G.R. Schmidt

Evaluation of hand-trimming, various sanitizing agents, and hot water spray-washing as decontamination interventions for beef brisket adipose tissue

Various chemical solutions (5% hydrogen peroxide, 0.5% ozone, 12% trisodium phosphate, 2% acetic acid, and 0.3% commercial sanitizer), water (16 to 74°C) spray-washing interventions, and hand-trimming/spray-washing treatments were compared for their ability to remove fecal material and to reduce bacterial contamination on beef brisket fat samples in a model spray-washing cabinet. The samples were inoculated with 2.5 cm2 of a bovine fecal paste inoculated with Escherichia coli (ATCC 11370). Hand-trimming followed by spray-washing with plain water (16 to 74°C when it came in contact with the sample; 20.68 bar pressure; for 36 or 12 s corresponding to chain speeds of 100 or 300 carcasses per h) lowered (P < 0.05) microbiological counts, compared to the inoculated control, by 1.41 to 2.50 log colony-forming units (CFU)/cm2. Additionally, spraying with chemical solutions (16°C; 1.38 bar, 12 or 36 s), before or after spray-washing with plain water (20.68 bar) of 16°C (36 s), 35°C (12 s) or 74°C (12 s) reduced bacterial counts by 1.34 to 2.87, 1.18 to 2.86, or 0.96 to 3.42 log CFU/cm2, respectively. Reduction in counts was influenced by water temperature (16 to 74°C), type of chemical solution, and sequence of spray application. Under the conditions of this study, hydrogen peroxide and ozonated water were more effective (P < 0.05) than trisodium phosphate, acetic acid, and a commercial sanitizer when applied after first washing with plain water. Trisodium phosphate maintained its activity when used before washing with water. In general, water of 74°C caused reductions (P < 0.05) exceeding 3.0 log CFU/cm2, which were higher than those achieved by trimming and spray-washing. No spreading of bacteria in areas immediately adjacent to the inoculation site was detected following spray-washing.

Growth variation among species and strains of Listeria in culture broth

Culture suspensions of 45 species and strains of Listeria were prepared in tryptic soy broth with 0.6% yeast extract (TSBYE) for 24 h at 37°C, and were then diluted with phosphate buffer solution and standardized to 0.10 ± 0.01 absorbance at 600 nm. Spectrophotometer tubes containing 5 ml of TSBYE (pH 7.2) were inoculated with 0.1 ml of the standardized cultures and incubated at 4, 10 or 37°C. Absorbance readings were taken during storage. Growth curves were fitted using the Gompertz function, and growth parameters were calculated. There were major differences in lag phase duration (LPD), generation time (GT) and exponential growth rate (EGR) among species and strains of Listeria tested. Values for LPD and GT decreased (P <0.05) with increasing temperature of incubation, while EGR and maximum population density (MPD) values increased. Lag phase duration and GT values at a given temperature were lower for Listeria monocytogenes compared to other Listeria spp. At 4°C, LPDs for L. monocytogenes strains ranged from 69.8 to 270.8 h. Of the L. monocytogenes cultures tested, strain Scott A had the longest average (209.8 ± 0.1) h LPD at 4°C. At l0°C, LPDs ranged from 36.5 to 68.9 h, with Scott A being again one of the strains with the longest average LPD (62.8 ± 0.7 h). At 37°C, LPDs ranged from 4.4 to 11.1 h. Variation was also observed in GT and EGR, especially at 4°C. Although there were major variations in growth parameters due to strain and temperature, no significant (P >0.05) trends were observed in average values among different serotypes of L. monocytogenes tested.

An Enzyme-Linked Immunosorbent Assay for Glial Fibrillary Acidic Protein as an Indicator of the Presence of Brain or Spinal Cord in Meat

The current methods to detect central nervous system (CNS) tissue in blood, lungs, or meat are cumbersome, time consuming, and costly. The objective of this study was to use glial fibrillary acidic protein (GFAP), which is restricted to the CNS, in an enzyme-linked immunosorbent assay (ELISA) for the detection of CNS tissue in blood and muscle from beef cattle. Bovine brain, cerebral cortex, spinal cord, sciatic nerve, diaphragm, blood clots, and other skeletal muscle were obtained from three animals at slaughter. The limit for detection of GFAP was approximately 1.0 ng and the standard curve was linear up to 40 ng. Tissue samples gave responses parallel to the GFAP standard, suggesting that standard and unknown samples were immunoreactively identical. No GFAP was detected in skeletal muscle (ground beef, shoulder clod, and diaphragm) and blood clots. Trace amounts (13.5 to 51 ng/mg) were present in sciatic nerve. In contrast, high levels of GFAP (55 to 220 microg/ mg) were present in spinal cord, cerebral cortex (17 microg/mg), and whole brain (9 to 55 microg/mg). In a storage study using two animals in two separate studies, immunoreactive GFAP was detectable for up to 8 days at 4 degrees C in all tissues containing neural elements. Thus, mixtures of muscle with spinal cord or brain retained almost 80% of their immunoreactivity after 8 days at 4 degrees C, while brain and spinal cord alone retained approximately 50% and 25%, respectively, of their initial activities. In a repeat experiment, 80 to 100% of the initial activity was retained in these tissues after 8 days at 4 degrees C. The results of the current study demonstrate that the GFAP ELISA provides a valid and repeatable method to detect CNS tissue contamination in meat.

