Influence of transportation stress and animal temperament on fecal shedding of Escherichia coli O157:H7 in feedlot cattle.

To test the influence of transportation stress and temperament on shedding of Escherichia coli O157:H7, cattle (n=150) were classified at various stages of production as Excitable, Intermediate or Calm based on a variety of disposition scores. Presence of E. coli O157:H7 was determined by rectal swabs from live animals and from colons collected postmortem. Percentage of cattle shedding E. coli O157:H7 at arrival at the feedlot was approximately equal among temperament groups. Before shipment to the processing facility, a higher (P=0.03) proportion of cattle from the Calm group shed E. coli O157:H7 compared to the other temperament groups. When pooled across all sampling periods, cattle from the Calm group had a greater percentage test positive for E. coli O157:H7. Neither the acute stressor of transportation nor a more excitable temperament led to increased shedding of E. coli O157:H7 in cattle.

Evaluation of nonpathogenic surrogate bacteria as process validation indicators for Salmonella enterica for selected antimicrobial treatments, cold storage, and fermentation in meat.

Prerigor lean and adipose beef carcass tissues were artificially inoculated individually with stationary-phase cultures of five nonpathogenic Escherichia coli cultures that had been previously identified as surrogates for E. coli O157:H7 or a mixture of five Salmonella strains in a fecal inoculum. Each tissue sample was processed with microbial interventions comparable with those used in the meat industry. The log reductions of the E. coli isolates were generally not statistically different from the salmonellae inoculum within a specific treatment. Inoculation experiments were also conducted with ground beef stored at either 4 or -20 degrees C. When compared with the Salmonella inoculum, at least three of the five E. coli strains survived in a manner that was not statistically different from the salmonellae. The E. coli strains and the Salmonella mixed culture were also inoculated into summer sausage batter, and the population enumerated both before and after fermentation. Four of the E. coli strains showed a lower population reduction (higher survival) than the Salmonella mixed culture. The five nonpathogenic E. coli strains may be used as individually or collectively for specific process validation indicators for Salmonella.

Effect of xylitol on adhesion of Salmonella Typhimurium and Escherichia coli O157:H7 to beef carcass surfaces.

Effects of 10% xylitol (a five-carbon sugar alcohol) on adhesion of Escherichia coli O157:H7 and Salmonella Typhimurium to meat surfaces were examined with three approaches. First, beef outside round was inoculated with rifampin-resistant E. coli O157:H7 and Salmonella Typhimurium dispersed in xylitol or peptone solution. Samples were rinsed with water or not rinsed in a 2 x 2 factorial arrangement. No interaction existed between inoculum and rinsing treatments (P > 0.84). Incubation in xylitol had minimal impact on pathogen adhesion (P > 0.76); however, rinsing reduced pathogen cell counts (P < 0.01). Second, meat samples were treated with water, xylitol, or no rinse; inoculated with pathogens dispersed in peptone solution (8.6 log CFU/ml for each pathogen); and then treated with water, xylitol, or no rinse in a 3 x 3 factorial arrangement. No interactions were observed (P > 0.50). Postinoculation rinsing reduced pathogen loads (P < 0.01) without difference between water and xylitol (P > 0.64). Third, carcass surfaces inoculated with pathogens (5.5 log CFU/cm2) were treated with 35 degrees C water wash, 2.5% L-lactic acid spray, 10% xylitol spray, lactic acid plus xylitol, or hot water plus xylitol. Lactic acid treatments reduced Salmonella Typhimurium at 0 h (P < 0.01) and 24 h (P < 0.02). Hot water treatments tended to reduce Salmonella Typhimurium at 0 h (P < 0.07). Xylitol did not reduce pathogens (P > 0.62) or increase effectiveness of other treatments. Xylitol does not influence E. coli O157:H7 and Salmonella Typhimurium adhesion to meat surfaces.

Biomechanical and microbiological changes in natural hog casings treated with ozone.

