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Salmonella enterica is a major cause of foodborne diseases, and is also an important pathogenic bacterium in poultry industry. Whole genome sequencing (WGS) has become a crucial molecular typing technology used for the surveillance of the pathogenic bacteria. In the present study, we adopted WGS for tracking transmission of S. enterica in the production chain of broiler chickens. A total of 74 S. enterica strains were isolated from the different steps of breeding and slaughtering in a large production enterprise in Sichuan Province, China. The isolation rate of Salmonella was the highest in procedure of defeathering (50.0%) and evisceration (36.7%). Serotype identification showed that 74 Salmonella isolates included 7 serotypes, among which Mbandaka accounted for the highest proportions (35.1%). WGS revealed that 74 strains belonged to 7 different sequence types (STs), as well as 7 different ribosomal STs and 35 core genome STs. cgMLST-based Minimum Spanning Trees and phylogenetic tree based on the SNPs indicated that three serotypes, Mbandaka, Indiana and Kentucky, could be clonally transmitted between broiler farm and slaughterhouse. Heterogeneous resistant phenotypes and genotypes were found in two serotypes, Indiana and Kentucky. Our study indicated WGS in an accurate tool for molecular typing of S. enterica. Routine surveillance of S. enterica in the production chain of broiler chickens is needed.

Current consumer preferences for both clean label food ingredients and convenience-based foods has provided a unique opportunity to explore the application of novel natural food preservatives in sous vide products. The anaerobic environment and relatively low thermal processing of the sous vide process creates a favorable environment for the survival, germination, and outgrowth of spore-forming bacterium Clostridium perfringens. The aim of this study was to identify effective novel natural ingredient formulations against C. perfringens and apply them within a vacuum-sealed sous vide chicken model exposed to abusive storage and chilling conditions. Among six commercial vinegar-based formulations, liquid vinegar with citrus extract (CE; 1.0%) and with lemon juice concentrate (LJC; 1.5%) were identified as the most effective at inhibiting three individual C. perfringens strains. Both reduced viable cell counts by 5 log CFU/mL (P 0.05), whereas reductions in spore counts ranged from 2 to 4 log CFU/mL depending on formulation and concentration used. Once incorporated to chicken meat 1.0% CE and 1.5% LJC before sous-vide cooking, completely inhibited the growth of mixed C. perfringens strains (P 0.05) during storage for 16 days at 12 and 16 °C. Exponential cooling from 54 to 4 °C was performed for 18 h to imitate abusive storage conditions. CE and LJC at 3.0% inhibited growth and reduced counts by 3.4 and 2.9 log CFU/g compared to respective controls. Treatments CE and LJC could be implemented within the formulation of a sous vide chicken product to provide an effective protection against C. perfringens meeting clean label expectations.

In many countries campylobacteriosis ranks as one of the most frequently reported foodborne illnesses and poultry is the commodity that is most often associated with these illnesses. Nevertheless, efforts to reduce the occurrence of pathogen contamination on poultry are often more focused on Salmonella. While some control measures are pathogen specific, such as pre-harvest vaccination for Salmonella, improvements in sanitary dressing and interventions applied during the slaughter process can be effective against all forms of microbial contamination. To investigate the potential effectiveness of these non-specific pathogen reduction strategies in the United States, it is helpful to assess if, and by how much, Campylobacter contamination of chicken meat has changed across time. This study assesses change considering data collected in both slaughter and retail establishments and comparing observed trends in contamination with trends in human surveillance data. The results support the assertion that substantial reductions in Campylobacter contamination of chicken meat in the late 1990s and early 2000s contributed to a reduction in the human case rate of campylobacteriosis. Further reductions in chicken meat contamination between 2013 and 2018 are more difficult to associate with trends in human illnesses, with one contributing factor being the inclusion of culture independent diagnostic test results in the official case counts during that time. Other contributing factors are discussed.

This study assessed changes in thermo-tolerance and capability to survive to simulated gastrointestinal conditions of Salmonella Enteritidis PT4 and Salmonella Typhimurium PT4 inoculated in chicken breast meat following exposure to stresses (cold, acid and osmotic) commonly imposed during food processing. The effects of the stress imposed by exposure to oregano (Origanum vulgare L.) essential oil (OVEO) on thermo-tolerance were also assessed. After exposure to cold stress (5°C for 5h) in chicken breast meat the test strains were sequentially exposed to the different stressing substances (lactic acid, NaCl or OVEO) at sub-lethal amounts, which were defined considering previously determined minimum inhibitory concentrations, and finally to thermal treatment (55°C for 30min). Resistant cells from distinct sequential treatments were exposed to simulated gastrointestinal conditions. The exposure to cold stress did not result in increased tolerance to acid stress (lactic acid: 5 and 2.5μL/g) for both strains. Cells of S. Typhimurium PT4 and S. Enteritidis PT4 previously exposed to acid stress showed higher (p

Biodegradable films reinforced with bio-nanomaterials are a solution for developing active packaging systems, shelf-life extension and protection of environment against conventional packaging. This study aimed to characterize the biocompatible chitosan (CS) films formulated with nano-liposomal garlic essential oil (NLGEO) and assess the physicho-mechanical, morphology properties and also microbial and chemical changes in chicken fillets during storage time at 4 °C. NLGEO was obtained by thin-layer hydration-sonication method using glycerol and tween 80 as plasticizer and emulsifier, respectively. Different levels (0, 0.5, 1 and 2%) of NLGEO with average size of ~101 nm were added into the chitosan matrix and films fabricated by casting method. The average size, polydispersity index and zeta potential were ~101 nm, 0.127 and −7.23, respectively. Control samples showed higher values for pH, total volatile nitrogen (TVN), peroxide value (PV), thiobarbituric acid-reactive substances (TBARS), and microbial count including total viable count (TVC), coliforms, Staphylococcus aureus and psychrotroph bacteria than treated samples. The films with higher NLGEO content represented stronger inhibitory effects. The incorporation of NLGEO improved the mechanical properties and water resistance of active films. Microstructure analysis also showed a nearly smooth surface morphology and homogenous structure with a good dispersion for NLGEO films. Significant synergistic effects in chemical and bacterial preservation of chicken fillet samples were observed by NLGEO films. The optimal mechanical and barrier properties of chitosan-NLGEO films introduced it a potential active packaging to extend the shelf life of chicken fillet.

