LC-MS-based metabolomics reveals metabolite dynamic changes of beef after superchilling early post-mortem.

To explore the underlying mechanisms by which superchilling (SC, -3 °C within 5 h of slaughter) improves beef tenderness, an untargeted metabolomics strategy was employed. M. Longissimus lumborum (LL) muscles from twelve beef carcasses were assigned to either SC or very fast chilling (VFC, 0 °C within 5 h of slaughter) treatments, with conventional chilling (CC, 0 âˆ¼ 4 °C until 24 h post-mortem) serving as the control (6 per group). Biochemical properties and metabolites were investigated during the early post-mortem period. The results showed that the degradation of µ-calpain and caspase 3 occurred earlier in SC treated sample, which might be attributed to the accelerated accumulation of free Ca2+. The metabolomic profiles of samples from the SC and CC treatments were clearly distinguished based on partial least squares-discriminant analysis (PLS-DA) at each time point. It is noteworthy that more IMP and 4-hydroxyproline were found in the comparison between SC and CC treatments. According to the results of metabolic pathways analysis and the correlation analysis between traits related to tenderness and metabolites with significant differences (SC vs. CC), it can be suggested that the tenderization effect of the SC treatment may be related to the alteration of arginine and proline metabolism, and purine metabolism in the early post-mortem phase.

Secondary Lipid Oxidation Products as Modulators of Calpain-2 Functionality In Vitro.

The objective was to understand the impacts of secondary lipid oxidation products on calpain-2 activity and autolysis and, subsequently, to determine the quantity and localization of modification sites. 2-Hexenal and 4-hydroxynonenal incubation significantly decreased calpain-2 activity and slowed the progression of autolysis, while malondialdehyde had minimal impact on calpain-2 activity and autolysis. Specific modification sites were determined with LC-MS/MS, including distinct malondialdehyde modification sites on the calpain-2 catalytic and regulatory subunits. 2-Hexenal modification sites were observed on the calpain-2 catalytic subunit. Intact protein mass analysis with MALDI-MS revealed that a significant number of modifications on the calpain-2 catalytic and regulatory subunits are likely to exist. These observations confirm that specific lipid oxidation products modify calpain-2 and may affect the calpain-2 functionality. The results of these novel experiments have implications for healthy tissue metabolism, skeletal muscle growth, and post-mortem meat tenderness development.

Divergent Regulatory Roles of Transcriptional Variants of the Chicken LDB3 Gene in Muscle Shaping.

LIM domain binding 3 (LDB3) serves as a striated muscle-specific Z-band alternatively spliced protein that plays an important role in mammalian skeletal muscle development, but its regulatory role and molecular mechanism in avian muscle development are still unclear. In this study, we reanalyzed RNA sequencing data sets of 1415 samples from 21 chicken tissues published in the NCBI GEO database. First, three variants (LDB3-X, LDB3-XN1, and LDB3-XN2) generated by alternative splicing of the LDB3 gene were identified in chicken skeletal muscle, among which LDB3-XN1 and LDB3-XN2 are novel variants. LDB3-X and LDB3-XN1 are derived from exon skipping in chicken skeletal muscle at the E18-D7 stage and share three LIM domains, but LDB3-XN2 lacks a LIM domain. Our results preliminarily suggest that the formation of three variants of LDB3 is regulated by RBM20. The three splice isomers have divergent functions in skeletal muscle according to in vitro and in vivo assays. Finally, we identified the mechanism by which different variants play different roles through interactions with IGF2BP1 and MYHC, which promote the proliferation and differentiation of chicken myoblasts, in turn regulating chicken myogenesis. In conclusion, this study revealed the divergent roles of three LDB3 variants in chicken myogenesis and muscle remodeling and demonstrated their regulatory mechanism through protein-protein interactions.

A Solvent-Templated Porous Liquid Crystal Elastomer with Tactile Sensation beyond Reversible Actuation toward Versatile Artificial Muscles.

