Verification of different glial fibrillary acidic protein (GFAP) analyses as accurate detectors of central nervous system tissue in advanced meat recovery (AMR) products.

A glial fibrillary acidic protein (GFAP) fluorescent enzyme linked immunosorbant assay (ELISA) was compared with an ELISA test kit for GFAP to determine the level of central nervous system (CNS) tissue in advanced meat recovery (AMR) products. The test kit results were highly correlated (r=0.975) with the fluorescent ELISA. Meat cuts and AMR were analyzed on site at 14 meat plants utilizing the test kits. In seven of the plants all AMR samples had less than 1 ng GFAP. Seven of the plants had greater than 1 ng GFAP in AMR samples. Development of proper process controls to eliminate inclusion of spinal cord in AMR materials should bring all values to less than 1 ng GFAP, a level slightly above background.

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.

Kinematics of a total arthroplasty of the ankle: comparison to normal ankle motion.

Although the concept of a total ankle arthroplasty has been advanced as a method for treating severe ankle arthritis, the clinical experience with all of the models developed has been discouraging. Both the constrained designs, which maximize joint contact area by restricting the available motion, and the unconstrained designs, which allow more normal motion at the expense of higher contact stresses, uniformly result in implant loosening, pain, and clinical failure in 2 to 7 years. This has led to the recommendation against the use of a total ankle arthroplasty except in very low-demand patients. Failure of ankle implants can be ascribed to either anatomic considerations (e.g.--the talus is too small to accommodate the stress transfers of a prosthesis), or mechanical etiologies. Abnormal 3-dimensional motion of the ankle following arthroplasty would fall into the latter category. This study examined the motion that occurs after implantation of an unconstrained-type total ankle arthroplasty. Using previously validated methodology, axially loaded ankle specimens were cycled through an arc of plantarflex/dorsiflexion while measuring the resulting coupled internal/external and varus/valgus rotations. The average coupled motions in prosthetic ankles were not significantly different than their intact controls. There was, however, a significantly increased amount of hysteresis (defined as the difference between the upper and lower pathways of coupled motion at any given sagittal position) that occurred as the ankle was dorsiflexed and plantar flexed. The increased hysteresis was seen in both the axial and coronal planes. This indicates that there was a greater permitted envelop of motion in the prosthetic ankles compared to normal ankles. It is hypothesized that this subtle change in ankle kinematics caused by the arthroplasty leads to abnormal stress transfer at the prosthesis-bone interface, thereby promoting early implant failure.

Injection of beef strip loins with solutions containing sodium tripolyphosphate, sodium lactate, and sodium chloride to enhance palatability.

Beef strip loins (46 U.S. Choice loins and 49 U.S. Select loins) were used to evaluate the potential for enhancing beef tenderness, juiciness, and flavor by injecting fresh cuts with solutions containing sodium tripolyphosphate, sodium lactate, and sodium chloride. One half of each loin served as an untreated control, and the other half was injected with either distilled water (110% of raw weight) or a solution containing phosphate/lactate/chloride solution (107.5, 110, 112.5, or 115% of raw weight). All phosphate/lactate/chloride solutions were formulated to produce injected product concentrations of .25% sodium tripolyphosphate, .5% sodium chloride, and 2.5% sodium lactate. Ten additional U.S. Select loins were injected to 110% of raw weight with a phosphate-only solution (final product concentration of .25% sodium tripolyphosphate) for comparison with Select loins injected to 110% with phosphate/lactate/chloride and with distilled water. Steaks from each control and treated loin section were cooked to two final internal temperatures (66 degrees C and 77 degrees C) for sensory panel evaluation and shear force measurement. Injection of subprimal cuts with phosphate/lactate/chloride solutions improved tenderness (P < .05), juiciness (P < .05), and cooked beef flavor (P < .10) of strip loin steaks and was especially effective for maintaining tenderness and juiciness of steaks cooked to the higher final internal temperature. Injection of Select loins with a solution containing only sodium tripolyphosphate was not effective for improving beef tenderness or juiciness and tended to impart off-flavors characterized by sensory panelists as soapy and sour. Injection of fresh cuts with phosphate/lactate/chloride solutions could assist the beef industry's efforts to improve product quality and consistency.

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.

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.

Growth and fresh meat quality characteristics of pigs supplemented with vitamin E.

