Annual abundance of common Kestrels (Falco tinnunculus) is negatively associated with second generation anticoagulant rodenticides.

Rats and mice can damage food and agricultural products as well as transmit diseases, thereby requiring control of their numbers. Application of Second Generation Anticoagulant Rodenticides (SGARs) often reduces rodent numbers locally. However, predators eating rodents, including non-target species, that have consumed SGARs may be secondarily exposed and potentially lethally poisoned. Here we study whether SGARs may have contributed to the widespread population declines of a rodent-eating raptor, the Common Kestrel (Falco tinnunculus) in the UK. We show that 161 (66.8%) of the 241 Kestrels submitted for ecotoxicology tests between 1997 and 2012 had detectable levels of at least one SGAR in their livers. Adult Kestrels had significantly higher prevalence of SGARs than juveniles, suggesting accumulation of SGARs through time. The prevalence and concentrations of individual SGARs in Kestrels were significantly higher in England than in Scotland. SGAR prevalence in Kestrels were positively associated with some land cover types, primarily arable cereals and broad-leaved woodland, and negatively associated with mainly mean elevation, probably reflecting variation in SGAR usage across land cover types. By using volunteer-collected data on national Kestrel abundance 1997-2012, we show that there is a negative correlation between the Kestrel population index in a specific year and the concentration of bromadialone as well as the total SGAR concentration in the same year. Although correlative, this is the first study to provide evidence for a potential population-limiting effect of SGARs on a raptor.

Long-term trends of second generation anticoagulant rodenticides (SGARs) show widespread contamination of a bird-eating predator, the Eurasian Sparrowhawk (Accipiter nisus) in Britain.

Second generation anticoagulant rodenticides (SGARs) are widely used to control rodents around the world. However, contamination by SGARs is detectable in many non-target species, particularly carnivorous mammals or birds-of-prey that hunt or scavenge on poisoned rodents. The SGAR trophic transfer pathway via rodents and their predators/scavengers appears widespread, but little is known of other pathways of SGAR contamination in non-target wildlife. This is despite the detection of SGARs in predators that do not eat rodents, such as specialist bird-eating hawks. We used a Bayesian modelling framework to examine the extent and spatio-temporal trends of SGAR contamination in the livers of 259 Eurasian Sparrowhawks, a specialist bird-eating raptor, in regions of Britain during 1995-2015. SGARs, predominantly difenacoum, were detected in 81% of birds, with highest concentrations in males and adults. SGAR concentrations in birds were lowest in Scotland and higher or increasing in other regions of Britain, which had a greater arable or urban land cover where SGARs may be widely deployed for rodent control. However, there was no overall trend for Britain, and 97% of SGAR residues in Eurasian Sparrowhawks were below 100 ng/g (wet weight), which is a potential threshold for lethal effects. The results have potential implications for the population decline of Eurasian Sparrowhawks in Britain. Fundamentally, the results indicate an extensive and persistent contamination of the avian trophic transfer pathway on a national scale, where bird-eating raptors and, by extension, their prey appear to be widely exposed to SGARs. Consequently, these findings have implications for wildlife contamination worldwide, wherever these common rodenticides are deployed, as widespread exposure of non-target species can apparently occur via multiple trophic transfer pathways involving birds as well as rodents.

Making use of apex predator sample collections: an integrated workflow for quality assured sample processing, analysis and digital sample freezing of archived samples.

