Effects of housing and feeding systems on performance of neonatal Holstein bull calves.

As the dairy industry continues to grow, more dairy calves are available for dairy, beef, and veal purposes. Rearing systems must be highly efficient to make this industry cost efficient, making the evaluation of rearing methods important to establish the most practical method. A study was designed and conducted to evaluate effects of housing and feeding systems on performance of neonatal Holstein bull calves. Treatments (2 × 2 factorial arrangement) consisted of: 1) individually housed, bottle-fed (n = 5 bull calves); 2) individually housed, bucket-fed (n = 5 bull calves); 3) group-housed, bottle-fed (n = 5 pens; 4 bull calves/pen); and 4) group-housed, bucket- (trough) fed (n = 5 pens; 3 or 4 bull calves/pen). Feeding treatments began on d 7 when calves had been acclimated to their new environment. Body weight measurements were collected every 7 d and blood samples were collected on d 0, 28, 55, and 66 for ß-hydroxybutyrate (BHBA) concentration as a gross indicator of ruminal development. No housing × feeding interactions or feeding treatment effects were observed (P > 0.10). Average DMI (dry feed plus milk replacer) was increased (P < 0.05) for group-housed vs. individual animals after d 41, and final BW was greater (P < 0.05) for group-housed calves compared with individually housed calves. Feed efficiency and ADG, however, remained similar (P > 0.10) for all treatments. Fecal scores (P > 0.26), CV for BW (P > 0.26), and BHBA concentrations (P > 0.14) showed no differences among treatments. Housing system had greater effect on calf performance compared with milk feeding regimen.

Effect of two dietary concentrate levels on tenderness, calpain and calpastatin activities, and carcass merit in Waguli and Brahman steers.

The objective of this study was to compare carcass characteristics of a newly introduced breed, the Waguli (Wagyu x Tuli), with the carcass characteristics of the Brahman breed. Brahman cattle are used extensively in the Southwest of the United States because of their tolerance to adverse environmental conditions. However, Brahman carcasses are discounted according to the height of their humps because of meat tenderness issues. The Waguli was developed in an attempt to obtain a breed that retained the heat tolerance of the Brahman but had meat quality attributes similar to the Wagyu. Twenty-four animals were used. Six steers from each breed were fed a 94% concentrate diet and 6 steers from each breed were fed an 86% concentrate diet. Eight steers, 2 from each group, were harvested after 128 d, after 142 d, and after 156 d on feed. Waguli steers had larger LM, greater backfat thickness, greater marbling scores, and greater quality grades than the Brahman steers (P < 0.05). The Japanese Wagyu breed is well known for its highly marbled and tender meat, and these traits are also present in the Waguli. The Waguli had significantly lower Warner-Bratzler shear force values than the Brahman steers after 7 and 10 d of postmortem aging (P < 0.05); this difference decreased after 14 d postmortem (P = 0.2), when tenderness of the slower aging Brahman had increased to acceptable levels. Toughness of the Brahman has been associated with high levels of calpastatin in Brahman muscle, and the Waguli LM had significantly less calpastatin activity (P = 0.02) at 0 h postmortem than the Brahman LM. At 0-h postmortem, the total LM calpain activity did not differ between the Brahman and Waguli (P = 0.57). Neither diet nor days on feed had any significant effect on the 0-h postmortem calpain or at 0-h postmortem calpastatin activity, nor an effect on Warner-Bratzler shear-force values. In conclusion, LM muscle from the Waguli steers had a high degree of marbling, lower shear force values, and low calpastatin activity, all of which are related to more tender meat.

Isolation and characterization of mu-calpain, m-calpain, and calpastatin from postmortem muscle. I. Initial steps.

