Comparison of swab transport media for recovery of Listeria monocytogenes from environmental samples

Environmental monitoring is recognized as an important strategy for controlling Listeria monocytogenes in food processing facilities. Samples are taken by swabbing environmental surfaces, and the swabs are immersed in a medium for transport to the laboratory. In this study, buffered peptone water (BPW), Dey-Engley neutralizing broth (DE), neutralizing buffer (NB), Letheen broth (LE), and newly described MCC buffer (MCC) were evaluated as transport media for recovery of sanitizer-stressed L. monocytogenes from inoculated swabs. After storage at 4°C, the media performed similarly, but at 25°C relative recovery efficiency from the inoculated sponges was DE > LE > BPW > MCC > NB. Recoveries from stainless steel surfaces followed similar trends. MCC, DE, and NB were compared for L. monocytogenes recovery in the presence of Escherichia coli, Enterococcus faecalis, Lactobacillus plantarum, Pseudomonas fluorescens, and Listeria innocua. After 4°C storage, all population levels changed little; after 25°C storage, DE allowed the best growth of L. monocytogenes regardless of other species present. MCC, DE, and NB performed similarly for recovery of L. monocytogenes from an artificial milk biofilm and for recovery of Listeria spp. from swabs obtained from a meat processing facility. Transport medium formulation, time and temperature of swab storage, and coexistence of other species affect recovery of sanitizer-stressed L. monocytogenes from environmental swabs. The study confirms the need to maintain 4°C storage conditions during swab transport.     

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.     

Comparison of surface sampling methods and cleanability assessment of stainless steel surfaces subjected or not to shot peening

The cleanability of AISI 304 stainless steel surfaces, indicated by the removal of Escherichia coli cells or Aspergillus niger spores was assessed by controlled inoculation and washing treatment of samples in standardised conditions. Two systems of recapture (Rodac plate technique and swabbing technique) were compared. Four industrial finishes, subjected or not to shot peening, contaminated at low concentration (1–10 cfu/cm²), and then washed with distilled water or alkaline detergent, were examined. The Rodac plate technique detected most of microorganisms inoculated (80% for E. coli cells and 67% for A. niger spores), whereas the swabbing technique recovered only 1% of the E. coli cells and 26% of the A. niger spores. Using the Rodac plate technique E. coli cells proved to be easily detachable from samples either with distilled water (98%) or alkaline detergent (>99%). For the surfaces contaminated with A. niger spores, the cleanability increased from 34% with distilled water to 77% with alkaline detergent. In these working conditions type of finish (shot treated or not) had no significant effect on cleanability of stainless steel.     

Surface material, temperature, and soil effects on the survival of selected foodborne pathogens in the presence of condensate

The effects of surface type (stainless steel, acetal resin, and fiberglass reinforced plastic wall paneling [FRP]), soil, and temperature on the survival of Listeria monocytogenes, Salmonella spp., and Yersinia enterocolitica, in the presence of condensate were evaluated. Surface coupons--half soiled with sterile porcine serum--were exposed to cell suspensions made from individual five-strain cocktails composed of organisms from the same genus (10(7) CFU/ml) in Butterfield's phosphate buffer and incubated for 2 h at 25 degrees C allowing attachment of cells to coupon surfaces. Coupons were rinsed to remove unattached cells, incubated at either 4 or 10 degrees C under condensate-forming conditions, and sampled at six time intervals over a 15-day period. For enumeration, cells were removed from the coupons by vigorous shaking in 100 ml of Butterfield's phosphate buffer with 3 g of glass beads and plated on tryptic soy agar with 0.6% yeast extract. Stainless steel did not support the survival of Listeria as well as acetal resin or FRP. Acetal resin and stainless steel were less supportive of Salmonella than FRP. All surfaces supported the survival of Yersinia over the 15-day trial equally. Temperature had little effect on survival of all organisms across all surfaces with one exception. However, Yersinia displayed growth on FRP at 10 degrees C. but death at 4 degrees C. Serum had a protective effect on L. monocytogenes on all surfaces, with populations sustained at significantly (P < or = 0.05) higher numbers over time than unsoiled coupons. Serum didnot effect survival of Salmonella or Yersinia on stainless steel, acetal resin, or FRP.     

