Non-invasive assessment by feathers of lead exposure and its relationship with stress hormones in bearded vultures from the Alps
R. Mateo, IREC-CSIC- UCLM / Grupo de Toxicología de Fauna Silvestre; L. Giménez-Lozano, IREC Instituto de Investigación en Recursos Cinegéticos; L. Monclus, UAB; I. Champly, ASTERS; M. Lopez-Bejar, UAB
A reintroduction project of bearded vulture or lammergeier (Gypaetus barbatus) has been carried out in the Alps since 1987. There are several factors that can affect the success of this reintroduction, and one of them is exposure to lead from the ammunition used for hunting. An exposure to lead, even at sub-lethal levels, can be a stressful situation that implies an increase in circulating corticosterone levels. In birds that are rare and difficult to capture, blood sampling for both lead and corticosterone analysis can be a difficult task and with some risk for birds. For this reason, the development of non-invasive methodologies to study exposure to pollutants and their respective biomarkers is being directed towards the analysis of easy-to-collect samples, such as feathers moulted by birds. The objectives of this study are to quantify the degree of exposure to lead that the bearded vultures have in the Alps by analysing moulted feathers found in the field, relating them to corticosterone levels in the same feather and defining the factors that affect the fluctuations of these two components along transverse segments of feathers. The analysis of the segments of 20 different feathers from different bearded vultures revealed that the abnormal exposure to lead (probably due to the ingestion of ammunition) had a prevalence of 15% (three feathers with levels above 2 μg/g of Pb in rachis), and that the annual incidence of such exposure was 30% (a feather with an abnormal exposure during its development of approximately two months). The concentration of lead in the segments of rachis feathers was negatively related to the concentration of corticosterone, and was significantly influenced by the collection area and the individual (feather). In the present study we observed that exposure to elevated levels of lead can occur in a high percentage of individuals in a population throughout the year, which in the case of the lammergeier of the Alps can entail a risk to the sustainability of the population if this exposure reach lethal levels.
MO079
Post mortem stability of phase I and II biotransformation enzymes in the liver of kelp gull Larus dominicanus
J.J. Mattos, Universidade Federal de Santa Catarina / NEPAQ, Departamento de Aquicultura; D.d. Lima, Universidade Federal de Santa Catarina / Bioquimica; B. Righetti, Universidade Federal de Santa Catarina / NEPAQ-CCA; L.O. Villas Boas, V.H. Dias, Universidade Federal de Santa Catarina / CCB; S. Costa-Silva, C. Kolesnikovas, M. Antonelli, Associação R3 Animal; K. Luchmann, Santa Catarina State University / Engenharia de Pesca; A. Bainy, Universidade Federal de Santa Catarina / Bioquimica
The measurement of biomarker responses to chemical contaminants in wild organisms represents a powerful tool in environmental monitoring programs. However, getting biological samples suitable for biomarker analysis may be challenging, since some programs rely on samples collected from carcasses. Shorter periods from death to analysis of biochemical biomarkers provides more accurate results, but how these parameters change at longer post mortem intervals remains unclear. This study evaluated the post mortem stabilityof key biotransformation enzymes (glutathione S-transferase, GST and 7-etoxyresorufin-O-deetilase, EROD) in the liver of kelp gull Larus dominicanus for periods up to 24 hours. Liver tissue of two euthanized animals was sectioned into several 1-cm3 cubes and stored in individual closed tubes at 25 oC for 0, 1, 2, 3, 6, 12, 18 and 24 h post mortem before liquid nitrogen freezing. Cytosolic and microsomal fractions were obtained from 150 mg of each sample individually and used for GST and EROD
measurements, respectively. GST activity proved to be stable after 24 h (85-90% of initial activity). EROD activity decreased abruptly after the first hour post mortem for both animals. After 3 hours EROD activity presented 65 to 71 % of initial activity and 28 to 50% of its initial activity after 6h, showing an exponential decrease along post mortem period. The estimated half-life for EROD varied from 2.8 to 5.2 hours post mortem. Our results indicate that time elapsed since death until sample collection plays an essential role for biotransformation enzymes, especially concerning EROD activity. GST seemed to be more resistant to degradation over time, and it thus appears possible to make valid GST activity measurement in selected post mortem liver tissue of kelp gull. Overall, our findings demonstrate that caution is warranted in monitoring programs when comparing biological samples with different intervals between collection and analysis procedures.
