Inactivation of Vibrio parahaemolyticus in Hard Clams (Mercanaria mercanaria) by High Hydrostatic Pressure (HHP) and the Effect of HHP on the Physical Characteristics of Hard Clam Meat

Shellfish may internalize dangerous pathogens during filter feeding. Traditional methods of depuration have been found ineffective against certain pathogens. The objective was to explore high hydrostatic pressure (HHP) as an alternative to the traditional depuration process. The effect of HHP on the survival of Vibrio parahaemolyticus in live clams (Mercanaria mercanaria) and the impact of HHP on physical characteristics of clam meat were investigated. Clams were inoculated with up to 7 log CFU/g of a cocktail of V. parahaemolyticus strains via filter feeding. Clams were processed at pressures ranging from 250 to 552 MPa for hold times ranging between 2 and 6 min. Processing conditions of 450 MPa for 4 min and 350 MPa for 6 min reduced the initial concentration of V. parahaemolyticus to a nondetectable level (<10¹ CFU/g), achieving >5 log reductions. The volume of clam meat (processed in shell) increased with negligible change in mass after exposure to pressure at 552 MPa for 3 min, while the drip loss was reduced. Clams processed at 552 MPa were softer compared to those processed at 276 MPa. However, all HHP processed clams were found to be harder compared to unprocessed. The lightness (L*) of the meat increased although the redness (a*) decreased with increasing pressure. Although high pressure‐processed clams may pose a significantly lower risk from V. parahaemolyticus, the effect of the accompanied physical changes on the consumer's decision to purchase HHP clams remains to be determined. Practical Application : Shellfish may contain dangerous foodborne pathogens. Traditional methods of removing those pathogen have been found ineffective against certain pathogens. The objective of this research was to determine the effect of high hydrostatic pressure on V. parahaemolyticus in clams. Processing conditions of 450 MPa for 4 min and 350 MPa for 6 min reduced the initial concentration of V. parahaemolyticus to a nondetectable level, achieving >5 log reductions.     

Inactivation of Vibrio parahaemolyticus and Vibrio vulnificus in oysters by high-hydrostatic pressure and mild heat

Several recent outbreaks associated with oysters have heightened safety concerns of raw shellfish consumptions, with the majority being attributed to Vibrio spp. The objective of this study was to determine the effect of high-hydrostatic pressure (HHP) followed by mild heating on the inactivation of Vibrio parahaemolyticus and Vibrio vulnificus in live oysters. Inoculated oysters were randomly subjected to: a) pressurization at 200–300 MPa for 2 min at 21 °C, b) mild heat treatment at 40, 45 or 50 °C for up to 20 min and c) pressure treatment of 200–300 MPa for 2 min at 21 °C followed by heat treatment at 40–50 °C. Counts of V. parahaemolyticus and V. vulnificus were then determined using the most probable number (MPN) method. Pressurization at 200–300 MPa for 2 min resulted in various degrees of inactivation, from 1.2 to >7 log MPN/g reductions. Heat treatment at 40 and 45 °C for 20 min only reduced V. parahaemolyticus and V. vulnificus by 0.7–2.5 log MPN/g while at 50 °C for 15 min achieved >7 log MPN/g reduction. HHP and mild heat had synergistic effects. Combinations such as HHP at 250 MPa for 2 min followed by heat treatment at 45 °C for 15 min and HHP at 200 MPa for 2 min followed by heat treatment at 50 °C for 5 min reduced both V. parahaemolyticus and V. vulnificus to non-detectable levels by the MPN method (<3 MPN/g). HHP at ≥275 MPa for 2 min followed by heat treatment at 45 °C for 20 min and HHP at ≥200 MPa for 2 min followed by heat treatment at 50 °C for 15 min completely eliminated both pathogens in oysters (negative enrichment results). This study demonstrated the efficiency of HHP followed by mild heat treatments on inactivation of V. parahaemolyticus and V. vulnificus and could help the industry to establish parameters for processing oysters.     

