Muscle gene expression associated with increased marbling in beef cattle.

The objective of this study was to identify specific bovine genes expressed within skeletal muscle that are associated with intramuscular fat deposition. Twenty-eight Angus-Simmental cross steers and heifers were harvested at the University of Illinois Meat Science Laboratory. Four pairs of animals were identified based on similar adjusted backfat thickness but differing amounts of intramuscular fat within each pair. RNA was extracted from muscle samples devoid of visible fat and microarray analysis was performed. Based on this analysis, 9 genes were selected and expression was subsequently confirmed by qPCR. Expression levels of MYH3, HOXD10, MXRA8, and CASQ2 were increased in animals with high marbling, whereas levels of NPNT, MRC1, DNER, and CYPB4 were decreased in high marbled animals. The remaining gene, ACTN2 was determined to be a false positive and was, therefore, excluded from further study. Despite the positive results of the preliminary study, associations between gene expression and intramuscular fat content did not extend to the larger population of cattle. A significant negative association existed between expression of MRC1 and marbling level (P = 0.04). Therefore, this study was unable to identify a particular skeletal muscle gene set whose expression correlated well with marbling levels in the larger population of beef cattle.

Effect of antioxidant inclusion and oil quality on broiler performance, meat quality, and lipid oxidation.

The objective of the present study was to evaluate the effect of antioxidant inclusion and oil quality on broiler performance, meat quality, shelf life, and tissue oxidative status. Ross 308 male broilers were allotted to a randomized complete block design in a 2 × 2 factorial arrangement. Factors consisted of antioxidant (ethoxyquin and propyl gallate) inclusion at 2 levels (0 or 135 mg/kg) and oil quality (fresh soybean oil, control diet peroxide value <1 mEq/kg, or oxidized soybean oil, diet peroxide value 7 mEq/kg). Each treatment included 12 pen replicates comprising 24 birds for a total of 1,152 birds on trial allotted to 48 pens. On the final day of the study, 1 bird from each pen was killed by cervical dislocation and used for determination of tissue oxidative status. Another 5 broilers from each pen were processed at a commercial slaughtering facility. Immediately after processing, carcasses were transported to the University of Illinois Meat Science Laboratory (Urbana) for further analysis. With the exception of 2 responses (liver vitamin A and serum vitamin A), no interactions were found between antioxidant inclusion and oil quality. Body weight and weight gain were increased by dietary antioxidant inclusion (P < 0.001) and fresh oil (P < 0.001). Feed intake was increased in broilers fed the antioxidant (P = 0.047) and fresh oil (P = 0.062). Antioxidant inclusion had no effect on G:F (P = 0.18). Antioxidant supplementation had no effect on carcass weight (P = 0.202), dressing percentage (P = 0.906), breast yield (P = 0.708), or breast ultimate pH (P = 0.625) and had minimal effect on breast color. Antioxidant supplementation (P = 0.057) reduced breast thiobarbituric acid reactive substances after 7 d of display. Fresh oil decreased liver thiobarbituric acid reactive substances, whereas antioxidant inclusion increased serum and liver vitamin A and E concentration. The presence of an antioxidant in the feed protects lipids from further oxidizing, therefore increasing broiler performance and improving shelf life when using oxidized oil.

Sodium citrate as a replacement for sodium chloride in a brine solution when evaluated in cows of different backfat thickness.

The objective of this study was to evaluate seven muscles from cow forequarters, which were selected based on backfat thickness; an indicator of supplemental feed before slaughter: Moderate cover (Moderate ⩾ 0.76cm) and Thin cover (Thin ⩽ 0.51cm). In addition, enhancement with a brine solution containing either Sodium Chloride or Sodium Citrate was evaluated for differences in shelf-life and meat quality. Moderate carcasses had increased carcass weight, increased fat cover and a lower yield for some of the muscles compared to Thin. However, there were minimal differences for palatability and shelf-life compared to Thin. Trained panelists detected minimal differences between Citrate and Chloride for palatability. Citrate resulted in visually darker steaks, but less discoloration during the display period. Results indicate that while selection of cow carcasses based upon backfat thickness results in minimal quality differences; compared to enhancement with Chloride, Citrate minimally impacts palatability and will extend product shelf-life by reducing discoloration.

