Mindy M. Brashears

Prevalence of Escherichia coli O157:H7 and performance by beef feedlot cattle given Lactobacillus direct-fed microbials.

Fecal shedding of Escherichia coli O157:H7, the prevalence of Escherichia coli O157:H7 in pens and on carcasses and hides, and cattle performance as a result of daily dietary supplementation with Lactobacillus-based direct-fed microbials (DFMs) were evaluated in a feeding trial involving 180 beef steers. Steers were evaluated for shedding of E. coli O157:H7 by an immunomagnetic separation technique on arrival at the feedlot, just before treatment with the DFMs, and every 14 days thereafter until slaughter. Composite pen fecal samples were collected every 14 days (alternating weeks with animal testing), and prevalence on hides and carcasses at slaughter was also evaluated. Feedlot performance (body weight gain and feed intake) was measured for the period during which the DFMs were fed. Gain efficiency was calculated as the ratio of weight gain to feed intake. Lactobacillus acidophilus NPC 747 decreased (P < 0.01) the shedding of E. coli O157:H7 in the feces of individual cattle during the feeding period. E. coli O157:H7 was approximately twice as likely to be detected in control animal samples as in samples from animals receiving L. acidophilus NPC 747. In addition, DFM supplementation decreased (P < 0.05) the number of E. coli O157:H7-positive hide samples at harvest and the number of pens testing positive for the pathogen. Body weight gains (on a live or carcass basis) and feed intakes during the DFM supplementation period did not differ among treatments. Gain efficiencies on a live-weight basis did not differ among treatments, but carcass-based gain/feed ratios tended (P < 0.06) to be better for animals receiving the two DFM treatments than for control animals. The results of this study suggest that the feeding of a Lactobacillus-based DFM to cattle will decrease, but not eliminate, fecal shedding of E. coli O157:H7, as well as contamination on hides, without detrimental effects on performance.

Pre-harvest interventions to reduce carriage of E. coli O157 by harvest-ready feedlot cattle.

Escherichia coli O157 is an important cause of food-borne illness. The primary reservoir for this organism is cattle and at present the major site of control is within abattoirs. Recent data have highlighted the importance of the pathogen load entering abattoirs on harvest-ready feedlot cattle. The likelihood for in-plant intervention failure increases as the proportion of cattle carrying E. coli O157 within a pen increases. Pre-harvest reduction of E. coli O157 colonization will require targeted intervention strategies and should reduce contamination of carcasses thereby enhancing public health. Several pre-harvest interventions show substantial promise, such as specific strains of direct-fed microbials, vaccine technology, sodium chlorate, and neomycin sulfate, whereas others such as Brown Seaweed or chlorination of water have little or no detectable benefit. Selection of validated interventions strategies will be important as efforts to control pre-harvest carriage of E. coli O157 increase.

Reduction of Escherichia coli O157 in finishing beef cattle by various doses of Lactobacillus acidophilus in direct-fed microbials.

Our objective was to evaluate the effects of three doses of Lactobacillus acidophilus strain NP51 and a combination treatment of strains NP51 and NP45 on prevalence of Escherichia coli O157 in cattle. Three hundred steers were assigned randomly to 60 pens (five steers per pen) and received one of five treatments: (i) control, no added direct-fed microbial; (ii) HNP51, high dose of NP51 at 10(9) CFU per steer daily; (iii) MNP51, NP51 at 10(8) CFU per steer daily; (iv) LNP51, low dose of NP51 at 10(7) CFU per steer daily; and (v) NP51+45, NP51 at 10(9) CFU per steer daily and NP45 at 106 CFU per steer daily. All direct-fed microbial treatments included Propionibacterium freudenreichii at 10(9) CFU per steer. Individual rectal fecal samples were collected on arrival and every 28 days throughout the feeding period. Fecal and hide samples were collected on the day of harvest. Samples were analyzed for presence of E. coli O157 using immunomagnetic separation methods. Cattle receiving HNP51, MNP51, and LNP51 had a lower prevalence (P < 0.01) of E. coli O157 throughout the feeding period compared with the controls, and the dose response for NP51 was a linear decrease in prevalence with increasing dose (P < 0.01). No decrease in prevalence for cattle receiving the combination NP51+45 was detected compared with controls (P = 0.15). E. coli O157 prevalences averaged across collection times were 23.9, 10.5, 9.9, 6.8, and 17.3% for cattle in the control, LNP51, MNP51, HNP51, and NP51 +45 groups, respectively. Least squares mean estimates of fecal prevalence at harvest of E. coli O157 were 31.7, 12.5, 17.4, 8.2, and 41.6% among cattle in the control, LNP51, MNP51, HNP51, and NP51+45 groups, respectively. Least squares mean estimates of the percentage of positive hide samples at harvest were 8.7, 5.9, 4.8, 3.4, and 8.6% among cattle in the control, LNP51, MNP51, HNP51, and NP51+45 groups, respectively. The greatest decrease in E. coli O157 carriage was achieved using NP51 at 10(9) CFU per steer.