Using monitoring data from apex predators for chemicals risk assessment can provide important information on bioaccumulating as well as biomagnifying chemicals in food webs. A survey among European institutions involved in chemical risk assessment on their experiences with apex predator data in chemical risk assessment revealed great interest in using such data. However, the respondents indicated that constraints were related to expected high costs, lack of standardisation and harmonised quality criteria for exposure assessment, data access, and regulatory acceptance/application. During the Life APEX project, we demonstrated that European sample collections (i.e. environmental specimen banks (ESBs), research collection (RCs), natural history museums (NHMs)) archive a large variety of biological samples that can be readily used for chemical analysis once appropriate quality assurance/control (QA/QC) measures have been developed and implemented. We therefore issued a second survey on sampling, processing and archiving procedures in European sample collections to derive key quality QA/QC criteria for chemical analysis. The survey revealed great differences in QA/QC measures between ESBs, NHMs and RCs. Whereas basic information such as sampling location, date and biometric data were mostly available across institutions, protocols to accompany the sampling strategy with respect to chemical analysis were only available for ESBs. For RCs, the applied QA/QC measures vary with the respective research question, whereas NHMs are generally less aware of e.g. chemical cross-contamination issues. Based on the survey we derived key indicators for assessing the quality of biota samples that can be easily implemented in online databases. Furthermore, we provide a QA/QC workflow not only for sampling and processing but also for the chemical analysis of biota samples. We focussed on comprehensive analytical techniques such as non-target screening and provided insights into subsequent storage of high-resolution chromatograms in online databases (i.e. digital sample freezing platform) to ultimately support chemicals risk assessment.

Assessment of contaminants of emerging concern in European apex predators and their prey by LC-QToF MS wide-scope target analysis.

Apex predators are good indicators of environmental pollution since they are relatively long-lived and their high trophic position and spatiotemporal exposure to chemicals provides insights into the persistent, bioaccumulative and toxic (PBT) properties of chemicals. Although monitoring data from apex predators can considerably support chemicals' management, there is a lack of pan-European studies, and longer-term monitoring of chemicals in organisms from higher trophic levels. The present study investigated the occurrence of contaminants of emerging concern (CECs) in 67 freshwater, marine and terrestrial apex predators and in freshwater and marine prey, gathered from four European countries. Generic sample preparation protocols for the extraction of CECs with a broad range of physicochemical properties and the purification of the extracts were used. The analysis was performed utilizing liquid (LC) chromatography coupled to high resolution mass spectrometry (HRMS), while the acquired chromatograms were screened for the presence of more than 2,200 CECs through wide-scope target analysis. In total, 145 CECs were determined in the apex predator and their prey samples belonging in different categories, such as pharmaceuticals, plant protection products, per- and polyfluoroalkyl substances, their metabolites and transformation products. Higher concentration levels were measured in predators compared to prey, suggesting that biomagnification of chemicals through the food chain occurs. The compounds were prioritized for further regulatory risk assessment based on their frequency of detection and their concentration levels. The majority of the prioritized CECs were lipophilic, although the presence of more polar contaminants should not be neglected. This indicates that holistic analytical approaches are required to fully characterize the chemical universe of biota samples. Therefore, the present survey is an attempt to systematically investigate the presence of thousands of chemicals at a European level, aiming to use these data for better chemicals management and contribute to EU Zero Pollution Ambition.

Contrasting long term temporal trends in perfluoroalkyl substances (PFAS) in eggs of the northern gannet (Morus bassanus) from two UK colonies.

We compared long-term (1977 to 2014) trends in concentrations of PFAS in eggs of the marine sentinel species, the Northern gannet (Morus bassanus), from the Irish Sea (Ailsa Craig) and the North Sea (Bass Rock). Concentrations of eight perfluorinated carboxylic acids (PFCAs) and three perfluorinated sulfonates (PFSAs) were determined and we report the first dataset on PFAS in UK seabirds before and after the PFOS ban. There were no significant differences in ∑PFAS or ∑PFSAs between both colonies. The ∑PFSAs dominated the PFAS profile (>80%); PFOS accounted for the majority of the PFSAs (98-99%). In contrast, ∑PFCAs concentrations were slightly but significantly higher in eggs from Ailsa Craig than in those from Bass Rock. The most abundant PFCAs were perfluorotridecanoate (PFTriDA) and perfluoroundecanoate (PFUnA) which, together with PFOA, comprised around 90% of the ∑PFCAs. The ∑PFSAs and ∑PFCAs had very different temporal trends. ∑PFSAs concentrations in eggs from both colonies increased significantly in the earlier part of the study but later declined significantly, demonstrating the effectiveness of the phasing out of PFOS production in the 2000s. In contrast, ∑PFCAs concentrations in eggs were constant and low in the 1970s and 1980s, suggesting minimal environmental contamination, but residues subsequently increased significantly in both colonies until the end of the study. This increase appeared driven by rises in long chain compounds, namely the odd chain numbered PFTriDA and PFUnA. PFOA, had a very different temporal trend from the other dominant acids, with an earlier rise in concentrations followed by a decline in the last 15 years in Ailsa Craig; later temporal trends in Bass Rock eggs were unclear. Although eggs from both colonies contained relatively low concentrations of PFAS, the majority had PFOS residues that exceeded a suggested Predicted No Effect Concentration and ~ 10% of the eggs exceeded a suggested Lowest-Observable-Adverse-Effect.