In recent years, the on-farm prevalence of some poultry-related Salmonella serovars such as S. Kentucky, S. Heidelberg, S. Livingstone and S. Mbandaka has increased significantly, even replacing S. Enteritidis and S. Typhimurium as the most frequently isolated serovars in some production settings and countries. For this reason, the aim of this work was to determine the resistance to several stressing agents and food preservation technologies, in laboratory media and in egg products, of 4 strains of these emerging Salmonella serovars associated to poultry and poultry products and to make comparisons with 4 S. Enteritidis strains. First, the resistance to acid pH, hydrogen peroxide, NaCl, heat, HHP, PEF and UV of the 8 Salmonella strains studied was determined and compared in laboratory media. From this part of the study, it was concluded that variability in resistance to stress among the 8 studied strains varied depending on the investigated agent/technology. However, differences in resistance (2D-values) were always lower than 3.3-fold. Results obtained also indicated that the strains of the emerging serovars studied would display lower acid and NaCl resistance, higher heat resistance and similar oxidative, HHP, PEF and UV resistance than S. Enteritidis. Then, the resistance of these 8 strains was evaluated and compared in egg, egg products and poultry manure. For some agents -including osmotic stresses, UV and PEF- there was a very good correspondence between the results obtained in laboratory media and in real food matrices and poultry manure (r 0.85; p 0.01). A significant relationship was also found for acid and HHP resistance (p 0.05) and a trend for heat resistance (p 0.10). Therefore, in general terms, conclusions drawn from the study carried out in laboratory media - regarding intraspecific variability and the relative resistance of the different strains - might be extrapolated, although with caution, to real food scenarios. Results obtained in this investigation would help to better understand the physiology and ecology of Salmonella and to design better egg preservation strategies.

Methicillin-resistant Staphylococcus aureus (MRSA) is responsible for several difficult-to-treat infections and staphylococcal food poisoning (SFP). This study was conducted to investigate the prevalence and enterotoxigenicity of MRSA in broiler chicken meat and giblets. A total of 5.5% (8/144) of the examined samples were contaminated with mecA positive/mecC negative MRSA, with staphylococcal counts of approximately 102 colony forming units (CFU)/g in breast, leg and gizzard samples and approximately 3.3 × 103 CFU/g in frozen liver samples. Most MRSA isolates (75%, 6/8) harboured the staphylococcal enterotoxin B (seb) gene. Reverse transcription-PCR (RT-PCR) showed that MRSA isolates initiated SEB production in experimentally contaminated chicken livers within 24 h of storage at temperatures over 8 °C. SEB was maximally produced at 24 °C when the MRSA counts reached 7.3 × 103 ± 1.2 × 103 CFU/g sample homogenate. The current study concludes that the main broiler chicken MRSA isolates in Egypt harbour the seb gene. To mitigate possible SEB production, especially in broiler chicken livers, a maximum “out of refrigeration” time limit should be implemented for cold chain poultry products.

The prevalence of three pathogens in marinated chicken products and the evaluation of their quality by microbiological and sensory analysis were assessed. Eighty (80) samples obtained from several meat retail markets in Greece were analyzed for the presence of Campylobacter spp., Salmonella and Listeria monocytogenes. Concerning Campylobacter, rep-PCR and species specific PCR were applied for the differentiation and identification of isolates, respectively. The samples were subsequently stored aerobically at 4 °C for 5 days. Microbiological analysis, sensory assessment and HPLC analysis were carried out for the evaluation of spoilage microorganisms, sensory quality and the presence of preservatives (potassium sorbate and sodium benzoate). Τhe prevalence of Campylobacter spp., Salmonella, and Listeria monocytogenes was 50%, 11% and 44%, respectively. In the case of Campylobacter, from a total of 40 isolates, 27 were identified as Campylobacter coli, 4 as Campylobacter jejuni, whereas the remaining 9 belonged to unidentified Campylobacter species. Pseudomonas spp. was the dominant spoilage microbial genus in 43% of the samples, while in 31% of them a co-dominance of Pseudomonas spp. and Brochothrix thermosphacta was observed. Total aerobic counts increased to 7.0 log CFU/g at the 1st, 2nd or 3rd day of storage in 71% of the samples, while sensory analysis showed that 80% of the samples were characterized as spoiled after 3, 4 or 5 days. The presence of preservatives was confirmed in 31% of the samples and slightly affected the microbiological profile. In conclusion, the obtained data demonstrated the occurrence of foodborne pathogens and allowed the acquisition of an overall view about the microbiological quality of marinated chicken products.

