Eating Time as a Genetic Indicator of Methane Emissions and Feed Efficiency in Australian Maternal Composite Sheep.

Previous studies have shown reduced enteric methane emissions (ME) and residual feed intake (RFI) through the application of genomic selection in ruminants. The objective of this study was to evaluate feeding behaviour traits as genetic indicators for ME and RFI in Australian Maternal Composite ewes using data from an automated feed intake facility. The feeding behaviour traits evaluated were the amount of time spent eating per day (eating time; ETD; min/day) and per visit (eating time per event; ETE; min/event), daily number of events (DNE), event feed intake (EFI; g/event) and eating rate (ER; g/min). Genotypes and phenotypes of 445 ewes at three different ages (post-weaning, hogget, and adult) were used to estimate the heritability of ME, RFI, and the feeding behaviour traits using univariate genomic best linear unbiased prediction models. Multivariate models were used to estimate the correlations between these traits and within each trait at different ages. The response to selection was evaluated for ME and RFI with direct selection models and indirect models with ETE as an indicator trait, as this behaviour trait was a promising indicator based on heritability and genetic correlations. Heritabilities were between 0.12 and 0.18 for ME and RFI, and between 0.29 and 0.47 for the eating behaviour traits. In our data, selecting for more efficient animals (low RFI) would lead to higher methane emissions per day and per kg of dry matter intake. Selecting for more ETE also improves feed efficiency but results in more methane per day and per kg dry matter intake. Based on our results, ETE could be evaluated as an indicator trait for ME and RFI under an index approach that allows simultaneous selection for improvement in emissions and feed efficiency. Selecting for ETE may have a tremendous impact on the industry, as it may be easier and cheaper to obtain than feed intake and ME data. As the data were collected using individual feeding units, the findings on this research should be validated under grazing conditions.

Lipid Oxidation and Colour Stability of Lamb and Yearling Meat (Muscle Longissimus Lumborum) from SheepSupplemented with Camelina-Based Diets after Short-,Medium-, and Long-Term Storage.

This study investigated the impact of feeding pelleted diets containing camelina (Camelina sativa L. Crantz) hay (CAHP) or camelina meal (CAMP) as a supplement compared with a control pellet (CONP) diet, without vitamin E fortification. The fatty acid profile, retail colour, and lipid oxidative stability of lamb and yearling meat (m. longissimus lumborum) stored for short-, medium-, or long-periods (2 days (fresh), 45 days and 90 days) under chilled to semi-frozen conditions were determined. The CAMP diet altered key fatty acids (p < 0.05) in a nutritionally beneficial manner for human health compared to the other diets, with increased total omega-3, decreased omega-6 fatty acids and decreased omega-6/omega-3 ratio of muscle. Muscle vitamin E concentration was lower (p < 0.05) for both camelina diets (CAMP and CAHP) when compared with the CONP diet, with the average concentrations less than 1 mg/kg muscle for all three treatments. Animal type and storage length were factors that all affected (p < 0.05) colour and lipid oxidative stability of meat. These results emphasise the importance of vitamin E concentration in meat stored for extended periods under semi-frozen conditions to maintain desirable meat colour during retail display, and to avoid off-flavour development of the cooked meat.

Expression quantitative trait loci in sheep liver and muscle contribute to variations in meat traits.

