Chromosome-length genome assembly of Teladorsagia circumcincta - a globally important helminth parasite in livestock.

BACKGROUND: Gastrointestinal (GIT) helminthiasis is a global problem that affects livestock health, especially in small ruminants. One of the major helminth parasites of sheep and goats, Teladorsagia circumcincta, infects the abomasum and causes production losses, reductions in weight gain, diarrhoea and, in some cases, death in young animals. Control strategies have relied heavily on the use of anthelmintic medication but, unfortunately, T. circumcincta has developed resistance, as have many helminths. Vaccination offers a sustainable and practical solution, but there is no commercially available vaccine to prevent Teladorsagiosis. The discovery of new strategies for controlling T. circumcincta, such as novel vaccine targets and drug candidates, would be greatly accelerated by the availability of better quality, chromosome-length, genome assembly because it would allow the identification of key genetic determinants of the pathophysiology of infection and host-parasite interaction. The available draft genome assembly of T. circumcincta (GCA_002352805.1) is highly fragmented and thus impedes large-scale investigations of population and functional genomics. RESULTS: We have constructed a high-quality reference genome, with chromosome-length scaffolds, by purging alternative haplotypes from the existing draft genome assembly and scaffolding the result using chromosome conformation, capture-based, in situ Hi-C technique. The improved (Hi-C) assembly resulted in six chromosome-length scaffolds with length ranging from 66.6 Mbp to 49.6 Mbp, 35% fewer sequences and reduction in size. Substantial improvements were also achieved in both the values for N50 (57.1 Mbp) and L50 (5 Mbp). A higher and comparable level of genome and proteome completeness was achieved for Hi-C assembly on BUSCO parameters. The Hi-C assembly had a greater synteny and number of orthologs with a closely related nematode, Haemonchus contortus. CONCLUSION: This improved genomic resource is suitable as a foundation for the identification of potential targets for vaccine and drug development.

Contribution of the Immune Response in the Ileum to the Development of Diarrhoea caused by Helminth Infection: Studies with the Sheep Model.

Gastrointestinal helminths are a global health issue, for humans as well as domestic animals. Most studies focus on the tissues that are infected with the parasite, but here we studied the ileum, a tissue that is rarely infected by helminths. We tested whether inflammation in the ileum contributes to the development and severity of diarrhoea, by comparing sheep that are susceptible (n = 4) or resistant (n = 4) to the disease. We analyzed the ileum transcriptome using RNASeq sequencing approach and various bioinformatics tools including FastQC, STAR, featureCounts, DESeq2, DAVID, clusterProfiler, Cytoscape (ClusterONE) and EnrichR. We identified 243 differentially expressed genes (DEGs), of which 118 were up-regulated and 125 were down-regulated DEGs in the diarrhoea-susceptible animals compared to the diarrhoea-resistant animals. The resulting DEGs were functionally enriched for biological processes, pathways and gene set enrichment analysis. The up-regulated DEGs suggested that an inflammatory immune response was coupled with genes involved in 'Th2 immune response' and 'anti-inflammatory response'. The down-regulated DEGs were related to ion transport, muscle contraction and pathways preventing inflammation. We conclude that i) susceptibility to helminth-induced diarrhoea involves an inflammatory response at a non-infectious site; ii) down-regulation of pathways preventing inflammation can contribute to the severity of diarrhoea; and iii) genes involved in anti-inflammatory responses can reduce the inflammation and diarrhoea.

Mineral profiling of muscle and hepatic tissues of Australian Merino, Damara and Dorper lambs: Effect of weight loss.

Seasonal weight loss (SWL) is a major constraint to extensive animal production systems. The Australian sheep production is based on merino sheep, a European breed not tolerant to SWL. Tolerant alternative breeds such as the fat-tailed Damara and the Dorper have been increasingly used in Australia and elsewhere, due to their robustness. The aim of this study was to understand the mineral profile of muscle and liver tissues of Australian Merino, Damara and Dorper, when subjected to SWL in order to understand SWL-tolerance physiology. Twenty-four lambs were divided randomly between growing (control) and nutritionally restricted groups for each breed. The trial lasted 42 days. Animals were weighed bi-weekly and at the end of the trial, lambs were slaughtered. Liver and muscle samples were taken immediately after slaughter. Mineral assessment was carried out using inductively coupled plasma-optical emission spectrometry. Analysis of variance showed mineral concentrations were generally increased in the muscle of restricted animals, mainly because of fat tissue mobilization. An increase in Zn and Fe concentrations indicates an increase of enzymatic activity in the liver of restricted sheep as well as differential abundance of Fe-containing proteins. High concentrations of Cu in the liver of Dorper indicate higher ability to accumulate this element, even under SWL.

