Lis Bach

The use and misuse of biomarkers in ecotoxicology

Substantial efforts have been devoted to developing and applying biomarkers for use in ecotoxicology. These efforts have resulted partly from a desire for early warning indicators that respond before measurable effects on individuals and populations occur and partly as an aid to identifying the causes of observed population- and community-level effects. Whereas older biomarkers focused on measures of organism physiology or biochemistry, advances in molecular biology are extending the biomarker philosophy to the level of the genes (i.e., ecotoxicogenomics). However, the extent to which biomarkers are able to provide unambiguous and ecologically relevant indicators of exposure to or effects of toxicants remains highly controversial. In the present paper, we briefly discuss the application of biomarkers in ecotoxicology and ecological risk assessment, and we provide examples of how they have been applied. We conclude that although biomarkers can be helpful for gaining insight regarding the mechanisms causing observed effects of chemicals on whole-organism performance and may, in some cases, provide useful indicators of exposure, individual biomarker responses should not be expected to provide useful predictions of relevant ecological effects--and probably not even predictions of whole-organism effects. Suites of biomarkers are only likely to provide increased predictability if they can be used in a comprehensive mechanistic model that integrates them into a measure of fitness. Until this can be achieved, biomarkers may be useful for hypothesis generation in carefully controlled experiments. However, because the aims of environmental monitoring and ecological risk assessment are to detect and/or predict adverse chemical impacts on populations, communities, and ecosystems, we should be focusing our efforts on improving methods to do this directly. This will involve developing and testing models of appropriate complexity that can describe real-world systems at multiple scales.

Measuring bioavailable metals using diffusive gradients in thin films (DGT) and transplanted seaweed (Fucus vesiculosus), blue mussels (Mytilus edulis) and sea snails (Littorina saxatilis) suspended from monitoring buoys near a former lead-zinc mine in West Greenland.

Measuring loads of bioavailable metals is important for environmental assessment near mines and other industrial sources. In this study, a setup of monitoring buoys was tested to assess loads of bioavailable metals near a former Pb-Zn mine in West Greenland using transplanted seaweed, mussels and sea snails. In addition, passive DGT samplers were installed. After a 9-day deployment period, concentrations of especially Pb, Zn and Fe in the species were all markedly elevated at the monitoring sites closest to the mine. Lead concentrations in all three species and the DGT-Pb results showed a significant linear correlation. Zinc and Fe concentrations were less correlated indicating that the mechanisms for Zn and Fe accumulation in the three species are more complex. The results show that there is still a significant load of metals from the mine and that such buoys can be an adequate method to assess present loads of bioavailable metals.

Local contamination in relation to population genetic diversity and resilience of an arctic marine amphipod.

The objective of this study was to investigate whether populations inhabiting a contaminated environment are affected in terms of decreased genetic diversity due to selection of tolerant genotypes and if such effect carries a cost. Marine arctic amphipod populations (Orchomenella pinguis) were collected from sites within a contaminated fjord, as well as from sites outside the fjord on the west-coast of Greenland over three years (2006-2008). Impacts on genetic diversity, effects on resilience such as development of tolerance and cost were determined. AFLP-analysis was used to explore within and between population genetic diversity, and exposure studies were performed where the populations were subjected to known and unknown stressors to assess resilience. Populations collected at three contaminated sites all had reduced genetic diversity in 2007 compared to populations outside the fjord. This pattern was different in 2008 as all contaminated site populations increased in diversity, whereas a decrease in diversity occurred at the outer sites. However, tolerance, but even more so, cost, was related to contamination exposure in 2008, in spite of the shift in genetic diversity. We suggest that contamination rapidly induces effects that can be captured as tolerance and associated cost, whereas effects on genetic diversity can be difficult to separate from recent migration events that dilute eventual decreases in diversity due to contamination pressure. As long as impacted populations can be influenced by migration events that increase the genetic diversity and add health to an affected population, populations in contaminated areas may have enhanced probability of survival.

Evaluation of the use of common sculpin (Myoxocephalus scorpius) organ histology as bioindicator for element exposure in the fjord of the mining area Maarmorilik, West Greenland

