Quantification of Campylobacter Species Cross-Contamination during Handling of Contaminated Fresh Chicken Parts in Kitchens

Numerous outbreak investigations and case-control studies for campylobacteriosis have provided evidence that handling Campylobacter-contaminated chicken products is a risk factor for infection and illness. There is currently extremely limited quantitative data on the levels of Campylobacter cross-contamination in the kitchen, hindering risk assessments for the pathogen commodity combination of Campylobacter and chicken meat. An exposure assessment needs to quantify the transfer of the bacteria from chicken to hands and the kitchen environment and from there onto ready-to-eat foods. We simulated some typical situations in kitchens and quantified the Campylobacter transfer from naturally contaminated chicken parts most commonly used in Germany. One scenario simulated the seasoning of five chicken legs and the reuse of the same plate for cooked meat. In another, five chicken breast filets were cut into small slices on a wooden board where, without intermediate cleaning, a cucumber was sliced. We also investigated the transfer of the pathogen from chicken via hands to a bread roll. The numbers of Campylobacter present on the surfaces of the chicken parts, hands, utensils, and ready-to-eat foods were detected by using Preston enrichment and colony counting after surface plating on Karmali agar. The mean transfer rates from legs and filets to hands were 2.9 and 3.8%. The transfer from legs to the plate (0.3%) was significantly smaller (P < 0.01) than the percentage transferred from filets to the cutting board and knife (1.1%). Average transfer rates from hands or kitchen utensils to ready-to-eat foods ranged from 2.9 to 27.5%.     

Cross-contamination in the kitchen: estimation of transfer rates for cutting boards, hands and knives

Aims: To quantify cross-contamination in the home from chicken to ready-to-eat salad. Methods and Results: Based on laboratory scenarios performed by de Jong et al. (2008) , transfer rates were estimated for Campylobacter jejuni and Lactobacillus casei as a tracer organism. This study showed that transfer characteristics for both micro-organisms were comparable when washing regimes and transfer via items (cutting board, hands and knives) were compared. Furthermore, the study showed that the use of separate transfer rates for transfer from chicken to items and from items to salad will lead to an overestimation of campylobacteriosis risk. Applying good hygienic practices resulted in final levels of bacteria in the salad below the detection limit. Our study showed that it is important to include these data points in model fitting. Conclusions: Results obtained in observational studies with Lact. casei can be translated to Camp. jejuni using the transfer rates obtained in this study. Cross-contamination by hands, cutting boards and knives was equally important. Significance and Impact of the Study: Cross-contamination should be incorporated in microbiological risk assessments. The present study contributes to this by quantifying transfer of Camp. jejuni and Lact. casei from raw chicken via various contact surfaces into the ready-to-eat product.     

Enumeration of Campylobacter spp. on the surface and within chicken breast fillets

AIM: To investigate how many Campylobacter bacteria are present on the surface and inside chicken breast fillets, with a focus on generating data distributions which can be used in risk assessments for this pathogen-commodity combination. METHODS AND RESULTS: We analysed 100 fresh retail chicken breast fillets (skinless and deboned) by means of a rinse sample for surface and 55 fillets for internal pathogen contamination using 10 g meat and a most probable number technique. Prevalence was 87% on the surface and 20% in the deep tissue. The mean number of Campylobacter on the surface of the fillets was 1903 CFU, with a median of 537 CFU and a maximum of 38,905 CFU. Campylobacter counts inside the tissue were <1 CFU g(-1) meat (mean = 0.24 CFU, median = 0.15 CFU, maximum = 0.74 CFU). In addition, we investigated the influence of the type of package on the occurrence of the pathogen. Data provide an indication of less favourable conditions for survival of the pathogen on chicken meat packed under a modified atmosphere of carbon dioxide in nitrogen, in comparison with ambient air or vacuumed packages. CONCLUSIONS: Given the high numbers of the pathogen on the chicken meat surface in comparison with low levels of internal contamination, it can be concluded that cross-contamination during the preparation of contaminated chicken is a more important pathway for consumers' exposure to Campylobacter than the consumption of undercooked meat. Significance AND IMPACT OF THE STUDY: The detailed quantitative data on the occurrence of C. jejuni and C. coli on the surface and inside chicken meat presented here can be useful for future probabilistic exposure assessments.     