Amino acid profiles of muscle and liver tissues of Australian Merino, Damara and Dorper lambs under restricted feeding.

Seasonal weight loss (SWL) is a major constraint in extensive animal production systems in the tropics and Mediterranean. The objective of this study was to characterize the amino acid profile of muscle and hepatic tissues of Australian Merino, Damara and Dorper lambs under restricted feeding to evaluate the impact of SWL at the metabolic and physiological levels. SWL induced generalized muscle protein breakdown among restricted groups of all breeds, with varying intensity. Dorper breed mobilized less muscle amino acids when under these conditions, with the Damara having frequent significant differences, namely by having lower amino acid concentrations in the muscle of restricted lambs. Damara lambs showed greater ability to catabolize branched-chain amino acids in the muscle tissue, which indicates yet another mechanism that provides the Damara with the necessary tools to endure harsh conditions. Overall, the Damara breed mobilized more muscle amino acids than the other breeds, with a better capacity to catabolize branched-chain amino acids in the muscle, while maintaining muscle structural integrity.

Revealing the associated microflora hosted by the globally significant parasite Trichostrongylus colubriformis.

Trichostrongylus colubriformis is a parasitic helminth that primarily infects small ruminants, causing substantial economic losses in the livestock industry. Exploring the microbiome of this helminth might provide insights into the potential influence of its microbial community on the parasite's survival. We characterised the intestinal microbiome of T. colubriformis that had been collected from the duodenum of sheep, and compared the helminth microbiome with the duodenal microbiome of its host, aiming to identify contributions from the helminth's environment. At the same time, we explored the isolation of fastidious organisms from the harvested helminth. Primary alpha and beta diversity analyses of bacterial species revealed statistically significant differences between the parasite and the host, in terms of species richness and ecological composition. 16S rRNA differential abundance analysis showed that Mycoplasmoides and Stenotrophomonas were significantly present in T. colubriformis but not in the duodenal microbiome of the sheep. Furthermore, two bacteria, Aeromonas caviae and Aeromonas hydrophila, were isolated from T. colubriformis. Examinations of the genome highlight differences in genome size and profiles of antimicrobial resistance genes. Our results suggest that T. colubriformis carries a specific bacterial community that could be supporting the helminth's long-term survival in the host's digestive system.

Utilising male stimulus to improve the reproductive efficiency of 8-month-old nulliparous ewes and adult parous ewes.

We tested whether utilising the male effect to stimulate ewes before the mating period can reduce the time to conception following the introduction of entire rams, and increase fertility, prolificacy, and reproductive rate (number of fetuses per 100 ewes exposed to fertile rams). A retrospective analysis was used to analyse records from 59,716 ewes collected over 34 years (1986-2020) from seven genotypes: Border Leicester, Composite (crossbred), Dorset, Merino, Dorset x Polypay, Rambouillet, White Suffolk. The dataset also included nulliparous young ewes (mated at age 8 months) and adult parous ewes. Vasectomized rams were used to stimulate 20,632 ewes before a mating period that lasted 2 or 3 estrous cycles, and the outcomes were compared with those from 39,084 ewes that had not been stimulated. Independently of genotype, utilising the male stimulus advanced the average conception date by 8 days for young ewes (P < 0.0001) and by 1 day for adult ewes (P < 0.0001). The male stimulus also increased the proportion of ewes that conceived in their first cycle by 33 % for young ewes and by 6 % for adult ewes (P < 0.0001). For the cycle of conception, there were significant (P < 0.0001) effects of two interactions: male stimulus x age at mating and male stimulus x live weight at mating. The male stimulus improved fertility in both adult ewes (99.8 % vs 89 %; P < 0.001) and young ewes (77.7 % vs 81.3 %; P < 0.001). The male stimulus increased the number of young ewes (41.9 % vs 11.1 %; P < 0.001) and adult ewes (16.6 % vs 2.7 %; P < 0.001) that conceived multiple fetuses in the first 17 days of the mating period. The reproductive rate was improved by the male stimulus in young ewes (129 % vs 135 %; P < 0.001) but not in adult ewes (120 % vs 122 %; P = 0.12). When all animals for all breeds were included in the analyses, there were improvements in fertility, prolificacy, and reproductive rate as age and live weight increased at mating (P < 0.0001). We conclude that, independently of genotype, utilising the male stimulus before the mating period reduces the time to conception and improves reproductive performance in both young and adult ewes.

