Drying dynamics of meat highlighting areas of relevance to dry-aging of beef.

Drying of foods is a processing step, which has a variety of outcomes from improving shelf life and product stability, to reducing weight, or to achieving a targeted product eating quality. Drying is key step in the manufacturing of some dried meat products, such as jerky. It is also a major event that occurs when beef is dry-aged, where beef is exposed to air under defined conditions for an extended aging period. Although the conditions typically used to produce dried meat products are significantly different from those that prevail during dry-aging, both involve a gradual removal of water from muscle. As there is a paucity of research on the kinetics of the dehydration process occurring during dry-aging of beef, this paper comprehensively reviews models used to describe drying kinetics in other beef products, in order to gain insights regarding the key factors that impact water removal from meat. Consideration is given as to how the specific conditions during dry-aging such as air flows used (approximately 2 m/s), high air relative humidity, low temperature, and meat geometry will influence the kinetics of the drying. With regard to modeling, equations derived from Fick's second law of diffusion (e.g., thin-layer models) have been used to describe the drying kinetics of small-sized meat products. However, to apply Fick's law to dry-aging, some different considerations may need to be evaluated such as: tridimensional geometry (i.e., whole muscle); uniform initial moisture content; isotropic diffusion; negligible shrinkage;and a combination of internal and external resistances.

Transcriptome analysis of adipose tissue from pigs divergent in feed efficiency reveals alteration in gene networks related to adipose growth, lipid metabolism, extracellular matrix, and immune response.

Adipose tissue is hypothesized to play a vital role in regulation of feed efficiency (FE; efficiency in converting energy and nutrients into tissue), of which improvement will simultaneously reduce environmental impact and feed cost per pig. The objective of the present study was to sequence the subcutaneous adipose tissue transcriptome in FE-divergent pigs (n = 16) and identify relevant biological processes underpinning observed differences in FE. We previously demonstrated that high-FE pigs were associated with lower fatness when compared to their counterparts. Here, ontology analysis of a total of 209 annotated genes that were differentially expressed at a p < 0.01 revealed establishment of a dense extracellular matrix and inhibition of capillary formation as one underlying mechanism to achieve suppressed adipogenesis. Moreover, mechanisms ensuring an efficient utilization of lipids in high-FE pigs might be orchestrated by upstream regulators including CEBPA and EGF. Consequently, high-FE adipose tissue could exhibit more efficient cholesterol disposal, whilst inhibition of inflammatory and immune response in high-FE pigs may be an indicator of an optimally functioning adipose tissue. Taken together, adipose tissue growth, extracellular matrix formation, lipid metabolism and inflammatory and immune response are key biological events underpinning the differences in FE. Further investigations focusing on elucidating these processes would assist the animal production industry in optimizing strategies related to nutrient utilization and product quality.

RNA-seq of muscle from pigs divergent in feed efficiency and product quality identifies differences in immune response, growth, and macronutrient and connective tissue metabolism.

BACKGROUND: Feed efficiency (FE) is an indicator of efficiency in converting energy and nutrients from feed into a tissue that is of major environmental and economic significance. The molecular mechanisms contributing to differences in FE are not fully elucidated, therefore the objective of this study was to profile the porcine Longissimus thoracis et lumborum (LTL) muscle transcriptome, examine the product quality from pigs divergent in FE and investigate the functional networks underpinning the potential relationship between product quality and FE. RESULTS: RNA-Seq (n = 16) and product quality (n = 40) analysis were carried out in the LTL of pigs differing in FE status. A total of 272 annotated genes were differentially expressed with a P < 0.01. Functional annotation revealed a number of biological events related to immune response, growth, carbohydrate & lipid metabolism and connective tissue indicating that these might be the key mechanisms governing differences in FE. Five most significant bio-functions altered in FE groups were 'haematological system development & function', 'lymphoid tissue structure & development', 'tissue morphology', 'cellular movement' and 'immune cell trafficking'. Top significant canonical pathways represented among the differentially expressed genes included 'IL-8 signalling', 'leukocyte extravasation signalling, 'sphingosine-1-phosphate signalling', 'PKCθ signalling in T lymphocytes' and 'fMLP signalling in neutrophils'. A minor impairment in the quality of meat, in relation to texture and water-holding capacity, produced by high-FE pigs was observed. High-FE pigs also had reduced intramuscular fat content and improved nutritional profile in terms of fatty acid composition. CONCLUSIONS: Ontology analysis revealed enhanced activity of adaptive immunity and phagocytes in high-FE pigs suggesting more efficient conserving of resources, which can be utilised for other important biological processes. Shifts in carbohydrate conversion into glucose in FE-divergent muscle may underpin the divergent evolution of pH profile in meat from the FE-groups. Moreover, altered amino acid metabolism and increased mobilisation & flux of calcium may influence growth in FE-divergent muscle. Furthermore, decreased degradation of fibroblasts in FE-divergent muscle could impact on collagen turnover and alter tenderness of meat, whilst enhanced lipid degradation in high-FE pigs may potentially underlie a more efficient fat metabolism in these animals.

