Objective: The aim of this study was to select the candidate genes affecting meat quality and preliminarily explore the related molecular mechanisms in the Mashen pig. Methods: The present study explored genetic factors affecting meat quality in the Mashen pig using RNA sequencing (RNA-Seq). We sequenced the transcriptomes of 180-day-old Mashen and Large White pigs using longissimus dorsi to select differentially expressed genes (DEGs). Results: The results indicated that a total of 425 genes were differentially expressed between Mashen and Large White pigs. A gene ontology enrichment analysis revealed that DEGs were mainly enriched for biological processes associated with metabolism and muscle development, while a Kyoto encyclopedia of genes and genomes analysis showed that DEGs mainly participated in signaling pathways associated with amino acid metabolism, fatty acid metabolism, and skeletal muscle differentiation. A MCODE analysis of the protein-protein interaction network indicated that the four identified subsets of genes were mainly associated with translational initiation, skeletal muscle differentiation, amino acid metabolism, and oxidative phosphorylation pathways. Conclusion: Based on the analysis results, we selected glutamic-oxaloacetic transaminase 1, malate dehydrogenase 1, pyruvate dehydrogenase 1, pyruvate dehydrogenase kinase 4, and activator protein-1 as candidate genes affecting meat quality in pigs. A discussion of the related molecular mechanisms is provided to offer a theoretical basis for future studies on the improvement of meat quality in pigs.
Meat quality comprises a set of key traits such as pH, meat color, water-holding capacity, tenderness and marbling. These traits are complex because they are affected by multiple genetic and environmental factors. The aim of this study was to investigate the molecular genetic basis underlying nine meat quality-related traits in a Yorkshire pig population using a genome-wide association study (GWAS) and subsequent biological pathway analysis. In total, 45,926 single nucleotide polymorphism (SNP) markers from 543 pigs were selected for the GWAS after quality control. Data were analyzed using a genome-wide efficient mixed model association (GEMMA) method. This linear mixed model-based approach identified two quantitative trait loci (QTLs) for meat color (b*) on chromosome 2 (SSC2) and one QTL for shear force on chromosome 8 (SSC8). These QTLs acted additively on the two phenotypes and explained 3.92%-4.57% of the phenotypic variance of the traits of interest. The genes encoding HAUS8 on SSC2 and an lncRNA on SSC8 were identified as positional candidate genes for these QTLs. The results of the biological pathway analysis revealed that positional candidate genes for meat color (b*) were enriched in pathways related to muscle development, muscle growth, intramuscular adipocyte differentiation, and lipid accumulation in muscle, whereas positional candidate genes for shear force were overrepresented in pathways related to cell growth, cell differentiation, and fatty acids synthesis. Further verification of these identified SNPs and genes in other independent populations could provide valuable information for understanding the variations in pork quality-related traits.
This study was conducted to compare proteins expressed in M. longissimus from Hanwoo and Holstein steers immediately after slaughter. Two-dimensional electrophoresis (2DE)/LC-MS/MS analysis revealed that the total number of detectable protein spots from longissimus muscle tissues was slightly higher in Hanwoo ($575{\pm}65$) than Holstein ($534{\pm}13$) steers, but that these numbers were not statistically significant due to large variation between replicates. A total of twelve protein spots did not match between sample groups, eight of which were expressed in the Hanwoo sample and four that were expressed in the Holstein sample. The protein spots detected in the Hanwoo sample included smooth muscle and non-muscle myosin alkali light chain 6B isomers, ${\alpha}B$ crystallin isomers, hemoglobin ${\beta}$-A chains, slow myosin heavy chains, and slow skeletal muscle troponin T chains. Collectively, these proteins are a class of slow-twitch muscle fiber and mirror that Hanwoo muscle tissue sampled for the current study contained more slow-twitch muscle fibers than Holstein one. Conversely, proteins detected from the Holstein sample included ankyrin repeat domain 2 and creatin kinase isomers. Given that creatin kinase isomers are related to the fast-twitch muscle, these results likely indicate that Holstein muscle tissue sampled for the current study contained more fast-twitch muscle fibers than Hanwoo beef.
