Variation in bull beef quality due to ultimate muscle pH is correlated to endopeptidase and small heat shock protein levels

This study set out to determine if ultimate pH (pH_u) affected the performance of intracellular small heat shock protein and endopeptidase dynamics in muscle during beef ageing. Longissimus dorsi muscles from 39 Angus or Limousin × Angus bulls were examined to see if pH_u achieved at 22 h post mortem (rigor) affected tenderness and water holding capacity of beef. Samples were segregated into three pH_u groups termed high (pH > 6.3), intermediate (5.7 < pH < 6.3) and low (pH < 5.7) pH_u beef. More than 30% of bull beef did not achieve acceptable tenderness at 8 days post mortem with this ageing regime. No significant differences in calpain or cathepsin enzyme levels due to meat pH were observed until after 22 h post mortem, but low pH_u beef had elevated caspase 3/7 activity soon after slaughter. At 22 h post mortem, greater levels of μ-calpain enzyme were found in the high and intermediate pH_u beef and cathepsin B levels were superior in the low pH_u beef after 2 days post mortem. Different rates of desmin and troponin T protein degradation were also observed in aged bull beef. Both proteins were degraded within 6 h post mortem for high pH_u beef, but took >3 days post mortem for intermediate pH_u beef. High levels of alpha β-crystallin (aβC) at 22 h post mortem coincided with delayed muscle protein degradation for low pH_u beef. Our results support the hypothesis that aβC shields myofibrils and buffers against endopeptidase degradation of beef structure during ageing.     

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.     

Small heat shock proteins and toughness in intermediate pHu beef

Bull M. longissimus dorsi (n = 94) categorised into high (n = 28), intermediate (n = 14) and low (n = 52) ultimate pH (pH_u) were aged at − 1.5 °C for 28 days. Shear force was higher and more variable (p < 0.05) in intermediate pH_u samples during ageing. Titin, filamin and desmin degradation was also less extensive in intermediate pH_u samples compared to the other two pH categories. The extent of the decline of HSP20, HSP27 and αβ-crystallin concentrations during post mortem ageing was pH_u related such that high pH_u meat maintained the highest concentration of small heat shock proteins followed by intermediate and low pH_u meat. μ-Calpain autolysis was slowest in intermediate pH_u and cathepsin B activities remained consistently low during ageing in this group (p < 0.05). Meat toughness in the intermediate pH_u group may be attributed to the combination of a larger pool of sHSP with a sub-optimal cathepsin B activity and intermediary μ-calpain activities.     

Influence of vacuum or high-oxygen modified atmosphere packaging on quality of beef M. longissimus dorsi steaks after different ageing times

The objective of the study was to determine if beef quality parameters differ between steaks in modified atmosphere packaging (MAP; 80% O₂ and 20% CO₂) and in vacuum packaging. Both M. longissimus dorsi (LD) from ten young bulls were cut in steaks day 3 postmortem and frozen either directly or after storage for 5 or 15 days in MAP and/or vacuum. The parameters studied were Warner Bratzler shear force, colour stability, α-tocopherol content, protein oxidation, water holding capacity and sensory attributes. Steaks stored in MAP had higher shear force than steaks stored in only vacuum at all ageing times. Tenderness and juiciness were negatively affected by storage in MAP. Ageing in MAP induced higher levels of metmyoglobin compared with vacuum ageing. In conclusion, high-oxygen MAP negatively influenced shear force, thawing loss, α-tocopherol content and colour stability, as well as the sensory attributes tenderness, juiciness and to some extent meat flavour.     

Effect of transport time on sensorial aspects of beef meat quality

Forty-eight slaughter bulls were transported by road in groups of eight for approximately 30 min, 3 h and 6 h in two replicates. After slaughter and an ageing time of 7 days, steaks from the longissimus dorsi thoracis were tasted by a trained sensory panel in terms of odour, tenderness, residue, juiciness, flavour intensity, flavour quality, and overall liking. Transport time had a significant effect (P<0.05) on meat tenderness and overall liking. The panel preferred meat from animals given the 3-h journey as opposed to the 30 min or 6-h journey and found it was more tender and had a better overall liking.     