Evidence has indicated that mu-calpain, m-calpain, and calpastatin have important roles in the proteolytic degradation that results in postmortem tenderization. Simple assays of these 3 proteins at different times postmortem, however, has shown that calpastatin and mu-calpain both rapidly lose their activity during postmortem storage, so that proteolytic activity of mu-calpain is nearly zero after 3 d postmortem, even when assayed at pH 7.5 and 25 degrees C, and ability of calpastatin to inhibit the calpains is 30% or less of its ability when assayed at death. m-Calpain, however, retains much of its proteolytic activity during postmortem storage, but the Ca(2+) requirement of m-calpain is much higher than that reported to exist in postmortem muscle. Consequently, it is unclear how the calpain system functions in postmortem muscle. To clarify this issue, we have initiated attempts to purify the 2 calpains and calpastatin from bovine semitendinosus muscle after 11-13 d postmortem. The known properties of the calpains and calpastatin in postmortem muscle have important effects on approaches that can be used to purify them. A hexyl-TSK hydrophobic interaction column is a critical first step in separating calpastatin from the 2 calpains in postmortem muscle. Dot-blot assays were used to detect proteolytically inactive mu-calpain. After 2 column chromatographic steps, 5 fractions can be identified: 1) calpastatin I that does not bind to an anion-exchange matrix, that does not completely inhibit the calpains, and that consists of small polypeptides <60 kDa; 2) calpastatin II that binds weakly to an anion-exchange matrix and that contains polypeptides <60 kDa; all these polypeptides are smaller than the native 115- to 125-kDa skeletal muscle calpastatin; 3) proteolytically active mu-calpain even though very little mu-calpain activity can be detected in zymogram assays of muscle extracts from 11- to 13-d postmortem muscle; this mu-calpain has an autolyzed 76-kDa large subunit but the small subunit consists of 24-, 26- and a small amount of unautolyzed 28-kDa polypeptides; 4) proteolytically active m-calpain that is not autolyzed; and 5) proteolytically inactive mu-calpain whose large subunit is autolyzed to a 76-kDa polypeptide and whose small subunit contains polypeptides similar to the proteolytically active mu-calpain. Hence, loss of calpastatin activity in postmortem muscle is due to its degradation, but the cause of the loss of mu-calpain activity remains unknown.

Effect of postmortem storage on activity of mu- and m-calpain in five bovine muscles.

An in situ system involving incubation of 60- to 80-g pieces of muscle at 4 degrees C under different conditions was used to determine the effects of time of postmortem storage, of pH, and of temperature on activities of mu- and m-calpain activity in bovine skeletal muscle. Casein zymograms were used to allow measurement of calpain activity with a minimum of sample preparation and to ensure that the calpains were not exposed to ionic strengths of 100 or greater before assay of their activities. In 4 of the 5 muscles (longissimus dorsi, lumbar; longissimus dorsi, thoracic; psoas major; semimembranosus; and triceps brachii) studied, mu-calpain activity decreased nearly to zero within 48 h postmortem. Activity of m-calpain also decreased in the in situ system used but at a much slower rate. Activities of both mu- and m-calpain decreased more slowly in the triceps brachii muscle than in the other 4 muscles during postmortem storage. Although previous studies have indicated that mu-calpain but not m-calpain is proteolytically active at pH 5.8, these studies have used calpains obtained from muscle at death. Both mu- and m-calpain are proteolytically inactive if their activities are measured at pH 5.8 and after incubating the muscle pieces for 24 h at pH 5.8. Western analysis suggested that neither the large 80-kDa subunit nor the small 28-kDa subunit of m-calpain was autolyzed during postmortem storage of the muscle pieces. As has been reported previously, the 80-kDa subunit of mu-calpain was autolyzed to 78- and then to a 76-kDa polypeptide after 7 d postmortem, but the 28-kDa small subunit was not autolyzed; hence, the autolyzed mu-calpain molecule in postmortem muscle is a 76-/28-kDa molecule and not a 76-/18-kDa molecule as previously assumed. Because both subunits were present in the postmortem calpains, loss of mu-calpain activity during postmortem storage is not due to dissociation of the 2 subunits and inactivation. Although previous studies have shown that the 76-/18-kDa mu-calpain molecule is completely active proteolytically, it is possible that the 76-/28-kDa mu-calpain molecule in postmortem muscle is proteolytically inactive and that this accounts for the loss of mu-calpain activity during postmortem storage. Because neither mu- nor m-calpain is proteolytically active at pH 5.8 after being incubated at pH 5.8 for 24 h, other proteolytic systems such as the caspases may contribute to postmortem proteolysis in addition to the calpains.

DairyBeef: maximizing quality and profits--a consistent food safety message.