Decabromodiphenylether trends in the European environment: Bird eggs, sewage sludge and surficial sediments.

Concern on relatively high levels and the potential bioaccumulation of decabromodiphenylether (BDE209) has led to a European 8-year monitoring program on trends in BDE209 concentrations in birds, sewage sludge and sediments from seven countries. BDE209 was analysed in four environmental matrices: sparrowhawk eggs (UK), glaucous gull eggs (Bear Island, Norway), sewage sludge (UK, Ireland and the Netherlands) and sediment (France, Germany, the Netherlands, UK and Ireland). BDE209 was detected in most of the glaucous gull and sparrow hawk eggs but neither increasing nor decreasing trends in these BDE209 levels were observed. An indication for debromination of BDE209 in sparrowhawk eggs was found. BDE209 concentrations in sediments ranged from very low ng/g (88 ng/g on an organic carbon (OC) basis) concentrations, in the rivers Elbe, Ems, Seine and the Outer Humber, to high µg/g (120 µg/g OC), in the Western Scheldt, Liverpool Bay and River Mersey. Apart from decreasing values in the Western Scheldt sediment no further decreases in BDE209 concentrations were observed over time, neither in sediment nor in sewage sludge showing that the voluntary emissions control program of the bromine industry only had a local effect. In contrast to the sewage sludge samples from the Netherlands (mean 355 ng/g dry weight (dw) or 1026 ng/g OC), the BDE209 concentrations in the UK increased at all sites from 2006 to 2011 (8092 ng/g dw or 22,367 ng/g OC). The BDE209 levels in several UK sediments and sewage sludge were still very high at the end of the program in 2012, most likely caused by frequent use of BDE209 in the textile industry. This may be indicative of the persistence of BDE209 and the limited degradation into lower brominated congeners in sediment, although it cannot be excluded that ongoing BDE209 emissions have played a role as well.

A review of constraints and solutions for collecting raptor samples and contextual data for a European Raptor Biomonitoring Facility.

The COST Action 'European Raptor Biomonitoring Facility' (ERBFacility) aims to develop pan-European raptor biomonitoring in support of better chemicals management in Europe, using raptors as sentinel species. This presents a significant challenge involving a range of constraints that must be identified and addressed. The aims of this study were to: (1) carry out a comprehensive review of the constraints that may limit the gathering in the field of raptor samples and contextual data, and assess their relative importance across Europe; and (2) identify and discuss possible solutions to the key constraints that were identified. We applied a participatory approach to identify constraints and to discuss feasible solutions. Thirty-one constraints were identified, which were divided into four categories: legal, methodological, spatial coverage, and skills constraints. To assess the importance of the constraints and their possible solutions, we collected information through scientific workshops and by distributing a questionnaire to stakeholders in all the countries involved in ERBFacility. We obtained 74 answers to the questionnaire, from 24 of the 39 COST participating countries. The most important constraints identified were related to the collection of complex contextual data about sources of contamination, and the low number of existing raptor population national/regional monitoring schemes and ecological studies that could provide raptor samples. Legal constraints, such as permits to allow the collection of invasive samples, and skills constraints, such as the lack of expertise to practice necropsies, were also highlighted. Here, we present solutions for all the constraints identified, thus suggesting the feasibility of establishing a long-term European Raptor Sampling Programme as a key element of the planned European Raptor Biomonitoring Facility.

