Meat Products Research Articles

Cultural context shapes the carbon footprints of recipes

Food systems are responsible for a third of global anthropogenic greenhouse gas emissions central to global warming and climate change. Increasing awareness of the environmental impact of food-centric emissions has led to the carbon footprint quantification of food products. However, food consumption is influenced by traditional dishes, the cultural capsules that encode traditional protocols for culinary preparations. Carbon footprint estimation of recipes will provide actionable insights into the environmental sustainability of culturally influenced patterns in recipe compositions. We implemented language models to integrate the carbon footprint data of food products with a gold standard repository of recipes to investigate the carbon load of recipes. Our results highlight the role of culture in shaping the carbon load of recipes. South American dishes have high carbon footprints owing to the high culinary utility of dairy and meat products. On the contrary, dishes from the Indian Subcontinent are the most environmentally friendly, with low carbon footprints due to the higher use of vegetables. While emphasizing the widely understood harms of animal-sourced ingredients, this article presents a nuanced perspective on the environmental impact of culturally influenced dietary practices. The study points to the need for efforts to develop techniques for plant-based and lab-grown meat to meet the protein requirements. We present an extensive repository of the estimated carbon footprints of recipes from across the world and enable their exploration through a user-friendly web server, SustainableFoodDB. Going forward, our study points to the need for high spatial resolution data on carbon footprints of ingredients.

A PLIN1 polymorphism is associated with fat production in male emus

The emu (Dromaius novaehollandiae) is a novel poultry species that produces meat, eggs, and fat. Although emus have recently been domesticated, genetic improvements to establish strains have scarcely progressed. In this study, we investigated the relationship between production traits and perilipin 1-encoding gene (PLIN1) polymorphisms in the emus. We determined the partial complementary DNA (cDNA) sequence of the PLIN1, which is involved in lipid droplet formation. We identified four nucleotide substitution sites (c.270C>T, c.321T>C, c.587A>T, and c.639C>T) in the PLIN1 gene of emus. Of these, c.587A>T is a non-synonymous substitution that converts lysine to methionine at the 196th codon (p.K196M). Although p.K196M was predicted to affect the production traits of emus, a large deflection in genotype frequency was observed in this study; thus, we could not investigate the relationship between genotypes and phenotypes. In males, the fat yields of the CC, CT, and TT genotypes in c.270C>T were 0.25 ± 0.06, 0.22 ± 0.06, and 0.21 ± 0.07 kg, respectively, while the meat yields of the CC, CT, and TT genotypes in c.270C>T were 0.15 ± 0.01, 0.16 ± 0.02, and 0.16 ± 0.03 kg, respectively. These results indicate that male emus with the CC genotype had a significantly higher fat content and lower meat productivity than male emus with the other genotypes (P < 0.05). Therefore, c.270C>T in PLIN1 affects fat and meat production in males. Our findings may contribute to the effective genetic improvement of the emus.

Assessing the Biogas Potential of Slaughterhouse Waste in Tenerife: A Sustainable Approach to Waste Management in an Outermost European Region

The Canary Islands, as an outermost region of Europe with fragmented territory, benefit from an exemption under Commission Regulation (EU) No 142/2011 allowing for animal by-products disposal in landfills. However, this practice has resulted in environmental and economic challenges, including reduced landfill capacity, increased greenhouse gas emissions, and missed opportunities for renewable energy production. Specifically, waste management in Tenerife faces significant challenges due to the burgeoning meat production and subsequent waste generation in its primary slaughterhouses. With an average annual production of 5,000 tons of waste, landfill capacity is nearing its limit. This study therefore aims to assess the biogas production potential from slaughterhouse waste in Tenerife and evaluate anaerobic digestion as an alternative approach. Various combinations of viscera, blood, and slaughterhouse sludge from cattle, pigs, goats, sheep, rabbits, and chickens were analyzed to determine their biogas potential. Results indicate that comprehensive valorization of these wastes could generate 12.86 GWh of energy, equivalent to approximately 20% of the energy consumed in the municipality's industrial sector. Moreover, this approach could reduce emissions by 5.88 Gg of CO2 equivalent and alleviate landfill pressures, thereby mitigating current waste management impacts. Given the region's reliance on imported fuels and limited territorial space, anaerobic digestion presents an environmentally and economically sustainable solution for managing slaughterhouse waste within a closed-loop system.