Dietary Supplementation with Lactobacillus- and Propionibacterium-Based Direct-Fed Microbials and Prevalence of Escherichia coli O157 in Beef Feedlot Cattle and on Hides at Harvest

The objective of this study was to describe the prevalence of Escherichia coli O157 in the feces and on the hides of finishing beef cattle fed a standard diet and those fed diets supplemented with direct-fed microbials. Two hundred forty steers received one of four treatments throughout the feeding period: (i) control: no added microbials; (ii) HNP51: high dose of Lactohacillius acidophilus strain NP 51 (10(9) CFU per steer daily) and Propionibacterium freudenreichii (10(9) CFU per steer daily); (iii) HNP51+45: high dose of NP 51 (10(9) CFU per steer daily), P. freudenreichii (10(9) CFU per steer daily), and L. acidophilus NP 45 (10(6) CFU per steer daily); or (iv) LNP51+45: low dose of NP 51 (10(6) CFU per steer daily), P. freudenreichii (10(9) CFU per steer daily), and NP 45 (10(6) CFU per steer daily). Samples were collected from each animal and analyzed for the presence of E. coli O157 using immunomagnetic separation methods on day 0 (feces), 7 days before harvest (feces), and at harvest (feces and hide). At the end of the feeding period, cattle receiving HNP51 were 57% less likely to shed detectable E. coli O157 in their feces than were the controls (P < 0.01). For animals receiving HNP51+45 and LNP51+45, fecal prevalence did not differ from that of the controls. The prevalence of positive hide samples was least among cattle receiving HNP51+45 (3.3%); these animals were 79% less likely (P < 0.06) to have a positive hide sample than were the controls (prevalence = 13.8%). There was poor agreement of the culture results between fecal and hide samples collected from the same animal (kappa = 0.08; confidence interval = -0.05 to 0.2). Cattle supplemented with a high dose of NP 51 had reduced E. coli O157 prevalence in both fecal and hide samples, indicating that this treatment may be an efficacious preharvest intervention strategy.

Reduction of Escherichia coli O157 and Salmonella in Feces and on Hides of Feedlot Cattle Using Various Doses of a Direct-Fed Microbial

In this study, the effectiveness of direct-fed microbials at reducing Escherichia coli O157 and Salmonella in beef cattle was evaluated. Steers (n=240) received one of the following four treatment concentrations: control = lactose carrier only; low = 1 X 10(7) CFU per steer daily Lactobacillus acidophilus NP51; medium = 5 x 10(8) CFU per steer daily L. acidophilus NP51; and high = 1 x 10(9) CFU per steer daily L. acidophilus NP51. Low, medium, and high diets also included 1 x 10(9) CFU per steer Propionibacterium freudenreichii NP24. Feces were collected from each animal at allocation of treatment and found to have no variation (P = 0.54) between cohorts concerning E. coli O157 recovery. Feces and hide swabs were collected at harvest and analyzed for the presence of E. coli O157 by immunomagnetic separation and Salmonella by PCR. No significant dosing effects were detected for E. coli O157 recovery from feces at the medium dose or from hides at the medium and high doses. E. coli O157 was 74% (P < 0.01) and 69% (P < 0.01) less likely to be recovered in feces from animals receiving the high and low diets, respectively, compared with controls. Compared with controls, E. coli O157 was 74% (P = 0.05) less likely to be isolated on hides of cattle receiving the low dose. No significant dosing effects were detected for Salmonella recovery from feces at the medium and low doses or from hides at any doses. Compared with controls, Salmonella was 48% (P = 0.09) less likely to be shed in feces of cattle receiving the high dose. No obvious dose-response of L. acidophilus NP51 on recovery of E. coli O157 or Salmonella was detected in our study.

