J. Anim. Sci. 2003. 81:669-675
© 2003 American Society of Animal Science
Effects of postexsanguination vascular infusion of carcasses with calcium chloride or a solution of saccharides, sodium chloride, and phosphates on beef display-color stability1
M. C. Hunt*,2,
J. J. Schoenbeck*,
E. J. Yancey*,
M. E. Dikeman*,
T. M. Loughin
and
P. B. Addis
* Department of Animal Sciences and Industry and
and
Department of Statistics, Kansas State University Manhattan 66506; and
and
Department of Food Science and Nutrition, University of Minnesota, St. Paul 55108
2 Correspondence:
Weber Hall (phone: 785-532-1232; fax: 785-532-7059; E-mail:
hhunt{at}oznet.ksu.edu).
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Abstract
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Hereford x Angus crossbred steers (n = 36) were stunned, exsanguinated, and infused via the carotid artery either with an aqueous solution containing 98.52% water, 0.97% saccharides, 0.23% sodium chloride, and 0.28% phosphates (MPSC; n = 12) or with 0.3 M CaCl2 (n = 12). The remaining 12 steers served as noninfused controls. At 48 h postmortem, the quadriceps muscles and subcutaneous fat were removed from the carcasses, frozen, and later made into ground beef (18 to 20% fat). The longissimus lumborum (LL), semimembranosus, and psoas major (PM) also were removed, vacuum packaged, aged until 14 d postmortem, and then one steak was sliced from each muscle for visual and instrumental color evaluations. The inside (ISM) and outside (OSM) portions of the SM were evaluated separately. The LL and OSM steaks from MPSC-infused carcasses had a lighter red (P < 0.05) initial appearance than steaks from the other treatments. The LL steaks from noninfused carcasses had the most (P < 0.05) uniform color; the MPSC treatment was intermediate, and the CaCl2 treatment was the most two-toned. Steaks from both infusion treatments had higher (P < 0.05) L* values for the LL, ISM, and OSM muscles compared with noninfused carcasses. In general, the LL from CaCl2-infused carcasses had lower (P < 0.05) a* values, saturation indices, and 630 nm to 580 nm reflectance values, and had larger (P < 0.05) hue angles. Infusion with MPSC increased (P < 0.05) hue angles in the LL and OSM. Display color stability was lowest (P < 0.05) for LL steaks from CaCl2-infused carcasses, whereas steaks from MPSC-infused carcasses were lighter red in initial color, but otherwise had display color stability similar to those from noninfused carcasses. No differences (P > 0.05) due to infusion were found for any color traits for the PM muscle and ground beef. Carotid artery vascular infusion of carcasses with CaCl2 resulted in undesirable meat colors, whereas the MPSC solution lightened loin and inside round color in a desirable way, but the color stability was slightly less compared to muscle from noninfused carcasses. Infusion effects were not consistent among muscles, and further research will be needed to determine what caused these differences.
Key Words: Beef • Carcasses • Color • Ground Beef • Infusion • pH
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Introduction
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Cardiovascular infusion of carcasses immediately after exsanguination has been reported to improve beef quality. Infusion of substrates such as CaCl2 (Koohmaraie and Shackelford, 1991) or a mixture of maltose, dextrose, glycerin, and phosphates (Farouk et al., 1992a) increased tenderness in lamb longissimus muscle. Vascular infusion (Farouk et al., 1992a) or postmortem injection (Koohmaraie et al., 1990) of substrates accelerated postmortem tenderization of the longissimus lumborum in lamb. Farouk and Price (1994) found that vascular infusion of lamb carcasses accelerated pH decline in the first 3 h postmortem. Wang et al. (1995) reported that beef carcasses infused with solutions at controlled temperatures and flow rates increased rate of chilling of the longissimus but not the semimembranosus muscle. Yancey et al. (2001) reported that vascular infusion with an aqueous solution of saccharides, sodium chloride, and phosphates plus vitamin E improved ground beef visual color scores. However, the effects of vascular infusion with either 0.3 M CaCl2 or a solution of saccharides, sodium chloride, and phosphates without added antioxidants on beef display color stability have not been addressed.
