Gene expression identifies metabolic and functional differences between intramuscular and subcutaneous adipocytes in cattle.

BACKGROUND: This study used a genome-wide screen of gene expression to better understand the metabolic and functional differences between commercially valuable intramuscular fat (IMF) and commercially wasteful subcutaneous (SC) fat depots in Bos taurus beef cattle. RESULTS: We confirmed many findings previously made at the biochemical level and made new discoveries. The fundamental lipogenic machinery, such as ACACA and FASN encoding the rate limiting Acetyl CoA carboxylase and Fatty Acid synthase were expressed at 1.6-1.8 fold lower levels in IMF, consistent with previous findings. The FA elongation pathway including the rate limiting ELOVL6 was also coordinately downregulated in IMF compared to SC as expected. A 2-fold lower expression in IMF of ACSS2 encoding Acetyl Coenzyme A synthetase is consistent with utilisation of less acetate for lipogenesis in IMF compared to SC as previously determined using radioisotope incorporation. Reduced saturation of fat in the SC depot is reflected by 2.4 fold higher expression of the SCD gene encoding the Δ9 desaturase enzyme. Surprisingly, CH25H encoding the cholesterol 25 hydroxylase enzyme was ~ 36 fold upregulated in IMF compared to SC. Moreover, its expression in whole muscle tissue appears representative of the proportional representation of bovine marbling adipocytes. This suite of observations prompted quantification of a set of oxysterols (oxidised forms of cholesterol) in the plasma of 8 cattle exhibiting varying IMF. Using Liquid Chromatography-Mass Spectrometry (LC-MS) we found the levels of several oxysterols were significantly associated with multiple marbling measurements across the musculature, but (with just one exception) no other carcass phenotypes. CONCLUSIONS: These data build on our molecular understanding of ruminant fat depot biology and suggest oxysterols represent a promising circulating biomarker for cattle marbling.

Chinese Consumer Assessment of Australian Sheep Meat Using a Traditional Hotpot Cooking Method.

Hotpot is a widely popular cooking method for sheepmeat in China. This study measured the sensory responses of 720 untrained Chinese consumers to Australian sheepmeat cooked using a hotpot technique with methods based on Meat Standards Australia protocols. Shoulder and leg cuts of 108 lambs and 109 yearlings were scored on tenderness, juiciness, flavour and overall liking with linear mixed effects models used to analyse the influence of muscle type and animal factors on these scores. On average, shoulder cuts were more palatable than legs cuts for all sensory traits (p < 0.01) and lambs compared to yearlings (p < 0.05). Intramuscular fat and muscularity were identified as strong drivers of eating quality (p < 0.05), with greater palatability for both cuts as intramuscular fat increased (range 2.5 to 7.5%), and muscularity decreased (as measured through loin weight adjusted for hot carcase weight). Consumers were unable to detect differences between animal sire type and sex in sheepmeat hotpot. These findings suggest shoulder and leg cuts performed comparatively well in hotpot compared to previously tested sheepmeat cooking methods and emphasise the importance of balanced selection for quality and yield traits to ensure that consumer satisfaction is maintained.

Eating Quality of Australian Grass and Grain-Fed Lamb Equally Rated by US Consumers.

Anecdotal suggestions that US consumers perceive Australian sheepmeat as more "gamey" or "stale" compared to US sheepmeat are potentially attributable to the extended chilled shipping times contributing to longer-aged meat and predominately pasture-fed grazing systems. This study evaluated the impact of diet and extended storage times on Australian sheepmeat using sensory scores as assessed by US consumers. Meat samples from Australian sheep (n = 80) fed a grass or grain diet were aged in a vacuum at 1-2 °C for 5, 21 or 45 days. Untrained consumers (n = 960) at Texas Tech University (Lubbock, Texas) assessed samples for overall liking, tenderness, juiciness and flavour using a scale from 1 (worst) to 100 (best). In general, US consumers scored grain- and grass-fed samples within the same storage period similarly (p > 0.05). Furthermore, storage from 5 to 21 days improved sensory scores by a maximum of 28.6 for tenderness for grass-fed outside cuts (p < 0.05), while storage for 21 to 45 days did not improve eating quality for most cuts of both diets (p > 0.05). This is an interesting finding for the Australian sheepmeat industry as long storage time has no negative effect on eating quality and US consumers enjoyed grass- and grain-fed sheepmeat equally.