Whole genome sequencing of two strains of Staphylococcus simulans and their effects on the nitrosomyoglobin formation based on nitric oxide synthase

In order to evaluate the safety and functional characteristics of two potential meat starter cultures, Staphylococcus simulans HZ01 (OM758216) and Staphylococcus simulans HZ02 (OM758217), we performed the whole genome sequences, and then investigated their effects on nitrosomyoglobin formation based on nitric oxide synthase (NOS) catalysis. The results revealed that the two strains did not possess any virulence genes. The gene annotation indicated the gene function is mainly due to translation, ribosomal structure and biogenesis, and amino acid transport and metabolism. Both strains possess nos gene. The phylogenetic tree based on nos gene indicated the two strains formed an independent evolutionary branch. Subsequently, NOS was found to be expressed in both HZ01 and HZ02. NOS activity was higher in HZ01 than in HZ02, whereas NO production in inoculated groups changed considerably with increasing culture time. In the model system, nitrosomyoglobin formed in S. simulans inoculated groups, and the derivatives had absorption peaks at 545 nm and 580 nm. The secondary structure of these derivatives exhibited differences in β-turn proportion and α-helix proportion. Physicochemical properties of fermented sausages inoculated groups indicated that S. simulans HZ01 and HZ02 can significantly improve the color of sausages with no addition of nitrite/nitrate. This result confirmed that the two S. simulans strains can improve the nitrosomyoglobin formation based on the NOS pathway, which contributes to the color of fermented sausages, without the requirement of nitrite or nitrate. Nevertheless, the contribution of the NOS pathway in fermented meat products should be further evaluated.

A diarylheptanoid derivative mediates glycogen synthase kinase 3β to promote the porcine muscle satellite cell proliferation: Implications for cultured meat production

Skeletal muscle stem cells, or satellite cells, are vital for cultured meat production, driving proliferation and differentiation to form muscle fibers in vitro. However, these abilities are often compromised after long-term in vitro culturing due to a loss of their stemness characteristics. Therefore, effective pharmacological agents that enhance satellite cell proliferation and maintain stemness ability are needed for optimal cell growth for cultured meat production. In this study, the effects of the identified glycogen synthase kinase 3β (GSK3β) inhibitors, ASPP 049, a diarylheptanoid isolated from Curcuma comosa rhizomes, and CHIR 99021 on porcine muscle satellite cell (PMSC) proliferation and Wnt/β-catenin signaling pathway were investigated. We found that both compounds enhanced cell viability and proliferation while preserving the stemness marker, as evidenced by increased expression of the skeletal muscle stem cell marker, Pax7 protein. Molecular dynamics simulations showed that ASPP 049 and CHIR 99021 exhibited differing binding affinities, primarily through hydrophobic interactions, suggesting potential for the design of more potent inhibitors in the future. Despite its weaker binding, ASPP 049 still showed significant effects on the regulation of the Wnt/β-catenin signaling pathway via increased phosphorylation of GSK3β at Ser9 and decreased the phosphorylation of β-catenin at Ser33, Ser37, and Thr41, thereby subsequently activating Wnt transcriptional activity. This study highlights the potential of ASPP 049 and CHIR 99021 to enhance PMSC proliferation and maintain stemness ability, offering a promising avenue for improving cultured meat production.

Effect of ultrasonic pretreatment with synergistic microbial fermentation on tenderness and flavor of air-dried duck under low nitrite process

The tenderness and flavor of meat products are critical factors influencing consumers' purchasing decisions. This study investigated the effects of ultrasonic pretreatment with synergistic microbial strain fermentation on tenderness and flavor of air-dried duck under low nitrite process. The results demonstrated that ultrasonic pretreatment combined with microbial strain fermentation improved water retention and tenderness of duck meat by disrupting the muscle microstructure, increasing muscle fiber spacing, and facilitating water migration and distribution. This primarily concerns the cavitation and mechanical effects of ultrasound and the role of lactic acid bacteria and yeasts in muscle protein hydrolysis. A total of 34 and 55 volatile flavor compounds were detected by HS-SPME-GC–MS and GC-IMS, respectively. The results indicated that acetaldehyde (stimulating, fruity, green apple), ethyl acetate (sweet, fruity, pineapple), and 3-hydroxy-2-butanone (sweet, creamy) were responsible for the improved flavor during this process, which was primarily related to the increased activity of neutral lipase (0.38 U/g protein), acidic lipase (0.48 U/g protein), and phospholipase (0.09 U/g protein). This study provides valuable insights into the synergistic effects of ultrasonic pretreatment and microbial co-fermentation, offering a theoretical basis for optimizing air-dried duck production and enhancing flavor quality.