Contribution of lean, fat, muscle color and degree of doneness to pork and beef species flavor.

Flavor is an important contributor to consumer acceptability of meat, our objective was to characterize the impact of species-specific fat/lean sources, fat level, degree of doneness and muscle color are on pork and beef flavor. Three separate experiments were conducted. Patties were formulated differently for each experiment in order to evaluate the desired variables. Experiment. 1: Flavor from combination patties (same species lean/fat or combination of species lean/fat) was not impacted by degree of doneness (66°C vs. 71°C). Beef flavor was highest in samples made with beef lean, regardless of species fat type. Pork flavor was highest in samples made with pork lean and had higher flavor intensity scores. Experiment. 2: Beef flavor was not increased in all-beef patties formulated with higher fat levels. Pork patties formulated with higher fat content increased pork flavor. Experiment. 3: All-beef and all-pork patties formulated with light or dark lean did not impact flavor in either species.

Characterization of striping in fresh, enhanced pork loins.

To investigate the striping phenomenon in fresh, enhanced pork, a series of experiments were undertaken to identify possible causes of the problem. No one factor (individual brine components, brine pH, ingredient concentration, enhancement pressure, meat and brine pH, or enhancement level) was specifically identified, which could be used to reduce the severity of the striping problem. Furthermore, tumbling the product for 2h, did not reduce the amount of striping, indicating once striping has occurred, it is permanent. Evaluation of the striping pattern indicates that the stripes are formed not only at the needle injection site, but also follow the muscle fiber orientation. The use of darker pork provided more of a contrast when evaluating striping, thus exacerbating the perceived level of striping.

The effect of pH on shelf-life of pork during aging and simulated retail display.

The pork industry uses pH to differentiate product of varying quality; thus, the effect of pH on shelf-life is important as time during transport is extended. The objective was to develop regression equations to predict shelf-life over a range of ultimate pH (5.42-6.26). Shelf-life was evaluated after vacuum aging pork loin sections 0, 7, 14, 21, or 28d and during 3d of simulated retail display (4.5°C) for pork loin chops. Correlation coefficients indicated a strong relationship between pH and quality measurements. Regression analysis with Aging Day and pH was able to explain 87% of the variation in aerobic plate counts for pork. After 28d of vacuum aging, loin sections from the upper end of the pH distribution had about a 3log(1000X) greater aerobic plate count than did the lower end pH product. An increase in pH resulted in pork with lower L*, a*, b* and R(630)-R(580) values and as Aging Day increased, instrumental measurements of color increased slightly. Although higher pH is associated with improved pork quality, higher pH and longer aging periods will result in increased microbial proliferation and decreased shelf-life. Thus, an intermediate pH may provide the most desirable combination of quality and shelf-life when extensive aging is used.

The effect of ractopamine hydrochloride (Paylean(®)) on lean carcass yields and pork quality characteristics of heavy pigs fed normal and amino acid fortified diets.

Effects of ractopamine hydrochloride (RAC) on carcass parameters in heavy weight (133.24±8.07kg) finishing pigs (n=278) given amino acid fortified (AA) or 16% crude protein (CP) diets were evaluated. A total of seven experimental diets were formulated; RAC was added at 0, 5 and 20ppm to the 16% CP diets (CP0, CP5 and CP20, respectively) and at 0, 5, 10 and 20ppm to the AA fortified diets (AA0, AA5, AA10 and AA20, respectively). Carcass, tenderloin, and ham weights were heavier (P<0.05) for RAC AA diets vs. AA0. Loin weight was heavier (P<0.05) for AA20 vs. AA0 and CP20 vs. CP0. No differences (P>0.05) were observed for color or firmness scores. Carcass muscle score, ham weight and protein% were greater (P<0.05) for RAC diets. Moisture was greater (P<0.05) and fat was lower (P<0.05) for AA5 and AA20 vs. AA0 and CP5 and CP20 vs. CP0. Feeding RAC to late finishing swine increases carcass yields and protein% with lower fat% for pigs weighing up to 136kg.

Effect of different dietary levels of natural-source vitamin E in grow-finish pigs on pork quality and shelf life.