Eating quality of “Vitela Tradicional do Montado”-PGI veal and Mertolenga-PDO veal and beef

Physicochemical and sensory characteristics were measured in veal and beef from the Portuguese Mertolenga breed having 3 quality labels as follows: Mertolenga-PDO beef and veal which apply to purebred animals and “Vitela Tradicional do Montado”-PGI veal which applies to crossbred animals. Measurements were made in longissimus lumborum muscle aged for 6 days. The temperature 3 h post-mortem (T₃), cooking losses and Warner–Bratzler shear force (WBSF) reflected carcass weight (CW) differences between groups. The pigment content was influenced by age, with beef having higher values than veal. WBSF correlated negatively with intramuscular fat in Mertolenga-PDO beef, but not on veal. WBSF correlated positively with cooking losses and negatively with myofibrillar fragmentation index, tenderness, juiciness and overall acceptability. Cooking losses and juiciness were the main contributors for the tenderness differences. Vitela Tradicional do Montado-PGI and Mertolenga-PDO veal had lighter colour and were considered tender. The three meat types were well discriminated based on pH_u, a* and C* parameters by canonical discriminant analysis.     

The effect of interaction between genotype CAST/RsaI (calpastatin) and MYOG/MspI (myogenin) on carcass and meat quality in pigs free of RYR1(T) allele

The purpose of the studies was to demonstrate to what degree genotypes of calpastatin (CAST/RsaI) and myogenin (MYOG) genes as well as the interaction between them may affect the carcass and meat quality of pigs. The investigations were conducted on 397 stress resistant pigs (free of RYR1^T allele). It was demonstrated that the favourable effect of the variants of CAST and MYOG genes on carcass quality traits depends on the cut. The gene variant favourably affecting the weight of ham simultaneously had a negative effect on the weight of the loin. It was also shown that the interaction between CAST and MYOG genotypes has a significant effect on backfat thickness. The effect of a given combination of CAST and MYOG genotypes on carcass traits is related to the weight of a substantial cut (ham, loin).Genotypes at loci CAST/RsaI and MYOG have a significant effect on the value of certain traits and parameters of meat quality and its technological value (genotype CAST on pH at 35 min and 2, 3, 24, 48, 96, 144 h post-mortem (pH₃₅, pH₂, pH₃, pH₂₄, pH₄₈, pH₉₆, pH₁₄₄, respectively), R₁ (IMP/ATP), electrical conductivity at 3 and 4 h post-mortem (EC₃, EC₄), technological yield of meat in curing and thermal processing (TY) and protein content in the muscle tissue, while genotype MYOG on pH₄₈, EC₃₅, EC₃, EC₂₄ and dry matter content).     

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.     

The protection of bovine skeletal myofibrils from proteolytic damage post mortem by small heat shock proteins

This study aimed to determine how small heat shock proteins (sHSPs) protect myofibrillar proteins from μ-calpain degradation during ageing. Immunoprecipitation experiments with M. longissimus dorsi (LD) from Angus heifers (n = 14) examined the interaction between αβ-crystallin, desmin, titin, HSP20, HSP27 and μ-calpain. Results showed that αβ-crystallin associated with desmin, titin, HSP20, HSP27 and μ-calpain. Exogenous αβ-crystallin reduced desmin and titin degradations in myofibrillar extracts and attenuated μ-calpain activity. In a second experiment, bull LD (n = 94) were aged at − 1.5 °C for up to 28 days post mortem. μ-Calpain autolysed faster in high ultimate pH (pH_u) meat (pH_u ≥ 6.2) and this was concomitant with the more rapid degradation of titin and filamin in this pH_u group. Desmin stability in intermediate pH_u meat (pH_u 5.8 to 6.19) may be due to the protection of myofibril-bound sHSPs combined with the competitive inhibition of μ-calpain by sHSPs.     

