Effect of beef ultimate pH and large structural protein changes with aging on meat tenderness

This study investigated the effect of ultimate pH (pH_u) in beef on the degradation of large structural proteins during refrigerated storage using SDS-PAGE. M. longissimus dorsi from bull carcasses were selected and classified into three groups: low pH_u (≤ 5.79), intermediate pH_u (5.80–6.19) and high pH_u (≥ 6.2) muscles. Samples were then stored at − 1.5 °C for 1, 2, 7, 14, 21 and 28 days. Meat tenderness was measured at each aging time. Depending on meat pH_u, different protein patterns and degradation rates of structural proteins were found. Rapid changes of large structural proteins took place within 48 h post mortem. Besides titin and nebulin, degradation of filamin was clearly revealed. Two more large protein bands corresponding to myosin family members also exhibited fast decline with storage time. It suggested that the fast degradation of these proteins is a key factor in the improvement of meat tenderness.     

The effects of the ultimate pH of meat on tenderness changes during ageing

A range of ultimate pH (pH(u)) values, from 5.4 to 7.2, was produced in the m. longissimus thoracicum et lumborum (LD) of 55 sheep by subcutaneous injections of adrenaline, or by injection and subsequent exercise. The rate constant of ageing for each of 47 animals was calculated from shear force measurements, taken at intervals for up to 5 days from the LD held at 12 dgC. The relationship between the ageing rate and pH(u) could be adequately described by the equation rate constant (k) = 4.109 - 1.349(pH) + 0.1111(pH)(2); r(2) = 0.707, with a minimum rate constant at pH 6.07. The loin muscles from eight carcasses were selected to determine the effects of ultimate pH on the initial, unaged toughness value. Calpain activity was inhibited by intramuscular injection of ZnCl(2) dissolved in saline, and the muscles were aged at 10-12 °C on the carcass for up to 6 days. At all values of pH(u) (5.4-6.7), tenderness as measured by shear force were equivalent, from which it was concluded that the initial toughness of unaged lamb was not affected by pH(u). In the contralateral, untreated muscle, ageing was measured using both shear force measurement and changes in the myofibrillar fragmentation index (MFI). After 1 day, the intermediate pH range (5.8-6.2) has the highest shear force and the lowest fragmentation index value. However, in contrast to the shear force values, the ultimate MFI value in aged muscle was lowest in the high pH(u) range (6.4-6.7; minimum value at 6.4). The highest fragmentation values were found at the low pH(u) range. These observations suggest that pH(u)-related tenderness differences in meat result from different rates of ageing in the post rigor period, and therefore that these differences disappear following a sufficient ageing period. The slowest rate of tenderization measured on cooked meat was observed at pH(u) values around 6.07, but the least fragmentation of raw meat was observed at pH(u) values around 6.4, a difference of almost 0.33 pH unit.     

The influence of ante-mortem treatment on relationship between pH and tenderness of beef

Longissimus lumborum et thoracis muscles from 150 cattle comprising Czech Pied breed bulls and cows were used to investigate the relationship between pH value and beef tenderness as influenced by housing before slaughter and by the hanging mode of carcasses. The pH value was measured in the loin at the end of the slaughter-line (pH(0)) and after cooling of carcasses (pH(24),pH(48)). The texture of raw meat was measured 72h post-mortem by the Warner-Bratzler shear method; the shear force was related to pH. Curvilinear relationships with coefficients of correlation r=0.64 (P<0.05) for bulls individually housed and r=0.83 for bulls housed in groups between the shear force and the pH value 24h p.m. were found with maximal shear force (the most tough meat) in the range of pH(24) from 6.2 to 6.4. Lower correlations (r=0.72 and r=0.54) between pH(24) and meat tenderness were found in the case of cows individually housed and housed in groups, respectively. No significant relationships were found between texture and pH measured at the end of the slaughter-line (r=0.59 for bulls and r=0.65 for cows) and 48h post-mortem (r=0.64 for bulls and r=0.50 for cows). The mode of housing of cattle before slaughter influenced the properties of beef significantly. Meat of bulls housed in groups before slaughter was less tender (the shear force was higher) than meat of bulls individually housed. It could be explained by higher depletion of glycogen before slaughter and thus insufficient pH decline. Generally, the meat of cows was more tender than of bulls irrespective of housing. No significant differences were found between the left and right halves of carcasses. Thus the mode of hanging had no influence on beef tenderness.     