The objective of this study was to determine the biomechanical and microbiological effects of exposing natural hog casings to ozonated water ≈7mg/l for 0, 2 or 4h at 16°C. A total of 450 casing segments representing 10 hanks were used over five testing days and arranged in a randomized block split-plot design. For each treatment, pH, temperature, actual ozone concentration, bursting strength, maximum rupture force, and L(∗), a(∗) and b(∗) color space values were determined. The bursting strength and the maximum rupture force values suggested that casings can be treated by ozone up to 2h without deterioration. After ozone treatments, changes in L(∗), a(∗) and b(∗) color space values made the casings appear lighter than the control samples. Microbiological studies showed that 1 and 2h ozonation reduced counts of Escherichia coli biotype I, which expressed green fluorescent protein, by 0.4 and 0.6log(10)CFU/25.4cm casing, respectively.

Beef hide antimicrobial interventions as a means of reducing bacterial contamination.

This project was designed to evaluate interventions capable of reducing bacterial counts on the hide prior to opening. In Trial I, fresh beef hides (n=12) were cut into sections and assigned to serve as either clipped (hair trimmed) or non-clipped sections. Sections were inoculated with a bovine fecal slurry and sampled following a water wash. Treatments (distilled water, isopropyl alcohol, 3% hydrogen peroxide, 2% l-lactic acid, 10% povidone-iodine, and 1% cetylpyridinium chloride (CPC)) were then applied to each section and the sections were sampled for enumeration of aerobic plate counts (APCs), coliforms, and Escherichia coli. Within clipped samples, 1% CPC and 3% hydrogen peroxide caused the greatest reductions in APCs (4.6 and 4.4 log(10)CFU/100-cm(2), respectively), and 1% CPC, 2% l-lactic acid, and 3% hydrogen peroxide caused the greatest reductions in coliform counts (4.5, 4.1, and 3.9 log(10)CFU/100-cm(2), respectively). In Trial II, beef carcasses with hides on were sampled initially and clipped, and then 2% l-lactic acid, 3% hydrogen peroxide, or 1% CPC were applied before sampling. For APCs, 1% CPC produced the greatest reduction on the hide surface (3.8 log(10)CFU/100-cm(2)). Selective application of these antimicrobials to clipped hide opening sites reduced bacterial counts on hide surfaces, and therefore could reduce final carcass counts in these areas by decreasing the bacterial load before opening.

Identification of Escherichia coli O157:H7 meat processing indicators for fresh meat through comparison of the effects of selected antimicrobial interventions.

Fresh meat products can become contaminated with the pathogen Escherichia coli O157:H7 during the slaughter process; therefore, an E. coli O157:H7 indicator to verify the effectiveness of process controls in slaughter establishments would be extremely useful. The hides of 20 beef cattle were sampled, and 113 bacterial isolates were obtained. Thirteen of these isolates representing four genera, Escherichia, Enterobacter, Providencia, and Serratia, were selected based on growth and biochemical characteristics similar to those of five clinical strains of E. coli O157:H7. The temperature sensitivity was determined for the individual isolates and the five E. coli O157:H7 strains at 55 and 65 degrees C. D65-values for all 13 isolates were not significantly different from D65-values of the E. coli O157:H7 strains. E. coli isolates were the only isolates whose D55-values were not significantly different from those of the E. coli O157:H7 strains. E. coli isolates P3 and P68 were more resistant to the effects of 55 degrees C than were the other E. coli isolates but were not significantly different from E. coli O157:H7 WS 3331 (P > 0.05). The remaining E. coli isolates (P1, P8, and P14) were not significantly different from E. coli O157:H7 strains ATCC 35150, ATCC 43894, ATCC 43895, and WS 3062 (P > 0.05). Prerigor lean and adipose beef carcass tissue was artificially contaminated with stationary-phase cultures of the five E. coli beef cattle isolates or a cocktail of five E. coli O157:H7 strains in a fecal inoculum. Each tissue sample was processed with the following microbial interventions: 90 degrees C water; 90 degrees C water followed by 55 degrees C 2% lactic acid; 90 degrees C water followed by 20 degrees C 2% lactic acid; 20 degrees C water followed by 20 degrees C 2% lactic acid; 20 degrees C water followed by 20 degrees C 20 ppm chlorine; and 20 degrees C water followed by 20 degrees C 10% trisodium phosphate. The appropriateness of the E. coli isolates as potential E. coli O157:H7 indicators was dependent upon the microbial intervention utilized. For all microbial intervention methods applied irrespective of tissue type, the mean log reductions of at least two E. coli isolates were not significantly different from the mean log reduction of the E. coli O157:H7 cocktail (P > 0.05). Because of the frequent employment of multiple microbial interventions in the cattle industry, no single isolate can realistically represent the effectiveness of all microbial interventions for reduction of E. coil O157:H7. Thus, the use of a combination of E. coli isolates may be required to accurately predict the effectiveness of microbial intervention methods on the reduction of E. coli O157:H7 in beef carcass tissue.