MO080
Investigating thyroid disrupting effects of organohalogenated contaminants in White-tailed eagle nestlings
M.E. Løseth, The Norwegian University of Science and Technology / Biology; G.S. Eggen, N. Briels, Norwegian University of Science and Technology / Biology; T. Nygård, T.V. Johnsen, J.O. Bustnes, Norwegian Institute for Nature Research NINA; D. Herzke, NILU - Norwegian Institute for Air Research / FRAM Centre Tromso; G. Poma, G. Malarvannan, University of Antwerp Toxicological Center; A. Covaci, University of Antwerp, Toxicological Center / Toxicological Centre Dep of Pharmaceutical Sciences; B.M. Jenssen, Norwegian University of Science and Technology / Biology; V. Jaspers, Norwegian University of Science & Technology / Biology
White-tailed eagles (WTE; Haliaeetus albicilla) can accumulate a wide range of
organohalogenated contaminants (OHCs), due to their apex trophic position. Their diet consists mainly of fish and seabirds, thus long food chains and a high potential for biomagnification of OHCs. The nestlings can therefore be exposed to high levels of certain OHCs through maternal transfer to the eggs, and later through diet. Concentrations of per- and polyfluoroalkyl substances (PFASs) have recently been shown to exceed those of other legacy OHCs in this species and accordingly required closer attention. Several of these POPs have shown to interfere with endocrine systems in birds, and especially the thyroid system. The thyroid system is important for birds’ thermoregulation, metabolism, growth and development. Assuring appropriate concentrations and actions of the two major thyroid hormones thyroxine (T4) and triiodothyronine (T3) in the circulatory system is therefore especially important in nestlings. The aim of the present study was, for the first time, to investigate the effect of POPs and PFASs accumulation in plasma on thyroid hormones (TH) of nestling white-tailed eagles. We also included the body mass and age to assess influence of biological variables on the TH. Blood plasma samples were obtained from 70 nestlings of white-tailed eagles from two archipelagos in Norway, Smøla (n = 35) and Steigen (n = 35), in the summer of 2015 and 2016. In total, 14 polychlorinated biphenyls (PCBs), 7 organochlorinated pesticides (OCPs), 5 polybrominated diphenyl ethers (PBDEs) and 8 PFASs were quantified in over 50 % of the plasma samples at each location and each year. Our results show higher OHC concentrations in Steigen [median and range; Σ14PCBs: 5.1 ng/ml (1.5 – 59.1 ng/ml), Σ7OCPs: 4.2 ng/ml (1.3 – 52.2 ng/ml), Σ5PBDEs: 0.3 (< 0.1 – 2.6 ng/ml) and Σ8PFASs: 20.8 ng (7.2 – 52.9 ng/ml)], than Smøla [median and range; Σ14PCBs: 3.9 ng/ml (0.8-34.7 ng/ml), Σ7OCPs: 2.4 ng/ml (0.9 – 15.3 ng/ml), Σ5PBDEs:0.2 (< 0.1 – 1.5 ng/ml) and Σ8PFASs: 10.9 ng (4.6 – 46.7 ng/ml)]. The analyses of thyroid hormones have been carried out and the results will be presented at the conference, along with biological parameters and OHCs.