Effect of meat curing ingredients on thermal destruction of Listeria monocytogenes in ground pork

Ground pork (15% fat) inoculated with a mixture of nine strains of Listeria monocytogenes (107-108 CFU/g) was mixed with different levels of curing ingredients, filled into baby food jars (140 ± 5 g/jar), and cooked in a waterbath to 60°C internal product temperature. Heating rates were similar among meat treatments with different additives. However, destruction of L. monocytogenes was 3 log per gram less in ground pork with 3% sodium chloride than in ground pork without added sodium chloride (P<0.05). The protective effect of sodium chloride against destruction of L. monocytogenes increased with increasing (0-3%) sodium chloride concentration. Dextrose (1%) and a phosphate mixture (0.4%) also protected L. monocytogenes (P<0.05) from thermal destruction. Sodium nitrite (0.0156%) and sodium erythorbate (0.055%) did not affect (P>0.05) the degree of thermal destruction of L. monocytogenes in ground pork. The greatest protective effect was observed when all curing ingredients were combined and added in ground pork. These results indicated that the chance for L. monocytogenes to survive during heat processing is greater in cured than in fresh meat.

The Detection of Central Nervous System Tissue on Beef Carcasses and in Comminuted Beef

We report the development and validation of a fluorescent enzyme-linked immunosorbent assay (ELISA) for glial fibrillary acidic protein (GFAP), which can be used as a rapid and sensitive method to detect CNS tissue in meat products. The fluorometric assay is sensitive to 0.2 ng GFAP and has an intra-assay coefficient of variation (CV) of 2.0% and an interassay CV of 14.1%. Bovine spinal cord and brain demonstrate dose-response curves that are parallel to GFAP standards, whereas peripheral sciatic nerve and cervical ganglia also cross-react at high tissue levels. The use of another central nervous system marker, syntaxin 1-B, was not effective for neural tissue detection. Less than 1.0 ng GFAP per mg tissue was found on most beef subprimals and advanced meat recovery (AMR) product. Occasional samples contained higher levels of GFAP, probably because of contamination by the carcass-splitting saw, incomplete removal of the spinal cord, or a chance sampling of a major nerve. Further reduction of CNS content was facilitated by removal of the cervical vertebrae and the spinal canal prior to processing beef chuck bones through AMR equipment. The presence of GFAP was very low (0.037 ng/mg) in beef patties collected from major processors throughout the USA. The presence of normal sausage ingredients or heating the product to 80 degrees C for 60 min did not affect the detection of GFAP. Heating the product to 115 degrees C for 100 min eliminated the detectability of GFAP.

Interventions to reduce microbiological contamination of beef variety meats

Hot water and solutions of acetic acid, lactic acid, or trisodium phosphate applied by immersion or spraying, chlorine solution applied by immersion, and exposure to steam in a pasteurization system, in a cabinet, or in combination with vacuum were evaluated for their effectiveness in reducing levels of bacterial contamination on samples of beef cheek meat, large intestine, lips, liver, oxtail, and tongue. Treated samples (five per treatment) and controls were analyzed for aerobic plate counts (APCs) on tryptic soy agar and for total coliform counts (TCCs) and Escherichia coli counts (ECCs) on Petrifilm. Acetic acid (2%) immersion and trisodium phosphate (12%) spraying and immersion for 10 s were among the most effective treatments in 16, 15, and 14, respectively, of 18 comparisons for reducing APCs, TCCs, and ECCs on four or more of the six variety meats tested. Acetic acid (2%) spraying, lactic acid (2%) immersion, and hot water (78 to 80 degrees C) spraying for 10 s were among the most effective treatments for reducing APCs, TCCs, and ECCs on four or more of the six variety meats. Chlorine (0.005%) immersion and steam were among the least effective treatments for reducing APCs, TCCs, and ECCs on variety meats. The results indicated that interventions applied to decontaminate beef carcasses can also be considered for decontamination of variety meats.