Behavior and incidence of Vibro parahaemolyticus in Sydney rock oysters (Crassostrea commercialis)

Vibrio parahaemolyticus grew poorly or not at all during storage of unopened Sydney rock oysters (Crassostrea commercialis) at 15 and 30°C for 2 and 7 days. Although V. parahaemolyticus counts often increased at 30°C, counts above 10⁴/g were not observed. Escherichia coli counts did not usually change substantially under these conditions. V. parahaemolyticus grew more readily during storage of unopened oysters under more severe conditions, with counts approaching or exceeding 10⁶/g after continuous or intermittent storage at 37°C. Opened oysters provided a much more favourable environment than unopened oysters for growth of V. parahaemolyticus. Growth occurred at 15, 30 and 37°C, with counts > 10⁶/g after overnight storage at 30 or 37°C. A survey of 30 market samples of oysters was conducted. Sixteen samples of unopened oysters were collected at the wholesale level and 14 samples of refrigerated opened oysters were purchased from retailers. V. parahaemolyticus was present in all samples of unopened oysters (range 0.4/g?2.3 × 10⁴/g) and in 13 samples of opened oysters (range 4.3/g? > 1.1 × 10³/g).     

Effect of initial aflatoxin concentration,heating time and roasting temperature on aflatoxin reduction in contaminated peanuts and process optimisation using response surface modelling

Response surface methodology was applied to optimise the aflatoxin reduction in both naturally and artificially contaminated samples using dry oven. The effect of initial aflatoxin concentration (0–400 ng g^?1), heating time (30–120 min) and temperature (90–150 °C) was evaluated. The maximum reduction of AFB1 (78.4%) and AFB2 (57.3%) of artificially contaminated samples with initial aflatoxin concentration of 237 and 68 ng g^?1, and those of AFG1 (73.9%) and AFG2 (75.2%) with initial aflatoxin concentration of 215 and 75 ng g^?1 was obtained at 150 °C. The maximum reduction of AFB1 (80.2%) and AFB2 (69.7%) of naturally contaminated samples with initial aflatoxin concentration of 174 and 25 ng g^?1 was obtained at 150 °C and 130 °C, respectively.     

Microbiological quality of retail fresh fish fillets in The Netherlands

A total of 242 samples of ready-for-sale fish fillets of validated good sensory quality was examined for colony counts at 20, 30 and 37°C, Enterobacteriaceae at 37°C, Escherichia coli, Salmonella and Vibrio parahaemolyticus in 10 g aliquots. Staphylococcus aureus and yeast and mould propagules. Gram negative pathogens were not detected in any sample. The following reference values were found attainable: colony counts at 30°C, 10⁶ g^?1; E. coli 10 g^?1; S. aereus 10² g^?1; yeast and mould propagules 10⁴ g^?1. These reference values include, as customary, a tolerance of about 20% of samples exceeding the stated levels without, however, reaching the next log₁₀ level.     

Microbiological hazards involved in fresh‐cut lettuce processing

BACKGROUND: The increasing consumption of produce all over the world has resulted in increasing concern by the regulatory agencies with respect to the level of safety performed by the processors. The objective of the present study was to investigate the hazards involved in the various steps of fresh‐cut lettuce processing (reception/selection of raw material, washing, rinsing, sanitisation and final product) by means of microbiological analyses of microbial groups used as indicators of hygienic conditions and of pathogens. RESULTS: High microbial loads of mesophilic and psychrotrophic bacteria and Pseudomonas spp. were found in the ram reception (～6 log colony‐forming units (CFU) g^?1), which were reduced by a single logarithmic cycle for the last two microbial groups after the sanitisation step (P < 0.05), the latter being ineffective against the first microbial group (P > 0.05). Lower counts of yeasts and moulds, total coliforms (35 °C) and faecal coliforms (44 °C) were observed in the initial step (3.49–4.53 log CFU g^?1, 0.65–1.55 log most probable number (MPN) g^?1 and 0.50–0.90 log MPN g^?1 respectively), these values increasing significantly after the sanitisation step for yeasts and moulds (～5 log CFU g^?1) but remaining unaltered for coliforms (P > 0.05). Salmonella spp. were not found in any of the experiments carried out, while the presence of Escherichia coli was observed in the final product. CONCLUSIONS : Practices compromising the hygienic quality of the final product during commercial storage were observed and corrective measures suggested. To the best of the authors' knowledge, these are the first data on microbiological safety in Brazilian fresh‐cut processing plants. Copyright © 2008 Society of Chemical Industry     

Prevalence of Vibrio parahaemolyticus in seafoods and their processing environments as detected by duplex PCR