To respond to meat safety and quality issues in dairy market cattle, a collaborative project team for 7 western states was established to develop educational resources providing a consistent meat safety and quality message to dairy producers, farm advisors, and veterinarians. The team produced an educational website and CD-ROM course that included videos, narrated slide sets, and on-farm tools. The objectives of this course were: 1) to help producers and their advisors understand market cattle food safety and quality issues, 2) help maintain markets for these cows, and 3) help producers identify ways to improve the quality of dairy cattle going to slaughter. DairyBeef. Maximizing Quality & Profits consists of 6 sections, including 4 core segments. Successful completion of quizzes following each core segment is required for participants to receive a certificate of completion. A formative evaluation of the program revealed the necessity for minor content and technological changes with the web-based course. All evaluators considered the materials relevant to dairy producers. After editing, course availability was enabled in February, 2003. Between February and May, 2003, 21 individuals received certificates of completion.

Properties of myofibril-bound calpain activity in longissimus muscle of callipyge and normal sheep.

Properties of the calpain bound to myofibrils in longissimus muscle from callipyge or noncallipyge sheep were examined after 0, 1, 3, and 10 d of postmortem storage at 4 degrees C. Western analysis has shown that most of this calpain is mu-calpain, although the sensitivity of the antibodies used in the earlier studies could not eliminate the possibility that up to 10% of the calpain was m-calpain. The calpain is bound tightly, and very little is removed by washing with the detergent Triton X-100; hence, it is not bound to phospholipids in the myofibril. Over 25% of total mu-calpain was bound to myofibrils from at-death muscle, and this increased to approximately 40% after 1 d postmortem. The amount of myofibril-bound mu-calpain increased only slightly between 1 and 10 d of postmortem storage. The percentage of autolyzed mu-calpain increases with time postmortem until after 10 d postmortem, when all myofibril-bound mu-calpain is autolyzed. The specific activity of the myofibril-bound calpain is very low and is only 6 to 13% as high as the specific activity of extractable mu-calpain from the same muscle. It is unclear whether this low specific activity is the result of unavailability of the active site of the myofibril-bound calpain to exogenous substrate. The myofibril-bound calpain degrades desmin, nebulin, titin, and troponin T in the myofibrils, and also releases undegraded alpha-actinin and undergoes additional autolysis when incubated with Ca2+; all these activities occurred slowly considering the amount of myofibril-bound calpain. Activity of the myofibril-bound calpain was partly (58 to 67%) inhibited by the calpain inhibitors, E-64 and iodoacetate; was more effectively inhibited by a broader-based protease inhibitor, leupeptin (84 to 89%); and was poorly inhibited (43 to 45%) by calpastatin. Release of undegraded alpha-actinin and autolysis are properties specific to the calpains, and it is unclear whether some of the myofibril-bound proteolytic activity originates from proteases other than the calpains or whether the active site of myofibril-bound calpain is shielded from the inhibitors. Activities and properties of the myofibril-bound calpain were identical in longissimus muscle from callipyge and normal sheep, although previous studies had indicated that the "normal" longissimus was much more tender than the callipyge longissimus. Hence, it seems unlikely that the myofibril-bound calpain has a significant role in postmortem tenderization of ovine longissimus.

The calpain system in three muscles of normal and callipyge sheep.