Campylobacteriosis is a bacterial disease transmitted to humans through ingestion of contaminated food. Six hundred samples were collected, 200 from human stool samples, 200 from poultry products and 200 from poultry feces in Southern Brazil, and then inoculated on blood agar plates. A total of 58% of the poultry feces, 17% of the poultry meat, and 2% the of human stools tested positive for Campylobacter. Positive Campylobacter colonies were identified as Campylobacter jejuni or Campylobactercoli by multiplex PCR. Campylobacter isolates were analyzed using PFGE to compare different profiles according to the source. This study demonstrated that there are different Campylobacter clones distributed in different aviaries in Southern Brazil. In addition, PFGE molecular profiles suggested that broilers can be a source of contamination of poultry products. However, the human isolate studied did not show any relationship with other strains examined.

Danish organic broiler meat, represented by carcasses sampled at the end of processing after chilling, was more frequently contaminated with thermotolerant Campylobacter spp. than conventional broiler carcasses; the yearly mean prevalence being 54.2% (CI: 40.9-67.5) for organic and 19.7% (CI: 14.8-24.7) for conventional carcasses. Campylobacter jejuni was the most frequently isolated species. The difference in prevalence was obvious in all quarters of the year. Contamination of organic and conventional broiler carcasses was more likely to occur in the warmer summer months, in this case in the third quarter, as also documented for conventional broiler flocks. When contaminated, the mean concentration of Campylobacter on neck skin samples of organic and conventional carcasses was not significantly different (P=0.428); 2.0±0.65 log 10 cfu/g and 2.1±0.93 log 10 cfu/g, respectively. Assessing the relative risk of becoming ill following exposure to Campylobacter on conventional or organic broiler meat indicated that the risk per serving from organic carcasses was 1.7 times higher than that of conventional carcasses. The higher risk of illness from organic broiler carcasses compared with conventional broiler carcasses emphasizes the importance of implementing control measures in organic broiler production.

One of the most popular meat products of the local “cuisine” is sausage composed with 100% chicken or 100% pork. In this study, we aimed to determine the presence of Salmonella spp., Campylobacter spp. and Listeria spp. in chicken- and pork-sausages, quantify Salmonella spp. population and identify the factors that could be associated with contamination in the outlets. Two hundred and three batches of pork and chicken sausages were randomly collected from 67 local outlets (supermarkets, groceries and butcher shops). Salmonella spp. was detected in 11.8% (95% confidence interval (CI): [10.0; 13.5]) of samples, Campylobacter spp. in 1.5% [0.7; 4.2] and Listeria monocytogenes in 5.9% [4.4; 7.3]. Most probable number of Salmonella spp. varied between 6 cfu per gram to 320 cfu per gram. Salmonella serotypes isolated from pork and chicken sausages were S. Typhimurium (45.8%), S. London (20.8%), S. Derby (16.7%), S. Newport (8.33%), S. Blockley (4.2%) and S. Weltevreden (4.17%). Using a logistic (mixed-effect) regression model, we found that Salmonella spp. contamination was positively associated with sausages sold in papers or plastic bags and no control of rodents. Chicken sausages were associated with a decreasing risk of Salmonella contamination. Listeria monocytogenes contamination was positively associated with the presence of fresh rodent droppings in the outlet and negatively when the staff was cleaning regularly their hands with soap and water or water only. All the sampled outlets of Reunion Island were not equivalent in terms of food safety measures. Increasing awareness of these traders remains a cornerstone to limit the presence of Salmonella spp. and Listeria spp. in sausages, particularly in a tropical context (high temperature and humidity).

The objective of this study was to evaluate (eO)®, a biological time temperature integrator (TTI) as a quality and safety indicator for ground beef packed under modified atmosphere and spiced cooked chicken slices packed under modified atmosphere. Storage trials and challenge tests were thus performed on several batches of the studied food to monitor and model the behavior of Listeria monocytogenes, Salmonella, Staphylococcus aureus and the indigenous food flora. Then, two different prototypes of the TTI (eO)® were set and manufactured according to the studied products shelf lives. The TTI evolution with time at static and dynamic temperatures was monitored and modeled. Finally, exposure assessment models were set and used under several realistic storage profiles to assess the distributions of the concentration of the indigenous food flora and the distributions of the increase in the pathogens populations obtained at the end of the product shelf life or at the end point of the TTI, taking into account the TTIs batch variability. Results showed that in case of poor storage conditions, TTI can reduce the consumer exposure to altered or hazardous foods.

Epidemiological studies show that poultry meat and eggs are important sources for consumers' exposure to pathogens such as Salmonella and Campylobacter. There is a focus in many countries to reduce the level of human illness from food-borne pathogens. Reduction of the prevalence of contaminated poultry meat or eggs is one major area of focus. The other is risk communication to the consumer, where information aimed at changing the food preparation behaviour has been utilised as a risk management tool. The efficacy of messages such as 'cook poultry meat and eggs thoroughly' or 'wash your hands' will depend both on the ability to change consumer behaviour as well as where the risk can best be mitigated. In order to prioritise what message should be given to the consumer, the relative contribution of different exposure pathways finally leading to ingestion of the pathogens and resulting in illness needs to be known. It is important to know whether cross-contamination events or undercooking are the greatest risk lurking in consumers' kitchens. A review of studies looking at the location of pathogens in food products has been performed and data regarding internal and external (surface) contamination of poultry meat with Salmonella spp. and Campylobacter jejuni and C. coli is presented. In the case of eggs, data on internal contamination with Salmonella and for contamination of egg shells with Salmonella and Campylobacter are discussed. The results from published risk assessments for these pathogen-food commodity combinations have been evaluated and conclusions regarding the relative risk of internal and external contamination of poultry meat and eggs were drawn. In conclusion, cross-contamination events from activities such as use of the same cutting board for chicken meat and salad without intermediate cleaning or spreading of pathogens via the kitchen environment seem to be of greater importance than the risk associated with undercooking of poultry meat or eggs. Risk management options are discussed against the background of risk communication strategies used in different countries.