Effect of pre-acidification induction on the physicochemical features, myofibrillar protein microstructure, and headspace volatiles of ready-to-cook goose meat

This study examined the impact of pre-acidification induction on the quality attributes and flavor retention of ready-to-cook (RTC) goose meat products. The results demonstrated that pre-acidification could influence the eating qualities of RTC goose meat by effectively regulating the physicochemical properties of goose myofibrillar proteins (MP) including solubility and water-holding capacity. Elevated carbonyl content indicated an enhanced gel-forming capacity in RTC goose meat during storage, coupled with reduced total sulfhydryl content from enhanced protonation pretreatment and augmented lipid oxidation. Structural characterization of MP via sodium dodecyl sulfate–polyacrylamide gel electrophoresis, circular dichroism spectroscopy, and intrinsic fluorescence revealed the formation of a dense protein matrix under highly acidic conditions. Furthermore, the headspace concentration of aldehydes increased by 3.23 times upon enhancing the pre-acidification intensity, resulting in the production of esters and acidic flavor compounds with favorable aromas. Correlation analysis demonstrated the dependence of headspace concentrations of volatile constituents on the acidification-enhanced surface hydrophobicity of MP, attributed to the modified binding sites of proteins after pre-acidification. Current results have indicated both the positive and negative influence of pre-acidulation induction on the eating quality of goose meat products, suggesting the necessity of introducing extra processes to modulate the quality of prefabricated products.

Abelmoschus manihot gum improves the water retention capacity of low-salt myofibrillar protein gels: Perspective on aggregation behaviour and conformational changes during heating

This study aimed to investigate the effect of Abelmoschus manihot gum (AMG) on the water retention capacity of low-salt myofibrillar protein (MP) gel by analysing its aggregation behaviour and conformational changes during heating (30–80 °C). The results revealed that AMG significantly increased the water holding capacity and facilitated the formation of a more uniform gel network structure in low-salt MP gel (P < 0.05). During the heat-induced gelation process, the solubility of low-salt MP significantly decreased, whereas its turbidity evidently increased as the level of added AMG increased (P < 0.05). Furthermore, the dynamic rheological behaviours indicated that low-salt MP-AMG gels underwent early denaturation and unfolded at 58 °C, finally forming an irreversible three-dimensional network at 80 °C. Moreover, adding AMG promoted α-helix-to-β-sheet transition in low-salt MP and decreased its fluorescence intensity during the heating process. Hydrophobic interactions and disulfide bonds were the two dominant forces governing the formation and maintenance of low-salt MP gel. The present study provides theoretical guidance for the production of novel low-salt healthy meat products.

Enhacing emulsification of meat broth system mixed with myofibrillar proteins and type I collagen: The role of NaCl and heat

A mixed system with a 5:1 ratio of beef myofibrillar protein to type I collagen was prepared to mimic meat broths. The study aimed to determine the combined effects of various NaCl concentrations (0, 0.2 M, 0.4 M, 0.6 M) and heat treatment on solubility, emulsifying properties (EAI, ESI), viscosity, and particle size of the mixed protein system. Mechanistic changes were examined through molecular interactions, intrinsic fluorescence, protein molecular weight, and Raman spectroscopy. The results showed that, without heat treatment, NaCl enhanced solubility, EAI, ESI, emulsion viscosity, and hydrogen bonding. After heating (90 °C, 30 min), elevated 0.4–0.6 M NaCl created an unstable, crowded environment, resulting in protein aggregation and reduced solubility and emulsifying performance. The results indicated that heating at 90 °C with 0.2 M NaCl was beneficial for meat emulsification, providing valuable production guidance for optimizing the formulation of meat products with low salt and high emulsifying properties.

Effects of guar gum/chitosan edible films functionalized with citronellal/HPβCD inclusion complex on Harbin red sausage preservation