Crossbred pigs (n = 30) were fed to determine the influence of supplementation with vitamin E on growth and slaughter characteristics of swine and on the quality characteristics of fresh pork. Pigs received either a control diet containing no vitamin E (CON) or a diet formulated to contain 100 mg of vitamin E/kg feed (VITE). During 84 d of feeding, feed intake and weight gain were measured every 2 wk. After the feeding period, pigs were slaughtered and the loin from the left side of each carcass was removed 4 d after death. Alpha-Tocopherol concentration and proximate composition of the longissimus muscle were determined. Loins were sliced into 10-cm sections and stored under vacuum (2 degrees C) for 0, 14, 28, and 56 d. After storage, loins were sliced into 2.54-cm chops, wrapped in polyvinyl chloride film and stored in a retail case (2 to 4 degrees C) for 5 d. Thiobarbituric acid (TBA) values, Hunter L, a, and b values, total plate counts, pH, purge loss, drip loss, cook loss, taste panel characteristics, and visual panel characteristics were evaluated. Growth traits, slaughter characteristics, and proximate composition did not differ (P > .05) between dietary treatment groups. Alpha-Tocopherol concentrations were greater (P < .05) and TBA values during extended retail display were less (P < .05) for VITE chops than for CON chops. Overall palatability ratings were more desirable (P < .05, at 14 d of vacuum storage) for VITE chops than for CON chops. Color measurements, sensory characteristics, total plate counts, pH, purge loss, drip loss, and cook loss were not influenced (P > .05) by vitamin E supplementation. These results indicated that at the tissue alpha-tocopherol concentrations of the present study, vitamin E supplementation of the growing-finishing diet of hogs reduced lipid oxidation in fresh pork but did not influence pork color or tissue drip loss.

Injection-site lesions: incidence, tissue histology, collagen concentration, and muscle tenderness in beef rounds.

The national incidence and extent of injection-site lesions in the muscles of the round were determined via audits conducted at retail stores and in purveying establishments. Two additional experiments were conducted to examine the subsequent effects of pharmaceutical administration on tissue histology, soluble and insoluble collagen concentration, and muscle tenderness in beef bottom-rounds. Injection-site lesion incidence in beef round cuts audited at retail (n = 3,538) and in steak-cutting facilities (n = 15,464) was 8.45 and 10.04%, respectively, with an average lesion-trim of 314.7 and 191.59 g, respectively, in these two studies. Lesion classification revealed that 93.20 and 99.91% of lesions reported for the retail and purveyor audits, respectively, were chronologically aged lesions. Overall, 19,002 round cuts were examined, and injection-site lesion incidence (nationally) was 9.74%, whereas lesion-trim averaged 211.8 g. Warner-Bratzler shear measurements taken near lesions and in areas 7.62 cm from the lesions were higher (P < .001) for lesioned, than for control bottom-round steaks. Warner-Bratzler shear values for lesion cores were 3.5 times greater than those in paired control (non-affected) steaks. Concentrations of insoluble and soluble collagen were much higher (P < .001) at the site of the lesion center in lesion-afflicted vs control steaks. Histological determinations of the relative proportions of muscle, connective tissue and fat to a distance of 5.08 cm from the site of the lesion center confirmed that severe disruption of muscle tissue constituents and architecture had occurred. Injection-site lesions occur at an unacceptable frequency in the muscles of the round, and severe tissue changes accompany these lesions that can dramatically affect tenderness of those cuts.

Incidence of injection-site blemishes in beef top sirloin butts.

The occurrence of damaged muscle tissue resulting from intramuscular injections of animal-health products represents a "quality control" problem and an economic loss to the beef industry. Five individual and sequential national audits of injection-site blemishes in beef top sirloin butts have been conducted at the steak-cutter level. During the most recent audit (March 1993), the incidence of injection-site blemishes in top sirloin butts was determined to be 10.87 +/- 2.99%, with an average weight per blemish of 123.39 +/- 5.48 g. A 5-point classification system used to partition the blemishes into chronological stages of the healing process suggested that the majority of the blemishes originated at the cow-calf or stocker levels, or early in the finishing period. Evaluation of blemish data by geographic location of plant-of-origin suggests that the problem occurs throughout the beef production sector.

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.

Growth of Listeria monocytogenes Inoculated in Waste Fluids Collected from a Slaughterhouse.