Use of one-ply composite tissues in an automated optical assay for recovery of Listeria from food contact surfaces and poultry-processing environments

A novel one-ply composite tissue (CT) method using the Soleris (formerly BioSys) optical analysis system was compared with the conventional U.S. Department of Agriculture (USDA) environmental sponge enrichment method for recovery of Listeria from food contact surfaces and poultry-processing environments. Stainless steel and high-density polyethylene plates were inoculated to contain a six-strain L. monocytogenes cocktail at 10(4), 10(2), and 10 CFU per plate, whereas samples from naturally contaminated surfaces and floor drains from a poultry-processing facility were collected with CTs and environmental sponges. CT samples were transferred into Soleris system vials, and presumptive-positive samples were further confirmed. Sponge samples were processed for Listeria using the USDA culture method. L. monocytogenes recovery rates from inoculated stainless steel and polyethylene surfaces were then compared for the two methods in terms of sensitivity, specificity, and positive and negative predictive values. No significant differences (P > 0.05) were found between the two methods for recovery of L. monocytogenes from any of the inoculated stainless steel and polyethylene surfaces or environmental samples. Sensitivity, specificity, and overall accuracy of the CT-Soleris for recovery of Listeria from environmental samples were 83, 97, and 95%, respectively. Listeria was detected 2 to 3 days sooner with the CT-Soleris method than with the USDA culture method, thus supporting the increased efficacy of this new protocol for environmental sampling.     

Comparison of 3M Petrifilm environmental Listeria plates against standard enrichment methods for the detection of Listeria monocytogenes of epidemiological significance from environmental surfaces

ABSTRACT:  Environmental monitoring using sensitive methods for detection and elimination of harborage sites of Listeria monocytogenes is key to the control of this organism. The 3M™ Petrifilm™ Environmental Listeria (EL) Plate—a no enrichment method—was compared with the USDA/FSIS, modified USDA/FSIS (mUSDA), and ISO methods for detection/recovery of L. monocytogenes on 4 environmental surfaces (brick, dairy board, stainless steel, and epoxy resin). The efficacy of 3 sampling devices including the Microbial-Vac system^®, environmental sponge, and 3M Quick swab in recovering epidemiologically significant strains of uninjured and sublethally injured L. monocytogenes from environmental surfaces was evaluated. Environmental surfaces were inoculated with Listeria to obtain final cell densities of approximately 10 to 100 CFU/100 cm². The surfaces were then sampled and processed. For all methods, percent recovery (% samples where Listeria was detected) was significantly higher ( P < 0.05) for uninjured cells (75% to 100%) compared to injured cells (58.9% to 81.1%). The Petrifilm EL Plate method efficiently recovered both low level and injured Listeria populations from environmental test surfaces when used in conjunction with environmental sponge and the 3M Quick swab sampling. The mUSDA method was superior to all other methods for recovering both uninjured (100% recovery) and injured L. monocytogenes (80.8% to 81.1% recovery). Sponges and swabs were equally effective in recovering uninjured and injured Listeria and were significantly different ( P < 0.05) from the Microbial-Vac system. The findings indicate that both mUSDA and Petrifilm EL Plate methods can be used for the detection of potentially injured Listeria on food processing environmental surfaces.     

Influence of peroxyacetic acid and nisin and coculture with Enterococcus faecium on Listeria monocytogenes biofilm formation

