A single-cell atlas of bovine skeletal muscle reveals mechanisms regulating intramuscular adipogenesis and fibrogenesis.

BACKGROUND: Intramuscular fat (IMF) and intramuscular connective tissue (IMC) are often seen in human myopathies and are central to beef quality. The mechanisms regulating their accumulation remain poorly understood. Here, we explored the possibility of using beef cattle as a novel model for mechanistic studies of intramuscular adipogenesis and fibrogenesis. METHODS: Skeletal muscle single-cell RNAseq was performed on three cattle breeds, including Wagyu (high IMF), Brahman (abundant IMC but scarce IMF), and Wagyu/Brahman cross. Sophisticated bioinformatics analyses, including clustering analysis, gene set enrichment analyses, gene regulatory network construction, RNA velocity, pseudotime analysis, and cell-cell communication analysis, were performed to elucidate heterogeneities and differentiation processes of individual cell types and differences between cattle breeds. Experiments were conducted to validate the function and specificity of identified key regulatory and marker genes. Integrated analysis with multiple published human and non-human primate datasets was performed to identify common mechanisms. RESULTS: A total of 32 708 cells and 21 clusters were identified, including fibro/adipogenic progenitor (FAP) and other resident and infiltrating cell types. We identified an endomysial adipogenic FAP subpopulation enriched for COL4A1 and CFD (log2FC = 3.19 and 1.92, respectively; P < 0.0001) and a perimysial fibrogenic FAP subpopulation enriched for COL1A1 and POSTN (log2FC = 1.83 and 0.87, respectively; P < 0.0001), both of which were likely derived from an unspecified subpopulation. Further analysis revealed more progressed adipogenic programming of Wagyu FAPs and more advanced fibrogenic programming of Brahman FAPs. Mechanistically, NAB2 drives CFD expression, which in turn promotes adipogenesis. CFD expression in FAPs of young cattle before the onset of intramuscular adipogenesis was predictive of IMF contents in adulthood (R2  = 0.885, P < 0.01). Similar adipogenic and fibrogenic FAPs were identified in humans and monkeys. In aged humans with metabolic syndrome and progressed Duchenne muscular dystrophy (DMD) patients, increased CFD expression was observed (P < 0.05 and P < 0.0001, respectively), which was positively correlated with adipogenic marker expression, including ADIPOQ (R2  = 0.303, P < 0.01; and R2  = 0.348, P < 0.01, respectively). The specificity of Postn/POSTN as a fibrogenic FAP marker was validated using a lineage-tracing mouse line. POSTN expression was elevated in Brahman FAPs (P < 0.0001) and DMD patients (P < 0.01) but not in aged humans. Strong interactions between vascular cells and FAPs were also identified. CONCLUSIONS: Our study demonstrates the feasibility of beef cattle as a model for studying IMF and IMC. We illustrate the FAP programming during intramuscular adipogenesis and fibrogenesis and reveal the reliability of CFD as a predictor and biomarker of IMF accumulation in cattle and humans.

Carcass Yields and Physical-Chemical Meat Quality Characteristics of Namibian Red Hartebeest (Alcelaphus buselaphus) as Influenced by Sex and Muscle.

This study determined the carcass yields of red hartebeest from Namibia and compared the physical-chemical meat quality characteristics of six different muscles (biceps femoris, infraspinatus, longissimus thoracis et lumborum, semimembranosus, semitendinosus, and supraspinatus) for both males and females. Red hartebeest males were heavier (133.92 kg) than females (114.20 kg) but the average dressing percentage did not differ between the two sexes. Muscles from females had a lower mean shear force value of 3.59 kg/1.27 cm ø, compared to males (4.23 kg/1.27 cm ø). The most tender muscle was the infraspinatus of the female treatment group, while the semimembranosus of the male treatment group was the least tender muscle. Drip loss, cooking loss and L* (lightness) values were not affected by sex. The largest hue angle was observed in the semitendinosus muscle of the female treatment group (28.94°), and it was thus the lightest red muscle. The highest chroma values (17.3) were observed in the semimembranosus muscle. Muscle protein content averaged 20.5% over all treatment combinations, and the mean intra-muscular fat content for both male and female muscles was low (2.4%). The shoulder muscles, infraspinatus and supraspinatus, of the females had the highest fat content (2.7%). The results indicate that red hartebeest meat should be market according to specific muscles and that sex of the animals need not be considered during marketing.

Carcass yields and physiochemical meat quality characteristics of Namibian gemsbok (Oryx gazella) as influenced by muscle, gender and age.

The carcass yields and physiochemical meat quality characteristics of six different muscles (biceps femoris, infraspinatus, longissimus thoracis et lumborum, semimembranosus, semitendinosus, supraspinatus) for different sex and age groups of gemsbok antelope were determined. No live weight nor dressing percentage differences were observed for the different sex groups. While muscles, age and sex had an influence on the physiochemical parameters investigated, these differences were minor. The CIE Lab colour values for all muscles, age and sexes were in line with those deemed acceptable for game meat, while mean crude protein (20.7%) and fat (1.6%) levels were typical of game species. Discriminant analysis revealed no differentiation among the muscle groups for the variables measured. Thus, at similar live weights, male and female (sub-adult) gemsbok can be considered of equivalent meat quality and marketed accordingly.