Biological and anthropogenic predictors of metal concentration in the Eurasian otter, a sentinel of freshwater ecosystems.

Toxic metals have been linked to a range of adverse health effects in freshwater organisms. However, for higher vertebrates, there is little understanding of the large-scale drivers of exposure. We quantified toxic metal/semi-metal concentrations in a sentinel freshwater top predator, the Eurasian otter (Lutra lutra), across England and Wales, and determined how this varied with key natural and anthropogenic factors. We related liver concentrations in 278 otters that died between 2006 and 2017 to habitat biogeochemistry, proximity to point source contamination and to biological characteristics (length, sex, condition). Evidence for any positive association with putative anthropogenic sources (mining, human population, known discharges) was weak or lacking in nearly all cases, with the exception of a positive association between lead and human population density. Despite concerns that burgeoning use of nanosilver in consumer products might increase silver concentrations in waste waters, there was no increase over time. Spatial variation in soil/sediment pH, precipitation, and soil calcium oxide are indicated as significant predictors of metal concentrations in otters (higher cadmium and silver in areas with lower pH and higher rainfall, and higher chromium and lead in areas of lower calcium oxide). Liver chromium and nickel concentrations declined significantly over time (Cr 0.030 ± 1.2 to 0.015 ± 1.3 µg/g dry weight, Ni 0.0038 ± 1.2 to 0.00068 ± 1.5 µg/g, between 2006-2009 and 2014-2017), but other metals showed no temporal change. Biotic associations were important, with age related accumulation indicated for mercury and cadmium (as well as interactions with body condition). Our results suggest that larger-scale geochemical and hydrological processes are important in determining metal exposure in otters, and we provide an indication of risk factors that may be of relevance for freshwater vertebrates in other countries with well-developed water pollution management.

Concentration and origin of lead (Pb) in liver and bone of Eurasian buzzards (Buteo buteo) in the United Kingdom.

Ingestion of lead (Pb) derived from ammunition used in the hunting of game animals is recognised to be a significant potential source of Pb exposure of wild birds, including birds of prey. However, there are only limited data for birds of prey in Europe regarding tissue concentrations and origins of Pb. Eurasian buzzards (Buteo buteo) found dead in the United Kingdom during an 11-year period were collected and the concentrations of Pb in the liver and femur were measured. Concentrations in the liver consistent with acute exposure to Pb were found in 2.7% of birds and concentration in the femur consistent with exposure to lethal levels were found in 4.0% of individuals. Pb concentration in the femur showed no evidence of consistent variation among or within years, but was greater for old than for young birds. The Pb concentration in the liver showed no effect of the birds' age, but varied markedly among years and showed a consistent tendency to increase substantially within years throughout the UK hunting season for gamebirds. The resemblance of the stable isotope composition of Pb from buzzard livers to that of Pb from the types of shotgun ammunition most widely-used in the UK increased markedly with increasing Pb concentration in the liver. Stable isotope results were consistent with 57% of the mass of Pb in livers of all of the buzzards sampled being derived from shotgun pellets, with this proportion being 89% for the birds with concentrations indicating acute exposure to Pb. Hence, most of the Pb acquired by Eurasian buzzards which have liver concentrations likely to be associated with lethal and sublethal effects is probably obtained when they prey upon or scavenge gamebirds and mammals shot using Pb shotgun pellets.

Key principles and operational practices for improved nanotechnology environmental exposure assessment.

Nanotechnology is identified as a key enabling technology due to its potential to contribute to economic growth and societal well-being across industrial sectors. Sustainable nanotechnology requires a scientifically based and proportionate risk governance structure to support innovation, including a robust framework for environmental risk assessment (ERA) that ideally builds on methods established for conventional chemicals to ensure alignment and avoid duplication. Exposure assessment developed as a tiered approach is equally beneficial to nano-specific ERA as for other classes of chemicals. Here we present the developing knowledge, practical considerations and key principles need to support exposure assessment for engineered nanomaterials for regulatory and research applications.

Second generation anticoagulant rodenticides in tawny owls (Strix aluco) from Great Britain.

