Mario León-Ortega

Oxidative stress biomarkers in Eurasian eagle owls (Bubo bubo) in three different scenarios of heavy metal exposure.

The main aim of the present study is the assessment of oxidative stress related to metals in the Eurasian eagle owl (Bubo bubo) from three areas (agricultural and rural area, industrial area, and mining area) of Murcia, Southern Spain. Mean blood metal concentrations were Cd=0.07±0.21, Pb=3.27±5.21, Cu=10.62±4.77, Zn=311.47±67.14, Hg=2.32±3.83 μg/dl wet weight. Although individuals from the mining area had significant higher Pb and Hg concentrations, and significant lower glutathione-S-transferase (GST) and catalase (CAT) activities in red blood cells (RBC); the lack of differences in oxidative damage to membrane lipids (TBARS) among areas suggests that the antioxidant capacity of the different populations is able to deal with oxidant species and maintain TBARS levels in the same amount. Despite the low levels of metals, several oxidative stress biomarkers were correlated with metal concentrations. This study provides threshold concentrations at which metals cause effects on the antioxidant system in Eagle owls. Blood Cd concentrations greater than 0.3 μg/dl produced an inhibition in GPx (32%) and CAT (26%) activity in RBC. However, Cd concentrations higher than 0.02 μg/dl were enough to produce an inhibition of these enzymes. Regarding Pb levels, blood concentrations above 2 μg/dl produced an inhibition of 8% and 10.5% in GPx and CAT activities, respectively, in RBC. A depletion of 16% and 4% in tGSH levels was associated with Pb concentrations higher than 15 and 3 μg/dl, respectively, in individuals from the ancient mine site. In addition, Pb concentrations above 2 and 10 μg/dl produced a TBARS induction of 10% and 28%, respectively, in individuals from both the industrial and the mining area. Finally, Hg concentrations greater than 3 and 10 μg/dl resulted in a TBARS induction of 102% and 190%, respectively, in Eurasian eagle owls from the industrial area. Our findings show that Pb may produce effects on oxidative stress biomarkers in Strigiformes at lower concentrations than those typically accepted for considering physiological effects in Falconiformes (20 µg/dl in blood). In addition, we provide new data on Hg and Cd concentrations related to effects in the antioxidant system.

Factors that influence mercury concentrations in nestling Eagle Owls (Bubo bubo)

Mercury (Hg) is a global pollutant that bioaccumulates and biomagnifies in food chains, and is associated with adverse effects in both humans and wildlife. The Hg levels detected in blood obtained from Eagle Owl (Bubo bubo) chicks in Southeast Spain (Murcia) can be considered low (mean Hg concentration in blood from 2006 to 2012 was 36.83 ± 145.58 μg/l wet weigh, n=600), and it is therefore unlikely that Hg pollution can negatively affect their breeding. Positive correlation (r=0.339, p<0.001, n=229) was found to exist between the Hg concentrations in the blood and back feathers of the chicks. We provide a regression equation that could be helpful to estimate blood Hg levels when analyzing Hg concentrations in back feathers. Blood Hg concentrations in Eagle Owls have shown positive correlations with Hg levels in rabbit muscles, more evident in nests from the Northern area (r=0.600, p=0.014, n=16), where rabbits are the main prey of Eagle Owls. The best Linear Mixed Model to explain variations in blood Hg concentrations in nestling Eagle Owls includes year and location within the mining area as variables. The variable year is assigned the largest value of relative importance, followed by the location in the ancient mining sites and then the zone. Rainfalls may have an effect on the temporal differences in the blood Hg concentrations of nestling Eagle Owls. Although the studied region is not considered Hg polluted, the Hg levels were higher in Eagle Owls and European Rabbits from areas within the ancient mining sites as compared to those in the entire region. This result shows that spatial differences in Hg concentrations in Eagle Owls may be affected by local contamination, and that the role of diet composition may be less significant.

Spatial Patterns of Genetic Diversity in Mediterranean Eagle Owl Bubo bubo Populations

Summary. Little information is available on the patterns of genetic connectivity in owls. We studied the genetic structure of the eagle owl Bubo bubo (Linnaeus, 1758) in southeastern Spain at two different spatial scales. Seven microsatellites previously described for this species were used, although only six loci amplified correctly. The observed low genetic variation could be explained by the short dispersal distance, high mortality rate and high degree of monogamy shown by this large nocturnal predator. As expected, the highest genetic isolation was detected in the geographically most isolated population. Significant genetic differentiation was found among study units separated by less than 50 km. The territorial analysis showed interesting connectivity patterns related with the gene flow and turnover rate of the breeding individuals. The lowest genetic diversity was found in the region with the largest population, which could imply incipient inbreeding.