Improving pork quality and shelf life is important in today's swine industry because higher levels of DDGS are incorporated into pig diets. Relatively high level of poly-unsaturated fatty acids (PUFA) in DDGS may increase pork susceptibility to lipid oxidation and thus reduce pork shelf life. Antioxidants such as vitamin E may delay the onset of pork lipid oxidation when used as an ingredient in the diet. This experiment examined carcass characteristics, meat quality, shelf life, and color stability in pork from pigs (n=150) fed five levels of a natural vitamin E (Nova-E) and one level of synthetic vitamin E. Natural vitamin E and synthetic vitamin E had no effect on carcass characteristics or meat quality. Increasing dietary natural vitamin E from 10 to 200mg/kg decreased lipid oxidation. Lipid oxidation of pork chops and ground pork was similar between pigs fed 40mg/kg and higher levels of natural vitamin E, indicating no additional benefits from supplementing beyond 40mg/kg natural vitamin E. Supplementing 200mg/kg synthetic vitamin E decreased pork lipid oxidation when compared to supplementing 10mg/kg natural vitamin E. High levels of natural vitamin E or synthetic vitamin E, however, did not prevent discoloration of loin chops. These data indicate that natural vitamin E was effective to help reduce lipid oxidation and the effective minimal level of dietary supplementation appeared to be 40mg/kg.

The influence of early post-mortem enhancement and accelerated chilling on pork quality.

To improve pork quality, the effectiveness of early post-mortem enhancement and accelerated chilling were investigated. The four treatments evaluated were: Enhancement with Accelerated Chilling (ENAC), Accelerated Chilling Only (ACO), Enhancement with Conventional Chilling (ENCC), and Conventional Chilling Only (CCO). ENAC had a higher (P<0.05) pH than all other treatments. CCO resulted in the highest (lightest; P<0.05) L(∗), while ENAC had the lowest L(∗) value (darkest; P<0.05). Subjective color and striping did not differ (P>0.05) between ENAC and ENCC, although ENAC was numerically higher for both parameters. Sensory analysis for juiciness and tenderness were not different (P>0.05) between ENAC and ENCC, but both were higher (P<0.05) than ACO and CCO. Enhancement early post-mortem coupled with accelerated chilling may be used to improve instrumental color and pH over conventional processing methods.

Effect of ractopamine level, gender, and duration of ractopamine on belly and bacon quality traits.

The objective was to determine belly and bacon quality traits in pigs fed ractopamine (RAC) for various durations during finishing. A 2×3×2 factorial arrangement was used with barrows and gilts, fed RAC levels of 0.0, 5.0, or 7.4ppm, for 21 or 28d prior to harvest. Bellies were fabricated and measured for length, thickness, firmness, and processing yields. Once processed, 1.27cm slices were removed at 25%, 50%, and 75% the distance from the blade end, packaged and digitally imaged using a Chem1 Genius(2) Bio Imaging System. Total slice area (TA), total slice length (TL), secondary lean length (SL), secondary lean area (SA), and percent lean area (TA - all lean components=LA) were determined by tracing images in Adobe Photoshop Elements. A composite sample from the three slices was used for proximate analysis to determine moisture and fat composition for each belly. Feeding RAC increased belly yield, TA, TL, SA, and LA (P<0.05), but did not alter moisture or fat composition (P>0.05). Gilts had decreased firmness and higher pump uptakes compared to barrows (P<0.05). Additionally gilts had increased TL, SL, and LA with lower fat and higher moisture content (P<0.05). RAC feeding duration had no significant effect on belly or bacon quality traits (P>0.05), furthermore, no interactions were found to be significant (P>0.05). RAC administration during finishing resulted in improved belly and bacon yields with no negative effects on the quality traits evaluated.

Evaluating an objective method to measure fresh pork loin firmness.

The purpose of this study was to develop an objective method for measuring fresh pork loin firmness. A total of 42 fresh boneless pork loins were collected from a range of subjective firmness scores based on the whole boneless loin to create a population suitable for creating a prediction equation. Loins were subjectively scored by panelists prior to objectively being measured with a Texture Analyzer. An R(2) value of 0.54 was achieved for the prediction equation that was generated. A second trial was performed to test the equation resulting in an R(2) value of 0.30 concerning subjective and objective values. The results from these studies indicate that it is possible to create a standardized protocol in order to determine objective firmness that would be useful for future studies and for the development of hand held or online firmness measuring devices.