We describe the isolation and characterization of Shiga toxin (Stx)-producing Escherichia coli (STEC) O157:H7 from cooked and uncooked beef and chicken burgers and from chicken carcasses collected during sampling procedures in 2001 and 2002 in Buenos Aires City, Argentina. Of the 24 STEC O157:H7 strains isolated, 20 were recovered from 19 (6.8%) out of 279 samples of beef and chicken burgers, and 4 strains from 4 (10.3%) out of 39 chicken carcasses. The samples were analyzed following the USDA/FSIS 2002 method. The prevalent stx genotype was stx2 and stx2c (12 strains, 50%). All strains were characterized as eae and ehxA-positive. By XbaI-PFGE, the strains yielded 10 different patterns. Eighteen out of 24 strains were grouped in four clusters: #1 (4 strains, AREXHX01.0043), #2 (4 strains, AREXHX01.0022), #3 (8 strains, AREXHX01.0139), and #4 (2 strains, AREXHX01.0200). Identical strains by phage typing, stx genotyping and PFGE were detected in uncooked and cooked beef and chicken burgers in different restaurants, which had been collected on the same or different sampling dates. These findings help to underline the importance of STEC O157 detection in meat products, to improve active surveillance, and to define control strategies in order to prevent new cases of STEC infection.

Raw, frozen chicken nuggets/strips available at retail and prepared at home before consumption have been identified as a significant risk factor in contracting food-borne salmonellosis. Cases of salmonellosis from consumption of these products may be due, in part, to Salmonella strains originating in broiler feed. In this study the thermal resistances of Salmonella strains isolated from chicken nuggets and strips, chicken nugget/strip meat and broiler feed were determined to assess whether they exhibited enhanced thermal resistance. Thermal resistances (D- and z- values) of 7 cocktails (25 isolates, 4 serovars) were determined in commercially prepared irradiation-treated chicken nugget/strip meat blend, and heated in a constant temperature waterbath. The thermal resistances found were lower than those reported for similar strains in the literature. D-values ranged from 6.93 to 0.12 min at 55 and 62 degrees C respectively, with z-values from 4.10 to 5.17 degrees C. Two strains of S. Enteritidis separately isolated from pelleted feed and chicken nugget meat blend, with indistinguishable geno- and phenotypes, had lower (and probably identical) thermal resistances than the other isolates. Results indicated that the strains of Salmonella isolated from raw, frozen chicken nuggets/strips and pelleted broiler feed did not exhibit unusually high thermal resistance, and that normal heating (71 degrees C) prior to consumption should eliminate these organisms from chicken nuggets/strips.

Microbial risk assessment provides a means of estimating consumer risks associated with food products. The methods can also be applied at the plant level. In this study results of microbiological analyses were used to develop a robust single plant level risk assessment. Furthermore, the prevalence and numbers of Listeria monocytogenes in marinated broiler legs in Finland were estimated. These estimates were based on information on the prevalence, numbers and genotypes of L. monocytogenes in 186 marinated broiler legs from 41 retail stores. The products were from three main Finnish producers, which produce 90% of all marinated broiler legs sold in Finland. The prevalence and numbers of L. monocytogenes were estimated by Monte Carlo simulation using WinBUGS®, but the model is applicable to any software featuring standard probability distributions. The estimated mean annual number of L. monocytogenes-positive broiler legs sold in Finland was 7.2 × 106 with a 95% credible interval (CI) 6.7 × 106–7.7 × 106. That would be 34% ± 1% of the marinated broiler legs sold in Finland. The mean number of L. monocytogenes in marinated broiler legs estimated at the sell-by-date was 2 CFU/g, with a 95% CI of 0–14 CFU/g. Producer-specific L. monocytogenes strains were recovered from the products throughout the year, which emphasizes the importance of characterizing the isolates and identifying strains that may cause problems as part of risk assessment studies. As the levels of L. monocytogenes were low, the risk of acquiring listeriosis from these products proved to be insignificant. Consequently there was no need for a thorough national level risk assessment. However, an approach using worst-case and average point estimates was applied to produce an example of single producer level risk assessment based on limited data. This assessment also indicated that the risk from these products was low. The risk-based approach presented in this work can provide estimation of public health risk on which control measures at the plant level can be based.

Chicken meat is frequently contaminated with Campylobacter jejuni and is thought to be the major source of organisms causing human Campylobacter enteritis. Genotypic similarities between Campylobacter isolates from chicken meat at retail outlets and patients with gastroenteritis in Barbados suggested that it is a vehicle for infection of humans on the island and prompted this investigation of transmission of Campylobacter in a local poultry operation. Campylobacter testing was conducted at the hatchery, on the broiler farm and in the processing plant for two consecutive production cycles. The genetic relatedness of Campylobacter isolates was determined by RAPD typing with primer OPA 11. Hatchery samples and week-old chicks were negative for Campylobacter. Flocks became colonized as early as three weeks after introduction to the farm. Ten distinct RAPD genotypes were identified among isolates. Some genotypes were similar and may be of clonal origin. There was no evidence of vertical transmission of Campylobacter. The results suggest that the broiler flock was infected from more than one source in the farm environment.