Transfer of Campylobacter jejuni from raw to cooked chicken via wood and plastic cutting boards

Aims: We quantified Campylobacter jejuni transferred from naturally contaminated raw chicken fillets and skins to similar cooked chicken parts via standard rubberwood (RW) and polyethylene cutting boards (PE). Methods and Results: RW and PE cutting boards (2·5 × 2·5 cm²) were constructed. RW surfaces were smooth and even, whereas PE was uneven. Scoring with scalpel blades produced crevices on RW and flaked patches on the PE boards. Raw chicken breast fillets or skin pieces (10 g) naturally contaminated with Camp. jejuni were used to contaminate the cutting boards (6·25 cm²). These were then briefly covered with pieces of cooked chicken. Campylobacter jejuni on raw chicken, the boards, and cooked chicken pieces were counted using a combined most‐probable‐number (MPN)‐PCR method. The type of cutting board (RW, PE; unscored and scored) and temperature of cooked chicken fillets and skins were examined. Unscored PE and RW boards were not significantly different in regards to the mean transfer of Camp. jejuni from raw samples to the boards. The mean transfer of Camp. jejuni from scored RW was significantly higher than from scored PE. When the chicken fillets were held at room temperature, the mean transfer of Camp. jejuni from scored RW and PE was found to be 44·9 and 40·3%, respectively. Conclusions: RW and PE cutting boards are potential vehicles for Camp. jejuni to contaminate cooked chicken. Although cooked chicken maintained at high temperatures reduced cross‐contamination via contaminated boards, a risk was still present. Significance and Impact of the Study: Contamination of cooked chicken by Camp. jejuni from raw chicken via a cutting board is influenced by features of the board (material, changes caused by scoring) and chicken (types of chicken parts and temperature of the cooked chicken).     

Prevalence of Campylobacter spp., Escherichia coli, and Salmonella serovars in retail chicken, turkey, pork, and beef from the Greater Washington, D.C., area.

A total of 825 samples of retail raw meats (chicken, turkey, pork, and beef) were examined for the presence of Escherichia coli and Salmonella serovars, and 719 of these samples were also tested for Campylobacter spp. The samples were randomly obtained from 59 stores of four supermarket chains during 107 sampling visits in the Greater Washington, D.C., area from June 1999 to July 2000. The majority (70.7%) of chicken samples (n = 184) were contaminated with Campylobacter, and a large percentage of the stores visited (91%) had Campylobacter-contaminated chickens. Approximately 14% of the 172 turkey samples yielded Campylobacter, whereas fewer pork (1.7%) and beef (0.5%) samples were positive for this pathogen. A total of 722 Campylobacter isolates were obtained from 159 meat samples; 53.6% of these isolates were Campylobacter jejuni, 41.3% were Campylobacter coli, and 5.1% were other species. Of the 212 chicken samples, 82 (38.7%) yielded E. coli, while 19.0% of the beef samples, 16.3% of the pork samples, and 11.9% of the turkey samples were positive for E. coli. However, only 25 (3.0%) of the retail meat samples tested were positive for Salmonella. Significant differences in the bacterial contamination rates were observed for the four supermarket chains. This study revealed that retail raw meats are often contaminated with food-borne pathogens; however, there are marked differences in the prevalence of such pathogens in different meats. Raw retail meats are potential vehicles for transmitting food-borne diseases, and our findings stress the need for increased implementation of hazard analysis of critical control point (HACCP) and consumer food safety education efforts.     

Antimicrobial-resistant Campylobacter species from retail raw meats

The antimicrobial susceptibilities of 378 Campylobacter isolates were determined. Resistance to tetracycline was the most common (82%), followed by resistance to doxycycline (77%), erythromycin (54%), nalidixic acid (41%), and ciprofloxacin (35%). Campylobacter coli displayed significantly higher rates of resistance to ciprofloxacin and erythromycin than Campylobacter jejuni, and Campylobacter isolates from turkey meat showed a greater resistance than those from chicken meat.     

Microbiological examination of ready-to-eat cold sliced meats and pâté from catering and retail premises in the UK

AIMS: To establish the microbiological quality of cold ready-to-eat sliced meats and paté from catering and retail premises, and investigate links hypothesized between foodborne Campylobacter infection and the consumption of cold sliced meats. METHODS AND RESULTS: A total of 4078 cold meat and paté samples were collected and examined according to a standardized protocol. Comparison with published microbiological guidelines revealed that most ready-to-eat meat and paté samples (75%) were of satisfactory/acceptable microbiological quality and 25% were of unsatisfactory/unacceptable quality. Two cold meat samples (<1%) were of unacceptable microbiological quality because of the presence of Campylobacter jejuni in 25 g and Listeria monocytogenes at 3.4 x 104 CFU g-1. CONCLUSIONS: Acceptable microbiological quality was associated with premises where the management was trained in food hygiene and those that had hazard analysis in place. Poor microbiological quality was associated with storage above 8 degrees C, presliced meats, infrequent cleaning of slicing equipment and poor control of practices that may lead to cross contamination. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides important information about the microbiological quality of cold ready-to-eat meats and paté. It also assists caterers, retailers, enforcement officers and policy makers to understand how different food safety practices affect microbiological quality.     