When balanced for precursor fatty acid supply echium oil is not superior to linseed oil in enriching lamb tissues with long-chain n-3 PUFA.

Vegetable oils containing stearidonic acid (SDA, 18 : 4n-3) are considered better precursors of long-chain n-3 PUFA (LC n-3 PUFA) than those with only α-linolenic acid (ALA, 18 : 3n-3). The present study re-examined this premise using treatments where added ALA from linseed oil was matched with ALA plus SDA from echium oil. Lambs (n 6) were abomasally infused with saline (control (C), 25 ml), echium oil low (EL, 25 ml), echium oil high (EH, 50 ml), linseed oil low (LL, 25 ml) or linseed oil high (LH, 50 ml) for 4 weeks. The basal ration used was identical across all treatments. EPA (20 : 5n-3) in meat increased from 6·5 mg in the C lambs to 16·8, 17·7, 13·5 and 11·7 (SEM 0·86) mg/100 g muscle in the EL, EH, LL and LH lambs, respectively. For muscle DPA (docosapentaenoic acid; 22 : 5n-3), the corresponding values were 14·3, 22·2, 18·6 18·2 and 19·4 (SEM 0·57) mg/100 g muscle. The DHA (22 : 6n-3) content of meat was 5·8 mg/100 g in the C lambs and ranged from 4·53 to 5·46 (SEM 0·27) mg/100 g muscle in the oil-infused groups. Total n-3 PUFA content of meat (including ALA and SDA) increased from 39 mg to 119, 129, 121 and 150 (SEM 12·3) mg/100 g muscle. We conclude that both oil types were effective in enhancing the EPA and DPA, but not DHA, content of meat. Furthermore, we conclude that, when balanced for precursor n-3 fatty acid supply, differences between linseed oil and echium oil in enriching meat with LC n-3 PUFA were of little, if any, nutritional significance.

Sire Breed, Litter Size, and Environment Influence Genetic Potential for Lamb Growth When Using Sire Breeding Values.

Lamb growth can be optimised with genetic selection using sire Australian sheep breeding values, however, breeding value expression has been shown to be reduced with poor nutrition. It was therefore hypothesised that the genetic potential for lamb growth would also be reduced, where production factors such as multiple births limit growth. Live weights at birth, weaning, and post-weaning were collected from more than 18,000 lambs produced over five years and eight locations of the Sheep Cooperative Research Centre Information Nucleus Flock experiment, and the impact of environment, production factors, and genotype was determined using mixed effects regression. The genetic potential for lamb growth was moderated by environment, multiple births, and sire type (p < 0.05). Twin lambs achieved 76% of the expected weight gain at weaning and 58% post-weaning. For triplet lambs weight gains were drastically less at approximately 30% of the expected gain at the same time points. Lambs born to maternal sires consistently had the poorest response to genetic selection, achieving approximately half the expected weight gain. Hence, producers need to temper expectations for growth based on genetic selection, or employ mitigation strategies such as precision feeding, the use of alternate breeds, or place emphasis on the genetic merit of other desirable traits.

Consumer Perceptions of Meat Redness Were Strongly Influenced by Storage and Display Times.

Lamb (n = 79) meat colour was scored by 879 untrained consumers using a scale of 0 (brown) to 100 (red). This consumer colour score (CCS) was obtained on m. longissimus lumborum (loin) and m. semimembranosus (topside), stored for short (5-7 days), medium (33-35 days), and long periods (110-112 days) and a retail display time of up to 4 days. Consumers perceived topside to be less red initially and changed from red to brown more rapidly when stored for the long-storage period (p < 0.01). Whereas, the initial CCS of loin samples were similar across the storage periods (p > 0.05). CCS and the instrument measure oxy/met (reflectance of light at wavelengths 630 nm and 580 nm) had a low correlation coefficient of 0.33 (p < 0.01). The propensity for lamb growth and leanness indicated by sire breeding values for lamb weight, eye muscle depth, eye muscle fat depth, and loin intramuscular fat had varied and inconsistent effects on CCS. Therefore, even the selection on CCS.