Biofilm formation is a matter of concern in food industries because biofilms facilitate the survival of pathogenic bacteria such as Listeria monocytogenes, which may contaminate food-processing equipment and products. In this study, nisin and two Enterococcus faecium strains were evaluated for their effect on biofilm formation by L. monocytogenes cultured in brain heart infusion broth and on stainless steel coupons. Elimination of preformed L. monocytogenes biofilms by peroxyacetic acid also was tested. Adhesion control experiments were performed with pure cultures of L. monocytogenes after swab collection of adhered cells, which were then enumerated on PALCAM agar plates and visualized by scanning electron microscopy. Formation of a biofilm was recorded when the number of adhered cells was at least 10(3) CFU/cm2. When L. monocytogenes was cocultured with E. faecium bac-, the number of adhered L. monocytogenes cells was 2.5 log lower (P = 0.002) when initially compared with the control culture, but after 6 h of incubation a biofilm was again detected. However, in coculture on stainless steel coupons, E. faecium bac+ inhibited L. monocytogenes adherence and did not allow biofilm formation for up to 48 h (P < 0.001). In the presence of nisin or after treatment with peroxyacetic acid, bacterial growth was reduced (P < 0.001) up to 4.6 and 5.6 log CFU/cm2, respectively, when compared with L. monocytogenes cultures on untreated coupons. However, after these treatments, cells were still present, and after 24 h of incubation, a renewed biofilm was detected in L. monocytogenes cultures treated with nisin. Although all tested conditions reduced L. monocytogenes growth to some extent, only coculture with E. faecium bac+ efficiently reduced biofilm formation, suggesting a potential control strategy for this pathogen.     

Formation of biofilms by Listeria monocytogenes under various growth conditions

Eight strains of Listeria monocytogenes (7644, 19112, 15313, Scott A, LCDC, 10403S, SLCC, and 1370) produce biofilms when grown on polyvinyl chloride microtiter well plates. The growth medium (tryptic soy broth [TSB] or modified Welshimer's broth [MWB] at 32 degrees C) influenced the amount of biofilm formed; maximum biofilms were formed in MWB by six strains and in TSB by the remaining two strains. This result suggests that the growth medium is critical in development of L. monocytogenes biofilm. This organism also produced biofilms on stainless steel chips. Biofilm formation on these chips was observed following growth in TSB at 4, 20, and 37 degrees C. After 20 h of incubation at 20 or 37 degrees C, the cell density was approximately 10(6) CFU per chip, and after 4 days incubation at 4 degrees C, the cell density was 10(5) CFU per chip. L. monocytogenes strain Scott A biofilm formation on stainless steel chips was visualized using scanning electron microscopy, which revealed dense aggregates of cells held together by meshlike webbing.     

Use of enterocin CCM 4231 to control Listeria monocytogenes in experimentally contaminated dry fermented Hornád salami

The effectiveness of enterocin CCM 4231 in controlling Listeria monocytogenes contamination in dry fermented Hornád salami was examined. Three independent salami treatments were conducted under pilot plant and laboratory conditions. Salamis were produced according to standard technological parameters and stages with ripening for 3 weeks. The reference samples consisted of the meat mixture without either L. monocytogenes or bacteriocin addition. The control sample (CS) consisted of the meat mixture with 1% of L. monocytogenes inoculum (10(8) cfu ml(-1)) added; while the experimental sample (ES) consisted of the same mixture with enterocin CCM 4231 (12800 AU g(-1)) added. Sampling was done on the first day of the experiment, before and after bacteriocin addition for ES, on the second day and after 1, 2 and 3 weeks. The enterocin addition resulted in the reduction of L. monocytogenes by 1.67 log cycle in the ES when compared to the CS immediately after addition of the bacteriocin. Although on the second day, the growth of L. monocytogenes in ES reached 3.38 cfu g(-1) (log 10), a difference of 1.72 log was found between the ES and the CS. After 1 week of ripening, the L. monocytogenes count in the CS reached 10(7) cfu g(-1); while in the ES the count was 10(4) cfu g(-1), a difference which was maintained after 2 and 3 weeks of ripening. However, bacteriocin activity in the ES could not be detected analytically. The meat mixture used did not contain Listeria.     