The effects of breed,sex, intramuscular fat and ultimate pH on pork tenderness

A study was conducted to evaluate the influence of breed and sex of animals on tenderness of pork. For evaluation, carcasses of pure breed barrows and gilts of Large White, Czech pig meat, Landrace and Duroc were used. For laboratory investigations, the samples of musculus longissimus lumborum et thoracis (20 mm thick) were taken from the same place, at the first cruciate vertebra 48 h post mortem. Seven days post mortem, the texture of heat-processed meat was measured objectively by the shear method on Instron equipped with Warner–Bratzler shear device. Meat tenderness and juiciness were also evaluated sensorially. It was observed that the tenderness of cooked meat is influenced by breed (the tenderest meat was from Duroc breed). The sex of animals has significant effect on meat tenderness too. At the same time, the effects of pH 45 minutes (pH₄₅) and 24 hours (pH₂₄) post mortem and intramuscular fat content on meat tenderness were evaluated. Correlations were found between shear force (tenderness) and intramuscular fat content and also between shear force and pH value 45 min post mortem. The shear force decreased with increasing intramuscular fat content and with increasing pH₄₅ value. Sensory evaluation of tenderness confirmed the objectively measured data.     

Influence of ultimate pH on biochemistry and quality of Longissimus lumborum steaks from Nellore bulls during ageing

Ultimate pH (pH_u) is an indicator that influences post-mortem meat quality. We studied physiological and biochemical changes of steaks obtained from Nellore bulls (Bos indicus) during post-mortem ageing. To this, Longissimus lumborum (LL) muscles were classified into three groups: Normal-pH_u (≤5.79), Intermediate-pH_u (5.80–6.29) and High-pH_u (≥6.30) groups, portioned into steaks, vacuum packaged and matured at 2 °C for 0, 7, 14, 21 and 28 days. High-pH_u steaks exhibited impaired colour stability and were darker compared to the other groups. High- and Normal-pH_u steaks showed improved tenderness and myofibrillar fragmentation linked to proteolysis. Intermediate-pH_u steaks were associated with a lower meat tenderness and decreased collagen solubility. High-pH_u steaks retained a high pH during ageing and increased water-holding capacity. These findings provide evidence that highlight pH_u as a strategy for the classification of pH_u-dependent beef quality from Nellore bulls that can be adopted by the Brazilian meat industry.     

Difference in tenderness and pH decline between water buffalo meat and beef during postmortem aging

The objective of this research was to determine the difference in tenderness and some characteristics of water buffalo meat and beef during postmortem aging. Five female crossbred water-buffalo (Philippine Carabao×Bulgarian Murrah) and five female crossbred cattle (Brahman×Philippine Native), were finished on the same diet for 6 months and slaughtered at 30 months of age. The muscle pH was measured at 40min, 3h, 7h, 24h, and 48h postmortem. Longissimus thoracis (LT) and semimembranosus (SM) muscles were excised at 2d postmortem, and shear force was measured at 2, 4, 7, and 14d postmortem. Glycogen and lactate concentrations were determined from 0, 2, and 4d LT samples, and myosin heavy chain type of buffalo and cattle LT was determined by ELISA methods. Myofibrillar protein degradation was also observed by SDS-PAGE and Western blotting of fast-type troponin T. Results showed that the buffalo meat had significantly lower shear force values compared to beef for LT and SM muscles, which was supported by a difference in troponin T degradation. Postmortem pH decline of buffalo meat was significantly slower than that of beef, which was confirmed by lactic acid concentrations, but was not explained by glycogen content. In addition, there was no significant difference in the ratio of slow to fast type muscle fibers in buffalo and cattle, indicating that myosin heavy chain type was not responsible for the difference in pH decline and tenderness between the buffalo meat and beef. This study demonstrated that the tenderness of water buffalo meat was superior to that of Brahman beef, which may have been due to the difference in pH decline and the subsequent effect on muscle protease activity.     

