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ANIMAL PRODUCTS |
Department of Animal Science, Oklahoma State University, Stillwater 74078
| Abstract |
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30 d preslaughter), tissue damage, alterations in histological and collagen properties, and increased meat toughness were not observed.
Key Words: Beef Incidence Injection Lesions Tenderness
| Introduction |
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Pharmaceuticals are commonly administered to cattle at various stages of their lives (Taylor and Field, 1999
), and if given i.m., tissue damage can occur (George et al., 1995
). The National Cattlemens Beef Association has recommended that s.c. injections be administered when allowable; however, treating cattle in open-pasture situations lends to potential problems, including the stress of being held from the herd as well as unwanted restraint techniques.
Until recently, administering biological and pharmaceutical products to animals has meant that needles and syringes would be required. SolidTech Animal Health Inc., Newcastle, OK, has devised a method that uses an air-powered delivery system and biodegradable projectiles containing products such as freeze-dried ceftiofur sodium. "Biobullets" (BB) penetrate into the animals muscle and begin to be absorbed in the body, but nothing is known about the effect of this delivery system on tissue damage. Therefore, the objectives of this investigation were to determine 1) whether BB technology poses a quality control problem by creating injection site lesions, 2) what effect, if any, the BB has on any pathological changes of beef round muscle, and 3) what influence BB administration has on cooked beef tenderness.
| Materials and Methods |
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Steer calves (n = 25), of known history, located at the Willard Sparks Beef Cattle Research Facility, Oklahoma State University, were selected for use in this study. Steers had received no previous injections in any of the round muscles before the initiation of the trial and were individually identified and randomly assigned to a product administration treatment date (7, 14, 21, 28 or 35 d before slaughter).
The product administered in the trial was a standard BB casing containing 100 mg of freeze-dried ceftiofur sodium (Naxcel; Pharmacia & Upjohn, Kalamazoo, MI). At each product administration period, five steers were identified and placed into a standard commercial restraining chute. After highlighting the targeted administration area over the biceps femoris (BF), the BB was administered from a distance of 6.09 m by a trained SolidTech Animal Health representative. It should be noted that all steers received their respective BB in the identified targeted muscle location, and the average BB penetration was 3.7 cm into the targeted location. When visual evidence of BB presence was observed, tissue penetration depth was quantified using a digital electronic caliper (Brown and Sharp, North Kingstown, RI).
At the completion of the finishing period (120 d), steers were humanely slaughtered using conventional commercial procedures at the Excel beef processing facility in Dodge City, KS. After arrival at the Willard Sparks Beef Cattle Research Facility, each steer was weighed, given an individually numbered ear tag, and vaccinated with Bovishield 4+ Lepto (Pfizer Animal Health, Groton, CT). Following processing, steers were stratified by initial BW and assigned randomly within BW strata to one of five product administration treatment dates. Steers were fed a corn-based 90% concentrate diet (65.1 and 12.55 for DM and CP, respectively), which was mixed in a 1.27-m3 capacity paddle mixer. Once the total diet was mixed, the amount of feed allotted to each pen was delivered to individual pens using a computer-controlled feeding system. Two trained Oklahoma State University personnel collected carcass data, and the average score for each trait was recorded. Factors used to determine quality grade were monitored to remain consistent with the on-site USDA grading personnel. After carcass data collection, carcasses were fabricated according to institutional meat purchase specifications (IMPS; USDA, 1996
), and outside round flats (IMPS #171A) were individually identified, collected from both carcass sides, vacuum-packaged, and aged 14 d postmortem at approximately 1°C. After the aging period, each BF sample was trimmed free of subcutaneous fat and evaluated for the presence of injection site lesions. After fat removal, each BB-treated muscle section was dissected into 1.27-cm steaks (n = 15), followed by observation and palpation for the presence of injection site lesions. If any muscle tissue damage was exposed, the affected tissue was excised and weighed (to the nearest 0.3 g), along with the lesion being verbally expressed using the five-point classification system described by Dexter et al. (1994)
. Steaks were subsequently vacuum-packaged and stored at 28°C until Warner-Bratzler shear force (WBSF) determinations could be conducted.
