2. Universidad Cardenal Herrera-CEU

Nutritional ecology seeks to unravel the extensive web of nutritional links that directs animals in their interactions with their ecological and social environments. European rabbit (Oryctolagus cuniculus) populations its endemic locations are declining and it is considered a keystone species of the Mediterranean ecosystem prompteing the interest in its conservation. The main aim of this study was to determine the nutritional composition of the diet of European rabbits through the relative and absolute chemical composition of the gastric content. To address this objective, gastric content was collected from 80 European rabbits in a Mediterranean area for the analysis of its chemical composition. To this end, gastric content was analyzed for dry matter (DM), organic matter (OM), ash, crude protein (CP), highly digestible nonnitrogenous nutrients (HDNN), neutral detergent fibre (NDF), acid detergent fibre (ADF) and lignin. The rabbits were divided into two groups: EMPTY and FULL, depending on the level of stomach filling, directly related to food intake. Our results revealed a positive correlation between the rabbits weight with DM in the gastric content, total gastric content with DM in the gastric content, and DM in gastric content with all chemical parameters analysed. The mean relative values obtained were 8.8%, 25.5%, 40.4% and 25.4%, for ash, CP, NDF and HDNN, respectively. Moreover, EMPTY rabbits had both a proportional (+19%, p = 0.002 and –40%; p = 0.004, on NDF and HDNN, respectively) and absolute (−38%, p = 0.014, –52%; p = 0.012, −52%; p = 0.011 and +83%; p = 0.008 for OM, ash, HDNN, and lignin, respectively) different proportion of nutrients in gastric contents than FULL animals. Since there is a connection between this availability and the fitness of this species, understanding the chemical composition of the rabbit's diet can be utilised to delve into its biology. Our study provides information that will help elucidate the factors affecting the chemical composition of the gastric content of European rabbits to assist land use planners and conservationists in identifying sites for conservation in Mediterranean ecosystems.

The ecological interest in the European rabbit (Oryctolagus cuniculus) has grown since it was declared an endangered species. Nutrition is fundamental in its dispersion and the key to its success. This is the main reason why knowledge of their nutritional preferences/requirements could play a fundamental role in rabbit biology and, therefore, in their conservation. The objectives of the work will be to elucidate how age and sex affect both nutritional preferences/requirements, and the nutrient optimisation of European rabbits. To address this gap, 70 wild European rabbits were studied. The rabbits were divided according to their age: adult (n = 43) and young (n = 27). Two groups were differentiated according to the adult rabbits' sex: females (n = 28) and males (n = 15). We analysed the relative (%) and absolute (g) chemical composition of the rabbits' gastric contents: dry matter (DM), ashes (ASH), crude protein (CP), ether extract (EE), neutral detergent fibre (NDF), acid detergent fibre (ADF), and lignin, hemicelluloses and celluloses, highly digestible nonnitrogenous nutrients (HDNN: fat, starch, and soluble fibre), well from the blood was analysed with plasmatic urea nitrogen (PUN), nonesterified fatty acid (NEFA) and glucose (GLU). As a nutrient optimisation measure, the following indices between blood metabolites and stomach content were calculated: PUN/CP total content, NEFA/DM total content and GLU/DM total content ratios. Our works showed that age and sex affected the nutritional strategy of rabbits. Regarding age: adults showed lower CP (−14%; p = 0.0217) and higher HDNN (+21%; p = 0.0399) relative content than young rabbits, and absolute amount of most nutrients: DM (+59%; p = 0.001), OM (+43%; p = 0.0049), ASH (+54%; p = 0.0085), Hemicelluloses (+73%; p = 0.0084), Cellulose (+27%; p = 0.0452), and HDNN (+63%; p = 0.0012). In addition, adults showed better nutrient optimisation. Sex did not affect the relative chemical composition of the gastric content, but it showed a clear higher gastric content by females (+85%; p < 0.0001) and higher intake of most of the absolute chemical components: DM (+64%; p < 0.001), CP (+56%; p = 0.0005), OM (+58%; p = 0.0001), ASH (+44%; p = 0.0123), HDNN (+39%; p = 0.001), NDF (+59%; p = 0.001), ADF (+64%; p = 0.0003), lignin (+82%; p = 0.0036) and cellulose (+58%; p = 0.0002). Finally, we observed that females had better nutrient optimisation than males. This works supports the idea that feeding ecology and nutrition are particularly relevant to reproductive success and fitness in wild animals.

Ecological nutrition aims to unravel the extensive web of nutritional links that drives animals in their interactions with their ecological environments. Nutrition plays a key role in the success of European wild rabbit (Oryctolagus cuniculus) and could be affected by the breeding status of the animals and reflected in the metabolome of this species. As nutritional needs are considerably increased during pregnancy and lactation, the main objective of this work was to determine how the breeding status (pregnant and lactating) of European wild rabbit does affects nutritional requirements and their metabolome (using targeted and untargeted metabolomics), aiming to find a useful biomarker of breeding status and for monitoring nutritional requirements. To address this gap, 60 wild European rabbits were studied. Animals were divided according to their breeding status and only pregnant (n = 18) and lactating (n = 11) rabbit does were used (n = 29 in total). The body weight and length of each animal were analyzed. The relative and absolute chemical composition of the gastric content and whole blood sample were taken, and targeted and untargeted metabolomics were analyzed. As a main result, there were no differences in biometric measurements, gastric content, and targeted metabolomics, except for live weight and nonesterified fatty acids (NEFA), as pregnant animals showed higher live weight (+12%; p = 0.0234) and lower NEFA acid levels (−46%; p = 0.0262) than lactating females. Regarding untargeted metabolomics, a good differentiation of the metabolome of the two breeding groups was confirmed, and it was proven that pregnant animals showed higher plasmatic levels of succinic anhydride (3.48 more times; p = 0.0236), succinic acid (succinate) (3.1 more times; p = 0.0068) and propionic acid (3.98 more times; p = 0.0121) than lactating animals. However, lactating animals showed higher levels of N-[(3a,5b,7b)-7-hydroxy-24-oxo-3-(sulfoxide) cholan-24-yl]-Glycine (cholestadien) (2.4 more times; p < 0.0420), 4-maleyl-acetoacetate (MAA) (3.2 more times; p < 0.0364) and irilone (2.2 more times; p = 0.0451) than pregnant animals, any of these metabolites could be used as a potential biomarker. From these results, it can be concluded that the most notable changes were observed in the metabolome of individuals, with most of the changes observed being due to energy and protein mobilisation.