Effects of electrical stimulation, chilling temperature and hot-boning on the tenderness of bovine muscles

The objective of this study was to determine the effects of hot-boning, low voltage electrical stimulation (ES) and chilling temperature on the tenderness of bovine M. longissimus dorsi (LD) and M. semimembranosus (SM) muscles. LD (n=32) and SM (n=32) muscles were subjected to different post-mortem treatments; hot-boning (before 90min post-mortem), cold-boning (at 48h post-mortem), low voltage ES and rapid or slow chilling. Hot-boned muscles which were not electrically stimulated (NES) had higher Warner Bratzler shear force (WBSF) values (P<0.001) and shorter sarcomeres (P<0.001) than cold-boned muscles. Hot-boned muscles subjected to ES had lower pH values (P<0.001) post-stimulation and up to 8h post-mortem than NES muscles. At both chilling temperatures WBSF values were lower in ES hot-boned LD and SM muscles at days 2, 7 and 14 post-mortem than NES muscles. Hot-boned muscles subjected to slow chilling had longer sarcomeres (P<0.001) than those subjected to fast chilling. In hot-boned SM muscles, ES resulted in longer sarcomere lengths (P<0.001). However, ES did not have a significant effect on the sarcomere length of LD muscles. As indicated by WBSF values all muscles tenderised during ageing, including muscles which were 'cold shortened'. Proteolysis was not the main reason for differences in WBSF values between ES and NES muscles as judged by qualitative sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). A combination of slow chilling and ES had a positive effect on hot-boned muscles with respect to WBSF values.     

Effect of ultimate pH and freezing on the biochemical properties of proteins in turkey breast meat

Biochemical and functional properties of proteins from turkey breast meat with different ultimate pH at 24 h post-mortem (pH₂₄) in fresh and frozen conditions have been studied. Meat with different pH₂₄ was referred to as low, normal and high pH meat. Low and normal pH meat showed similar properties indicating their similarity in the extent of protein denaturation. Total (193 mg/g) and sarcoplasmic (99 mg/g) protein solubilities were significantly (p < 0.0001) higher in high pH meat compared to those of low and normal pH meat, and hence expected to have better functional properties. Freezing caused denaturation and oxidation of proteins as revealed by a significant decrease in Ca²⁺-ATPase activity (p < 0.0001), total protein solubility (p < 0.0001), reactive (p < 0.05) and total sulphydryl groups (p < 0.0001) and an increase in the formation of carbonyl groups (p < 0.0001), which may have implications for functionality.     

Evaluation of a probe for predicting beef tenderness

A series of three experiments were conducted to evaluate the performance of the MIRINZ (Meat Industry Research Institute of New Zealand) tenderness probe. In the first experiment, assessments on raw samples from two different muscles (longissimus lumborum and semimembranosus) were obtained using the tension head. Results indicated D50 values (torque at 50° of rotation) were the most highly related to both sensory (taste, olfactory, and feeling factors measured by a trained sensory panel) and consumer (hedonic ratings provided by untrained consumers) properties and were the values of choice, since all probe values were essentially measuring the same characteristics. In the second experiment, assessments were made using both the tension and shear heads on raw and cooked striploin (longissimus lumborum) steaks. Results confirmed probe values were essentially measuring the same characteristics, but indicated raw tension head, area 2 values (area before 50° of rotation) were more highly related to both sensory and consumer properties than were D50 values. However, probe values from raw samples did not account for a sufficient amount of variation in either sensory or consumer properties to be useful predictors of these traits, and stepwise, linear regression did not improve the predictive value of raw probe values, since all probe values were interrelated. Probe values on cooked samples indicated the shear head and area 3 values (area under the entire trace) were the most related to both sensory and consumer properties and were the values of choice. Based upon the amount of variation accounted for in sensory and consumer traits, probe area 3 values using the shear head on cooked samples provided greater predictive value for both sensory and consumer traits than Warner–Bratzler shear values obtained from steaks cooked either fresh or after being frozen and thawed. In experiment 3, correlations with individual, parametric character notes from the texture profiles of two different muscles provided no clear indication as to the textural properties being assessed by the probe. However, the MIRINZ probe appears to be a quick viable alternative to the Warner–Bratzler shear for providing an objective measure of cooked meat tenderness.     

Manipulation of the pre-rigor glycolytic behaviour of bovine M. longissimus dorsi in order to identify causes of inconsistencies in tenderness

The aim of this study was to monitor the effects of the alteration of the pre-rigor environment of the bovine LD muscle using controlled temperature regimes in order to gain an insight into the early post-mortem pH/temperature/time interactions which are important from the point of view of tenderness and to identify possible reasons for inconsistencies in beef tenderness. LD muscles (n=12) were hot-boned within 90min post-slaughter, cut into three pieces which were randomly placed in polyethylene bags and submerged in water baths pre-set at the following temperatures; 0, 5, 10, 15, 20 and 25°C for 8 h post-mortem then stored at 2°C for up to 14-days post-mortem. The rate of pH decline increased with increasing temperature. Muscles incubated at 0 and 5°C were cold shortened however not all of these muscles were tough as indicated by Warner Bratzler shear force values (WBSF). A pH range of 5.9-6.2 at 3 h post-mortem (pH(3)) produced consistently tender beef where cold-shortening was avoided. Cold shortened muscles showed the greatest variation in tenderness at 14 days post-mortem and underwent the greatest amount of tenderisation (ΔWBSF) and proteolysis between days 2 and 14 post-mortem. Proteolysis of cold shortened muscle may induce variation in tenderness in these muscles.     