Small heat shock protein degradation could be an indicator of the extent of myofibrillar protein degradation

The objective of the present study was to evaluate the relationship between small heat shock proteins (sHSP) degradation and tenderness development of beef loins at different ultimate pH (pH_u). A total of twelve loins (M. longissimus dorsi) from steers were obtained at 1 day post mortem. Shear force and proteolysis of each loin were analyzed at 1 and 28 days post mortem. The loins at intermediate pH_u (5.8 to 6.0) showed more variation in tenderness compared to the loins at low pH_u (< 5.8), where few samples were still tough (> 10 kgF) at 28 days. The intact sHSP20 was more pronounced (P < 0.05) in the intermediate pH_u loin compared to the low pH_u counterpart. Further, high correlations between the degradation of both sHSP and myofibrillar proteins were observed (e.g. r = 0.94; degraded sHSP27 and degraded desmin). The result of this study suggests that the extent of sHSP degradation could be an indicator of myofibrillar protein degradation and tenderness.     

Relationship between rate of post mortem pH fall and ageing of Longissimus muscle in pietrain pigs

Twenty-six pietrain pigs exhibiting wide variability in pH₄₅ (pH value 45 min post mortem) values were used to assess the mechanical behaviour of cooked and raw Longissimus muscle at 24 h and 8 days post mortem. Rheological behaviour was studied in compression under strain conditions which allow specific determination of myofibrillar resistance. The muscles were divided into three groups on the basis of pH₄₅ values: (Severe pale, soft and exudative (PSE), pH₄₅ = 5.49 ± 0.03 (n = 9); PSE pH₄₅ = 5.78 ± 0.04 (n = 10); Normal, pH₄₅ = 6.35 ± 0.05 (n = 7). The three groups did not differ significantly for ultimate pH (pH_u = 5.57 ± 0.02 for the 26 muscles). A lower mechanical resistance of raw samples was found in the Normal group, regardless of the time post mortem. Although this difference was not significant, a significant relationship was found between this trait and pH₄₅ at both times post mortem (r = ?0.43, P < 0.05, at 24 h and r = ?0.44, P < 0.05, at 8 days post mortem). However, no significant effect of pH₄₅ was found on the mechanical resistance of cooked muscles. The relative decrease in myofibrillar strength after an 8-day ageing period was not affected by pH₄₅ but this was due to the fact that the three groups exhibited different starting points. Mechanical resistance of raw and cooked muscles at 24 h post mortem were significantly related to pH_u (r = ?0.55, P < 0.01, and r = ?0.49, P < 0.05, respectively). The present results show that after an 8-day ageing period, PSE muscles are not fully aged.     

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.     

Meat quality of young Nellore bulls with low and high residual feed intake

Fifty-nine Nellore bulls from low and high residual feed intake (RFI) levels were studied with the objective of evaluating meat quality traits. Animals were slaughtered when ultrasound-measured backfat thickness reached 4 mm, and samples of Longissimus were collected. A mixed model including RFI as fixed effect and herd and diet as random effects was used, and least square means were compared by t-test. More efficient animals consumed 0.730 kg dry matter/day less than less efficient animals, with similar performance. No significant differences in carcass weight, prime meat cuts proportion, chemical composition, pH, sarcomere length, or color were observed between RFI groups. Shear force, myofibrillar fragmentation index and soluble collagen content were influenced by RFI, with a higher shear force and soluble collagen content and a lower fragmentation index in low RFI animals. Feedlot-finished low RFI young Nellore bulls more efficiently convert feed into meat, presenting carcasses within quality standards.     

