Relationship between wooden breast severity in broiler chicken, antioxidant enzyme activity and markers of energy metabolism.

This study aims to provide new insight on the association between the development of wooden breast myopathy and mitochondrial and glycolytic activity under oxidative stress. Myopathic muscle had higher oxidative stress together with altered glycolytic metabolism and tricarboxylic acid (TCA) cycle. This was evidenced by significantly elevated antioxidant enzyme activities (catalase, superoxide dismutase, and glutathione peroxidase), decreased citrate synthase activity and postmortem glycolytic potential with increasing wooden breast severity. In addition, affected muscles also exhibited higher initial and ultimate pH values as well as reduced total glucose and lactate contents. Citrate synthase activity was negatively correlated to antioxidant enzyme activities. Taken together, we propose that the development of the wooden breast lesion is a chronic process that may be related to the failure of muscle fibers to defend against the excessively generated oxidative products promoted by mitochondrial damage accompanied by impaired TCA cycle. Furthermore, there was a positive correlation between citrate synthase activity and glycolytic potential, which suggests that the wooden breast condition is linked to the overall altered energy metabolism of the muscle, including the oxidative phosphorylation and glycolytic pathways.

Effects of Wooden Breast Syndrome in Broiler Chicken on Sarcoplasmic, Myofibrillar, and Connective Tissue Proteins and Their Association with Muscle Fiber Area.

This study was conducted on chicken pectoralis major muscle with different wooden breast severity in combination with different sampling locations to investigate the effects of wooden breast syndrome on protein traits and total myofiber area, and their associations. Contents of sarcoplasmic, salt-soluble myofibrillar and salt-insoluble protein and proportion of total myofiber area significantly declined with increasing severity in the superficial part of muscle, whereas the amount of heat-soluble/insoluble collagen and protein denaturation as well as the area of degenerated myofibers, connective tissue and cellular infiltrates increased. Myofibril protein content indicators showed strong positive correlations to total myofiber area. Moreover, PCA results indicated that severe wooden breast is positively linked to muscle collagen content and to protein denaturation. Our results suggest that decrease in sarcoplasmic and myofibrillar proteins is associated with reduction of myofiber area. In turn, the muscle fibers are replaced by connective tissue, accompanied by excessive myofibrillar and sarcoplasmic protein denaturation.

Changes of Raw Texture, Intramuscular Connective Tissue Properties and Collagen Profiles in Broiler Wooden Breast during Early Storage.

A recently identified broiler myopathy known as wooden breast (WB) is predominantly found in the pectoralis major muscle of fast-growing broiler hybrids and is causing significant losses to the poultry industry. The aim of this study was to investigate the effects of WB syndrome on raw meat texture, purge loss and thermal properties of intramuscular connective tissue of pectoralis major muscle in the early postmortem period (1-3 days). Results showed that the presence of the WB muscles condition at 1 day postmortem was associated with significantly increased stiffness (27.0 N vs. 23.1 N) and significantly increased purge loss (1.8% vs. 1.0%) compared to normal breast (NB). However, on 3 days postmortem, these parameters did not differ between WB and NB groups. Insoluble and total collagen content was significantly higher in WB muscles compared to NB muscles, and the extractability of intramuscular connective tissue (IMCT) of WB was also higher (0.42% vs. 0.37%) compared to NB and remained stable in the early postmortem period. There was significantly lower protein content in the sarcoplasmic protein fraction and myofibrillar protein fraction of WB muscles compared to NB muscles (p < 0.05). The IMCT of these two groups showed different thermal properties, as the enthalpy of denaturation (ΔH) was significantly lower in WB muscles compared to NB muscles. The WB syndrome had a great effect on the texture and connective tissue properties of the meat compared to normal muscle, with a tendency for having a lower purge loss and higher raw meat hardness.

Hybrid Sausages Using Pork and Cricket Flour: Texture and Oxidative Storage Stability.

