Targeted and untargeted metabolomic profiles in wild rabbit does (Oryctolagus cuniculus) of different breeding states (pregnant and lactating).

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.

Dietary type (carnivore, herbivore and omnivore) and animal species modulate the nutritional metabolome of terrestrial species.

Ecometabolomics could be implemented as a powerful tool in molecular ecology studies, but it is necessary to know the baseline of certain metabolites and understand how different traits could affect the metabolome of the animals. Therefore, the main objective of this study was to provide values for the nutritional metabolome profile of different diet groups and animal species, as well as to study the differences in the metabolomic profile due to the effect of diet type and species. To achieve this goal, blood samples were taken from healthy animals (n = 43) of different species: lion (Panthera leo), jaguar (Panthera onca), chimpanzee (Pan troglodytes), bison (Bison bison), gazelle (Gazella cuvieri) and fallow deer (Dama dama), and with different types of diet (carnivore, herbivore and omnivore). Each blood sample was analysed to determine nutritional metabolites. The main results this study provides are the nutritional metabolic profile of these animals based on the type of diet and the animal species. A significant effect of the dietary type was found on nutritional metabolite levels, with those metabolites related to protein metabolism (total protein and creatine) being higher in carnivores. There is also an effect of the species on nutritional metabolites, observing a metabolome differentiation between lion and jaguar. In the case of herbivores, bison showed higher levels of uric acid and cholesterol, and lower urea levels than gazelle and fallow deer. More molecular ecology studies are needed to further the knowledge of the metabolism of these animals.

Prevalence and factors associated with Leishmania spp. and Toxoplasma gondii infections in apparently healthy horses in Eastern Spain.

Leishmaniasis and toxoplasmosis are two of the most common parasitic zoonoses. Leishmaniasis is endemic to 98 countries around the world, whereas toxoplasmosis is widely distributed throughout the world, causing significant health expenditure. Horses can play a relevant role in the transmission of the disease, being a silent reservoir, as clinical signs are not common. Serum samples from 166 horses living in eastern Spain (Mediterranean basin) were analysed to determine the presence of antibodies against Leishmania spp. and T. gondii by ELISA (Enzyme-linked Immunosorbent Assay.) The risk factors evaluated were the geographical area and the relative humidity and average temperature, and epidemiological factors such as sex, reproductive status, age, breed, morphotype, living with other domestic animals, use and access to the outdoors. Seroprevalence of Leishmania spp. and T. gondii infection was found 28.92%, and 16.27% respectively, whereas co-infection of the two parasites was found only in two males. Leishmania seroprevalence was high in castrated males and several mesodolichomorphic equine breeds used for teaching, as well as in outdoor animals. The most elevated seroprevalence was found in winter with higher levels of rainfall, whereas high seroprevalence of T. gondii was found in crossbreeding animals and those used for breeding. High seroprevalence of Leishmania spp. and T. gondii was found in horses of the Mediterranean basin. These data suggest that horses can act as a silent reservoir and that this species has high potential for transmission to humans, outdoor animals and in geographical areas with high average rainfall.

A Trial Comparing Growing Rabbits Differing in 18 Generations of Selection for Growth Rate Reveals a Potential Lack of Effectiveness in the Genetic Selection Progress.

A total of 338 weaned rabbits (from the R line, selected for post-weaning growth rate) were used to evaluate the response to 18 generations of selection for increased growth rate on rabbit performance. Animals were obtained from two vitrified populations of the R line: R19V, belonging to the 18th generation (n = 165), and R37V, belonging to the 36th generation (n = 173), were allocated in individual and collective pens (178 and 160, respectively). A fattening trial was conducted from weaning (28 d of age until 63 d of age). During the trial, the body weight (BW), daily feed intake (DFI), average daily gain (ADG) and feed conversion ratio (FCR) were weekly monitored. Additionally, mortality and morbidity were daily registered. On days 49 to 53, an apparent faecal digestibility trial was also performed (12 animals per generation). Our results indicate that the generation of selection for growth rate did not affect mortality and morbidity. There were no differences in the diet digestibility according to the generation of selection. Regarding performance traits, R37V animals showed higher global BW (+6.7%; p = 0.0011) than R19V animals. R37V animals showed the same BW at weaning; however, R37V animals showed higher BW values in the last three weeks compared with R19V animals. Animals from the R37V generation also showed a higher DFI from 56 to 63 d of age (+12%; p = 0.0152) than R19V animals. However, there were no differences in global ADG and FCR between generations. These results indicate that the selection for growth rate in growing rabbits has slowed down, suggesting a lack of effectiveness in the genetic progress.