Survival of Listeria monocytogenes Scott A on metal surfaces: implications for cross-contamination

Listeria monocytogenes is an important re-emerging pathogen which is commonly found in the environment. Many outbreaks have been associated with the contamination of food produce, often linked to cross-contamination from surfaces or equipment to prepared foodstuffs. In the present study a number of copper-base metal alloys have been used to assess the survival times of L. monocytogenes on different materials, in comparison with stainless steel. High concentrations (10(7)) of bacteria were placed on metal coupons cut from each alloy. After defined incubation times, coupons were placed in tubes containing phosphate buffered saline and vortexed to remove the cells. Aliquots were then plated onto tryptone blood agar plates and the number of colony forming units counted. The high concentration of bacteria was used to represent a "worst-case" scenario. The results indicate that survival is greatly reduced on a copper-base alloy compared to stainless steel. Viable cells could be detected on stainless steel after 24 h incubation at room temperature. On copper, brass, aluminium bronze and silicon bronze, no viable bacteria could be detected after 60 min incubation, indicating a 5 log reduction (the detection limit of the procedure was 100 bacteria). No cells could be detected from copper nickel and copper nickel zinc alloys, after 90 min incubation. The viability stain, 5-cyano-2,3-ditolyl tetrazolium chloride (CTC), confirmed these results, with actively respiring bacteria being clearly labelled on stainless steel after 24 h. The results suggest that careful choice of surface material could reduce the potential risk of cross-contamination in industrial, commercial and domestic environments.     

An evaluation of sampling methods for the detection of Escherichia coli and Salmonella on Turkey carcasses

The efficacy of rinse, excision, and swab methods for the microbiological analysis of prechill turkey carcasses was investigated. Aerobic plate counts from a 50-cm2 area of the breast sampled by excision and by swabbing were compared. Escherichia coli and Salmonella recoveries were determined from turkeys sampled by a carcass rinse (CR), a modified rinse with the carcass supported in a swing (MCR), a two-site swab of 50 cm2 at the back and thigh (2S), a one-site swab of 50 cm2 beneath the wing (1S), a whole-carcass swab of the inner and outer carcass surface (WS), and excision of 25 g of neck skin tissue (NE). The effect of diluent volume (25, 50, and 100 ml) on E. coli counts from swab samples was also assessed. The aerobic plate count from breast tissue sampled by excision was greater than that by swabbing (P < 0.05). E. coli recoveries by the MCR method were similar to those by CR. E. coli counts from IS and WS samples were higher when swabs were stomached in 50 rather than 25 ml of diluent (P < 0.05). For swabs stomached in 50 ml of diluent, E. coli recoveries by the MCR, 2S, 1S, and WS methods were similar. For swabs stomached in 50 ml of diluent, Salmonella recoveries by the WS and MCR methods were higher than those by the 2S and 1S methods. Excision was more effective than swabbing for obtaining total bacterial counts from reduced turkey carcass areas. Whole-carcass sampling by rinsing or swabbing is necessary for optimum Salmonella recovery. Sampling a reduced area of the carcass is sufficient for E. coli analysis.     

Strains and suspending menstrua as factors affecting death and injury of Listeria monocytogenes during freezing and frozen storage

Cell suspensions of Listeria monocytogenes strains V7, California, and Ohio in phosphate buffer solution, tryptose broth, or milk were frozen and stored at -18 degrees C. At appropriate intervals during storage, a sample was thawed at 35 degrees C and surface-plated on suitable media to allow colony formation by noninjured or noninjured plus injured cells. Degrees of death and injury were calculated from the data. Cells of L. monocytogenes were more resistant to death and injury when they were suspended in milk or tryptose broth rather than phosphate buffer solution. There was a significant (two-way ANOVA) difference in resistance to death and injury during frozen storage among strains of L. monocytogenes suspended in tryptose broth. The difference was nonsignificant when the cells were suspended in phosphate buffer solution or milk. Listeria monocytogenes strain Ohio was more resistant to death and injury during frozen storage when cells were suspended in tryptose broth rather than milk. The opposite was true for strains V7 and California. Death and injury of L. monocytogenes strains V7, California, and Ohio suspended in phosphate buffer solution were 98.7, 97.9, and 91.2% and 77.5, 51.6, and 70.2%, respectively, after 4 wk of frozen storage. The values were 67.3, 91.6, and 42.3% and 44.4, 65.6, and 32.6%, respectively, when cells were suspended in tryptose broth, and they were 37.8, 40, and 60.7% and 10.8, 66.8, and 46%, respectively, when cells were suspended in milk.     