Harbin red sausage is a traditional local pork meat product in China, but it is susceptible to microbial contamination and lipid oxidation, leading to quality deterioration. Herein, guar gum (GG)/chitosan (CS) edible films functionalized with citronellal/hydroxypropyl-β-cyclodextrin inclusion complex (CIT/HPβCD-IC) were fabricated for Harbin red sausage preservation. Results demonstrated CIT/HPβCD-IC was successfully prepared and observed by SEM due to the bathochromic shift of maximum absorption peak of CIT, and the formation of new bonds was confirmed by FTIR analysis, suggesting the embedding of CIT into HPβCD cavity. The changes of functional groups stretching vibrations suggested successful loading of CIT/HPβCD-IC into the GG/CS edible films. Furthermore, the incorporation of CIT/HPβCD-IC enhanced the microstructural, mechanical and barrier properties, and improved the antibacterial activities, biodegradability and thermal stability of the GG/CS edible films. Particularly, the GG/CS edible films incorporated with 1 % CIT/HPβCD-IC (GG/CS-IC 1 %) enhanced the storage stability of Harbin red sausage at 4 °C by decreasing the weight loss rate, maintaining the pH, color, and textural stabilities, retarding the microbial growth and lipid oxidation of the sausage samples. Findings here suggested that GG/CS-IC 1 % edible films showed great potential as novel multi-functional edible packaging materials for Harbin red sausage preservation.

Assessment of the risk of Staphylococcus aureus infection associated with the consumption of braised beef meat sold on the streets in Côte d'Ivoire

Braised beef meat sold on the streets, a meat product that poses a potential risk of food poisoning to humans. This study was to assess the risk of foodborne infection by S. aureus associated with the consumption of braised beef in Côte d’Ivoire, to improve food safety in this country. A preliminary survey was first carried out in six (06) separate towns in Cȏte d'Ivoire, among consumers (n = 900) and sellers (n = 300) of braised beef meat with a view to characterizing the behavior of these actors. A microbiological analysis was then carried out in accordance with the ISO 6888-1/-3 standard (1999) on 189 samples of this braised beef meat taken from various sales outlets in these towns. A risk model was developed. The risk of infection linked to the consumption of this braised beef was estimated using the Monte Carlo simulation procedure. The results of the consumer survey showed that the percentage of the population consuming braised beef meat (BBM) was 74.4%, with an average consumption of 114.3 ± 0.5 g/person/day. Microbiological analysis revealed the isolation of 92% S. aureus with a mean load of 6.0 ± 0.19 log10∙cfu/g greater than 105 CFU/g. The probability of ingesting a dose greater than 108 S. aureus bacteria ranged from 4.2% to 4.3%. Braised beef meat sold in the streets of Côte d'Ivoire's towns and cities poses a real risk of infection. S. aureus is one of the causes. S. aureus-associated gastroenteritis is caused by failure to observe simple hygiene rules. The risk of infection should be mitigated by cleaning up the places where this product is sold and promoting good hygiene practices in the informal sector.

A Nitbit Citation of the Contributions of Sheikh Abdullahi Fodiyo in the Public Health of The 19th Century West Africa and Beyond: A Case Study of "Kitaabu Niyyati Fil' A'amaliddunyawiyyati Wal'ddiniyyati" (A Book of Intentions in the Issues of World and

Nowadays, invariably health improvement, health education, health benefits are attached to religiosity as confirmed by inexhaustible literatures. This paper reviewed some of the contributions of Sheikh Abdullahi Fodiyo in the public health of 19th century West African Sokoto Caliphate and afterwards in his famous book called "Kitaabu Niyyati Fil' A'amaliddunyawiyyati Wal'ddiniyyati" (A book of intentions in the issues of world and religion). In the examined book, he stressed the importance of cleanliness, ablution, prayer, work/ employment/ wealth, agriculture, and meat production in the development and sustainability of society, let alone the Muslim ummah. He warned against unemployment, lack of personal hygiene, poor production and handling of food/ meat among others. Certainly, the book exhorts benefits to public health especially in Islamic societies. There is need to use religion as health education, and health promotion tool especially in religious societies for better health and interventions.

Associations Between Microsatellites Markers and Growth Traits in Goat

The rapid progress in the field of biotechnology gives us a great chance to improve the productivity of our native animals in short time, relatively. The situation in Saudi Arabia not differ in compare with other country in our region which have a big challenge to increase the meat production for food security purposes. The situation in goat is very far from the point which other farm animals like cattle reached today, where a complete QTLs maps for most important traits is available. But by the same way, long time ago we have a complete comparative map between goat and both sheep and cattle. According to that in the previous investigation in both sheep and cattle we collected a total of 19 loci distributed in goat different chromosomes along with a total of 1050 records from 525 individual of a crossbred of Aradi and Damascus Goat were used to perform the linkage analysis. The studied traits were body weight at birth, 4, 8, 12, 16, 20, and 24 weeks of age. The association between microsatellite markers and body weights traits were estimated by stepwise partial regression. Twelve markers (BMS2325, BMS332, BM17132, BMS1316, BM1827, BM1225, BMS2142, BM1558, BMS2809, BMS1348, BMC4216 and BM1329) successfully linked to studied traits. These markers are. In general, our work design and plan successfully worked and could explained some of genetic vitiation by using microsatellites markers. Further work to build a complete QTLs map for meat production traits is necessary to make a complete view about this important species.

