D. Osborn

One Hundred Questions of Importance to the Conservation of Global Biological Diversity

We identified 100 scientific questions that, if answered, would have the greatest impact on conservation practice and policy. Representatives from 21 international organizations, regional sections and working groups of the Society for Conservation Biology, and 12 academics, from all continents except Antarctica, compiled 2291 questions of relevance to conservation of biological diversity worldwide. The questions were gathered from 761 individuals through workshops, email requests, and discussions. Voting by email to short-list questions, followed by a 2-day workshop, was used to derive the final list of 100 questions. Most of the final questions were derived through a process of modification and combination as the workshop progressed. The questions are divided into 12 sections: ecosystem functions and services, climate change, technological change, protected areas, ecosystem management and restoration, terrestrial ecosystems, marine ecosystems, freshwater ecosystems, species management, organizational systems and processes, societal context and change, and impacts of conservation interventions. We anticipate that these questions will help identify new directions for researchers and assist funders in directing funds.

Polycyclic aromatic hydrocarbon (PAH) dispersion and deposition to vegetation and soil following a large scale chemical fire

Polycyclic aromatic hydrocarbons (PAHs) were determined in soil and vegetation following a large scale chemical fire involving 10,000 ton of polypropylene. In comparison with sites outside the plume from the fire, PAH concentrations were elevated in grass shoots (by up to 70-fold) and in soil (by up to 370-fold). The pattern of PAH dispersion under the plume was dependent on the physical-chemical properties of individual PAHs. The lighter, least hydrophobic PAHs were dispersed into the environment at greater distances than heavier, more hydrophobic PAHs. At the most distant sampling point (4.5 km) under the plume, the low molecular weight PAHs were still considerably elevated in vegetation samples compared to control sites. Dispersion appeared to be regulated by the compounds partitioning between the vapour and particulate phase, with dry particulate deposition occurring closer to the fire source than gaseous deposition. For all PAHs, the fire resulted in greater contamination of soils compared to grasses, with the relative ratio of plant/soil contamination decreasing as hydrophobicity increased.

High levels of cadmium in Atlantic seabirds and sea-skaters

THE possibility that cadmium may be an environmental pollutant has caused concern because it has toxic effects on many animal species including man, rat, rabbit, chicken, quail, and pigeon1–4. The occurrence of cadmium in wildlife is not unusual and samples of marine species taken from around the British coast contain low levels (less than 0.5 mg per kg in fish and less than 2 mg per kg in most shellfish); however, levels of up to 93 mg per kg were present in limpets, Patellasp., collected from areas of natural mineralisation or industrial sources of cadmium5. Seabirds generally contain higher residues than marine invertebrates. For example, Anderlini et al.6 examined the amounts of nine metals in the livers of seven Antarctic and Pacific seabird species and noted that the maximum mean cadmium residue occurred in 10 ashy petrels Oceanodroma homochroa collected from their breeding grounds in California (mean residue was 53.2±20.5 mg per kg). Parslow et al.7 found up to 22.3 mg per kg dry wt cadmium in livers of eight puffins Fratercula arctica collected live from various British breeding colonies. It was suggested that pelagic birds might contain higher residues than coastal living species because a single fulmar Fulmarus glacialis examined by them contained 159 mg per kg cadmium. Anderlini et al. concluded that there was a correlation between increased concentration of cadmium in birds and exposure to industrial influences. Parslow et al. did not attempt to explain the residues they found in puffins, but Bourne8 concluded that individual birds were becoming increasingly contaminated through feeding near areas of local pollution around the British coast. It is debatable whether cadmium should be regarded as a pollutant to seabirds. We report here the occurrence of cadmium residues in the tissues of apparently healthy, breeding seabirds which are considerably higher than any previously found. More important, we conclude that the cadmium has a natural rather than an industrial origin, since we have found high cadmium residues in a marine insect, Halobates, which is widely distributed in tropical regions far from sources of industrial cadmium. This insect is only one example of the sources from which birds obtain their cadmium residues.