BACKGROUND: Variants that regulate transcription, such as expression quantitative trait loci (eQTL), have shown enrichment in genome-wide association studies (GWAS) for mammalian complex traits. However, no study has reported eQTL in sheep, although it is an important agricultural species for which many GWAS of complex meat traits have been conducted. Using RNA sequence data produced from liver and muscle from 149 sheep and imputed whole-genome single nucleotide polymorphisms (SNPs), our aim was to dissect the genetic architecture of the transcriptome by associating sheep genotypes with three major molecular phenotypes including gene expression (geQTL), exon expression (eeQTL) and RNA splicing (sQTL). We also examined these three types of eQTL for their enrichment in GWAS of multi-meat traits and fatty acid profiles. RESULTS: Whereas a relatively small number of molecular phenotypes were significantly heritable (h2 > 0, P < 0.05), their mean heritability ranged from 0.67 to 0.73 for liver and from 0.71 to 0.77 for muscle. Association analysis between molecular phenotypes and SNPs within ± 1 Mb identified many significant cis-eQTL (false discovery rate, FDR < 0.01). The median distance between the eQTL and transcription start sites (TSS) ranged from 68 to 153 kb across the three eQTL types. The number of common variants between geQTL, eeQTL and sQTL within each tissue, and the number of common variants between liver and muscle within each eQTL type were all significantly (P < 0.05) larger than expected by chance. The identified eQTL were significantly (P < 0.05) enriched in GWAS hits associated with 56 carcass traits and fatty acid profiles. For example, several geQTL in muscle mapped to the FAM184B gene, hundreds of sQTL in liver and muscle mapped to the CAST gene, and hundreds of sQTL in liver mapped to the C6 gene. These three genes are associated with body composition or fatty acid profiles. CONCLUSIONS: We detected a large number of significant eQTL and found that the overlap of variants between eQTL types and tissues was prevalent. Many eQTL were also QTL for meat traits. Our study fills a gap in the knowledge on the regulatory variants and their role in complex traits for the sheep model.

An alternative approach for sustainable sheep meat production: implications for food security.

BACKGROUND: A pelleted diet containing camelina hay (CAMH) or camelina meal (CAMM) as a supplement along with a control pellet (CONT) diet formulated with commonly available feeds during summer was used to investigate an alternative pathway for sustainable meat production. Sustainable meat production was based on a simple estimation of income from meat produced versus feed costs if animals were fed for an extended period over summer compared to early slaughter at the beginning of summer. Eighty maternal composite wether lambs (Composite) based on Coopworth genetics and 80 pure Merino wether yearlings were divided into 10 groups within breed (n = 8) using stratified randomisation based on liveweights. Following 1 week of adaptation to experimental diets, animals were fed experimental diets for up to10 weeks. RESULTS: Animals were slaughtered after either 8, 9 or 10 weeks of full feeding when the average liveweight of diet/genetic combination reached a weight appropriate for either 'heavy lamb' or 'heavy hogget' production, which occurred between 8 and 10 weeks of full feeding. There was no diet × breed interactions except for dressing percentage (DP), where Composite lambs fed the CAMH diet had the greatest DP (48.1 ± 0.35) and the Merino yearlings fed the CAMM diet the lowest DP (45.8 ± 0.33). Composite lambs gained 17.6-20.3 kg and Merino yearlings gained 10.7-12.9 kg liveweight. Based on their DP, this resulted in the production of approximately 8.3-9.5 kg additional carcass weight in Composites and 4.9-5.7 kg in Merinos, which in turn produced greater profit per Composite lamb and a small profit per Merino yearling. CONCLUSIONS: Composite lambs fed CAMM and CAMH had 5% greater carcass weights at slaughter compared to the CONT group, but dietary treatments did not change carcass weight of Merino yearlings at slaughter. The extended feeding approach offered the producer an estimated economic gain of AUD $20.00 to $25.00 when yearly average prices were used (Method 1) and AUD $40.00 to $47.70 when pre- and post-summer average prices were used (Method 2) per Composite lambs, but extra carcass gain did not result in the same profit per Merino yearling. Among the Composites, the profit for animals fed the CAMH and CAMM were AUD $2.75 to $4.50 greater than CONT group when full year average prices were applied while AUD $3.50 to $5.50 greater than CONT group when pre- and post-summer average prices were applied. However, we acknowledge a combination approach of extended feeding for a portion of animals already on ground and selling the remaining animals pre-summer with joining of additional ewes is the most likely strategy. Considering the scenario of extended feeding for 3 weeks, based on the growth rates observed for Composite lambs, if an additional 2 kg carcass weight per animal can be gained for 50% of the 22 million lambs slaughtered in Australia (= 11 million animals), it would potentially supply an additional 22 million kg of lamb carcasses produced per annum. This is equivalent to producing an extra 1 million lamb carcasses per annum weighing 22 kg each. Feeding Composite lambs for an extended period and selling Merino yearlings pre-summer may be a good option due to faster growth rate of Composites that may help quick turn-over to market.

Technical note: validation of an automated feeding system for measuring individual animal feed intake in sheep housed in groups.