Water activity of bacterial suspension media unable to account for the baroprotective effect of solute concentration on the inactivation of Listeria monocytogenes by high hydrostatic pressure

Inactivation of Listeria monocytogenes (10(8) CFU/ml) by high hydrostatic pressure (HHP) from 400 to 600 MPa at 25 degrees C for 10 min was investigated with various concentrations of sodium chloride, sucrose, and sodium phosphate buffer solutions. Sodium chloride significantly inhibited HHP-induced inactivation of L. monocytogenes at concentrations higher than 2.6 M. A low concentration of sodium chloride within 1.7 M had no effect on HHP-induced inactivation. Almost complete inactivation at relatively low sodium chloride concentration solution was observed with treatments above 500 MPa. Sucrose also significantly inhibited HHP-induced inactivation of L. monocytogenes when greater than 1.2 M sucrose solutions were used. HHP-treatment at 400 MPa reduced the number of L. monocytogenes in 1.2 M, 1.5 M, and 1.8 M sucrose solutions by 4.8, 2.0, and 0.7 log cycles, respectively. Higher pressure did not yield significant reductions. Sodium phosphate buffer significantly inhibited HHP-induced inactivation of L. monocytogenes. In particular, 1 M phosphate buffer completely inhibited HHP-induced inactivation even at 600 MPa. HHP-treatment at 400 MPa reduced the number of L. monocytogenes in 0.1 M, 0.25 M, and 0.5 M phosphate buffer solutions by 5.6, 4.1, and 3.2 log cycles, respectively. The effect of HHP-induced inactivation of L. monocytogenes in the three kinds of solution was evaluated by adjusting water activity (a(w)). However, the baroprotective effect differed depending on the kind of solute even at the same a(w). This result showed no consistent correlation between a(w) and solute concentration in terms of the baroprotective effect. As an alternative approach, saturation of suspension solution was used for evaluating the effect of HHP-induced inactivation of L. monocytogenes. As the saturation of suspension media increased, the effect of HHP-induced inactivation of L. monocytogenes decreased regardless of the kinds of solute. The saturation of solution would be an alternative parameter of inhibition in terms of HHP-induced inactivation of bacteria.     

Effect of reheating on viability of a five-strain mixture of Listeria monocytogenes in vacuum-sealed packages of frankfurters following refrigerated or frozen storage

The purpose of this study was to assess consumer preferences for storing and reheating frankfurters and to use this information to assess the effect of product formulation and storage times and temperatures on the viability of Listeria monocytogenes after reheating of frankfurters. Individual links were inoculated with about 8.0 log CFU per package of a five-strain mixture of the pathogen, vacuum sealed, and stored at 4 degrees C for 3 and 15 days and at -18 degrees C for 30 days. Frankfurters formulated with and without 2% added potassium lactate were heated to a surface temperature of 60, 70, 80, or 90 degrees C for up to 8 min by submersing the packages in a thermostatically controlled circulating water bath. Surviving bacteria were recovered and counted by rinsing the contents of each package with sterile peptone water and plating this solution directly onto modified Oxford selective agar plates. In general, the results revealed that about a 5-log unit reduction was achieved by reheating to a surface temperature of 70 degrees C for about 2 min or 80 or 90 degrees C for about 0.6 min regardless of storage conditions or formulation. Product formulation did not appreciably affect the viability of the pathogen after heating; there was no appreciable difference in the number of cells surviving the heat treatment in product prepared with or without potassium lactate. These findings can be used to establish reheating guidelines for consumers to ensure that frankfurters, which may become contaminated with low levels of L. monocytogenes prior to packaging and after unpackaging, are adequately reheated prior to consumption.     