Warner-Bratzler Shear Force
Steaks were assigned randomly to a cooking order across BB-administration time. Steaks were allowed to thaw for 24 h at 4°C before cooking. Steaks were then broiled in an impingement oven (model 1132-000-A; Lincoln Impinger, Fort Wayne, IN) at 180°C to an internal temperature of 70°C. Internal steak temperatures were monitored with copper constantan thermocouples (model OM-202; Omega Engineering, Inc., Stamford, CT.). Individual steak weights were recorded before and after cooking to determine cook loss percents. After steaks had cooled for at least 2 h to 25°C, 1.27-cm-diameter cores were removed parallel to the muscle fiber orientation. Following the procedure outlined by George et al. (1995)
, a core was removed from the immediate area near the BB administration location, and three additional cores were taken at a radial distance of 2.54, 5.08, and 7.62 cm from the administration location. Each core was sheared once by a Warner-Bratzler shear device attached to an Instron Universal Testing Machine (model 4502; Instron Corp., Canton, MA) at a crosshead speed of 200 mm/min. Peak force (kg) of cores was recorded using software provided by the Instron Corp. The average WBSF at the lesion site and the average of the WBSF for three cores at each distance of 2.54, 5.08, and 7.62 cm from the lesion location were calculated and recorded for each steak. Control samples had cores removed from the same anatomical locations as steaks obtained from the opposite side, BB-treated BF.
Histological Examination
Histopathological examination of the BB-treated and control BF muscle samples was performed by the Oklahoma Animal Disease Diagnostic Laboratory in Stillwater. Duplicate tissue samples (n = 8 control and n = 4 BB-treated) were placed in 10% formaldehyde solution for fixation and coded for submission, such that the presence/absence of a lesion, time of BB administration before slaughter, and/or the distance of the sample from the real or counterpart lesion center was unknown to the pathologist evaluating the histological sections. In all, 24 slides were prepared using Massons trichrome connective tissue stain (Luna, 1968
).
Chemical Analyses
Laboratory analyses of BF samples were conducted in duplicate according to procedures outlined by AOAC (1990
). Each sample was frozen individually in liquid N and pulverized in a Waring blender (Dynamics Co. of America, New Hartford, CT). Three grams of the powdered sample was placed in glass thimbles, dried at 100°C for 24 h, desiccated for 1 h, and reweighed to determine moisture. Following moisture determination, each sample was placed in a Soxhlet for 24 h for ether extraction of lipid, followed by drying at 100°C for no more than 12 h. Samples were then desiccated and reweighed to calculate lipid content. Using a combustion analyzer (model FP-428; Leco Corp., St. Joseph, MI), N content was determined and recorded from a separate 0.5-g pulverized sample.
Collagen Determination
To assist in explaining the differences in WBSF, collagen fractions (total, soluble, and insoluble collagen) were separated according to the procedure of Hill (1966)
. Spectrophotometric determination of hydroxyproline in the soluble and insoluble fractions was performed (Bergmann and Loxley, 1963
), and the conversion factors used for quantifying soluble and insoluble collagen were 7.52 and 7.25, respectively (Cross et al., 1973
).
Statistical Analyses
Data representing injection site lesion presence were analyzed using the frequency procedure of SAS (SAS Inst., Inc., Cary, NC). Significant differences between incidence values as associated with product administration type and time of administration were determined by calculating the
2 statistic. Means representing the lesion weights, concentrations of soluble and insoluble collagen, muscle, connective tissue, and fat proportions, and WBSF were computed, and ANOVA was determined using the GLM procedure of SAS. Each steer was used as an experimental unit within a randomized complete block design, the type of product administration and time of injection were examined as main effects, and lesion weight and WBSF value were the dependent variables. When the main effect was significant (P < 0.05), least squares means separation was accomplished by the PDIFF option of SAS (a pairwise t-test).