Recovery of Campylobacter jejuni and Campylobacter coli from inoculated foods by selective enrichment.

A direct enrichment procedure was developed to selectively recover small numbers of Campylobacter jejuni, C. coli, and nalidixic acid-resistant thermophilic Campylobacter from foods. The procedure includes an enrichment medium composed of brucella broth, 7% lysed horse blood, 0.3% sodium succinate, 0.01% cysteine hydrochloride, vancomycin (15 micrograms/ml), trimethoprim (5 micrograms/ml), polymyxin B (20 IU/ml), and cycloheximide (50 micrograms/ml) that is inoculated with 10 or 25 g of food and incubated with agitation under microaerophilic conditions at 42 degrees C for 16 to 18 h. After incubation, the medium is plated directly onto Campy-BAP agar plates (M. J. Blaser et al., Ann. Intern. Med. 91:179-185, 1979), and resulting colonies that resemble Campylobacter are identified by conventional tests. The foods evaluated included raw milk, hamburger, and chicken skin which had aerobic plate counts of 10(5) to 10(9) bacteria/g. The procedure was effective in recovering as few as 0.1 cell of Campylobacter per g of food. Of the 50 isolates of Campylobacter evaluated, all were recovered from raw milk and hamburger at a level of 1 to 4 cells/g, and 41 and 40 isolaes were recovered from the hamburger and milk, respectively, at 0.1 to 0.4 cell/g. The enrichment was least effective for recovering campylobacters from chicken skin, as 7 and 26 of 50 isolates were not recovered at 1 to 4 and 0.1 to 0.4 cell/g, respectively. This new procedure is more rapid, direct, and effective than other enrichment or direct plating procedures for recovering small numbers of campylobacters from foods.     

Campylobacter contamination in French chicken production from farm to consumers. Use of a PCR assay for detection and identification of Campylobacter jejuni and Camp. coli

AIMS: Campylobacter contamination in French chicken production from the farm to the consumer was determined using a PCR assay for bacteria detection and identification. METHODS AND RESULTS: Samples were bird droppings from poultry houses, neck skins, livers, hearts, gizzards, wings, legs and escalopes from slaughterhouses and gizzards, legs, drumstick, breast and escalopes from a supermarket. Bacterial DNA extraction was performed after an enrichment step in a broth and was followed by PCR. An internal control (IC) was used for both DNA extraction and PCR. Campylobacter were detected in 79.2% of poultry houses. Of the 303 samples, 201 were Campylobacter-positive (i.e. 66.3%) including 43.2% faecal samples, 5.6% slaughterhouse samples and 17.5% supermarket samples. There was no significant difference between the molecular method and the conventional culture technique for Campylobacter detection whatever the samples. The sensitivity was 5 UFC g(-1) of samples and 1.5 x 10(3) UFC ml(-1) of enrichment broth. The use of IC revealed PCR inhibition in 13 samples and problems in the DNA extraction in five samples. CONCLUSION: Significant Campylobacter contamination affects all stages of French chicken production. SIGNIFICANCE AND IMPACT OF THE STUDY: The understanding of Campylobacter contamination at different levels of chicken production and the determination of the best place(s) for intervention are important for significantly decreasing Campylobacteriosis. Our technique is rapid and can be used on different chicken samples for Campylobacter detection and identification.     

Cross-contamination in the kitchen: effect of hygiene measures

Aims:  To determine the effect of hygiene measures on cross-contamination of Campylobacter jejuni at home and to select a safe tracer organism for C. jejuni.
Methods and Results:  Comparative tests were conducted with nonpathogenic Escherichia coli and Lactobacillus casei and L. casei was chosen as the safe tracer organism. Salads containing chicken breast fillet contaminated with a known number of C. jejuni and L. casei were prepared according to different cross-contamination scenarios and contamination levels of salads were determined. Cross-contamination could be strongly reduced when cleaning cutting board and cutlery with hot water (68°C), but generally was not prevented using consumer-style cleaning methods for hands and cutting board.
Conclusions:  Dish-washing does not sufficiently prevent cross-contamination, thus different cutting boards for raw meat and other ingredients should be used and meat–hand contact should be avoided or hands should be thoroughly cleaned with soap. Lactobacillus casei can be used as a safe tracer organism for C. jejuni in consumer observational studies.
Significance and Impact of the Study:  Cross-contamination plays an important role in the transmission of food-borne illness, especially for C. jejuni . This study delivers suitable data to quantitatively assess the risk of campylobacteriosis caused by cross-contamination and it shows the effect of different preventive hygiene measures.     