This study aimed to study the functionalities of cricket flour (CF) and the effects of the addition of CF on the texture and oxidative stability of hybrid sausages made from lean pork and CF. Functional properties of CF, including protein solubility, water-holding capacity, and gelling capacity, were examined at different pHs, NaCl concentrations, and CF contents in laboratory tests. The protein solubility of CF was significantly affected by pH, being at its lowest at pH 5 (within the range 2-10), and the highest protein solubility toward NaCl concentrations was found at 1.0 M (at pH 6.8). A gel was formed when the CF content was ≥10%. A control sausage was made from lean pork, pork fat, salt, phosphate, and ice water. Three different hybrid sausages were formulated by adding CF at 1%, 2.5%, and 5.0% levels on top of the base (control) recipe. In comparison to control sausage, the textural properties of the CF sausages in terms of hardness, springiness, cohesiveness, chewiness, resilience, and fracturability decreased significantly, which corresponded to the rheological results of the raw sausage batter when heated at a higher temperature range (~45-80 °C). The addition of CF to the base recipe accelerated both lipid and protein oxidation during 14 days of storage, as indicated by the changes in TBARS and carbonyls and the loss of free thiols and tryptophan fluorescence intensity. These results suggest that the addition of CF, even at low levels (≤5%), had negative effects on the texture and oxidative stability of the hybrid sausages.

Relationship between pre-rigor temperature of pork longissimus muscle, myofibril-bound calpain activity and protein degradation.

The effect of pre-rigor temperature incubation on the activity and distribution in sarcoplasmic and myofibrillar fractions of calpains, and meat quality attributes was investigated. Porcine longissimus thoracis muscles were incubated pre-rigor at 14, 22, 30 and 38 °C to 6 h postmortem, followed by another 2 h incubation at 14 °C. Thereafter, muscles were stored at 2 °C for 1 or 4 days. With higher pre-rigor temperature, sarcoplasmic Ca2+ concentration, purge loss and myofibril-bound calpain-1 content increased, while shear force declined. Water-holding capacity of isolated myofibrils was lower after pre-rigor incubation at 38 °C. Desmin and troponin T degradation, and myofibril fragmentation was greater upon incubation of isolated myofibrils with added Ca2+ in the order 800 µM Ca2+ > 40 µM Ca2+ > no Ca2+, suggesting that calpain-1 and calpain-2 were associated to myofibrils and proteolytically active with sufficient Ca2+. Activity of myofibril-bound calpain-1 in muscle incubated pre-rigor at 22 and 30 °C were higher than when incubated at 14 and 38 °C. These results indicate that calpains translocate from the sarcoplasm onto myofibrils with higher pre-rigor temperature to 30 °C and the proteolytic potential of myofibril-associated calpains is thereby increased.

Role of low molecular additives in the myofibrillar protein gelation: underlying mechanisms and recent applications.

Understanding mechanisms of myofibrillar protein gelation is important for development of gel-type muscle foods. The protein-protein interactions are largely responsible for the heat-induced gelation. Exogenous additives have been extensively applied to improve gelling properties of myofibrillar proteins. Research has been carried out to investigate effects of different additives on protein gelation, among which low molecular substances as one of the most abundant additives have been recently implicated in the modifications of intermolecular interactions. In this review, the processes of myosin dissociation under salt and the subsequent interaction via intermolecular forces are elaborated. The underlying mechanisms focusing on the role of low molecular additives in myofibrillar protein interactions during gelation particularly in relation to modifications of the intermolecular forces are comprehensively discussed, and six different additives i.e. metal ions, phosphates, amino acids, hydrolysates, phenols and edible oils are involved. The promoting effect of low molecular additives on protein interactions is highly attributed to the strengthened hydrophobic interactions providing explanations for improved gelation. Other intermolecular forces i.e. covalent bonds, ionic and hydrogen bonds could also be influenced depending on varieties of additives. This review can hopefully be used as a reference for the development of gel-type muscle foods in the future.