Activities of mu- and m-calpain and of calpastatin were measured at four different times during postmortem storage (0, 1, 3, and 10 d) in three muscles from either callipyge or noncallipyge (normal) sheep. The weights of two muscles, the biceps femoris and the longissimus, are greater in the callipyge phenotype, whereas the weight of the infraspinatus is not affected. The activity of m-calpain was greater (P < 0.05) in the biceps femoris and longissimus from callipyge than in those from normal sheep, but it was the same in the infraspinatus in the two phenotypes. The extractable activity of m-calpain did not change (biceps femoris and infraspinatus) or decreased slightly (longissimus) during postmortem storage. Extractable activity of mu-calpain decreased to zero or nearly zero after 10 d postmortem in all muscles from both groups of sheep. The rate of decrease in mu-calpain activity was the same in muscles from the callipyge and normal sheep. At all time points during postmortem storage, calpastatin activity was greater (P < 0.05) in the biceps femoris and longissimus from the callipyge than from the normal sheep, but it was the same in the infraspinatus from callipyge and normal sheep. Calpastatin activity decreased (P < 0.05) in all three muscles from both phenotypes during postmortem storage; the rate of this decrease in the callipyge biceps femoris and longissimus and in the infraspinatus from both the callipyge and normal sheep was slow, especially after the first 24 h postmortem, whereas calpastatin activity in the biceps femoris and longissimus from the normal sheep decreased rapidly. During postmortem storage, the 125-kDa calpastatin polypeptide was degraded, but the 80-kDa subunit of mu-calpain was cleaved only to 76- and 78-kDa polypeptides even though extractable mu-calpain activity declined nearly to zero. Approximately 50 to 60% of total mu-calpain became associated with the nonextractable pellet after 1 d postmortem. The myofibril fragmentation index for the biceps femoris and longissimus from normal sheep increased significantly during postmortem storage. The fragmentation index for the infraspinatus from the callipyge and normal sheep increased to an intermediate extent, whereas the index for the biceps femoris and longissimus from the callipyge did not change during 10-d postmortem storage. The results suggest that postmortem tenderization is related to the rate of calpastatin degradation in postmortem muscle and that calpastatin inhibition of the calpains in postmortem muscle is modulated in some as yet unknown manner.

Evaluation of calpastatin activity measures in ante- and postmortem muscle from half-sib bulls and steers.

Calpastatin activity measured at 24 h postmortem in bovine longissimus muscle (PMLD24) is correlated with Warner-Bratzler shear force (WBS) measurements, an objective measure of tenderness. A live-animal measurement of calpastatin activity that correlates with 24-h postmortem activity would provide information for selection programs without the expense of progeny testing. The purpose of this study was to evaluate the effectiveness of calpastatin activity measurements obtained on tissue samples from live animals and to determine the relationship among various calpastatin activity measures and tenderness determined by WBS and sensory panel. Biopsies (approximately 10 g) were obtained surgically 2 d before slaughter from the supraspinatus muscle on the anterior surface of the scapula (LISH0) from contemporary purebred Angus bulls (n = 12) and steers (n = 17). Biopsies from a subset of these cattle (n = 12) were refrigerated at 4 degrees C to simulate the postmortem cooling process for 24 h (LISH24) prior to extraction. A rib section anterior to the 12 and 13th rib interface was collected from all animals at the commercial abattoir between 22 and 23 h postmortem for PMLD24, sensory panel, and WBS measurements. A postmortem shoulder muscle sample (PMSH24) was collected at the same time. Calpastatin was extracted from all muscle samples using a heated calpastatin activity protocol. Sensory panel tenderness, WBS, LISH0, LISH24, and PMSH24 were not different between bulls and steers. However, PMLD24 values were significantly different. Significant partial correlations were found between WBS and sensory panel tenderness (-.55), between WBS and PMLD24 (-.43), and between LISH24 and PMLD24 (.78). Therefore, similar calpastatin activity values are possible with ante- and postmortem tissue samples, suggesting the possibility of using measurements from live-tissue biopsies from other than the longissimus muscle to predict end product tenderness.

Effect of freezing and microbial growth on myoglobin derivatives of beef.

The effect of freezing and bacterial growth on the discoloration of beef was assessed by measuring myoglobin derivatives myoglobin (MB), oxymyoglobin (MBO(2)), and metmyoglobin (METMB) on the surfaces of fresh and frozen-thawed packaged beef cuts stored at 2 degrees C and analyzed after 0, 3, 6, 9, and 12 days of storage. MB, MBO(2), and METMB concentrations were measured spectrophotometrically. Frozen-thawed beef samples experienced less "blooming" (conversion of MB to MBO(2)) and more rapid discoloration than fresh cuts during storage. By day 3, >20% METMB was formed in the frozen-thawed samples, whereas the fresh samples reached this value after day 6 of storage. The rates of MB oxidation were similar (P > 0.05) for sterile and frozen-thawed inoculated (Pseudomonas fluorescens at a rate of 1.5 colony forming units/cm(2).cm(2) area) samples from day 0 through day 6 of storage. For storage periods of less than a week, bacterial growth is not a major cause of meat discoloration. After day 6, the high bacterial growth rate resulted in a rapid increase in METMB formation. Possible mechanisms for MB oxidation in frozen-thawed beef are suggested.