Lamb Age has Little Impact on Eating Quality.

There is an industry wide perception that new season lamb has better eating quality than old season lamb. This study aims to identify differences in consumer eating quality scores between two age classes in lamb. Consumer eating quality scores from eight cuts across the carcass were evaluated from new season (NS; n = 120; average age = 240 days) and old season lambs (OS; n = 121; average age = 328 days), sourced from four different flocks. Cuts were grilled (loin, topside, outside, knuckle and rump) or roasted (leg, shoulder, rack) and scored by untrained consumers for tenderness, juiciness, liking of flavour and overall liking. There was no difference in eating quality scores between the two age classes for the loin, leg, shoulder and rack. This was similarly shown in the topside with the exception of juiciness scores where NS lambs were higher than OS lambs. There was also a lack of age difference in the outside with the exception of flock 3 where NS lambs scored higher than OS lambs for all sensory traits. Across all sensory traits, OS lambs received on average 2.8 scores lower for the knuckle and 3.1 scores lower for the rump compared to NS lambs. These results show little difference in eating quality between NS and OS lamb, and highlight the potential to develop high quality OS or "autumn lamb" products, with a similar premium price at retail as NS lambs.

Influence of Demographic Factors on Sheepmeat Sensory Scores of American, Australian and Chinese Consumers.

Along with animal production factors, it is important to understand whether demographic factors influence untrained consumer perceptions of eating quality. This study examined the impact of demographic factors and sheepmeat consumption preferences on eating quality scores of American, Australian and Chinese untrained consumers. M. longissimus lumborum (LL) and m. semimembranosus (SM) were grilled according to sheep Meat Standards Australia protocols and evaluated by 2160 consumers for tenderness, juiciness, flavour and overall liking. Linear mixed effects models were used to analyse the impact of demographic factors and sheepmeat consumption habits on eating quality scores. Consumer age, gender, number of adults in a household and income had the strongest effect on sensory scores (P ≤ 0.05), although, the impact was often different across countries. Frequency of lamb consumption had an impact on sensory scores of American, Australian and Chinese consumers but larger sample sizes in some underrepresented subclasses for Australian and Chinese consumers are needed. Results suggest it is important to balance sensory panels for demographic factors of age, gender, number of adults and income to ensure sensory preferences are accurately represented for these particular populations.

The Impact of Pre-Slaughter Stress on Beef Eating Quality.

The study evaluated the relationship between pre-slaughter stress, plasma biomarkers and consumer-evaluated eating quality of pasture raised beef cattle (n = 488). The design tested steer only, heifer only and mixed sex cattle with a comparison of direct kill versus a 14 day rest period in abattoir holding paddocks prior to slaughter. Experiment One sourced cattle from four farms and tested shipping and road transport. Experiment Two sourced cattle from four farms and tested a commercial saleyard pathway. The impact on treatment on untrained consumer eating quality scores were tested on five muscle groups, m. psoas major, m. longissimus dorsi lumborum, m. biceps femoris, m. semitendinosis, and m. infraspinatus. Across all muscles, a two-week rest period had the biggest improvement in sensory score. Mixed groups scored lower in the outside muscle than non-mixed groups. However, the mixing response was inconsistent in the eye round muscle and not significant in the other muscles. Plasma glucose and L-lactate indicated a marked acute stress response at slaughter with a small detrimental impact on consumer score. The muscle damage enzyme markers creatine kinase (CK) and aspartate aminotransferase (AST) were strongly associated with a lower meat quality score (MQ4). Neither ß-hydroxybutyrate (ßHB) nor non-esterified fatty acids (NEFA) were associated with MQ4, suggesting that fat mobilisation does not impact consumer sensory score.