How Does Nutrition Affect the Epigenetic Changes in Dairy Cows?

Dairy cows require a balanced diet that provides enough nutrients to support milk production, growth, and reproduction. Inadequate nutrition can lead to metabolic disorders, impaired fertility, and reduced milk yield. Recent studies have shown that nutrition can affect epigenetic modifications in dairy cows, which can impact gene expression and affect the cows' health and productivity. One of the most important epigenetic modifications in dairy cows is DNA methylation, which involves the addition of a methyl group to the DNA molecule. Studies have shown that the methylation status of certain genes in dairy cows can be influenced by dietary factors such as the level of methionine, lysine, choline, and folate in the diet. Other important epigenetic modifications in dairy cows are histone modification and microRNAs as regulators of gene expression. Overall, these findings suggest that nutrition can have a significant impact on the epigenetic regulation of gene expression in dairy cows. By optimizing the diet of dairy cows, it may be possible to improve their health and productivity by promoting beneficial epigenetic modifications. This paper reviews the main nutrients that can cause epigenetic changes in dairy cattle by analyzing the effect of diet on milk production and its composition.

Prevalence and Factors Related to Leishmania infantum Infection in Healthy Horses (Equus caballus) from Eastern Spain.

Leishmaniosis is a zoonosis caused by Leishmania spp., an intracellular protozoan parasite. This parasite is transmitted by sandflies, and the disease is endemic in the Mediterranean basin. In recent years, the number of species which could be a reservoir of the parasite is increased. One of the most relevant species is the horse, due to their contact with humans and ability to control the disease, thus being a possible silent reservoir. In this study, we have analyzed the prevalence and factors related to L. infantum infection in healthy horses in the Mediterranean region. Epidemiological data and serum samples were obtained from 167 apparently healthy horses, and the presence of L. infantum was evaluated via the ELISA method and real-time PCR. The results show 27.5% of prevalence and that the main factors related to infection are equine breed, morphotype, outdoor living, use, and season. In conclusion, the prevalence of L. infantum infection in apparently healthy horses from eastern Spain (Mediterranean basin) is elevated. To control this zoonosis, it would be advisable to carry out more studies on this and other species that could be silent reservoirs of the parasite, as well as carry out measures such as the use of repellents on a regular basis.

Nutritional ecology of European rabbit (Oryctolagus cuniculus): Factors affecting chemical composition of gastric content.

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.

Environmental gaseous pollutants are related to increase of acute coronary syndrome in Valencia region, Spain / Los contaminantes gaseosos ambientales están relacionados con el aumento del síndrome coronario agudo

Introduction and aims Environmental pollution are one of the most relevant risk factors to atherosclerosis. To know awareness about the importance of urban air pollution as a trigger for hospital admission due to acute coronary syndrome (ACS), this study analyzed levels of different gaseous air pollutants in the air and its correlation with number of ACS. Methods Epidemiological data of patients admitted for ACS in five towns during the years 2006–2008 were recovered. Clinical data regarding admissions for ACS were obtained from the hospital admission services. Measures of seven air contaminants were recovered from the environmental stations. Mixed model including sex, age, location, and the average levels of air pollutants contaminants as fixed effects and its interaction were performed. Results The incidence rate of ACS is higher in man than woman, and higher in older people than young. The maximum ACS were in the last trimester of the year, was the most elevated levels of gaseous pollutants have been found. Levels of NO2, NO, and CO are positively correlated between them, and negatively correlated with O3 levels. All air pollutants analyzed increase the number of ACS hospital admission in the five locations evaluated. Conclusions Levels of gaseous pollutants are related between them, being the levels of NO2, NO, and CO, positively correlated, and negatively correlated with levels of O3. Number of ACS hospital admission increases with levels of five air gaseous pollutants studied (AU)