Assessing carcass and meat characteristics of Damara, Dorper and Australian Merino lambs under restricted feeding.

Seasonal weight loss (SWL) is the most pressing constraint in ruminant production systems in tropical climates. SWL is controlled using supplementation, which is costly and difficult to implement in extensive systems, or using breeds adapted to tropical hot dry climates, like the Damara and Dorper. Albeit 15 years in Australia, little is known on how these sheep compare to Australian Merino. Here, the responses of the Damara, Dorper and Merino breeds to nutritional stress were compared. Seventy-two 6-month-old ram lambs, 24 from each breed, were allocated to growth (gaining 100 g/day) or restricted diets (losing 100 g/day, 85% of maintenance needs). Animals were weighed and carcass and meat characteristics determined. Results point out to the existence of important differences between the three genotypes, in particular between the Merino and the Southern African breeds. Additionally, Merino ram lambs seem to have been more influenced by SWL than the other two, with consequences on meat characteristics.

Targeted lipidomics coupled with machine learning for authenticating the provenance of chicken eggs.

Free-range eggs are ethically desirable but as with all high-value commercial products, the establishment of provenance can be problematic. Here, we compared a simple one-step isopropanol method to a two-step methyl-tert-butyl ether method for extracting lipid species in chicken egg yolks before liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. The isopropanol method extracted 937 lipid species from 20 major lipid subclasses with high reproducibility (CV < 30 %). Machine learning techniques could differentiate conventional cage, barn, and free-range eggs using an external test dataset with an accuracy of 0.94, 0.82, and 0.82, respectively. Lipid species that differentiated cage eggs were predominantly phosphocholines and phosphoethanolamines whilst the free-range egg lipidomes were dominated by acylglycerides with up to three fatty acids. The lipid profiles were found to be characteristic of the cage, barns, and free-range eggs. The lipidomic analysis together with the statistical modeling approach thus provides an efficient tool for verifying the provenance of conventional chicken eggs.

Investigating the development of diarrhoea through gene expression analysis in sheep genetically resistant to gastrointestinal helminth infection.

Gastrointestinal helminths infect livestock causing health problems including severe diarrhoea. To explore the underlying biological mechanisms relating to development and control of diarrhoea, we compared 4 sheep that were susceptible to development of diarrhoea with 4 sheep that were diarrhoea-resistant. Transcriptomes in the tissues where the parasites were located were analyzed using RNASeq. By considering low-diarrhoea sheep as control, we identified 114 genes that were down-regulated and 552 genes that were up-regulated genes in the high-diarrhoea phenotype. Functional analysis of DEGs and PPI sub-network analysis showed that down-regulated genes in the high-diarrhoea phenotype were linked to biological processes and pathways that include suppression of 'antigen processing and presentation', 'immune response', and a list of biological functional terms related to 'suppression in immune tolerance'. On the other hand, up-regulated genes in the high-diarrhoea phenotype probably contribute to repair processes associated with tissue damage, including 'extracellular matrix organization', 'collagen fibril organization', 'tissue morphogenesis', 'circulatory system development', 'morphogenesis of an epithelium', and 'focal adhesion'. The genes with important roles in the responses to helminth infection could be targeted in breeding programs to prevent diarrhoea.

Bacterial communities in the gastrointestinal tract segments of helminth-resistant and helminth-susceptible sheep.