Campylobacter causes bacterial diarrhoea in man and is a common foodborne pathogen, that has been associated mainly with poultry carcasses and processed poultry products as well as with drinking water. Genotyping of Campylobacter spp. from poultry was done in order to prove if predominant stable strains in the food chain are present. The influence of the slaughter process on the stability should be determined. Thermophilic Campylobacter spp. from eight poultry flocks were isolated from cloacal swabs, carcasses and offal at different abattoir processing steps to determine their stability. DNA-fingerprinting was done using Pulsed-Field Gel Electrophoresis (PFGE) with two enzymes (SmaI and KpnI) and ribotyping. More than 150 Campylobacter strains were ribotyped and these data were combined with the results of PFGE. Molecular typing showed that strains found in cloacal swabs before processing could also be isolated from carcasses and offal at different processing steps representing predominating stable strains. Strains with varying molecular pattern could additionally be detected at different processing steps. Both genotyping methods identified in agreement flock-specific strains. These remained stable through the slaughter of poultry and were not altered through the slaughter process. Despite the known genetic variability of thermophilic Campylobacter, stable predominant strains could be identified in the poultry slaughter process and those strains can thus enter the food chain.

It has been shown that the inappropriate use of antimicrobial household agents selects for organisms with resistance mechanisms (e.g. efflux pumps), which could lead to the development of antibiotic resistance. The reverse hypothesis, that antibiotic-resistant organisms become tolerant to other antibacterial agents (e.g. disinfectants) due to the action of efflux pumps, has however not been extensively examined. The objective of this study was to establish whether there is a link between antibiotic resistance in potential gastrointestinal pathogens and reduced sensitivity of these organisms to commonly used household antimicrobial agents. In this study, tetracycline and ofloxacin sensitive and resistant Escherichia coli (9 strains) and Salmonella spp. (8 strains) were isolated from poultry and clinical samples. In order to assess whether these bacteria had active efflux pumps, ethidium bromide accumulation assays were performed. Extrusion of the active components of three commercial household agents (triclosan, sodium salicylate, and ortho-phenylphenol) by efflux pumps was tested using spectrophotometric accumulation assays. In order to simulate the kitchen environment, in-use disinfectant testing using the commercial household agents was performed to determine changes in their efficacy due to antibiotic resistance. Active efflux pump activity and extrusion of all three active ingredients was observed only in the antibiotic resistant organisms. The antibiotic sensitive bacteria were also more susceptible than the resistant isolates to the household antimicrobial agents at concentrations below that recommended by the manufacturer. These resistant bacteria could potentially be selected for and result in hard to treat infections.

National and international health agencies have recommended a significant reduction in daily intake of sodium by reducing the amount of NaCl in foods, specifically processed meats. However, sodium reduction could increase the risk of survival and growth of spoilage and pathogenic microorganisms on these products. Therefore, alternate processing technologies to improve safety of sodium reduced foods are necessary. This study examined the effects of three different salt types and concentrations on high-pressure inactivation of Listeria monocytogenes in pre-blended ground chicken formulations. Ground chicken formulated with three salt types (NaCl, KCl, CaCl2), at three concentrations (0, 1.5, 2.5%) and inoculated with a four strain cocktail of L. monocytogenes (10(8) CFU g(-1)) were subjected to four pressure treatments (0, 100, 300, 600 MPa) and two durations (60, 180 s) in an experiment with factorial design. Surviving cells were enumerated by plating on Oxford agar and analysed by factorial ANOVA. Pressure treatments at 100 or 300 MPa did not significantly (P=0.19-050) reduce L. monocytogenes populations. Neither salt type nor concentration had a significant effect on L. monocytogenes populations at these pressure levels. At 600 MPa, salt types, concentrations and duration of pressure treatment all had a significant effect on L. monocytogenes populations. Formulations with increasing concentrations of NaCl or KCl showed significantly lower reduction in L. monocytogenes, while increase in CaCl2 concentration resulted in a significantly higher L. monocytogenes reduction. For instance, increase in NaCl concentration from 0 to 1.5 or 2.5% resulted in a log reduction of 6.16, 2.49 and 1.29, respectively, when exposed to 600 MPa for 60s. In the case of CaCl2, increase from 0 to 1.5 or 2.5% resulted in a log reduction of 6.16, 7.28 and 7.47, respectively. These results demonstrate that high-pressure processing is a viable process to improve microbial safety of sodium reduced poultry products.

Bacteriocins are amphiphilic peptides susceptible to adsorption to food macromolecules and proteolytic degradation. These properties may limit their use as preservation agents. The aim of the present work has been to elucidate the fate of the bacteriocin sakacin P in food. Nisin was used in a few experiments for comparison. Recovery of bacteriocins was studied in homogenates of cold-smoked salmon, chicken cold cuts and raw chicken, with verification of the results in the corresponding food products. More than 80% of the added sakacin P and nisin were quickly adsorbed to proteins in the food matrix. In foods that had not been heat-treated, proteolytic activity caused a rapid degradation of the bacteriocins, with less than 1% of the total activity left after 1 week in cold-smoked salmon, and even less in raw chicken. In heat-treated foods, the bacteriocin activity was stable for more than 4 weeks. The high fat content in salmon compared to chicken had no adverse effect on bacteriocin recovery or activity. However, mixing of triglyceride oils and bacteriocin solutions caused a considerable loss of activity. No principal differences were observed between sakacin P and nisin, but less nisin was adsorbed to muscle proteins at low pH, and the negative effect of oils was less pronounced for nisin. Growth of Listeria monocytogenes was completely inhibited for at least 3 weeks in both chicken cold cuts and cold-smoked salmon by addition of sakacin P (3.5 μg/g), despite the proteolytic degradation in the salmon.