MO081
Assessment of exposure and effects of Hg levels in feathers of White-tailed eagles (Haliaeetus albicilla) and Northern goshawks (Accipiter gentilis) nestlings from Norway
P. Gómez-Ramírez, University of Murcia / Department of Toxicology; J.O. Bustnes, Norwegian Institute for Nature Research / Fram Centre; G.S. Eggen, Norwegian University of Science and Technology / Biology; I. Eulaers, University of Antwerp / Biology; G. Lepoint, University of Liege / Department of Oceanology; J.M. Pérez-García, University of Lleida / Department of Animal Sciences; T.V. Johnsen, Norwegian Institute for Nature Research NINA; A. Garcia-Fernandez, University of Murcia / Sociosanitary Sciences; V. Jaspers, Norwegian University of Science & Technology / Biology
Anthropogenic activities have led to a global increase of Mercury (Hg) in the environment. Due to its toxicity, legislative measures have been taken to reduce the levels. To assess the effectiveness of such restrictions and the current levels of these chemicals in the environment, biomonitoring using birds is very useful. Hg has caused detrimental effects in birds such as haematotoxicity, immunotoxicity and endocrine disruption e.g. suppression of baseline corticosterone. The aim of this study was to assess the exposure to Hg and its effects at the
biochemical/physiological level in White-tailed eagles (WTE) and Northern goshawks (NG) from Norway. Samples were obtained in 2014 from nestling WTE (n=14) and NG (n=11) in northern Norway (Nordland- N 68.30 – 68.47?, E 24.54 – 25.27?- and Troms- N 68.77 – 67.39?, E 20.39 – 23.34?- counties, respectively). Total Hg in feathers, total and free plasma corticosterone levels were analysed, along with following blood clinical chemical parameters (BCCPs): albumin, calcium, phosphorus and γ-glutamyltransferase, lactate dehydrogenase, total proteins, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, cholesterol, triglycerides, creatine kinase, amylase, glucose, creatinine, bilirubin, potassium and uric acid. Stable carbon (13C, 12C) and nitrogen isotopes (15N, 14N) were analysed in body feathers to evaluate inter- and
intraspecific contaminant exposure. Due to the low amount of feather samples, Hg could only be analysed in 13 WTE and 8 NG. Mean ± SD were 0.51 ± 0.34 mg/kg in NG and 3.01 ± 1.34 mg/kg in WTE. The significantly higher levels in WTE than in NG (T (19) = -7.61, p< 0.01) may be related to different dietary input, as confirmed by stable carbon and nitrogen isotope analysis of body feathers. The marine prey of WTE seem to determine the Hg loads, as Hg is known to be abundant in the marine environment. Partial correlations to assess the relations between Hg and
biochemical parameters (corticosterone, BCCPs) showed relations between Hg and aspartate aminotransferase (an enzyme that may increase after liver damage). The effect of mercury on this enzyme seems controversial, as some experimental studies on nestlings of different species have found both positive and negative relations. Moreover, the lack of information on reference values in GH and WTE complicates the interpretation. Acknowledgements: NILS Science and Sustainability.
MO082
Thyroid-related gene expression, hormones, and thyroid gland histology in American kestrels exposed in ovo to two persistent organic pollutants, SCCPs and TBBPA-BDBPE
A. MacLeod, University of Maryland, College Park / Environmental Science and Technology; P.F. Henry, U.S. Geological Survey / Patuxent Wildlife Research Center; K.J. Fernie, Environment & Climate Change Canada / Ecotoxicology and Wildlife Health; N.K. Karouna-Renier, USGS Patuxent Wildlife Research Center /
Patuxent Wildlife Research Center
Highly brominated flame retardants are being replaced by alternative flame retardants such as Tetrabromobisphenol A bis[2,3-dibromopropyl ether] (TBBPA-BDBPE). TBBPA-BDBPE was introduced as a possible substitute for decabromodiphenyl ether (decaBDE), but has shown similar persistency and environmental transport mechanisms. This additive flame retardant is used in plastic products, resins, textiles, paints, and household electronics. Although it is produced only in the US, Israel, and China, TBBPA-BDBPE is detected in environmental samples and wildlife tissues from across the globe. Short-Chain Chlorinated Paraffins (SCCPs) are priority emerging persistent organic pollutants (POPs) identified as chemicals of concern by the Stockholm Convention, Environment and Climate Change Canada, and the U.S. Environmental Protection Agency. SCCPs are used in metal lubricants and coolants in metal cutting, and as plasticizers and flame retardants in plastics and paints. SCCPs are of concern because they bioaccumulate in wildlife and humans, are environmentally persistent, transported globally, and are toxic to aquatic organisms at low concentrations. However, few data are available on the potential adverse effects of
TBBPA-BDBPE and SCCPs in birds. A comparative exposure assessment of these two classes of flame retardants was conducted using egg injections in a non-model species, the American kestrel (Falco sparverius) to assess survival, molecular, biochemical, and endocrine, growth and reproductive endpoints. Analyses conducted on tissue samples from hatchings included thyroid related gene expression in the liver, thyroid hormone levels, and thyroid gland histology. Preliminary results will be presented from the molecular to biochemical to cellular level.