The application of infusion technology could affect meat color and stability because pumping aqueous solutions through the cardiovascular system may dilute or remove muscle pigments and create a "lighter" than normal appearance (Farouk and Price, 1994; Yancey et al., 2001). However, increased water levels (Wang et al., 1995) due to infusion may cause greater light scattering at the meat surface, thus creating a PSE appearance in the absence of true PSE conditions. The objective of our research was to determine the effects of cardiovascular infusion immediately after exsanguination with an aqueous solution of saccharides, sodium chloride, and phosphates on the initial color, uniformity of color, and display color stability of steaks.
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Materials and Methods
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Animals
Thirty-six steers (Hereford x Angus) that had been fed a typical corn-based finishing diet for 140 to 155 d, were selected visually in a commercial feedlot. Steers were shipped to the Kansas State University Beef Research Unit where they were provided feed and water until 12 h before slaughter. The steers were slaughtered in two groups of 18 (about 70 d apart). Within each group, nine steers (three animals/treatment) were slaughtered on consecutive days. The average live weight at slaughter was 537 ± 34 kg.
Slaughter and Infusion Treatments
After stunning, steers were shackled, hoisted by a rear leg, and exsanguinated through the jugular veins. Cattle were assigned randomly to treatment groups and infused to 10% of live weight via the carotid artery either with an aqueous solution containing 98.53% water, 0.97% saccharides, 0.23% sodium chloride, and 0.28% phosphates (MPSC; n = 12) or infused with 0.3 M calcium chloride (CaCl2; n = 12). The remaining 12 steers served as noninfused controls. The infusion technique was developed by MPSC, Inc. (St Paul, MN) and has been described by Yancey et al. (2001). After infusion, carcasses were dressed using normal procedures and placed in a 2°C cooler with a 1-min spray±chill cycle every 0.25 h for 8 h after cooler entry and then chilled (2°C) about 16 h without spraying.
pH and Temperature Measurements
Carcass pH decline was measured at 1, 2, 4, 8, 16, and 24 h postmortem in the triceps brachii (TB), longissimus thoracis (LT, the muscle available at ribbing), and inside semimembranosus (ISM) using a Metoxy pH electrode and meter (model HM-17MX; TOA Electronics, Ltd., Tokyo, Japan). Carcass temperature decline was monitored continuously for 24 h after cooler entry in the same three muscles using an RD-Temp-XT temperature logger with a stainless steel thermistor probe (Omega Engineering, Inc., Stamford, CT).
Fabrication
At 48-h postmortem, the longissimus lumborum (LL), psoas major (PM), semimembranosus (SM), and quadriceps (QD) muscles were excised and trimmed practically free of fat. The LL, SM, and QD muscles were vacuum packaged (in-bag pressure of 25 to 27 mm Hg) using a Multivac vacuum packager (Koch Industries, Kansas City, MO) and B-620 barrier bags (30 to 50 cc O2/m2 for 24 h at 760 torr and 23°C; Cryovac Division, W. R. Grace & Co., Duncan, SC). The PM muscles were vacuum packaged in 3-mil nylon/PE vacuum pouches (50 to 55 cc O2/m2 for 24 h at 760 torr and 21°C; Koch Industries, Kansas City, MO). The QD muscles were frozen at -40°C and the LL, PM, and SM were vacuum aged at 2°C until 14 d postmortem. Subcutaneous fat was removed from the rib and loin of each animal, vacuum packaged, and frozen (-40°C).