Use of a Screening Questionnaire to Identify Patients at Risk of Hyperglycemia Prior to Steroid Injection Therapy.

Objective: Patients with chronic pain considered for steroid injections are not consistently screened for undiagnosed or poorly controlled diabetes mellitus (DM) prior to steroid administration despite the known impact of corticosteroids on glucose control. A quality improvement project was undertaken to identify the prevalence of at-risk patients prior to steroid exposure through the development and implementation of a screening questionnaire. Subject: Three hundred and five adult patients with chronic pain who were being considered for an outpatient steroid injection were interviewed. Methods: Each patient was asked to complete an American Diabetes Association-validated diabetes mellitus screening questionnaire (ADAQ). In addition, patients responded to a set of nine questions that addressed common signs and symptoms of hyperglycemia. Results: The total prevalence of known DM in the study population was found to be 20% (60 patients), based on a documented history of diabetes mellitus. The ADAQ identified an additional 135 patients (45%) at high risk for undiagnosed DM. The full study questionnaire identified 143 patients (47.3%) who had no known history of diabetes mellitus as being at risk for hyperglycemia. In patients with known diabetes, five of nine symptom-based screening questions occurred with significantly frequency (P < 0.05): excessive thirst, excessive hunger, blurred vision, poor wound healing, and repeated infections. Conclusions: More than 60% of the patients screened had high risk of having inadequately treated DM. Thus, routine screening for DM should be considered in patients prior to treatment involving corticosteroids. The study questionnaire is a potentially cost-effective initial screening tool to determine which patients should undergo laboratory testing.

Use of a Risk-Stratification Tool in Identification of Potential Adrenal Suppression Preceding Steroid Injection Therapy in Chronic Pain Patients.

BACKGROUND: Patients who present for steroid injections are not routinely screened for potential hypothalamic-pituitary-adrenal (HPA) axis suppression from previous steroid exposure. Patients often receive various steroid therapies that are not reported by the patient or recorded in available medical records. Yet, HPA axis suppression has been reported with a single intra-articular injection. METHODS: An IRB-approved quality improvement questionnaire was implemented to comprehensively screen patients for risk of HPA axis suppression secondary to prior and/or concurrent corticosteroid use. This questionnaire was given to adult patients seen in a University Pain Management Clinic, who were being considered for a steroid injection, to define the extent of exposure to corticosteroids either by mouth, topically, inhaled, or systemic/local injection within the past 6 months. RESULTS: Two hundred patients completed the questionnaire. Eighty-nine patients (44.5%) screened positive for significant steroid exposure with a screen score of three or above. The average score for the screen positive group was 6.31 ± 3.47 (range 3-22). Women were 1.9 times more likely to screen positive than men (53.4% vs 27.5%, P < 0.0004). Otherwise, the screen positive and screen negative groups were similar in demographic characteristics (age, BMI, and diabetes status). CONCLUSIONS: Our results suggest that patients receive steroids from many sources and may be at risk for HPA axis suppression. Further testing is necessary to determine if these patients indeed have biochemical evidence of adrenal suppression. Utilization of a screening questionnaire might help identify patients who should be considered for HPA axis testing prior to steroid injections.