BACKGROUND: Helminth parasitism is a world-wide problem in livestock industries, with major impacts on health, welfare and productivity. The role of the gut microbiota in host-helminth interactions in ruminants has been extensively examined and the present study added to this body of knowledge by assessing the effects of resistance and susceptibility to helminth infection in the gastro-intestinal tract (GIT). Australian Sheep Breeding Values (ASBVs) for faecal egg count (FEC) were used to select the 10 highly helminth-susceptible (High-FEC) and 10 highly helminth-resistant (Low-FEC) sheep. FEC status was confirmed during the experiment. Using samples from the faeces and the lumen of the rumen, abomasum, duodenum, jejunum, ileum, caecum, and colon, DNA was extracted and used for 16 rRNA gene amplicon sequencing. RESULTS: The most frequent genera identified along the GIT were Eubacterium, Oscillibacter, and Ruminococcus. Intersectoral-specialization zones were identified along the GIT, with the duodenum displaying major differences between the High-FEC and Low-FEC animals in values for alpha and beta diversity. After taking all samples into account and adjusting for GIT segment, the High-FEC and Low-FEC sheep differed significantly for four genera Butyrivibrio, Mycoplasma, Lachnoclostridium and Succiniclasticum. In the duodenum, the abundances of Aminipila, Lachnoclostridium and Mogibacterium differed significantly between the High-FEC and Low-FEC sheep. In the ileum, on the other hand, the genus Mycoplasma was significantly depleted in the Low-FEC group. CONCLUSIONS: The gastro-intestinal microbial profile varies widely between helminth-resistant and helminth-susceptible sheep. Each GIT section appears to support a particular bacterial composition leading to inter-sectoral differences among the various microbial communities. The microbial populations were most rich and diverse in the duodenum of helminth-resistant sheep, comprising bacterial genera that generally ferment carbohydrates. This observation suggests that helminth-resistant sheep can reorganize the duodenal microbiome taxa which may restrict the development of parasites.

A note on the reproductive performance of Damara, Dorper and Merino sheep under optimum management and nutrition for Merino ewes in the eastern wheatbelt of Western Australia.

The reproductive performance of 48 Damara, 42 Dorper and 46 Merino ewes was evaluated under an optimum nutritional regime for Merino ewes that included one annual joining in a mixed (cropping and sheep) farming system in the eastern wheatbelt of Western Australia (W.A.) over a 3-year period. In 2005, when the Damara, Dorper and Merino ewes were aged between 8 and 9 months at joining and weighed 41.2, 42.4 and 33.3 kg with average body condition scores of 2.3, 2.4 and 1.8, respectively, their weaning rates were 71% (Damara), 81% (Dorper) and 13% (Merino). The Merino ewes had significantly lighter body weights and were lower conditioned (p < 0.001) resulting in significantly lower weaning rates (p < 0.001). In 2006, the Damara, Dorper and Merino ewes weighed 61.9, 69.5 and 61.6 kg at joining, with body condition scores of 2.8, 3.3 and 2.8, respectively, resulting in weaning rates of 52% (Damara), 122% (Dorper) and 117% (Merino). While the Damara and Merino weights and body condition scores were not different (p > 0.05), the Damara weaning rates were significantly lower compared with the Dorper and Merino groups (p < 0.001). The Dorpers were heavier and greater conditioned (p < 0.001); however, the weaning rates between the Dorper and Merino were not different (p > 0.05). In 2007, the Damara, Dorper and Merino ewes weighed 71.6, 77.1 and 70.2 kg at joining with body condition scores of 2.8, 2.8 and 2.5, respectively. Again, the Dorper ewes were heavier (p < 0.001) than the Damaras and Merinos, which were not different (p > 0.05). The Damara and Dorper had the same body condition while the Merino ewes were less and different (p < 0.05). However, there were differences in weaning rates between the three breeds (p < 0.001), of 64% (Damara), 105% (Dorper) and 134% (Merino). Over the 3-year period, under a nutritional management system optimal for Merino sheep and with one annual joining, with increasing age the Merino ewes weaned more lambs. However, while the Damara and Dorper ewes conceived and weaned relatively high lamb numbers when they were joined as lambs, their reproductive performance decreased over time. This nutritional regime resulted in increased levels of fatness of the Damara and Dorper ewes with no increase in reproduction rates under an annual joining system. A significant factor for the lower conception and weaning rates in the Damara ewes was the enlarged fat tail due to the increased fat levels, which made it difficult for the rams to impregnate the ewes.