Objective: The effects of aging and freezing/thawing sequence on color, physicochemical, and enzymatic characteristics of two beef muscles (Mm. gluteus medius, GM and biceps femoris, BF) were evaluated. Methods: Beef muscles at 3 d postmortem were assigned to four different combinations of aging and freezing/thawing sequence as follows; aging at $2^{\circ}C$ for 3 wk (A3, never-frozen control), freezing at $-28^{\circ}C$ for 2 wk then thawing (F2, frozen/thawed-only), aging at $2^{\circ}C$ for 3 wk, freezing at $-28^{\circ}C$ for 2 wk then thawing (A3F2), and freezing at $-28^{\circ}C$ for 2 wk, thawing then further aging at $2^{\circ}C$ for 3 wk (F2A3). Results: No significant interactions between different aging/freezing/thawing treatments and muscle type on all measurements were found. Postmortem aging, regardless of aging/freezing/thawing sequence, had no impact on color stability of frozen/thawed beef muscles (p<0.05). F2A3 resulted in higher purge loss than F2 and A3F2 treatments (p<0.05). A3F2 and F2A3 treatments resulted in lower shear force of beef muscles compared to F2 (p<0.05). Although there was no significant difference in glutathione peroxidase (GSH-Px) activity, F2A3 had the highest ${\beta}-N-acetyl$ glucominidase (BNAG) activity in purge, but the lowest BNAG activity in muscle (p<0.05). GM muscle exhibited higher total color changes and purge loss, and lower GSH-Px activity than BF muscle. Conclusion: The results from this present study indicate that different combinations of aging/freezing/thawing sequence would result in considerable impacts on meat quality attributes, particularly thaw/purge loss and tenderness. Developing a novel freezing strategy combined with postmortem aging will be beneficial for the food/meat industry to maximize its positive impacts on tenderness, while minimizing thaw/purge loss of frozen/thawed meat.
This study examined the effects of different wet-aging times on the physicochemical characteristics and microbial profile of longissimus lumborum (LL) and biceps femoris (BF) muscles from Korean native black goat (KNBG) meat. The water holding capacity (WHC), pH, cooking loss, shear force, meat color, free amino acid, total bacteria, and coliform count of KNGB meat were analyzed at 0, 5, 10, and 15 days of wet-aging at 4℃ under vacuum packaging. The results showed that different wet-aging times led to significant pH variations between the muscles throughout the aging period. The wet-aging time did not affect the WHC and cooking loss in meat from the LL muscle. In the BF muscle, however, meat wet-aged for five days had a significantly higher WHC and less cooking loss than meat aged for 0, 10, and 15 days. The meat from the LL muscle wet-aged for five days produced tenderer meat (low shear force value) than the unaged meat (p < 0.05). Moreover, the color was similar in the LL muscle regardless of the number of aging days. In the BF muscle, the redness (a*) was higher in the meat wet-aged for 15 days compared to that aged for 0, 5, and 10 days (p < 0.05). Regardless of the muscles, an increase in wet-aging time led to an increase in the total free amino acids contents in both muscles (p < 0.05). On the other hand, the tasty/bitter amino acid ratio was significantly higher for five days of wet-aged meat than 10 and 15 days of aging from the BF muscle. In addition, regardless of the muscles, the total bacteria and coliform counts were significantly lower for five days of wet-aged meat than 10 and 15 days of aging (p < 0.05). Therefore, chevon wet-aged for five days is an optimal aging period under vacuum packaging that fortifies meat quality with a minimal microbial negative defect.