The development of feeding systems that can individually measure and control feed intake in a group-housed environment would allow a greater understanding of sheep intake without compromising animal welfare and behavior through the removal of social interactions between sheep. This study validated an automated feeding system for measuring feed intake of individual sheep when housed in groups. Validation of the feeding system was conducted during three separate experiments. The validation sampling involved the activation of four individual "feed events," whereby four separate samples weighing approximately 50, 100, 200, and 400 g were removed from each feeder, with each feed event being linked to a specific radio frequency identification (RFID) tag. The feeder validation experiments evaluated the ability of the feeding system to 1) create a unique feed event every time a sample of pellets was collected from the feeder, 2) link the feed event to the correct RFID, and 3) accurately record the weight of feed that was manually removed. All feed events were initiated and logged in the feeding system with 100% of the events being linked to the correct test RFID. Concordance correlation coefficients between the feeding system-recorded feed weight and the manually removed weight were 0.99 within all three experiments. There was also no overall and little level-dependent bias between the weights measured by the feeding system and weights measured on the external scales. These results indicate the stability of the feeding system over time and consistency between the feeders within and across the three experiments. In conclusion, the automated feeding system developed for measuring individual animal feed intake was able to detect and record the unique electronic RFID associated with unique feed events and accurately capture the weight of feed removed. Furthermore, there was no change in the accuracy of the system from the start to the end of experimental periods, and the amount of feed removed in the feed event (or meal size) did not impact the accuracy of the results.

Understanding the impact of sire lean meat yield breeding value on carcass composition, meat quality, nutrient and mineral content of Australian lamb.

Advances in genomics and technology measuring body composition are now allowing sheep producers to select directly for increased lean meat yield (LMY) using Australian Sheep Breeding Values (ASBV). This experiment evaluated the impact of sire LMY ASBV on carcass composition, meat quality, nutrient and mineral content for lambs reared at pasture and finished in a feedlot. A 1% unit increase in sire LMY ASBV resulted in progeny that were leaner (0.8%) and had less fat (1.0%) on carcass. There was also a 0.2% reduction in the intramuscular fat content, a 3.2 N increase in meat toughness determined by shear force at day 5 ageing, a reduction in the redness of the fresh meat and a lower iron content. It is concluded that Australian sheep producers will need to incorporate ASBVs for other aspects of meat quality when selecting sires with increased LMY to avoid deterioration in meat quality, nutritional content of lamb and fresh meat colour.

Understanding the action of muscle iron concentration on dark cutting: An important aspect affecting consumer confidence of purchasing meat.

We investigated the association of muscle iron concentration, in addition to ultimate pH (pHU), on dark meat formation in sheep of different breeds fed forage-based diets. At 1 h simulated display, redness of meat (a*-value) increased (P < .0001) by about 3 units as the iron concentration increased from 10 to 22 mg/kg of meat, whereas the a*-value decreased by 2 units as pHU increased from 5.5 to 6.2 in fresh meat (P < .0001). After 90 days storage the corresponding responses were about 2 units increase for iron concentration and about 1 unit decrease for pHU, respectively. The results clearly show that increased muscle iron concentration was strongly associated with reduced dark cutting in fresh and stored meat evaluated at 1 h simulated display. We conclude that it may be desirable to measure iron concentration, along with pHU, for evaluation of the potential for carcasses to produce dark cutting meat, and for the meat to turn brown during display.

Development of VISNIR predictive regression models for ultimate pH, meat tenderness (shear force) and intramuscular fat content of Australian lamb.

This study investigated the effectiveness of visible-near-infrared (VISNIR) spectroscopy at classifying Australian lamb for: a) ultimate pH (pH 24), b) meat tenderness (i.e. shear force at day 5 of ageing, SF5) and c) intramuscular fat (IMF) content at 24 h post-slaughter using a custom-made handheld probe coupled with the ASD Labspec Pro instrument. VISNIR predictive regression models were developed. In the loin muscle (M. longissimus thoracis et lumborum), the models classified the predicted pH 24, SF5 and IMF content at above or below a threshold value with 94%, 98% and 88% accuracy, respectively. The observed difference between the actual and predicted value (i.e. the standard error of cross validation, SECV) for ultimate pH and IMF content are approaching accuracies required to attain highly reliable Meat Standards Australia grading standards. However, further development is required to improve the SECV for SF5.

