A comparative study of sampling techniques for monitoring carcass contamination

Four bacteriological sampling techniques i.e. the excision, double swab, agar contract and modified agar contact techniques were compared by sampling pig carcasses before and after chilling. As well as assessing the advantages and disadvantages of the techniques particular attention was paid to variation due to the effects of chilling. The agar contact technique cannot be used for determining the contamination of carcasses because too many plates became overcrowded with micro-organisms. Results obtained by the double swab and modified agar technique gave standard deviations which were significantly higher than those of the excision technique. This may indicate a superior repeatability of the excision technique. Statistical analysis of the results showed that chilling influenced the differences between sampling techniques. The excision technique is to be regarded as the most suitable in view of its high accuracy and precision. When the technique is used properly, carcass damage is only small or negligible.     

Comparison of Three Sampling Techniques for Microbiological Analysis of Meat Surfaces

Efficiencies of three surface sampling techniques were compared for estimating total aerobic microflora, total coliform, fecal coliform and Escherichia coli. They were: stomaching excised skin (a destructive technique), direct agar contact and double moist swab (nondestructive techniques). Thirty ovine carcasses were evaluated for surface contamination, with sampling on three different dates. Difference between the three methods of sampling was significant for all bacterial groups evaluated. Stomaching excised skin recovered the highest number of bacteria. Moreover, significant differences in counts were observed with periods of sampling. For total aerobic microflora, counts appeared to be influenced by both sampling technique and sampling date.     

Comparison of sampling procedures for recovery of Listeria monocytogenes from stainless steel food contact surfaces

A number of techniques exist for microbiological sampling of food processing environments in food industries. In the present study the efficacies of nine sampling procedures for the recovery of Listeria monocytogenes from food contact surfaces, including a new sampling device consisting of a miniroller, were evaluated and compared. A stainless steel table was inoculated with L. monocytogenes strain 935 (serovar 4b, human origin) and L. monocytogenes strain 437/07 (serovar 1/2b, food origin), at 10(5) CFU/100 cm(2). L. monocytogenes strain 935 was best recovered with the minirollers (recovery of up to 6.27%), while poor recoveries (<0.30%) were obtained with the towel (one-ply composite tissue), alginate swab, metallic swab, and Petrifilm methods. In the case of L. monocytogenes strain 437/07 the replicate organism detection and counting (RODAC) ALOA contact plates yielded the best recoveries (4.15%), followed by the minirollers (up to 1.52%). Overall, recovery percentages with the minirollers were higher with stomacher homogenization than with Vibromatic agitation. The recovery percentages obtained for the Listeria strain of human origin were higher than those obtained with the food strain for all sampling procedures except Petrifilm and RODAC ALOA. With the miniroller device coated with wool fiber, the recovery of L. monocytogenes can be improved from 2 to 17 times over recoveries obtained with the sponge and cotton swab. This is the first report of a miniroller device for microbiological sampling in the available literature. The novel sampling procedure is convenient to apply on surfaces, is cost-effective, and results in better recovery of L. monocytogenes than do the conventional methods.     

Recovery and transfer of Salmonella typhimurium from four different domestic food contact surfaces

Domestic food contact surfaces can play an important role in the transmission of foodborne disease, yet debate continues as to which surface materials pose the greatest risk to consumer health in terms of cross-contamination during food preparation. Salmonella Typhimurium was inoculated onto stainless steel, Formica, polypropylene, or wooden surfaces (25 cm2) in the presence or absence of protein (tryptic soy broth supplemented with 5% horse serum) and held at room temperature. The pathogen was recovered from the test surfaces immediately after inoculation (T=0) and every hour for up to 6 h, by a conventional microbiological sampling technique and by direct transfer onto a model ready-to-eat food (cucumber slices). On all surfaces, pathogen numbers declined during the 6-h holding period, with the most rapid reductions occurring within the first hour. The presence of protein significantly increased (P < 0.05) the number of bacteria recovered from all surface types. However, regardless of application medium or holding time, the number of bacteria recovered from Formica (in all cases) and stainless steel (in most cases) was significantly higher than were the numbers on polypropylene or wood. Similarly, regardless of application medium or holding time, significantly higher bacterial numbers were transferred to the model food from Formica or stainless steel than from polypropylene or wooden surfaces. These differences were greater when the bacteria were applied in a protein-rich medium and the test surfaces held for 1 h or more. The results of this study emphasize that differences, both in recoverability and in the number of bacteria transferred to the model food rather than simply reflecting differences in pathogen survival, may also reflect differences in the ability of the test bacteria to remobilize from the different surface types. However, the results also demonstrate a fundamental problem when choosing food contact surfaces, i.e., that those characteristics that make a surface "easy to clean" may also render it more likely to release contaminating pathogens during common food preparation practices.     