Modelling and numerical methods for identifying low-level adulteration in ground beef using near-infrared hyperspectral imaging (NIR-HSI).

Owing to the inherent characteristics of ground beef, adulteration presents a substantial risk for suppliers and consumers alike. This study developed a robust and novel method for identifying replacement fraud in ground beef with beef liver, beef heart, and pork using Near Infrared-Hyperspectral Imaging (NIR-HSI) coupled with chemometric and other statistical methods. More specifically, NIR-HSI provided an efficient and accurate means of identifying each type of adulteration using the classification model Genetic Algorithm (GA) - Backpropagation Artificial Neural Network (BPANN), showing perfect sensitivity and specificity (a value of 1.00) for the calibration and the validation sets for all types of adulteration. As an alternative to chemometric analysis, Hyperspectral Imaging-Root Mean Square (HSI-RMS) value, based on the RMScut-off calculation, was determined to discriminate types of adulterations without the need of resource-intensive modelling. This HSI-RMS approach provides a simple-to-use method that avoids the complexity of HSI data processing and aims to directly understand the similarity between different spectra of one sample in the pixel level. Different types of adulteration show noticeable differences reflected in the HSI-RMS value (varying from 55 to 1439), which demonstrate the potential of HSI-RMS concept as a novel and valuable alternative for assessing the HSI data and facilitating the identification of adulterants.

Incorporation of Sea Spaghetti (Himanthalia elongata) in Low-Salt Beef Patties: Effect on Sensory Profile and Consumer Hedonic and Emotional Response.

Seaweed is a naturally rich source of nutrients and exhibits techno-functional properties that are under study for their potential as ingredients in meat products. However, seaweed is associated with a particular flavor profile, and optimization of the sensory profile should be conducted alongside technical performance. This study investigated the feasibility of the application of sea spaghetti (Himanthalia elongata) in the production of low-salt beef patties and recorded the associated sensory profile and consumer hedonic-emotional response. Eight beef patty formulations with varying salt (0-1%) and seaweed (0-5%) contents were subjected to quantitative descriptive analysis via a trained sensory panel (n = 8) and six the formulations were selected for consumer testing (liking, emotional associations, saltiness perception, and purchase intent) by a group of 105 Irish resident consumers. The trained panel results showed that the intensity of seaweed odor, flavor, and visual presence in burgers was negatively related to the intensity of beef odor and flavor and that seaweed addition (5%) significantly increased the saltiness perception of low-salt burgers. Burgers with 1% added seaweed, although perceived by consumers as less salty, could substitute NaCl in low-salt beef patties without deterioration of their liking among regular burger consumers. Consumers associated all seaweed-containing samples, especially those containing 1% of sea spaghetti, with being good, pleasant, satisfied, and warm. The higher inclusion of sea spaghetti (2.5%) led to significantly lower overall liking and reduced purchase intent, while consumers associated this formulation with emotions such as being more adventurous, aggressive, and wild. Consumers who rejected seaweed burgers had the highest level of food neophobia and avoided foods with additives. The results demonstrate that 1% sea spaghetti seaweed can be successfully incorporated into low-salt beef patties, resulting in hedonic and emotional benefits without significantly increasing the salt content.

2D DIGE proteomic analysis of early post mortem muscle exudate highlights the importance of the stress response for improved water-holding capacity of fresh pork meat.

Variation in water-holding capacity (WHC), which presents a major economic burden to the swine industry, is considered to be underpinned by variation at a molecular and biochemical level. High-resolution 2D DIGE followed by MS analysis and Western blot were used to unravel the proteome of muscle exudate, collected following centrifugation, in the pH 4-7 range. A first 2DE-based protein map of this substrate was produced where 89 spots were successfully characterised. Two phenotypes divergent for WHC plus one intermediate were compared with a view to deciphering the biochemical processes impacting on variation in WHC. Twenty spots were observed to be altered across the phenotypes. Of these, 14 represented sixteen proteins including metabolic enzymes, stress response proteins and structural proteins. Triosephosphate isomerase and transferrin showed a major difference between the two extreme phenotypes, and may have potential as biological markers for WHC prediction. Several members of the HSPs family were highlighted. This proteomic study makes an important contribution towards a more detailed molecular view of the processes behind WHC and will provide a valuable resource for future investigations.