The Use of Biochemical Measurements to Identify Pre-Slaughter Stress in Pasture Finished Beef Cattle.

This study considered the relationship between pre-slaughter stressors and plasma biomarkers in 488 pasture-raised cattle across two experiments. The design aimed to test groups consisting of steer only, heifer only, and mixed sex cattle under direct kill versus rested (14 days in abattoir holding paddocks) protocols. In Experiment One, cattle were sourced from four farms, and transported by trucks and ships on the same day. In Experiment Two, cattle were sourced from four farms where a comparison was made between marketing via two commercial saleyards or direct farm gate consignment to abattoir. Blood samples were collected at exsanguination for subsequent analyses and relation to meat quality attributes. Muscle damage, as indicated by creatine kinase, is the biomarker most correlated to ultimate pH and muscle glycogen concentrations. A two-week rest period is effective for lowering this enzyme and improving muscle glycogen concentration. Although the cattle was subjected to a range of stress inducing treatments, we found that plasma biomarkers alone appeared insufficient for use as diagnostic stress indicators.

A supply chain approach to improving the shelf life of lamb meat; vitamin E concentration, electrical stimulation, ageing period and packaging system.

This study investigated the effects of vitamin E, electrical stimulation, aging and packaging system on the colour stability of lamb meat. Eighty crossbred wether lambs, 6-8months old were fed on either vitamin E or control pelleted diet for 31days prior to slaughter. Half of the carcases from each group were electrically stimulated before being split longitudinally into 2. Each side was randomly allocated 1 of the 4 aging periods (5days fresh, 10, 20 and 30days CO2). Muscle samples were set for retail display, after the respective aging period, colour was measured over 96h. Supplementing vitamin E nutritionally is likely to lengthen the shelf life of lamb products aged longer than 10days. Muscle vitamin E concentrations >3.0mg/kg are required to increase the shelf life of lamb cuts aged for up to 30days to 60h. Medium voltage electrical stimulation did not have a detrimental effect on the display life of aged lamb meat.

Genetic correlations between meat quality traits and growth and carcass traits in Merino sheep1.

Genetic correlations between 16 meat quality and nutritional value traits and live weight at various ages, live ultrasound fat and muscle depth, carcass measures, and carcass dissection traits were estimated for Merino sheep in the Information Nucleus (IN). Genetic correlations between live weight at various ages and the carcass traits are also reported. The IN comprised 8 genetically linked flocks managed across a range of Australian sheep environments. Meat quality traits included between 1,200 and 1,300 records for progeny from over 170 sires for intramuscular fat (IMF), lean meat yield (LMY), shear force (SF5), pH, meat color, and meat nutritional value traits including iron and zinc levels and long-chain omega-3 and omega-6 polyunsaturated fatty acid levels. The genetic correlations indicated that selection of Merino sheep to either reduce fat or increase muscle using ultrasound assessments will result in little change in IMF and SF5. Myoglobin levels would tend to be reduced following selection for reduced ultrasound fat depth (0.35 ± 0.21, 0.43 ± 0.14), whereas increases in myoglobin levels would occur due to selection for increased ultrasound muscle depth (0.25 ± 0.24, 0.38 ± 0.15). Selection for increased live weight will result in favorable correlated responses in hot carcass weight (0.76 to 0.97), dressing percentage (0.13 to 0.47), and carcass muscle (0.37 to 0.95), but unfavorable responses of increases in carcass fatness (0.13 to 0.65) and possible small reductions in muscle oxidative activity (-0.13 ± 0.14 to -0.73 ± 0.33) and iron content (-0.14 ± 0.15 to -0.38 ± 0.16), and a possible deterioration of shear force from selection at later ages (0.15 ± 0.26, 0.27 ± 0.24). Negligible changes are generally expected for LMY and meat color traits following selection for increased live weight (most genetic correlations less than 0.20 in size). Selection for increased LMY would tend to result in unfavorable changes in several aspects of meat quality, including reduced IMF (-0.27 ± 0.18), meat tenderness (0.53 ± 0.26), and meat redness (-0.69 ± 0.40), as well as reduced iron levels (-0.25 ± 0.22). These genetic correlations are a first step in assisting the development of breeding values for new traits to be incorporated into genetic evaluation programs to improve meat production from Merino sheep and other dual-purpose sheep breeds.