Secondary exposure of vertebrate predators to second generation anticoagulant rodenticides (SGARs) is widespread in Britain. Tawny owl (Strix aluco) populations in the UK are thought to have declined since the 1970s, when SGARs were first introduced, and these compounds may have contributed to any decline in owl numbers. Our aims were to conduct the first systematic survey of SGAR exposure in tawny owls and ascertain whether there had been a change in the proportion of exposed birds that was concurrent with the decline in the population. Liver difenacoum, bromadiolone, flocoumafen and brodifacoum concentrations in British tawny owls from two periods (1990-1993 and 2003-2005) were quantified. In total, some 20% of birds contained detectable residues of one or more SGAR. The extent of exposure (% of birds exposed, magnitude of residues) to different SGARs did not change consistently between time periods. Of the raptors analysed to date in Britain, tawny owls had the lowest proportion of individuals that contained detectable liver residues and so appear to be the least vulnerable to exposure and/or assimilation of SGARs. We found no clear evidence to implicate SGARs as a major factor affecting tawny owl numbers in Britain between 1990 and 2005.

The Predatory Bird Monitoring Scheme: identifying chemical risks to top predators in Britain.

The Predatory Bird Monitoring Scheme (PBMS) is a long term (>40 y), UK-wide, exposure monitoring scheme that determines the concentration of selected pesticides and pollutants in the livers and eggs of predatory birds. This paper describes how the PBMS works, and in particular highlights some of the key scientific and policy drivers for monitoring contaminants in predatory birds and describes the specific aims, scope, and methods of the PBMS. We also present previously unpublished data that illustrates how the PBMS has been used to demonstrate the success of mitigation measures in reversing chemical-mediated impacts; identify and evaluate chemical threats to species of high conservation value; and finally to inform and refine monitoring methodologies. In addition, we discuss how such schemes can also address wider conservation needs.

The importance of the gut and its contents in prey as a source of cadmium to predators.

Many mammalian and avian predators consume the whole carcass of their small mammal prey. Thus, the contaminant load of the gut and its contents may be of toxicological significance, but pollutant loads in the gut of prey species are often not measured and their role in food-chain transfer of contaminants is largely unquantified. We investigated the importance of the gut and its contents in prey as a source of cadmium (Cd) to predators by measuring the Cd concentrations in the guts (including contents), in various body tissues, and in the intact bodies of wood mice (Apodemus sylvaticus). The mice had been fed for up to 365 d on either control, a medium-Cd (MCd) diet, or a high-Cd (HCd) diet that had nominal Cd concentrations of 0, 8, and 40 g/g wet weight, respectively. In wood mice fed contaminated diets, Cd was progressively bioaccumulated in all the body tissues that were analyzed. The gut (and contents) and the liver and kidneys (the critical organs) contained most of the accumulated Cd and were the only significant contributors to the total body burden (TBB). The contribution of the gut (and contents) to the TBB was greater in mice fed MCd diet than in those given HCd feed but declined with age (equivalent to exposure duration) in both groups as Cd in the critical organs increasingly became the most important source of TBB Cd. However, the gut and its contents contributed between 50 and 100% of the Cd TBB in mice fed for up to six months on MCd diet; this was the diet that most closely mimicked the estimated average Cd intake of wood mice on contaminated sites. The results of this study highlight the importance of including measures of Cd load in the gut when modeling food-chain transfer from small mammals to their predators.

Modeling cadmium dynamics in the guts and tissues of small mammals: dose implications for predators.

Predators that consume contaminated small-mammal prey are exposed to pollutants present in both the prey tissues and the prey gut. However, the contribution of the prey gut and its contents to the contaminant intake of the predator is often ignored. This study developed a two-compartment model of the dynamics of Cd in a model prey species, the wood mouse (Apodemus sylvaticus). The results from the Cd dynamics model were used to calibrate a revised predator dose estimation methodology that accounted for contaminant uptake by a predator from both prey tissues and guts. The dose to the predator predicted by the revised methodology was compared to that predicted by conventional dose calculations, which account for contaminants in prey tissues only. The conventional approach underestimated Cd dose to a model predator by a factor of 1.2- to 10-fold, depending on prey age and predator behavior. Thus, accounting for the contaminant burden in the prey gut as well as in the tissues can markedly increase dose estimates to predators, and this is likely to be true for most low-bioavailability pollutants. This study has major implications for ecological risk assessment involving food chain transfer of low-bioavailability xenobiotics to predators.

