Fate of Listeria monocytogenes during freezing, thawing and home storage of frankfurters.

Little information is available regarding the fate of Listeria monocytogenes during freezing, thawing and home storage of frankfurters even though recent surveys show that consumers regularly store unopened packages in home freezers. This study examined the effects of antimicrobials, refrigerated storage, freezing, thawing method, and post-thawing storage (7 degrees C) on L. monocytogenes on frankfurters. Inoculated (2.1 log CFU/cm(2)) frankfurters formulated without (control) or with antimicrobials (1.5% potassium lactate plus 0.1% sodium diacetate) were vacuum-packaged, stored at 4 degrees C for 6 or 30 d and then frozen (-15 degrees C) for 10, 30, or 50 d. Packages were thawed under refrigeration (7 degrees C, 24 h), on a countertop (23 +/- 2 degrees C, 8 h), or in a microwave oven (2450 MHz, 1100 watts, 220 s followed by 120 s holding), and then stored aerobically (7 degrees C) for 14 d. Bacterial populations were enumerated on PALCAM agar and tryptic soy agar plus 0.6% yeast extract. Antimicrobials completely inhibited (p < 0.05) growth of L. monocytogenes at 4 degrees C for 30 d under vacuum-packaged conditions, and during post-thawing aerobic storage at 7 degrees C for 14 d. Different intervals between inoculation and freezing (6 or 30 d) resulted in different pathogen levels on control frankfurters (2.1 or 3.9 log CFU/cm(2), respectively), while freezing reduced counts by <1.0 log CFU/cm(2). Thawing treatments had little effect on L. monocytogenes populations (<0.5 log CFU/cm(2)), and post-thawing fate of L. monocytogenes was not influenced by freezing or by thawing method. Pathogen counts on control samples increased by 1.5 log CFU/cm(2) at d-7 of aerobic storage, and reached 5.6 log CFU/cm(2) at d-14. As indicated by these results, consumers should freeze frankfurters immediately after purchase, and discard frankfurters formulated without antimicrobials within 3 d of thawing and/or opening.

Evaluation of changes in Listeria monocytogenes populations on frankfurters at different stages from manufacturing to consumption.

This study evaluated the fate of inoculated Listeria monocytogenes on frankfurters stored under conditions simulating those that may be encountered between manufacturing and consumption. Frankfurters with or without 1.5% potassium lactate and 0.1% sodium diacetate (PL/SD) were inoculated (1.8 +/- 0.1 log CFU/cm(2)) with a 10-strain composite of L. monocytogenes, vacuum-packaged, and stored under conditions simulating predistribution storage (24 h, 4 degrees C), temperature abuse during transportation (7 h, 7 degrees C followed by 7 h, 12 degrees C), and storage before purchase (60 d, 4 degrees C; SBP). At 0, 20, 40, and 60 d of SBP, samples were exposed to conditions simulating delivery from stores to homes or food establishments (3 h, 23 degrees C), and then opened or held vacuum-packaged at 4 or 7 degrees C for 14 d (SHF). Pathogen counts remained relatively constant on frankfurters with PL/SD regardless of product age and storage conditions; however, they increased on product without antimicrobials. In vacuum-packaged samples, during SHF at 4 degrees C, the pathogen grew faster (P < 0.05) on older product (20 d of SBP) compared to product that was fresh (0 d of SBP); a similar trend was observed in opened packages. At 7 degrees C, the fastest growth (0.35 +/- 0.02 log CFU/cm(2)/d) was observed on fresh product in opened packages; in vacuum-packages, growth rates on fresh and aged products were similar. By day 40 of SBP the pathogen reached high numbers and increased slowly or remained unchanged during SHF. This information may be valuable in L. monocytogenes risk assessments and in development of guidelines for storage of frankfurters between package opening and product consumption.

Effect of boiling water carcass immersion on aerobic bacteria counts of poultry skin and processed ground poultry meat.