Ovine liver proteome: Assessing mechanisms of seasonal weight loss tolerance between Merino and Damara sheep.

The effect of feed restriction on the liver protein profiles of two different breeds of sheep was studied. We compared Merino with the Damara, breeds with respectively low and high tolerance to nutritional stress. Each breed was grouped into two nutritional treatments: restricted (12-14% loss of live weight) and control (maintenance). The trial lasted 42 days. Animals were sacrificed and liver samples subjected to label free shotgun proteomics. The resultant proteins had both their fold change and statistical significance in an unpaired t-test calculated to identify differential protein abundance. The tool WebGestalt was utilized to perform an Overrepresentation Enrichment Analysis (ORA) for gene ontology terms associated with the significant proteins. We further validated shotgun proteomics findings using a selected reaction monitoring (SRM)-based targeted proteomics approach, where similar trends in regulation were obtained for a subset of relevant proteins across an independent cohort of animals. Results confirm that Damara has adapted to nutritional stress by mobilizing stored fatty acids within adipose tissue and converting them to energy more efficiently than Merino. Finally, Merino had an overabundance pattern primarily directed to protein synthesis pathways. Regulated proteins identified may be used as a basis for marker selection towards tolerance to nutritional stress. BIOLOGICAL SIGNIFICANCE: Sheep are one of the most important livestock animals. They remain however poorly understudied and described. Seasonal weight loss (SWL) due to pasture scarcity during the dry season is one of the most limiting conditions to ruminant production in the tropics. To counter SWL, farmers may use supplementation, expensive or difficult to implement. A more suitable long-term solution would be to use breeds that are naturally adapted to nutritional stress. This work contrasts two breeds with different levels of tolerance to SWL, the Damara and the Merino, respectively well and poorly adapted. Comparison is conducted at the level of the hepatic tissue and using label free proteomics. This work identifies a series of pathways in the liver of the Damara via label free proteomics that suggest a unique fatty acid metabolic process within this breed. Proteins that have increased abundance in the Damara in association with fatty acid metabolism may be used as potential markers of tolerance to nutritional stress. This research will pave the way for more viable, long-term solutions for farmers facing annual production problems due to drought in the tropics and the Mediterranean region.

The ovine hepatic mitochondrial proteome: Understanding seasonal weight loss tolerance in two distinct breeds.

Seasonal weight loss (SWL) is a primary constraint for farmers in the Mediterranean and tropics. One cost-effective solution to SWL is utilizing breeds like the Damara sheep that have adapted to deal with nutritional stress. Previous studies concluded that one of the adaptation mechanisms of SWL is a specialized fatty acid metabolism. Accordingly, hepatic-mitochondrial proteomes were compared across two different breeds (24 sheep total, Merino, n = 12 and Damara, n = 12) and two different diets (restricted vs unrestricted diet, 6 per breed, per diet, 24 total). Mitochondrial-proteins were isolated and relatively quantified using Blue native PAGE / 2D-electrophoresis and then analyzed via mass spectrometry. The tool ReviGO summarized the proteomes' gene-ontology terms. A total of 50 proteins were identified with 7 changing significantly in abundance (ANOVA p-value<0.05). Specific abundance patterns of corticosteroid and inflammatory response-associated proteins such as annexin and glutamate dehydrogenase suggests that the Damara has an unusual inflammation response when subjected to SWL in addition to its unique metabolism. All significant proteins warrant further study; Annexin in particular shows promise as a potentially useful biomarker.

Behavior and Electrophysiological Response of Gravid and Non-Gravid Lucilia cuprina (Diptera: Calliphoridae) to Carrion-Associated Compounds.