Microbiological examination of ready-to-eat stuffing from retail premises in the north-east of England. The 'Get Stuffed' survey

AIMS: To establish the microbiological quality of ready-to-eat stuffing from retail premises in the north-east of England. To establish threshold levels of bacteria in the product for acceptance as a ready-to-eat food. To determine the relationship between the microbiology of the product and production processes. METHODS AND RESULTS: A microbiological study of ready-to-eat stuffing using validated methods was performed on 147 samples from 139 retail premises. The determinants investigated were as follows: aerobic colony count, Escherichia coli, Enterobacteriaceae, Staphylococcus aureus, Bacillus spp., Salmonella spp. and Campylobacter spp. Results indicate that using current guidelines 76.3% were satisfactory, 15.6% were acceptable and 8.2% were of unsatisfactory quality. CONCLUSIONS: Unsatisfactory results were due to high aerobic colony counts, E. coli, Enterobacteriaceae and S. aureus. There were significant associations between bacteriological quality and temperature of storage, food hygiene training, product discard policy and confidence in management scores. SIGNIFICANCE AND IMPACT OF THE STUDY: The microbiology of ready-to-eat stuffing suggests that this is a relatively safe product. It is suggested that the product be placed in food category 3 in the current guidelines for ready-to-eat foods.     

Comparison of Genotypes and Antibiotic Resistances of Campylobacter jejuni and Campylobacter coli on Chicken Retail Meat and at Slaughter

Multilocus sequence typing (MLST) and antibiotic resistance patterns of Campylobacter jejuni and Campylobacter coli from retail chicken meat showed high overlap with isolates collected at slaughterhouses, indicating little selection along the production chain. They also showed significant common sequence types with human clinical isolates, revealing chicken meat as a likely source for human infection.     

Antimicrobial-Resistant Campylobacter Species from Retail Raw Meats

The antimicrobial susceptibilities of 378 Campylobacter isolates were determined. Resistance to tetracycline was the most common (82%), followed by resistance to doxycycline (77%), erythromycin (54%), nalidixic acid (41%), and ciprofloxacin (35%). Campylobacter coli displayed significantly higher rates of resistance to ciprofloxacin and erythromycin than Campylobacter jejuni, and Campylobacter isolates from turkey meat showed a greater resistance than those from chicken meat.     

Survival of Salmonella on refrigerated chicken carcasses and subsequent transfer to cutting board

Objective:  To determine the effect of refrigeration time and temperature on Salmonella cell numbers on inoculated chicken carcasses and their transfer to a plastic cutting board.
Methods and Results:  The survival of Salmonella on chicken skin and the transfer to a plastic cutting board when exposed to different refrigeration temperatures (2, 6 or 8°C) for 9 days were the two main issues on which this work focused. Two scenarios were carried out to ascertain these effects: carcasses treated with a decontaminating acetic acid solution and untreated carcasses. All of the contaminated carcasses remained contaminated after 9 days of refrigeration. However, on untreated samples, while Salmonella numbers increased almost 1·5 log at 8°C, the pathogen numbers decreased about 1 log at 2 and 6°C. On acid-treated samples, cell numbers slightly decreased at all of the temperatures studied. Temperature did not affect salmonellae transfer to the cutting board, but time did. Acid decontamination increased cell numbers transferred to the cutting board compared with untreated samples.
Conclusion:  Proper refrigeration at low temperatures did not allow Salmonella numbers to rise, regardless of which carcasses had been, or had not been, acid treated. Despite the fact that the rate of transfer was not affected by temperature, the acid treatment detached Salmonella cells from the chicken skin and, therefore, the probability of greater cross-contamination should be studied further.
Significance and Impact of the Study:  The results of this study may provide better information about the refrigeration conditions for fresh chicken storage and also determine if these, along with acetic acid decontamination of broiler chicken, would affect the pathogen transfer to a cutting board.     

Specific discrimination of chicken DNA from other poultry DNA in processed foods using the polymerase chain reaction

In the present study, specific discrimination of chicken DNA contamination in processed foods using the polymerase chain reaction was investigated. The primer pair was designed to amplify a 102-bp fragment of the chicken mitochondrial 16S ribosomal RNA gene. While the DNA from chicken meat was amplified, the DNA from other poultry meat, mammalian meat, fish, shellfish, and cereals was not amplified. The primer amplified DNA fragments derived from model processed and nonprocessed food samples containing 0.001, 0.01, 0.1, 1, 10, and 100% chicken.     