Role of freezing-induced myofibrillar protein denaturation in the generation of thaw loss: A review.

Formation of thaw loss cannot generally be avoided when meat is frozen and then thawed. Explanations have mainly focused on the damage to muscle fibers resulting from ice crystallization and the freezing-induced denaturation of myofibrillar proteins, the latter of which has, however, not received much research focus. This review discusses the relationship between myofibrillar protein denaturation and water-holding capacity of meat in freezing-thawing with the aim to improve the understanding the relative importance of protein denaturation in the formation of thaw loss. The contribution of decreased pH and high ionic strength in the unfrozen water in freezing is emphasized and we hypothesize that these two factors are causing protein denaturation and conformational changes within muscle fibers, and consequently loss of water-holding capacity. Slow freezing produces more thaw loss than fast freezing, and this is discussed here in relation to the impacts on myofibrillar protein denaturation induced by the freezing rate.

Myofibrillar protein characteristics of fast or slow frozen pork during subsequent storage at -3 °C.

This study aimed to investigate the effect of storage at -3 °C on myofibrillar protein in fast or slow frozen pork. Five pork loins at 48 h post-mortem were subjected to either fast (cold metal plate/-80 °C) or slow freezing (still air/-20 °C) followed by storage at -3 °C for 0, 1, 3, and 7 days before thawing. Freezing rate significantly influenced myofibrillar proteins within 3 days at -3 °C, evidenced by higher thaw loss, higher surface hydrophobicity and reduced water-holding of myofibrils, and accelerated appearance of a myosin-4 fragment (160 kDa) in slow freezing. However, these observed differences disappeared after 7 days of storage at -3 °C. The meat pH after thawing did not differ between fast and slow freezing rate. However, the pH values after thawing in both groups decreased with extended storage at -3 °C. Our results suggest that the beneficial effects of fast freezing are gradually lost by holding at -3 °C due to more extensive protein denaturation.

Mimicking myofibrillar protein denaturation in frozen-thawed meat: Effect of pH at high ionic strength.

This study aims at providing new insight on protein denaturation in freezing-thawing. Freezing-thawing minced pork reduced water-holding of myofibrils and increased surface hydrophobicity. One additional freezing-thawing cycle at slow freezing rate caused appearance of a 160 kDa myosin-4 fragment in SDS-PAGE, further decreased water-holding of myofibrils and increased surface hydrophobicity. Fresh minced pork was exposed to either high salt (2 M KCl) only or high salt with lower pH to mimic conditions in freezing. Exposure to high salt only increased water-holding of myofibrils and hence did not reproduce myofibrillar protein changes in freezing. Exposure to combinations of lower pHs and high salt decreased water-holding and increased surface hydrophobicity, suggesting myofibrillar protein denaturation occurred by a comparable mechanism as in freezing-thawing. We propose that exposure to decreased pH combined with high solute concentrations in the unfrozen water of frozen meat is the primary cause of myofibrillar protein denaturation in frozen-thawed meat.

Altered Sarcomeric Structure and Function in Woody Breast Myopathy of Avian Pectoralis Major Muscle.