Survival of salmonella transformed to express green fluorescent protein on Italian parsley as affected by processing and storage.

To study the effect of processing and storage parameters on the survival of Salmonella on fresh Italian parsley, parsley bunches were dipped for 3 or 15 min in suspensions that were preequilibrated to 5, 25, or 35 degrees C and inoculated with Salmonella transformed to express enhanced green fluorescent protein. Loosely attached and/or associated, strongly attached and/or associated, and internalized and/or entrapped Salmonella cells were enumerated over 0, 1, and 7 days of storage at 25 degrees C and over 0, 1, 7, 14, and 30 days of storage at 4 degrees C using surface-plating procedures. Leaf sections obtained from samples after 0, 1, and 7 days of storage were examined using confocal scanning laser microscopy. Temperature of the dip suspension had little effect on the attachment and survival of Salmonella cells on parsley. Regardless of the temperature or duration of dip, Salmonella was internalized. Immersion for longer times resulted in higher numbers of attached and internalized cells. Microscopic observations supported these results and revealed Salmonella cells near the stomata and within cracks in the cuticle. Storage temperature had the greatest impact on the survival of Salmonella cells on parsley. When stored at 25 degrees C, parsley had a shelf life of 7 days, and Salmonella populations significantly increased over the 7 days of storage. For parsley stored at 4 degrees C, numbers of Salmonella cells decreased over days 0, 1, and 7. After 7 days of storage, there were no viable internalized Salmonella cells detected. Storage temperature represents an important control point for the safety of fresh parsley.

Concentrations of Escherichia coli and genetic diversity and antibiotic resistance profiling of Salmonella isolated from irrigation water, packing shed equipment, and fresh produce in Texas.

Fresh produce has been repeatedly implicated as a vehicle in the transmission of foodborne gastroenteritis. In an effort to assess the risk factors involved in the contamination of fresh produce with pathogenic bacteria, a total of 1,257 samples were collected from cantaloupe, oranges, and parsley (both in the field and after processing) and from the environment (i.e., irrigation water, soil, equipment, etc.). Samples were collected twice per season from two production farms per commodity and analyzed for the presence of Salmonella and Escherichia coli. E. coli was detected on all types of commodities (cantaloupe, oranges, and parsley), in irrigation water, and on equipment surfaces. A total of 25 Salmonella isolates were found: 16 from irrigation water, 6 from packing shed equipment, and 3 from washed cantaloupes. Salmonella was not detected on oranges or parsley. Serotyping, pulsed-field gel electrophoresis (PFGE), and repetitive element sequence-based PCR (rep-PCR) assays were applied to all Salmonella isolates to evaluate the genetic diversity of the isolates and to determine relationships between sources of contamination. Using PFGE, Salmonella isolates obtained from irrigation water and equipment were determined to be different from cantaloupe isolates; however, DNA fingerprinting did not conclusively define relationships between contamination sources. All Salmonella isolates were subjected to antimicrobial susceptibility testing using the disk diffusion method, and 20% (5 of 25) of the isolates had intermediate sensitivity to streptomycin. One Salmonella isolate from cantaloupe was resistant to streptomycin.

Evaluation of peroxyacetic acid as a post-chilling intervention for control of Escherichia coli O157:H7 and Salmonella Typhimurium on beef carcass surfaces.

