Active monitoring of long-eared owl (Asio otus) nestlings reveals widespread exposure to anticoagulant rodenticides across different agricultural landscapes.

The widespread use of anticoagulant rodenticides (ARs) poses a worldwide threat to farmland wildlife. These compounds accumulate in tissues of both target and non-target species, potentially endangering both direct consumers and their predators. However, investigations on ARs in blood of free-ranging predatory birds are rare. Here, the long-eared owl (Asio otus) has been used as a model predator to assess AR exposure in different agricultural landscapes from a Mediterranean semiarid region. A total of 69 owlets from 38 nests were blood-sampled over 2021 and 2022, aiming to detect AR residues and explore factors that determine their exposure, such as land uses. In addition, prothrombin time (PT) test was conducted to assess potential effects of AR contamination. Overall, nearly all the samples (98.6 %) tested positive for at least one compound and multiple ARs were found in most of the individuals (82.6 %). Among the ARs detected, flocoumafen was the most common compound (88.4 % of the samples). AR total concentration (ΣARs) in blood ranged from 0.06 to 34.18 ng mL-1, detecting the highest levels in the most intensively cultivated area. The analysis of owl pellets from 19 breeding territories showed relevant among-site differences in the contribution of rodents and birds into the diet of long-eared owls, supporting its high dietary plasticity and indicating AR presence at multiple trophic levels. Moreover, a positive and significant correlation was found between ΣARs and PT (Rho = 0.547, p < 0.001), which demonstrates the direct effect of ARs on free-living nestlings. Our results provide a preliminary overview of AR exposure in a little-studied owl species inhabiting agricultural and rural landscapes. Despite the low detected levels, these findings indicate widespread exposure -often to multiple compounds- from early life stages, which raises concern and draws attention to an ongoing and unresolved contamination issue.

Predation impact on threatened spur-thighed tortoises by golden eagles when main prey is scarce.

A reduction in adult survival in long-living species may compromise population growth rates. The spur-thighed tortoise (Testudo graeca) is a long-lived reptile that is threatened by habitat loss and fragmentation. Golden eagles (Aquila chrysaetos), whose breeding habitats overlap that of tortoises, may predate them by dropping them onto rocks and breaking their carapaces. In SE Spain, the number of golden eagles has increased in the last decades and the abundance of their main prey (i.e., rabbits Oryctolagus cuniculus) has decreased. Our aims were to 1) describe the role of tortoises in golden eagles' diet, and 2) estimate the predation impact of golden eagles on tortoises in eagles' territories and in the regional tortoise population. We collected regurgitated pellets and prey remains under eagle nests and roosts, and obtained information on tortoise abundance and population structure and rabbit abundance. We found that tortoises were an alternative prey to rabbits, so that eagles shifted to the former where the latter were scarce. The average predation rate on tortoises was very low at the two studied scales. However, eagles showed a marked selection for adult female tortoises, which led the tortoise sex ratio to be biased towards males in those eagle territories with higher tortoise predation. Whether this may compromise the spur-thighed tortoise long-term population viability locally deserves further attention.

Blood Toxic Elements and Effects on Plasma Vitamins and Carotenoids in Two Wild Bird Species: Turdus merula and Columba livia.

Birds have historically suffered adverse effects by toxic elements, such as As, Pb, Hg, and Cd. However, reports on exposure to a wide range of elements, including rare earth elements and other minor elements of emerging concern, and the potential consequences for wildlife are still scarce. This study evaluates blood concentrations of 50 elements and their related effects on lutein and vitamin levels in the Eurasian blackbird (Turdus merula) and wild rock pigeon (Columba livia), inhabiting different scenarios of contaminant exposure. Blood concentrations of As, Cd, and Pb (and Mn in T. merula) were increased in both species captured in the mining area, compared to the control site. T. merula also showed increased As, Cd, and Pb concentrations in blood in the agricultural-urban area, as compared to the control area, together with the highest Hg levels, which could be related to agricultural practices and industrial activities. Decreases of 33 and 38% in the plasma retinol levels in T. merula inhabiting the mining and the agricultural-urban areas, respectively, as compared to the control site, were associated with increased Pb, As, and Cd exposure. This could be due to a metal-driven suppressive effect in retinol metabolism and/or its over-use for coping with metal-related oxidative stress.

Blood concentrations of 50 elements in Eagle owl (Bubo bubo) at different contamination scenarios and related effects on plasma vitamin levels.

Some metals and metalloids (e.g. Pb, Hg, Cd and As) are well-known for their bioaccumulation capacity and their toxic effects on birds, but concerns on other minor elements and rare earth elements (ME and REE) are growing due to their intensive use in modern technology and potential toxicity. Vitamins and carotenoids play essential roles in nestling growth and proper development, and are known to be affected by the metals classically considered as toxic. However, we are unaware of any attempts to evaluate the exposure to 50 elements and related effects in plasma vitamins and carotenoids in raptor species. The main goals of this study are: (i) to assess the exposure to 50 elements (i.e. classic toxic elements, trace elements, REE and ME) in nestling Eagle owls (Bubo bubo) inhabiting three differently polluted environments (mining, industrial and control areas) in southeastern Spain, and (ii) to evaluate how element exposure affects plasma vitamin and carotenoid levels, hematocrit and body measurements (mass and wing length) of the individuals. Our results show that local contamination in the mining area contributes to increased blood concentrations of Pb, As and Tl in nestlings, while diet differences between control and mining/industrial areas may account for the different levels of Mn, Zn, and Sr in blood, and lutein in plasma. Plasma tocopherol levels were increased in the mining-impacted environment, which may be a mechanism of protection to prevent toxic element-related oxidative stress. Plasma α-tocopherol was enhanced by 20% at blood Pb concentrations ≥8 ng/ml, and nestlings exhibited up to 56% increase in α-tocopherol levels when blood Pb concentrations reached 170 ng/ml. Tocopherol seems to be a sensitive biomarker under an exposure to certain toxic elements (e.g. Pb, As, Tl).