The former Black Angel lead-zinc mine in Maarmorilik, West Greenland, is a historic example of how mining activity may result in a significant impact on the surrounding fjord system in terms of elevated concentrations of especially lead (Pb) and zinc (Zn) in seawater, sediments and surrounding biota. In order to shed light on the present contamination and possible effects in the fjord we initiated a range of studies including a pilot study on gill and liver morphology of common sculpins (Myoxocephalus scorpius) around Maarmorilik. Sculpins were caught and sampled at five different stations known to represent a gradient of Pb concentrations. Fish livers from all specimens were analyzed for relevant elements in the area: Fe, Zn, As, Cu, Se, Cd, Pb, Ag, Hg, Co and Ni. Lead, As and Hg showed significant differences among the five stations. For 20% of the sculpins, Hg concentrations were in the range of lowest observed effect dose (LOED) of 0.1-0.5 μg/g ww for toxic threshold on reproduction and subclinical endpoints. Likewise LOEDs for tissue lesions, LOEDs for biochemistry, growth, survival and reproduction were exceeded for Cd (0.42-1.8 μg/g ww) and for As (11.6 μg/g ww) in 28% and 85% of the sculpins, respectively. Similar to this, the no observed effect dose (NOED) for biochemistry was exceeded for Pb (0.32 μg/g ww) and for growth, mortality and reproduction for Zn (60-68 μg/g ww) in 33% and 24% of the sculpins, respectively. For all sculpins, females were significantly larger than males and for five of the elements (Fe, Co, Ni, Cu, Se) females had higher concentrations. The chronic lesions observed in liver (mononuclear cell infiltrates, necrosis, vacuolar hepatocytes, portal fibrosis, bile duct hyperplasia, active melanomacrophage centers) and gills (fusion and edema of secondary lamellae, laminar telangiectasis, mononuclear cell infiltrates, blebs) were similar to those in the literature studies for both wild and laboratory exposed sculpins and other fish species carrying similar or higher Hg concentrations. Ignoring sex and size, specimens with hepatic cell infiltrates had the highest concentrations of most elements, a relation that was also found for gill telangiectasis and Hg (all p < 0.05). When controlling for sex and size, the prevalence of vacuolar hepatocytes and endoparasites was significantly highest at the three most contaminated stations and similar differences were found for liver necrosis. We suggest that beside exposure to mining-related elements, other environmental factors, such as parasites, might be co-factors in the development of the observed liver and gill lesions. Therefore, sculpin liver and gill pathology are likely to be suitable health indicators when biomonitoring gradients of mining and other element related activity effects; while a larger study is required to fully evaluate the relationships.

The amphipod Orchomenella pinguis — A potential bioindicator for contamination in the Arctic

Indigenous organisms can be used as bioindicators for effects of contaminants, but no such bioindicator has been established for Arctic areas. Orchomenella pinguis is a benthic amphipod, ubiquitous in the Arctic and can be found in high numbers. We collected O. pinguis at sites with different contamination levels. Population characteristics (body length distribution, average dry weight and amphipod organic content) were related to sediment contaminant concentrations, in order to identify suitable endpoints for using this species as a bioindicator. We show that O. pinguis was prevalent in both clean and contaminated areas, easy to sample and that its population characteristics could be linked to both contamination and sediment organic content. We suggest that O. pinguis is a suitable bioindicator for the Arctic, but that endpoints such as reproductive effects and phenotypic and genotypic responses are needed together with population characteristics to assess impacts of contamination.

A simple method to reduce the risk of cadmium exposure from consumption of Iceland scallops (Chlamys islandica) fished in Greenland

This paper studied the levels and organ distribution of the toxic heavy metal cadmium in scallops from unpolluted Greenlandic waters. The scallops had an average cadmium concentration of 2.93 ± 0.94 μg/g wet weight in the total soft tissues and no concentration dependent effect was found for gender or size (both p>0.05). The kidney was the primary organ for cadmium accumulation with a mean of 226.2 ± 111.7 μg/g wet weight, and despite the small weight of the kidney, it appeared as the principal contributor of cadmium with 92% of the total cadmium body burden. The cadmium concentrations in the total soft tissues far exceeded the EU-limit of 1 μg/g wet weight for cadmium in bivalves. Based on this, selective evisceration of the cadmium-rich kidney and digestive gland during processing can be regarded as a reliable measure to be taken in order to reduce the cadmium content of scallops used for human consumption.

Metal residues, histopathology and presence of parasites in the liver and gills of fourhorn sculpin ( Myoxocephalus quadricornis ) and shorthorn sculpin ( Myoxocephalus scorpius ) near a former lead-zinc mine in East Greenland

ABSTRACT Fourhorn sculpins (Myoxocephalus quadricornis) and shorthorn sculpins (Myoxocephalus scorpius) have been considered suitable local bioindicators for environmental monitoring studies in the Arctic. Because these species share many characteristics, data from the two species have previously been pooled when assessing marine metal contamination. A chemical and histological study was conducted on fourhorn and shorthorn sculpins collected around a contaminated lead‐zinc mine at East Greenland to investigate whether there were any differences in the residues of metals, histopathology and parasites in liver and gills between the two sculpin species. The results demonstrated that concentrations of copper (Cu), zinc (Zn), mercury (Hg) and lead (Pb) were significantly higher in the fourhorn sculpins (p<0.001) while there were no significant differences for arsenic (As) or cadmium (Cd). Furthermore, density of blood vessel fibrosis (p=0.028), prevalence and density of chondroplasia (p=0.002 and p=0.005, respectively), number of mucin‐containing mucous cells (p<0.001) and chloride cells (p<0.001) and mean intensity of colonial Peritricha (p<0.001) were significantly higher in fourhorn sculpin. Based on these results we suggest that pooling the two species when conducting environmental assessments is not recommended as it can lead to incorrect conclusions. We propose that a larger study investigating the biological effects of zinc‐lead mining in Greenland is needed. HighlightsFourhorn sculpins (Myoxocephalus quadricornis) more sensitive to pollution than shorthorn sculpins (Myoxocephalus scorpius).Metal residues, histological changes and presence of parasites were species‐specific.Different sculpin species should not be pooled together as pollution biomarkers.

