Steven E. Niebuhr

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.

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.

The effect of pH and nitrite concentration on the antimicrobial impact of celery juice concentrate compared with conventional sodium nitrite on Listeria monocytogenes

The objectives of this study were to evaluate the impact of pH and nitrite from celery juice concentrate (CJ) on the growth of Listeria monocytogenes in broth and on ham slices, and to evaluate the impact of pH and nitrite from CJ on quality attributes of the ham. The pH of both broth and ham were increased by the addition of CJ. The CJ was less effective than conventional nitrite at 100 mg/kg nitrite in broth, but in ham, the CJ treatments at both 100 and 200 mg/kg resulted in growth of L. monocytogenes (p>0.05) similar to that of the conventional nitrite at the same concentrations. Reducing the pH of CJ before addition to the ham had greater impact on L. monocytogenes growth at 200 mg/kg nitrite than at 100 mg/kg. Celery juice concentrate may increase meat product pH which could have implications for the antimicrobial impact of nitrite in some products.

Evaluation of Escherichia coli Biotype I as a Surrogate for Escherichia coli O157:H7 for Cooking, Fermentation, Freezing, and Refrigerated Storage in Meat Processes

Five Escherichia coli biotype I isolates were compared with E. coli O157:H7 under four common meat processing conditions. The processes that were evaluated were freezing, refrigerating, fermentation, and thermal inactivation. For each study, at least one surrogate organism was not statistically different when compared with E. coli O157:H7. However, the four studies did not consistently show the same isolate as having this agreement. The three studies that involved temperature as a method of controlling or reducing the E. coli population all had at least one possible surrogate in common. In the fermentation study, only one isolate (BAA-1429) showed no statistical difference when compared with E. coli O157:H7. However, the population reductions that were observed indicated the isolates BAA-1427 and BAA-1431 would overestimate the surviving E. coli O157:H7 population in a fermented summer sausage. When all of the data from all of the surrogates were examined, it was found that isolates BAA-1427, BAA-1429, and BAA-1430 would be good surrogates for all four of the processes that were examined in this study. There was no statistical difference noted between these three isolates and E. coli O157:H7 in the refrigeration study. These isolates resulted in smaller population reductions than did E. coli O157:H7 in the frozen, fermentation, and thermal inactivation studies. This would indicate that these isolates would overpredict the E. coli O157:H7 population in these three instances. This overprediction results in an additional margin of safety when using E. coli biotype 1 as a surrogate.

Survival of Listeria monocytogenes and Escherichia coli O157:H7 during Sauerkraut Fermentation

Sauerkraut was produced from shredded cabbage, as is typical in the United States, and from whole head cabbages, which is a traditional process in parts of Eastern Europe. The sauerkraut was inoculated with five strain mixtures of Escherichia coli O157:H7 and Listeria monocytogenes, and the populations of these bacteria, as well as lactic acid bacteria, pH, and titratable acidity, were monitored over the course of fermentation. Fermentation variables were temperature (18 and 22 degrees C) and salt concentration (1.8, 2.25, and 3%). For most of the analyses, the type of cabbage processing was a significant factor, although within cabbage type, neither salt nor fermentation temperature had significant effects. The final pH of the whole-head sauerkraut was lower than the shredded sauerkraut, but the titratable acidity was significantly higher in the shredded sauerkraut. E. coli O157:H7 and L. monocytogenes persisted in the brines for most of the fermentation, although at the end of the fermentations (15 days for shredded, 28 days for whole head), neither pathogen had detectable populations. E. coli populations decreased more rapidly in the shredded sauerkraut even though the pH was higher because of the higher total acidity in the shredded sauerkraut. Acid-tolerant strains of E. coli and L. monocytogenes were isolated from both shredded and whole-head sauerkraut at different salt concentrations and temperatures after 15 days of fermentation and could be detected at 35 days in the wholehead sauerkraut.

Impact of pH Enhancement on Populations of Salmonella, Listeria monocytogenes, and Escherichia coli O157:H7 in Boneless Lean Beef Trimmings †

Boneless lean beef trimmings were inoculated with multiple strains of salmonellae, Listeria monocytogenes, and Escherichia coli O157:H7 at levels of ca. 6 log10 CFU/g. pH enhancement with ammonia gas was then used to increase the pH of the trimmings to ca. 9.6. The product was then frozen, chipped, and compressed into blocks. pH enhancement reduced the populations of salmonellae, L. monocytogenes, and E. coli O157:H7 by approximately 4, 3, and 1 log10 cycles, respectively. After the product had been frozen and compressed into blocks, no salmonellae or E. coli O157:H7 were detectable by enumeration or after enrichment and isolation. The final populations of L. monocytogenes were reduced by ca. 3 log10 cycles relative to the initial populations. When uninoculated pH-enhanced lean boneless trimmings were blended with inoculated ground beef to a final concentration of 15% (wt/wt), pathogen populations in the ground beef were reduced by approximately 0.2 log10 cycles.