This study was conducted to determine the relationship between bacteria destruction on poultry carcass skin and bacteria in raw ground poultry meat from the same carcasses. Immersion time in boiling water of broiler chicken whole carcasses required for maximum reduction of naturally occurring aerobic bacterial count on skin was measured. Treatments for chicken carcasses consisted of immersion in boiling water (approximately 95 degrees C) for 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, and 4 min. Four skin samples taken following treatment and three taken from subsequently ground carcass meat were analyzed for total aerobic plate counts (APC). Analysis of the data indicated a linear increase in bacterial destruction on skin with increased boiling water immersion time from 0 to 4 min. Reduction of skin bacteria to less than 1 log10 occurred at 3 min carcass immersion or longer. The analysis also indicated that treatment with boiling water and removal of skin was effective in reducing bacterial counts in ground meat to similar levels at all treatment times from 0.5 to 4.0 min. Findings from this study indicated that a boiling water immersion intervention and removal of skin could reduce subsequent bacteria contamination of ground meat. This intervention could minimize the risk of pathogen-contaminated primary processed poultry carcasses used in further processing.

Modeling the effect of inoculum size and acid adaptation on growth/no growth interface of Escherichia coli O157:H7.

The objective of this study was to model with logistic regression the growth/no growth interface of different initial inoculation levels (10(1), 10(3) and 10(5) CFU/ml; study 1), or nonadapted vs acid-adapted (study 2) Escherichia coli O157:H7 as influenced by pH, NaCl concentration and incubation temperature. Study 1 was conducted with a mixture of four E. coli O157:H7 strains grown (35 degrees C, 24 h) in tryptic soy broth (TSB). Study 2 was conducted with the same mixture of four E. coli O157:H7 strains grown (35 degrees C, 24 h) in glucose-free TSB with 1% added glucose (final pH 4.83), or in diluted lactic acid meat decontamination runoff fluids (washings; final pH 4.92), or nonadapted cultures prepared in glucose-free TSB (final pH 6.45), or in water washings (final pH 6.87). Parameters included incubation temperature (10-35 degrees C), pH (3.52-7.32), and NaCl concentration (0-10% w/v). Growth responses were evaluated for 60 days turbidimetrically (610 nm) every 5 days in 160 (study 1) and 360 (study 2) combinations in quadruplicate samples, with a microplate reader. The lower the initial inoculum the higher were the minimum pH and a(w) values permitting growth. Differences in the pH and a(w) growth limits among inoculum concentrations increased at 15 and 10 degrees C. Acid-adapted cultures were able to grow at lower pH than nonadapted cultures, while at temperatures below 25 degrees C, growth initiation of nonadapted cultures stopped at higher a(w) compared to acid-adapted cultures for the whole pH range of 3.52 to 7.32. A comparison with available data indicated that our model for acid-adapted E. coli O157:H7 in different environments may provide representative growth probabilities covering both nonadapted and stress-adapted contaminants.

Control of Listeria monocytogenes on frankfurters with antimicrobials in the formulation and by dipping in organic acid solutions.

The antilisterial activity of sodium lactate (SL) and sodium diacetate (SD) was evaluated in a frankfurter formulation and in combination with a dipping treatment into solutions of lactic acid or acetic acid after processing and inoculation. Pork frankfurters were formulated with 1.8% SL or 0.25% SD or combinations of 1.8% SL with 0.25 or 0.125% SD. After processing, frankfurters were inoculated (2 to 3 log CFU/cm2) with a 10-strain composite of Listeria monocytogenes and left undipped or were dipped (2 min) in 2.5% solutions of lactic acid or acetic acid (23 +/- 2 degrees C) before vacuum packaging and storage at 10 degrees C for 40 days. Total microbial populations and L. monocytogenes, lactic acid bacteria, and yeasts and molds were enumerated during storage. Sensory evaluations also were carried out on frankfurters treated and/or formulated with effective antimicrobials. The combination of 1.8% SL with 0.25% SD provided complete inhibition of L. monocytogenes growth throughout storage. Dipping in lactic acid or acetic acid reduced initial populations by 0.7 to 2.1 log CFU/cm2, but during storage (12 to 20 days), populations on dipped samples without antimicrobials in the formulation reached 5.5 to 7.9 log CFU/cm2. For samples containing single antimicrobials and dipped in lactic acid or acetic acid, L. monocytogenes growth was completely inhibited or reduced over 12 and 28 days, respectively, whereas final populations were lower (P < 0.05) than those in undipped samples of the same formulations. Bactericidal effects during storage (reductions of 0.6 to 1.0 log CFU/ cm2 over 28 to 40 days) were observed in frankfurters containing combinations of SL and SD that were dipped in organic acid solutions. Inclusion of antimicrobials in the formulation and/or dipping the product into organic acid solutions did not affect (P > 0.05) the flavor and overall acceptability of products compared with controls. The results of this study may be valuable to meat processors as they seek approaches for meeting new regulatory requirements in the United States.