Applying Chemometrics to Determine Dispersion of Mine Tailing-Affected Sediments from Submarine Tailing Disposal in Bøkfjorden, Northern Norway

Mine tailing management is one of the largest environmental issues related to mining operation. This study uses chemometrics to assess the dispersion of iron mine tailing-affected sediments in Bøkfjorden, Northern Norway. Metal concentrations (Al, As, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Zn) and physico-chemical sediment characteristics (conductivity, organic matter, sulphate, chloride, grain size, CaCO3, pH) were analysed in seven sediment cores collected in a transect out of the fjord along with two reference cores. Results of hierarchical cluster analysis and principal component analysis allowed to distinguish between mine tailing-affected and non-affected sediments. Non-affected sediments were especially characterised by high levels of organic matter whilst mine tailing-affected sediments varied significantly in sediment characteristics depending on location in the fjord. Crucial parameters to reveal mine tailing-affected sediments varied between the target metal Fe along with metals of Cd and Mn, albeit less significant. Variations in mine tailing-affected sediment characteristics could be attributed to other anthropogenic activities in the fjord. Despite potential disturbances, chemometrics made it possible to identify dispersion of mine tailing-affected sediments to cover the inner and middle parts of the fjord. The study demonstrates the advantage of applying chemometrics on complex fjord systems, which in this case was used to distinguish mine tailing-affected sediments from areas with elevated levels of metals not necessarily related to the mine.

Mytilus trossulus in NW Greenland is genetically more similar to North Pacific than NW Atlantic populations of the species

Changes in climate-related factors such as ice coverage, water temperature, and ocean currents have been proposed to facilitate an increased interchange of species in the High Arctic between the Pacific and Atlantic oceans. In Greenland, the colonization of the mussel species Mytilus edulis has been suggested to have occurred recently and exclusively from Northwest Atlantic populations. The source population for its sibling species Mytilus trossulus is however unknown, and therefore we aimed to explore its genetic origin. Using 54 SNP markers, M. trossulus was identified from three Greenland blue mussel populations collected in 2012–2014 and the relative similarities to Northwest Atlantic and North Pacific M. trossulus populations were assessed. Populations were found to fall into two clades. The North Pacific is the most likely source for the northern Greenland M. trossulus probably as a result of occasional postglacial long-distance dispersal through the Bering Strait.Our findings in M. trossulus are in agreements with the predictions that climate change will, in addition to driving a northward expansion of temperate-boreal species into the Arctic Ocean, increase the rate of trans-Arctic interchange between the Atlantic and the Pacific oceans.

Comparison of heavy metals, parasites and histopathology in sculpins (Myoxocephalus spp.) from two sites at a lead-zinc mine in North East Greenland

Abstract The former lead‐zinc mine at Mestersvig, Greenland, continues to contaminate the surrounding environment despite its operations ceasing over 50 years ago. Elevated concentrations of heavy metals are found in water, sediment and biota in the terrestrial, freshwater and marine environments. To shed light on the present contamination and its potential effects on local fish we investigated gill and liver histology of sculpins (Myoxocephalus spp.) around the former mining area. Two species of sculpins were caught; shorthorn sculpins (M. scorpius; n = 16) and fourhorn sculpins (M. quadricornis; n = 17) at a contaminated site, Nyhavn, and shorthorn sculpins (M. scorpius; n = 25) at the reference site. In a previous study we found histopathological changes in the tissues of the sculpins, and we suspected this to be related to elevated heavy metal tissue concentrations. Concentrations of Fe, Hg, Mn, Pb, Se and Zn were significantly higher in sculpins at Nyhavn compared to the reference site. Reference NOED and LOEC thresholds for biochemistry, tissue lesions, growth, survival and reproduction for hepatic Hg, As, Cd and Pb from the ERED database were exceeded in both sculpin species. Histopathological investigations of the sculpins gills revealed significant increases in the prevalence of hyperplastic epithelium, inflammation, intensity of neutral and total mucus cells and chloride cells along with an increased infection of colonial Peritricha. At the contaminated Nyhavn site, fourhorn sculpins had a significantly higher prevalence of chondroplastic tissue and intensity of neutral, mixed and total mucus cells in the gills compared to the shorthorn sculpins. The data indicate that both sculpin species could be useful indicator species for environmental monitoring of metal pollution in Arctic areas. However, confounding effects of gender and species should be investigated further. Effects on other biomarkers as well as baseline measurements should be included in future environmental monitoring efforts around mining activities in Greenland. HighlightsMestersvig mine Pb‐Zn mine in East Greenland has led to significant metal pollution.Gill and liver morphology of fourhorn and shorthorn sculpins was evaluated as bio‐indicators.Significant pathologies were found and were related to metal concentrations.Sculpin pathology may be suitable health indicators mining activity effects.These should be enrolled in EIAs of raw material industrialisation in the Arctic.