A duplex polymerase chain reaction (PCR) procedure targeting the genes gyrB and tl was established for specific identification of Vibrio parahaemolyticus from seafoods and processing environments. It could detect as few as 2.5 × 10² colony‐forming units mL^?1 in pure cultures. Direct detection of V. parahaemolyticus was also possible from artificially contaminated shrimp samples if combined with proteinase treatment. The homogeneous colonies on thiosulfate/citrate/bile salts/sucrose agar suspected to be V. parahaemolyticus or closely related species (n = 37) out of 259 samples of seafoods and processing environments were identified using the conventional method and duplex PCR. Both methods identified 17 isolates as V. parahaemolyticus from among the suspected isolates. Only one of the 17 V. parahaemolyticus isolates possessed the thermostable direct haemolysin gene (tdh) fragment as detected by different primer pairs in single PCR. Copyright © 2006 Society of Chemical Industry     

Response of Listeria monocytogenes and Vibrio parahaemolyticus to High Hydrostatic Pressure

Listeria monocytogenes Scott A and CA, were subjected at 23°C to hydrostatic pressures ranging from 2,380 to 3,400 atm and Vibrio parahaemolyticus T-3765-1 from 680 to 1,700 atm. For L. monocytogenes Scott A, pressurization in ultra-high temperature-processed (UHT) milk and raw milk appeared to provide a protective effect and lessened cell death as compared to pressurization in phosphate-buffered saline (100 mM, pH 7.0). A population of about 10⁶ CFU/mL L. monocytogenes was killed by exposure to 3,400 atm within 80 min at 23°C in UHT milk. A population of about 10⁶ CFU/mL V. parahaemolyticus was killed by exposure to 1,700 atm within 10 min at 23°C in clam juice.     

Reductions of Vibrio parahaemolyticus in Pacific oysters (Crassostrea gigas) by depuration at various temperatures

Consumption of raw oysters has been linked to several outbreaks of Vibrio parahaemolyticus infection in the United States. This study investigated effects of ice storage and UV-sterilized seawater depuration at various temperatures on reducing V. parahaemolyticus in oysters. Raw Pacific oysters (Crassostrea gigas) were inoculated with a mixed culture of five clinical strains of V. parahaemolyticus (10290, 10292, 10293, BE 98-2029 and 027-1c1) at levels of 10⁴⁻⁶ MPN/g. Inoculated oysters were either stored in ice or depurated in recirculating artificial seawater at 2, 3, 7, 10, 12.5, and 15 °C for 4–6 days. Holding oysters in ice or depuration of oysters in recirculating seawater at 2 or 3 °C for 4 days did not result in significant reductions (P > 0.05) of V. parahaemolyticus in the oysters. However, depuration at temperatures between 7 and 15 °C reduced V. parahaemolyticus populations in oysters by >3.0 log MPN/g after 5 days with no loss of oysters. Depuration at refrigerated temperatures (7–15 °C) can be applied as a post-harvest treatment for reducing V. parahaemolyticus in Pacific oysters.     

A new process for preparing transparent alkalised duck egg and its quality

When fresh duck (Anas plotyrhyncus) eggs (pH 8·0–8·5) are heated, their albumen develops a turbid gel. Through appropriate alkalisation (pH 11·5–12·8), the gel's transparency can be increased. The transparency of the heated duck egg-white is affected by pH value, heating temperature, heating rate and salt concentration. This research deals with the process for preparing the transparent alkalised duck egg and the change in its quality when stored. If fresh duck eggs are pickled in a solution of 42 g NaOH+50 g NaCl litre^?1 (25·3°C) for 8 days, removed, put in a water bath and heated at 70°C for 10 min they become transparent, their hardness and penetration increasing with storage. Total bacterial count and volatile basic nitrogen also increase with storage. The total bacterial count and the volatile basic nitrogen were 4·6 × 10⁶ cfu g^?1, 0·32 mg g^?1 when stored at a temperature of under 25°C for 4 weeks, respectively.     

Evaluation of a Double Layer Agar Plate For Direct Enumeration of Vibrio parahaemolyticus

A double layer agar plate (DLAP) was developed according to the thin agar layer (TAL) method as a 1‐step procedure for direct enumeration of injured Vibrio parahaemolyticus cells based on the formation of unique purple colonies by V. parahaemolyticus. The DLAP was prepared by overlaying an equal volume (10 mL) of a nonselective medium (tryptic soy agar supplemented with 1.5% NaCl) onto a selective medium (Bio‐Chrome Vibrio medium). The DLAP was capable of detecting V. parahaemolyticus in mixed cultures containing non‐Vibrio bacteria. Production of purple colonies by V. parahaemolyticus on DLAP was not affected by the growth of other bacteria, even when V. parahaemolyticus was only a small fraction (5%) of the entire bacterial population. Direct plating on DLAP was found as effective as the most probable number (MPN) method for recovering heat‐and cold‐injured V. parahaemolyticus cells, which could not be detected by direct plating on Bio‐Chrome Vibrio medium or thiosulfate‐citrate‐bile salts‐sucrose agar. The DLAP offers an alternative to the MPN method for detecting injured V. parahaemolyticus cells and can be used as a simple 1‐step procedure for quick screening of V. parahaemolyticus in foods.     