Introducción y objetivos La contaminación ambiental es uno de los factores de riesgo más relevantes para la aterosclerosis. Para conocer la importancia de la contaminación del aire urbano como desencadenante del ingreso hospitalario por síndrome coronario agudo (SCA), este estudio analizó los niveles de diferentes contaminantes atmosféricos gaseosos en el aire y su correlación con el número de SCA. Métodos Se recuperaron datos epidemiológicos de pacientes ingresados por SCA en 5 municipios durante los años 2006 a 2008. Los datos clínicos relativos a los ingresos por SCA se obtuvieron de los servicios de ingreso hospitalario. Se recuperaron medidas de 7 contaminantes del aire de las estaciones ambientales. Se realizó un modelo mixto que incluye sexo, edad, ubicación y los niveles promedio de contaminantes atmosféricos como efectos fijos y su interacción. Resultados La tasa de incidencia de SCA es mayor en hombres que en mujeres, y mayor en personas mayores que en jóvenes. La mayoría de los SCA se dieron en el último trimestre del año, cuando se encontraron niveles más elevados de gases contaminantes. Los niveles de NO2, NO y CO se correlacionan positivamente entre ellos y negativamente con los niveles de O3. Todos los contaminantes atmosféricos analizados aumentan el número de ingresos hospitalarios por SCA en las 5 localidades evaluadas. Conclusiones Los niveles de contaminantes gaseosos se relacionan entre sí, estando los niveles de NO2, NO y CO correlacionados positivamente entre sí y negativamente con los niveles de O3. El número de ingresos hospitalarios por SCA aumenta con los niveles de 5 contaminantes gaseosos del aire estudiados (AU)

Effect of Increasing the Methionine Level and Reducing the Threonine Level in the Diet of Fast-Growing Rabbits.

The main aim of this work was to evaluate a novel combination of the first limiting amino acids (lysine, methionine, and threonine) in fast-growing rabbits (combination MHL, shown to minimise levels of plasmatic urea nitrogen in previous research, medium level of lysine, high level of methionine, and low level of threonine) and compare it with current recommendations (combination MMM, medium level of all amino acids). A total of 165 weaned rabbits (28 d old) from a paternal line selected for growth rate were used in the growth trial. The effect of the diet on apparent fecal digestibility, as well as the apparent and true ileal digestibility, were studied. Nutrient retention was also determined. Although no differences in digestibility between diets were observed, animals fed with Diet MHL improved the global average daily gain (+2.3 g/d; p = 0.0482) and feed conversion ratio (-0.10; p = 0.0229). Animals fed with Diet MMM reduced the protein-to-energy ratio retained (p = 0.0086). In conclusion, Diet MHL promoted an improvement in growth traits in a paternal line. Consequently, we propose its levels of 6.4, 5.4, and 5.0 g/kg DM of true ileal digestible lysine, methionine, and threonine, respectively.

The nutritional strategy of European rabbits is affected by age and sex: Females eat more and have better nutrient optimisation.

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.

Immunological profile of two canine breeds in an endemic region of Leishmania infantum.

Visceral leishmaniosis is the one of the most important protozoal zoonoses in Europe, and it is caused by Leishmania infantum, an intracellular protozoan parasite. The disease is endemic in dogs in the Mediterranean area. The main goal of this work is to correlate the levels of several cytokines linked to immune response against L. infantum infection in two canine breeds. Thirty-one Boxer and twenty-eight Ibizan Hound dogs living in the Valencian Community (East coast of Spain) were analyzed for the presence of anti-Leishmania antibodies in serum by IFAT test. Cytokines IFN-γ, TNF-α, IL-2, IL-6, IL-8, and IL-18 were determined by ELISA commercial tests. The levels of IFN-γ, IL-2, and IL-18 in our study, cytokines linked to a cellular immune response, were higher (p < 0.05) in the Ibizan Hound breed; IL-6 levels were higher, although not significant, and only levels of IL-8 were higher in Boxer than in Ibizan Hound. No expression of TNF-α was found. These results corroborate that Ibizan Hound can develop a protective response against canine leishmaniosis, while Boxer is a susceptible breed. The study of immunological aspects in the different canine breeds may represent a useful tool in the prediction of the disease.