A comparison of surface sampling methods for detecting coliforms on food contact surfaces

Newly developed self-contained swab-based methods were assessed for their ability to detect low levels of coliform bacteria on food contact surfaces. Their performance, under controlled laboratory conditions, was compared to a range of conventional microbiological methods, including traditional hygiene swabs, dipslides and sampling sponges. Stainless steel surfaces were inoculated with known levels of coliform bacteria and were sampled immediately after inoculation whilst still wet, or after they had been allowed to air-dry for 1 h. The sampling sponge was the least effective means of detecting coliforms on a wet surface, having a minimum detection limit of approximately 100 cfu cm⁻², whereas all other test methods were able to detect the presence of <3·5 cfu cm⁻². Allowing inoculated surfaces to dry resulted in a marked reduction in the minimum detection limits of all test methods although this was less for dipslides and the newly developed media-based hygiene swabs. A novel chemiluminescent swab-based assay was capable of providing results in 5 h, comparable to those obtained using traditional spread plates. The results of this study suggest that during the hygienic assessment of food contact surfaces when testing for coliforms, newly developed self-contained swab-based methods, including one giving results in 5 h, could prove a valuable tool for use within an integrated cleaning monitoring strategy.     

Swabbing the surface: critical factors in environmental monitoring and a path towards standardization and improvement

Abstract

Cross-contamination can be broadly defined as the transfer, direct or indirect, of microorganisms from a contaminated product to a non-contaminated product. Events that may result in cross-contamination include inadequate hygiene practices, contaminated equipment surfaces, contamination via food handling personnel, further product processing, or storage abuse All of these niches require consistent environmental surveillance systems to monitor microbial harborage sites to prevent foodborne illnesses via cross-contamination. Environmental surveillance is achieved through routine surface sampling of the food contact surfaces and surrounding areas. To better understand cross-contamination, the role of environmental surface transmission during outbreaks due to the presence and persistence of pathogenic microorganisms on various food contact surfaces must be investigated. However, studies on environmental sampling techniques are rarely performed in an actual food processing environment but rather under controlled variables within a laboratory-setting. Moreover, results and conclusions of studies differ because of the considerable variability across surface sampling tools due to individual operator dependency, low recovery rates, and low reproducibility. Information is also often lacking on environmental sampling tools used within a processing facility, the characterization of these tools, and the optimization of recovery of microorganisms for surface sampling. Thus, this review aims to: (1) discuss and compare factors impacting the recovery of microorganisms and the standardization of surface sampling methods for optimal recovery of microorganisms and (2) examine how research strategies could focus more towards the development of standard methodologies for surface sampling.     

Microbiological performance of a food safety management system in a food service operation

The microbiological performance of a food safety management system in a food service operation was measured using a microbiological assessment scheme as a vertical sampling plan throughout the production process, from raw materials to final product. The assessment scheme can give insight into the microbiological contamination and the variability of a production process and pinpoint bottlenecks in the food safety management system. Three production processes were evaluated: a high-risk sandwich production process (involving raw meat preparation), a medium-risk hot meal production process (starting from undercooked raw materials), and a low-risk hot meal production process (reheating in a bag). Microbial quality parameters, hygiene indicators, and relevant pathogens (Listeria monocytogenes, Salmonella, Bacillus cereus, and Escherichia coli O157) were in accordance with legal criteria and/or microbiological guidelines, suggesting that the food safety management system was effective. High levels of total aerobic bacteria (>3.9 log CFU/50 cm(2)) were noted occasionally on gloves of food handlers and on food contact surfaces, especially in high contamination areas (e.g., during handling of raw material, preparation room). Core control activities such as hand hygiene of personnel and cleaning and disinfection (especially in highly contaminated areas) were considered points of attention. The present sampling plan was used to produce an overall microbiological profile (snapshot) to validate the food safety management system in place.     