MO083
Bioaccumulation of metals in bats: non-lethal vs lethal sampling to assess risk
J. Alves, R. Mina, A. Alves da Silva, CFE - Centre for Functional Ecology, / Department of Life Sciences, University of Coimbra; T. Natal da Luz, University of Coimbra / Department of Life Sciences, University of Coimbra; J.A. Cabral, P. Barros, CITAB - Centre for Research and Technology of Agro-Environment and Biological Sciences / Laboratory of Applied Ecology, Unversity of Trás-os-Montes e Alto Douro; C.J. Topping, Aarhus University / Department of Bioscience; J. Sousa, University of Coimbra / Department of Life Sciences
More than 22% of bat species are threatened according to the World Conservation Union. Among the reasons responsible for this decline are the environmental changes due to anthropogenic factors, namely habitat loss through agriculture, forestry, urbanization and industrialization, contamination by pesticides and metals, changes in water quality. There is a growing concern about the possible
consequences of environmental contamination in several bat species. Until now, most of the studies on the effects of contaminants in bats are focused on organic contaminants, and the consequences of exposure to other substances (particularly metals), remaining largely unknown. The aim of this study was to evaluate the potential risk of metal contamination in bat species occurring in Portugal, and to evaluate the suitability of non-lethal sampling methods. The concentration of As, Cd, Co, Cr, Cu, Mn, Ni, Pb, Se and Zn was measured in two categories of biological samples (lethal-samples: liver, heart, bone and brain; and non-lethal samples: wing membrane and fur) collected from bat carcasses of four different species (Hypsugo
savii, Nyctalus leisleri, Pipistrellus pipistrellus, Pipistrellus pygmaeus).
Concerning the metal concentration obtained in each sampling tissue, significant differences were found between the concentrations obtained in each species for all the metals (P< 0.05), except for Zinc (P=0.223). Significant differences were also found between organs (P< 0.001), metals (P< 0.001) and the interaction between organs and metals (P< 0.001). Depending on the metal, the organ/tissue that showed the highest concentrations varied, but even so fur and wing presented the highest concentrations of most of the metals. These results support the hypothesis that non-lethal samples may be useful for studies on wildlife ecotoxicology, and may help to define a protocol capable of being applied at large-scale, to investigate the risk of metal accumulation for bats. For this purpose, non-lethal samples are the best option, and as demonstrated by this study can yield reliable results. Our results therefore provide valuable insights for development of further studies, aiming to understand the importance of metals as a cause for some of the observed declines in bat populations worldwide.
MO084
Metallic element composition of egg contents and eggshells of the Kelp Gull Larus dominicanus
J.D. van Aswegen, North West University (Potchefstroom Campus) / Unit for Environmental Science and Management; L. Nel, N. Strydom, Nelson Mandela University; H. Bouwman, North-West University / Unit for Environmental Science and Management
The Swartkops River Estuary near Port Elizabeth, South Africa, is an important recreational, industrial, residential, and ecological asset, but under severe pressure. Seabirds are good indicators of trace elements within their environments. Seabirds tend to feed at different trophic levels at different distances from the land and they are long-lived. Pollutants that have accumulated in the seabirds can be excreted in various ways from the body, one being deposition into eggs. Sixteen eggs of the Kelp Gull (Larus dominicanus) were analysed using ICP-MS (EPA 3050b method) for 30 trace elements, for both the contents and eggshells. We selected five
elements (Cr, Sr, Tl, U, Zn) to compare and assess the absolute and relative compositional patterns in egg contents and shells. Mean concentrations for Cr in eggshells and egg contents were 3.8 and 18 mg/kg dm, for Sr it was 880 and 12 mg/kg dm, for Tl it was 0.00017 and 0.00022 mg/kg dm, for U it was 0.000057 and 0.000084 mg/kg dm, and for Zn it was 2.1 and 62 mg/kg dm, respectively. Of the five elements, only Sr (p = 0.0141) and Tl (p = 0.0013) concentrations showed significant positive regressions between egg contents and eggshells. Chromium and Zn showed a positive association, but the regressions were not significant. Uranium also showed no association. The mean mercury concentration in the contents was 0.38 mg/kg dm, and the maximum was 2.1 mg/kg dm. The Toxic Reference Value for mercury in bird egg contents is 2 mg/kg dm, indicating concern about this element in the Swartkops River Estuary. Additional toxic implications, as well as comparisons with concentrations in other media will be discussed.