Bovine lipoprotein and apolipoprotein profiles as influenced by sex and growth.

Three (intact) Angus males and females that were half-sibs and born within 21 d of each other were selected for this study. Each animal was bled periodically from birth to slaughter (18 mo) to determine the qualitative composition of plasma lipoproteins and apolipoproteins during its growth and development. Major components observed were: 1) very low-density (VLDL), 2) low-density (LDL), and 3) high-density lipoproteins (HDL). Individual amounts of triglycerides, cholesterol, and proteins for the VLDL were not different (P greater than .05) between sexes at any time during growth and development. At 1 yr of age and 15 mo of age, females had significantly larger (143.4 and 93.5 mg/dl) amounts of protein in the HDL than males (67.0 and 93.5 mg/dl), respectively. Within the male group, the LDL triglyceride concentration of calves was significantly (P less than .05) higher (7.4 mg/dl) than at all other bleeding times. Within the female group, cholesterol values for the VLDL were significantly (P less than .05) larger as calves and weanlings (16.5 and 21.7 mg/dl respectively) than for other bleeding periods. At all stages of growth and development, the HDL apoprotein profiles showed a distinct band with a weight of about 28,000 Da, which represented apolipoprotein-A-I. During the suckling stage, pooling of LDL fractions provided two components on the acrylamide gel (7.5 to 20%), apolipoprotein-B and a low molecular weight band. At 12 and 15 mo, no low molecular weight band was present in the pooled LDL fraction.(ABSTRACT TRUNCATED AT 250 WORDS)

Apolipoprotein E-rich HDL binding to liver plasma membranes in copper-deficient rats.

Copper deficiency in rats raises plasma cholesterol concentration while reducing live cholesterol concentration. One consequence of this cholesterol redistribution is the accumulation of a large high-density lipoprotein (HDL) particle rich in apolipoprotein E (apo E). The purpose of this study was to determine, using an in vitro binding assay, if the interaction of apo E-rich HDL with hepatic lipoprotein binding sites may be affected by copper deficiency. Male Sprague-Dawley rats were divided into two dietary treatments (copper-deficient and -adequate) and placed on a dietary regimen for 8 weeks. Subsequent to exsanguination, hepatic plasma membranes were prepared and apo E-rich HDL was isolated from rats of each treatment by ultracentrifugation, agarose column chromatography, and heparin-Sepharose affinity chromatography. Total binding and experimentally derived specific binding of 125I-apo E-rich HDl to hepatic plasma membranes indicated greater binding when lipoproteins and membranes from copper-deficient animals were used in the assay compared to controls. Scatchard analysis of specific binding data indicated that equilibrium binding affinity (Kd) was also affected by copper deficiency. The hepatic binding sites recognizing apo E-rich HDL were not affected by EDTA or pronase, of relatively high capacity, and recognized a variety of other rat lipoproteins.

Lipoprotein receptors in copper-deficient rats: apolipoprotein E-free high density lipoprotein binding to liver membranes.

Twenty-four male weanling Sprague-Dawley rats were randomly and equally divided into two dietary treatments, copper-deficient and adequate (0.7 mg and 8.0 mg Cu/kg diet, respectively). Deionized water and diet were provided ad libitum. After 8 weeks, rats were exsanguinated, membranes prepared from livers, and plasma high density lipoproteins (HDL) isolated by ultracentrifugation and agarose column chromatography. Heparin-sepharose affinity chromatography was used to isolate subfractions of HDL devoid of apolipoprotein E (apo E-free HDL). The apo E-free HDL derived from rats of each dietary treatment were iodinated and bound to liver membranes prepared from rats of both treatment groups. Total binding data, specific binding data, and computer derived estimates of maximum equilibrium binding (Bmax) indicate less binding was observed when lipoproteins and membranes from copper-deficient animals were used in the binding assay compared to controls. In addition, a 2 X 2 factorial analysis of binding parameters derived from all experiments demonstrated a significant lipoprotein effect, indicating the reduction in binding may be associated with apo E-free HDL obtained from copper-deficient rats. The present findings suggest a reduction in binding of apo E-free HDL to their binding sites may contribute to the hypercholesterolemia and hyperlipoproteinemia observed in copper deficiency.