Specific Discrimination of Chicken DNA from Other Poultry DNA in Processed Foods Using the Polymerase Chain Reaction

In the present study, specific discrimination of chicken DNA contamination in processed foods using the polymerase chain reaction was investigated. The primer pair was designed to amplify a 102-bp fragment of the chicken mitochondrial 16S ribosomal RNA gene. While the DNA from chicken meat was amplified, the DNA from other poultry meat, mammalian meat, fish, shellfish, and cereals was not amplified. The primer amplified DNA fragments derived from model processed and nonprocessed food samples containing 0.001, 0.01, 0.1, 1, 10, and 100% chicken.     

Killing of Campylobacter on contaminated plastic and wooden cutting boards by glycerol monocaprate (monocaprin)

Aims: Contamination in the kitchen with foodborne bacteria is a risk factor in human exposure to these pathogens, an important route being transfer of bacteria from contaminated cutting boards and other surfaces to humans. The aim of this study was to test microbicidal emulsions of glycerol monocaprate (monocaprin) against Campylobacter on contaminated cutting boards. Methods and Results: Plastic and wooden cutting boards, soiled with meat juice heavily contaminated with Campylobacter, were treated for 2 min with emulsions of monocaprin (MC) made in water or in buffer at low pH. Viable Campylobacter counts were reduced below the detectable level on plastic board surfaces after treatment with MC emulsions with or without 1·25% washing‐up liquids (WUL). The counts were also greatly reduced on wooden boards (P < 0·05). Conclusions: Monocaprin emulsions and mixtures of MC emulsions and WUL may be useful as sanitizers/disinfectants in kitchens and in other food preparing and processing facilities. Significance and Impact of the Study: Cleaning with MC emulsions with or without WUL may reduce the risk of human exposure to Campylobacter.     

Enteropathogenic bacteria in frozen chicken.

Eighty-two samples of frozen chicken from retail stores were examined for the presence of Campylobacter, Yersinia enterocolitica, and salmonellae. Aerobic plate counts and numbers of coliform bacteria at 37 degrees C were determined. Campylobacter fetus subsp. jejuni was found in 22% of the samples, Y. enterocolitica was found in 24.5% and Salmonella typhimurium was found in one sample (1.2%). The isolated strains of Y. enterocolitica belonged to serotypes 4, 5b, 6, and 8. Aerobic plate counts and numbers of coliform bacteria at 37 degrees C were not found to be noticeably higher in samples containing pathogens than in pathogen-free samples. This investigation showed that chicken does contain other pathogenic bacteria than salmonellae. Campylobacter and Y. enterocolitica were isolated in much higher frequencies than Salmonella.     

The microbiological quality of washing-up water and the environment in domestic and commercial kitchens

AIMS: To determine the microbiological quality of washing-up water and the environment in domestic and commercial kitchens. METHODS AND RESULTS: Chicken meals were prepared by people without food safety training in their own kitchen (n = 52) or by trained staff in a commercial kitchen (n = 10). Study participants then washed-up, cleaned the kitchen and completed a food hygiene questionnaire. The temperature and microbiological quality of the washing-up water, and the presence of pathogens in dishcloths, tea towels and other kitchen samples was determined. Of the raw chickens used in meal preparation, 96 and 13% were naturally contaminated with Campylobacter or Salmonella spp., respectively. In domestic kitchens, two of 45 sponges, dishcloths or scourers and one of 32 hand- or tea towels were contaminated with Campylobacter after washing-up and cleaning but none of the tap or sink swabs yielded pathogens. The mean washing-up water temperature in the domestic kitchens was 40.7 degrees C, whereas in the commercial kitchen it was 44.7 degrees C (P = 0.04). Study participants who used hotter water (>/=40 degrees C) had lower levels of bacteria in their washing-up water. The aerobic plate counts of the washing-up water samples in domestic homes were usually between 105 and 106 CFU ml-1 but those associated with the commercial kitchen were consistently lower (P = 0.01). Despite this, Campylobacter was detected in one of 10 washing-up water samples from the commercial kitchen but in none of the samples from domestic kitchens. CONCLUSIONS: Pathogenic microorganisms can be recovered relatively frequently from the kitchen environment. SIGNIFICANCE AND IMPACT OF STUDY: By identifying factors that affect the number of microorganisms in washing-up water and the kitchen environment, evidence-based recommendations on implementing domestic food hygiene can be made.