High Salinity Relaying to Reduce Vibrio parahaemolyticus and Vibrio vulnificus in Chesapeake Bay Oysters (Crassostrea virginica)

Cases of Vibrio infections in the United States have tripled from 1996 to 2009 and these infections are most often associated with the consumption of seafood, particularly oysters (Crassostrea virginica). Information is needed on how to reduce numbers of Vibrio parahaemolyticus and Vibrio vulnificus in bi‐valve molluscan shellfish (for example, oysters). The purpose of this study was to evaluate the effectiveness of high salinity relaying or treatment in recirculating aquaculture systems (RASs) as methods to reduce the abundance of V. parahaemolyticus and V. vulnificus in oysters. For relaying field trials, oysters were collected from approved harvest waters, temperature abused outside under a tarp for 4 h, and then transferred to high (29 to 33 ppt.) and moderate (12 to 19 ppt.) salinities. For RAS treatment trial, oysters were transferred to 32 to 34 ppt. salinity at 15 °C. After 7, 14, 21, and in some instances 28 d, oysters were collected and analyzed for V. parahaemolyticus and V. vulnificus levels using multiplex real‐time PCR. Initial levels of V. parahaemolyticus and V. vulnificus ranged from 3.70 to 5.64 log₁₀ MPN/g, and were reduced by 2 to 5 logs after 21 to 28 d in high salinity water (29 to 34 ppt.). Oyster mortalities averaged 4% or less, and did not exceed 7%. Relaying of oysters to high salinity field sites or transfer to high salinity RAS tanks was more effective in reducing V. vulnificus compared with V. parahaemolyticus. These results suggest that high salinity relaying of oysters is more effective in reducing V. vulnificus than V. parahaemolyticus in the oyster species used in this study.     

Effect of X-ray treatments on inoculated Escherichia coli O157: H7, Salmonella enterica, Shigella flexneri and Vibrio parahaemolyticus in ready-to-eat shrimp

This study was conducted to evaluate the inactivation effect of X-ray treatments on Escherichia coli O157: H7, Salmonella enteric (S. enterica), Shigella flexneri (S. flexneri) and Vibrio parahaemolyticus (V. parahaemolyticus) artificially inoculated in ready-to-eat (RTE) shrimp. A mixed culture of three strains of each tested pathogen was used to inoculate RTE shrimp. The shrimp samples were inoculated individually with selected pathogenic bacteria then aseptically placed in sterile plastic cups and air-dried at 22 °C for 30 min (to allow bacterial attachment) in the biosafety cabinet prior to X-ray treatments. The inoculated shrimp samples were then placed in sterilized bags and treated with 0.1, 0.2, 0.3, 0.5, 0.75, 1.0, 2.0, 3.0 and 4.0 kGy X-ray at ambient temperature (22 °C and 60% relative humidity). Surviving bacterial populations were evaluated using a non-selective medium (TSA) with the appropriate selective medium overlay for each bacterium; CT-SMAC agar for E. coli O157: H7, XLD for S. enterica and S. flexneri and TCBS for V. parahaemolyticus. More than a 6 log CFU reduction of E. coli O157: H7, S. enterica, S. flexneri and V. parahaemolyticus was achieved with 2.0, 4.0, 3.0 and 3.0 kGy X-ray, respectively. Furthermore, treatment with 0.75 kGy X-ray significantly reduced the initial microflora on RTE shrimp samples from 3.8 ± 0.2 log CFU g⁻¹ to less than detectable limit (<1.0 log CFU g⁻¹).     