Liquid crystal elastomers (LCEs), as a classical two-way shape-memory material, are good candidates for developing artificial muscles that mimic the contraction, expansion, or rotational behavior of natural muscles. However, biomimicry is currently focused more on the actuation functions of natural muscles dominated by muscle fibers, whereas the tactile sensing functions that are dominated by neuronal receptors and synapses have not been well captured. Very few studies have reported the sensing concept for LCEs, but the signals were still donated by macroscopic actuation, that is, variations in angle or length. Herein, we develop a conductive porous LCE (CPLCE) using a solvent (dimethyl sulfoxide (DMSO))-templated photo-cross-linking strategy, followed by carbon nanotube (CNT) incorporation. The CPLCE has excellent reversible contraction/elongation behavior in a manner similar to the actuation functions of skeletal muscles. Moreover, the CPLCE shows excellent pressure-sensing performance by providing real-time electrical signals and is capable of microtouch sensing, which is very similar to natural tactile sensing. Furthermore, macroscopic actuation and tactile sensation can be integrated into a single system. Proof-of-concept studies reveal that the CPLCE-based artificial muscle is sensitive to external touch while maintaining its excellent actuation performance. The CPLCE with tactile sensation beyond reversible actuation is expected to benefit the development of versatile artificial muscles and intelligent robots.

Identifying the Potential Apoptotic Metabolites in Postmortem Beef Muscle by Targeted Metabolomics.

Apoptotic cells may release specific metabolites to act as messengers during the apoptotic process. This study represents the first attempt to identify potential apoptotic metabolites in postmortem muscle. Ninety potential apoptotic metabolites in beef were selected and analyzed through targeted metabolomics, with 84 of them exhibiting significant differences over the postmortem time. Following the addition of the mitochondria-targeted antiapoptotic agent mitoquinone to postmortem muscle, metabolomic analysis revealed that 73 apoptotic metabolites still underwent significant changes, even against the backdrop of altered apoptosis. Of these 73 apoptotic metabolites, 54 exhibited similar trends at various treatment times with adding mitoquinone, including lipids (6), amino acids (27), nucleosides (11), and carbohydrate and energy metabolism (10). Mitoquinone significantly reduced the levels of most apoptotic metabolites, and inhibition of apoptosis resulted in a significant decrease in the levels of numerous apoptotic metabolites. Consequently, these apoptotic metabolites are considered complementary to apoptosis in postmortem muscle, with their increased levels potentially promoting apoptosis. Noteworthy apoptotic metabolites, such as glycerol 3-phosphate, serine, AMP, ATP, GMP, and creatine, were identified as active signaling molecules that attract and recruit phagocytes during apoptosis, assisting in recognizing apoptotic cells by phagocytes. This study provides, for the first time, insights into potential apoptotic metabolites in postmortem muscle, contributing to a better understanding of meat biochemistry.

Validation of a handheld near-infrared spectrophotometer for measurement of chemical intramuscular fat in Australian lamb.

The objective of the study was to independently validate a calibrated commercial handheld near infrared (NIR) spectroscopic device and test its repeatability over time using phenotypically diverse populations of Australian lamb. Validation testing in eight separate data sub-groups (n = 1591 carcasses overall) demonstrated that the NIR device had moderate precision (R2 = 0.4-0.64, RMSEP = 0.70-1.22%) but fluctuated in accuracy between experimental site demonstrated by variable slopes (0.50-0.94) and biases (-0.86-0.02). The repeatability experiment (n = 10 carcasses) showed that time to scan post quartering affected NIR measurement from 0 to 24 h (P < 0.001). On average, NIR IMF% was 0.97% lower (P < 0.001) at 24 h (4.01% ± 0.166), compared to 0 h. There was no difference (P > 0.05) between Time 0 and 1 h or Time 0 and 4 h or between replicate scans within each time point. This study demonstrated the SOMA NIR device could predict lamb chemical IMF% with moderate precision and accuracy, however additional work is required to understand how loin preparation, blooming and surface hydration affect NIR measurement.

Assessing the impact of ultrasound on the rate and extent of early post-mortem glycolysis in bovine Longissimus thoracis et lumborum.

The rate of pH decline, early post-mortem, has been identified as a key factor that impacts the tenderness of meat, and manipulating this rate of pH decline is highly relevant to ensure consistent high quality meat. Ultrasound is a potential intervention in early post - mortem muscle that may have an impact on the rate of glycolysis through its ability to alter enzyme activity. Following a variety of different ultrasound treatments frequencies (25 and 45 kHz) and durations (15, 30 and 45 min), it was found, when analysed in muscle, that ultrasound treatment duration, specifically the 30 min treatment, and interaction between treatment duration and frequency, had a significant impact on the rate of pH decline, post - treatment. Frequency did not have a significant effect on the rate of pH decline, post - treatment, in muscle. Ultrasound did not have a significant permanent effect on the activity of glycolytic enzymes present in bovine Longissimus lumborum et thoracis muscle, where no significant differences were observed on the rate of pH decline and rate of change of reducing sugars, glycogen and lactic acid, when analysed in an in vitro glycolytic buffer. It seems that the impact observed in intact muscle is not as a consequence of a permanent change in enzymatic activity, instead indicating an impact on conditions in the muscle which enhanced enzyme activity.