Toxic elements in blood of red-necked nightjars (Caprimulgus ruficollis) inhabiting differently polluted environments.

Toxic metals have been widely reported in avian tissues due to their well-known accumulation capacity and adverse effects. However, rare earth elements (REE) and other minor elements (ME) are becoming a new threat due to their use in modern technology. Presently, exposure data are limited and no studies have been reported in wildlife. The order Caprimulgiformes is among the most understudied groups of birds lacking blood ecotoxicological data. One major constraint is the small blood sample volume that can be collected to ensure animal welfare, which limits analyses. In order to shed light on these collective issues, we sampled 48 red-necked nightjars (Caprimulgus ruficollis) inhabiting three different scenarios of contaminant exposure (agricultural-urban area, n = 15; mining area, n = 17; and control area, n = 16) in southeastern Spain, and report for the first time concentrations of 50 elements (i.e. trace elements, ATSDR's list toxic elements, REE and ME) using a recently developed technique able to analyze them by ICP-MS in very small volumes of blood (130 µL). Concentrations of As, Cd, Pb and Mn were significantly higher in individuals captured at the mining area compared to the other sites. Lead levels in the mine site were of particular concern since it was in the range of blood concentrations related to subclinical/clinical effects in other species, and in our study were associated with decreased hematocrit values (up to 44% hematocrit depression at blood concentrations >1000 ng/ml w.w.). Moreover, additive effects related to metal cocktail exposure in the mining area could be expected. Age and gender-related differences in blood concentrations were found for some elements. Even though most REE and ME concentrations were close to the LOQ, some of these emerging contaminants may trigger sublethal effects that, together with the ATSDR's list toxic elements, need to be carefully evaluated in a future study.

Spatiotemporal variations of organochlorine pesticides in an apex predator: Influence of government regulations and farming practices.

BACKGROUND: Intensification of agricultural practices has caused several negative effects to the environment. The use of fertilizers and pesticides may alter geochemical cycles or cause direct wildlife intoxication. Detrimental effects of organochlorine pesticides (OCPs) have forced the authorities to ban or restrict its use. This study evaluates the variation in levels of OCPs in a sentinel species in relation to changes in government regulations and the spatial configuration of agricultural practices around the nests. METHODS: Between 2003 and 2007, we analysed OCP levels in 256 blood samples of Eurasian Eagle-owl (Bubo bubo) chicks nesting in area of intensive commercial agriculture with historical frequent use of pesticides, in South-eastern Spain. We studied year-to-year variations in OCP concentrations and their relation with land use configuration around raptor nests by Generalized Linear Mixed Models (GLMM). RESULTS: OCPs were detected in 100% samples surveyed in 2003 and 2004, while dropped to 27% in 2005, 6.8% in 2006 and 6.3% in 2007, coinciding with the ban of OCPs. The presence of the main OCPs was related to agricultural practices. In particular, endosulfan and lindane were related to irrigated crops and urban areas, while DDT-related compounds and dieldrin were associated with dry land farming. CONCLUSIONS: OCP concentrations in blood samples of Eurasian Eagle-owls may respond quickly to the implementations of new regulations about the use of agricultural products. This raptor was confirmed as a good sentinel species allowing rapid detection of changes in pesticides use.

Use of classical bird census transects as spatial replicates for hierarchical modeling of an avian community.

New monitoring programs are often designed with some form of temporal replication to deal with imperfect detection by means of occupancy models. However, classical bird census data from earlier times often lack temporal replication, precluding detection-corrected inferences about occupancy. Historical data have a key role in many ecological studies intended to document range shifts, and so need to be made comparable with present-day data by accounting for detection probability. We analyze a classical bird census conducted in the region of Murcia (SE Spain) in 1991 and 1992 and propose a solution to estimating detection probability for such historical data when used in a community occupancy model: the spatial replication of subplots nested within larger plots allows estimation of detection probability. In our study, the basic sample units were 1-km transects, which were considered spatial replicates in two aggregation schemes. We fit two Bayesian multispecies occupancy models, one for each aggregation scheme, and evaluated the linear and quadratic effect of forest cover and temperature, and a linear effect of precipitation on species occupancy probabilities. Using spatial rather than temporal replicates allowed us to obtain individual species occupancy probabilities and species richness accounting for imperfect detection. Species-specific occupancy and community size decreased with increasing annual mean temperature. Both aggregation schemes yielded estimates of occupancy and detectability that were highly correlated for each species, so in the design of future surveys ecological reasons and cost-effective sampling designs should be considered to select the most suitable aggregation scheme. In conclusion, the use of spatial replication may often allow historical survey data to be applied formally hierarchical occupancy models and be compared with modern-day data of the species community to analyze global change process.

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.

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.