The Australian blow fly, Lucilia cuprina Wiedmann (Diptera: Calliphoridae), is a major cause of myiasis (flystrike) in Merino sheep in Australia and New Zealand and, as a primary colonizer of fresh carrion, also an important species in forensic investigations. Olfaction is considered the most important cue for insects to rapidly locate carrion over long distances, so the first carrion visitors are predicted to be very sensitive to carrion-related volatile compounds. We studied the responses of the Australian blow fly, Lucilia cuprina, to the carrion-associated compounds dimethyl trisulfide (DMTS), butyric acid, 1-octen-3-ol and indole. We also tested 2-mercaptoethanol, a compound commonly used in fly traps in Australia. We investigated whether responses of the flies are affected by their ovarian status by comparing responses of gravid and non-gravid L. cuprina in electroantennography (EAG) and two-choice laboratory bioassays. All four compounds evoked an EAG response, while only DMTS evoked responses in gas chromatography-mass spectrometry electroantennographic detection (GCMS-EAD) analyses and two-choice bioassays. Gravid flies detected lower doses of the test compounds than non-gravid flies. Our results indicate that DMTS is an important semiochemical for L. cuprina to locate carrion resources, and has potential for use in fly traps for flystrike control. Our observations also suggest that the greater sensitivity of gravid L. cuprina allows them to find fresh carrion quickly to maximize reproductive success by avoiding unsuitable degraded carrion.

The sheep (Ovis aries) muscle proteome: Decoding the mechanisms of tolerance to Seasonal Weight Loss using label-free proteomics.

Seasonal Weight Loss (SWL) is one of the most pressing issues in animal production in the tropics and Mediterranean. This work aims to characterize muscle proteome changes as a consequence of SWL in meat producing sheep, using a label-free proteomics approach. We compare three breeds: the Australian Merino (SWL susceptible), the Damara (SWL tolerant) and the Dorper (SWL intermediate tolerance). We identified 668 proteins of the sheep proteome, 95 with differential regulation. Also we observe that the more vulnerable to SWL a breed is, the more differential abundance proteins we find. Protein binding was the most frequently altered molecular function identified. We suggest 6 putative markers for restricted nutritional conditions independently of breed: ferritin heavy-chain; immunoglobulin V lambda chain; transgelin; fatty acid synthase; glutathione S-transferase A2; dihydrodiol dehydrogenase 3-like. Moreover, we suggest as related to SWL tolerance: S100-A10 Serpin A3-5-like and Catalase, subject however to necessary validation assays. The identification of SWL-tolerance related proteins using proteomics will lead to increased stock productivity of relevant interest to animal production, particularly if identified at the muscle level, the tissue of economic importance in meat production. BIOLOGICAL SIGNIFICANCE: Seasonal Weight Loss (SWL) is the most pressing issue in animal production in the tropics and the Mediterranean. To counter SWL, farmers often use animal breeds that have a natural ability to withstand pasture scarcity. Here we study the sheep muscle proteome at the muscle level, the tissue of economic importance in meat production. Furthermore, the identification of proteins that change their abundance in response to SWL using proteomics can contribute to increased stock productivity of relevant interest to animal production. We identified 668 proteins of the sheep proteome. We demonstrate that the following proteins are affected by restricted nutritional conditions: ferritin heavy chain; immunoglobulin V lambda chain; transgelin; fatty acid synthase; glutathione S-transferase A2; dihydrodiol dehydrogenase 3-like. Furthermore, S100-A10, Serpin A3-5-like and Catalase are proteins that changed their abundance in response to SWL. Nevertheless, it is important to highlight that Catalase values for the merino breed were close to significance and therefore catalase validation is of utmost importance.

The hepatic and skeletal muscle ovine metabolomes as affected by weight loss: a study in three sheep breeds using NMR-metabolomics.

Sheep are a valuable resource for meat and wool production. During the dry summer, pastures are scarce and animals face Seasonal Weight Loss (SWL), which decreases production yields. The study of breeds tolerant to SWL is important to understand the physiological mechanisms of tolerance to nutritional scarcity, and define breeding strategies. Merino, Damara and Dorper sheep breeds have been described as having different levels of tolerance to SWL. In this work, we assess their liver and muscle metabolomes, and compare the responses to feed restriction. Ram lambs from each breed were divided into growth and feed restricted groups, over 42 days. Tissue metabolomes were assessed by 1H-NMR. The Dorper restricted group showed few changes in both tissues, suggesting higher tolerance to nutritional scarcity. The Merinos exhibited more differences between treatment groups. Major differences were related to fat and protein mobilization, and antioxidant activity. Between the Damara groups, the main differences were observed in amino acid composition in muscle and in energy-related pathways in the liver. Integration of present results and previous data on the same animals support the hypothesis that, Dorper and Damara breeds are more tolerant to SWL conditions and thus, more suitable breeds for harsh environmental conditions.