Characterizing bioaerosol risk from environmental sampling

In the aftermath of a release of microbiological agents, environmental sampling must be conducted to characterize the release sufficiently so that mathematical models can then be used to predict the subsequent dispersion and human health risks. Because both the dose-response and environmental transport of aerosolized microbiological agents are functions of the effective aerodynamic diameter of the particles, environmental assessments should consider not only the total amount of agents but also the size distributions of the aerosolized particles. However, typical surface sampling cannot readily distinguish among different size particles. This study evaluates different approaches to estimating risk from measurements of microorganisms deposited on surfaces after an aerosol release. For various combinations of sampling surfaces, size fractions, HVAC operating conditions, size distributions of release spores, uncertainties in surface measurements, and the accuracy of model predictions are tested in order to assess how much detail can realistically be identified from surface sampling results. The recommended modeling and sampling scheme is one choosing 3, 5, and 10 μm diameter particles as identification targets and taking samples from untracked floor, wall, and the HVAC filter. This scheme provides reasonably accurate, but somewhat conservative, estimates of risk across a range of different scenarios. Performance of the recommended sampling scheme is tested by using data from a large-scale field test as a case study. Sample sizes of 10-25 in each homogeneously mixed environmental compartment are sufficient to develop order of magnitude estimates of risk. Larger sample sizes have little benefit unless uncertainties in sample recoveries can be reduced.     

Transfer coefficient models for escherichia coli O157:H7 on contacts between beef tissue and high-density polyethylene surfaces

Risk studies have identified cross-contamination during beef fabrication as a knowledge gap, particularly as to how and at what levels Escherichia coli O157:H7 transfers among meat and cutting board (or equipment) surfaces. The objectives of this study were to determine and model transfer coefficients (TCs) between E. coli O157:H7 on beef tissue and high-density polyethylene (HDPE) cutting board surfaces. Four different transfer scenarios were evaluated: (i) HDPE board to agar, (ii) beef tissue to agar, (iii) HDPE board to beef tissue to agar, and (iv) beef tissue to HDPE board to agar. Also, the following factors were studied for each transfer scenario: two HDPE surface roughness levels (rough and smooth), two beef tissues (fat and fascia), and two conditions of the initial beef tissue inoculation with E. coli O157:H7 (wet and dry surfaces), for a total of 24 treatments. The TCs were calculated as a function of the plated inoculum and of the cells recovered from the first contact. When the treatments were compared, all of the variables evaluated interacted significantly in determining the TC. An overall TC-per-treatment model did not adequately represent the reduction of the cells on the original surface after each contact and the interaction of the factors studied. However, an exponential model was developed that explained the experimental data for all treatments and represented the recontamination of the surfaces with E. coli O157:H7. The parameters for the exponential model for cross-contamination with E. coli O157:H7 between beef tissue and HDPE surfaces were determined, allowing for the use of the resulting model in quantitative microbial risk assessment.     

The effect of nanometer dimension topographical features on the hygienic status of stainless steel

Wear of food contact surfaces through abrasion may increase the surface roughness and introduce different topographical features. Both of these properties may enhance retention of soil and microorganisms and affect the surface cleanability. To test this hypothesis, stainless steel surfaces with topographical features and surface roughness (Ra) values simulating those of worn in-use surfaces were prepared. Surfaces were imaged and Ra values determined using atomic force microscopy (AFM). These ranged from 23 to 900 nm. Surfaces were sprayed with standardized cell suspensions of Pseudomonas aeruginosa or Staphylococcus aureus and allowed to air dry and were then cleaned using a nonionic detergent delivered via a manual linear cleaning device. There was a 2-log reduction in numbers attached after cleaning, but there was no significant difference (P > 0.05) between the cleanability of the surfaces in terms of the numbers of cells per unit area remaining after cleaning, although cells appeared to be retained within topographical features. Thus, the simulated effect of wear of a hygienic food contact surface did not affect its cleanability after a one-off microbiological soiling event. AFM provided hitherto unavailable information on the topography of worn stainless steel surfaces. In future work, the surfaces will be repeatedly challenged with an organic soil-microorganism mixture after cleaning events, to provide a more rigorous, realistic test.     