The "Woody" or "Wooden" breast disease is a severe myopathy of pectoralis major muscle recently identified within rapidly growing broiler lines all around the world with a prevalence rate around 20%, or even higher. Although of significant ethical and economic impact, little is known regarding the structural and functional aspects of the contractile apparatus in the woody breast muscle. The aim of the present study was to determine physiological properties of the contractile system in the morphologically intact muscle fibers of focally damaged woody breast in comparison with normal muscle fibers to gain insight into the muscle function of the animal and possibly mechanisms involved in the disease development. Muscle samples were taken from woody breast (non-lesioned areas) and normal breast muscles from broilers. Length-tension curves, maximal active stress, maximal shortening velocity, calcium sensitivity, rate of tension development, lattice spacing and muscle biochemical composition were investigated on single skinned fibers. Sarcomeres of woody breast fibers were more compliant, which is very likely related to the wider spacing (18% wider compared to controls) between thick and thin filament. No differences were found in optimal sarcomere length (2.68 ± 0.04 vs. 2.65 ± 0.05 µm) nor in maximal active stress (116 ± 17 vs. 125 ± 19 mN mm-2). However, woody breast fibers had less steep descending arm as shown in length-tension curve. Woody breast muscle fibers had 40% bigger sarcomeric volume compared to controls. Content of contractile proteins (myosin and actin), and maximal shortening velocity were unchanged indicating that the growth in woody breast muscle fiber was associated with synthesis of new contractile units with unaltered kinetics. Calcium sensitivity was decreased in woody breast muscle fibers significantly. In conclusion, the results show that the rapid growth of muscle in woody breast disease is associated with significant structural and functional changes in the pectoralis major musculature, associated with alterations in the mechanical anchoring of contractile filaments.

Superficial and deep changes of histology, texture and particle size distribution in broiler wooden breast muscle during refrigerated storage.

Recently the poultry industry faced an emerging muscle abnormality termed wooden breast (WB), the prevalence of which has dramatically increased in the past few years. Considering the incomplete knowledge concerning this condition and the lack of information on possible variations due to the intra-fillet sampling locations (superficial vs. deep position) and aging of the samples, this study aimed at investigating the effect of 7-d storage of broiler breast muscles on histology, texture, and particle size distribution, evaluating whether the sampling position exerts a relevant role in determining the main features of WB. With regard to the histological observations, severe myodegeneration accompanied by accumulation of connective tissue was observed within the WB cases, irrespective of the intra-fillet sampling position. No changes in the histological traits took place during the aging in either the normal or the WB samples. As to textural traits, although a progressive tenderization process took place during storage (P ≤ 0.001), the differences among the groups were mainly detected when raw meat rather than cooked was analyzed, with the WB samples exhibiting the highest (P ≤ 0.001) 80% compression values. In spite of the increased amount of connective tissue components in the WB cases, their thermally labile cross-links will account for the similar compression and shear-force values as normal breast cases when measured on cooked samples. Similarly, the enlargement of extracellular matrix and fibrosis might contribute in explaining the different fragmentation patterns observed between the superficial and the deep layer in the WB samples, with the superficial part exhibiting a higher amount of larger particles and an increase in particles with larger size during storage, compared to normal breasts.

Muscle structure, sarcomere length and influences on meat quality: A review.

The basic contractile unit of muscle, the sarcomere, will contract as the muscle goes into rigor post-mortem. Depending on the conditions, such as the rate of pH decline, the cooling rate and the mechanical restraints on the muscles, this longitudinal shortening will result in various post-mortem sarcomere lengths as well as lateral differences in the distances between the myosin and actin filaments. This shortening is underlying the phenomena described as rigor contraction, thaw rigor, cold shortening and heat shortening. The shortening in combination with the molecular architecture of the sarcomere as defined by the myosin filaments and their S-1 and S-2 units, the interaction with the actin filaments, and the boundaries formed by the Z-disks will subsequently influence basic meat quality traits including tenderness and water-holding capacity. Biochemical reactions from proteolysis and glycogen metabolism interrelate with the sarcomere length in a complex manner. The sarcomere length is also influencing the eating quality of cooked meat and the water-holding in meat products.

Effect of oxygen concentration in modified atmosphere packaging on color and texture of beef patties cooked to different temperatures.