Genetic haplotypes associated with immune response to Leishmania infantum infection in dogs.

Leishmaniasis is a zoonotic parasitic disease, and the main reservoir of the parasite is the dog, although recent years have seen an increase in other mammalian species. In the Mediterranean region, where it is an endemic disease, it is caused by the species Leishmania infantum. The Ibizan hound, an autochthonous breed of this region, appears to have a genetic resistance to parasitic infection, whereas other canine breeds, such as the Boxer, are susceptible to infection. These differences are related to the differentiated activation of the immune response, with the Ibizan hound activating the Th1 immune response, whereas the Boxer breed triggers the Th2 immune response. Cytokine levels and genomic haplotypes of several genes involved in the immune response were analysed in twenty-eight Ibizan hound (resistant canine breed model) and twenty-four Boxer (susceptible canine breed) without clinical signs in the Mediterranean region. Cytokine levels were analysed by ELISA commercial kits and haplotypes were studied using CanineHD DNA Analysis BeadChip including 165,480 mapped positions. The results show 126 haplotypes associated with differential immune response in dogs. Specifically, haplotypes in IL12RB1, IL6R, CIITA, THEMIS, NOXA1, HEY2, RAB38, SLC35D2, SLC28A3, RASEF and DAPK1 genes are associated with serum levels of IFN-γ, IL-2, IL-8, and IL-18. These results suggest that the resistance or susceptibility to Leishmania infantum infection could be a consequence of haplotypes in several genes related to immune response. Future studies are needed to elucidate the relationship of these haplotypes with immune response and gene expression regulation.

Environmental gaseous pollutants are related to increase of acute coronary syndrome in Valencia region, Spain.

INTRODUCTION AND AIMS: Environmental pollution are one of the most relevant risk factors to atherosclerosis. To know awareness about the importance of urban air pollution as a trigger for hospital admission due to acute coronary syndrome (ACS), this study analyzed levels of different gaseous air pollutants in the air and its correlation with number of ACS. METHODS: Epidemiological data of patients admitted for ACS in five towns during the years 2006-2008 were recovered. Clinical data regarding admissions for ACS were obtained from the hospital admission services. Measures of seven air contaminants were recovered from the environmental stations. Mixed model including sex, age, location, and the average levels of air pollutants contaminants as fixed effects and its interaction were performed. RESULTS: The incidence rate of ACS is higher in man than woman, and higher in older people than young. The maximum ACS were in the last trimester of the year, was the most elevated levels of gaseous pollutants have been found. Levels of NO2, NO, and CO are positively correlated between them, and negatively correlated with O3 levels. All air pollutants analyzed increase the number of ACS hospital admission in the five locations evaluated. CONCLUSIONS: Levels of gaseous pollutants are related between them, being the levels of NO2, NO, and CO, positively correlated, and negatively correlated with levels of O3. Number of ACS hospital admission increases with levels of five air gaseous pollutants studied.

Effect of selection for growth rate on the rabbit (Oryctolagus cuniculus) immune system and its response after experimental Staphylococcus aureus infection.