Profile of biochemical traits influencing tenderness of muscles from the beef round

The objective of this study was to define the biochemical differences that govern tenderness and palatability of economically important muscles from the beef round using cuts with known tenderness differences. At 24h postmortem, the longissimus dorsi (LD), gracillus (GR), adductor (AD), semimembranosus (SM), sartorius (SAR), vastus lateralis (VL), and vastus intermedius (VI) muscles were removed from ten market weight beef cattle. Sensory and biochemical characteristics were determined in each cut and compared with the LD. The GR, SAR and VI had sensory traits similar to the LD while the SM, AD and VL differed. The GR, SAR, AD, and SM all had multiple biochemical characteristics similar to the LD, while the VI and AD had numerous biochemical differences. While no one biochemical characteristic can be used to predict tenderness across all muscles, analysis of the biochemical characteristics revealed that in most beef round cuts postmortem proteolysis provided a good indication of the tenderization occurring during aging.     

Pre rigor processing,ageing and freezing on tenderness and colour stability of lamb loins

Forty eight lamb carcasses with temperature and pH monitored were obtained from two commercial plants. At 24 h post mortem both loins (M. longissimus) from each carcass were randomly allocated to a) unaged frozen at − 18 °C, (b) aged at − 1.5 °C for 2 weeks before freezing, (c) aged for 3 weeks before freezing and (d) aged for 9 weeks without freezing. Shear force, colour stability and proteolysis were analyzed. Carcasses with a slower temperature and more rapid pH decline had more calpain autolysis, slightly higher shear force and less colour stable compared to that counterpart in general (P < 0.05). However, the shear force values of the loins were all acceptable (< 6 kgF) regardless of different pre rigor processing and ageing/freezing treatments. Furthermore, the loins aged for 2 weeks-then-frozen/thawed had a similar shear force to the loins aged only for 9 weeks suggesting that ageing-then-freezing would result in equivalent tenderness compared to aged only loins for the long-term storage.     

Influence of animal temperament and stress responsiveness on the carcass quality and beef tenderness of feedlot cattle

Three groups of steers (A, B, C) were used to study the effect of temperament (Calm, Intermediate, and Excitable) on meat quality. Temperament was based on exit velocity, pen scores, and chute scores. Temperament traits were consistent across evaluations, and values decreased (P<0.05) in magnitude over time. Increasing excitability was associated with higher (P<0.05) serum cortisol concentrations. Carcasses from cattle with calm temperaments had higher (P<0.05) 0.5h postmortem pH values than those from Intermediate and Excitable cattle (0.1 and 0.2 units, respectively). Group C Excitable steers had higher (P<0.05) WBS values than the calmer Group C steers. This trend was observed in Group A steers, although the values were not statistically different. Correlations were highest between temperament values and tenderness after 21d. Temperament influences tenderness, although the mechanism is not clear.     

Relationship between commercially available DNA analysis and phenotypic observations on beef quality and tenderness

Warner–Bratzler shear force values from 560 mixed breed heifers and steers were used to determine estimates of genetic selection. Cattle were marketed from 2008 to 2011, and included five feedlot based research projects at the North Dakota State University-Carrington Research Extension Center. Samples were collected for IGENITY® analysis providing information that included selection indices and estimated breeding values for carcass traits. DNA-based test results were compared with actual carcass measurements. Marbling accounted for over 10% of the variation in WBSF while hot carcass weight was the second most influential carcass trait accounting for 4% (P < 0.01). Regression coefficients of IGENITY® molecular breeding value on phenotype for WBSF, marbling, ribeye area, yield grade, and fat thickness were low (R² = 0.14, 0.02, 0.03, 0.03, and 0.02, respectively). Therefore selecting cattle for a higher degree of marbling and feeding a diet that meets or exceeds recommended nutrients for growth are the most important factors influencing beef tenderness and acceptability.     