The proportion of Campylobacter contaminated food and water samples collected by different surveillance systems often exhibit seasonal patterns. In addition, the incidence of foodborne campylobacteriosis also tends to exhibit strong seasonal patterns. Of the various product classes, the occurrence of Campylobacter contamination can be high on raw poultry products, and chicken is often thought to be one of the leading food vehicles for campylobacteriosis. Two different federal agencies in the United States collected samples of raw chicken products and tested them for the presence of Campylobacter. During the same time period, a consortium of federal and state agencies operated a nationwide surveillance system to monitor cases of campylobacteriosis in the United States. This study uses a common modeling approach to estimate trends and seasonal patterns in both the proportion of raw chicken product samples that test positive for Campylobacter and cases of campylobacteriosis. The results generally support the hypothesis of a weak seasonal increase in the proportion of Campylobacter positive chicken samples in the summer months, though the number of Campylobacter on test-positive samples is slightly lower during this time period. In contrast, campylobacteriosis cases exhibit a strong seasonal pattern that generally precedes increases in contaminated raw chicken. These results suggest that while contaminated chicken products may be responsible for a substantial number of campylobacteriosis cases, they are most likely not the primary driver of the seasonal pattern in human illness.

Campylobacter infection is one of the most common bacterial enteric pathogens. Campylobacter jejuni and Campylobacter coli infections are mostly food- and waterborne and especially poultry is often assumed to be an important source. The heat-stable serotyping system (the 'Penner' scheme) was used to study the serotype distribution of C. jejuni and C. coli isolated from different food products of poultry origin sampled from retail outlets in Denmark. A total of 156 isolates were serotyped, 85% of these were C. jejuni and 15% were C. coli. The most common C. jejuni serotypes were O:2 (30%), O:1,44 (12%) and the O:4-complex (8%). O:46 was the most frequent serotype among C. coli isolates. These serotypes are also common among Danish clinical isolates and isolates from broiler chickens and cattle. Differences in serotype distribution were seen for different kinds of poultry products. Isolates from chicken products covered a large selection of serotypes. In contrast, the majority of the isolates from other product groups (turkey, poussin, wild birds) were concentrated on 1-3 serotypes. Using the standard procedure for antigen preparation and serotyping, 25 of the 156 strains (16%) were nontypable. This rate of nontypable isolates is significantly higher than experienced for isolates from other sources than food products, i.e faecal samples from animals and humans. Subculturing and re-typing of the nontypable isolates improved the typability. After two, five and 10 subcultures 16, six and one isolate became typable, respectively. Only three isolates (2%) remained nontypable after 10 subcultures.

During four subsequent years (1993 until 1996), a study was conducted to isolate and characterize Salmonella in poultry carcasses and their products sold in Belgium. This was a semiquantitative approach (absence per l00 cm 2 or 25 cm 2 or 25 g and absence per cm 2 or g) to elucidate the degree of Salmonella contamination of the poultry. Serotyping was performed during the last two and a half years. Samples were frozen and kept at −20°C before analysis. This may have influenced the number of Salmonella recovered. No improvement in the rate of contamination was noted during these four years, with rates being 19.4% for 1993, 24.1% for 1994, 21.9% for 1995 and 36.7% for 1996. A 100% increase of Salmonella -positive samples resulted from cutting up the carcasses into individual parts. Chicken parts were more often contaminated with Salmonella than turkey parts. Boiling hen carcasses showed the highest Salmonella contamination. Prepared poultry, chicken parts and boiling hen carcasses are sometimes associated with Salmonella contamination levels of >1 cfu/cm 2 or g. In 1996, respectively 15.1%, 4.2% and 4.2% of highly contaminated samples (>1 cfu/cm 2 or g) were found for these product groups. The predominant three serotypes were S. enteritidis (16.3%), S. hadar (15.5%) and S. virchow (14.1%). S. newport was frequently isolated from turkey products.

The thermal inactivation of Salmonella enteritidis phage type 4 and Escherichia coli O157:H7 as affected by temperature (54.5-64.58C), pH (4.2-9.6 with HCl or NaOH) and NaCl concentration (0.5-8.5% w / w) was studied. Cell suspensions in modified tryptone soya broth were heated in a submerged-coil heating apparatus and survivors were enumerated on tryptone soya agar incubated aerobically. For most thermal inactivation data there was a logarithmic decrease in the viable cell concentration over the initial 4-6 log reduction and D-values were fitted. In some cases, tailing of the survivor curves was 10 observed with cells surviving longer than the D-values predicted. Models describing the effect of temperature, pH and NaCl concentration on the thermal inactivation of S. enteritidis and E. coli O157:H7 were produced. For both organisms, predicted z-values of 4.6-7.0 C8 were obtained depending on conditions, with larger z-values at higher levels of NaCl. Optimum survival occurred between pH 5 and pH 7 and increasing acidity or alkalinity caused a decrease in the predicted D-values. At equivalent pH, acetic acid and lactic acid (at 0.5, 1 and 2% w / w) generally had a similar, or increased, lethal effect compared with HCl, whereas in most cases citric acid had a less lethal effect. For E. coli O157:H7, increasing NaCl concentration had a protective effect up to the maximum tested (8.5% w / w), while for S. enteritidis optimal survival at a NaCl concentration of 5-7% w / w was predicted. The models were validated in foods by comparing predictions with published data. Most (80%) of the predicted D-values from the S. enteritidis model were within the 95% confidence interval (within 2.45-fold of the published data) for different Salmonella serotypes in whole egg, egg albumen, egg yolk, beef and milk. Most (93%) of the predicted D-values from the E. coli O157:H7 model were larger than the limited published data for this organism in meat, poultry, milk and apple juice with 42% within the 95% confidence interval (within 2.05-fold of the published data). The D-value models were incorporated into Version 1, and subsequent versions, of the predictive microbiology software program, Food MicroModel. © 1997 Elsevier Science B.V.