Patties were made from raw minced beef after storage for 6days in modified atmosphere (0, 20, 40, 60, and 80% oxygen) to study the combined effect of oxygen concentration and cooking temperature on hardness and color. Increased oxygen concentrations generally led to larger (P<0.01) thiobarbituric acid-reactive substances (TBARS) values, greater (P<0.01) loss of free thiols and more formation of cross-linked myosin heavy chain. Hardness of cooked patties was generally lower (P<0.01) without oxygen. Premature browning of cooked patties was observed already at a relative low oxygen concentration of 20%. The internal redness of cooked patties decreased (P<0.05) with increasing oxygen concentrations and increasing cooking temperatures. Mean particle size (D(3,2)) of homogenized cooked meat generally increased (P<0.05) with increasing cooking temperatures and increasing oxygen concentrations, and particle size was correlated (r=0.80) with hardness of cooked patties.

On the water-holding of myofibrils: Effect of sarcoplasmic protein denaturation.

The role of heat-denatured sarcoplasmic proteins in water-holding is not well understood. Here we propose a new hypothesis that in PSE-like conditions denatured sarcoplasmic proteins aggregate within and outside myofilaments, improving the water-holding of denatured myofibrils. The process is compartmentalized: 1) within the filaments the denatured sarcoplasmic proteins shrink the lattice space and water is expelled; and 2) between the myofibrils and in the extracellular space, the coagulated sarcoplasmic proteins trap the expelled water from interfilamental space. The effect of sarcoplasmic proteins on the water-holding of myofibrils following incubation for 1h at 21 to 44°C was investigated. Our results were consistent with the new hypothesis. Myofibrils without sarcoplasm had the poorest water-holding. With increasing amount of denatured sarcoplasmic proteins, the water-holding of heat-denatured myofibrils improved proportionally. X-ray diffraction was used to measure the lattice space between the filaments. Precipitated sarcoplasmic proteins shrank (P<0.001) the lattice spacing by 6.3% at 44°C.

Effect of Various Phyto-extracts on Physico-chemical, Colour, and Oxidative Stability of Pork Frankfurters.

Lipid oxidation, colour stability and physico-chemical quality of pork frankfurters with the incorporation of 0.30% sea buckthorn (SBT), 0.10% grape seed (GSE), 0.03% green tea (GTE), 0.12% fenugreek seed (FSE) and 0.10% Acacia catechu (ACE) were studied during 20 days of refrigerated aerobic storage. The SBT and ACE were identified as being the most effective antioxidants to retard lipid oxidation with the potency decreasing in the following order: SBT>ACE>GSE>GTE>FSE based on thiobarbituric acid reacting substances, peroxide value and free fatty acids. In all samples pH and aw decreased during storage period. The L* value of treated as well as control samples decreased over time while SBT and ACE exhibited an increased redness producing higher a* values than other treatments. However, GTE was more effective in increasing b* values than other treatments at the end of storage. The results suggest that functional plant-derived extracts can be valuable to the modification of frankfurter formulations for improved oxidative stability as well as quality characteristics.

Temperature induced denaturation of myosin: evidence of structural alterations of myosin subfragment-1.

Denaturation of myofibrillar proteins in porcine longissimus thoracis et lumborum muscle was investigated after pre-rigor temperature incubation at 20, 30 and 40°C. At 24h myofibrils were isolated and myosin was further cleaved by chymotrypsin. High temperature pre-rigor induced release of myosin S1 (subfragment-1), less (P < 0.05) Ca(2+)-ATPase activity and structural alterations of the region of the myosin molecule that harbors S1. Surface hydrophobicity of myofibrils from the 40°C group increased (P<0.001), suggesting a temperature-induced structural rearrangement exposing hydrophobic groups on the surface of myofibrils which in turn may explain the reduced water-holding of PSE meat.

Physiological indicators of stress and meat and carcass characteristics in tail bitten slaughter pigs.