Transcriptome analysis of porcine M. semimembranosus divergent in intramuscular fat as a consequence of dietary protein restriction.

BACKGROUND: Intramuscular fat (IMF) content is positively correlated with aspects of pork palatability, including flavour, juiciness and overall acceptability. The ratio of energy to protein in the finishing diet of growing pigs can impact on IMF content with consequences for pork quality. The objective of this study was to compare gene expression profiles of Musculus semimembranosus (SM) of animals divergent for IMF as a consequence of protein dietary restriction in an isocaloric diet. The animal model was derived through the imposition of low or high protein diets during the finisher stage in Duroc gilts. RNA was extracted from post mortem SM tissue, processed and hybridised to Affymetrix porcine GeneChip® arrays. RESULTS: IMF content of SM muscle was increased on the low protein diet (3.60 ± 0.38% versus 1.92 ± 0.35%). Backfat depth was also greater in animals on the low protein diet, and average daily gain and feed conversion ratio were lower, but muscle depth, protein content and moisture content were not affected. A total of 542 annotated genes were differentially expressed (DE) between animals on low and high protein diets, with 351 down-regulated and 191 up-regulated on the low protein diet. Transcript differences were validated for a subset of DE genes by qPCR. Alterations in functions related to cell cycle, muscle growth, extracellular matrix organisation, collagen development, lipogenesis and lipolysis, were observed. Expression of adipokines including LEP, TNFα and HIF1α were increased and the hypoxic stress response was induced. Many of the identified transcriptomic responses have also been observed in genetic and fetal programming models of differential IMF accumulation, indicating they may be robust biological indicators of IMF content. CONCLUSION: An extensive perturbation of overall energy metabolism in muscle occurs in response to protein restriction. A low protein diet can modulate IMF content of the SM by altering gene pathways involved in lipid biosynthesis and degradation; however this nutritional challenge negatively impacts protein synthesis pathways, with potential consequences for growth.

Monitoring post mortem changes in porcine muscle through 2-D DIGE proteome analysis of Longissimus muscle exudate.

BACKGROUND: Meat quality is a complex trait influenced by a range of factors with post mortem biochemical processes highly influential in defining ultimate quality. High resolution two-dimensional DIfference Gel Electrophoresis (2-D DIGE) and Western blot were applied to study the influence of post mortem meat ageing on the proteome of pork muscle. Exudate collected from the muscle following centrifugation was analysed at three timepoints representing a seven day meat ageing period. RESULTS: The intensity of 136 spots varied significantly (p < 0.05) across this post mortem period and 40 spots were identified using mass spectrometry. The main functional categories represented were metabolic proteins, stress-related proteins, transport and structural proteins. Metabolic and structural proteins were generally observed to increase in abundance post mortem and many likely represent the accumulation of the degradation products of proteolytic enzyme activity. In contrast, stress-related proteins broadly decreased in abundance across the ageing period. Stress response proteins have protective roles in maintaining cellular integrity and a decline in their abundance over time may correlate with a reduction in cellular integrity and the onset of meat ageing. Since cellular conditions alter with muscle ageing, changes in solubility may also contribute to observed abundance profiles. CONCLUSIONS: Muscle exudate provided valuable information about the pathways and processes underlying the post mortem ageing period, highlighting the importance of post mortem modification of proteins and their interaction for the development of meat quality traits.

Addition of objective data to identify risk for medication misuse and abuse: the inconsistency score.