The aim of the work was to evaluate if genetic selection for daily gain may affect the immune system. Two experiments were performed. The first one involved 80 rabbit females and their first two litters to explore the effect of selection on the ability of animals to maintain immune competence. Two generations from a line selected for average daily gain (ADG) were evaluated (VR19 generation 19th, n = 43; VR37 generation 37th, n = 37). In females, the effect of selection and its interaction with physiological state were not significant for any trait. In litters, the selection criterion increased the granulocyte to lymphocyte ratio. The second experiment involved 73 19-week-old females (VR19, n = 39; VR37, n = 34) to explore the effect of genetic selection on immune response after S. aureus infection. The VR37 rabbit females had lower counts for total lymphocytes, CD5+, CD4+, CD8+, CD25+, monocytes, the CD4+/CD8+ ratio and platelets than those of VR19 (-14, -21, -25, -15, -33, -18, -11 and -11%, respectively; P < 0.05). VR37 had less erythema (-8.4 percentage points; P < 0.05), fewer nodules (-6.5 percentage points; P < 0.05) and a smaller nodule size (-0.65 cm3 on 7 day post-inoculation; P < 0.05) compared to VR19. Our study suggests that genetic selection for average daily gain does not negatively affect the maintenance of a competent immune system or the ability to establish immune response. It seems that such selection may improve the response to S. aureus infections.

Epigenetic Alterations in Canine Malignant Lymphoma: Future and Clinical Outcomes.

Canine malignant lymphoma is a common neoplasia in dogs, and some studies have used dogs as a research model for molecular mechanisms of lymphomas in humans. In two species, chemotherapy is the treatment of choice, but the resistance to conventional anticancer drugs is frequent. The knowledge of molecular mechanisms of development and progression of neoplasia has expanded in recent years, and the underlying epigenetic mechanisms are increasingly well known. These studies open up new ways of discovering therapeutic biomarkers. Histone deacetylases and demethylase inhibitors could be a future treatment for canine lymphoma, and the use of microRNAs as diagnosis and prognosis biomarkers is getting closer. This review summarises the epigenetic mechanisms underlying canine lymphoma and their possible application as treatment and biomarkers, both prognostic and diagnostic.

Interleukin 6 and interferon gamma haplotypes are related to cytokine serum levels in dogs in an endemic Leishmania infantum region.

BACKGROUND: The Ibizan Hound is a canine breed native to the Mediterranean region, where leishmaniasis is an endemic zoonosis. Several studies indicate a low prevalence of this disease in Ibizan Hound dogs, whereas other canine breeds present a high prevalence. However, the underlying molecular mechanisms still remain unknown. The aim of this work is to analyse the relationship between serum levels of cytokines and the genomic profiles in two canine breeds, Ibizan Hound (resistant canine breed model) and Boxer (susceptible canine breed model). METHODS: In this study, we analyse the haplotypes of genes encoding cytokines related to immune response of Leishmania infantum infection in twenty-four Boxers and twenty-eight Ibizan Hounds apparently healthy using CanineHD DNA Analysis BeadChip including 165,480 mapped positions. The haplo.glm extension of haplo.score was used to perform a General Linear Model (GLM) regression to estimate the magnitude of individual haplotype effects within each cytokine. RESULTS: Mean levels of interferon gamma (IFN-γ), interleukin 2 (IL-2) and IL-18 in Boxer dogs were 0.19 ± 0.05 ng/ml, 46.70 ± 4.54 ng/ml, and 36.37 ± 30.59 pg/ml, whereas Ibizan Hound dogs present 0.49 ± 0.05 ng/ml, 64.55 ± 4.54 ng/ml, and 492.10 ± 31.18 pg/ml, respectively. The GLM regression shows fifteen haplotypes with statistically significant effect on the cytokine serum levels (P < 0.05). The more relevant are IL6-CGAAG and IFNG-GCA haplotypes, which increase and decrease the IL-2, IL-8 and IFN-γ serum levels, respectively. CONCLUSIONS: Haplotypes in the IFNG and IL6 genes have been correlated to serum levels of IFN-γ, IL-2 and IL-18, and a moderate effect has been found on IL8 haplotype correlated to IL-8 and IL-18 serum levels. The results indicate that the resistance to L. infantum infection could be a consequence of certain haplotypes with a high frequency in the Ibizan Hound dog breed, while susceptibility to the disease would be related to other specific haplotypes, with high frequency in Boxer. Future studies are needed to elucidate whether these differences and haplotypes are related to different phenotypes in immune response and expression gene regulation to L. infantum infections in dogs and their possible application in new treatments and vaccines.