Impact of the population of spoilage microflora on the growth of Listeria monocytogenes on frankfurters

Approximately 100 CFU/cm2 of a five-strain mixture of Listeria monocytogenes was coinoculated onto frankfurters with three different concentrations (10(2), 10(4), and 106 CFU/cm2) of an undefined spoilage microflora derived from commercial frankfurters. The frankfurters were vacuum packaged and stored at 10 degrees C for up to 48 days. The populations of L. monocytogenes, aerobic mesophilic bacteria, lactic acid bacteria, and Enterobacteriaceae were determined at various time intervals during storage. After 14 days, the population of L. monocytogenes was highest when grown with a spoilage microflora population of 10(2) CFU/cm2, and this trend continued until 48 days. Throughout the entire storage period, the populations of L. monocytogenes at any concentration of inoculated spoilage microflora rarely differed by more than 0.5 log CFU/cm2, and the maximum observed difference as 1.1 log CFU/cm2 at 40 days. The growth rate of L. monocytogenes was approximately the same at all concentrations of the inoculated spoilage microflora. These results suggest that the concentration of spoilage microflora present on the original processed meat may have a slight impact on the growth of L. monocytogenes in the package.

Transfer of methicillin-resistant Staphylococcus aureus from retail pork products onto food contact surfaces and the potential for consumer exposure.

Methicillin-resistant Staphylococcus aureus (MRSA) is a pathogen that has developed resistance to beta-lactam antibiotics and has been isolated at low population numbers in retail meat products. The objectives of this study were to estimate the potential transfer of MRSA from contaminated retail pork products to food contact surfaces and to estimate the potential for human exposure to MRSA by contact with those contaminated surfaces. Pork loins, bacon, and fresh pork sausage were inoculated with a four-strain mixed MRSA culture over a range of populations from approximately 4 to 8 log, vacuum packaged, and stored for 2 weeks at 5°C to simulate normal packaging and distribution. Primary transfer was determined by placing inoculated products on knife blades, cutting boards, and a human skin model (pork skin) for 5 min. Secondary transfer was determined by placing an inoculated product on the contact surface, removing it, and then placing the secondary contact surface on the initial contact surface. A pork skin model was used to simulate transfer to human skin by placing it into contact with the contact surface. The percentages of transfer for primary transfer from the inoculated products to the cutting board ranged from 39 to 49%, while the percentages of transfer to the knife ranged from 17 to 42%. The percentages of transfer from the inoculated products to the pork skin ranged from 26 to 36%. The secondary transfer percentages ranged from 2.2 to 5.2% across all products and contact surfaces. Statistical analysis showed no significant differences in the amounts of transfer between transfer surfaces and across cell concentrations.

Effect of gamma or beta radiation on Salmonella DT 104 in ground pork.

Mixtures of six Salmonella Typhimurium DT 104 strains were inoculated into three ground pork products to determine the effect of fat content on the radiation resistance of Salmonella DT 104. The ground pork products were 90% lean, 50:50 fat:lean, and 100% fat. Inoculated products were irradiated using a gamma radiation source in a self-contained 137Cesium irradiator or a 10 MeV accelerator producing electrons (e-beam). The radiation D10-values (dose required for a 90% inactivation of viable CFU) for Salmonella DT 104 inoculated into 90% lean ground pork, 50:50 fat/lean ground pork, and 100% pork fat and subjected to beta radiation were 0.42 kGy, 0.43 kGy, and 0.43 kGy, respectively. The corresponding radiation D10-values for Salmonella DT 104 subject to gamma radiation were 0.56, 0.62, and 0.62 kGy, respectively. There was no statistical significant difference (P = 0.3) in radiation D10-values for Salmonella in the three products subject to either radiation treatment. Therefore, fat content had no effect. There was a significant difference (P = 0.001) between the radiation D10-values obtained with the two radiation sources. The radiation D10-values were within the reported range for irradiation destruction of Salmonella contaminated raw meat products.

Can supplemental nitrate in cured meats be used as a means of increasing residual and dietary nitrate and subsequent potential for physiological nitric oxide without affecting product properties

The effects of formulated sodium nitrate plus supplemental nitrate (SN) from celery juice powder on residual nitrite, residual nitrate, rancidity, microbial growth, color, sensory properties, and proximate composition of frankfurters, cotto salami and boneless ham during storage (1°C) were studied. The products were assigned one of two treatments, which were each replicated twice: control (156ppm sodium nitrite) or SN (156ppm sodium nitrite and 1718ppm sodium nitrate in combination with 2% VegStable 502). Sensory parameters and proximate composition were measured once for each replication. All other analytical measurements were conducted at regular intervals for 97-98days. The SN showed no increase in residual nitrite compared to the control. No changes (P>0.05) were observed in residual nitrate during storage for any of the products. The results showed that addition of SN did not significantly alter most physical, chemical or microbial properties of cured meat products during refrigerated storage, but some product dependent sensory effects were observed.