Beef muscle discrimination based on two-trace two-dimensional correlation spectroscopy (2T2D COS) combined with snapshot visible-near infrared multispectral imaging.

The purpose of this work was to assess the potential of 2T2D COS PLS-DA (two-trace two-dimensional correlation spectroscopy and partial least squares discriminant analysis) in conjunction with Visible Near infrared multispectral imaging (MSI) as a quick, non-destructive, and precise technique for classifying three beef muscles -Longissimus thoracis, Semimembranosus, and Biceps femoris- obtained from three breeds - the Blonde d'Aquitaine, Limousine, and Aberdeen Angus. The experiment was performed on 240 muscle samples. Before performing PLS-DA, spectra were extracted from MSI images and processed by SNV (Standard Normal Variate), MSC (Multivariate Scattering Correction) or AREA (area under curve equal 1) and converted in synchronous and asynchronous 2T2D COS maps. The results of the study highlighted that combining synchronous and asynchronous 2T2D COS maps before performing PLS-DA was the best strategy to discriminate between the three muscles (100% of classification accuracy and 0% of error).

Effect of residual feed intake on meat quality in fattening Charolais bulls fed two contrasting diets.

The selection of more efficient animals for breeding is of both economic and environmental interest to the industry. The aim of this study was to evaluate the influence of the animals' residual feed intake (RFI) ranking in interaction with the type of diet on the meat quality of Charolais beef cattle. Indeed, several biological mechanisms are associated with RFI, especially when animals are fed high starch-diets. It is therefore possible that quality parameters may show greater changes due to RFI in the context of high starch diets compared to high forage diets. An 84-day feed efficiency trial followed immediately by a second 112-day feed efficiency trial was conducted with a total of 100 animals fed either maize- or grass-diets for 196-days. At the end of the 84-day period, the 32 most divergent RFI animals (16 extreme RFI animals per diet, 8 RFI+ and 8 RFI-) were identified. They were slaughtered after 112-days of finishing. The Longissimus thoracis was characterised in terms of nutritional and sensory quality. RFI had no effect on lab colour, muscle shear force, total fat, fatty acid ratios and most of the total fatty acid content (especially n-3) irrespective of the diet. However, more efficient animals (RFI-) showed higher CLA contents compared to less efficient animals (RFI+) regardless of the diet and also a lower n6/n3 ratio only in animals fed the maize diets. Diet also had a significant effect on lipid and FA content as well as on FA composition.

Regression models from portable NIR spectra for predicting the carcass traits and meat quality of beef cattle.

The aims of this study were to predict carcass and meat traits, as well as the chemical composition of the 9th to 11th rib sections of beef cattle from portable NIR spectra. The 9th to 11th rib section was obtained from 60 Nellore bulls and cull cows. NIR spectra were acquired at: P1 -center of Longissimus muscle; and P2 -subcutaneous fat cap. The models accurately estimated (P ≥ 0.083) all carcass and meat quality traits, except those for predicting red (a*) and yellow (b*) intensity from P1, and 12th-rib fat from P2. However, precision was highly variable among the models; those for the prediction of carcass pHu, 12th rib fat, toughness from P1, and those for 12th rib fat, a* and b* from P2 presented high precision (R2 ≥ 0.65 or CCC ≥ 0.63), whereas all other models evaluated presented moderate to low precision (R2 ≤ 0.39). Models built from P1 and P2 accurately estimated (P ≥ 0.066) the chemical composition of the meat plus fat, bones and, meat plus fat plus bones, except those for predicting the ether extract (EE) and crude protein (CP) of bones and the EE of Meat plus bones fraction from P2. However, precision was highly variable among the models (-0.08 ≤ R2 ≤ 0.86) of the 9th and 11th rib section. Those models for the prediction of dry matter (DM) and EE of the bones from P1; of EE from P1; and of EE, mineral matter (MM), CP from P2 of meat plus fat plus bones presented high precision (R2 ≥ 0.76 or CCC ≥ 0.62), whereas all other models evaluated presented moderate to low precision (R2 ≤ 0.45). Thus, models built from portable NIR spectra acquired at different points of the 9th to 11th rib section were recommended for predicting carcass and muscle quality traits as well as for predicting the chemical composition of this section of beef cattle. However, it is noteworthy, that the small sample size was one of the limitations of this study.