GROWTH AND DEVELOPMENT SYMPOSIUM: STEM AND PROGENITOR CELLS IN ANIMAL GROWTH: The regulation of beef quality by resident progenitor cells1.

The intramuscular adipose tissue deposition in the skeletal muscle of beef cattle is a highly desired trait essential for high-quality beef. In contrast, the excessive accumulation of crosslinked collagen in intramuscular connective tissue contributes to beef toughness. Recent studies revealed that adipose tissue and connective tissue share an embryonic origin in mice and may be derived from a common immediate bipotent precursor in mice and humans. Having the same linkages in the development of adipose tissue and connective tissue in beef, the lineage commitment and differentiation of progenitor cells giving rise to these tissues may directly affect beef quality. It has been shown that these processes are regulated by some key transcription regulators and are subjective to epigenetic modifications such as DNA methylation, histone modifications, and microRNAs. Continued exploration of relevant regulatory pathways is very important for the identification of mechanisms influencing meat quality and the development of proper management strategies for beef quality improvement.

Advancements in meat packaging.

Packaging of meat provides the same or similar benefits for raw chilled and processed meats as other types of food packaging. Although air-permeable packaging is most prevalent for raw chilled red meat, vacuum and modified atmosphere packaging offer longer shelf life. The major advancements in meat packaging have been in the widely used plastic polymers while biobased materials and their integration into composite packaging are receiving much attention for functionality and sustainability. At this time, active and intelligent packaging are not widely used for antioxidant, antimicrobial, and other functions to stabilize and enhance meat properties although many options are being developed and investigated. The advances being made in nanotechnology will be incorporated into food packaging and presumably into meat packaging when appropriate and useful. Intelligent packaging using sensors for transmission of desired information and prompting of subsequent changes in packaging materials, environments or the products to maintain safety and quality are still in developmental stages.

Where is MAP Going? A review and future potential of modified atmosphere packaging for meat.

Modified atmosphere packaging (MAP) is the removal and/or replacement of the atmosphere surrounding the product before sealing in vapor-barrier materials. While technically different, many forms of MAP are also case-ready packaging, where meat is cut and packaged at a centralized location for transport to and display at a retail store. Most of the shelf life properties of meat are extended by use of MAP, but anoxic forms of MAP without carbon monoxide (CO) do not provide bloomed red meat color and MAP with oxygen (O(2)) may promote oxidation of lipids and pigments. Advances in plastic materials and equipment have propelled advances in MAP, but other technological and logistical considerations are needed for successful MAP systems for raw chilled fresh meat. Current MAP options of air-permeable overwrapped trays in master packs, low O(2) formats of shrunk film vacuum packaging (VP) or MAP with carbon dioxide (CO(2)) and nitrogen (N(2)) and their peelable barrier film derivatives, and high O(2) MAP each have advantages and disadvantages. Packaging technology innovations and ingenuity will continue to provide MAP that is consumer oriented, product enhancing, environmentally responsive, and cost effective, but continued research and development by the scientific and industry sectors will be needed.

Growth of Selected Cross-Contaminating Bacterial Pathogens on Beef and Fish at 15 and 35°C.

Isolates of Escherichia coli and Clostridium perfringens from beef and Aeromonas hydrophila from fish were examined for their ability to survive and grow as cross-contaminates on nonnative tissues at simulated ambient (35°C) and aging/conditioning (15°C) temperatures of handling and retailing found in the tropics. Growth of all isolates over a 10-h period was greater (P < 0.05) on their native tissues at both temperatures. The aging/conditioning temperature effectively limited growth of E. coli and A. hydrophila to less than l-logl0 CFU/g and prevented growth of C. perfringens on beef and fish samples. All three isolates demonstrated characteristic mesophilic growth response on both tissues at 35°C during the 10-h retail period. The study suggests that two muscle food products could be jointly handled to efficiently use available storage/haulage capacity in tropical countries. Potential savings in space, labor and energy would be made if cross-contamination between the two products is minimized by available packaging and sanitizing technologies.

A Simple and Effective Method for Obtaining Aseptic Raw Beef Tissues 1.

The need for gnotoxenic studies (defined microflora) in beef to better understand the microbial ecology of spoilage and pathogenic organisms necessitates the development of a simple and rapid method devoid of obvious disadvantages of existing ones. The aseptic technique described here is an improvement on the flaming and hot-iron searing methods already in use. It involved the use of a Sensa-Temp Heat Control Electric Frier (West Bend, IN) at 232°C to destroy practically all surface microflora on top round meat blocks of freshly slaughtered, healthy beef cattle, before aseptically removing the cooked exterior to a depth of approximately 1 cm. In two separate trials, 95 and 90%, respectively, of the total number of samples removed were sterile. We recommend this method to procure sterile samples suitable for inoculation with defined microflora and for other studies. The method seems applicable to a wide variety of muscle food types, including fish.