A. R. Brown

Genetic variation, inbreeding and chemical exposure—combined effects in wildlife and critical considerations for ecotoxicology

Exposure to environmental chemicals can have negative consequences for wildlife and even cause localized population extinctions. Resistance to chemical stress, however, can evolve and the mechanisms include desensitized target sites, reduced chemical uptake and increased metabolic detoxification and sequestration. Chemical resistance in wildlife populations can also arise independently of exposure and may be spread by gene flow between populations. Inbreeding—matings between closely related individuals—can have negative fitness consequences for natural populations, and there is evidence of inbreeding depression in many wildlife populations. In some cases, reduced fitness in inbred populations has been shown to be exacerbated under chemical stress. In chemical testing, both inbred and outbred laboratory animals are used and for human safety assessments, iso-genic strains (virtual clones) of mice and rats are often employed that reduce response variation, the number of animals used and associated costs. In contrast, for environmental risk assessment, strains of animals are often used that have been selectively bred to maintain heterozygosity, with the assumption that they are better able to predict adverse effects in wild, genetically variable, animals. This may not necessarily be the case however, as one outbred strain may not be representative of another or of a wild population. In this paper, we critically discuss relationships between genetic variation, inbreeding and chemical effects with the intention of seeking to support more effective chemical testing for the protection of wildlife.

Assessing variation in the potential susceptibility of fish to pharmaceuticals, considering evolutionary differences in their physiology and ecology

Fish represent the planets most diverse group of vertebrates and they can be exposed to a wide range of pharmaceuticals. For practical reasons, extrapolation of pharmaceutical effects from ‘model’ species to other fish species is adopted in risk assessment. Here, we critically assess this approach. First, we show that between 65% and 86% of human drug targets are evolutionarily conserved in 12 diverse fish species. Focusing on nuclear steroid hormone receptors, we further show that the sequence of the ligand binding domain that plays a key role in drug potency is highly conserved, but there is variation between species. This variation for the oestrogen receptor, however, does not obviously account for observed differences in receptor activation. Taking the synthetic oestrogen ethinyloestradiol as a test case, and using life-table-response experiments, we demonstrate significant reductions in population growth in fathead minnow and medaka, but not zebrafish, for environmentally relevant exposures. This finding contrasts with zebrafish being ranked as more ecologically susceptible, according to two independent life-history analyses. We conclude that while most drug targets are conserved in fish, evolutionary divergence in drug-target activation, physiology, behaviour and ecological life history make it difficult to predict population-level effects. This justifies the conventional use of at least a 10× assessment factor in pharmaceutical risk assessment, to account for differences in species susceptibility.

Qualitative and Quantitative Histomorphologic Assessment of Fathead Minnow Pimephales promelas Gonads as an Endpoint for Evaluating Endocrine-Active Compounds: A Pilot Methodology Study

Although histopathology is routinely employed as a tool for the detection and assessment of xenobiotic-mediated effects in mammals, it is less frequently applied to fish. In part, this is due to a lack of method standardization regarding study design, tissue preservation, tissue sectioning, histopathological evaluation, reporting, and statistical analysis. The objectives of the present study were: (1) to test and refine a method for the microsurgical excision of fathead minnow (FHM) Pimephales promelas gonads for the purpose of histopathologic examination; (2) to determine the optimal combination of fixation and embedding procedures for the histopathologic and morphometric analysis of FHM gonads following exposure to a known estrogenic compound, 17β-estradiol (E2); and (3) to provide a method for the categorization and quantification of cell types in FHM gonads by manually counting cells in digitized images using image analysis software. The light microscopic evaluation of individual gametogenic cells was greatly facilitated by specimen preparation techniques that included the excision of gonads via microdissection and by optimized fixation and embedding procedures.