Effect of simulated spray chilling with chemical solutions on acid-habituated and non-acid-habituated Escherichia coli O157:H7 cells attached to beef carcass tissue.

Samples (10 by 20 by 2.5 cm) of beef carcass tissue were inoculated (10(4) to 10(5) CFU/cm2) with Escherichia coli O157: H7 that was either non-acid habituated (prepared by incubating at 15 degrees C for 48 h in inoculated filter-sterilized composite [1:1] of hot and cold water meat decontamination runoff fluids, pH 6.05) or acid habituated (prepared in inoculated water fluids mixed with filter-sterilized 2% lactic acid [LA] runoff fluids in a proportion of 1/99 [vol/vol], pH 4.12). The inoculated surfaces were exposed to conditions simulating carcass chilling (- 3 degrees C for 10 h followed by 38 h at 1 degree C). Treatments applied to samples (between 0 and 10 h) during chilling included the following: (i) no spraying (NT) or spraying (for 30 s every 30 min) with (ii) water, (iii) cetylpyridinium chloride (CPC; 0.1 or 0.5%), (iv) ammonium hydroxide (AH; 0.05%), (v) lactic acid (LA; 2%), (vi) acidified sodium chlorite (ASC; 0.12%), (vii) peroxyacetic acid (PAA; 0.02%), (viii) sodium hydroxide (SH; 0.01%), or (ix) sodium hypochlorite (SC; 0.005%) solutions of 4 degrees C. Samples were taken at 0, 10, 24, 36, and 48 h of the chilling process to determine changes in E. coli O157:H7 populations. Phase 1 tested water, SH, PAA, LA, and 0.5% CPC on meat inoculated with non-acid-habituated pathogen populations, whereas phase 2 tested water, SC, AH, ASC, LA, and 0.1% CPC on meat inoculated with acid- and non-acid-habituated populations. Reductions in non-acid-habituated E. coli O157:H7 populations from phase 1 increased in the order NT = water = SH < PAA < LA < CPC. Reductions from phase 2 for acid-habituated cells increased in the order NT = water = SC < ASC = LA = AH < CPC, whereas on non-acid-habituated cells the order observed was NT = water = SC < AH = ASC < LA < CPC. Previous acid habituation of E. coli O157:H7 inocula rendered the cells more resistant to the effects of spray chilling, especially with acid; however, the trend of reduction remained spray chilling with water = non-spray chilling < spray chilling with chemical solutions.

Thermal inactivation of susceptible and multiantimicrobial-resistant salmonella strains grown in the absence or presence of glucose.

The heat resistance of susceptible and multiantimicrobial-resistant Salmonella strains grown to stationary phase in glucose-free tryptic soy broth supplemented with 0.6% yeast extract (TSBYE-G; nonadapted), in regular (0.25% glucose) TSBYE, or in TSBYE-G with 1.00% added glucose (TSBYE+G; acid adapted) was determined at 55, 57, 59, and 61 degrees C. Cultures were heated in sterile 0.1% buffered peptone water (50 microl) in heat-sealed capillary tubes immersed in a thermostatically controlled circulating-water bath. Decimal reduction times (D values) were calculated from survival curves having r(2) values of >0.90 as a means of comparing thermal tolerance among variables. D(59 degrees C) values increased (P < 0.05) from 0.50 to 0.58 to 0.66 min for TSBYE-G, TSBYE, and TSBYE+G cultures, respectively. D(61 degrees C) values of antimicrobial-susceptible Salmonella strains increased (P < 0.05) from 0.14 to 0.19 as the glucose concentration increased from 0.00 to 1.00%, respectively, while D(61 degrees C) values of multiantimicrobial-resistant Salmonella strains did not differ (P > 0.05) between TSBYE-G and TSBYE+G cultures. When averaged across glucose levels and temperatures, there were no differences (P > 0.05) between the D values of susceptible and multiantimicrobial-resistant inocula. Collectively, D values ranged from 4.23 to 5.39, 1.47 to 1.81, 0.50 to 0.66, and 0.16 to 0.20 min for Salmonella strains inactivated at 55, 57, 59, and 61 degrees C, respectively. z(D) values were 1.20, 1.48, and 1.49 degrees C for Salmonella strains grown in TSBYE+G, TSBYE, and TSBYE-G, respectively, while the corresponding activation energies of inactivation were 497, 493, and 494 kJ/mol. Study results suggested a cross-protective effect of acid adaptation on thermal inactivation but no association between antimicrobial susceptibility and the ability of salmonellae to survive heat stress.