Ground Beef Manufacture and Steak Slicing
Fat from each carcass was thawed for 5 h at 2°C and ground (Hobart grinder, model 4732, Hobart Mfg. Co., Troy, OH) through a coarse plate (1.58 cm) and a fine plate (0.48 cm). Ground fat was stored in a -40°C freezer to keep it free flowing and minimize lipid oxidation until ground beef formulation. The QD muscles were thawed at 2°C for 48 h, removed from the vacuum package, and ground through a grinder (Biro Mfg. Co., Marblehead, OH) using a 1.27-cm plate. Frozen subcutaneous fat from each individual carcass was added to the coarse ground lean from the same carcass to achieve 18 to 20% fat by weight; it was then briefly mixed and ground twice through a 0.48-cm plate. Ground beef (454 g) from each QD muscle was prepared for display. The LL, PM, and SM muscles were removed from vacuum packages, and one steak (2.54 cm thick) was sliced from the anterior end of each for display. A second steak was sliced, frozen in liquid nitrogen, and pulverized (Waring blender) for chemical analysis. Ground beef and steaks for display were placed on foam trays (AMOCO Foam Products Corp., Atlanta, GA) with a Dri-Loc 50 meat pad (Sealed Air Corporation, Food Packaging Div., Patterson, NC.) between the meat and tray, and over wrapped with polyvinyl chloride film (21,700 cc O2/m2 at 24 h and 760 torr at 23°C; Borden, Inc., North Andover, MA). Trays were displayed in an open-topped meat display case at 1 to 3°C with two defrost cycles daily. Steaks and ground beef were illuminated with 1,614 lx of Deluxe Warm White fluorescent lighting (Phillips Lighting Co., Salina, KS).
Ground beef and steaks were evaluated by a six-member, trained color panel using scales and procedures in meat color evaluation guidelines (AMSA, 1991). Ground beef was evaluated for initial color on d 0 and display color through 4 d of display. Steaks were evaluated for initial color and color uniformity on d 0, and display color stability was scored through 4 d for the PM and through 5 d for the LL and SM muscles. Initial color (1 = pale red or bleached red, 2 = very light cherry red, 3 = moderately light cherry red, 4 = cherry red, 5 = slightly dark red, 6 = moderately dark red, 7 = dark red, and 8 = very dark red), color uniformity (1 = uniform, 2 = slight two-toning, 3 = small amount of two-toning, 4 = moderate amount of two-toning, and 5 = extreme two-toning), and display color stability (1 = very bright cherry red or pale red, 2 = bright cherry red or pale red, 3 = slightly dark red to tan or brown, 4 = moderately dark red to tan or brown, and 5 = dark red to tan or brown) were scored by panelists to the nearest 0.5 point. The SM typically has a darker red outside portion (OSM) and a lighter red inside portion (ISM), and these two muscle areas were scored separately. The outer one-third of the muscle was considered the OSM, the inner one-third was designated the ISM (middle third was not scored).
Instrumental color was evaluated throughout the display period using a Labscan 2000 (HunterLab, Inc., Reston, VA) to measure CIE L*, a*, and b* values and spectral reflectance (400 to 700 nm) using Illuminant C and a 10° observer with an aperture size of 2.54 cm. Steak and ground beef light-exposed surfaces were measured daily at three locations and the values were averaged. The hue angle (HA = tan-1 b*/a*), and saturation index (SI = [a*2+b*2][1/2]) were calculated. The difference of 630 to 580 nm, a measure of redness, was calculated from spectral data.
Statistical Analyses
The design structure was a split-split-plot where infusion treatment (n = 3) was the whole plot, muscle (n = 5) the subplot, and display time (n = 5 or 6, repeated measure) the sub-subplot (using a compound symmetry covariance structure). Within the whole plot (36 animals), each infusion treatment was assigned to three animals as a generalized randomized complete block where each slaughter group (18 animals/group slaughtered about 70 d apart) served as a block. Within each block (18 animals/group), cattle were slaughtered on two consecutive days (9 animals/d). Thus, on each slaughter day, within a group, three animals received an infusion treatment, resulting in four experimental units (three animals/unit) for each whole-plot treatment. Treatment was a fixed effect and slaughter group and slaughter day were random effects in the whole plot; treatment and muscle were fixed effects in the subplot; and treatment, muscle, and display time were fixed effects in the sub-subplot. Within the sub-subplot, a package of ground beef or steak was the subject for display time repeated measures, and visual panelist was a random effect. Data were analyzed using the PROC MIXED procedure of SAS (SAS Inst., Inc., Cary, NC), and interactions were analyzed with the highest order interactions (P < 0.05) considered. Least squares means from the LSMEANS statement were separated (P < 0.05) by the DIFF option for protected (P < 0.05) F-tests.