Contaminants in fine sediments and their consequences for biota of the Severn Estuary

When the first MPB special issue was published 25 years ago it was suggested that high body burdens of metals and selected organic pollutants in the Severn Estuary were the result of anthropogenic loadings from a variety of sources. The objective of this synopsis is to illustrate recent trends for contaminants (metals, PAHs, PCBs) in sediments and benthic biota and to consider the evidence for improved environmental quality over the last quarter of a century. Contaminants in sediments and sediment-dwelling fauna such as Hediste(=Nereis)diversicolor are, generally, evenly distributed over the estuary - which is the consequence of extensive re-suspension and redistribution of fine sediment by strong tidal currents. Such dispersal tends to mask the influences of individual discharges and physical characteristics are considered to be the major drivers affecting biodiversity in the Severn Estuary, often overshadowing contaminant concerns. Following the closure of major industries and the introduction of stricter pollution control, many inputs have ceased or been reduced and there are indications that environmental concentrations are now lower. Bioaccumulation of most contaminants has declined accordingly (with the possible exception of Cr). Intuitively, better environmental quality should be linked to ecological improvements. However, due to the dynamic nature of the system (and a lack of biological-effects data) it is difficult to establish direct relationships between inputs, body burdens and biological/ecological consequence. Uniquely, the long-term integrated monitoring program of AstraZeneca (Avonmouth) indicates that recovery of faunal diversity and abundance has occurred in mid-sections of the estuary in recent years implying that contaminants have indeed been a forcing feature for Severn biota. In this context, we highlight contaminant issues and biogeochemical changes which may need to be addressed in connection with the development of proposals for tidal energy schemes.

Interactive effects of inbreeding and endocrine disruption on reproduction in a model laboratory fish

Inbreeding depression is expected to be more severe in stressful environments. However, the extent to which inbreeding affects the vulnerability of populations to environmental stressors, such as chemical exposure, remains unresolved. Here we report on the combined impacts of inbreeding and exposure to an endocrine disrupting chemical (the fungicide clotrimazole) on zebrafish (Danio rerio). We show that whilst inbreeding can negatively affect reproductive traits, not all traits are affected equally. Inbreeding depression frequently only became apparent when fish were additionally stressed by chemical exposure. Embryo viability was significantly reduced in inbred exposed fish and there was a tendency for inbred males to sire fewer offspring when in direct competition with outbred individuals. Levels of plasma 11‐ketotestosterone, a key male sex hormone, showed substantial inbreeding depression that was unaffected by addition of the fungicide. In contrast, there was no effect of inbreeding or clotrimazole exposure on egg production. Overall, our data provide evidence that stress may amplify the effects of inbreeding on key reproductive traits, particularly those associated with male fitness. This may have important implications when considering the consequences of exposure to chemical pollutants on the fitness of wild populations.

Effects of VOCs on herbaceous plants in an open-top chamber experiment

A selection of herbaceous plants representing the ground flora around a typical chemical installation in the UK was exposed continuously for 7 weeks to a mixture of six VOCs (acetone, acetonitrile, dichloromethane, ethanol, methyl t-butyl ether and toluene) in open-top chambers. Exposure concentrations were based on predictions of atmospheric dispersion from a single source, at a distance of approximately 2 km. The effects of continuous exposure, representing a worst-case, were measured in terms of uncontrolled water loss from leaves, leaf wettability, chlorophyll content and fluorescence, dry matter production and detailed observations of changes in plant growth and phenology. There were significant effects of VOC exposure on seed production, leaf water content and photosynthetic efficiency in some plant species. Such effects may be detectable in vegetation close to major industrial point sources of VOCs, or as a result of an accidental release of material during manufacture or transport. Some of the species tested e.g. birdsfoot trefoil (Lotus corniculatus L.) seem to be promising as potential bioindicators for VOCs, but there may be other even more sensitive species waiting to be discovered. However, the most obvious and conveniently measured response to VOCexposure in the birdsfoot trefoil (premature senescence i.e. advanced timing of seed pod production) could easily be confused in the field with climatic influences. It is also uncertain at this stage whether any of the effects observed would lead to longer term ecological changes in natural plant communities, through biased competition between sensitive and more tolerant species.

‘I Went Disguised in It’: Re-evaluating Seamus Heaney’s Stations

In this essay, Brown offers a re-evaluation of Seamus Heaney’s 1973 prose poem collection, Stations, which has been criticised as both a ‘poor relation’ to the poet’s own lineated verse, as well as for the heroic imagery and language utilised. By discussing the generic conventions of the ‘anecdotal tradition’ and its relation to the prose poem, Brown refutes the earlier criticism and shows Heaney’s Stations to be exemplars of the form, as well as of Wordsworthian ‘spots of time’. Brown also shows Stations to be knowledgeably conversant with notions of ‘the Romantic Fragment’, as well as with more postmodern conceptions of the prose poem sentence as an emergent, generative syntactical unit. Brown also discusses the influence of Stations on subsequent generations of British prose poets.