Acid adaptation does not promote survival or growth of Listeria monocytogenes on fresh beef following acid and nonacid decontamination treatments.

The objective of this study was to evaluate the survival and growth of acid-adapted and nonadapted Listeria monocytogenes inoculated onto fresh beef subsequently treated with acid or nonacid solutions. Beef slices (2.5 by 5 by 1 cm) from top rounds were inoculated with acid-adapted or nonadapted L. monocytogenes (4.6 to 5.0 log CFU/cm2) and either left untreated (control) or dipped for 30 s in water at 55 degrees C, water at 75 degrees C, 2% lactic acid at 55 degrees C, or 2% acetic acid at 55 degrees C. The beef slices were vacuum packaged and stored at 4 or 10 degrees C and were analyzed after 0, 7, 14, 21, and 28 days of storage. Dipping in 75 degrees C water, lactic acid, and acetic acid resulted in immediate pathogen reductions of 1.4 to 2.0, 1.8 to 2.6, and 1.4 to 2.4 log CFU/cm2, respectively. After storage at 10 degrees C for 28 days, populations of L. monocytogenes on meat treated with 55 degrees C water increased by ca. 1.6 to 1.8 log CFU/cm2. The pathogen remained at low population levels (1.6 to 2.8 log CFU/cm2) on acid-treated meat, whereas populations on meat treated with 75 degrees C water increased rapidly, reaching levels of 3.6 to 4.6 log CFU/cm2 by day 14. During storage at 4 degrees C, there was no growth of the pathogen for at least 21 days in samples treated with 55 and 75 degrees C water, and periods of no growth were longer for acid-treated samples. There were no differences between acid-adapted and nonadapted organisms across treatments with respect to survival or growth. In conclusion, the dipping of meat inoculated with L. monocytogenes into acid solutions reduced and then inhibited the growth of the pathogen during storage at 4 and 10 degrees C, while dipping in hot water allowed growth despite initial reductions in pathogen contamination. The results of this study indicate a residual activity of acid-based decontamination treatments compared with water-based treatments for refrigerated (4 degrees C) or temperature-abused (10 degrees C) lean beef tissue in vacuum packages, and these results also indicate that this activity may not be counteracted by prior acid adaptation of L. monocytogenes.

Comparative analysis of acid resistance between susceptible and multi-antimicrobial-resistant Salmonella strains cultured under stationary-phase acid tolerance-inducing and noninducing conditions.

This study compared acid resistance levels among five antimicrobial-susceptible strains of Salmonella and five strains that were simultaneously resistant to a minimum of six antimicrobial agents. The induction of a stationary-phase acid tolerance response (ATR) was attempted by both transient low-pH acid shock and acid adaptation. For acid shock induction, strains were grown for 18 h in minimal E medium containing 0.4% glucose (EG medium) and exposed to sublethal acid stress (pH 4.3) for 2 h, and subsequently, both shocked and nonshocked cultures were acid challenged (pH 3.0) for 4 h. Acid adaptation was achieved by growing strains for 18 h in tryptic soy broth containing 1.0% glucose (TSB+G), while nonadapted cultures were grown for 18 h in glucose-free tryptic soy broth (TSB-G). Acid-adapted and nonadapted inocula were acid challenged (pH 2.3) for 4 h. Initial (0 h) mean populations of nonchallenged Salmonella were 8.5 to 8.7, 8.4 to 8.8, and 8.2 to 8.3 log CFU/ml for strains grown in EG medium, TSB-G, and TSB+G, respectively. After 4 h of acid challenge, mean populations were 3.0 to 4.8 and 2.5 to 3.7 log CFU/ml for previously acid-shocked susceptible and resistant strains, respectively, while corresponding counts for nonshocked strains were 4.3 to 5.5 log CFU/ml and 3.9 to 4.9 log CFU/ml. Following 4 h of acid exposure, acid-adapted cultures of susceptible and resistant strains had mean populations of 6.1 to 6.4 log CFU/ml and 6.4 to 6.6 log CFU/ml, respectively, while corresponding counts for nonadapted cultures were 1.9 to 2.1 log CFU/ml and 1.8 to 2.0 log CFU/ml, respectively. A low-pH-inducible ATR was not achieved through transient acid shock, while an ATR was evident following acid adaptation, as adapted populations were 4.2 to 4.8 log units larger than nonadapted populations following acid exposure. Although some strain-dependent variations in acid resistance were observed, results from this study suggest no association between susceptibility to antimicrobial agents and the ability of the Salmonella strains evaluated to survive low-pH stress.