Effectiveness of electrolyzed acidic water in killing Escherichia coli O157:H7, Salmonella enteritidis,and Listeria monocytogenes on the surfaces of tomatoes

A study was conducted to evaluate the efficacy of electrolyzed acidic water, 200-ppm chlorine water, and sterile distilled water in killing Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes on the surfaces of spot-inoculated tomatoes. Inoculated tomatoes were sprayed with electrolyzed acidic water, 200-ppm chlorine water, and sterile distilled water (control) and rubbed by hand for 40 s. Populations of E. coli O157:H7, Salmonella, and L. monocytogenes in the rinse water and in the peptone wash solution were determined. Treatment with 200-ppm chlorine water and electrolyzed acidic water resulted in 4.87- and 7.85-log10 reductions, respectively, in Escherichia coli O157:H7 counts and 4.69- and 7.46-log10 reductions, respectively, in Salmonella counts. Treatment with 200-ppm chlorine water and electrolyzed acidic water reduced the number of L. monocytogenes by 4.76 and 7.54 log10 CFU per tomato, respectively. This study's findings suggest that electrolyzed acidic water could be useful in controlling pathogenic microorganisms on fresh produce.     

Calcinated calcium killing of Escherichia coli O157:H7, Salmonella,and Listeria monocytogenes on the surface of tomatoes

This study was conducted to evaluate the efficacy of calcinated calcium, 200 ppm chlorine water (1% active chlorine), and sterile distilled water in killing Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes on the surfaces of spot-inoculated tomatoes. Inoculated tomatoes were sprayed with calcinated calcium, chlorinated water, or sterile distilled water (control) and hand rubbed for 30 s. Populations of E coli O157:H7, Salmonella, and L. monocytogenes in the rinse water and in the residual (0.1% peptone) wash solution were determined. Treatment with 200 ppm chlorine and calcinated calcium resulted in 3.40- and 7.85-log10 reductions of E. coli O157:H7, respectively, and 2.07- and 7.36-log10 reductions of Salmonella, respectively. Treatment with 200 ppm chlorine and calcinated calcium reduced L monocytogenes numbers by 2.27 and 7.59 log10 CFU per tomato, respectively. The findings of this study suggest that calcinated calcium could be useful in controlling pathogenic microorganisms in fresh produce.     

Comparison of excision and swabbing sampling methods to determine the microbiological quality of swine carcass surfaces

Although different sampling methods are available to determine the microbiological quality of animal carcass surfaces, these vary in their ability to recover bacteria quantitatively. In this study, excision and swabbing (one-site vs three-site) sampling methods were compared for their recovery (both numbers and incidence) of total aerobic bacteria (APC), total coliforms, and Escherichia coli biotype I from swine carcass surfaces. In addition, the effect of refrigerated storage (to simulate shipment to an off-site laboratory) was investigated, as well as the effectiveness of the one vs three anatomical site swabbing methods. Based on samples from 120 market swine carcasses, the excision method recovered the highest levels (P>0·05) of all three groups of bacteria (APCs, coliforms, and E.coli), followed by the three-site method and finally the one-site method. For the APCs, the recovery, given as log cfu cm⁻², were 4·7, 4·0 and 3·3 respectively for excision, three-site swab, and one-site swab methods; for coliforms, the recovery was 2·1, 0·3 and −0·1 respectively; and for E.coli it was 2·4, 0·3, and −0·3 respectively. Comparing the swabbing methods, the three-site swab method recovered higher levels (P>0·05) of the three groups of bacteria than the one-site swab method, with the ham being responsible for this increased recovery. Excision and the three-site swab method gave a higher incidence of E. coli (45 and 58 positive (one colony on one of the duplicate plates) out of 120 carcasses respectively) than the one-site swab method (23 positive out of 120 carcasses). Storage of the swabs overnight in the cold did not cause a decrease (P>0·05) in either the number or incidence of bacteria recovered. These data indicate that while the excision method will recover the highest number and incidence of bacteria, swabbing methods can provide an alternative to this more labor intensive and destructive method.     