The Effect of Weight Loss on the Muscle Proteome in the Damara, Dorper and Australian Merino Ovine Breeds.

Seasonal Weight Loss (SWL) is an important constraint, limiting animal production in the Tropics and the Mediterranean. As a result, the study of physiological and biochemical mechanisms by which domestic animal breeds respond to SWL is important to those interested in animal breeding and the improvement thereof. To that end, the study of the proteome has been instrumental in gathering important information on physiological mechanisms, including those underlying SWL. In spite of that, little information is available concerning physiological mechanisms of SWL in production animals. The objective of this study was to determine differential protein expression in the muscle of three different breeds of sheep, the Australian Merino, the Dorper and the Damara, each showing different levels of tolerance to weight loss (low, medium and high, respectively). Per breed, two experimental groups were established, one labeled "Growth" and the other labeled "Restricted." After forty-two days of dietary treatment, all animals were euthanized. Muscle samples were then taken. Total protein was extracted from the muscle, then quantified and two-dimensional gel electrophoresis were conducted using 24 cm pH 3-10 immobiline dry strips and colloidal coomassie staining. Gels were analyzed using Samespots® software and spots of interest were in-gel digested with trypsin. The isolated proteins were identified using MALDI-TOF/TOF. Results indicated relevant differences between breeds; several proteins are suggested as putative biomarkers of tolerance to weight loss: Desmin, Troponin T, Phosphoglucomutase and the Histidine Triad nucleotide-binding protein 1. This information is of relevance to and of possible use in selection programs aiming towards ruminant animal production in regions prone to droughts and weight loss.

Influence of feed restriction on the wool proteome: a combined iTRAQ and fiber structural study.

Seasonal weight loss is the main limitation to animal production worldwide, significantly affecting the productivity of milk, meat and wool farms, particularly in drought-prone areas of the world where most of the large-scale wool production farms are located. Although the effect of nutritional status on wool quality parameters has been extensively studied, little is known on how it affects wool protein composition. Here, a proteomic approach has been applied to study changes in fiber structure and protein composition in wool from merino sheep subjected to experimentally induced weight loss. Results indicate that there is a significant reduction in the fiber diameter of wool from the animals on a restricted diet over a 42-day period. At the same time, significant increases in the expression of the high sulfur protein KAP13.1 and proteins from the high glycine-tyrosine protein KAP6 family in the wools from the animals on the restricted diet were also detected. Such findings have strong implications for the wool industry that favors finer wool. BIOLOGICAL SIGNIFICANCE: Seasonal weight loss caused by poor pasture availability has strong effects on wool productivity parameters and quality traits. In this work we determine that experimentally induced weight loss causes a decrease in fiber diameter associated with an increase in the level of high sulfur protein KAP13.1 and proteins from the high glycine-tyrosine protein KAP6 family. The implication of this is that decreasing the fiber diameter of the wool by this process could result in a fiber reduced prickle but with reduced wearability and appearance retention.

Does the fat tailed Damara ovine breed have a distinct lipid metabolism leading to a high concentration of branched chain fatty acids in tissues?

Fat tailed sheep breeds are known for their adaptation to nutritional stress, among other harsh production conditions. Damara sheep, native to Southern Africa, have recently been exported to other areas of the world, particularly Australia, aiming to produce lamb in semi-arid regions. Damaras have a unique hanging fat tail, a fat depot able to be mobilized under nutritional stress. In this article we perform an in-depth characterization of the fatty acid profiles of the fat tail in underfed and control Damara rams. Profiles were very similar between experimental groups, with the exception of palmitic acid (16:0) that was lower (P = 0.014) in underfed animals. However, the most striking result was the very high proportions of non-terminal branched chain fatty acids found in the fat tail adipose tissue, as well as the gastrocnemius muscle of Damara rams. The muscle of Dorper and Merino rams used in the same experiment did not present non-terminal branched chain fatty acids, suggesting that Damara rams have a unique lipid metabolism. Herein, we interpret this trait relating it to a higher ability of Damara sheep to digest fibrous fodder and to putative differences in the propionate metabolism by comparison to other sheep breeds.