OBJECTIVE: To identify and quantify the rate of aberrant drug-taking behaviors using objective data. DESIGN: Institutional Review Board-approved anonymous, voluntary, quality improvement project. SETTING: University-based, multidisciplinary pain management center. SUBJECTS: Consecutive initial visit patients. METHODS: Patients were interviewed, asked to provide a urine sample, and filled out a brief questionnaire about recent prescription, over-the-counter, and illicit drug use. Discrepancies between patient report (PQ), the Virginia State prescription monitoring program (PMP), referring physician records (MRs), and the point-of-care urine drug screen (POC UDS) results were scored from 0 (none) to a maximum of 2 points (2+ discrepancies) for each potential comparator between data sets. Maximum potential inconsistency score (IS) was 16 points. RESULTS: Two hundred nine patients were interviewed to yield 118 specimens. Mean age of participants was 48.2 years (22-83 year); 65.3% were female. IS scores ranged from 1 to 11, and 52.5% of the patients had an IS ≥ 3. Higher IS scores correlated with higher numbers of pharmacies, prescribing physicians, prescriptions on the PMP, and presence of illicit substances in the urine. Addition of either POC UDS or PMP to PQ and MR increased identification of inconsistencies by >400%, and PMP plus UDS by >900%. CONCLUSIONS: Patient report and the medical record are inadequate to screen for aberrant drug-related behaviors. Addition of PMP and POC UDS contribute significantly to identification of inconsistencies through higher IS scores and differentiate patients at higher risk of medication misuse, abuse, or diversion. Comparison of multiple sources of objective information provides better insight into inconsistencies of report and behavior, and may assist in more appropriate and safer prescribing decisions.

DIGE Analysis of Animal Tissues.

Two-dimensional difference gel electrophoresis (2D-DIGE) is an acrylamide gel electrophoresis-based technique for protein separation and quantification in complex mixtures. The technique addresses some of the drawbacks of conventional 2D polyacrylamide gel electrophoresis (2D-PAGE), offering improved sensitivity, more limited experimental variation, and accurate within-gel matching. 2D-DIGE is based on direct labeling of proteins with isobaric fluorescent dyes (known as CyDyes: Cy2, Cy3, and Cy5) prior to isoelectric focusing (IEF). Here, up to two samples and a reference pool (internal standard) can be mixed and loaded onto IEF for first dimension prior to SDS (sodium dodecyl sulfate)-PAGE separation in the second dimension. After the electrophoretic run, the gel is imaged at the specific excitation wavelength for each dye, in sequence, and gel scans are recorded separately. For each individual protein spot, intensities recorded at the different wavelengths are integrated and the ratio between volumes normalized to that of the internal standard. This provides an immediate appreciation of protein amount variations under the different conditions tested. In addition, proteins of interest can still be excised and identified with conventional mass spectrometric techniques and further analyzed by other biochemical methods. In this chapter, we describe application of this methodology to separation and quantitation of protein mixtures from porcine muscle exudate, collected following centrifugation of muscle specimens (centrifugal drip) for the characterization of quality parameters of importance in meat industry.

Impact of sampling location and aging on the Longissimus thoracis et lumborum muscle proteome of dry-aged beef.

This study aimed to explore the differences in the proteome and molecular pathways between two sampling locations (external, internal) of bovine Longissimus thoracis et lumborum (LTL) muscles at 0, 21, and 28 days of dry-aging (i.e. 3, 24, and 31 days post-mortem). It further assessed the impact of post-mortem aging on the meat proteome changes and the biological processes at interplay. Proteins related to defence response to bacterium and regulation of viral entry into host cell were identified to be more abundant on the external location before dry-aging, which may be associated to the oxidative conditions and microbial activity to which post-mortem muscle is exposed during dressing, chilling, and/or quartering of the carcasses. This highlights the relevance of sampling from interior tissues when searching for meat quality biomarkers. As dry-aging progressed, the meat proteome and related biological processes changed differently between sampling locations; proteins related to cell-cell adhesion and ATP metabolic processes pathways were revealed in the external location at 21 and 28 days, respectively. On the other hand, the impact of aging on the proteome of the interior meat samples, evidenced that muscle contraction and structure together with energy metabolism were the major pathways driving dry-aging. Additionally, aging impacted other pathways in the interior tissues, such as regulation of calcium import, neutrophil activation, and regeneration. Overall, the differences in the proteome allowed discriminating the three dry-aging times, regardless of the sampling location. Several proteins were proposed for validation as robust biomarkers to monitor the aging process (tenderization) of dry-aged beef: TTN, GRM4, EEF1A1, LDB3, CILP2, TNNT3, GAPDH, SERPINI1, and OMD.