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Results and Discussion
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Carcass pH Decline
A greater pH decline (Table1
) occurred the first 3 to 4 h postmortem in three muscles from the infused carcasses compared to those from noninfused carcasses. It took until 4 h postmortem for the pH of the TB and ISM muscles, and 16 h for the LL of control carcasses, to equal the pH of corresponding muscles in infused carcasses. All muscles had the same (P > 0.05) pH at 24 h postmortem. Rate of temperature decline in the TB, LL, and ISM muscles were not different between muscles of infused and control carcasses (Dikeman et al., 2003). However, the lower pH values combined with higher carcass temperatures (i.e., 1 to 4 h postmortem) in both infusion treatments resulted in conditions favorable for protein denaturation, which could result in a lighter and softer muscle. Unruh et al. (1986) reported that the longissimus of electrically stimulated cattle (more rapid pH decline) had a lighter red lean color at 24 and 48 h postmortem and during the first few days of display compared to nonelectrically stimulated controls. Similar results were reported for the SM muscle (Sammel et al., 2002). Furthermore, Dikeman et al. (2003) indicated that the saccharides provided by the MPSC solution could have served as an additional energy source for anaerobic metabolism and could have contributed to the more rapid pH decline as the saccharides were utilized and lactic acid accumulated in the muscles.
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Table 1. pH decline means by treatment and muscle from carcasses which were cardiovascularlyinfused immediately after exsanguination
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Muscle pH
There were no differences in 48-h pH due to treatment for the LL, ISM, and OSM muscles. The PM from MPSC-infused carcasses had a higher (P < 0.05) pH (5.89) at 48 h than the PM from noninfused carcasses (5.78), whereas the pH of PM steaks from carcasses that were CaCl2 infused (5.83) was intermediate and not different (P > 0.05) from either the MPSC or control treatments.
Initial Color Scores
The OSM and LL muscles from the MPSC carcasses had lighter cherry-red initial color scores (P < 0.05) than steaks from the CaCl2-infused or noninfused carcasses (Table 2). Farouk and Price (1994) suggested that the lighter color of muscles from infused lamb carcasses might have resulted from increased light scattering or muscle pigment dilution. The lighter color did not appear to be a dilution of muscle pigments because Schoenbeck (1998) found that the concentrations of total pigment (myoglobin plus hemoglobin) for the LL, PM, ISM, and OSM muscles were not different between infused and noninfused beef carcasses; however, the PM muscle from CaCl2-infused carcasses retained more hemoglobin than the PM from noninfused or MPSC-infused carcasses. Thus, the differences in initial score appeared to be the result of increased light scattering associated with the added water during infusion and/or the rapid pH decline.
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Table 2. Least squares means for initial color score, color uniformity score, visual display score, a*, saturation index (SI), hue angle (HA), and reflectance at 630 to 580 nm for steaks and ground beef from carcasses cardiovascularlyinfused immediately after exsanguination
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A treatment x muscle interaction (P < 0.05) occurred for color uniformity scores. All carcass treatments in the LL muscle (Table 2
) were different (P < 0.05) from each other with uniformity from greatest to least for noninfused > MPSC infused > CaCl2 infused. The LL from noninfused carcasses essentially had a uniform color, whereas both the MPSC-infused (1.8) and CaCl2-infused (2.2) treatments had more two-toning. The CaCl2-infused treatment created a speckled or mottled brownish-red appearance that would not be acceptable for meat purveyors or consumers. In addition, both of the infusion treatments resulted in darker-red areas around the outer perimeter of the muscle and lighter-red color in the center and ventral areas of the LL muscle. These color variations may be related to the muscle contraction visible during the infusion processing. The CaCl2 caused more severe contraction than the MPSC solution, and a rigor-like state developed within 30 min of death that persisted through fabrication at 48 h postmortem (Dikeman et al., 2003). The depolarization of prerigor muscle and the Ca-induced muscle contraction were not surprising, but the extent of the contraction and associated muscle discoloration was.