Four experiments were conducted to test the efficacy of peroxyacetic acid as a microbial intervention on beef carcass surfaces. In these experiments, beef carcass surfaces were inoculated with fecal material (no pathogens) or fecal material containing rifampicin-resistant Escherichia coli O157:H7 and Salmonella Typhimurium. Inoculated surfaces were subjected to a simulated carcass wash with and without 2% l-lactic acid treatment before chilling. In Experiments 1 and 2, the chilled carcass surfaces were sprayed with peroxyacetic acid (200 ppm; 43°) for 15 s. Peroxyacetic acid had no effect on microbial counts of any organism measured on these carcass surfaces. However, lactic acid reduced counts of E. coli Type I (1.9log(10) CFU/cm(2)), coliforms (3.0log(10) CFU/cm(2)), E. coli O157:H7 (2.7log(10) CFU/cm(2)), and S. Typhimurium (2.8log(10) CFU/cm(2)) entering the chilling cooler and prevented growth during the chilling period. In Experiment 3, peroxyacetic acid at different concentrations (200, 600, and 1000 ppm) and application temperatures (45 and 55 °C) were used to investigate its effectiveness in killing E. coli O157:H7 and S. Typhimurium compared to 4% l-lactic acid (55 °C). Application temperature did not affect the counts of either microorganism. Peroxyacetic acid concentrations up to 600 ppm had no effect on these microorganisms. Concentrations of 1000 ppm reduced E. coli O157:H7 and S. Typhimurium by up to 1.7 and 1.3log(10) CFU/cm(2), respectively. However, 4% lactic acid reduced these organisms by 2.7 and 3.4log(10) CFU/cm(2), respectively. In Experiment 4, peroxyacetic acid (200 ppm; 43 °C) was applied to hot carcass surfaces. This treatment caused a 0.7log(10) CFU/cm(2) reduction in both E. coli O157:H7 and S. Typhimurium. The collective results from these experiments indicate that peroxyacetic acid was not an effective intervention when applied to chilled inoculated carcass piece surfaces.

Evaluation of peroxyacetic acid as a potential pre-grinding treatment for control of Escherichia coli O157:H7 and Salmonella Typhimurium on beef trimmings.

Peroxyacetic acid was evaluated in four separate trials for ability to reduce populations of Escherichia coli O157:H7 and Salmonella serotype Typhimurium on fresh beef trim. Trial 1 examined the effectiveness of peroxyacetic acid on individual pieces of fresh beef trim. Trial 2 evaluated the efficacy of peroxyacetic acid at low levels of contamination on batches of fresh beef trim. Trial 3 studied a washing effect of water. Lastly, Trial 4 compared the effectiveness of peroxyacetic acid to lactic acid. At various inoculation levels, peroxyacetic acid reduced populations of both pathogens by approximately 1.0log(10)CFU/cm(2) on fresh beef trim. Trial 3 showed that approximately half of the reductions found in Trials 1 and 2 were due to a washing effect of the water dip. In addition, as shown in Trial 1, increases in concentrations (>200ppm) did not significantly increase log(10) reductions of both pathogens. Following a water dip in Trial 4, peroxyacetic acid caused a reduction of 0.7log(10)CFU/cm(2) in E. coli O157:H7 and 1.0log(10)CFU/cm(2) in Salmonella Typhimurium, whereas lactic acid caused a reduction of 1.3log(10)CFU/cm(2) in E. coli O157:H7 and 2.1log(10)CFU/cm(2) in S. Typhimurium following the water dip. These results show that peroxyacetic acid was not more effective than 2% l-lactic acid in reducing pathogens on fresh beef trim.

Salmonella contamination during production of cantaloupe: a binational study.

Six cantaloupe farms and packing plants in South Texas (950 cantaloupe, 140 water, and 45 environmental samples), including the Rio Grande Valley area, and three farms in Colima State, Mexico (300 cantaloupe, 45 water, and 15 environmental samples), were sampled to evaluate cantaloupe contamination with Salmonella and Escherichia coli during production and processing. Samples collected from external surfaces of cantaloupes, water, and the environments of packing sheds on cantaloupe farms were examined for the presence of Salmonella and E. coli. Of a total of 1,735 samples collected, 31 (1.8%) tested positive for Salmonella. Fifteen Salmonella serotypes were isolated from samples collected in Texas, and nine from samples collected in Colima. Two serotypes (Poona and Oranienburg) that have been associated with three large Salmonella outbreaks in the United States and Canada linked to the consumption of contaminated cantaloupe were found in water samples collected at four farms (three from the United States). Susceptibility of Salmonella isolates to 10 antimicrobials was evaluated by disk diffusion. Eighty-eight percent of the isolates from the United States and Mexico were pansusceptible to the antimicrobials tested; eight isolates from the United States demonstrated an intermediate susceptibility to streptomycin and only two isolates were resistant to the same antimicrobial. From Mexico, four isolates showed an intermediate susceptibility to streptomycin and one isolate was resistant to nalidixic acid and streptomycin. Repetitive sequence-based PCR analysis of Salmonella isolates helped to trace potential sources of Salmonella contamination in source water and in subsequent water samples obtained after the filtration systems of U.S. and Mexican cantaloupe farms. No differences could be seen between the levels of Salmonella contamination in melons from both countries.