Recent advancements in meat traceability, authenticity verification, and voluntary certification systems

The growing demand for transparency in the food industry has led to significant advancements in meat traceability. Ensuring the authenticity and origin of meat products is critical for consumer trust, public health, and compliance with regulations. This paper reviews recent innovations in meat traceability, with a focus on blockchain technology as a novel approach to ensuring traceability. Additionally, advanced methods for verifying meat authenticity and origin, such as isotope fingerprinting, DNA analysis, and spectroscopic methods, are discussed. The role of voluntary certification schemes in enhancing traceability and authenticity verification in the meat industry is also explored. The findings highlight the importance of integrating cutting-edge technologies and certification schemes to build a robust and transparent meat supply chain.

Novel Insights into the Enrichment Pattern of Aroma and Taste in Cooked Marinated Meat Products of Black Pork via Typical Process Steps

This study aims to reveal the evolution mechanism of odour and taste active compounds in cooked marinated pork knuckles via typical process steps; among them, the brine soup stage was the most important part due to spices’ enriching flavours. These results revealed that the content and diversity of volatile compounds increased due to the addition of spices and heating temperature, imparting a unique aroma. Aldehydes played the main role in the overall odour. Benzaldehyde, hexanal, 1-octen-3-ol, levulinic acid, hydroxyacetone, ethyl octanoate, and 2-pentyl-furan were identified as the most important odour-active compounds. The key taste-active amino acids were glutamine, leucine, valine, and lysine. The IMP, AMP, and GMP provided a strong umami taste. Taste nucleotides and Val, Leu, Ile, and Phe were important precursor substances for aldehydes. The high responses of the electronic nose indicated that the gas component contained alkanes, alcohols, and aldehydes. The synergistic effects between umami-free amino acids and nucleotides correlated well with umami, as assessed by the electronic tongue. These results could be a starting point for the manufacturing industry, contributing to a better understanding of product performance.

Exploring the Nexus of Feeding and Processing: Implications for Meat Quality and Sensory Perception

The intrinsic quality of meat is directly related to muscle and fat tissues. Factors such as the rate and extent of anaerobic glycolysis affect muscle pH, influencing the meat’s color, water holding, and texture. Postmortem anomalies can result in deviations from this intrinsic quality. The animals’ diet plays a crucial role in meat quality. Specific nutrients, such as proteins, vitamins, and minerals, affect meat’s texture, flavor, and juiciness. Feeds rich in omega-3 fatty acids can improve the sensorial quality of meat. Meat processing and methods such as aging, marinating, and cooking affect the texture, flavor, and juiciness, which can be evaluated by specific equipment or trained or untrained consumers. This comprehensive review investigates the relationship between animal feeding practices and meat processing techniques and their combined impact on meat quality and sensory perception. By synthesizing recent research, we explore how various feeding protocols (including diet composition and feed additives) and processing methods shape meat products’ nutritional value, texture, flavor profile, and overall consumer appeal. Understanding this nexus is crucial for optimizing meat quality while ensuring sustainability and safety in the food supply chain.

Initial Evaluation of Safety and Immunomodulatory Potential of Dietary Supplementation with Mangosteen Pericarp Extract for Sustainable Meat Production in Native Crossbred Chickens

The utilization of mangosteen biomass not only solves environmental problems but also raises the value of agricultural waste. The current study aimed to evaluate the potential of mangosteen pericarp extract (MPE) for enhancing the immunity and productivity of Thai native crossbred chickens on-farm. A total of 180 three-week-old chickens were divided into negative control and supplemented groups, with 1000 mg MPE/kg of diet. The safety of MPE was further confirmed by the absence of noticeable differences in mortality and biochemical parameters during the entire study period. The MPE-supplemented group displayed significant differences in the relative transcription levels of IL-10 compared to the basal diet group (p ≤ 0.01). Preslaughter body weight, average daily gain, and carcass weight in the MPE-supplemented group were higher than those in the basal diet group (p ≤ 0.05). Furthermore, MPE supplementation improved meat quality by enhancing the nutritional composition of protein and fat (p ≤ 0.05), as well as improving water-holding capacity, lowering boiling, and lowering grilling losses (p ≤ 0.01). These findings indicate that MPE can be an effective supplement for enhancing flock immunity, growth performance, and meat quality in poultry. This contributes to more sustainable agriculture and food security within agroecosystems.