There were treatment x muscle interactions (P < 0.05) for a*, saturation index (SI), hue angle (HA), and 630 to 580 nm (Table 2). With the exception of HA, these differences were found only in the LL muscle. Longissimus steaks from the noninfused control carcasses had the highest (P < 0.05) a* values (most red), those from MPSC-infused carcasses were intermediate, and steaks from the CaCl2±infused treatment were least red. Both the MPSC-infused and noninfused carcasses had a more vivid (higher SI; P < 0.05), redder color (greater 630 to 580 nm values; P < 0.05) in the LL than the CaCl2-infused carcasses (Table 2). Renerre (1990) reported that larger 630 to 580 nm values were a result of brighter red meat color, with the lower level of acceptable meat color corresponding to a value of <12.5. None of the treatments in our study was near this threshold, but the data clearly indicate that MPSC and noninfused carcasses resulted in muscles that had a lighter-red initial color than steaks from CaCl2-infused carcasses.
Display Color Stability
Three-way (treatment x muscle x display day) interactions (P < 0.05) were noted for visual display color stability scores, L*, and b* values of the LL, ISM, and OSM muscles (Table 3), but not for the PM and ground beef (data not shown). In the LL muscle, both the MPSC-infused and noninfused controls had lighter-red (P < 0.05) display scores than the CaCl2 treatment on all display days except d 0. On d 0, steaks from the MPSC treatment had the lightest-red (P < 0.05) appearance. In the ISM (d 3) and OSM (d 0 and d 1), the MPSC treatment had lighter-red (P < 0.05) visual scores. In the LL muscle, the MPSC treatment had higher (P < 0.05) L* values than the noninfused muscles on each display day. This same trend occurred in the ISM muscle with the MPSC treatment having higher L* (P < 0.05) values than the noninfused muscles on d 1, 2, and 5. In the OSM, the MPSC treatment had a greater (P < 0.05) L* value than the other two treatments on all display days except d 0. In all muscles, the MPSC treatment had the highest (P < 0.05) L* value (lightest color). Differences (P < 0.05) were found for b* on each display day for the LL muscle, on d 1 only for the ISM, and on d 1 to 4 in the OSM. For b* values, steaks from the MPSC treatment had the highest (P < 0.05) values in all muscles, which indicated a more yellow appearance when compared to the noninfused and CaCl2 carcasses.
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Table 3. Treatment x muscle x display day interaction least squares means for display color stability, L*, and b* for the longissimus lumborum (LL), inside semimembranosus (ISM), and outside semimembanosus (OSM) muscles from carcasses cardiovascularly infused immediately after exsanguination
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The obvious pattern was for display color scores to increase (more discoloration) as time progressed (Table 3). For the LL, the CaCl2-infused and noninfused treatments were not different for visual scores (P > 0.05) at d 0, but at d 1 and throughout the remaining display period, the CaCl2-infused treatment resulted in more discoloration than did the MPSC and noninfused treatments. Apparently the CaCl2 infusion caused a faster conversion of oxymyoglobin to metmyoglobin, which was perceptible to color panelists. Treatment differences were not as pronounced in the ISM and OSM muscles for visual display color stability, but these muscles tended to follow the differences found in the LL. No differences were found for color traits of the PM and ground beef (data not shown).
A significant treatment x display day interaction was found for hue angle (Figure 1) and 630 to 580 nm values (Figure 2). The CaCl2 and MPSC carcass treatments resulted in greater (P < 0.05) hue angles on d 2 to 5 of display when compared to noninfused controls. Because greater hue angles during display indicate greater discoloration towards metmyoglobin, the infused treatments had less color stability compared to the noninfused controls. On display d 1 to 5, the CaCl2 treatment resulted in lower (P < 0.05) 630 to 580 nm values than the noninfused and MPSC-infused muscles. Clearly, the steaks from CaCl2 treatment had inferior color during display.
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Figure 1. Treatment x display day interaction least squares means for hue angle values obtained from steaks and ground beef from cattle infused with either a solution of calcium chloride or a solution of saccharides, sodium chloride, and phosphates, or noninfused control cattle. Means in a day with different superscripts differ (P < 0.05).