Phosphate buffer increases recovery of Escherichia coli O157:H7 from frozen apple juice

It is common practice to dilute food products in 0.1% peptone before microbiological analysis. However, this diluent may not be appropriate for detection of injured organisms present in acidic foods. Shelf-stable unclarified apple juice (pH 3.6) was inoculated with approximately 1 x 10(7) CFU/ml of Escherichia coli O157:H7 and held at 23 +/- 2 degrees C (control) or frozen to -20 +/- 2 degrees C for 24 h to induce injury before sampling. Unfrozen or frozen and thawed juice was diluted 1:1 or 1:10 in 0.1% (wt/vol) peptone (pH 6.1) or 0.1 M phosphate buffer (pH 7.2). Juice samples were plated onto tryptic soy agar with 0.1% (wt/vol) sodium pyruvate (TSAP) to measure survival or onto sorbitol MacConkey agar (SMA) to indicate injury. Counts on TSAP or SMA were the same for control samples held in peptone or phosphate buffer for up to 45 min. However, populations of E. coli in frozen and thawed samples declined rapidly upon dilution in 0.1% peptone. Within 20 min, E. coli underwent a >1-log10 CFU/ml reduction in viability as measured on TSAP and a >2-log10 CFU/ml reduction to below the limit of detection (1.6 or 2.3 log10 CFU/ml) on SMA. In contrast, populations of E. coli in frozen and thawed samples diluted in phosphate buffer did not decrease significantly on TSAP and decreased by <0.6 log CFU/ml on SMA during a 45-min holding period. The acidity of apple juice appears to interfere with the recovery of freeze-thaw-injured E. coli O157:H7 during sampling. Using 0.1 M phosphate buffer (pH 7.2) as a diluent results in superior recovery of these organisms on both selective and nonselective plating media.     

使用不同稀释液对食品中菌落总数检测结果的影响初探

目的:对比将灭菌蒸馏水、生理盐水、磷酸盐缓冲液及缓冲蛋白胨水作为稀释液进行食品菌落总数检测的结果,初步探讨各类稀释液对检测结果的影响。方法:将六大类共90份食品,同时使用不同稀释液进行菌落总数检测,通过其检测数据的配对t检验,分析其相关性并探讨其对检测结果的影响。结果:多数食品使用蒸馏水的检测结果偏低,使用磷酸盐缓冲液和生理盐水的检测结果一致性较高,使用缓冲蛋白胨水检测结果偏高,以磷酸盐缓冲液结果为参照,经配对T检验,使用蒸馏水、生理盐水、缓冲蛋白胨水结果的P值分别为:0.000、0.166、0.059,仅使用蒸馏水的数据存在显著性差异。结论:使用蒸馏水在多数情况下(高盐高糖等食品除外)不利于菌落总数的准确检验,而使用生理盐水、磷酸盐缓冲液、缓冲蛋白胨水结果无显著性差异。     

Efficacy of electrolyzed water in the inactivation of planktonic and biofilm Listeria monocytogenes in the presence of organic matter

The ability of electrolyzed (EO) water to inactivate Listeria monocytogenes in suspension and biofilms on stainless steel in the presence of organic matter (sterile filtered chicken serum) was investigated. A five-strain mixture of L. monocytogenes was treated with deionized, alkaline EO, and acidic EO water containing chicken serum (0, 5, and 10 ml/liter) for 1 and 5 min. Coupons containing L. monocytogenes biofilms were also overlaid with chicken serum (0, 2.5, 5.0, and 7.5 ml/liter) and then treated with deionized water, alkaline EO water, acidic EO water, alkaline EO water followed by acidic EO water, and a sodium hypochlorite solution for 30 and 60 s. Chicken serum decreased the oxidation-reduction potential and chlorine concentration of acidic EO water but did not significantly affect its pH. In the absence of serum, acidic EO water containing chlorine at a concentration of 44 mg/liter produced a > 6-log reduction in L. monocytogenes in suspension, but its bactericidal activity decreased with increasing serum concentration. Acidic EO water and acidified sodium hypochlorite solution inactivated L. monocytogenes biofilms to similar levels, and their bactericidal effect decreased with increasing serum concentration and increased with increasing time of exposure. The sequential 30-s treatment of alkaline EO water followed by acidic EO water produced 4- to 5-log reductions in L. monocytogenes biofilms, even in the presence of organic matter.