Antonio J. García-Fernández

An overview of existing raptor contaminant monitoring activities in Europe

Biomonitoring using raptors as sentinels can provide early warning of the potential impacts of contaminants on humans and the environment and also a means of tracking the success of associated mitigation measures. Examples include detection of heavy metal-induced immune system impairment, PCB-induced altered reproductive impacts, and toxicity associated with lead in shot game. Authorisation of such releases and implementation of mitigation is now increasingly delivered through EU-wide directives but there is little established pan-European monitoring to quantify outcomes. We investigated the potential for EU-wide coordinated contaminant monitoring using raptors as sentinels. We did this using a questionnaire to ascertain the current scale of national activity across 44 European countries. According to this survey, there have been 52 different contaminant monitoring schemes with raptors over the last 50years. There were active schemes in 15 (predominantly western European) countries and 23 schemes have been running for >20years; most monitoring was conducted for >5years. Legacy persistent organic compounds (specifically organochlorine insecticides and PCBs), and metals/metalloids were monitored in most of the 15 countries. Fungicides, flame retardants and anticoagulant rodenticides were also relatively frequently monitored (each in at least 6 countries). Common buzzard (Buteo buteo), common kestrel (Falco tinnunculus), golden eagle (Aquila chrysaetos), white-tailed sea eagle (Haliaeetus albicilla), peregrine falcon (Falco peregrinus), tawny owl (Strix aluco) and barn owl (Tyto alba) were most commonly monitored (each in 6-10 countries). Feathers and eggs were most widely analysed although many schemes also analysed body tissues. Our study reveals an existing capability across multiple European countries for contaminant monitoring using raptors. However, coordination between existing schemes and expansion of monitoring into Eastern Europe is needed. This would enable assessment of the appropriateness of the EU-regulation of substances that are hazardous to humans and the environment, the effectiveness of EU level mitigation policies, and identify pan-European spatial and temporal trends in current and emerging contaminants of concern.

Effects of heavy metals on biomarkers for oxidative stress in Griffon vulture (Gyps fulvus).

Metals are involved in the formation of reactive oxygen species (ROS) which may result in metal-related oxidative stress that can lead to oxidative damage to lipids, DNA and proteins. It is necessary to understand the mechanisms of metal toxicity in wild birds, and the concentrations that cause effects on oxidative stress biomarkers. The aim of this study is to assess the concentrations of lead (Pb), cadmium (Cd), mercury (Hg), copper (Cu) and zinc (Zn) with regards to oxidative stress in blood samples of 66 Griffon vultures (Gyps fulvus) from two areas of the Autonomous Community of Valencia (East of Spain). The two study areas (Alcoy n=36 and Cinctorres n=30) were selected as random locations of interest that had not yet been studied, and are feeding stations where supplementary food, mainly of pork origin, is provided for vultures. Given that the two study areas are not considered polluted sites, we expected to find low metal concentrations. However, there are no known threshold concentrations at which metals can affect antioxidant systems, and low metal levels may have an effect on antioxidant biomolecules. In this study, since sampling was done at the beginning of the hunting season, the low Pb levels found in most Griffon vultures from Alcoy and Cinctorres (median=12.37 and 16.26μg/dl, respectively) are suggestive of background levels usually found in vultures that feed on pork carcasses all year round. The ingestion of game meat with bullet fragments in carcasses or with Pb shots embedded in the flesh could be the cause of the high blood Pb concentrations found in three vultures from Cinctorres (83, 290 and 362μg/dl). Griffon vultures feeding in Cinctorres had enhanced CAT and GST activities and tGSH concentrations, which may be interpreted as protective response against the higher TBARS levels. This study provides threshold concentrations at which metals affect antioxidant system derived from 66 samples of Griffon vulture. Blood Cd concentrations greater than 0.05μg/dl produced an induction of 33% in GPx and of 44% in CAT activity in erythrocytes of vultures from Alcoy. Hg concentrations in blood higher than 3μg/dl produced an induction of 10% in SOD activity. Concentrations of Pb above 15µg/dl in blood produced an inhibition of 12.5% in GPx and 11.3% in CAT activity, and a TBARS induction of 10.7% in erythrocytes of Griffon vultures.