Surgical anatomy of the anterior cervical spine: the disc space, vertebral artery, and associated bony structures.

OBJECTIVE: To elucidate the relationships between the neurovascular structures and surrounding bone, which are hidden from the surgeon by soft tissue, and to aid in avoiding nerve root and vertebral artery injury in anterior cervical spine surgery. METHODS: Using six cadaveric spines, we measured important landmarks on the anterior surface of the spine, the bony housing protecting the neurovascular structures in the lateral disc space, and the changes that occur during the discectomy with interbody distraction of the vertebral bodies. The measurements included the distance between the medial borders of the longus colli muscle at the level of each interspace; the width and height of each disc space at the midline; the width and height of the costal process; the distances between the cranial tip of the uncinate process (UP) and the vertebral body (VB) above and from the tip of the UP to the vertebral artery; the anteroposterior diameter or the extent of the disc spaces in the midline; the height at the midpoint of the distracted disc space; the UP-VB distance in distraction; and the width of the visible nerve root. RESULTS: The distance between the medial borders of the longus colli muscles increased in a rostral to caudal direction. The height of the UP was shortest at C4-C5 and greatest at C5-C6; the width was narrowest at C4-C5 and widest at C6-C7. The width of the costal process measured from the VB to the anterior tubercle was narrowest at C2-C3 and widest at C6-C7. The midpoint height of the costal process was smallest at C6-C7 and tallest at C4-C5 and C5-C6. The nondistracted UP-vertebral artery distance was the shortest at C2-C3 and longest at C4-C5. The nondistracted UP-VB distance averaged 1 mm at C2-C3 and C6-C7 and 1.5 mm at C4-C5. The height of the distracted disc space was shortest at C2-C3 and C6-C7. The UP-VB distance after distraction was greatest at C4-C5. Only at the C2-C3 interspace was the nerve always above the process. The vertebral artery entered the foramen transversarium of C6 in all the specimens. CONCLUSION: Although avoiding unfortunate injury is not always possible, understanding the locations and relations among the anatomic features is the only safeguard against unwarranted damage.

Development of rainbow trout microsatellite markers from repeat enriched libraries.

The efficiency of developing polymorphic microsatellite markers from 2 repeat enriched libraries was evaluated. Thirty-six polymorphic microsatellite markers were developed for rainbow trout, 27 of which were informative in a mapping family. The ability of each marker to amplify genomic DNA from other salmonids was also observed.

Effect of thyroid hormone on concentrations of plasma calcitonin in broiler chicks.

The purpose of these studies was to determine the effect of thyroidectomy (Tx), and thyroid hormone (T3/T4) treatment on concentrations of plasma CT in chicks. In addition, the turnover of CT in Tx- and T3/T4-treated chicks was estimated using a novel nonradioactive salmon CT preparation. One-week-old broiler chicks (Gallus domesticus) (n = 75) were divided into three groups. Group I was sham-injected daily (i.m. saline), Group II was injected with 50 micrograms/day of T3/T4 while Group III was injected with the goitrogen, methimazole, (150 mg/kg BW per day) for 8 weeks. Chicks (8-9 weeks old) were implanted with catheters in the brachial wing vein and administered ruthenium-labeled salmon CT. Blood samples were collected at 30 s, 1, 2, 4, 8, 20 min, and 3 h after injection. Results showed that concentrations of plasma CT were decreased in T3/T4-injected birds. There was no significant effect of methimazole on circulating concentrations of plasma CT. The half-life of CT was significantly increased (P < 0.05) in both T3/T4-injected (n = 6; 1.34 +/- 0.16 min) and goitrogen-treated birds (n = 2; 5.81 +/- 2.83 min) compared to controls (n = 7; 54 +/- 3 s) The results demonstrate that changes in concentrations of plasma thyroid hormones can significantly affect concentrations of plasma CT.

Myostatin expression and possible functions in animal muscle growth.