Biofilm formation by acid-adapted and nonadapted Listeria monocytogenes in fresh beef decontamination washings and its subsequent inactivation with sanitizers.

The antimicrobial effects of sodium hypochlorite (SH, 200 ppm, at an adjusted pH of 6.80 +/- 0.20 and at an unadjusted pH of 10.35 +/- 0.25), quaternary ammonium compound (pH 10.20 +/- 0.12, 200 ppm), and peroxyacetic acid (PAA, pH 3.45 +/- 0.20, 150 ppm) on previously acid-adapted or nonadapted Listeria monocytogenes inoculated (10(5) CFU/ml) into beef decontamination water washings were evaluated. The effects of the sanitizers on suspended cells (planktonic or deattached) and on cells attached to stainless steel coupons obtained from inoculated washings stored at 15 degrees C for up to 14 days were studied. Cells were exposed to sanitizers on days 2, 7, and 14. The pathogen had formed a biofilm of 5.3 log CFU/cm2 by day 2 of storage (which was reduced to 4.6 log CFU/cm2 by day 14), while the total microbial populations showed more extensive attachment (6.1 to 6.6 log CFU/cm2). The sanitizers were more effective in reducing populations of cells in suspension than in reducing populations of attached cells. Overall, there were no differences between previously acid-adapted and nonadapted L monocytogenes with regard to sensitivity to sanitizers. The total microbial biofilms were the most sensitive to all of the sanitizers on day 2, but their resistance increased during storage, and they were at their most resistant on day 14. Listeria monocytogenes displayed stronger resistance to the effects of the sanitizers on day 7 than on day 2 but had become sensitized to all sanitizers by day 14. SH at the adjusted pH (6.80) (ASH) was generally more effective in reducing bacterial populations than was SH at the unadjusted pH. PAA generally killed attached cells faster at 30 to 300 s of exposure than did the other sanitizers, except for ASH on day 2. PAA was more effective in killing attached cells than in killing cells treated in suspension, in contrast to the other sanitizers.

In vitro expansion of cord blood does not prevent engraftment of severe combined immunodeficient repopulating cells.

This study aimed to assess the potential of human cord blood (CB) cells to engraft in the xenogenic non-obese diabetic/severe combined immunodeficient (NOD/SCID) mouse model after in vitro expansion culture. We also studied the quality of human haemopoiesis arising from the transplantation of fresh or expanded cells in this model. Cord blood CD34(+) cells were cultured for 3, 7 or 10 d with stem cell factor, Flt3, thrombopoietin, interleukin 3 (IL-3), IL-6 and granulocyte colony-stimulating factor, all at 10 ng/ml in serum-replete conditions. Transplantation of mice with fresh CB containing 3 x 10(4) CD34(+) cells and 1-2 SCID repopulating cells (SRC) resulted in a median of 7.4% (0.4%-76.8%) human engraftment. When mice received the expanded product of 1-2 SRC, the ability to repopulate NOD/SCID mice was maintained even after 10 d of in vitro culture. Serial dilution of the expanded cells suggested that in vitro expansion had increased SRC numbers two- to fourfold. Expanded SRC produced long-term culture-initiating cells, clonogenic cells and CD34(+) cells in the same proportions as fresh cells after successful engraftment. Therefore, expanded SRC were able to differentiate in the same way as fresh SRC. There was a trend towards lower levels of engraftment when d 7 cultured cells were transplanted (median engraftment 0.8%, range 0.0-24.0%) compared with 1-2 fresh SRC. Our data suggest that this is owing to reduced proliferation of cultured cells in vivo. By utilizing limiting numbers of CB SRC, we confirmed that the engraftment potential of SRC in the NOD/SCID model was preserved after in vitro expansion. Furthermore, dilution experiments strongly suggest two- to fourfold expansion of SRC in vitro. These studies are relevant for developing clinical stem cell expansion strategies.