Assessment of hygienic quality of surfaces in retail food service establishments based on microbial counts and real-time detection of ATP

Clean food contact surfaces are important in reducing the likelihood of foodborne disease transmission. The goal of this study was to assess and compare baseline cleanliness of food contact and environmental surfaces in retail food establishments by using ATP bioluminescence (ATP-B), visual assessment, and surface contact plates. Four hundred eighty-nine surface samples were collected from three food service establishments at the University of Minnesota-Twin Cities (Minneapolis) and analyzed for either ATP (252) or total aerobic plate count bacteria (237). ATP levels ranged from a minimum of 4 relative light units (RLU; 0.60 log RLU) on a clean slicer to a maximum of 506,618 RLU (5.77 log RLU) on a dirty cutting board. The overall mean was 1,950 RLU (3.29 log RLU). Cutting boards had the highest ATP levels (mean, 5,495 RLU or 3.74 log RLU; median, 6,761 RLU or 3.83 log RLU). Of the 128 samples judged visually clean at the time of sampling, 70.3 % failed ATP-B testing. Sixty-one (26 % ) of the 237 total aerobic plate count samples yielded counts of over 125 CFU/50 cm(2) (failed), and of those that failed, 40 % were assessed as visually clean before sampling. The highest average counts in CFU/50 cm(2) were found on slicers (104) and cutting boards (87). The results of this study suggest that the current practice of evaluating food contact surface cleanliness by sight and touch to meet regulatory requirements might be inadequate. ATP-B testing may be an efficient tool to facilitate creation, implementation, and validation of more effective food contact surface cleaning in food establishments.     

Identification of inadequately cleaned equipment used in a sheep carcass-breaking process.

Aeromonads deposited on pork during a carcass-breaking process were recovered on hydrophobic grid membrane filters placed on ampicillin-dextrin-ethanol agar. Isolates from 85 honey-yellow colonies from filters on that medium were Aeromonas hydrophila (95%) or Vibrio sp. (5%). Presumptive aeromonads, coliforms, and Escherichia coli in swab samples from product passing through a sheep carcass-breaking process were enumerated by hydrophobic grid membrane filtration techniques with a detection level of 1 CFU/100 cm2. Total aerobic counts were determined by a spread plate procedure with a detection level of 1 CFU/cm2 The numbers of aerobes, coliforms, and E. coli in the product were apparently unaffected by the carcass-breaking process, although coliforms and E. coli appeared to be redistributed from shoulder to loin and leg portions. However, the numbers of aeromonads on product increased by about two orders of magnitude as a result of the process. Few bacteria were recovered from most cleaned, large items of equipment, but aerobes, coliforms, and aeromonads were recovered at log total numbers of 5.25, 3.96, and 3.26, respectively, from most of 25 samples from bars supporting a conveyor belt. Also, aerobes, coliforms, E. coli, and aeromonads were recovered from 25 supposedly cleaned steel mesh gloves at log total numbers of 10.14, 5.54, 4.73, and 8.30, respectively. Those findings indicate that inspection of cleaned equipment and microbiological sampling of only food-contacting surfaces, as is the current practice at meat plants, cannot provide assurance that the cleaning of carcass-breaking equipment is adequate. Instead, enumeration of indicator organisms on product passing through a process seems to be required as well, with subsequent sampling of equipment to identify sources of contaminants if increases in numbers during processing are observed. For surety of adequate cleaning, enumeration of several types of indicator organism may be necessary, as increases during processing in the numbers of organisms that are present in relatively large numbers on product entering a process may be difficult to detect.     