Effect of pre-slaughter physiological conditions on the oxidative stability of colour and lipid during chill storage of pork

The physiological condition of the live animal was found to significantly affect colour, lipid oxidation and water holding capacity of chill stored pork chops (M. Longissimus dorsi) in a study, where various pre-slaughter conditions were achieved by the following four treatments: (A) control; (B) subjected to treadmill exercise immediately prior to stunning; (C) given epinephrine injection 15 h prior to slaughter; and (D) given epinephrine injection 15 h before slaughter and further subjected to treadmill exercise immediately before stunning. The treatments resulted in variations in energy metabolites (glycogen, lactate, creatine phosphate, ATP) and ultimate pH (pH_u), with the lowest pH_u in chops from treatments A and B, and in significantly different tristimulus colour L^*-, a^*- and b^*-parameters, although the effect of treatment on colour was not consistent during the chill storage period of 6 days. Overall, chops from treatments A and B had significantly higher L^*- and b^*-values (were paler and less blue) than chops from C and D during storage under conditions typical for retail trade. The initial a^*-values were higher (redder) in chops from treatments A and B, but the colour, as judged by the a^*-values, was less stable in meat from these treatments compared with treatments C and D. Lipid oxidation, evaluated by thiobarbituric acid reactive substances (TBARS) in the fresh meat, and drip loss, measured after 6 days of storage, were both significantly higher in chops from treatments A and B compared to chops obtained from treatments C and D. Statistical analysis relating the pH and the level of various energy metabolites post-mortem in the individual animals to the measured quality parameters, revealed that pH_u was the most important factor affecting product quality. In conclusion, over all product quality depends on obtaining a pH_u in the narrow range where both meat quality parameters such as colour, lipid oxidation and drip loss as well as microbiological aspects have to be considered.     

Expression of genes related to quality of Longissimus dorsi muscle meat in Nellore (Bos indicus) and Canchim (5/8 Bos taurus × 3/8 Bos indicus) cattle

This study was performed to compare CAPN1, CAPN2, CAST, TG, DGAT1 and LEP gene expressions and correlate them with meat quality traits in two genetic groups (Nellore and Canchim) in order to assess their expression profile and use their expression profile as genetic markers. We analyzed 30 young bulls (1 year old), 15 of each genetic group. Samples of the Longissimus dorsi muscle were collected for analysis of: total lipids (TL) and meat tenderness measured as Warner-Bratzler shear force (SF) and myofibrillar fragmentation (MFI) at day of slaughter and 7 days of aging. Gene expression profiles were obtained via RT-qPCR. TL and MFI showed differences between breeds, higher MFI in Canchim and higher TL in Nellore. Calpains showed no differential expression between groups, as did DGAT1, TG, and LEP. CAST was expressed more in the Nellore cattle. The only significant within-breed correlation (0.79) between gene expression and meat traits was found for DGAT1 and MFI in Canchim breed. Although the number of animals used in this study was small, the results indicate that the increased expression of CAST in Nellore may reflect tougher meat, but the lack of correlations with the meat traits indicates it is not a promising genetic marker.     

Comparison of the thermal characteristics of connective tissue in loose structured and normal structured porcine M. semimembranosus

The aim of the present study was to characterise the thermal properties of connective tissue in loose structured porcine SM muscles in comparison to normal looking SM muscles and to see whether the meat quality traits were related to the properties of connective tissue. Samples from the muscles with loose structure and light colour were selected by visual assessment and normal looking SM muscles were used as a control (n = 25 loose structured + 25 control). The loose structured muscles had lower ultimate pH (pH_u) than the control muscles. The onset and peak temperatures of thermal shrinkage of intramuscular connective tissue (T_o and T_p, respectively) in loose structured muscles were similar to those of control muscles when the full set of data (25 loose structured + 25 control) were analysed. When the T_o and T_p data from muscles the exhibiting ten lowest and ten highest pH_u values were analysed, the low pH_u muscles (all classified as loose structured) had lower T_o and T_p than the high pH_u muscles (all classified as control) (p < 0.05). Drip loss of loose structured SM muscles (11.1%) was dramatically higher than that of control muscles (3.9%). Collagen content and collagen solubility were similar in loose structured and control muscles. It seemed that changes in the properties of intramuscular connective tissue were more easier found in porcine SM muscles with low pH_u than in SM muscles with high pH_u.     