Exposure to non-acid fresh meat decontamination washing fluids sensitizes Escherichia coli O157:H7 to organic acids.

AIMS: To investigate whether Escherichia coli O157:H7 maintains acid tolerance in water meat decontamination washing fluids. METHODS AND RESULTS: A rifampicin-resistant derivative of E. coli O157:H7 strain ATCC 43895 was inoculated (10(5) cfu ml(-1)) in spray-washings from meat sprayed with cold (10 degrees C) or hot (85 degrees C) water, stored at 10 degrees C for up to 14 days, and its acid tolerance was assessed at 2 and 8 days by exposure to broth or new washings adjusted to pH 3.5 or 3.7 with lactic or acetic acid. The pathogen survived in the water washings, but it was outgrown by the natural, Pseudomonas-like flora, and it was sensitized to acid. CONCLUSIONS: The acid tolerance of E. coli O157:H7 decreases following exposure to non-acid, but otherwise stressful, conditions prevailing in water meat washings at 10 degrees C. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings suggest that the more intense use of water-based technologies should be included in meat decontamination strategies because they may contribute to enhanced meat safety by inducing acid sensitization in E. coli O157:H7.

Ascorbic acid enhances destruction of Escherichia coli O157:H7 during home-type drying of apple slices.

Destruction of Escherichia coli O157:H7 was evaluated on inoculated apple slices dehydrated at two temperatures with and without application of predrying treatments. Half-ring slices (0.6 cm thick) of peeled and cored Gala apples were inoculated by immersion for 30 min in a four-strain composite inoculum of E. coli O157:H7. The inoculated slices (8.7 to 9.4 log CFU/g) either received no predrying treatment (control), were soaked for 15 min in a 3.4% ascorbic acid solution, or were steam blanched for 3 min at 88 degrees C immediately prior to drying at 57.2 or 62.8 degrees C for up to 6 h. Samples were plated on tryptic soy (TSA) and sorbitol MacConkey (SMAC) agar media for direct enumeration of surviving bacterial populations. Steam blanching changed initial inoculation levels by +0.3 to -0.7 log CFU/g, while immersion in the ascorbic acid solution reduced the inoculation levels by 1.4 to 1.6 log CFU/g. Dehydration of control samples for 6 h reduced mean bacterial populations by 2.9 log CFU/g (TSA or SMAC) at 57.2 degrees C and by 3.3 (SMAC) and 3.5 (TSA) log CFU/g at 62.8 degrees C. Mean decreases from initial inoculum levels for steam-blanched slices after 6 h of drying were 2.1 (SMAC) and 2.0 (TSA) log CFU/g at 57.2 degrees C, and 3.6 (TSA or SMAC) log CFU/g at 62.8 degrees C. In contrast, initial bacterial populations on ascorbic acid-pretreated apple slices declined by 5.0 (SMAC) and 5.1 (TSA) log CFU/g after 3 h of dehydration at 57.2 degrees C, and by 7.3 (SMAC) and 6.9 (TSA) log CFU/g after 3 h at 62.8 degrees C. Reductions on slices treated with ascorbic acid were in the range of 8.0 to 8.3 log CFU/g after 6 h of drying, irrespective of drying temperature or agar medium used. The results of immersing apple slices in a 3.4% ascorbic acid solution for 15 min prior to drying indicate that a predrying treatment enhances the destruction of E. coli O157:H7 on home-dried apple products.

Fate of Escherichia coli O157:H7, Salmonella typhimurium DT 104, and Listeria monocytogenes in fresh meat decontamination fluids at 4 and 10 degrees C.