Genetic Diversity and Population Structure Analysis in Guar

Guar [Cyamopsis tetragonoloba (L.) Taub] was domesticated in India and Pakistan. It is mainly self-pollinated, bushy, and deeply tap-rooted. Guar seed endosperm contains galactomannan gum, which is used in many food products, pharmaceuticals, cosmetics, explosives, meat products, and pet foods, and in the textile industry, yet its genetic diversity remains largely underexplored. Using 7000 high-quality single nucleotide polymorphism (SNP) markers acquired from genotyping by sequencing (GBS), we analyzed the genetic diversity and population structure in 225 guar accessions from India, Pakistan, and the United States. Structure Harvester revealed that K = 3 had the best delta K, whereas K = 2 had the second-highest delta K. Three major genetic clusters (K = 3) were identified using population structure analysis, utilizing an admixture model: 156 accessions (69.3%) were classified into Q1, 23 accessions (10.2%) in Q2, and 16 accessions (7.1%) in Q3. The remaining 30 accessions (13.3%) were included in the admixture. In all three of the subpopulations at K = 3, most of the guar accessions came from India. We also found that these clusters mostly correlated with geographic origins. Results showed that the Q2 and Q3 subpopulations included only guar accessions from India. Genetic resources from Q2 and Q3 may represent an untapped reservoir for introducing beneficial variety into the U.S. breeding populations. This genetic diversity and population structure analysis of the guar gene pool will be of interest to conduct allele mining and donor parent selection for the development of new and better guar germplasm for desired traits.

Beef Carcasses Aged at Mild Temperature to Improve Sustainability of Meat Production

Beef carcass aging, which enhances tenderness and flavor through proteolysis, is traditionally costly and slow, requiring long-term storage at temperatures near 0 °C. To reduce energy consumption, a new technique using moderate cooling room temperatures was tested. Six carcasses of Holstein bulls were used. From each carcass, two shoulders were processed in different ways: one was refrigerated at 8 °C (W), and after spraying with a solution with calcium chloride and sodium chloride, was coated with sodium alginate. The other shoulder was stored at 2 ± 1 °C as a cold control (C). After five days of aging, the shoulders were dissected, and two muscles (Caput longum triceps brachii and Supraspinatus) were subjected to physico-chemical analysis, microbiological safety assessment, and sensory testing. The remaining samples of both muscles were stored in domestic conditions for an additional 5 days at various temperatures (2, 4, 8 °C), where the same physic-chemical and sensory tests were conducted. The results showed that moderate aging temperature improved meat quality, significantly reducing the shear force (p = 0.001) and increasing sarcomere length, the myofibrillar fragmentation index, and sensory tenderness (p = 0.042, p = 0.039, and p = 0.027, respectively). However, domestic storage post-dissection should not exceed 4 °C to prevent rapid lipid oxidation, as observed at 8 °C for both muscles (p &lt; 0.001). Mild aging temperature maintained legal safety standards, enhanced certain meat qualities, and promoted enzymatic activity similar to traditional dry aging while reducing high energy consumption.

Optimisation of cell fate determination for cultivated muscle differentiation.

Production of cultivated meat requires defined medium formulations for the robust differentiation of myogenic cells into mature skeletal muscle fibres in vitro. Although these formulations can drive myogenic differentiation levels comparable to serum-starvation-based protocols, the resulting cultures are often heterogeneous, with a significant proportion of cells not participating in myofusion, limiting maturation of the muscle. To address this problem, we employed RNA sequencing to analyse heterogeneity in differentiating bovine satellite cells at single-nucleus resolution, identifying distinct cellular subpopulations including proliferative cells that fail to exit the cell cycle and quiescent 'reserve cells' that do not commit to myogenic differentiation. Our findings indicate that the MEK/ERK, NOTCH, and RXR pathways are active during the initial stages of myogenic cell fate determination, and by targeting these pathways, we can promote cell cycle exit while reducing reserve cell formation. This optimised medium formulation consistently yields fusion indices close to 100% in 2D culture. Furthermore, we demonstrate that these conditions enhance myotube formation and actomyosin accumulation in 3D bovine skeletal muscle constructs, providing proof of principle for the generation of highly differentiated cultivated muscle with excellent mimicry to traditional muscle.