There is a need to prevent intestinal colonisation by Salmonella enteritidis and S. typhimurium in newly hatched chicks. Treatment with an undefined bacterial flora is not acceptable to regulatory agencies in some countries because of the potential risk of transmitting pathogens. A defined culture with a potency and stability equivalent to those of an undefined culture has not yet been developed. Since attenuated Salmonella vaccine strains could possess the colonisation characteristics but not the virulence of Salmonella wild-type strains, they could inhibit colonisation of the challenge organism. S. typhimurium live vaccines registered in Germany (Zoosaloral H, Salmonella vac®T), S. enteritidis aroA and S. typhimurium aroA strains, S. enteritidis, S. typhimurium and S. infantis wild-type strains or a competitive exclusion product (Broilact®) were used as pretreatment cultures and evaluated for their inhibitory effects against S. enteritidis and S. typhimurium colonisation in newly hatched SPF chickens. Day-old chicks were administered a pretreatment culture and infected orally with variants of S. enteritidis or S. typhimurium wild type-strains resistant to nalidixic acid or rifampicin 1 day after pretreatment. On days 2 and 6 after infection, viable numbers of the challenge strain in liver and caeca were determined. The results for birds pretreated with Broilact® showed a distinct protective effect against both S. enteritidis and S. typhimurium at a challenge dose of 104 cfu/bird. After pretreatment of chicks with S. enteritidis and S. typhimurium wild-type strains, the greatest degree of inhibition of caecal colonisation was produced using isogenic strains. Colonisation after infection with non-isogenic strains could not be prevented but only reduced for a brief period. These effects were also observed after administration of aroA strains of S. enteritidis and S. typhimurium but the protective effect was considerably lower than after pretreatment with wild-type Salmonella strains. Inoculation with attenuated S. typhimurium vaccines resulted in a weak but significantly reduced colonisation by S. typhimurium. Colonisation by S. enteritidis could not be diminished by either of the S. typhimurium vaccine strains. The results indicate in principle the potency of Salmonella vaccine strains to inhibit Salmonella wild-type colonisation in newly hatched chicks. Potential vaccine candidates should be tested for their capacity to prevent intestinal colonisation in newly hatched chicks.

The paper describes the use of the Delphi method to estimate the incidence of food-borne Salmonella in the UK and the effectiveness of alternative control measures. A panel of experts of food-borne Salmonella participated in the Delphi survey, which involved five rounds of questioning taking place in the period July 1993 to January 1994. Participants were asked to give initial estimates for a number of parameters and invited to revise these estimates through progressive rounds of the survey at which the group responses were reported back. This process resulted in a reduction in the variation between the estimates given by individual experts. The final estimated annual incidence of food-borne Salmonella in the UK was 537,000, although significant variation remained between, individual estimates. The foods judged to be the most important modes of transmission were poultry and poultry products (50% of cases) and eggs and egg products (26% of cases). The panel was also requested to estimate the effectiveness of strategies available to reduce the incidence of food-borne Salmonella from all sources. The most effective methods were judged to be food irradiation and mandatory application of HACCP, although there were significant differences in the judged effectiveness of these technologies for individual respondents. The paper demonstrates the efficacy of the Delphi method as a mechanism for reconciling differences between expert judgements of the incidence of food-borne disease and the effectiveness of alternative control strategies.

Clostridium perfringens growth from a spore inoculum was investigated in vacuum-packaged, cook-in-bag ground turkey (pH 6) that included 0.3% (w/w) sodium pyrophosphate, and sodium chloride at 0, 1, 2, or 3% (w/w). The packages were processed to an internal temperature of 71.1 degrees C, ice chilled and stored at various temperatures. The total C. perfringens population was determined by plating diluted samples on tryptose-sulfite-cycloserine agar followed by anaerobic incubation at 37 degrees C for 48 h. At 28 degrees C, the addition of 3% salt in turkey was effective in delaying growth for 12 h. At 15 degrees C, growth occurred at a relatively slow rate in the presence of 1-2% salt. Vegetative cells were not observed even after 28 days of storage in the presence of 3% salt. C. perfringens growth was not observed at 4 degrees C regardless of salt levels. The D-values ranged from 23.2 min (no salt) to 17.7 min (3% salt). Cyclic and static temperature abuse of refrigerated products for 8 h did not lead to growth by C. perfringens from a spore inoculum.

Risk profiles of pork and poultry meat were carried out using an Excel-based software program, Risk Ranger. It is a semi-quantitative risk estimator answering various questions relating to the probability of exposure to a hazard, susceptibility of the population of interest, severity of the illness caused by the hazard if present and probability of food containing an infectious dose. Therefore, qualitative and quantitative inputs were used to estimate and rank the risk of various hazards/food combinations. Risk scores provided by the tool were characterized as low, medium and high. Also, health risk was estimated separately, where needed, for low and high risk populations. Low risk scores were obtained for Salmonella spp., Listeria monocytogenes and enterohaemorrhagic Escherichia coli (EHEC) for low risk population. High risk scores were obtained for hepatitis E virus (HEV) in raw pork products (both low and high risk populations). Moderate risk scores for Salmonella spp. and L. monocytogenes in processed pork or poultry-meat products (ready-to-eat or to be reheated) and partially cooked pork products were also obtained (low risk population). Scores for Staphylococcus aureus, Clostridium perfringens and Bacillus cereus and various product types were mostly in the "medium" risk category, except for S. aureus/ready-to-eat pork products able to support growth of the organism, which fell into the high risk category. Campylobacter spp. gave moderate risk scores with one exception (raw poultry products), whereas Y. enterocolitica showed combinations of low risk and few of medium risk. High risk pathogen/product combinations identified were: 1) temperature abused, ready-to-eat pork and/or poultry-meat products with extended shelf life and cross-contaminated by L. monocytogenes (high risk population), EHEC (high risk population) or S. aureus (all population), 2) partially cooked or processed intended to be reheated pork products cross-contaminated by L. monocytogenes, served undercooked and receiving improper cooling or reheating (high risk population), and 3) all people consuming undercooked meals cross-contaminated with Campylobacter spp. (e.g. from raw poultry and raw poultry-meat products) and HEV (e.g. from raw pork and raw pork-meat products). Salmonellae gave high risk scores in all food categories (except preserved meat products) for high risk population. Preserved meats (mainly pork) such as dry fermented sausages gave low risk scores. Only Salmonella spp., L. monocytogenes and E. coli EHEC gave moderate risk ratings in case of ingredients likely to be contaminated at an early stage of processing (e.g. animal at slaughter) and inadequate fermentation process. These results may constitute a source of information for hazard assessment during application of a Food Safety Management System.