国外禽畜胴体非破坏性微生物样品采集方法分析

屠宰生产线上禽畜胴体微生物样品采集方法分可破坏性采样和非破坏性采样。本文收集并分析了国际标准化组织(International Standardization Organization,ISO)、美国农业部、新西兰食品安全局以及澳大利亚检验检疫局关于禽畜胴体非破坏性微生物样品采集的方法,其包括湿-干拭子法(wet and dry swab method)、海绵拭子法(sponge sampling method)和纱布敷料法(gauze tampon method)、胴体冲洗采样法,不同屠宰工艺不同微生物目标其采样时机、采样位点、采样频率和微生物限量不尽相同,以期为我国禽畜产品加工生产和安全监控提供采样技术参考。     

Microbiological and chemical analyses of stainless steel and ceramics subjected to repeated soiling and cleaning treatments

Stainless steel and ceramic surfaces were subjected to repeated soiling and cleaning procedures, using a milk powder soil inoculated with Pseudomonas aeruginosa and Staphylococcus aureus, followed by spraying with water with or without 0.1% (vol/vol) nonionic detergent. Test surfaces were removed after 1, 5, 10, and 20 soiling-cleaning cycles and were analyzed for attached microorganisms (total viable count of bacteria removed by swabbing and percentage of coverage of cells plus soil stained with acridine orange). Surfaces were additionally examined using a range of analytical techniques: X-ray photoelectron spectroscopy (XPS), dynamic and imaging secondary ion mass spectroscopy (SIMS), and time-of-flight SIMS. Both microbiological and chemical analyses revealed an accumulation of material over the cycles to a maximum level. Surfaces were conditioned rapidly to saturation with organic material within one cycle (XPS), whereas fouling by microorganisms was less rapid (five cycles). Cleaning with detergent tended to retard the cumulative fouling process when compared with spray cleaning using water alone. The methods described provide a more realistic scenario for testing the cleanability of surfaces routinely found in food processing locations and for screening novel cleaning regimens and/or surface materials. The analytical techniques provide additional information on the kinetics of soiling, which complement the more familiar microbiological methods.     

Comparison of destructive and nondestructive sampling techniques of retail chicken carcasses for enumeration of hygiene indicator microorganisms

The type of sampling technique used to obtain food samples is fundamental to the success of microbiological analysis. Destructive and nondestructive techniques, such as tissue excision and rinsing, respectively, are widely employed in obtaining samples from chicken carcasses. In this study, four sampling techniques used for chicken carcasses were compared to evaluate their performances in the enumeration of hygiene indicator microorganisms. Sixty fresh chicken carcasses were sampled by rinsing, tissue excision, superficial swabbing, and skin excision. All samples were submitted for enumeration of mesophilic aerobes, Enterobacteriaceae, coliforms, and Escherichia coli. The results were compared to determine the statistical significance of differences and correlation (P < 0.05). Tissue excision provided the highest microbial counts compared with the other procedures, with significant differences obtained only for coliforms and E. coli (P < 0.05). Significant correlations (P < 0.05) were observed for all the sampling techniques evaluated for most of the hygiene indicators. Despite presenting a higher recovery ability, tissue excision did not present significant differences for microorganism enumeration compared with other nondestructive techniques, such as rinsing, indicating its adequacy for microbiological analysis of chicken carcasses.     

Impact of cleaning and disinfection on the non-culturable and culturable bacterial loads of food-contact surfaces at a beef processing plant

We assessed the impact of industrial cleaning and disinfection (C&D) on colony-forming units (CFUs), viable (culturable and viable but non-culturable) cells and on total cells (viable and dead cells). Bacterial loads on polyvinyl chloride (PVC) and stainless steel surfaces in a cutting room at a beef processing plant were determined before and after C&D by real-time PCR to quantify cells from successive swabs from surfaces with or without an ethidium monoazide pre-treatment and by CFU counts on tryptone soy agar. Agar contact plates were also applied after C&D for comparison. Before C&D, total cells reached 5.4 and 4.7logcells/cm(2), viable cells 4.0 and 4.4logcells/cm(2) and CFUs 3.1 and 2.9logCFU/cm(2) on PVC and stainless steel surfaces, respectively. Although C&D left surfaces visually clean, it did not lead to a significant reduction in total cells. Significant reductions were only observed on PVC for CFUs: 0.8 log and on stainless steel surfaces for viable cells and CFUs: 0.8 and 1.5 log, respectively. Our results show that CFUs were both more easily detached and killed on stainless steel surfaces than on PVC surfaces. Other important results include the following observations: 1) a single swabbing detached only between 2 and 27% of the actual bacterial load; 2) after C&D, the difference between the actual culturable population and the one assessed by one agar contact plate was 1.9 and 2.7logCFU/cm(2) on PVC and stainless steel surfaces, respectively.     