Isolation of Nuclei from Human Intermuscular Adipose Tissue and Downstream Single-Nuclei RNA Sequencing.

Intermuscular adipose tissue (IMAT) is a relatively understudied adipose depot located between muscle fibers. IMAT content increases with age and BMI and is associated with metabolic and muscle degenerative diseases; however, an understanding of the biological properties of IMAT and its interplay with the surrounding muscle fibers is severely lacking. In recent years, single-cell and nuclei RNA sequencing have provided us with cell type-specific atlases of several human tissues. However, the cellular composition of human IMAT remains largely unexplored due to the inherent challenges of its accessibility from biopsy collection in humans. In addition to the limited amount of tissue collected, the processing of human IMAT is complicated due to its proximity to skeletal muscle tissue and fascia. The lipid-laden nature of the adipocytes makes it incompatible with single-cell isolation. Hence, single nuclei RNA sequencing is optimal for obtaining high-dimensional transcriptomics at single-cell resolution and provides the potential to uncover the biology of this depot, including the exact cellular composition of IMAT. Here, we present a detailed protocol for nuclei isolation and library preparation of frozen human IMAT for single nuclei RNA sequencing. This protocol allows for the profiling of thousands of nuclei using a droplet-based approach, thus providing the capacity to detect rare and low-abundant cell types.

Effects and mechanism of sub-freezing storage on water holding capacity and tenderness of beef.

In order to explore the effect of sub-freezing storage on water holding capacity and tenderness of beef, four treatments were compared in this study: sub-freezing (-7 °C) fast sub-freezing (-38 °C until the core temperature achieved to -7 °C), superchilling (-1 °C) and fast frozen (-38 °C until the core temperature achieved to -18 °C) with the latter two treatments serving as the controls. The differences in muscle fiber structure, water distribution, protein oxidation and cytoskeletal protein degradation were studied. The results demonstrated that compared with other treatments, the fast sub-freezing treatment resulted in less structural damage to the muscle fibers and had better water holding capacity. Both sub-freezing and fast sub-freezing treatments inhibited protein oxidation compared with superchilling, but the former treatment's level of protein oxidation was higher than that in fast sub-freezing treatment during long-term storage (42 weeks). In addition, the structural proteins in the sub-freezing and fast sub-freezing treatments underwent faster degradation during long-term storage and therefore the meat was more tender compared with the fast frozen treatment. The results indicate that the fast sub-freezing treatment can be potentially applied in beef storage.

Postmortem mitochondria function in longissimus lumborum of Angus and Brahman steers.

Mitochondria function and integrity may impact postmortem metabolism and meat quality development. Adaptations in heat tolerant Brahman may persist to limit cellular stress postmortem. Our objective was to evaluate glycolysis, pH decline, and mitochondria function in longissimus lumborum (LL) from Angus and Brahman steers (N = 28) early postmortem (1 to 6 h) and after rigor (24 h). We evaluated metabolites of anaerobic glycolysis, ATP, pH, and temperature, and determined mitochondria oxygen consumption rate (OCR) in permeabilized fibers. The main effects of breed (b) and time (t) and the interaction were tested. Brahman LL contained greater ATP during the first 6 h postmortem; Brahman also tended to exhibit a slower pH decline (b × t, P = 0.07) and more rapid temperature decline (b × t, P < 0.001), but metabolites of anaerobic glycolysis were not different. Mitochondria in Brahman and Angus LL were well-coupled and respired at 1 h postmortem. However, outer membrane integrity became increasingly compromised postmortem (t, P < 0.001). Brahman tended to exhibit greater electron transport system capacity (b, P < 0.1) and had greater capacity for oxidative phosphorylation (complex I and II substrates) at 6 h compared with Angus (P < 0.001). In totality, greater ATP, slower pH decline, and enhanced mitochondria capacity indicate that Brahman possess mitochondrial properties or cellular adaptations that help protect the cell during energy stress postmortem. Slower pH and more rapid temperature decline in LL from Brahman may also help preserve mitochondria function postmortem.

Changes in the amount of nucleotide sugars in aged mouse tissues.