Bacterial pathogens may colonize meat plants and increase food safety risks following survival, stress hardening, or proliferation in meat decontamination fluids (washings). The objective of this study was to evaluate the ability of Escherichia coli O157:H7, Salmonella Typhimurium DT 104, and Listeria monocytogenes to survive or grow in spray-washing fluids from fresh beef top rounds sprayed with water (10 or 85 degrees C) or acid solutions (2% lactic or acetic acid, 55 degrees C) during storage of the washings at 4 or 10 degrees C in air to simulate plant conditions. Inoculated Salmonella Typhimurium DT 104 (5.4 +/- 0.1 log CFU/ml) died off in lactate (pH 2.4 +/- 0.1) and acetate (pH 3.1 +/- 0.2) washings by 2 days at either storage temperature. In contrast, inoculated E. coli O157:H7 (5.2 +/- 0.1 log CFU/ml) and L. monocytogenes (5.4 +/- 0.1 log CFU/ml) survived in lactate washings for at least 2 days and in acetate washings for at least 7 and 4 days, respectively; their survival was better in acidic washings stored at 4 degrees C than at 10 degrees C. All inoculated pathogens survived in nonacid (pH > 6.0) washings, but their fate was different. E. coli O157:H7 did not grow at either temperature in water washings, whereas Salmonella Typhimurium DT 104 failed to multiply at 4 degrees C but increased by approximately 2 logs at 10 degrees C. L. monocytogenes multiplied (0.6 to 1.3 logs) at both temperatures in water washings. These results indicated that bacterial pathogens may survive for several days in acidic, and proliferate in water, washings of meat, serving as potential cross-contamination sources, if pathogen niches are established in the plant. The responses of surviving pathogens in meat decontamination waste fluids to acid or other stresses need to be addressed to better evaluate potential food safety risks.

Influence of the natural microbial flora on the acid tolerance response of Listeria monocytogenes in a model system of fresh meat decontamination fluids.

Depending on its composition and metabolic activity, the natural flora that may be established in a meat plant environment can affect the survival, growth, and acid tolerance response (ATR) of bacterial pathogens present in the same niche. To investigate this hypothesis, changes in populations and ATR of inoculated (10(5) CFU/ml) Listeria monocytogenes were evaluated at 35 degrees C in water (10 or 85 degrees C) or acidic (2% lactic or acetic acid) washings of beef with or without prior filter sterilization. The model experiments were performed at 35 degrees C rather than lower (8.0 log CFU/ml) by day 1. The pH of inoculated water washings decreased or increased depending on absence or presence of natural flora, respectively. These microbial and pH changes modulated the ATR of L. monocytogenes at 35 degrees C. In filter-sterilized water washings, inoculated L. monocytogenes increased its ATR by at least 1.0 log CFU/ml from days 1 to 8, while in unfiltered water washings the pathogen was acid tolerant at day 1 (0.3 to 1.4 log CFU/ml reduction) and became acid sensitive (3.0 to >5.0 log CFU/ml reduction) at day 8. These results suggest that the predominant gram-negative flora of an aerobic fresh meat plant environment may sensitize bacterial pathogens to acid.

A prospective, physician self-reported adverse incident audit on a general medical unit.

There is some evidence that physician self-reporting is an efficient and effective way of collecting data on adverse incidents in health care. This study tested a simple prospective adverse incident audit, self-reported by physicians, on a general medical unit. A total of 158 reports were collected over a 6-month period covering a wide range of quality issues, including, but not limited to, safety issues. One-third of reported incidents occurred within 48 h of hospitalization. One-half of incidents were associated with harm or inconvenience to patients. Reported incidents fell into easily classifiable groups, and the data was used as a platform for a coordinated approach to quality improvement within the department. It is concluded that this technique is an easily implementable addition to the more traditional methods used for quality improvement within general medicine.

Food safety education: what should we be teaching to consumers?