The effects of high oxygen modified atmosphere packaging on protein oxidation of bovine M. longissimus dorsi muscle during chilled storage

Modified atmosphere packaging (MAP) extends the shelf life of beef, especially in the context of visual colour. High O₂ MAP (70-80%) may cause quality deterioration through lipid and protein oxidation, the latter linked to a reduction in meat tenderness. The objective of this study was to investigate the effects of MAP in comparison to vacuum packaging (VP) on protein oxidation and subsequent tenderness of LD (M. longissimus dorsi) beef steaks during chilled storage (4 °C) of periods up to 14 days. Steaks were analysed for carbonyl content, free thiol groups, drip loss, Warner-Bratzler shear force (WBSF) and SDS-PAGE of extracted myofibrillar proteins. Free thiol groups were lower in high O₂ MAP (80% O₂/20% CO₂) steaks, after 14 days of storage, indicating protein oxidation compared to VP steaks. SDS-PAGE (non reducing conditions) showed the presence of high molecular weight cross linked myosin heavy chain aggregates in the high O₂ atmosphere packaged steaks, which were absent in VP steaks.     

High glycolytic potential does not predict low ultimate pH in pork

Extent of postmortem pH decline influences meat quality development. To better understand physiological determination of ultimate pH (pH_u), we utilized female and castrated male pigs from a line whose selection index includes differentiated pH_u. All genotypes of AMP-activated protein kinase γ3 subunit (AMPKγ3) V199I site were present. The mutant 199II genotype increased pH_u, but only in castrated males. Genotype affected glycolytic potential (GP), but GP was weakly associated with pH_u. A subset of animals was selected based on low (− Gly) and high (+ Gly) residual glycogen content, and compared with AMPKγ3 200Q, which is associated with low pH_u. Both + Gly and 200Q muscle contained glycolytic substrate at 24 h; however, 200Q muscle generated low pH_u and greater lactate compared to + Gly. Additionally,− Gly and + Gly groups exhibited similar pH_u despite a large difference in GP. In conclusion, high GP does not appear to directly impact the extent of postmortem pH decline.     

The effect of interaction between genotype CAST/RsaI (calpastatin) and MYOG/MspI (myogenin) on carcass and meat quality in pigs free of RYR1T allele

The purpose of the studies was to demonstrate to what degree genotypes of calpastatin (CAST/RsaI) and myogenin (MYOG) genes as well as the interaction between them may affect the carcass and meat quality of pigs. The investigations were conducted on 397 stress resistant pigs (free of RYR1^T allele). It was demonstrated that the favourable effect of the variants of CAST and MYOG genes on carcass quality traits depends on the cut. The gene variant favourably affecting the weight of ham simultaneously had a negative effect on the weight of the loin. It was also shown that the interaction between CAST and MYOG genotypes has a significant effect on backfat thickness. The effect of a given combination of CAST and MYOG genotypes on carcass traits is related to the weight of a substantial cut (ham, loin).Genotypes at loci CAST/RsaI and MYOG have a significant effect on the value of certain traits and parameters of meat quality and its technological value (genotype CAST on pH at 35 min and 2, 3, 24, 48, 96, 144 h post-mortem (pH₃₅, pH₂, pH₃, pH₂₄, pH₄₈, pH₉₆, pH₁₄₄, respectively), R₁ (IMP/ATP), electrical conductivity at 3 and 4 h post-mortem (EC₃, EC₄), technological yield of meat in curing and thermal processing (TY) and protein content in the muscle tissue, while genotype MYOG on pH₄₈, EC₃₅, EC₃, EC₂₄ and dry matter content).     

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.