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Figure 2. Treatment x display day interaction least squares means for reflectance values at 630 nm±580 nm obtained from steaks and ground beef from cattle infused with either a solution of calcium chloride or a solution of saccharides, sodium chloride, and phosphates, or noninfused control cattle. Means in a day with different superscripts differ (P < 0.05).
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The majority of infusion treatment-induced differences (P < 0.05) for visual and instrumental measures of beef color were noted in the LL, ISM, and OSM muscles. Clearly, carotid artery-infused solutions reached the LL, resulting in consistent treatment effects on L*, b*, and visual color stability scores. Pumping aqueous solutions to areas near the infusion site is more easily achieved than pumping infusates to muscles located in posterior portions of the carcass (Wang et al., 1995). However, in our study, color differences associated with infusion treatment were found in the ISM and OSM; thus, it is evident that cardiovascular infusion did deliver substrates to posterior portions of the carcasses. Color differences were small for the PM muscle located near the LL. The faster pH decline postmortem due to infusion was more related to lightening of muscle color than was muscle location in the carcass. Sammel et al. (2002) reported a lighter-red color and softer lean for the ISM vs OSM from carcasses known to differ in rate of chill and pH decline. Farouk and Price (1994) reported that pH declines (longissimus and infraspinatus) in the first 3 h postmortem were accelerated in lamb carcasses that were cardiovascularly infused. In our study, in the absence of infusion, the ISM was paler, presumably due to slower chilling in the deep portion of the muscle compared to the OSM. The L* values for the ISM were greater than the OSM for infused than noninfused carcasses. Thus, the lighter colored lean caused by the infused treatments was confirmed by higher L* values and lighter initial color, and by lower display color scores.
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Implications
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This novel carcass infusion technique impacted meat color stability because some of the infused substances accelerated postmortem glycolysis. Infusion of CaCl2 significantly darkened meat color, created a mottled appearance, and caused a rigor-like state that would be problematic for the meat and food industries. The lighter-red initial color of meat from carcasses infused with saccharides, salt, and phosphates would be desirable, but the slightly faster muscle discoloration would be undesirable. In addition, effects of this infusion technology will not be uniform across all muscles. More research is needed to refine this methodology such that color traits of economically important muscles would be positively affected.
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Footnotes
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1 Appreciation is expressed to the National Cattlemen’s Beef Association, Greenwood Village, CO; North American Meat Processors Association, Reston, VA; MPSC, Inc., St. Paul, MN; Koch Industries Feedlot, Wichita, KS, and the Minnesota Agric. Exp. Stn. for their financial support of this research. Contribution no. 02-229-J from the KS Agric. Exp. Stn. 
Received for publication April 19, 2002.
Accepted for publication October 19, 2002.
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References
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AMSA. 1991. Guidelines for Meat Color Evaluation. The American Livestock and Meat Board, Chicago, IL.
Dikeman, M. E., M. C. Hunt, J. Schoenbeck, P. B. Addis, E. Katsanidis, M. Pullen, and E. J. Yancey. 2003. Effects of postexsanguination vascular infusion of cattle with a solution of saccharides, sodium chloride, and phosphates or with calcium chloride on meat quality and sensory traits of steaks and ground beef. J. Anim. Sci. 81:156–166.[Abstract/Free Full Text]
Farouk, M. M., and J. F. Price. 1994. The effect of post-exsanguination infusion on the composition, exudation, color, and post-mortem metabolic changes in lamb. Meat Sci. 38:477–496.
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Schoenbeck, J. J. 1998. Effects of cardiovascular infusion on beef carcass appearance and the color and chemical traits of five muscles. M.S. Thesis, Kansas State Univ., Manhattan.
Unruh, J. A., C. L. Kastner, D. H. Kropf, M. E. Dikeman, and M. C. Hunt. 1986. Effects of low-voltage electrical stimulation on meat quality and display color stability. Meat Sci. 18:281–293.
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Abstract
Introduction
