The study of changes within the key agents regulating metabolism during genetic upgrading because of selection can contribute to an improved understanding of genomic and physiological relationships. This may lead to increased efficiencies in animal production. These changes, regarding energy and protein metabolic saving mechanisms, can be highlighted during food restriction periods. In this study, a 20% weight reduction was induced in two rabbit breeds: New Zealand white, a selected meat producer (Oryctolagus cuniculus cuniculus), and Iberian wild rabbit (Oryctolagus cuniculus algirus), with the aim of determining differential protein expression in the gastrocnemius muscle within control (ad libitum) and restricted diet experimental animal groups, using techniques of two-dimensional gel electrophoresis and peptide mass fingerprinting. Results show that L-lactate dehydrogenase, adenylate kinase, beta enolase and alpha enolase, fructose bisphosphate aldolase A and glyceraldehyde 3-phosphate dehydrogenase, which are enzymes involved in energy metabolism, are differentially expressed in restricted diet experimental animal groups. These enzymes are available to be further tested as relevant biomarkers of weight loss and putative objects of manipulation as a selection tool towards increasing tolerance to weight loss. Similar reasoning could be applied to 2D gel electrophoresis spots corresponding to the important structural proteins tropomyosin beta chain and troponin I. Finally, a spot identified as mitochondrial import stimulation factor seems of special interest as a marker of undernutrition, and it may be the object of further studies aiming to better understand its physiological role.

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The effect of feed restriction on gene expression of regulatory enzymes of intermediary metabolism was studied in two sheep breeds (Australian Merino and Dorper) subjected to two nutritional treatments: feed restriction (85% of daily maintenance requirements) and control (ad libitum feeding), during 42 days. The experimental animals (ram lambs) were divided into four groups, n 5 5 (Australian Merino control (MC), Australian Merino Restriction (MR), Dorper control (DC) and Dorper Restriction (DR)). After the trial, animals were sacrificed and samples were taken from liver tissue to quantify glucose levels and gene expression of relevant intermediary metabolism enzymes (phosphofructokinase (PFK), pyruvate kinase (PK), phosphoenolpyruvate carboxykinase, fructose 1,6-bisphosphatase, glucose-6-phosphatase, glycogen synthase (GS), fatty acid synthase (FAS), glutamate dehydrogenase (GDH) and carbamoyl phosphate synthase (CPS)) through real-time PCR. During the experimental period, the MR animals lost 12.6% in BW compared with 5.3% lost by the Dorper lambs. MC and DC rams gained, respectively, 8.8% and 14% during the same period. Within the Dorper breed, restricted feed animals revealed a significant decrease over controls in the transcription of PFK (1.95-fold) and PK (2.26-fold), both glycolytic enzymes. The gluconeogenesis showed no change in the feed restricted animals of both breeds. DR feed group presented a significant decrease over the homologous Merino sheep group on GS. In both experimental breeds, FAS mRNA expression was decreased in restricted feed groups. GDH expression was decreased only in the DR animals (1.84-fold) indicating a reduced catabolism of amino acids in these animals. Finally, CPS was significantly (P , 0.05) higher in the Dorper sheep, indicating a facilitated urea synthesis in this breed. These results indicate a better adaptation of metabolic intermediate regulatory enzymes and hepatic glucose production of Dorper sheep to feed restriction concurring with the BW results in the experimental groups. ImplicationsThe relevant negative impact in livestock production due to seasonal dry periods within tropical and arid climate regions highlights the need to understand genetic and metabolic adaptation mechanisms to periodical feed restriction. Our results concerning changes in BW and genomic expressions of intermediary metabolism regulatory enzymes of ram lambs of Dorper and Australian Merino breeds subjected to ad libitum and to 85% maintenance energy requirements show that Dorper have a better adaptation to feed restriction than the Merino sheep. Our findings can contribute to future research guidelines leading to improved genetic selection toward livestock adaptation to harsh environments.

This work aims at the identification of relevant intermediate metabolism enzymes contributing to improved meat production due to genetic selection. A wild rabbit (WR) breed and a highly meat selected breed (New Zealand (NZ) rabbit) were used. Food restriction was used as an experimental condition so as to enhance differences within the metabolic pathways under study. During a period of 30 days, NZ and WR experimental breeds were subjected to, respectively, 40% and 60% ad libitum food restriction leading to 17.7% and 21.1% initial weight. Hepatic glycolytic, lipidic and protein regulatory enzyme activity, transcriptional and metabolite levels were determined. Insulin-like growth factor (IGF-1), triiodothyronine, and cortisol were also evaluated. In the glycolytic pathways, the NZ control rabbits presented a higher phosphofructokinase and pyruvate kinase activity level when compared to the WR, while the latter group showed a higher expression of glycogen synthase, although with less glycogen content. In the nitrogen metabolism, our results showed a lower activity level of glutamate dehydrogenase in WR when subjected to food restriction. Within the lipid metabolism, results showed that although WR had a significantly higher mRNA hepatic lipase, non-esterified fatty acid levels were similar between the experimental groups. NZ rabbits presented a better glycemia control and greater energy substrate availability leading to enhanced productivities in which triiodothyronine and IGF-1 played a relevant role.Keywords: enzyme activity, intermediate metabolism, rabbits, regulatory enzymes, feed restriction ImplicationsThe effects of feed restriction on the intermediate metabolism of breeds of rabbits selected for meat production induce an increase over non-selected rabbits on the efficiency of metabolic systems responsible for the homeostasis with reflection on the glycemia maintenance levels and on the excretion process of protein catabolism products. According to our results, seasonal draft periods inducing food restriction might not impair the use of highly productive breeds in upgrading animal production. Further studies are necessary regarding these metabolic pathways, including integrated genomic, transcriptomic and metabolomic approaches leading to results with practical consequences that are specifically required to upgrade livestock production in areas submitted to seasonal feedstuff shortage.

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.

With this work we show other qualities (higher levels of the omega-3 fatty acids, EPA, DPA and DHA) of Damara meat that might present this breed as an interesting alternative for animal production in semi-arid climates.

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