An NMR‐based metabonomic approach to the investigation of coelomic fluid biochemistry in earthworms under toxic stress

The endogenous metabolites of the coelomic fluid of the earthworm Eisenia veneta were characterised using high‐resolution one‐dimensional and two‐dimensional 1H nuclear magnetic resonance spectroscopy. Signals from common organic acids, such as acetate, fumarate, malonate, malate, formate, and succinate, were identified together with adenosine and nicotinamide mononucleotide. The potential use of this information as a baseline dataset for future toxicological or physiological studies was demonstrated by a metabonomic analysis: a series of earthworms were dosed with the model compound 3‐fluoro‐4‐nitrophenol, and toxic effects followed by multivariate analysis of the spectral data of the coelomic fluid. Relative concentrations of acetate and malonate were decreased in the dosed worms compared to the controls.

Comparative tissue distribution of mercury, cadmium and zinc in three species of pelagic seabirds

Abstract 1. The levels of mercury cadmium and zinc were measured in tissues of puffin ( Fratercula arctica ) fulmar ( Fulmarus glacialis ) and Manx shearwater ( Puffinus puffinus ). 2. The highest levels of zinc and cadmium (up to 480mg/kg Cd, dry wt) were found in kidney, liver, pancreas, gonad, and intestine. 3. Substantial quantities of mercury-mostly methyl mercury-were found in the liver of the fulmar nd Manx shearwater (up to 45.0 mg/kg Hg, dry wt). 4. Puffin feathers contained more mercury (7.94 mg/kg Hg, dry wt) than the liver and kidney. 5. The possibility that, since these birds were breeding they were not suffering any adverse effects of the metals, is discussed.

Earthworm species of the genus Eisenia can be phenotypically differentiated by metabolic profiling

The universality of low molecular weight metabolites allows rapid and straightforward investigation of the biochemistry of genetically uncharacterised species. Thus ex vivo metabolic profiling in combination with multivariate data analysis (metabonomics) offers great potential in comparative biology. Here we present the first use of high resolution nuclear magnetic resonance (NMR) spectroscopy to distinguish closely related animal species via their metabolic phenotype (metabotype). We have profiled the three Eisenia (Oligochaeta, Lumbricidae) species Eisenia fetida, Eisenia andrei and Eisenia veneta using tissue extracts and coelomic fluid analysis. The low molecular weight biochemical profiles of tissue extracts were highly conserved for all three species, with E. fetida and E. andrei being more similar to each other than to E. veneta. However the metabolic profiles of the coelomic fluid of the different species were highly distinctive – the NMR spectra allowed unequivocal identification of species. Multivariate statistics were also used to quantify these spectral differences and to enable simplified graphical visualisation of species similarity. These results show that two morphologically undistinguishable species (E. fetida and E. andrei) differ markedly in their biochemical profiles despite apparently occupying the same ecological niche, and indicate that metabolic phenotype profiling can be used as a powerful functional genomics tool.

SEASONAL-CHANGES IN THE FAT, PROTEIN AND METAL CONTENT OF THE LIVER OF THE STARLING STURNUS-VULGARIS

Abstract In the starling liver there are seasonal variations in the concentrations of zinc, copper, iron, cadmium and mercury. Some of these variations may be related to seasonal changes in the fat and protein content of the liver. It is suggested that an understanding of the significance of the levels of toxic metals in any animal will depend upon an understanding of the seasonal changes in the protein composition of tissues.

The initial pathogenesis of cadmium induced renal toxicity

The novel application of magic angle spinning 1H NMR spectroscopy, coupled with pattern recognition techniques, has identified biochemical changes in lipid and glutamate metabolism that precede classical nephrotoxicity. These changes occurred in the bank vole (Clethrionomys glareolus) after chronic dosing, at a low level of exposure and at a renal Cd2+ concentration (8.4 μg/g dry wt) that was nearly two orders of magnitude below the WHO critical organ concentration (200 μg/g wet wt). These early stage effects of Cd2+ on the biochemistry of renal tissue may reflect adaptation mechanisms to the toxic insult or the preliminary stages of the toxicological cascade.