How much do PCB toxic equivalents account for PHAH toxicity in predatory birds?

Various diffuse polyhalogenated aromatic hydrocarbons (PHAHs) exert common toxicity through the aryl hydrocarbon receptor (AhR). Apex predators spatially and temporally integrate diffuse contamination and simultaneous exposure can cause additive toxicity. We investigated the extent to which PCBs, still amongst the most prevalent PHAHs accumulated by predators, accounted for total PHAH toxicity in raptors and fish eating birds from Britain. We analysed egg or liver extracts from six species and compared chemically determined ΣPCB-TEQs concentrations with total AhR-mediated toxicity determined using the chemical-activated luciferase gene expression bioassay (CALUX-TEQ). Dioxin-like PCB profiles in eggs and livers were dominated by congeners 118, 105 and 167. ΣPCB-TEQ and CALUX-TEQ concentrations were positively associated but not in a 1:1 relationship. ΣPCB-TEQ were broadly similar to CALUX-TEQ concentrations in eggs and livers with CALUX-TEQ concentrations >50-80 and 160-320 pg g(-1) lipid respectively, but were lower than CALUX-TEQ concentrations in less contaminated samples.

Comparisons of metabolic and physiological changes in rats following short term oral dosing with pesticides commonly found in food.

¹H Nuclear Magnetic Resonance spectroscopy has been used to profile urinary metabolites in male Fischer F344 rats in order to assess the metabolic changes induced by oral exposure to two benzimidazole fungicides (carbendazim and thiabendazole) and two bipyridyllium herbicides (chlormequat and mepiquat). Exposure levels were selected to be lower than those expected to cause overt signs of toxicity. We then compared the sensitivity of the metabolomics approach to more traditional methods of toxicity assessment such as the measurement of growth and organ weights. Separate, acute exposure experiments were conducted for each pesticide to identify potential metabolic markers of exposure across four doses (and a control). Growth, organ weights and feeding/drinking rates were not significantly affected by any compounds at any dose levels tested. In contrast, metabolic responses were detected within 8 and 24h for chlormequat and mepiquat, and after 24h for carbendazim and thiabendazole. These results demonstrate the potential for the use of metabolomics in food toxicity testing.

Decabromodiphenylether and hexabromocyclododecane in wild birds from the United Kingdom, Sweden and The Netherlands: Screening and time trends.

The brominated flame retardant decabromodiphenylether (DBDE) was analysed in wild birds to identify the most suitable species for monitoring time trends in DBDE contamination. This information was later used for the design of a 10-year trend study on DBDE in the European Union. DBDE was measured in muscle tissue, liver, and eggs from 10 terrestrial and four aquatic bird species. DBDE was detected in 47% of the terrestrial bird samples (nine species) and in 9% of the aquatic bird samples (six species). Peregrine falcon and sparrowhawk specimens were selected as most suitable species to determine temporal trends of DBDE. For sparrowhawks, no significant change in DBDE concentrations between 1973 and 2001 was found, although in later years more DBDE concentrations were above the detection limit. Peak DBDE levels measured in peregrines in 1995, were followed by a decline in concentrations until 2001. The same species were used for a trend study on hexabromocylcododecane (HBCD). Twenty-four percent of peregrine falcon eggs and 12% of sparrowhawk muscle samples demonstrated measurable HBCD residues. Three diastereomers of HBCD were analysed and the α-diastereomer was the predominant one in most samples. No clear time trends were observed for HBCD in either species. This study demonstrated that these DBDE and HBCD are bioavailable to birds of Northern Europe, although bioaccumulation seems to occur to a limited extent.