Effects of Lactic and Acetic Acid on Survival of Salmonella enteritidis During Refrigerated and Frozen Storage of Chicken Meats

Chicken leg and breast meat samples inoculated with Salmonella enteritidis [4–5 log most probable number (MPN)/cm²] were dipped into lactic acid (LA; 1% and 3%) and acetic acid (AA; 1% and 2%) solutions for 10 min. After packaging, samples were stored at 4 °C (10 days) or −18 °C (6 months). Immediately after dipping into 1% LA, 3% LA, 1% AA, and 2% AA solutions, S. enteritidis counts on leg meat samples were reduced by 0.75, 1.21, 0.85, and 0.95 log MPN/cm², while the reductions were 0.97, 1.72, 0.92, and 1.58 log MPN/cm² on breast meat samples, respectively. The differences between the water-washed control and the acid-treated groups for Salmonella counts were statistically significant (P < 0.05). Salmonella counts on leg meat samples treated with 1% LA, 1% AA, and 2% AA were reduced by 0.54–1.52 log MPN/cm² (P < 0.05) during storage at 4 °C. However, for the breast meat samples, only Salmonella counts of water-washed controls were significantly reduced during refrigerated storage (P < 0.05). S. enteritidis counts on organic acid-treated samples were reduced by 0.13–0.55 log MPN/cm² during storage at −18 °C for 6 months, while the reduction on the water-washed controls was 0.64 log MPN/cm². It can be concluded that lactic or acetic acid treatment could be useful especially for reducing the initial Salmonella contamination. On the other hand, this pathogen could survive on poultry meats during refrigerated and frozen storage even following lactic or acetic acid decontamination.     

Validation of high pressure processing for inactivating Vibrio parahaemolyticus in Pacific oysters (Crassostrea gigas)

This study identified and validated high hydrostatic pressure processing (HPP) for achieving greater than 3.52-log reductions of Vibrio parahaemolyticus in the Pacific oysters (Crassostrea gigas) and determined shelf life of processed oysters stored at 5 °C or in ice. Raw Pacific oysters were inoculated with a clinical strain of V. parahaemolyticus 10293 (O1:K56) to levels of 10^4-5 cells per gram and processed at 293 MPa (43 K PSI) for 90, 120, 150, 180 and 210 s. Populations of V. parahaemolyticus in oysters after processes were analyzed with the 5-tube most probable number (MPN) method. Negative results obtained by the MPN method were confirmed with a multiplex PCR detecting genes encoding thermolabile hemolysin (tl), thermostable direct hemolysin (tdh) and TDH-related hemolysin (trh). A HPP of 293 MPa for 120 s at groundwater temperature (8 ± 1 °C) was identified capable of achieving greater than 3.52-log reductions of V. parahaemolyticus in Pacific oysters. Oysters processed at 293 MPa for 120 s had a shelf life of 6-8 days when stored at 5 °C or 16-18 days when stored in ice. This HPP can be adopted by the shellfish industry as a post harvest process to eliminate V. parahaemolyticus in raw oysters.     

Effect of packaging conditions and temperature on viability of microencapsulated bifidobacteria during storage

In this study, Bifidobacterium longum B6 and B infantis CCRC 14633 were microencapsulated in various wall materials, including skim milk, gum arabic, gelatin and soluble starch. The stability of these microencapsulated bifidobacteria held at 25 or 4 °C in glass or polyester bottles with or without deoxidant and desiccant was determined. Microencapsulated cells of B longum B6 were generally more stable than the corresponding microencapsulated cells of B infantis CCRC 14633 under the various storage conditions tested. The presence of deoxidant and desiccant, especially at 25 °C, increased the survival of microencapsulated cells. Furthermore, the survival of bifidobacteria was enhanced when they were stored at 4 °C in glass bottles. It was also found that the wall material affected the survival of microencapsulated bifidobacteria. The viability of B longum B6 and B infantis CCRC 14633 was best when they were encapsulated in skim milk and held at 4 °C in glass bottles. Skim milk‐encapsulated B longum B6 cells showed a relatively low viability reduction of only 0.15–0.20 log (colony‐forming units (cfu g^?1)) after 42 days of storage at 4 °C in glass bottles, regardless of the presence of deoxidant and desiccant. A reduction of 0.38–0.76 log (cfu g^?1) was noted for skim milk‐encapsulated cells of B infantis CCRC 14633 under similar storage conditions. Copyright © 2004 Society of Chemical Industry     

Inhibition of selected pathogens inoculated on the surface of catfish fillets by high molecular weight chitosan coating

Antibacterial activity of high molecular weight water-soluble (HMWWS) chitosan (800 kDa) was investigated against four Gram-negative (Escherichia coli, Salmonella typhimurium, Vibrio cholerae and Vibrio parahaemolyticus) and two Gram-positive (Staphylococcus aureus and Listeria monocytogenes) bacteria. Catfish fillets were surface-inoculated with these food-borne pathogens and coated with chitosan dissolved in aspartic acid (AS) or acetic acid (AC) solution at different concentrations (1% or 3%). Samples were stored at 4 °C for 8 days, except for those inoculated with Vibrio species (10 °C for 6 days). Overall, the most effective coating treatment was the 3% HMWWS chitosan in AS solution (800AS3%). Compared with the control, significant (P < 0.05) reductions caused by 800AS3% were observed for all tested pathogens at the end of storage. The growth of V. parahaemolyticus was completely suppressed by 800AS3%. This study demonstrated that HMWWS chitosan in AS solution could be used as an alternative antimicrobial coating for catfish fillets.     