Immunological and genomic characterization of Ibizan Hound dogs in an endemic Leishmania infantum region.

BACKGROUND: The Ibizan Hound is a canine breed native to the Mediterranean region, where leishmaniosis is an endemic zoonosis. Several studies indicate low prevalence of this disease in these dogs but the underlying molecular mechanism remains unknown. METHODS: In this study, qualitative immunological and genomic profiles of this breed have been analyzed. RESULTS: Our analysis shows relevant differences between the cytokine serum profile of Ibizan Hound dogs and previously published data from other canine strains. Additionally, several genetic risk variants related to the immune response, regulation of the immune system, and genes encoding cytokines and their receptors have been studied. The most relevant genes that presented such fixed polymorphisms were IFNG and IL6R. Other variants with frequencies ≥ 0.7 were found in the genes ARHGAP18, DAPK1, GNAI2, MITF, IL12RB1, LTBP1, SCL28A3, SCL35D2, PTPN22, CIITA, THEMIS, and CD180. Epigenetic regulatory genes such as HEY2 and L3MBTL3 showed also intronic polymorphisms. CONCLUSIONS: Our analysis and results indicate that the regulation of immune responses is different in Ibizan Hounds compared to other breeds. Future studies are needed to elucidate whether these differences are related to the low prevalence of L. infantum infection in the Ibizan Hound.

Unravelling the Role of Metabolites for Detecting Physiological State of Wild Animals: European Rabbit's (Oryctolagus cuniculus) Case.

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.

Serum levels and genetic variations of cytokines in two canine breeds (Ibizan hound and boxer) in the Mediterranean region, in terms of Leishmania infantum infection.

Leishmaniosis, endemic to the Mediterranean area and considered to be the most important zoonosis in Europe, is caused by Leishmania infantum, an intracellular protozoan parasite transmitted by sandflies Phlebotomus. On the one hand, the Ibizan hound, a canine breed native of the Mediterranean region has been described to present a great resistance to the leishmaniosis parasitic infection. On the other hand, other breeds, including boxer, present high prevalence, and severity of the infection. In this study, serum levels of cytokines related to immune response together with the genetic variants found in genes encoding these cytokines have been interrogated in dogs of these two canine breeds (Ibizan hound and boxer) living in the Mediterranean region. A total of 59 apparently healthy dogs were sampled with their corresponding epidemiological data. Serum levels of cytokines were interrogated by ELISA commercial kits and the genetic analysis was carried out with the commercial CanineHD BeadChip and subsequent bioinformatic analysis. The results showed higher levels of IFN-γ, IL-2, and IL-18 and lower levels of IL-8 in Ibizan hound dogs compared to boxer ones. Genomic analysis revealed several genetic variants fixed in the Ibizan hound samples, including IFNG, IL2, IL6R, and IL12RB1 genes, which could explain these differences.

Immune and Genomic Analysis of Boxer Dog Breed and Its Relationship with Leishmania infantum Infection.

Leishmaniosis, one of the most important zoonoses in Europe, is caused by Leishmania infantum, an intracellular protozoan parasite. This disease is endemic in the Mediterranean area, where the main reservoir is the dog. Several studies indicate a possible susceptibility to L. infantum infection with clinical signs in some canine breeds. One of them is the boxer breed, which shows a high prevalence of disease. In this study, immunological and genomic characterization of serum samples from boxer dogs living in the Mediterranean area were evaluated to analyze the immune response and the possible genetic explanation for this susceptibility. Serum levels of cytokines IFN-γ, IL-2, IL-6, IL-8, and IL-18 were determined by ELISA commercial tests, while the genotyping study was performed using the CanineHD DNA Analysis BeadChip. The results show relevant differences in the serum levels of cytokines compared to published data on other canine breeds, as well as sequence changes that could explain the high susceptibility of the boxer breed to the disease. Concretely, polymorphic variants in the CIITA, HSF2BP, LTBP1, MITF, NOXA1, PKIB, RAB38, RASEF, TLE1, and TLR4 genes were found, which could explain the susceptibility of this breed to L. infantum infection.