Objectives: Sarcopenia is an age-associated skeletal muscle disorder that can profoundly impact the health of elderly people. However, the efficacy of herbal medicine in sarcopenia is uncertain. This review aims to investigate evidence of the effect of herbal medicine on sarcopenia. Methods: We systematically searched 12 electronic databases for relevant randomized controlled trials (RCTs). Only trials that met the inclusion criteria were selected, and the characteristics of the included studies were extracted and synthesized in a narrative manner. The quality of the included studies was assessed using Cochrane's Risk of Bias (RoB) 2.0 tool. Results: 7 RCTs involving 672 participants with sarcopenia met the inclusion criteria. The intervention combining herbal medicine and conventional treatment (i.e., exercise, nutritional support) had a significant therapeutic effect compared with the conventional treatment, showing improvement in muscle strength (i.e., grip strength), muscle mass (i.e., appendicular skeletal muscle mass index), and physical function (i.e., gait speed, short physical performance battery, and timed up and go test). However, the methodological quality of the included RCTs was relatively low due to their high RoB, making it difficult to evaluate the efficacy of herbal medicine in sarcopenia. In terms of safety, several adverse events were reported. Conclusion: This review suggests that herbal medicine has a positive effect on muscle strength, muscle mass, and physical performance in elderly patients with sarcopenia, but there is a clear need for further research in this area.
Objectives The purpose of this study is to systematically review the effects of Neuromuscular Electrical Stimulation treatment on muscle wasting in cancer patients. Methods Randomized controlled trials (RCTs) were searched from seven online databases (PubMed, Cochrane Library, EMBASE, China National Knowledge Infrastructure, Oriental Medicine Advanced Searching Integrated System, Korean studies Information Service System, Research Information Sharing Service. The selected RCTs were evaluated for methodological quality through the Cochrane RoB. Results A total of 126 articles were identified, and 4 randomized controlled trials were selected for systematic review. In one study, it was found that there were statistically significant improvements in the Health-Related Quality of Life (FACTH&N total score, p<0.001). Additionally, significant effects were observed in measurements that represented the size of the quadriceps muscle (cross-sectional area of Vastus lateralis and Rectus femoris, p=0.004), maximum muscle strength, the twitch response of resistance muscles, and voluntary activation (p<0.001). However, no significant differences were observed between the intervention and control groups in terms of quadriceps muscle strength in two other studies (p>0.05). Lastly, while one study showed no significant differences in muscle fiber characteristics between the two groups, it did report significant improvements in measurements related to mitochondria within muscle tissue and muscle strength in the intervention group (p<0.05). Conclusions Neuromuscular electrical stimulation can be a method used to improve muscle strength in muscle wasting of cancer patients, but it is difficult to see its effects as significant compared to other treatments.
Gene expression profiling has offered new insights into postmortem molecular changes associated with meat quality. To acquire reliable transcript quantification, high quality RNA is required. The objective of this study was to analyze integrity of RNA isolated from chicken skeletal muscle (pectoralis major) and its capability of serving as the template in quantitative real-time polymerase chain reaction (qPCR) as a function of postmortem intervals representing the end-points of evisceration, carcass chilling and aging stages in chicken abattoirs. Chicken breast muscle was dissected from the carcasses (n = 6) immediately after evisceration, and one-third of each sample was instantly snap-frozen and labeled as 20 min postmortem. The remaining muscle was stored on ice until the next rounds of sample collection (1.5 h and 6 h postmortem). The delayed postmortem duration did not significantly affect $A_{260}/A_{280}$ and $A_{260}/A_{230}$ ($p{\geq}0.05$), suggesting no altered purity of total RNA. Apart from a slight decrease in the 28s:18s ribosomal RNA ratio in 1.5 h samples (p<0.05), the value was not statistically different between 20 min and 6 h samples ($p{\geq}0.05$), indicating intact total RNA up to 6 h. Abundance of reference genes encoding beta-actin (ACTB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), hypoxanthine-guanine phosphoribosyltransferase (HPRT), peptidylprolylisomerase A (PPIA) and TATA box-binding protein (TBP) as well as meat-quality associated genes (insulin-like growth factor 1 (IGF1), pyruvate dehydrogenase kinase isozyme 4 (PDK4), and peroxisome proliferator-activated receptor delta (PPARD) were investigated using qPCR. Transcript abundances of ACTB, GAPDH, HPRT, and PPIA were significantly different among all postmortem time points (p<0.05). Transcript levels of PDK4 and PPARD were significantly reduced in the 6 h samples (p<0.05). The findings suggest an adverse effect of a prolonged postmortem duration on reliability of transcript quantification in chicken skeletal muscle. For the best RNA quality, chicken skeletal muscle should be immediately collected after evisceration or within 20 min postmortem, and rapidly preserved by deep freezing.