Food safety education is most effective when messages are targeted toward changing behaviors most likely to result in foodborne illness. The five major control factors for pathogens are personal hygiene, adequate cooking, avoiding cross-contamination, keeping food at safe temperatures, and avoiding foods from unsafe sources. Pathogens associated with poor personal hygiene have the highest incidence and costs. Inadequate cooking and cross-contamination have lower incidence. Keeping food at safe temperatures and unsafe food sources have the lowest incidence, although costs per case are sometimes very high. We recommend that consumer food safety educators primarily focus on hand washing, adequate cooking, and avoiding cross-contamination. Secondary messages should focus on keeping food at safe temperatures and avoiding food from an unsafe source. Evaluation tools are needed to evaluate self-reported behavior changes. The evaluation questions must focus on salient behaviors that are most likely to result in foodborne illnesses and must withstand rigorous standards of reliability and validity.

Expanded health belief model predicts diabetes self-management in college students.

An instrument was designed to determine relationships between constructs of the Expanded Health Belief Model and to identify characteristics of college students who successfully manage their diabetes. The Diabetes College Scale was developed to measure attitudes and behaviors pertinent to diabetes management and college life. It was tested for content validity, test-retest reliability, and internal consistency. Data were collected from college students using a cross-sectional design. Campus health care providers were invited via electronic mail to administer the survey to students with Type I diabetes. Ninety-eight questionnaires were mailed to interested providers, of which 86 (88%) were returned. Mean scores for attitude constructs, seven behaviors, and two outcomes were measured. Twenty-six experts established content validity. Instrument reliability was evaluated using paired t-tests, Cronbach's alpha, and correlation coefficients. Correlation coefficients and stepwise multiple regression analysis evaluated relationships among variables measured. Intention and emotional response were strong predictors of exercise, whereas health importance and intention were predictive of testing blood sugar. Situational factors and emotional response were substantial barriers to optimal diabetes self-care. College health care providers should address these areas in providing services to this population. Additional testing of the instrument is also recommended.

Development and evaluation of an intervention program: "Control on Campus".

PURPOSE: This study was designed to develop and test an intervention for college students with type 1 diabetes. METHODS: A diabetes program, "Control on Campus," and guide were developed based on the Expanded Health Belief Model and Social Learning Theory. Diabetes knowledge, attitudes, and behaviors were assessed preprogram, postprogram, and at follow-up for 3 intervention cohorts and a control group. RESULTS: Reporting of HbA1c values and diabetes knowledge improved significantly as a result of the intervention compared with no increase in the control group. Furthermore, participants reported feeling more support on campus after the intervention, appeared to have overcome their fears associated with testing their blood glucose, reported an increased frequency of blood glucose testing, and were more likely to test when they felt their blood glucose level was low. CONCLUSIONS: Overall, this research yielded substantial insight into the characteristics of college students with diabetes and was successful in designing and evaluating an intervention trial for this population.

Different behaviour of fresh and cultured CD34+ cells during immunomagnetic separation.

In-vitro expansion of human cord blood (CB) cells could enhance peripheral blood recovery and ensure long-term engraftment of larger recipients in the clinical transplant setting. Enrichment of CD34+ cells using the MiniMACS column has been evaluated for the preparation of CB CD34+ cells before and after expansion culture. Repurification of CD34+ cells after culture would assist accurate phenotypic and functional analysis. When fresh CB mononuclear cells (MNC) were separated, the MACS positive (CD34+) fraction (90.1% pure) contained a mean (+/- SD, n = 5) of 93.0 +/- 8.0% of the eluted CD34+ cells, 99.6 +/- 0.7% of the CFU-GM and all of the eluted long-term culture-initiating cells (LTC-IC). Cord blood CD34+ cells were then cultured for 14 d with IL-3, IL-6, SCF, G-CSF and GM-CSF, each at 10 ng/ml. The total cell expansion was 2490 +/- 200-fold and the CD34+ cell expansion was 49 +/- 17-fold. The percentage of CD34+ cells present after expansion culture was 1.2 +/- 0.85%. When these cells were repurified on the MiniMACS column, the MACS positive fraction only contained 40.3 +/- 13.4% of the eluted CD34+ cells which was enriched for the mature CD34+ CD38+ subset, 24.4 +/- 8.8% of the eluted CFU-GM and 79.5 +/- 11.0% of the LTC-IC. The remaining cells were eluted in the MACS negative fraction. In conclusion, repurification of cultured CD34+ cells does not yield a representative population and many progenitors are lost in the MACS negative fraction. This can give misleading phenotypic and functional data. Cell losses may be important in the clinical setting if cultured cells were repurified for purging.