Chlorobenzenes in rivers draining industrial catchments.

Eleven chlorobenzenes (out of a total of 12 in the congener series) were monitored weekly on four industrialized rivers (Aire, Calder, Don and Trent) of the Southern Humber Catchment in whole water samples. 1,2- and 1,4-dichlorobenzene were present at relatively high levels on both the Aire and Calder, having mean concentrations of approximately 30 ng/l. They were both at lower concentrations on the Don and Trent, although the 1,4-isomer dominated. All other chlorobenzenes monitored were routinely found on all the rivers, with the exception of hexachlorobenzene, which was only regularly detected on the Trent. Again, the rivers fell into two classes with respect to their total chlorobenzene concentrations, with the Aire and Calder being more polluted. The higher levels of chlorobenzenes (excluding hexachlorobenzene which was used widely as a agricultural pesticide) on the Aire and Calder, and the dominance of the 1,4-dichlorobenzene congener (accounting for 60-70% of sigma chlorobenzenes) on the Don and Trent, indicated that the Aire and Calder were predominately contaminated with chlorobenzenes through industrial sources, while the Don and Trent were mainly contaminated through domestic sources (1,4-dichlorobenzene is widely used as a toilet deodorant). 1,4-Dichlorobenzene dominated flux, with the Aire, Don and Trent exporting 52.5 kg/year into the Humber estuary, followed by the 1,2-dichlorobenzene at 38.8 kg/year. Sigma chlorobenzenes exported to the Humber was 133 kg/year. This is the first study to calculate chlorobenzene fluxes to the North Sea from a UK catchment.

An NMR-based metabonomic investigation of the toxic effects of 3-trifluoromethyl-aniline on the earthworm Eisenia veneta

(1)H NMR spectra of earthworms Eisenia veneta treated with 3-trifluoromethyl-aniline in a 72-h contact filter paper test have been analysed using pattern recognition techniques to determine the biochemical response. Various strategies for data reduction of the metabolite profile, and illustration by principal components analysis are applied and discussed. The use of mean principal components plots in simplifying group data representation and highlighting the dose-response function is demonstrated. Hierarchical cluster analysis, and cluster significance analysis of the principal components were also used to examine the relative distribution of dose groups. Identification and assignment of metabolite responses to toxicity were found via correlation coefficient-shift plots. As measured by the correlation coefficients alanine was the most significant metabolite, but increased levels of other amino acids such as glycine and asparagine were also observed. Further, elevated levels of glucose, and the citric acid cycle intermediates citrate and succinate were noted as potential biomarkers of toxicity. This work provides a basis for examining the biochemical response of invertebrates to toxins. This should provide a framework to examine toxicity effects of other halogenated aromatic pollutants to earthworms used as environmental monitors.1H NMR spectra of earthworms Eisenia veneta treated with 3-trifluoromethyl-aniline in a 72-h contact filter paper test have been analysed using pattern recognition techniques to determine the biochemical response. Various strategies for data reduction of the metabolite profile, and illustration by principal components analysis are applied and discussed. The use of mean principal components plots in simplifying group data representation and highlighting the dose-response function is demonstrated. Hierarchical cluster analysis, and cluster significance analysis of the principal components were also used to examine the relative distribution of dose groups. Identification and assignment of metabolite responses to toxicity were found via correlation coefficient-shift plots. As measured by the correlation coefficients alanine was the most significant metabolite, but increased levels of other amino acids such as glycine and asparagine were also observed. Further, elevated levels of glucose, and the citric acid cycle intermediates citrate and succinate were noted as potential biomarkers of toxicity. This work provides a basis for examining the biochemical response of invertebrates to toxins. This should provide a framework to examine toxicity effects of other halogenated aromatic pollutants to earthworms used as environmental monitors.