There have been significant outbreaks of listeriosis associated with consumption of contaminated ready-to-eat (RTE) turkey meat products. In this study, we investigated whether growth on RTE deli turkey meat sends environmental signals to listerial cells that makes them more virulent in the gastrointestinal tract of mice. L. Listeria monocytogenes strain F2365 grew from a starting inoculum of 10(3) CFU/mL to final numbers of 10(8)-10(9) CFU/mL (within 12 days at 10 degrees C) when inoculated onto sliced processed, or whole muscle, turkey breast, or into emulsified whole turkey breast. We did not observe any difference in the numbers of CFU recovered from the spleens and livers of A/J mice inoculated intragastrically with L. monocytogenes grown on sliced turkey meat, in emulsified turkey meat, or in brain heart infusion broth. These results suggest that growth on RTE sliced deli turkey, or in RTE emulsified deli turkey, does not enhance the ability of L. monocytogenes F2365 to cause gastrointestinal listeriosis in intragastrically challenged A/J mice.

We investigated the genotype diversity and dynamics of Campylobacter jejuni and Campylobacter coli in six commercial broiler farms during rearing and abattoir processing. In total, 223 C. jejuni and 36 C. coli strains isolated (on farm, transportation crates, carcasses after defeathering, and chicken wing meat at the end of the processing line) were subtyped by PCR-RFLP based on flagellin (fla typing) gene. Eleven (C. jejuni) and four (C. coli) different RFLP patterns were found. Multiple C. jejuni genotypes were identified in five out of six farms (except Farm 5). Furthermore, a clear tendency for dominance of particular genotypes was observed in almost all farms except Farm 3. Although diverse C. jejuni genotypes were isolated on the farms and transport crates, they were not detected in chicken wing cuts at the end of the processing line. We also observed varied distribution of types in different sampling stages both at the farm level and the processing environment. However, the interpretation of such fluctuations is precarious as new types occurred on some occasions, particularly during processing. Our results show that chicken wing meat contamination resulted mainly from farm strain carryover, and that the carcasses were probably contaminated during processing. In addition, the new strain types observed were isolated more frequently after defeathering as compared to other processing steps. Therefore, this stage, in addition to evisceration, is one of the critical control points in the processing line to prevent cross-contamination and for controlling the spread of campylobacters.

The isothermal survival curves of the heat resistant spores of Bacillus sporothermodurans IC4, in the range of 117–125 °C, were determined in chicken, mushroom and pea soups by the capillary method. They were all non-linear with a noticeable upper concavity and could be described by the equation log 10 [ N ( t )/ N 0 ]=− b ( T ) t n with a fixed power, n , of the order of 0.7–0.8. The temperature dependence of b ( T ) could be described by the equation b ( T )=log e {1+exp[ k ( T − T c )]}, where T c is the temperature where intensive inactivation starts and k is the slope of b ( T ) at temperatures well above T c . They were 121–123 °C and 0.2–0.4 °C −1 , respectively, depending on the soup. These parameters were used to estimate the survival curves of the spores in two non-isothermal heat treatments using the procedure originally proposed by Peleg and Penchina [Crit. Rev. Food Sci. Nutr. 40 (2000) 159]. The results were compared with experimental survival curves, determined by the direct injection method, in another laboratory. There was a general agreement, although not perfect, between the predicted and experimentally observed survival ratios. Also, the isothermal survival parameters, estimated directly from the non-isothermal inactivation data using the model, were in general agreement with those calculated from the isothermal data. This suggests that the heat inactivation patterns of B. sporothermodurans IC4 spores in soups can be at least roughly estimated using the same general survival model, which has until now only been experimentally validated for vegetative bacterial cells.

This study examined the incidence of Salmonella in Spanish poultry products. Samples included chicken carcasses, chicken parts (wings, legs and giblets—livers and hearts) and processed chicken products (red sausages, white sausages and hamburgers). The average detection rate was 49%, with the highest (55%) in chicken carcasses (skin) and the lowest (20%) in hamburgers. The chicken carcasses purchased in supermarkets were more contaminated (75%) than those from poulterers shops (25%). Salmonella Enteritidis, S. Poona, S. Paratyphi B and S. Worthington were isolated in 34.3%, 11.4%, 2.8% and 1.4% of the samples, respectively. One (1.4%) red sausage sample harboured two serotypes (S. Enteritidis and S. Worthington). This fact emphasizes the usefulness of subtyping several Salmonella isolates from the same sample in epidemiological studies.