BACKGROUND: Tail biting is a common welfare problem in pig production and in addition to being a sign of underlying welfare problems, tail biting reduces welfare in itself. The aim of this study was to evaluate the effects of tail biting on different pre and post mortem indicators of stress in slaughter pigs and on carcass and meat characteristics. A total of 12 tail bitten (TB) and 13 control (C) pigs from a farm with a long-term tail biting problem were selected for salivary cortisol analyses before and after transport to the slaughterhouse. After stunning, samples were taken for the analysis of serum cortisol, blood lactate, intestinal heat shock protein 70 (HSP70), and meat quality characteristics. In addition, body temperature immediately after and muscle temperature 35 min after stunning were measured, as well as lean meat percentage and carcass weight. RESULTS: TB pigs showed a lower cortisol response to the transport-induced stress than C pigs and also had a lower serum cortisol concentration after stunning. HSP70 content in the small intestine was higher in the TB pigs than in C pigs. TB pigs had a considerably lower carcass weight therefore produced a lower total amount of lean meat per carcass than C pigs. CONCLUSIONS: This study suggests that prolonged or repeated stress in the form of tail biting causes a blunted stress response, possibly a sign of hypocortisolism. In addition, it underlines the importance of reducing tail biting, both from an animal welfare and an economic point-of-view.

Determination of the myoglobin states in ground beef using non-invasive reflectance spectrometry and multivariate regression analysis.

Seventy-two samples of ground beef from M. semimembranosus of two 5 and two 1.5year old animals were prepared. Two types of fat tissues from either beef or pork were added to the ground beef. The samples were prepared to contain predominantly deoxymyoglobin (DMb), oxymyoglobin (OMb) and metmyoglobin (MMb) states on surfaces using selected methods based on chemical treatment (for MMb) and oxygen pressure packaging to induce the two other states. Reflectance spectra were measured on ground beef after three storage times. Partial least regression analysis was used to make calibration models of the desired myoglobin states. Validated models using leave-one-sample out cross validation gave, after correction and normalization, prediction errors of about 5%. Long term storage of ground beef was unsuitable for preparing pure MMb states due to gradual reduction of the pigment to DMb, presumably by bacteria.

Inhibition of toxigenesis of group II (nonproteolytic) Clostridium botulinum type B in meat products by using a reduced level of nitrite.

The effect of three different concentrations of sodium nitrite (0, 75, and 120 mg/kg) on growth and toxigenesis of group II (nonproteolytic) Clostridium botulinum type B was studied in Finnish wiener-type sausage, bologna-type sausage, and cooked ham. A low level of inoculum (2.0 log CFU/g) was used for wiener-type sausage and bologna-type sausage, and both low (2.0 log CFU/g) and high (4.0 log CFU/g) levels were used for cooked ham. The products were formulated and processed under simulated commercial conditions and stored at 8°C for 5 weeks. C. botulinum counts were determined in five replicate samples of each nitrite concentration at 1, 3, and 5 weeks after thermal processing. All samples were positive for C. botulinum type B. The highest C. botulinum counts were detected in nitrite-free products. Toxigenesis was observed in nitrite-free products during storage, but products containing either 75 or 120 mg/kg nitrite remained nontoxic during the 5-week study period, suggesting that spores surviving the heat treatment were unable to germinate and develop into a toxic culture in the presence of nitrite. The results suggest that the safety of processed meat products with respect to group II C. botulinum type B can be maintained even with a reduced concentration (75 mg/kg) of sodium nitrite.

Carcass and meat quality traits of four different pig crosses.

The study compared properties of Finnish Landrace×Yorkshire crossbred sows mated with Finnish Landrace (FL), Norwegian Landrace (NL), Norwegian Duroc×Norwegian Landrace (NDL), or Swedish Hampshire (SH) boars. The focus was to study the cross-sectional area of loin, cross-sectional area and number of muscle fibres, loin colour and pH value as well as the ratio of water to protein in the loin. The four studied crosses were quite similar having only small differences in carcass and meat quality. The carcass lean content was the lowest in NDL. The loins of FL and NL were longer than the loins of NDL and of SH. The cross-sectional area of loin was the largest in SH. The loin of FL was lighter in colour and the loin of SH was redder than the loins of the other crosses studied. The protein content was lower and the ratio of water to protein higher in loin of SH than in the other crosses.