Heavy metals in tissues from loggerhead turtles (Caretta caretta) from the southwestern Mediterranean (Spain)

Cadmium, lead, copper, and zinc were measured in tissues of 21 loggerhead turtles (Caretta caretta) from the southwestern Mediterranean coastline. Mean concentrations (dry weight) of essential elements (Zn and Cu) were 107 and 21.6 microg/g in liver, 27.9 and 3.8 microg/g in kidney, 65.4 and 5.0 microg/g in pectoral muscle, 11.1 and 3.45 microg/g in brain, and finally 19.2 microg/g and undetected in bone, respectively. Mean concentrations of heavy metals (Cd and Pb) were 23.4 and 2.8 microg/g in liver, 31.5 and 0.5 microg/g in kidney, 0.2 and 0.3 microg/g in pectoral muscle, 0.2 and 0.7 microg/g in brain, and undetected and 1.2 microg/g in bone, respectively. Metal concentrations were similar to other studies conducted on Mediterranean turtles. However, cadmium concentrations varied widely among individuals, which has been associated with potential sources of cadmium in Mediterranean Sea. This is the first study into metal accumulation in tissues of loggerhead turtle from Spanish Mediterranean coastline.

Feathers as a Biomonitoring Tool of Polyhalogenated Compounds: A Review.

Feathers have many advantages that make them an excellent nondestructive tool for monitoring polyhalogenated compounds (PHCs). This paper proposes a review on the PHCs in feathers and factors influencing the pollutant load. Special attention has given to external contamination and the main analytical methods used to detect these compounds in feathers. Some authors have found strong and significant correlations between the concentrations of PHCs in feathers and internal tissues, providing positive expectations for their future use in the field of ecotoxicology. However, changes in diet, time elapsed between the previous molt period and sampling, sample size, and/or external contamination have been suggested as possible causes to explain the lack of correlations reported in some studies. Further studies with newly grown feathers and blood samples would be required in order to clarify this issue. Although atmospheric deposition has been reported as cause of external contamination, preening oil seems to be the most relevant factor contributing to this process. Unfortunately, washing techniques tested to date are not able to effectively remove the surface contamination from barbs and shafts, and therefore, it is necessary to develop methods able to discriminate between internal and external contamination. Finally, in this review, deposition rate is proposed as a measurement unit, as this allows comparisons between different parts of the same feather, as well as between different feathers.

Raptor Ecotoxicology in Spain: A Review on Persistent Environmental Contaminants

Abstract Initial studies on the pressure from environmental contaminants on raptor populations in Spain date back to the 1980s, and they have been carried out from a range of viewpoints using a range of sentinel raptor species. However, there is no national monitoring scheme, and therefore the research carried out has been sporadic both spatially and temporally. The exposure to metals has not varied over time, except in the case of lead, whose concentration in eggs and tissues has diminished. In general, the concentrations of metals detected in raptor samples from Spain are generally low and not sufficient to produce toxic effects. Excepting DDT and DDE, most organochlorine-based pesticides in raptors from Spain have diminished over the last 2 decades. The concentrations of DDE found in the eggs of various species could in part explain problems in the reproductive success of raptors in Spain.

Alterations of the glutathione-redox balance induced by metals in CHO-K1 cells.