Attachment of enterohemorrhagic Escherichia coli to the surface of beef and a culture medium

This study was undertaken to evaluate the role of curli in the attachment of Enterohemorrhagic Escherichia coli (EHEC) to the surface of beef and a culture medium. Beef from an eye round roast and beef salami was sliced and cut into 3-mm thick, 4.6-cm diameter circles. The beef was exposed for different lengths of time to six EHEC cultures: the curli-expressing and noncurli-expressing variants of Escherichia coli 5-11 (O157:H7) and E. coli 7-52 (O103:H2) as well as a wild-type curli-expressing strain E. coli 7-57 (111:H-) and a noncurli-expressing strain E. coli 5-9 (O157:H7). The beef slices were rinsed following the exposure, and the EHEC cells that attached to the surfaces of the beef were enumerated. Two replications of each experiment were performed and the data generated was analysed statistically based on a 95% confidence level. In separate experiments, the EHEC cultures were grown on Luria-Bertani no salt agar at 28 °C for 72 h and the ease of removing the attached EHEC cells from the surface of the agar was evaluated. The results indicated that the curli-expressing cells had a firmer association with, and was more difficult to remove from, the surface of the agar in comparison to the noncurli-expressing cells. Cells of certain curli-expressing strains/variants of EHEC attached more efficiently under appropriate conditions, to the beef surface than did the cells of noncurli-expressing strains/variants. The differences in attachment by curliated and noncurliated cells were, however, minor. The study revealed the difficulties of studying EHEC attachment on biotic surfaces such as beef, suggesting that a better methodology is needed for more accurate evaluation of EHEC association with their contact surfaces.     

Cleanability of soiled stainless steel as studied by atomic force microscopy and time of flight secondary ion mass spectrometry

The hygienic status of food contact surfaces can deteriorate with wear. Effective cleaning regimes must remove any adsorbed organic material as well as microorganisms. Previous work has determined the extent of surface wear occurring on a stainless steel surface within the food industry, and we have reproduced representative samples in vitro. Two surface analytical techniques, atomic force microscopy and time of flight secondary ion mass spectrometry were combined with fluorescence microscopy to give detailed analysis of stainless steel surfaces fouled with starch and milk powder, then cleaned with water either by a spray or brushing method. It was found that the surface cleanability is affected by the cleaning regime and the surface roughness, not only the average vertical roughness but also by the shape of the surface defects, with sharp scratches more difficult to clean than wider surface defects. Spray cleaning with distilled water was found to be a selective method by preferentially removing proteinaceous material more easily than fatty acid ester material. The analytical techniques employed provided information on selective cleanability and surface topography at a hitherto unexplored level, and the information gained may be of value in the design and investigation of novel cleaning regimes and hygienic surfaces.     

A microbiological survey of fresh meat in the supermarket trade. Part 1: Carcasses and contact surfaces

Sanitary conditions were monitored at eleven supermarkets (from two major chains, Sup groups A & B) in the South African butcher industry. The variables measured were the temperature of carcasses and environments, the muscle pH of carcasses and the microbiological status of carcasses and human or personal and equipment contact surfaces (chillers-three surfaces, delivery trucks-five surfaces, cutting rooms-16 surfaces). All supermarkets, except one, received beef carcasses, distributed by one of two wholesale organizations from the same abattoir. No consistency was found in the contamination level of different parts of carcasses at different supermarkets, although there was a tendency for fore quarters to be more contaminated than hind quarters. Chiller surfaces, delivery truck surfaces and equipment had significantly different microbial counts at the different supermarkets. Sup group B received carcasses with higher mean surface temperatures and microbial counts than those of the carcasses received by Sup group A, but the mean microbial count of equipment was lower than that encountered at Sup group A. The result was less contaminated meat at the supermarkets in Sup group B, illustrating how a combination of the microbial quality of carcasses received by such supermarkets, and the sanitary programme in operation at such supermarkets affects the contamination level of retail premises. It could furthermore be shown that personnel surfaces (hands and clothes) and equipment like saws and mincers are consistent contributors to contamination at the retail level of the meat industry.