Potential for Disruption of Central Nervous System Tissue in Beef Cattle by Different Types of Captive Bolt Stunners

The application of pneumatic-powered air injection stunners (PPAISs), pneumatic-powered stunners (PPSs), and cartridge-fired stunners (CFSs) in commercial beef slaughter plants was evaluated to determine the extent of dissemination of central nervous system tissue. Fifteen beef slaughter plants in the western and central United States were visited to observe stunning methods and the condition of the hearts at postmortem inspection. As inspectors performed the normal opening of the hearts, the research observer evaluated the contents of the heart for the presence of clots and/or visible tissue segments in the right ventricle. In eight plants where PPAISs were used, 33% of hearts examined (n = 1,050) contained large clots in the right ventricles. In the four plants where CFSs were used, 1% of the hearts (n = 480) contained detectable clots. In three plants where the newly modified PPSs were used, 12% of the hearts (n = 450) contained detectable clots. Large segments of spinal cord were detected, collected, photographed, and confirmed histologically from two hearts in a plant that used a PPAIS. Most of the material was found in a single right ventricle and was composed of 10 to 13 cm segments of spinal cord.

Fate of Listeria monocytogenes in raw and cooked ground beef with meat processing additives

The effect of sodium lactate (1.8% w/w), sodium erythorbate (0.1% w/w), kappa-carrageenan (1% w/w), and the alginate meat binder (0.4% w/w, sodium alginate; 0.6% w/w lactic acid; and 0.075% w/w calcium carbonate) on Listeria monocytogenes survival and growth was determined in raw and cooked ground beef stored aerobically at 4 degrees C. There was no significant (P > 0.05) increase in numbers of L. monocytogenes during storage of raw ground beef. However, L. monocytogenes numbers were generally lower in treatments with sodium lactate, and higher in sodium erythorbate compared to controls and meat with other additives. Increases in total aerobic plate counts were less pronounced in raw meat formulated with sodium lactate and alginate meat binder than with other additives. Cooking meat with initial inoculum levels of 6.52 to 7.03 L. monocytogenes log CFU/g to 65 degrees C resulted in lower destruction (0.56 and 1.18 log CFU/g) in samples with added alginate meat binder and kappa-carrageenan, respectively, compared to the control. Survivors (2.11-3.73 log CFU/g) decreased initially and then increased slightly, but not significantly (P > 0.05), during storage (4 degrees C, 6 days) of the cooked products.

Attachment of Escherichia coli O157:H7 and Other Bacterial Cells Grown in Two Media to Beef Adipose and Muscle Tissues

Three strains of Escherichia coli O157:H7 were grown in tryptic soy broth (TSB) or in a sterile cattle manure extract at 35°C for 18 ± 2 h. Aliquots from both inocula containing 106 CFU/ml were used to inoculate 1-cm3 cubes of beef muscle or adipose tissue by immersion for 20 min at 21°C. After removal from the inoculum, one-half of the samples were analyzed for bacterial cell numbers and pH, and the other half were stored at 4°C for 2 or 3 h before analysis. Samples were analyzed by enumerating bacteria present in liquid droplets deposited on the tissue and bacteria loosely or strongly attached to the tissue in order to determine attachment strength. Total numbers of cells on beef muscle tissue (bacteria in liquid droplets, as well as those loosely and strongly attached) were 5.65 ± 0.14 and 5.76 ± 0.26 log CFU/cm2 for E. coli O157:H7 inocula grown in TSB and manure extract, respectively. The differences in attachment strength between inocula from the two media were not significant (P > 0.05). A 2-h storage period after exposure of muscle tissue to an E. coli O157:H7 inoculum did not influence attachment strength. Numbers of bacteria attached to adipose tissue and muscle (5.31 ± 0.08 and 5.48 ± 0.09 log CFU/cm2, respectively) were not significantly different (P > 0.05). After 3 h at 4°C, the attachment strength of E. coli O157:H7 cells on muscle or adipose tissue had not changed. Overall, the culture medium and type of beef tissue did not affect the numbers of E. coli O157:H7 cells attached, nor the strength of their attachment, to muscle or adipose tissue.

The effect of cooking temperature on the functional properties of beef proteins: The role of ionic strength, pH, and pyrophosphate.

This study examined the effect of ionic strength (0·12 to 0·52), pH (5·50 and 6·00), pyrophosphate (PP) concentration (0 and 0·31%) and cooking temperature (52 to 87°C) on the cook yield (CY) and tensile strength (TS) of beef homogenates. Increasing the ionic strength, pH and pyrophosphate concentration increased the temperature at which cooking loss first occurred and decreased the temperature required for maximum TS. For most treatments, ionic strengths between 0·32 and 0·42 prevented cooking loss at all temperatures; the lower ionic strengths were required at the higher pH and PP concentration. Maximum TS occurred at 66°C for treatments that had no cooking loss between 60° and 75°C. For treatments that had cooking loss in this temperature range, TS increased linearly with increasing temperature; however, the TS values of these treatments were much lower than those in the former category. CY and TS were optimized by heating to 66°C. PP had a positive effect on both functional properties at ionic strengths >0·25 but a negative effect at ionic strengths <0·25.