Aging affects tissue glycan profiles, which may alter cellular functions and increase the risk of age-related diseases. Glycans are biosynthesized by glycosyltransferases using the corresponding nucleotide sugar, and the availability of nucleotide sugars affects glycosylation efficiency. However, the effects of aging on nucleotide sugar profiles and contents are yet to be elucidated. Therefore, this study aimed to investigate the effects of aging on nucleotide sugars using a new LC-MS/MS method. Specifically, the new method was used to determine the nucleotide sugar contents of various tissues (brain, liver, heart, skeletal muscle, kidney, lung, and colon) of male C57BL/6NCr mice (7- or 26-month-old). Characteristic age-associated nucleotide sugar changes were observed in each tissue sample. Particularly, there was a significant decrease in UDP-glucuronic acid content in the kidney of aged mice and a decrease in the contents of several nucleotide sugars, including UDP-N-acetylgalactosamine, in the brain of aged mice. Additionally, there were variations in nucleotide sugar profiles among the tissues examined regardless of the age. The kidneys had the highest concentration of UDP-glucuronic acid among the seven tissues. In contrast, the skeletal muscle had the lowest concentration of total nucleotide sugars among the tissues; however, CMP-N-acetylneuraminic acid and CDP-ribitol were relatively enriched. Conclusively, these findings may contribute to the understanding of the roles of glycans in tissue aging.

Carcass characteristics and meat quality of goat kids according to the Colomer - Rocher carcass fatness and conformation classes.

Classifying kid carcasses according to their fatness and conformation scores guides producers toward higher quality and income-generating production methods and determines the ideal slaughter time. This study aimed to determine the effects of Colomer-Rocher fatness and conformation classes on carcass and meat quality characteristics in goat kids. A total of 102 male kid carcasses were used in the study. Carcasses were divided into fatness (1-, 1, 1+, 2-) and conformation (P-, P, P+; O-) classes according to the Colomer-Rocher classification, and these groups were accepted as the experimental group. Hierarchical clustering analysis divided the kid carcasses into 5 clusters using certain carcass characteristics. Differences between clusters in most of the carcass characteristics were significant. The difference between the cluster groups in terms of meat colour and sensory characteristics was also significant. Fatness and conformation classes significantly affected most characteristics except kidney knob and channel fat (KKCF) percentages and carcass joints percentages. The Colomer-Rocher conformation classification was found to be more discriminatory in terms of meat quality than the fatness classification.

The Meat Standards Australia carcass grading site affects assessment of marbling and prediction of meat-eating quality in growing European beef cattle.

The lack of consumer feedback on beef eating quality contributes to reduced beef consumption in Europe. The Meat Standards Australia (MSA) grading scheme can assess the palatability of beef carcasses usually graded at the 10th thoracic vertebrae. However, the European beef industry relies on late-maturing breeds usually cut at the 5th vertebrae due to commercial reasons. Data from 55 young bulls and heifers of late-maturing breeds were collected in an Italian slaughterhouse following the MSA guidelines at both carcass grading sites and sides. Intramuscular fat levels were assessed through two scores and used with other variables to feed the MSA model, which predicts the MSA index, the meat-eating quality scores (MQ4) for 5 muscles and for each carcass grading site × side combination. The scores were analyzed using a mixed linear model. A correlation analysis was conducted to predict the variables measured at the 10th site using their correspondent at the 5th carcass grading site. A stepwise regression was conducted to understand the weight of each measured variable on marbling and MQ4 scores measured both at 5th and 10th carcass grading sites. Results showed significantly higher value for the studied traits at the 5th carcass grading site, while carcass side had no significant impact. The equations had high predictive capability and MSA marbling score played a key role in explaining the variability across carcass grading sites. The differences in marbling and MQ4 scores between the carcass grading sites suggest considering this factor if the MSA grading system will be applied to Europe.

Beef-on-dairy: Meat quality of veal and prediction of intramuscular fat using the Q-FOM™ Beef camera at the 5th-6th thoracic vertebra.

This study aims to describe the meat quality of young Holstein (HOL) beef-on-dairy heifers and bulls sired by Angus (ANG, n = 109), Charolais (CHA, n = 101) and Danish Blue (DBL, n = 127), and to investigate the performance of the handheld vision-based Q-FOM™ Beef camera in predicting the intramuscular fat concentration (IMF%) in M. longissimus thoracis from carcasses quartered at the 5th-6th thoracic vertebra. The results showed significant differences between crossbreeds and sexes on carcass characteristics and meat quality. DBL × HOL had the highest EUROP conformation scores, whereas ANG × HOL had darker meat with higher IMF% (3.52%) compared to CHA × HOL (2.99%) and DBL × HOL (2.51%). Bulls had higher EUROP conformation scores than heifers, and heifers had higher IMF% (3.70%) than bulls (2.31%). These findings indicate the potential for producing high-quality meat from beef-on-dairy heifers and ANG bulls. The IMF% prediction model for Q-FOM performed well with R2 = 0.91 and root mean squared error of cross validation, RMSECV = 1.33%. The performance of the prediction model on the beef-on-dairy veal subsample ranging from 0.9 to 7.4% IMF had lower accuracy (R2 = 0.48) and the prediction error (RMSEveal) was 1.00%. When grouping beef-on-dairy veal carcasses into three IMF% classes (2.5% IMF bins), 62.6% of the carcasses were accurately predicted. Furthermore, Q-FOM IMF% predictions and chemically determined IMF% were similar for each combination of sex and crossbreed, revealing a potential of Q-FOM IMF% predictions to be used in breeding, when aiming for higher meat quality.