The effects of cadmium (Cd(2+)), mercury (Hg(2+)), lead (Pb(2+)), copper (Cu(2+)) and nickel (Ni(2+)) on the glutathione (GSH)-redox cycle were assessed in CHO-K1 by the neutral red uptake inhibition (NR) assay (NR(6.25), NR(12.5) and NR(25)). Mercury proved to be the most and lead the least toxic of the metals tested. The effects on GSH content and intracellular specific activities of enzymes involved in the GSH-redox balance were measured after a 24-h exposure. Total GSH content increased significantly in cultures exposed to the lowest metal concentration assayed (NR(6.25)), but fell to below control values when exposed to concentrations equivalent to NR(25). Oxidised glutathione content dropped significantly at NR(6.25), while somewhat higher values were obtained for cultures exposed to higher doses. Glutathione peroxidase (Gpx) activities were 1.2-, 1.5-, 1.6-, 2.0- and 2.5-fold higher than untreated controls for cadmium, copper, mercury, nickel and lead, respectively, at concentrations equivalent to NR(6.25). Gpx activity declined at metal concentrations equivalent to NR(12.5) and NR(25). Glutathione reductase activity remained almost unchanged except at low doses of mercury, nickel and lead. Glutathione-S-transferase activity decreased at rising metal concentrations. The results suggest that a homeostatic defence mechanism was activated when cells were exposed to doses equivalent to NR(6.25) while the ability of the cells to respond weakened as the dose increased. A close relationship was also observed between metal cytotoxicity, total GSH content and the dissociation energy of the sulphur-metal bonds. These facts confirm the involvement of antioxidant defence mechanisms in the toxic action of these ions.

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.

A review on exposure and effects of arsenic in passerine birds

UNLABELLED Arsenic (As) is a metalloid of high concern because of its toxic effects for plants and animals. However, it is hard to find information on this metalloid in passerines. This review presents a comprehensive overview of As exposure and effects in birds, and more particularly in passerines, as a result of an extensive search of the literature available. Internal tissues are the most frequently analyzed matrices for As determination in passerines (37.5% of the reviewed studies used internal tissues), followed by feathers and eggs (32.5% each), feces (27.5%), and finally blood (15%). A clear tendency is found in recent years to the use of non-destructive samples. Most studies on As concentrations in passerines have been done in great tit (Parus major; 50%), followed by pied flycatcher (Ficedula hypoleuca; 22.5%). Some factors such as diet and migratory status are crucial on the interspecific differences in As exposure. More studies are needed to elucidate if intraspecific factors like age or gender affect As concentrations in different tissues. The literature review shows that studies on As concentrations in passerines have been done mainly in the United States (30%), followed by Belgium (22.5%), and Finland (20%), making evident the scarce or even lack of information in some countries, so we recommend further research in order to overcome the data gap, particularly in the southern hemisphere. Studies on humans, laboratory animals and birds have found a wide range of effects on different organ systems when they are exposed to different forms of As. This review shows that few field studies on As exposure and effects in passerines have been done, and all of them are correlative so far. Arsenic manipulation experiments on passerines are recommended to explore the adverse effects of As in free-living populations at similar levels to those occurring in the environment. CAPSULE This review summarizes the most interesting published studies on As exposure and effects in passerines.

Organochlorine residues in booted eagle (Hieraaetus pennatus) and goshawk (Accipiter gentilis) eggs from southeastern Spain

Most organochlorine (OC) use has been banned in Spain, but these compounds are persistent and may still adversely affect predatory birds. Data generally are lacking, however. Residues of hexachlorobenzene, alpha-hexachlorocyclohexane, lindane, hexachloro-octahydro-epoxy-dimethanonaphthalene, DDT, dichlorodiphenyldichloroethane, dichlorodiphenyldichloroethylene (DDE), and polychlorinated biphenyls (PCBs) were measured in 22 failed eggs of booted eagles (Hieraaetus pennatus) and goshawks (Accipiter gentilis) from southeastern Spain; both species are declining in this region. Hexachloro-octahydro-epoxy-dimethanonaphthalene, DDE, and sum PCB congener concentrations were significantly higher in booted eagle than in goshawk eggs, and an inverse relationship was found between shell thickness and DDE concentrations in booted eagles. Organochlorides may have been associated with the failure of some booted eagle eggs, but concentrations in booted eagle and goshawk eggs decreased over the period during which populations have dwindled. Thus, although OCs may be a contributory factor, they are unlikely to be the primary cause of the recent population declines in southeastern Spain.

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.