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.

European wild rabbit (Oryctolagus cuniculus) has been defined as a keystone species in the Mediterranean ecosystem. Rabbits have been classed as “endangered” by the IUCN within their native range. In this sense, animal nutrition may play a fundamental and limiting role in the conservation of wild species. The overarching goal of ecological nutrition is to unravel the extensive web of nutritional links that direct animals in their interactions with their ecological environments. The main aim of this work was to evaluate the effect of different feed intake, geographic location, animal sex, and reproductive stage on glucose, non-esterified fatty acids (NEFA), and plasmatic urea nitrogen (PUN), albumin, glutamate, and total protein metabolites. Additionally, we examined the potential of these metabolites as biomarkers. Full stomach contents and blood samples were collected from European wild rabbits (n = 89) for the analysis of the metabolites described above. Our work shows that the levels of these metabolites are affected by the sex of the animals, as well as by their reproductive stage (glucose, NEFA and albumin). There were signs of better optimisation of resources by females than by other groups of animals. These data may be interesting in the study of nutritional components that could be affecting physiological state of this species.

European wild rabbit (Oryctolagus cuniculus) populations have drastically reduced, and recently, rabbits have been classed as “endangered” by the IUCN. This animal plays an important ecological role in Mediterranean ecosystems and its introduction could significantly contribute to ecological restoration. Rabbits have high nutrient requirements that apparently cannot be covered in all ecosystems, and there are clues that nutrition can limit their abundance and density. On the other hand, some studies reflect the effects of food availability on the metabolomic status of other animal species, but there are no specific studies on this keystone species. The main aim of this work is to find biomarkers to assess the previous levels of ingestion of European rabbits (Oryctolagus cuniculus). To address this gap, gastric content and blood samples were collected from European rabbits (n = 99) in a Mediterranean area for the analysis of glucose, non-esterified fatty acids (NEFA), plasmatic urea nitrogen (PUN), albumin, glutamate and total protein metabolites. Depending on their previous feed intake (gastric content and the ratio between the gastric content and the weight of the animal), the animals were divided into two groups (lower and normal previous feed intake). Our work shows that the metabolomic profiles of the animals were affected. Levels of glucose (+82%; p = 0.0003), NEFA (􀀀61%; p = 0.0040) and PUN (+139%; p < 0.001) were different in the animals with lower previous feed intake than the animals with normal previous feed intake. This work summarises that metabolic phenotype can be interesting when seeking to discover the limiting nutrients and food availability in diets that could affect the ecological fitness and conservation of European wild rabbits. It is important to mention that in this work, only the effects on six different metabolites have been analysed and more studies are necessary to complement the knowledge of possible metabolites that indicate the level of ingestion in this species and others. These (and new) biomarkers could be used as a tool to provide information about individual or population characteristics that other physiological parameters cannot detect, improving the conservation physiology field.