Molecular mechanisms contributing to the development of beef sensory texture and flavour traits and related biomarkers: Insights from early post-mortem muscle using label-free proteomics.

Beef sensory quality comprises a suite of traits, each of which manifests its ultimate phenotype through interaction of muscle physiology with environment, both in vivo and post-mortem. Understanding variability in meat quality remains a persistent challenge, but omics studies to uncover biological connections between natural variability in proteome and phenotype could provide validation for exploratory studies and offer new insights. Multivariate analysis of proteome and meat quality data from Longissimus thoracis et lumborum muscle samples taken early post-mortem from 34 Limousin-sired bulls was conducted. Using for the first-time label-free shotgun proteomics combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS), 85 proteins were found to be related with tenderness, chewiness, stringiness and flavour sensory traits. The putative biomarkers were classified in five interconnected biological pathways; i) muscle contraction, ii) energy metabolism, iii) heat shock proteins, iv) oxidative stress, v) regulation of cellular processes and binding. Among the proteins, PHKA1 and STBD1 correlated with all four traits, as did the GO biological process 'generation of precursor metabolites and energy'. Optimal regression models explained a high level (58-71%) of phenotypic variability with proteomic data for each quality trait. The results of this study propose several regression equations and biomarkers to explain the variability of multiple beef eating quality traits. Thanks to annotation and network analyses, they further suggest protein interactions and mechanisms underpinning the physiological processes regulating these key quality traits. SIGNIFICANCE: The proteomic profiles of animals with divergent quality profiles have been compared in numerous studies; however, a wide range of phenotypic variation is required to better understand the mechanisms underpinning the complex biological pathways correlated with beef quality and protein interactions. We used multivariate regression analyses and bioinformatics to analyse shotgun proteomics data to decipher the molecular signatures involved in beef texture and flavour variations with a focus on multiple quality traits. We developed multiple regression equations to explain beef texture and flavour. Additionally, potential candidate biomarkers correlated with multiple beef quality traits are suggested, which could have utility as indicators of beef overall sensory quality. This study explained the biological process responsible for determining key quality traits such as tenderness, chewiness, stringiness, and flavour in beef, which will provide support for future beef proteomics studies.

Addressing Clean Label Trends in Commercial Meat Processing: Strategies, Challenges and Insights from Consumer Perspectives.

BACKGROUND: The concept of a clean label is difficult to define, even in common language, as the interpretation of what a "clean" food is differs from one person to another and from one organisation to another. The lack of a unique definition and regulations of what the term "clean" means, along with the growing consumer demand for more "natural" and healthier foods, is posing new challenges for manufacturers and ingredient producers. The meat industry, in particular, has been affected by this new movement owing to negative attitudes and feelings consumers associate with consuming processed meat products. Scope and approach: The review scope is to describe attributes and associations around the "clean" label term by analysing the most recent ingredients, additives and processing methods currently available for meat manufacturers. Their application in meat, plant-based alternatives and hybrid meat/plant products, current limitations and challenges presented in consumer perception, safety and potential impacts on product quality are also presented. KEY FINDINGS AND CONCLUSIONS: The availability of a growing number of "clean" label ingredients provides a new suite of approaches that are available for application by meat processors to help overcome some of the negative connotations associated with processed meat products and also support plant-based meat alternatives and hybrids.

Influence of meat sample geometry on dehydration dynamics during dry-aging of beef.