The effect of lamb carcase weight and GR depth on the production of value-added cuts - A short communication.

Times for the progressive breakdown of 95 lamb carcases were recorded to determine the impact of carcase weight and GR tissue depth on the time and therefore cost to produce value added retail cuts. Further analysis also assessed the potential to use these carcase traits as predictors of fabrication times. Regression modeling demonstrated there was a limited ability to predict the difference in time to fabricate mid value-added (R2=0.18) and extreme value-added (R2=0.12) cuts compared to traditional cuts, suggesting that other factors need to be considered. However, this study highlighted the significant increases in time required to fabricate more value-added cuts and to breakdown heavier carcases. Furthermore, this study demonstrated the changes to the saleable meat yield as the degree of fabrication increased, such that the average product prices increased ($20.64/kg for mid value added and $28.72/kg for extreme value added) compared to traditional retail cuts ($15/kg) to offset the increased labour of fabricating value-added cuts.

A GR/Impedance probe proves unsuitable for measuring GR depth in Australian lamb carcases.

Multiple studies were conducted at 3 abattoirs to determine the potential of a dual function GR/Impedance probe to measure GR soft- tissue depth (depth at the 12th rib, 110mm from the mid line of the carcase) of typical lamb carcases processed in Australia. Lamb carcases (1016) were measured approximately 25min post slaughter with the probe and a GR knife as the most commonly used measurement. Modelling indicated that variation in the measurement using the probe compared to the measurement using the knife occurred with increasing carcase weight and fatness as well as between different abattoirs. Consequently, the probe in its current design cannot provide the repeatable measurement required by industry.

Dietary Lecithin Supplementation Can Improve the Quality of the M. Longissimus thoracis.

Forty crossbred (Large White × Landrace × Duroc) female pigs (16.4 kg ± 0.94 kg) were used to investigate the effect of dietary lecithin supplementation on growth performance and pork quality. Pigs were randomly allocated to a commercial diet containing either 0, 3, 15 or 75 g lecithin/kg of feed during the grower and finisher growth phase. Pork from pigs consuming the diets containing 15 g and 75 g lecithin/kg had lower hardness ( P < 0.001) and chewiness ( P < 0.01) values compared to the controls. Dietary lecithin supplementation at 75 g/kg significantly increased ( P < 0.05) the linoleic acid and reduced ( P < 0.05) the myristic acid levels of pork compared to the control and the 3 g/kg and 15 g/kg lecithin supplemented treatments. Pigs fed the 75 g/kg lecithin supplemented diet had lower plasma cholesterol ( P < 0.05) at slaughter compared to pigs fed the control diet and the 3 g/kg and 15 g/kg lecithin supplemented treatments. These data indicate that dietary lecithin supplementation has the potential to improve the quality attributes of pork from female pigs.

Nutritional influences on some major enteric bacterial diseases of pig.

There are several enteric bacterial diseases and conditions of pigs that require control to prevent overt disease, to reduce morbidity and mortality, and to improve the efficiency of production. Traditionally, veterinarians, feed manufacturers and producers have relied upon antibiotics and minerals (for example, ZnO, CuSO4) in diets for a large part of this control. However, recent trends, particularly in Europe, are to reduce antimicrobial use and seek alternative or replacement strategies for controlling enteric bacterial diseases. The majority of these strategies rely on 'nutrition', taken in its broadest sense, to reduce the susceptibility of pigs to these diseases. Evidence to date suggests that specific dietary interventions, for example feeding very highly-digestible diets based on cooked white rice, can reduce the proliferation of a number of specific enteric bacterial infections, such as post-weaning colibacillosis. No simple and universal way to reduce susceptibility to pathogens in the gastrointestinal tract has been identified, and the underlying basis for many of the reported positive effects of 'nutrition' on controlling enteric infections lacks robust, scientific understanding. This makes it difficult to recommend dietary guidelines to prevent or reduce enteric bacterial diseases. Furthermore, some diseases, such as porcine intestinal spirochaetosis caused by Brachyspira pilosicoli, are sometimes associated with other pathogens (co-infections). In such cases, each pathogen might have different nutrient requirements, ecological niches and patterns of metabolism for which a variety of dietary interventions are needed to ameliorate the disease. Greater understanding of how 'nutrition' influences gut epithelial biology and immunobiology, and their interactions with both commensal and pathogenic bacteria, holds promise as a means of tackling enteric disease without antimicrobial agents. In addition, it is important to consider the overall system (i.e. management, housing, welfare) of pig production in the context of controlling enteric bacterial diseases.