The objective of this study was to assess the effects of constant high ambient temperatures on meat quality, antioxidant capacity, and carnosine expression in longissimus dorsi muscle of finishing pigs. Castrated 24 male DLY (crossbreeds between Landrace${\times}$Yorkshire sows and Duroc boars) pigs were allocated to one of three treatments: constant ambient temperature at $22^{\circ}C$ and ad libitum feeding (CON, n = 8); constant high ambient temperature at $30^{\circ}C$ and ad libitum feeding (H30, n = 8); and constant ambient temperature at $22^{\circ}C$ and pair-fed with H30 (PF, n = 8). Meat quality, malondialdehyde (MDA) content, antioxidant capacity, carnosine content, and carnosine synthetase (CARNS1) mRNA expression in longissimus dorsi muscle were measured after three weeks. The results revealed that H30 had lower $pH_{24h}$, redness at 45 min, and yellowness at 24 h post-mortem (p<0.05), and higher drip loss at 48 h and lightness at 24 h post-mortem (p<0.01). Constant heat stress disrupted the pro-oxidant/antioxidant balance in longissimus dorsi muscle with higher MDA content (p<0.01) and lower antioxidant capacity (p<0.01). Carnosine content and CARNS1 mRNA expression in longissimus dorsi muscle of H30 pigs were significantly decreased (p<0.01) after three weeks at $30^{\circ}C$. In conclusion, constant high ambient temperatures affect meat quality and antioxidant capacity negatively, and the reduction of muscle carnosine content is one of the probable reasons.
This experiment was conducted to compare the effects of feeding DL-2-hydroxy-4-(methylthio)butanoic acid (HMTBA) and DL-methionine (DLM) supplemented corn-soybean-cottonseed meal diets on growth performance, carcass composition, and muscle color of broilers. The trial was designed as a $2{\times}3{\times}2$ factorial experiment, including two methionine (Met) sources (HMTBA and DLM), three equimolar graded levels of Met supplementation (i.e., 0.08, 0.16, and 0.24% in the starter diet and 0.07, 0.14, and 0.21% in the grower and finisher diets, respectively), and two sexes (male and female). Additionally, one basal diet for each sex was formulated to be limiting in Met to test the dosage response of increasing supplemental Met levels. Four hundred and twenty 10-d-old broilers were randomly allotted to 14 treatments (seven each for males and females), with five replicate pens per treatment and six chicks per pen. There was no difference (p>0.05) between the two Met sources in growth performance and muscle deposition of broilers throughout the whole experimental period (d 10 to 49). With the increasing Met supplementation levels, average daily gain was increased (quadratic; p<0.01) during the starter, grower, and overall phases, average daily feed intake was increased (quadratic; p<0.01) during the starter phase, and feed:gain ratio was decreased (quadratic; p<0.05) during the grower and overall phases. At the end of finisher phase, Met supplementation increased breast muscle content (quadratic; p<0.01) and thigh muscle content (linear; p<0.05), and decreased abdominal fat content (quadratic; p<0.02). Compared to the broiler fed DLM, broilers fed HMTBA had superior breast and thigh muscle coloration (p<0.01). Male broilers had higher weight gain and feed intake and better feed conversion than female broilers (p<0.01). The fat content of thigh muscle in female broilers was higher than that of male broilers (p<0.03). The best fit comparison of HMTBA vs. DLM was determined by Schwarz Bayesian Criteria index, which indicated that the average relative bioefficacy of HMTBA vs. DLM was 120% with 95% confidence limit 67 to 172%. These results indicated that Met supplementation improved growth performance and carcass quality of broilers fed corn-soybean-cottonseed meal diets irrespective of Met sources. Compared to DLM, HMTBA has the same molar bioefficacy on improving the growth performance and carcass quality of broilers; however, HMTBA fed birds had superior meat color to DLM fed birds.
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