Cells of Listeria monocytogenes Scott A were artificially inoculated (26-378 CFU/ml) in waste fluids collected from a lamb slaughtering operation, and their growth was determined at 8 and 35°C. Samples tested included residual fluid from carcass rinsing (after hide and offal removal); a mixture of waste fluids from the floor (collected during the entire slaughtering and washing process); and a standard floor drain sample (drawn from the floor drain 5 min after clean-up was complete). Growth of L. monocytogenes within 24 h of incubation of the various waste fluids was more rapid at 35 (1-4 logs) than at 8°C (0.5-2 logs). Average generation times of L. monocytogenes growth at 8°C were in the range of 12.3 to 35.1 h, depending on waste fluid, while at 35°C generation times were in the range of 1.1-6.4 h. Therefore, waste fluids in slaughterhouse facilities can support growth of L. monocytogenes , which may then be introduced on carcasses and retail meat cuts. Thus, slaughterhouse temperatures should be as low as possible and cleaning and sanitation should be frequent in order to minimize carcass and retail meat contamination with the pathogen.

Gastric ulcers in horses: a comparison of endoscopic findings in horses with and without clinical signs.

Gastroendoscopic examinations were performed on 187 horses, ranging from one to 24 years. Eighty-seven horses had clinical problems including chronic, recurrent colic for seven or more days (25), one or more episodes of colic within the previous seven days (13), or acute colic (10), diminished appetite (53), poor bodily condition (40), and/or chronic diarrhoea (9). One hundred horses that had no signs of gastrointestinal problems were examined as part of a gastroendoscopic survey. Lesions observed in the squamous fundus, squamous mucosa adjacent to the margo plicatus along the greater curvature, glandular fundus, and the squamous mucosa along the lesser curvature were scored on a scale of 0-4, with 0 representing no lesions and 4 representing the most severe lesions. The mean endoscopic scores for the squamous fundus, margo plicatus and lesser curvature were significantly greater (P < 0.001) in horses with clinical signs than those without signs. This was because of the greater number of horses with lesions in the symptomatic group (80/87) compared to those without signs (52/100), and the greater severity of lesions in the horses with clinical signs. Of the horses, 74 were in race training. There was a significantly (P < 0.01) greater prevalence and severity of lesions at all sites except the glandular fundus in horses in training compared to those not in training, and in the horses in training with clinical signs (n = 37) compared to those in training without clinical signs (n = 37).

Transdermal clonidine compared with hydrochlorothiazide as monotherapy in elderly hypertensive males.

Sixteen of 22 elderly male patients (aged 60-74 years) who had previously taken only hydrochlorothiazide 50 mg completed a study evaluating the safety, efficacy, and tolerability of 12-20 weeks of transdermal clonidine (Catapres TTS) as monotherapy for mild hypertension. Thirteen of the sixteen patients (81%) responded to transdermal clonidine which was begun after 28 days of placebo. Five patients discontinued transdermal clonidine therapy because of intolerable skin irritation, and one because of daytime fatigue. Clonidine caused none of the metabolic effects we observed with hydrochlorothiazide: no change in serum potassium, uric acid, cholesterol, or triglyceride. Eleven of the 22 patients (50%) who began the study experienced a skin reaction under the transdermal clonidine patch. The incidence of dry mouth and fatigue in patients using transdermal clonidine was dose-related and similar to reports of dry mouth and fatigue in patients taking oral clonidine tablets. Rebound hypertension occurred in one patient upon withdrawal of transdermal clonidine. There was no effect of transdermal clonidine or hydrochlorothiazide on cognitive function or emotional state tested with three questionnaires. Overall, transdermal clonidine, in various doses, was as effective as hydrochlorothiazide in elderly male hypertensive patients. The effectiveness of both was inversely proportional to the level of untreated blood pressure. The high incidence of skin reactions limited prolonged use of transdermal clonidine in our patients.

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.

Home blood pressure readings in clinical hypertension research.

The feasibility and reliability of home blood pressure measurement in clinical research were determined as part of a study to document the safety and efficacy of two captopril dosage regimens. Twenty patients were studied. Patients were instructed to record blood pressures twice daily (before morning and evening doses of captopril) for 24 weeks. All patients were instructed in blood pressure measurement and were required to demonstrate the accuracy of their techniques. Supine and standing blood pressures were measured every two weeks in the clinic. The mean of all home blood pressures recorded in the two-week period before each clinic visit was compared with the measurement obtained at that clinic visit for each patient. Blood pressure data could be evaluated for 17 patients. Each patient recorded 330-380 readings, and the mean home blood pressure measurement for each two-week period was calculated from 26-32 separate readings. Home blood pressures were consistently lower than clinic measurements but showed similar trends in response to treatment. There was good correlation between values obtained at home and in the clinic. Morning and evening blood pressures obtained by patients were not significantly different. Home blood pressure monitoring is a useful tool in hypertension research.