Decontamination of cattle hides prior to slaughter using washes with and without antimicrobial agents.

Two trials were conducted to determine the efficacy of cattle wash treatments in reducing pathogens on hides of cattle before slaughter. In trial I, live cattle (n = 120) were washed in an automated, commercial cattle wash system with one of four treatments (single water wash, double water wash, water wash with 0.5% L-lactic acid, or water wash with 50 ppm chlorine). Samples were collected at three locations (brisket, belly, and inside round) pre- and posttreatment to evaluate the effectiveness of treatments on the reduction of aerobic plate counts, coliforms, Escherichia coli and the incidence of Salmonella. For all three locations, bacterial numbers increased from 0.1 to 0.8 log CFU/cm2 posttreatment. In trial II, hide samples were inoculated in the laboratory with 6.0 log CFU/cm2 of rifampicin-resistant Salmonella serotype Typhimurium. Hide wash treatments included higher concentrations of chlorine (100, 200, and 400 ppm) and L-lactic acid (2, 4, and 6%), as well as other antimicrobial agents such as ethanol (70, 80, and 90%), acetic acid (2, 4, and 6%), and Oxy-Sept 333 (0.5, 2, and 4%). Spray wash treatments with ethanol and 4 to 6% concentrations of lactic acid had greater (P < 0.05) mean log reductions than 2% solutions of acetic or lactic acid, as well as 100, 200, and 400 ppm chlorine and the control water wash treatment. Spray wash treatments with Oxy-Sept 333 and 100, 200, or 400 ppm chlorine were not effective (P > 0.05) in reducing Salmonella Typhimurium compared to the (control) distilled water spray wash treatment. Several effective cattle hide interventions were identified in a controlled laboratory setting, but the high concentrations required for effectiveness would likely present problems from an animal welfare standpoint.

Hot water and organic acid interventions to control microbiological contamination on hog carcasses during processing.

Pork skin and muscle tissue were washed with water at temperatures from 25 to 80 degrees C. Water temperatures of 65 and 80 degrees C resulted in greater population reductions of Enterobacteriaceae on pork muscle tissue than lower water temperatures. There was no observable effect of water temperature on population reductions of Enterobacteriaceae on pork skin. Water temperatures of 55, 65, and 80 degrees C reduced the populations of Enterobacteriaceae on inoculated scalded carcasses processed in a university abattoir by 1 to 1.5 log/cm2. Following the water wash with an organic acid rinse resulted in further numerical reductions in populations, although these were not statistically different from the water wash alone. The jowls of both scalded and skinned carcasses processed in a commercial establishment were directly inoculated with a fecal material slurry and then processed with organic acid rinsing only, hot water washing only, or a combination of hot water washing followed by organic acid rinsing. The hot water and acid treatment reduced the populations of mesophilic aerobic bacteria and Escherichia coli by approximately 2 log cycles on both scalded and skinned hog carcasses. The combined treatment resulted in 60% of the scalded carcasses and 40% of the skinned carcasses with undetectable levels of E. coli after direct fecal inoculation of the carcasses. Hot water washing followed by organic acid rinsing can significantly improve the microbiological quality of pork carcasses.

Lactic acid and trisodium phosphate treatment of lamb breast to reduce bacterial contamination.