Validation of a QuECheRS method for analysis of neonicotinoids in small volumes of blood and assessment of exposure in Eurasian eagle owl (Bubo bubo) nestlings

Neonicotinoid pesticides have gained great interest in the last years both for agricultural and domestic use. Since the information on their environmental distribution or the effects derived from exposure to ecosystems and biota is scarce, new analytical techniques are being developed for monitoring studies. In this sense, two extraction techniques based on QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) methodology to detect the neonicotinoids authorised in Spain (acetamiprid, clothianidin, dinotefuran, imidacloprid, thiacloprid, nitenpyram and thiamethoxam) were adapted and compared: a) using acetate buffer (AB); and b) using citrate buffer (CB). For detection and quantification, high performance liquid chromatography coupled with time of flight mass spectrometry (HPLC/TOF-MS) was used. The CB method provided a wide range of recoveries (68-134%) and accuracy (4-9%). The AB method provided good recoveries (59-76%, 59% corresponded to clothianidin) precision (4-11%) linearity (0.987-0.998%) and limit of quantification (2-10ng/mL) for all the compounds. To test the effectiveness of the technique, we analysed 30 blood samples of free-ranging nestlings of Eurasian eagle owl (Bubo bubo). The only compound detected, in one nestling from a dry land farming area, was imidacloprid, with a concentration of 3.28ng/mL. To our knowledge, this is the first study of neonicotinoid pesticides in free-ranging birds of prey using non-destructive samples, providing the first insight for biomonitoring studies. Further studies, including toxicokinetics and toxicodynamics, are recommended to assess the risk for these species.

Factors influencing territorial occupancy and reproductive success in a Eurasian Eagle-owl (Bubo bubo) population

Modelling territorial occupancy and reproductive success is a key issue for better understanding the population dynamics of territorial species. This study aimed to investigate these ecological processes in a Eurasian Eagle-owl (Bubo bubo) population in south-eastern Spain during a seven-year period. A multi-season, multi-state modelling approach was followed to estimate the probabilities of occupancy and reproductive success in relation to previous state, time and habitat covariates, and accounting for imperfect detection. The best estimated models showed past breeding success in the territories to be the most important factor determining a high probability of reoccupation and reproductive success in the following year. In addition, alternative occupancy models suggested the positive influence of crops on the probability of territory occupation. By contrast, the best reproductive model revealed strong interannual variations in the rates of breeding success, which may be related to changes in the abundance of the European Rabbit, the main prey of the Eurasian Eagle-owl. Our models also estimated the probabilities of detecting the presence of owls in a given territory and the probability of detecting evidence of successful reproduction. Estimated detection probabilities were high throughout the breeding season, decreasing in time for unsuccessful breeders but increasing for successful breeders. The probability of detecting reproductive success increased with time, being close to one in the last survey. These results suggest that reproduction failure in the early stages of the breeding season is a determinant factor in the probability of detecting occupancy and reproductive success.

Haematocrit and blood biochemical parameters in free-living Eurasian eagle owls (Bubo bubo) from Southeastern Spain: study of age and sex differences

The study of haematocrit and biochemical parameters is considered a useful tool for diagnosis of health status in birds. However, an adequate interpretation requires reference values for each species. Moreover, factors such as age and sex should also be taken into account, as it is well known that they can also influence several parameters. For these reasons, serum biochemical parameters (albumin, calcium, phosphorus, aspartate aminotransferase, alkaline phosphatase, cholesterol, triglycerides, creatine kinase, γ-glutamyltransferase, glucose, lactate dehydrogenase, total proteins and uric acid) were studied in 15 adults (6 males and 9 females) and 258 chicks of free-living Eurasian eagle owl (Bubo bubo), one of the owls with the largest distribution range worldwide. Most blood clinical parameters were higher in nestlings than in adults, but only significantly for alkaline phosphatase, phosphorus and uric acid. On the contrary, γ-glutamyltransferase was significantly lower in nestlings. In adults, sex differences were not significant. Haematocrit was studied in 13 adults (2 males and 11 females) and 162 nestlings, and levels were significantly higher in adults than in nestlings. Because samples were obtained from healthy free-living individuals and the values fall within the range of levels considered physiological for similar species, we suggest that can be used as reference values for Eurasian eagle owls.