Health beneficial long chain omega-3 fatty acid levels in Australian lamb managed under extensive finishing systems.

The variation in levels of the health claimable long chain omega-3 fatty acids, eicosapentaenoic acid (EPA, 20:5n-3) plus docosahexaenoic acid (DHA, 22:6n-3) across production regions of Australia was studied in 5726 lambs over 3 years completed in 87 slaughter groups. The median level of EPA plus DHA differed dramatically between locations and sometimes between slaughters from the same location. The ratio of EPA plus DHA from lambs with high values (97.5% quantile) to lambs with low values (2.5% quantile) also differed dramatically between locations, and between slaughters from the same location. Consistency between years, at a location, was less for the high to low value ratio of EPA plus DHA than for the median value of EPA plus DHA. To consistently obtain high levels of omega-3 fatty acids in Australian lamb, there must be a focus on lamb finishing diets which are likely to need a supply of α-linolenic acid (18:3n-3), the precursor for EPA and DHA.

An independent validation association study of carcass quality, shear force, intramuscular fat percentage and omega-3 polyunsaturated fatty acid content with gene markers in Australian lamb.

Previous association studies revealed several single nucleotide polymorphisms (SNPs) that explained the observed phenotypic variation for meat tenderness and long-chain omega-3 polyunsaturated fatty acid (PUFA) content of Australian lamb. To confirm the validity of these associated SNPs at predicting meat tenderness and omega-3 PUFA content, an independent validation study was designed. The OvineSNP50 genotypes of these animals were used to impute the 192 SNP Meat Quality Research (MQR) panel genotypes on nearly 6200 animals from the Cooperative Research Centre for Sheep Industry Innovation Information Nucleus Flock and Sheep Genomics Falkiner Memorial Field Station flock. Association analysis revealed numerous SNP from the 192 SNP MQR panel that were associated with carcass quality - fat depth at the C-site and eye muscle depth; shear force at day 1 and day 5 after slaughter (SF1 and SF5); and omega-3 PUFA content at P<0.01. However, 1 SNP was independently validated for SF5 (i.e. CAST_101781475). The magnitude of the effect of each significant SNP and the relative allele frequencies across Merino-, Maternal- and Terminal-sired progeny was determined. The independently validated SNP for SF5 and the associated SNP with omega-3 PUFA content will accelerate efforts to improve these phenotypic traits in Australian lamb.

Sources of variation of health claimable long chain omega-3 fatty acids in meat from Australian lamb slaughtered at similar weights.

The sources of variation of health claimable omega-3 polyunsaturated fatty acids (eicosapentaenoic acid, EPA+docosahexaenoic acid, DHA) in 2000 Australian lambs were investigated using 98 sires (Merino, maternal or terminal breeds) that were mated to about 5000 Merino or crossbred (Border Leicester×Merino) ewes. Pasture was supplemented with feedlot pellets, grains or hay as necessary, when the availability of quality green pasture was limited. Lambs were grown at 8 sites across Australia and when slaughtered the longissimus lumborum muscle was collected. Site and kills within sites were the major sources of variation for health claimable fatty acids. These environmental effects are likely to be driven by dietary background. The sire variance differed from about one twentieth to a half of the residual lamb within dam variation, depending on site and kill. This is the first comprehensive study to investigate on-farm sources of variation of long chain omega-3 polyunsaturated fatty acid content of lamb meat.