Growth of Aerobic Psychrotrophs and Color Changes of Precooked Beef Slices as Affected by Packaging Procedure 1.

Precooked beef slices from top round roasts were used in replicate trials to determine the effects of packaging treatment upon shelf-life characteristics (microbial growth and color attributes) during retail storage. Roasts were dry-roasted to an internal temperature of 60°C, cooled for 1 h, then sliced (3-4 mm). Slices were packaged in: (a) vacuum; (b) 15% CO2, 40% O2, 45% N2; (c) 15% CO2, 20% O2, 65% N2; or (d) 15% CO2, 10% O2, 75% N2 and held at 4°C for up to 18 d. Enumeration of microorganisms and determination of color attributes were done at 0, 3, 6, 9, 12, 15, and 18 d of storage. Psychrotrophic counts were higher (P<.05) on slices stored 9-18 d in atmospheres containing ambient (20%) oxygen level as compared to the other packaging treatments. Vacuum packaging exhibited a greater lag phase for psychrotrophs. Vacuum packaged slices maintained a superior cooked beef color for 18 d and had greater (P<.05) chroma readings, indicating a greater color intensity.

Microbial Changes of Precooked Beef Slices as Affected by Packaging Procedure.

Precooked beef slices from top round roasts were used in replicate trials to determine the effects of packaging treatment upon microbial growth during retail storage. Roasts were dry roasted to an internal temperature of 60°C, cooled for 1 h, then sliced (3 to 4 mm) and packaged in vacuum or an atmosphere containing 15% CO2/40% O2/45% N2. Slices were stored either at 2, 6 or 10°C for up to 21 d. Enumeration of psychrotrophs, mesophiles, thermophiles and molds was determined after 0, 7, 14 and 21 d of storage. At 6 and 10°C storage, psychrotrophic organisms did not increase (P<0.05) on vacuum packaged beef slices during the 21-d storage period, but did increase (P<0.05) on slices stored in the gas mixture. Conversely, at 2°C storage, psychrotrophs increased (P<0.05) in vacuum at day 21 but not under gas atmosphere storage. Mesophiles did not increase significantly at 2 or 6°C storage within either packaging treatment during 21 d of storage. Mold growth did not occur on slices stored at 2°C.

Effect of Different Packaging Treatments on Microbiological and Sensory Evaluation of Precooked Beef Roasts.

Thirty boneless top round roasts were used in each of two trials to determine the effects of various packaging treatments on precooked roast beef acceptability. Roasts were dry roasted to an internal temperature of 60°C, cooled for 1 h and packaged by one of three methods: (a) vacuum packaging, (b) packaging in 100% CO2 atmosphere or (c) packaging in 15% CO2:30% O2:55% N2 atmosphere. Roasts were held at 4°C for up to 21 d. Enumeration of mesophiles and psychrotrophs, sensory evaluations and shrinkage percentages were determined at 0, 7, 14 and 21 d of storage to evaluate acceptability. Counts of mesophiles and psychrotrophs from 100% CO2-treated roasts were significantly lower (P<0.05) than counts from vacuum-packaged roasts after 14 and 21 d of storage, whereas counts from gas mixture-treated roasts were not (P>0.05) different. After 7 d of storage, microbial numbers were similar, regardless of treatment. Sensory evaluation analyses showed that vacuum-packaged roasts exhibited little deterioration of quality characteristics at 21 d of storage, whereas both gas-treated roasts demonstrated quality deterioration by 14 and 21 d of storage. Vacuum-packaged roasts were preferred by panelists at 14 and 21 d, but CO2-treated roasts were preferred at 7 d. No treatment effects were evident upon shrinkage until 21 storage days. At 21 d, vacuum-packaged roasts exhibited the lowest moisture loss.

Impaired glucose tolerance in copper-deficient rats.