Lactic acid and trisodium phosphate (TSP) were evaluated for the ability to reduce Escherichia coli and aerobic plate counts (APCs) on lamb breasts that were inoculated with a lamb fecal paste. A 90-s water rinse was applied followed by either a 9-s (55 degrees C) 2% lactic acid spray, a 60-s (55 degrees C) 12% TSP dip, or a combined treatment of both lactic acid and TSP treatments. Lactic acid reduced E. coli and APCs by 1.6 log10/cm2, and TSP caused a 1.8-log10/cm2 reduction in E. coli and a 0.7-log10/cm2 reduction in APCs. Combined reductions by the lactic acid spray followed by the TSP dip were 1.8 and 1.5 log10/cm2 for E. coli and APCs, respectively. Lactic acid and trisodium phosphate, used alone or in combination, were effective in reducing numbers of E. coli and could be useful as pathogen intervention steps in lamb slaughter processing.

In-Plant evaluation of a lactic acid treatment for reduction of bacteria on chilled beef carcasses..

The effectiveness of a lactic acid treatment consisting of spraying a 4% L-lactic acid solution (55 degrees C at source) on chilled beef carcasses to reduce bacterial populations was tested in a commercial slaughter environment. All carcasses had been treated with a proprietary decontamination treatment composed of a hot water spray followed by a lactic acid spray prior to chilling. Bacterial groups used to indicate reductions included aerobic plate count (APC), total coliform count, and Escherichia coli count, and samples were examined from the brisket, the clod, and the neck regions of 40 untreated and 40 treated carcass sides. Depending on the carcass surface region, APCs were reduced by 3.0 to 3.3 log cycles. Log coliform and E. coli counts were consistently reduced to undetectable levels. The small reductions observed for coliforms are attributable to counts on untreated carcasses already being near the lower detection limit of the counting method. The percentage of samples with detectable numbers of coliforms (positive samples) on untreated carcasses ranged from 52.5 to 92.5%, while 0.0% of the samples collected from treated carcasses contained detectable coliforms. Percent E. coli-positive samples ranged from 7.5 to 30.0% on untreated carcasses and 0.0% after treatment of carcass sides. These results indicate that a hot lactic acid spray with increased concentration and time of application may be effectively implemented for an additional decontamination treatment of chilled beef carcasses prior to fabrication.

Lactic acid sprays reduce bacterial pathogens on cold beef carcass surfaces and in subsequently produced ground beef.

Organic acids have been shown to be effective in reducing the presence of pathogenic bacteria on hot beef carcass surfaces; however, application for decontaminating chilled carcasses has not been fully evaluated. In this study, a postchill, 30-s lactic acid spray (500 ml of 4% L-lactic acid, 55 degrees C) was applied onto outside rounds that had been contaminated with Escherichia coli O157:H7 and Salmonella Typhimurium, subsequent to prechill hot carcass treatments consisting of water wash alone or water wash followed by a 15-s lactic acid spray (250 ml of 2% L-lactic acid, 55 degrees C). The prechill treatments reduced both pathogens by 3.3 to 3.4 log cycles (water wash alone) to 5.2 log cycles (water wash and lactic acid). In all cases, the postchill acid treatment produced an additional reduction in E. coli O157:H7 of 2.0 to 2.4 log cycles and of 1.6 to 1.9 log cycles for Salmonella Typhimurium. The counts of both pathogens remained significantly lower in ground beef produced from the outside rounds that received prechill and postchill acid spray than from those that received a postchill spray only. These data indicate that organic acid sprays may be successfully applied for pathogen reduction in beef carcass processing after the cooler, especially when combined with prechill treatments.

Growth of Helicobacter pylori in various liquid and plating media.

The objectives of this research were to compare commonly used liquid and plating media to elucidate whether one medium provided superior growth of Helicobacter pylori in vitro. The liquid media compared were Mueller-Hinton broth, brain heart infusion broth and H. pylori special peptone broth, formulated in this laboratory. No significant differences in growth rates were noted and shaking during the incubation of broths was not essential for good growth. The plating media compared included Columbia agar, Mueller-Hinton agar, modified Glupczynski's Brussels campylobacter charcoal agar, Johnson-Murano agar and H. pylori special peptone agar (HPSPA). None of the non-specific plating media that have been used historically to culture H. pylori exhibited any particular advantage. However, HPSPA provided an obvious advantage in colony size. Helicobacter pylori special peptone agar enhances the cultivation of H. pylori and could improve the recovery of the bacterium from clinical samples in vitro.