Meat geometry may impact on the dehydration kinetics during dry-aging influencing the drying rate and, potentially, aspects of meat quality. In this study, three meat geometries (slices, steaks and sections) were prepared from three bovine Longissimuss thoracis et lumborum at 3 days post-mortem and were dry-aged at 2 °C, 75% relative humidity with an airflow of 0.5-2.0 m/s for 22 days (slices), 48 days (sections) and 49 days (steaks). Weights were recorded during dry-aging and drying curves were obtained for the three geometries, with the larger sections showing limited dehydration due to internal resistance to moisture migration from the core to the surface. Seven thin-layer equations were fitted to the dehydration data in order to model the drying kinetics during dry-aging. Thin-layer models described reliably the drying kinetics of the three geometries. In general, reduced k values (h-1) reflected the lower drying rates as thickness increased. The Midilli model provided the best fit for all geometries. Proximate analyses of the three geometries and bloomed colour of sections were measured at the start and the end of the dry-aging period. Moisture loss during dry-aging led to the concentration of protein, fat and ash contents; while no significant differences were found for L*, a* and b* values of sections before and after the dry-aging process. In addition, moisture content, water activity (aw) and LF-NMR measurements were taken at different locations within beef sections to further explore water dynamics during dry-aging.

SNP variation in the promoter of the PRKAG3 gene and association with meat quality traits in pig.

BACKGROUND: The PRKAG3 gene encodes the γ3 subunit of adenosine monophosphate activated protein kinase (AMPK), a protein that plays a key role in energy metabolism in skeletal muscle. Non-synonymous single nucleotide polymorphisms (SNPs) in this gene such as I199V are associated with important pork quality traits. The objective of this study was to investigate the relationship between gene expression of the PRKAG3 gene, SNP variation in the PRKAG3 promoter and meat quality phenotypes in pork. RESULTS: PRKAG3 gene expression was found to correlate with a number of traits relating to glycolytic potential (GP) and intramuscular fat (IMF) in three phenotypically diverse F1 crosses comprising of 31 Large White, 23 Duroc and 32 Pietrain sire breeds. The majority of associations were observed in the Large White cross. There was a significant association between genotype at the g.-311A>G locus and PRKAG3 gene expression in the Large White cross. In the same population, ten novel SNPs were identified within a 1.3 kb region spanning the promoter and from this three major haplotypes were inferred. Two tagging SNPs (g.-995A>G and g.-311A>G) characterised the haplotypes within the promoter region being studied. These two SNPs were subsequently genotyped in larger populations consisting of Large White (n = 98), Duroc (n = 99) and Pietrain (n = 98) purebreds. Four major haplotypes including promoter SNP's g.-995A>G and g.-311A>G and I199V were inferred. In the Large White breed, HAP1 was associated with IMF% in the M. longissmus thoracis et lumborum (LTL) and driploss%. HAP2 was associated with IMFL% GP-influenced traits pH at 24 hr in LTL (pHULT), pH at 45 min in LTL (pH(45)LT) and pH at 45 min in the M. semimembranosus muscle (pH(45)SM). HAP3 was associated with driploss%, pHULT pH(45)LT and b* Minolta. In the Duroc breed, associations were observed between HAP1 and driploss% and pHUSM. No associations were observed with the remaining haplotypes (HAP2, HAP3 and HAP4) in the Duroc breed. The Pietrain breed was monomorphic in the promoter region. The I199V locus was associated with several GP-influenced traits across all three breeds and IMF% in the Large White and Pietrain breed. No significant difference in promoter function was observed for the three main promoter haplotypes when tested in vitro. CONCLUSION: Gene expression levels of the porcine PRKAG3 are associated with meat quality phenotypes relating to glycolytic potential and IMF% in the Large White breed, while SNP variation in the promoter region of the gene is associated with PRKAG3 gene expression and meat quality phenotypes.

Variation in the IGF2 gene promoter region is associated with intramuscular fat content in porcine skeletal muscle.

