapid pathogen growth with growth rates of 0.40, 2.88, and 9.58 log CFU/mL/d at 4, 10, and 25 °C, respectively; corresponding Y_end values exceeded 9.0 log CFU/mL at 35, 20, and 2 d storage. HBA used alone (1.0 to 5.0 μg/mL) inhibited growth of L. monocytogenes at all 3 temperatures, with inhibition being more pronounced at higher concentrations and at the lower storage temperature (4 °C). The antilisterial activity of HBA (0.5 to 3.0 μg/mL) was enhanced when combined with sodium diacetate, acetic acid, or potassium lactate, achieving complete inhibition at 4 °C when 3.0 μg/mL HBA were used in combination with each of the above antimicrobials. Overall, HBA exhibited promising antilisterial activity in a broth medium and further studies are needed to investigate its potential antilisterial effects in food products.     

Control of Listeria monocytogenes Contamination in Ready‐to‐Eat Meat Products

The ubiquitous nature of Listeria monocytogenes and its ability to grow at refrigerated temperature makes L. monocytogenes a significant threat to the safety of ready‐to‐eat (RTE) meat products. The contamination by L. monocytogenes in RTE meat primarily occurs during slicing and packaging after cooking. The effectiveness of post‐package decontamination technology such as in‐package thermal pasteurization, irradiation, and high‐pressure processing are discussed. Formulating meat products with antimicrobial additives is another common approach to control L. monocytogenes in RTE meat. Irradiation is an effective technology to eliminate L. monocytogenes but can influence the quality of RTE meat products significantly. The effect of irradiation or the combination of irradiation and antimicrobials on the survival of L. monocytogenes and the quality of RTE meat is discussed.     

Functional properties of novel protective lactic acid bacteria and application in raw chicken meat against Listeria monocytogenes and Salmonella enteritidis

In this study 635 lactic acid bacteria of food origin were evaluated for their potential application as protective cultures in foods. A stepwise selection method was used to obtain the most appropriate strains for application as protective cultures in chicken meat. Specifically, all strains were examined for antimicrobial activity against various Gram positive and Gram negative pathogenic and spoilage bacteria. Strains exhibiting anti-bacterial activity were subsequently examined for survival in simulated food processing and gastrointestinal tract conditions, such as high temperatures, low pH, starvation and the presence of NaCl and bile salts. Selected strains where then examined for basic safety properties such as antibiotic resistance and haemolytic potential, while their antimicrobial activity was further investigated by PCR screening for possession of known bacteriocin genes. Two chosen strains were then applied on raw chicken meat to evaluate their protective ability against two common food pathogens, Listeria monocytogenes and Salmonella enteritidis, but also to identify potential spoilage effects by the application of the protective cultures on the food matrix. Antimicrobial activity in vitro was evident against Gram positive indicators, mainly Listeria and Brochothrix spp., while no antibacterial activity was obtained against any of the Gram negative bacteria tested. The antimicrobial activity was of a proteinaceous nature while strains with anti-listerial activity were found to possess one or more bacteriocin genes, mainly enterocins. Strains generally exhibited sensitivity to pH 2.0, but good survival at 45 °C, in the presence of bile salts and NaCl as well as during starvation, while variable survival rates were obtained at 55 °C. None of the strains was found to be haemolytic while variable antibiotic resistance profiles were obtained. Finally, when the selected strains Enterococcus faecium PCD71 and Lactobacillus fermentum ACA-DC179 were applied as protective cultures in chicken meat against L. monocytogenes and S. enteritidis respectively, a significantly reduced growth of these pathogenic bacteria was observed. In addition, these two strains did not appear to have any detrimental effect on biochemical parameters related to spoilage of the chicken meat.     

Synergistic effect of nisin and garlic shoot juice against Listeria monocytogenes in milk

The aim of this research was to determine the synergistic effect of nisin and garlic shoot juice (GSJ) against Listeria monocytogenes ATCC 19118 found in whole (3.5%), low (1%) and skim (no fat content) milk. Garlic shoot juice (GSJ) at concentrations of 2.5%, 5% and 10% revealed strong and similar patterns of antilisterial effect against L. monocytogenes ATCC 19118 in all categories of milk. Nisin only at concentrations of 62.5, 125, 250 and 500 IU/ml displayed a strong antilisterial effect as compared to the control group. Also, the synergistic combinations of GSJ (2.5%, 5%) and nisin (62.5, 125, 250 and 500 IU/ml) had a remarkable antilisterial activity in all categories of whole, low and skim milk after 14 days. Results of this study indicated the synergistic effect of GSJ and nisin as a potential antilisterial agent for the food industry.     

Survival and Metabolic Activity of Listeria monocytogenes on Ready‐to‐Eat Roast Beef Stored at 4 °C

Three brands of commercial roast beef were purchased and artificially inoculated with a 5‐strain Listeria monocytogenes cocktail at 2 inoculation levels (approximately 3 and 6 Log CFU/g). Although all 3 brands contained sodium diacetate and sodium lactate, inoculated Listeria cocktail survived for 16 d in all 3 brands; significant increases in L. monocytogenes numbers were seen on inoculated Brand B roast beef on days 12 and 16. Numbers of L. monocytogenes increased to 4.14 Log CFU/g for the 3 Log CFU/g inoculation level and increased to 7.99 Log CFU/g for the 6 Log CFU/g inoculation level by day 16, with the pH values being 5.4 and 5.8 respectively. To measure the cell viability in potential biofilms formed, an Alamar blue assay was conducted. Brand B meat homogenate had the highest metabolic activities (P < 0.05). By comparing its metabolic activities to Brands A and C and the inoculated autoclaved meat homogenates, results indicated that the microflora present in Brand B may be the reason for high metabolic activities. Based on the denaturing gradient gel electrophoresis and the Shannon–Wiener diversity index analysis, the “Brand” factor significantly impacted the diversity index (P = 0.012) and Brand B had the highest microflora diversity (Shannon index 1.636 ± 0.011). Based on this study, results showed that antimicrobials cannot completely inhibit the growth of L. monocytogenes in ready‐to‐eat roast beef. Native microflora (both diversity and abundance), together with product formula, pH, antimicrobial concentrations, and storage conditions may all impact the survival and growth of L. monocytogenes.