Attempts to isolate Helicobacter from cattle and survival of Helicobacter pylori in beef products.

This study focused on important factors related to the potential of cattle and beef products to transmit Helicobacter pylori to humans. Mucosal samples were collected from the rumen and abomasum of 105 cattle and were plated on a selective medium to isolate Helicobacter spp.; none of the samples examined contained these bacteria. Studies were also conducted to determine how long H. pylori survives in refrigerated or frozen ground beef; results indicated that the microorganism dies rapidly in ground beef, whether refrigerated or frozen. Packaging in vacuum or air had little effect on survival of the organism. The number of H. pylori decreased in refrigerated samples from 3.3 log10 CFU/g on day 0 to 1.4 log10 CFU/g on day 6. H. pylori died even more rapidly when frozen, decreasing from 3.3 log10 CFU/g on day 0 to 0.5 log10 CFU/g on day 6. Retail beef cuts (n = 20) were also examined for the presence of H. pylori by direct plating on a selective medium and by incubation in an enriched broth followed by plating on a selective medium. None of the retail samples contained H. pylori. This research suggests that transmission of H. pylori from beef and beef products is not a primary factor in the high prevalence of this bacterium in humans.

Development of a selective medium for isolation of Helicobacter pylori from cattle and beef samples.

Helicobacter pylori has been isolated from the human stomach with media containing only minimal selective agents. However, current research on the transmission and sources of infection requires more selective media due to the higher numbers of contaminants in environmental, oral, and fecal samples. The objective of this study was to develop and evaluate detection techniques that are sufficiently selective to isolate H. pylori from potential animal and food sources. Since H. pylori survives in the acidic environment of the stomach, low pH with added urea was studied as a potential selective combination. H. pylori grew fairly well on H. pylori Special Peptone plating medium supplemented with 10 mM urea at pH 4. 5, but this pH did not sufficiently inhibit the growth of contaminants. Various antibiotic combinations were then compared, and a combination consisting of 10 mg of vancomycin per liter, 5 mg of amphotericin B per liter, 10 mg of cefsulodin per liter, 62,000 IU of polymyxin B sulfate per liter, 40 mg of trimethoprim per liter, and 20 mg of sulfamethoxazole per liter proved to be highly selective but still allowed robust colonies of H. pylori to grow. This medium was highly selective for recovering H. pylori from cattle and beef samples, and it is possible that it could be used to enhance the recovery of this bacterium from human and environmental samples, which may be contaminated with large numbers of competing microorganisms.

Reduction of Escherichia coli O157:H7 and Salmonella typhimurium on beef carcass surfaces using acidified sodium chlorite.

The efficacy of a phosphoric acid-activated acidified sodium chloride (PASC) spray and a citric acid-activated acidified sodium chlorite (CASC) spray applied at room temperature (22.4 to 24.7 degrees C) in combination with a water wash was compared with that of a water wash only treatment for reduction of Escherichia coli O157:H7 and Salmonella Typhimurium inoculated onto various hot-boned individual beef carcass surface regions (inside round, outside round, brisket, flank, and clod). Initial counts of 5.5 and 5.4 log CFU/cm2 were obtained after inoculation with E. coli O157:H7 and Salmonella Typhimurium, respectively. Initial numbers for both pathogens were reduced by 3.8 to 3.9 log cycles by water wash followed by PASC spray and by 4.5 to 4.6 log cycles by water wash followed by CASC spray. The sprays consisted of applying 140 ml of the appropriate sanitizing solution for 10 s at 69 kPa. Corresponding reduction values obtained by water wash alone were 2.3 log. The performance of CASC appeared to be consistently better than that of PASC. In general, no effect of the carcass surface region was observed on the log reductions for either pathogen, except for the inside round, which consistently had lower reductions. Both PASC and CASC were capable of effectively reducing pathogens spread to areas beyond the initial contaminated area of the cuts to levels close to or below the counting method detection limit (0.5 log CFU/cm2). However, 30 to 50% of the carcasses treated by these antimicrobial solutions still yielded countable colonies. Results of this study indicate that acidified sodium chlorite sprays are effective for decontaminating beef carcass surfaces.