Myostatin (also known as growth/differentiation factor-8) is a recently identified member of the transforming growth factor-beta family of secreted regulatory factors. Mice having targeted disruption of the myostatin gene displayed a marked increase in muscle mass, up to three times normal size. Additionally, a myostatin mutation has been linked to double muscled cattle breeds characterized by a visible, generalized increase in muscle mass. Therefore, it is suggested that myostatin in muscle may be one of the long sought inhibitors that specifically control the growth of individual tissues or organs. In the present paper, we review involvement of myostatin in muscle growth of different species.

Bovine skeletal muscle calpastatin: cloning, sequence analysis, and steady-state mRNA expression.

Calpastatin is a specific inhibitor of the calpains. Calpains play a key role in postmortem tenderization of meat and have been hypothesized to be involved in muscle protein degradation in living tissue. Isolation, cloning of complementary DNA, and nucleotide sequencing of bovine calpastatin from the longissimus muscle have been completed. Two clones were identified that encompass the entire coding sequence. Clone pCR41, derived by reverse transcription-PCR, covers domains L and 1; clone pBSA1, obtained from cDNA library screening, covers domains 2 through 4 in addition to the 3'-nontranslated region. Nucleotide sequence analysis of the cDNA for bovine calpastatin revealed an average nucleotide sequence identity of approximately 70 to 80% compared with published calpastatin nucleotide sequences of human, rabbit, and pig. Exon 3, corresponding to a highly conserved 22-amino acid region, was deleted from bovine calpastatin domain L. The calculated molecular weight of bovine skeletal muscle calpastatin of 706 amino acid residues (M(r) 75,842) corresponds to the value of purified bovine skeletal muscle calpastatin as determined by SDS-PAGE (M(r) 68,000). Northern blot analysis revealed the presence of multiple calpastatin mRNA transcripts having estimated sizes of 3.8, 3.0, and 1.5 kb in beef and 3.8, 3.0, 2.5, and 1.5 kb in sheep. Calpastatin mRNA expression was increased with beta-adrenergic agonist-induced muscle hypertrophy.

A method for chronically quantifying net absorption of nutrients and gut metabolites into hepatic portal vein in conscious swine.

Surgical procedures are described for chronic cannulation of portal vein, ileal vein, abdominal aorta, and carotid artery in pigs. Silastic or Micro-Renathane tubing was used for cannulating portal vein and ileal vein, while carotid artery was cannulated with Micro-Renathane tubing. The lumen of Micro-Renathane tubing was coated with tri-dodecylmethyl ammonium chloride (TDMAC)-heparin complex. The abdominal aorta was cannulated via saphenous artery with vinyl tubing. This allows simultaneous collection of blood samples from hepatic portal vein and systemic artery (carotid or abdominal aorta) and continuous infusion of p-aminohippuric acid (PAH) into ileal vein. The constant PAH infusion provided an indicator-dilution method for estimating the blood flow rate in portal vein. In 13 pigs weighing 54 +/- 2.8 kg, the mean portal vein blood flow rate during the 8-h postprandial period was estimated to be 1,979 ml X min-1 X pig-1 or 37.8 ml X min-1 X kg-1 body weight. By simultaneously measuring the concentration of nutrients and metabolites in the portal and systemic arterial blood and multiplying porto-arterial differences by the estimated portal vein blood flow rate, the net absorption of nutrients (except long-chain fatty acids) and metabolites into hepatic portal system in conscious swine can be quantified.

Development of an enzyme-linked immunosorbent assay (ELISA) for quantification of skeletal muscle calpastatin.