| Results and Discussion |
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21 d before slaughter had no (P = 0.88) detectable injection site lesions in BF. As expected, no injection site lesions were observed in the opposite side muscles (untreated controls). In the most recent national audit estimating the incidence of injection site lesions in beef top sirloin butts, Roeber et al. (2001)
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Mean WBSF measurements recorded for the BF in the current study were very similar to values reported in the National Beef Tenderness Survey1998 (Brooks et al., 2000
). Shear force values of cores from the lesion site and sites located 2.54, 5.08, and 7.62 cm away from the lesion core were 5.68, 5.34, 5.26, and 4.99 kg, respectively (Figure 1
), for steaks from BB-injected steers, whereas corresponding WBSF values from control steaks were 5.20, 5.20, 5.41, and 5.02 kg, respectively. Samples isolated from the lesion core location of BB steaks and control samples located 5.08 cm from core locations had similar (P = 0.089) WBSF values compared with remaining treated and control samples. Remaining administration times (21, 28, and 35 d before slaughter) and muscle samples locations displayed similar (P = 0.471) WBSF values. Only steers treated with the BB-implant at 7 and 14 d before slaughter displayed the presence of injection lesions in the BF; thus, no detrimental effects on beef tenderness would likely be realized with BB treatment 21 d or more before slaughter.
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In the current study, increases in total, soluble, and insoluble collagen concentrations at the lesion center decreased (P < 0.05) in concentration as the radius from the lesion center increased (Figure 2
). This result would imply that a fibroproliferative process occurred subsequent to i.m. injection of a pharmacological agent, forming a lesion core and resulting in cooked meat toughness. Sherman et al. (1980)
reported that with connective tissue reactions in wound healing, or in a fibroproliferative process, there is initially neosynthesis of collagens of pericellular type V and basement membrane type IV. Eventually, synthesis and deposition of fine, fibrillar type III collagen occurs, which is followed by the formation of matrix composed of interstitial type I collagen that resembles scar tissue. Moreover, this type I collagen is reported to have a larger fiber diameter than type III collagen, which was correlated with decreased muscle tenderness (Gay, 1983
). Concurrent with this increase in concentration of collagen and the increase in diameter of collagen fibrils, collagen solubility decreases, especially with progressing development of heat-stable covalent interchain bonds (Bailey, 1972
). It should be noted that George et al. (1995)
detected a very pronounced muscle toughening effect, as far as 7.62 cm away from the lesion core. In the current study, however, only the lesion core in 7- and 14-d samples displayed increased toughness as a result of treating cattle with a BB.
Histological examination of all samples confirmed the diagnosis of injection site lesions as described by George et al. (1995)
. From visual estimations, the relative percentages of connective tissue, muscle, and fat (Figure 3
) were 33.25, 40.23, and 26.52%, respectively, at the site of the lesion core; 11.25, 73.25, and 15.50%, respectively, at sites 2.54 cm from the lesion center; 12.25, 79.68, and 8.07%, respectively, at sites 5.08 cm from the lesion center; and 11.25, 79.98, and 9.78%, respectively, in control steaks. This finding supports results from the collagen assays, indicating that severe disruption of the relative tissue proportions had occurred in the injection lesion cores. Compared with the findings of George et al. (1995)
, which suggested that tissues as much as 5.08 cm from the lesion core were negatively altered in terms of tenderness, collagen amounts, and tissue proportions, the current findings suggest that only minor tissue alterations were evident in the BF from steers treated with the BB implant procedure immediately before slaughter.
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| Implications |
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30 d preslaughter), resulted in the absence of damage as related to unnoticeable alterations in histological and collagen properties in tissue and, in turn, led to no detectable increases in meat toughness.
1 Correspondence: 104 Animal Science (phone: 405-744-6616; fax: 405-744-7390; e-mail: bmorgan{at}okstate.edu).
Received for publication March 25, 2004. Accepted for publication July 29, 2004.
| Literature Cited |
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This article has been cited by other articles:
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M. M. Sullivan, D. L. VanOverbeke, L. A. Kinman, C. R. Krehbiel, G. G. Hilton, and J. B. Morgan Comparison of the Biobullet versus traditional pharmaceutical injection techniques on injection-site tissue damage and tenderness in beef subprimals J Anim Sci, February 1, 2009; 87(2): 716 - 722. [Abstract] [Full Text] [PDF] |
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