Association of polymorphisms in the calpain I, calpain II and growth hormone genes with tenderness in bovine M. longissimus dorsi

The objective of this study was to determine if there is an association between tenderness in bovine M. longisimus dorsi (LD) and polymorphisms in the bovine calpain I (exons 9 & 14), calpain II (regulatory subunit) or growth hormone (intron 3) genes. Genomic DNA was isolated from bovine LD (n = 281) on which quality attributes (Warner Bratzler shear force (WBSF), sarcomere length and composition) were also characterised. DNA polymorphisms were analysed using polymerase chain reaction (PCR) and restriction fragment length polymorphism analysis. Association analyses were performed between genotypes at the four polymorphic loci and day 14 WBSF values. It was found that the calpain 1 exon 9 genotypes had an association with WBSF such that animals with the GA genotype exhibited decreased WBSF and increased tenderness when compared to animals with the GG genotype (P < 0.05). This observation concurs with that of earlier studies, suggesting that this polymorphism is a functional marker for beef tenderness.     

Near infrared spectroscopy as an on-line method to quantitatively determine glycogen and predict ultimate pH in pre rigor bovine M. longissimus dorsi

The potential of near infrared (NIR) spectroscopy as an on-line method to quantify glycogen and predict ultimate pH (pH_u) of pre rigor beef M. longissimus dorsi (LD) was assessed. NIR spectra (538 to 1677 nm) of pre rigor LD from steers, cows and bulls were collected early post mortem and measurements were made for pre rigor glycogen concentration and pH_u. Spectral and measured data were combined to develop models to quantify glycogen and predict the pH_u of pre rigor LD. NIR spectra and pre rigor predicted values obtained from quantitative models were shown to be poorly correlated against glycogen and pH_u (r² = 0.23 and 0.20, respectively). Qualitative models developed to categorise each muscle according to their pH_u were able to correctly categorise 42% of high pH_u samples. Optimum qualitative and quantitative models derived from NIR spectra found low correlation between predicted values and reference measurements.     

The intracellular distribution of small heat shock proteins in post-mortem beef is determined by ultimate pH

This study analysed Longissimus dorsi muscles from 39 Angus or Angus×Limousin bulls to determine the small heat shock protein (sHSP) dynamics in beef aged at 15°C. Using quantitative ELISA we determined that alpha β-crystallin and HSP20 were present at higher levels in muscles from Angus bulls. sHSP levels peaked at 0.5 and 3h post-mortem for HSP20 and alpha β-crystallin respectively. Intermediate pH (pH 5.7-6.3) beef had the highest level of alpha β-crystallin expression at 3-6h post-mortem. Low pH (<5.7) meat had no detected soluble alpha β-crystallin and had diminished HSP27 at 22h post-mortem. The transition of sHSP between a soluble to an insoluble phase was demonstrated by pH titration of a sarcoplasmic fraction. The effect of ultimate pH upon sHSP distribution in post-mortem muscle may influence final meat quality characteristics.     

Prediction of the time evolution of pH in meat

In this paper we compare experimental data on pH decline in carcasses and predictions using a model, based on earlier work of Vetharaniam and coworkers. This model is extended in order to cope with the varying temperatures in the slaughterhouse. We have measured initial glycogen, and the pH and temperature at multiple times in the production chain. We have obtained good comparison between model predictions and our measurements. Furthermore, the correlation between initial glycogen content and ultimate pH as predicted by the model, follows closely experimental data reported earlier in literature. Being able to predict pH decline and ultimate pH, one can obtain reliable indications of final meat quality.     

Influence of dental carcass maturity on carcass traits and meat quality of Nellore bulls

Carcasses of sixty-three Nellore bulls slaughtered at a commercial beef plant were randomly selected by dental classification (2, 4, 6 or 8 permanent incisors) in order to evaluate the influence of dental maturity on carcass traits and meat quality. Carcasses with 8 permanent incisors (p.i.) had greatest values (P<0.05) of carcass weight and longissimus area. Carcasses with 4 and 6 p.i. presented similar values of rib fat thickness being greater (P<0.05) than the other groups. Carcasses with 6 and 8 p.i. presented greater (P<0.05) values of shear force than the other groups. Conversely, carcasses with 2 and 4 p.i. displayed greater (P<0.05) myofibrillar fragmentation index and collagen solubility. Greatest values of thawing loss were observed in carcasses with 2 p.i. (P<0.05) while carcasses with 8 p.i. presented greatest values (P<0.05) of drip loss. Regarding longissimus color, carcasses with 8 p.i presented greatest value (P<0.05) of b*. Data suggests that dental maturity influences carcass traits and meat quality of Nellore bulls.