Forty-eight male weanling Sprague-Dawley rats were randomly and equally divided into two dietary treatments (copper-deficient and adequate: 0.85 mg and 8 mg of Cu/kg diet). Deionized water and diet were provided ad libitum. After 5 weeks, the rats were fasted for 18 hours, anesthetized with sodium pentobarbitol and injected intravenously with glucose (1 g/kg body wt in a 50% wt/vol solution). Six rats from each treatment were killed by exsanguination at 0, 30, 60 and 120 minutes after glucose injection. Liver copper was measured by atomic absorption spectrophotometry. Reduction in liver copper content and elevation in heart weight confirmed that the rats fed the test diet were copper-deficient. Plasma glucose levels in copper-deficient rats were significantly higher at 30 and 60 minutes compared to controls. After 2 hours there were no significant differences between treatments. Plasma insulin levels measured by radioimmunoassay were significantly lower at 30 minutes, but higher at 60 and 120 minutes in rats fed the test diet as compared to controls. It would thus appear likely that copper deficiency interferes with normal glucose utilization.

Growth of Staphylococcus aureus on Beef Steaks as Influenced by Type of Packaging 1, 2.

Beef loin steaks were inoculated with 105 cells of Staphylococcus aureus /cm2 in two separate trials to determine the effects of different packaging treatments upon the organism's growth. Each trial utilized 72 samples which were randomly assigned to four packaging treatments: (1) over-wrapped in oxygen-permeable film; (2) vacuum packaged; (3) packaged in barrier bags flushed with a 60% CO2 : 40% O2 gas atmosphere then evacuated and sealed; and (4) packaged in barrier bags filled with a 60%CO2 : 40% O2 gas atmosphere. In each treatment group, 12 samples were inoculated with S. aureus while 6 were uninoculated and used as controls. All samples were displayed under simulated retail conditions at 10 C and enumerated for S. aureus after 3, 6 and 9 days. Numbers of S. aureus remained relatively constant for all treatments throughout the 9 day period. Results from a comparison of treatments for Trials I and II were variable; however, no significant treatment effects were found when data from both trials were pooled.

Growth of Salmonella typhimurium and Mesophilic Organisms on Beef Steaks as Influenced by Type of Packaging 1, 2.

Two trials, each utilizing 72 samples of fresh beef loin steak, were done to determine the effects of various packing systems upon growth of Salmonella typhimurium . Samples were inoculated with 105 cells/cm2 of the organism and randomly assigned to four packaging treatments: (1) overwrapping in oxygen-permeable film; (2) vacuum packaging; (3) packaging in barrier bags flushed with a 60% CO2: 40% O2 gas atmosphere then evacuated and sealed; and (4) packaging in barrier bags filled with a 60% CO2: 40% O2 gas atmosphere. Twelve steak samples were inoculated with S. typhimurium and 6 were uninoculated and served as a control in each treatment group. Samples were displayed in retail meat cases at 10 C for 3, 6 or 9 days, when they were evaluated for shrinkage and numbers of mesophilic organisms and S. typhimurium . Percent shrinkage was not affected (P>0.05) by packaging treatment. Counts of mesophilic organisms were similar (P>0.05) for vacuum- and gas-treated steaks, which were significantly lower (P<0.05) than counts from film overwrapped samples. Numbers of S. typhimurium increased significantly (P<0.05) during storage on samples wrapped with oxygen permeable film but remained low and fairly constant for vacuum- or gas-treated steaks. After 9 days of display, the film overwrapped steaks had greater (P<0.05) numbers of S. typhimurium than those of other treatments, whereas steaks held within the 60% CO2: 40% O2 gas atmosphere had lowest numbers overall.

Quality Changes of Beef Steaks Stored in Controlled Gas Atmospheres Containing High or Low Levels of Oxygen 1, 2.

Steaks from bovine Longissimus and Semimembranosus muscles were used to determine the influence of gas atmospheres on beef color, microbial growth and shrinkage during 9 days of retail display in two separate experimental trials. Steaks were displayed in one of four gas mixtures and were compared to steaks packaged under conventional vacuum and in a film wrap. Gas mixtures containing O2 levels of 10% (one-half ambient) did not maintain a bright red color, but those with 40-75% O2 (more than twice ambient) maintained acceptable color for 9 days of storage. Atmosphere stored steaks lost more moisture (P<0.05) than vacuum-packaged steaks. Psychrotrophic and mesophilic microbial counts from steaks stored 9 days in atmospheres containing 15% CO2 were lower (P<0.05) than the counts for the control steaks.