MO085
Heavy metals concentrations in Mediterranean Osprey eggs: variations by location, habitat and egg constituent part
F. Monti, University of Siena / Department of Physical Sciences, Earth and Environment; S. Ancora, University of Siena / Physical sciences, Earth and environment; A. Sforzi, Maremma Natural History Museum, Grosseto; N. Bianchi, University of Siena / Department of Physical, Earth and Environmental Sciences; C. Leonzio, University of Siena / Department of Physical Sciences, Earth and Environment
The osprey (Pandion haliaetus) has been historically used world-wide as a sentinel species for the biomonitoring of selected contaminants for aquatic ecosystems. In spite of this, occasional and fragmentary information are available for the species at the Mediterranean scale, where relict and vulnerable populations exist. In this study, we analyzed heavy metals concentrations in osprey eggs from three different populations of the Mediterranean basin (Corsica, Italy and Balearic Islands). In total, 21 unhatched eggs were collected, over a period spanning from 2005 to 2016. Mercury (Hg), Cadmium (Cd) and Lead (Pb) in osprey eggs were analyses with the aim to: (1) evaluate geographical patterns of for possible identification of inputs at the regional scale; 2) to evaluate differences in concentrations between samples from different habitats (marine environments and wetlands); and 3) to investigate any differences in concentrations among different parts of the egg (i.e. content, membrane and shell). Samples from the Balearic Islands showed higher Hg concentrations (1.4 ± 1.2 mg/kg on dry weight basis) compared to other samples. Egg shells from marine environments (Corsica and Balearics) had five times greater [Hg] than those from wetlands. Egg content and membrane showed higher Hg concentration values than those of the shell. On the contrary, for Cd and Pb (though with minor differences) higher concentrations were found in the egg shell. Our study represents a first survey at regional scale and provides a first set of data for the long-term biomonitoring of heavy metals for the vulnerable osprey populations of the Mediterranean basin \n
MO086
Interactive effects of vitamin E and BDE-47 yolk supplementation on morphology and oxidative status of yellow-legged gull embryos
M. Parolini, University of Milan / Department of Environmental Science and Policy; C.D. Possenti, B. De Felice, Università degli Studi di Milano; N. Saino, University of Milano
Oviparous mothers transfer to the eggs components that have both independent and combined effects on offspring phenotype. Functional interactions between egg components, such as antioxidants and contaminants, lead to expect that a change in the concentration of one component has effects on offspring traits that depend on the concentration of other interacting components. However, the combined effects of variation in different egg components are virtually unknown. Bird eggs contain vitamin E (VE), a major antioxidant, and also a variable amount of
maternally-transferred contaminants. Polybrominated diphenyl ethers (PBDEs) are a family of brominated flame retardants that have been widely used as non-reactive additive compounds diverse commercial products. Many monitoring studies have revealed the presence of PBDEs in the biota, which can induce a plethora of adverse effects at different organisms’ life stages, often mediated by the onset of oxidative stress. Although PBDEs have been found in birds and their eggs, the consequences related to the exposure to these chemicals, mainly during early development, are inadequate. In addition, no study has considered that the oxidative stress-related toxicity of these compounds may be counteracted by the presence of antioxidant molecules that mothers allocate to their eggs at the time of laying. The independent consequences of variation in the egg concentrations of VE and PBDEs on offspring phenotype, including morphological and oxidative stress effects, are largely