Microbial profile and antibiotic susceptibility of Campylobacter spp. and Salmonella spp. in broilers processed in air-chilled and immersion-chilled environments.

Carcass chilling is considered a critical step for inhibiting bacterial growth during poultry processing. The objective of this study was to compare microbiological loads and the incidence of Salmonella spp. and Campylobacter spp. on broiler carcasses subjected to immersion chilling and air chilling. Additionally, the antibiotic resistance patterns of pathogen isolates were determined. The results of this study indicated that the incidence of Salmonella spp. and Campylobacter spp. tends to be significantly lower in air-chilled broilers, suggesting that cross-contamination may be more prevalent for immersion-chilled broilers. No significant differences were detected between chilling treatments for total aerobic populations or for generic E. coli or coliform counts. Psychrotrophic populations were significantly larger (P < 0.05) in immersion-chilled broilers than in their air-chilled counterparts. Campylobacter isolates from immersion-chilled broilers had a higher incidence of resistance to nalidixic acid (NAL) and related fluoroquinolones than isolates from air-chilled broilers did. Additionally, Campylobacter isolates from air-chilled broilers had a higher frequency of resistance to tetracycline than isolates from immersion-chilled broilers did. With regard to Salmonella, isolates from immersion-chilled broilers had a higher incidence of resistance to NAL than isolates from air-chilled samples did. No Salmonella isolates from immersion- or air-chilled broilers were resistant to the fluoroquinolones tested. The chilling method used during processing may influence the microbial profile of postchilled broilers.

Validation of the use of organic acids and acidified sodium chlorite to reduce Escherichia coli O157 and Salmonella typhimurium in beef trim and ground beef in a simulated processing environment

A study was conducted to determine if acidified sodium chlorite (1,200 ppm) and acetic and lactic acids (2 and 4%) were effective in reducing foodborne pathogens in beef trim prior to grinding in a simulated processing environment. The reduction of Salmonella Typhimurium and Escherichia coli O157:H7 at high (4.0 log CFU/g) and low (1.0 log CFU/g) inoculation doses was evaluated at various processing steps, including the following: (i) in trim just after treatment application, (ii) in ground beef just after grinding, (iii) in ground beef 24 h after refrigerated storage, (iv) in ground beef 5 days after refrigerated storage, and (v) in ground beef 30 days after frozen storage. All antimicrobial treatments reduced the pathogens on the trim inoculated with the lower inoculation dose to nondetectable numbers in the trim and in the ground beef. There were significant reductions of both pathogens in the trim and in the ground beef inoculated with the high inoculation doses. On the trim itself, E. coli O157:H7 and Salmonella Typhimurium were reduced by 1.5 to 2.0 log cycles, with no differences among all treatments. In the ground beef, the organic acids were more effective in reducing both pathogens than the acidified sodium chlorite immediately after grinding, but after 1 day of storage, there were no differences among treatments. Overall, in the ground beef, there was a 2.5-log reduction of E. coli O157:H7 and a 1.5-log reduction of Salmonella Typhimurium that was sustained over time in refrigerated and frozen storage. Very few sensory differences between the control samples and the treated samples were detected by a consumer panel. Thus, antimicrobial treatments did not cause serious adverse sensory changes. Use of these antimicrobial treatments can be a promising intervention available to ground beef processors who currently have few interventions in their process.

Antagonistic action of cells of Lactobacillus lactis toward Escherichia coli O157:H7 on refrigerated raw chicken meat.

Cells of a strain of Lactobacillus lactis selected for ability to produce hydrogen peroxide were added to Trypticase soy broth (TSB) containing Escherichia coli O157:H7 to determine if L. lactis was antagonistic towards the E. coli during storage at 7 degrees C for 7 days. E. coli was enumerated on violet red bile agar. Three strains of E. coli O157:H7 (43894, 43890, and 35150) were evaluated. Control samples containing no L. lactis did not show significant declines in numbers of E. coli during the 7 days of storage. however, samples inoculated with at least 5.0 x 10(7) L. lactis per ml exhibited significant declines in numbers of E. coli after only 3 days of storage for all strains. Samples inoculated with fewer L. lactis displayed vary effects on E. coli O157:H7 depending on the strain E. coli O157:H7 strain 43894 appeared to be the most resistant to the antagonistic action of the L. lactis. Interaction experiments in the presence of catalase indicated that hydrogen peroxide was the main factor responsible for the inhibitory action produced by the lactobacilli. Raw chicken breast meat inoculated with E. coli O157:H7 strain 43894 plus the cells of L. lactis and stored at 5 degrees C exhibited declines in numbers of the pathogen, whereas those inoculated only with the E. coli exhibited no declines during storage at 5 degrees C.