Milk-derived ribonuclease 5 preparations induce myogenic differentiation in vitro and muscle growth in vivo.

Ribonuclease 5, also known as angiogenin, is a stable and abundant ribonuclease in milk whey protein, which is able to regulate several cellular functions, including capillary formation, neuron survival, and epithelial cell growth. Ribonuclease 5 is important for protein synthesis directly stimulating rRNA synthesis in the nucleolus. Here, we show that biologically active RNase5 can be purified from bovine milk. Furthermore, we show that milk-derived RNase5 directly stimulates muscle cell differentiation in vitro, inducing C2C12 cell differentiation and myogenesis. When supplemented into the diet of healthy adult mice, milk-derived RNase5 preparations promoted muscle weight gain and grip strength. Collectively, these data indicate that milk-derived RNase5 preparations exhibit a novel role in skeletal muscle cell function.

Peanut allergens alter intestinal barrier permeability and tight junction localisation in Caco-2 cell cultures.

BACKGROUND/AIMS: Allergen absorption by epithelia may play an important role in downstream immune responses. Transport mechanisms that can bypass Peyer's patches include transcellular and paracellular transport. The capacity of an allergen to cross via these means can modulate downstream processing of the allergen by the immune system. The aim of this study was to investigate allergen-epithelial interactions of peanut allergens with the human intestinal epithelium. METHODS: We achieved this using the human Caco-2 cell culture model, exposed to crude peanut extract. Western and immunofluorescence analysis were used to identify the cellular and molecular changes of peanut extract on the intestinal epithelium. RESULTS: Following exposure of Caco-2 cells to peanut extract, binding of the peanut allergens Ara h 1 and Ara h 2 to the apical cellular membrane and transcytosis across the monolayers were observed. Additionally, the co-localisation of the transmembrane tight junction proteins occludin, JAM-A and claudin-1, with the intracellular adhesion protein ZO-1 was modified. CONCLUSION: Disruption of Caco-2 barrier integrity through tight junction disruption may enable movement of peanut proteins across the intestinal epithelium. This accounts for peanut's increased allergenicity, compared to other food allergens, and provides an explanation for the potency of peanut allergens in immune response elicitation.

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.

Global survey of the bovine salivary proteome: integrating multidimensional prefractionation, targeted, and glycocapture strategies.

Saliva is easily obtainable from a large number of animals in a noninvasive manner and contains a wide diversity of compounds including hormones, metabolites, and proteins that may be a good source of biomarkers of health and disease. Here we have used a combination of multidimensional prefractionation, targeted, and glycocapture methodologies to profile the bovine salivary proteome. The nontargeted approach used four different separation methodologies consisting of SDS-PAGE, Off-gel fractionation, RP-HPLC, and SCX-HPLC. In the targeted approach, we've employed a hypothesis-based methodology by only selecting extracellular proteins from in silico data. Finally, the hydrazide capture methodology not only enabled us to identify formerly N-linked glycoproteins but it also provided a selective enrichment process for the identification of low abundance proteins. Together, the three different approaches identified 402 salivary proteins and 45 N-linked glycoproteins. A large number of these proteins have previously been uncharacterized in bovine saliva. To date, this is the largest global survey of the bovine salivary proteome and expands the potential of the diagnostic utility of this fluid to guide development of experiments seeking biomarkers for health traits (i.e., disease resistance) as well as feed conversion efficiency and productivity traits in dairy and beef cattle.

Problems associated with determining protein concentration: a comparison of techniques for protein estimations.

Although a range of methods are available for determining protein concentration, many scientists encounter problems when quantifying proteins in the laboratory. The most commonly used methods for determining protein concentration in a modern biochemistry laboratory would probably be the Lowry and/or the Bradford protein assays. Other techniques, including direct spectrophotometric analysis and densitometry of stained protein gels, are applied, but perhaps to a lesser extent. However, the reliability of all of the above techniques is questionable and dependent to some extent on the protein to be assayed. In this paper we describe problems we encountered when using some of the foregoing techniques to quantify the concentration of poly(adenosine diphosphate-ribose) polymerase-1 (PARP-1), a nuclear enzyme found in most eukaryotes. We also describe how, by using a fluorescence-based assay and amino acid analysis, we overcame the problems we encountered.