An indirect antibody ELISA was developed for rapid and sensitive quantification of skeletal muscle calpastatin. Polyclonal antibodies were raised in rabbits against recombinant calpastatin, corresponding to domains 2, 3, and 4 of bovine skeletal muscle calpastatin. Western blot analysis revealed that these antibodies specifically recognize an immunoreactive calpastatin protein of approximately 130 kDa in prerigor skeletal muscle extracts. The intensity of the immunoreactive bands corresponds qualitatively with assayable calpastatin activity. For ELISA development, optimum dilutions of sample, primary anti-calpastatin antibody, and peroxidase-conjugated secondary antibody were determined by titration. A dilution optimum for coating of Immulon 4 (Dynatech) plates was observed when heated muscle extracts were diluted to 2 to 4 micrograms of protein/mL and incubated for 2 h at 37 degrees C. Optimum primary (30 micrograms IgG/mL) and secondary (Sigma A-6154; 1:1000 dilution) antibody incubations were for 1 h at 37 degrees C. Tetramethylbenzidine was used as substrate and A450 of the stopped reaction product was recorded in an automated plate reader. Calpastatin ELISA results were linearly related to calpastatin activity (calpain inhibitory activity) of heated longissimus muscle homogenates from prerigor lamb (r2 = .89; n = 40) and beef aged for 24 or 48 h (r2 = .90; n = 47). Intra-assay CV was < 5% (n = 8) and inter-assay CV was < 6% (n = 5). This assay offers advantages of speed, simplicity, and sensitivity over conventional methodology for calpastatin quantification.

Structural and metabolic integrity of sheep external intercostal muscle during extended culture.

The objectives of this study were to examine the structural and metabolic integrity of isolated sheep external intercostal muscle bundles following variable lengths of preincubation (0 to 192 h). Samples of intact external intercostal muscle (10 to 15 g), with tendons attached, were prepared from growing wethers and maintained at their resting lengths during preincubation for 0 to 192 h. Protein synthesis (PS), protein degradation (PD), acetate oxidation and ultrastructural integrity of muscle samples were examined at 0 to 192 h, 0 to 96 h, 0 to 48 h and 0 to 96 h following isolation, respectively. Additionally, the effects of variable fetal calf serum (FCS) concentrations (0 to 20%; w/v) on PS and PD and acetate oxidation were examined. Rate of PS increased as preincubation time increased to 192 h; however, most of this increase was due to the proliferation of fibroblasts on the surface of the muscle sample. Addition of cytosine arabinoside to the incubation media prevented the fibroblast-dependent increase in PS; however, it did not entirely prevent the preincubation time-dependent increase in PS. Rate of PD increased greatly upon preincubation. The nitrogen balance of incubated muscles was negative at all times examined. Acetate oxidation was maintained through 12 h of preincubation and thereafter declined. Relatively normal myofibrillar structure was maintained through 48 h of preincubation; however, loss of mitochondrial integrity and dissolution of Z-disks at 48 h and at 96 h of preincubation were evident. Isolated tissues were able to respond to FCS concentration in medium following 48 h of preincubation.

Postnatal growth of broilers in response to in ovo administration of chicken growth hormone.

The effect of in ovo administration of chicken growth hormone (cGH) on growth rate and efficiency of gain, organ, and long bone growth of 42-d-old broiler chickens was investigated. Eggs were injected once with 100 microL vehicle (0.03 M NaHCO3, 0.15 M NaCl, pH 8.3) per embryo or vehicle containing 100 ng cGH/100 microL per embryo (n = 630 eggs total) on one of the following Days: 1, 4, or 7 through 18 of embryogenesis. There was no significant difference in hatchability between control and cGH treatment groups on any given injection day. Cumulative feed conversion of all treatment groups was improved relative to their respective control groups (P < 0.05). In ovo administration of cGH on Day 15 or 16 of incubation increased body weights (P < 0.01) of female broilers. On the other hand, body weights of male broilers were significantly increased by treatment on Day 1 (P < 0.04). Breast weights of female broilers from treatment groups Day 15 or 16 were increased (P < 0.01, P < 0.05, respectively). Liver weights of female broilers from treatment groups Day 1 and 15 were increased (P < 0.05, P < 0.01, respectively). In contrast, in ovo administration of cGH on Day 11 of incubation increased liver weights of male broilers (P < 0.03). There was no significant difference between control and treatment groups, in terms of heart or leg weights, or in Warner-Bratzler shear force of Pectoralis profundus muscle. Hydroxyproline concentration and cross-sectional area of female broiler tibias from treatment groups Day 11 or Day 16 were increased (P < 0.05), and ultimate breaking strength (stress) of tibias from the same groups was reduced (P < 0.05). In ovo administration of cGH altered growth and tissue development of broiler chickens in a time by sex dependent fashion.