Long-term trends in mercury and PCB congener concentrations in gannet (Morus bassanus) eggs in Britain.

Gannet (Morus bassanus) eggs from Bass Rock (North Sea) and Ailsa Craig (eastern Atlantic) were monitored for PCB congeners (1990-2004) and total mercury (1974-2004). Congener profiles for both colonies were dominated by PCBs 153, 138, 180, 118 and 170. All declined in concentration at Ailsa Craig but some (153, 170, 180) remained stable or increased slightly at Bass Rock. Egg congener concentrations at Bass Rock were typically 10-fold higher than at Ailsa Craig by 2002, and Principal Component Analysis indicated that colony differences were driven by the dominant congeners. Egg mercury concentrations were significantly lower at Bass Rock than at Ailsa Craig and temporal trends differed, there being a significant decline at Ailsa Craig but a marginal increase at Bass Rock. Our results suggest there may be differences in contamination between the eastern Atlantic and North Sea and/or there are colony differences in prey selection and associated contaminant loads.

Polybrominated diphenyl ether-associated alterations in cell biochemistry as determined by attenuated total reflection Fourier-transform infrared spectroscopy: a comparison with DNA-reactive and/or endocrine-disrupting agents.

Whether polybrominated diphenyl ethers (PBDEs) induce effects in target cells is increasingly important given that their environmental burdens are rising. Attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy can be used to biochemically signature cells based on the notion that a detailed "biochemical-cell fingerprint" in the form of an infrared (IR) spectrum is derived. By employing subsequent computational approaches such as principal component analysis (PCA) and/or linear discriminant analysis (LDA), data reduction is achieved to allow for the identification of wavenumber-related biomarkers of effect Clustering of similar spectra (or scores) away from dissimilar ones highlights the variance responsible for discriminating classes. Discriminating biomarkers might include protein conformational changes, structural alterations to DNA/RNA, glycogen content, or protein phosphorylation. Employing this approach, we investigated in MCF-7 cells the dose-related effects of PBDEs (congeners 47, 153, 183, and 209), benzo[a]pyrene (B[a]P), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PHIP), 17beta-Oestradiol (E2), or lindane (gamma-hexachlorocyclohexane). Cultures concentrated in G0/G1- or S-phases were treated for 24 h. Following treatment MCF-7 cells were fixed and applied to IR reflective Low-E windows for interrogation using ATR-FTIR spectroscopy. At concentrations as low as 10(-12) M in culture, significant separation (P < or = 0.05) of PBDE-treated and vehicle control cell populations was noted. In some cases this was associated with alterations in lipid or the secondary structure of proteins; with DNA-reactive compounds (e.g., B[a]P), variance was primarily noted in the DNA/RNA region. This study points to a novel nondestructive approach capable of identifying contaminant effects at environmental concentrations in target cells.

Concentrations of polycyclic aromatic hydrocarbons (PAHs) in the eggs of predatory birds in Britain.

The eggs of gannets from two Scottish colonies (Ailsa Craig, Bass Rock) of golden eagles from the Hebrides and Highlands and of merlin eggs from the Scottish borders were analyzed for 52 PAHs, including 2-7 ring parent and alkylated PAHs. Phenanthrene was the most abundant PAH in gannet eggs from Ailsa Craig, and methylnaphthalenes predominated in the eggs from other locations and species. Most PAHs were detected in eggs but none were at likely embryotoxic concentrations. The sum concentrations for all the PAHs analyzed (3.1-5.7 ng g(-1) wet wt.) and for the U.S. EPA 16 priority PAHs (2.0-4.3 ng g(-1) wet wt.) did not differ significantly between species or locations. This uniform, low-level accumulation suggests background exposure to diffuse sources. PCA indicated that 3 ring parent and alkylated PAHs predominated in the eggs of merlins and gannets from Ailsa Craig and Hebridean golden eagles; other eggs had a more mixed profile. Source signature diagnostics largely suggested a petrogenic origin for the PAHs in the merlin eggs that we analyzed but otherwise gave equivocal results and further work is needed to determine which diagnostics can be successfully applied to PAHs in eggs.