Reduction of Escherichia coli O157:H7 and Salmonella in Ground Beef Using Lactic Acid Bacteria and the Impact on Sensory Properties

Studies were conducted to determine whether four strains of lactic acid bacteria (LAB) inhibited Escherichia coli O157: H7 and Salmonella in ground beef at 5 degrees C and whether these bacteria had an impact on the sensory properties of the beef. The LAB consisted of frozen concentrated cultures of four Lactobacillus strains, and a cocktail mixture of streptomycin-resistant E. coli O157:H7 and Salmonella were used as pathogens. Individual LAB isolates at 10(7) CFU/ml were added to tryptic soy broth containing a pathogen concentration of 10(5) CFU/ml. Samples were stored at 5 degrees C, and pathogen populations were determined on days 0, 4, 8, and 12. After 4 days of storage, there were significant differences in numbers of both pathogens exposed to LAB isolates NP 35 and NP 3. After 8 and 12 days of storage, all LAB reduced populations of both pathogens by an average of 3 to 5 log cycles. A second study was conducted in vacuum-packaged fresh ground beef. The individual LAB isolates resulted in an average difference of 1.5 log cycles of E. coli O157:H7 after 12 days of storage, and Salmonella populations were reduced by an average of 3 log cycles. Following this study, a mixed concentrated culture was prepared from all four LAB and added to ground beef inoculated with pathogen at 10(8) CFU/g. After 3 days of storage, the mixed culture resulted in a 2.0-log reduction in E. coli O157:H7 compared with the control, whereas after 5 days of storage, a 3-log reduction was noted. Salmonella was reduced to nondetectable levels after day 5. Sensory studies on noninoculated samples that contained LAB indicated that there were no adverse effects of LAB on the sensory properties of the ground beef. This study indicates that adding LAB to raw ground beef stored at refrigeration temperatures may be an important intervention for controlling foodborne pathogens.

Spoilage and safety characteristics of ground beef packaged in traditional and modified atmosphere packages.

Two separate studies, one with pathogen-inoculated product and one with noninoculated product, were conducted to determine the safety and spoilage characteristics of modified atmosphere packaging (MAP) and traditional packaging of ground beef patties. Ground beef patties were allotted to five packaging treatments (i) control (foam tray with film overwrap; traditional), (ii) high-oxygen MAP (80% 02, 20% CO2), (iii) high-oxygen MAP with added rosemary extract, (iv) low-oxygen carbon monoxide MAP (0.4% CO, 30% CO2, 69.6% N2), and (v) low-oxygen carbon monoxide MAP with added rosemary extract. Beef patties were evaluated for changes over time (0, 1, 3, 5, 7, 14, and 21 days) during lighted display. Results indicated low-oxygen carbon monoxide gas flush had a stabilizing effect on meat color after the formation of carboxymyoglobin and was effective for preventing the development of surface discoloration. Consumers indicated that beef patties packaged in atmospheres containing carbon monoxide were more likely to smell fresh at 7, 14, and 21 days of display, but the majority would probably not consume these products after 14 days of display because of their odor. MAP suppressed the growth of psychrophilic aerobic bacteria when compared with control packages. Generally, control packages had significantly higher total aerobic bacteria and Lactobacillus counts than did modified atmosphere packages. In the inoculated ground beef (approximately 10(5) CFU/g) in MAP, Escherichia coli O157 populations ranged from 4.51 to 4.73 log CFU/g with no differences among the various packages, but the total E. coli O157:H7 in the ground beef in the control packages was significantly higher at 5.61 log CFU/g after 21 days of storage. On days 14 and 21, the total Salmonella in the ground beef in control packages was at 5.29 and 5.27 log CFU/g, respectively, which was significantly higher than counts in the modified atmosphere packages (3.99 to 4.31 log CFU/g on day 14 and 3.76 to 4.02 log CFU/g on day 21). Data from these studies indicate that MAP suppresses pathogen growth compared with controls and that spoilage characteristics developed in MAP packages.