Rapid Detection of <Emphasis Type="Italic">Vibrio parahaemolyticus</Emphasis> in Shellfish by Real-Time Recombinase Polymerase Amplification

Vibrio parahaemolyticus (V. parahaemolyticus) is a zoonotic pathogen generally found in seafood. To detect the foodborne pathogen rapidly and accurately for food safety measures, we developed a real-time recombinase polymerase amplification (RPA) method. An evaluation of the specificity and sensitivity of the method is discussed here. A set of primers and probe was specially designed to target the tlh gene, which is usually regarded as a marker of total V. parahaemolyticus strains. During the reaction, target DNA was amplified and tagged with specific fluorophore within 10 min and at an incubation temperature of 40 °C. In addition to fast amplification and low temperature, the fluorescence signal was synchronized with the amplification of products for the generation of real-time data. The detection limit of this assay was 0.4 pg/μL of DNA, which is comparable to assays that use the bacterial culture as template, 4?×?10³ cfu/mL. The real-time RPA method had a stable performance when testing the spiking shellfish samples at the same level of contamination by the pathogen in different kinds of shellfish. Thus, the real-time RPA method shows great potential for on-site detection of V. parahaemolyticus, especially in low-resource settings.     

Spore-forming bacteria in commercial cooked,pasteurised and chilled vegetable purées

In commercial purées of broccoli, carrot, courgette, leek, potato and split pea, pasteurized in their final packaging and analysed at two periods, Bacillus spp. were the dominant aerobic mesophilic bacteria (AMB). Initial numbers were generally lower than 2 log cfu g⁻¹. They increased up to 6–8 log cfu g⁻¹after about 20 days of storage at 10°C. At 4°C, numbers of AMB after 20 days were lower than 3 log cfu g⁻¹in potato purée, lower than 4 log cfu g⁻¹in leek purée, and between 3 and 6 log cfu g⁻¹in other products. Strict anaerobes were in markedly lower numbers than AMB. At all storage temperatures tested courgette purée usually showed the most rapid bacterial growth and spoilage. On this product, an increase in storage temperature from 4°C to 10°C resulted in a threefold reduction in time to 5 log cfu g⁻¹, and time to spoilage. Growth kinetics of AMB in courgette purée at 20°C, 15°C, 10°C, 6·5°C and 4°C were determined using a mathematical model. Three hundred and forty eight isolates were identified using the API system. Bacillus circulans, B. macerans and B. polymyxa were among the main species isolated from products stored at 4°C and 10°C, while B. subtilis and B. licheniformis were the dominant species in product stored at abuse temperature. Bacillus cereus was isolated from all storage conditions, but mostly from products stored at abuse temperature.     

Prevalence and characterisation of Bacillus cereus in vacuum packed potato puree

Refrigerated processed foods of extended durability (REPFED) potato puree was analysed for Bacillus cereus contamination along the production line and during the product shelf‐life. Isolated B. cereus strains were tested for their psychrotrophic character and the ability to produce enterotoxins. Bacillus cereus contamination during four subsequent productions was in the range of 2.3–4.0 log cfu g^?1. Productions five and six were significantly less contaminated with B. cereus (≤1 log cfu g ^?1). All B. cereus isolates from the first four productions were able to grow at 7 ° and 10 °C, whereas the majority of the isolates from productions five and six did not. No B. cereus isolates grew at 4 °C. randomly amplified polymorphic DNA (RAPD) fingerprinting showed that the most of B. cereus contamination originated from one source. In total, 30.4% of isolates expressed enterotoxic character. The present study points out the necessity to prevent an ‘in house’ colonisation and contamination during food processing in order to accomplish the safety of REPFED throughout the shelf‐life. It also indicates the most critical steps in the production line of ready‐to‐eat potato puree and impact of failures regarding the food safety. The data provided can be used for risk assessment studies regarding B. cereus in REPFED.