Effect of pulsed electric field (PEF) on NaCl diffusion in beef and consequence on meat quality.

The impact of various field strength (2, 3, 4 kV/cm) and treatment time (60s and 90s) combinations on NaCl content and diffusion coefficient of beef were evaluated in the current study. Weight change, water content, water holding capacity, and texture of beef after brining were also explored. The results demonstrated pulsed electric field (PEF) pre-treatment significantly increased NaCl uptake when the brining time was 150 min (P < 0.05). The maximum NaCl content increased by 19.50% and the diffusion coefficient increased by 58.50%. Relatively mild PEF (60s) could improve beef qualities, but longer treatment time (90s) was detrimental to these qualities. Meanwhile, more complete myofibrillar structure and lower lipid oxidation extent were observed in the samples treated by PEF, contributing to the higher a* values. In conclusion, short processing time (60s) and high field strength (4 kV/cm) treatment is a potential strategy for meat brining acceleration and quality improvement in practical industrial production.

Effect of a novel steak fabrication method by trimming subcutaneous and intermuscular fats on palatability and calorie content of beef ribeye steaks.

This study investigated the effects of a novel steak fabrication method, involving removal of both subcutaneous and intermuscular fats between the longissimus thoracis (LT) and spinalis dorsi (SD) muscles before cooking, on beef ribeye steak palatability and calorie content. Canada AA (n = 10) and AAA (n = 10) ribeyes were fabricated into steaks either with (fat-on) or without (fat-off) subcutaneous and intermuscular fats. Fat-on steaks had shorter cooking times and lower cooking losses than fat-off steaks (P < 0.001), regardless of quality grade. There were treatment × quality grade interactions for initial (P < 0.01) and sustainable juiciness (P < 0.05) of the LT samples and initial juiciness (P < 0.05) of the SD samples, with the fat-on AA samples being more juicy than the fat-off AA samples, but the AAA treatments were not different from each other. Regardless of quality grade, fewer panelist responses indicated livery flavour (P < 0.05) for fat-on compared to fat-off LT samples. Regardless of fat-on/fat-off treatment, AA compared to AAA samples had more responses for bloody/serumy (P < 0.01) and unidentified off-flavours (P < 0.05) in LT samples. However, AAA samples had more responses for metallic off-flavour in LT (P = 0.059) and SD (P < 0.05). There were no differences in calorie content between fat-on compared to fat-off steaks (P > 0.1) regardless of quality grade and muscle type, or between AA and AAA steaks regardless of cooking with fat-on or off (P > 0.1). Maintaining the subcutaneous and intermuscular fats while cooking will improve ribeye steak palatability without increasing calorie content, especially for leaner steaks.

A deep learning-based approach for fully automated segmentation and quantitative analysis of muscle fibers in pig skeletal muscle.

Muscle fiber properties exert a significant influence on pork quality, with cross-sectional area (CSA) being a crucial parameter closely associated with various meat quality indicators, such as shear force. Effectively identifying and segmenting muscle fibers in a robust manner constitutes a vital initial step in determining CSA. This step is highly intricate and time-consuming, necessitating an accurate and automated analytical approach. One limitation of existing methods is their tendency to perform well on high signal-to-noise ratio images of intact, healthy muscle fibers but their lack of validation on more complex image datasets featuring significant morphological changes, such as the presence of ice crystals. In this study, we undertake the fully automatic segmentation of muscle fiber microscopic images stained with myosin adenosine triphosphate (mATPase) activity using a deep learning architecture known as SOLOv2. Our objective is to efficiently derive accurate measurements of muscle fiber size and distribution. Tests conducted on actual images demonstrate that our method adeptly handles the intricate task of muscle fiber segmentation, yielding quantitative results amenable to statistical analysis and displaying reliability comparable to manual analysis.