Intramuscular fat (IMF) and subcutaneous fat (back fat-BF) are two of the major fat depots in livestock. A QTN located in the insulin-like growth factor 2 gene (IGF2) has been associated with a desirable reduction in BF depth in pigs. Given that the lipid metabolism of intramuscular adipocytes differs from that of subcutaneous fat adipocytes, this study aimed to search for genetic variation in the IGF2 gene that may be associated with IMF, as well as BF, in diverse pig breeds. Four proximal promoter regions of the IGF2 gene were characterised and the association of IGF2 genetic variation with IMF and BF was assessed. Six promoter SNPs were identified in four promoter regions (P1-P4; sequence coverage 945, 866, 784 and 864 bp, respectively) in phenotypically diverse F1 cross populations. Three promoter SNPs were subsequently genotyped in three pure breeds (Pietrain = 98, Duroc = 99 and Large White = 98). All three SNPs were >95% monomorphic in the Pietrain and Duroc breeds but minor alleles were at moderate frequencies in the Large White breed. These SNPs were linked and one was located in a putative transcription factor binding site. Five haplotypes were inferred and three combined diplotypes tested for association with IMF and BF in the Large White. As expected haplotype 1 (likely in LD with the beneficial QTN allele) was superior for BF level. In contrast, the heterozygote diplotype of the most common haplotypes (1 and 2) was associated with higher IMF and marbling scores compared to either homozygote. Gene expression analysis of divergent animals showed that IGF2 was 1.89 fold up-regulated in muscle with higher compared to lower IMF content. These findings suggest that genetic variation in the promoter region of the IGF2 gene is associated with IMF content in porcine skeletal muscle and that greater expression of the IGF2 gene is associated with higher IMF content.

Association between promoter polymorphisms in a key cytoskeletal gene (Ankyrin 1) and intramuscular fat and water-holding capacity in porcine muscle.

Ankyrin 1 (ANK1) is a positional and functional candidate gene for both bovine and porcine meat quality. The objectives of this study were: (1) to determine if the gene expression levels of ANK1 are associated with pig meat quality traits; (2) to examine polymorphisms in the promoter region of the porcine ANK1 gene for association with meat quality in diverse breeds; and (3) to search for putative transcription factor binding sites predicted to be altered by such polymorphisms. ANK1 gene expression was positively correlated with drip loss (%) in the Large White breed. Twelve novel SNPs were discovered in a 761 bp region of the ANK1 promoter in three phenotypically diverse F1 cross populations. Five were subsequently selected for association analysis with meat quality traits and genotyped in three pure pig breeds (Pietrain n = 98, Duroc n = 99 and Large White n = 98). Two of these five SNPs were associated with meat quality traits at the Bonferroni significance threshold: g.-606G>A with drip loss % in the Pietrain breed and g.-272G>A with intramuscular fat (IMF) in the LTL and SM. Following haplotype construction from SNP genotypes, Haplotype 3 was found to be associated with drip loss % at the Bonferroni level of significance in the Pietrain breed and Haplotype 5 was associated at the Bonferroni level with IMF in two muscles in the Large White breed. Further associations were observed at the nominal significance threshold. Our conclusion from this study is that SNPs in the ANK1 gene promoter could potentially contribute to genome-assisted selection SNP panels to improve IMF and water holding capacity on a breed basis.

Acceptable Inclusion Levels for Selected Brown and Red Irish Seaweed Species in Pork Sausages.

Commercially available Irish edible brown (Himanthalia elongata­sea spaghetti (SS), Alaria esculenta­Irish wakame (IW)) and red (Palmaria palmata­dulse (PP), Porphyra umbilicalis­nori) seaweeds were incorporated into pork sausages at 1%, 2.5%, and 5%. Proximate composition, salt, water-holding (WHC), cook loss, instrumental colour analysis, texture profile analysis (TPA), and sensory analysis were examined. Protein (13.14−15.60%), moisture (52.81−55.71%), and fat (18.79−20.02%) contents of fresh pork sausages were not influenced (p > 0.05) by seaweed type or addition level. The ash content of pork sausages containing PP, SS, and IW at 2.5% and 5%, and nori at 5%, were higher (p < 0.05) than the control sample. In comparison to the control, sausages containing nori, SS, and IW at 5% displayed higher (p < 0.05) WHC. Cook loss was unaffected (p > 0.05) by the addition of seaweeds into sausage formulations, compared to the control and within each seaweed. The addition of seaweeds into sausages had an impact on the surface colour (L* a* b*) and texture profile analysis (TPA) at different inclusion levels. Overall, hedonic sensory acceptability decreased (p < 0.05) in cooked sausages containing PP at 2.5% and 5%, and SS and IW at 5%.