Effects of exposure to environmentally relevant concentrations of lead (Pb) on expression of stress and immune-related genes, and microRNAs in shorthorn sculpins (Myoxocephalus scorpius).

Old lead-zinc (Pb-Zn) mining sites in Greenland have increased the environmental concentration of Pb in local marine organisms, including the shorthorn sculpin. Organ metal concentrations and histopathology have been used in environmental monitoring programs to evaluate metal exposure and subsequent effects in shorthorn sculpins. So far, no study has reported the impact of heavy metals on gene expression involved in metal-related stress and immune responses in sculpins. The aim of this study was to investigate the effect of exposure to environmentally relevant waterborne Pb (0.73 ± 0.35 µg/L) on hepatic gene expression of metallothionein (mt), immunoglobulin M (igm), and microRNAs (miRNAs; mir132 and mir155) associated with immune responses in the shorthorn sculpin compared to a control group. The mt and igm expression were upregulated in the Pb-exposed group compared to the control group. The transcripts of mir132 and mir155 were not different in sculpins between the Pb-exposed and control group; however, miRNA levels were significantly correlated with Pb liver concentrations. Furthermore, there was a positive correlation between liver Pb concentrations and igm, and a positive relationship between igm and mir155. The results indicate that exposure to Pb similar to those concentrations reported in in marine waters around Greenland Pb-Zn mine sites influences the mt and immune responses in shorthorn sculpins. This is the first study to identify candidate molecular markers in the shorthorn sculpins exposed to waterborne environmentally relevant Pb suggesting mt and igm as potential molecular markers of exposure to be applied in future assessments of the marine environment near Arctic mining sites.

Element concentrations, histology and serum biochemistry of arctic char (Salvelinus alpinus) and shorthorn sculpins (Myoxocephalus scorpius) in northwest Greenland.

The increasing exploratory efforts in the Greenland mineral industry, and in particular, the proposed rare earth element (REE) mining projects, requires an urgent need to generate data on baseline REE concentrations and their potential environmental impacts. Herein, we have investigated REE concentrations in anadromous Arctic char (Salvelinus alpinus) and shorthorn sculpins (Myoxocephalus scorpius) from uncontaminated sites in Northwest Greenland, along with the relationships between the element concentrations in gills and liver, and gill histology and serum biochemical parameters. Concentrations of arsenic, silver, cadmium, cerium, chromium, copper, dysprosium, mercury, lanthanum, neodymium, lead, selenium, yttrium, and zinc in gills, liver and muscle are presented. No significant statistical correlations were observed between element concentrations in different organs and gill histology or serum biochemical parameters. However, we observed positive relationships between age and histopathology, emphasizing the importance of including age as a co-variable in histological studies of fish. Despite no element-induced effects were observed, this study is considered an important baseline study, which can be used as a reference for the assessment of impacts of potential future REE mine sites in Greenland.

Histopathological effects of short-term aqueous exposure to environmentally relevant concentration of lead (Pb) in shorthorn sculpin (Myoxocephalus scorpius) under laboratory conditions.

Shorthorn sculpin (Myoxocephalus scorpius) has been used as a sentinel species for environmental monitoring, including heavy metal contamination from mining activities. Former lead-zinc (Pb-Zn) mines in Greenland resulted in elevated concentrations of metals, especially Pb, in marine biota. However, the potential accumulation of Pb and effects of the presence of Pb residues in fish on health of sculpins observed in the field have not been validated in laboratory experiments. Therefore, our aim was to validate field observation of shorthorn sculpin via controlled laboratory exposure to environmentally relevant concentrations of dissolved Pb. We evaluated the effects of a short-term (28 days) exposure to Pb on Pb residues in sculpin blood, gills, liver, and muscle and the morphology of gills and liver. The highest level of Pb was found in the gills, followed by muscle and then liver. Pb levels in liver, gills, and blood of Pb-exposed sculpins were significantly higher than those in control fish, showing that blood is suitable for assessing Pb accumulation and exposure in sculpins. Histopathological investigations showed that the severity score of liver necrosis and gill telangiectasia of Pb-exposed sculpins was significantly greater than in control fish. The number of mucous cells in gills was positively correlated with Pb concentrations in organs. Overall, the results validated field observation for the effects of Pb on wild sculpin and contributed to the improved use of the shorthorn sculpin as sentinel species for monitoring contamination from Pb mines in the Arctic.

Bioaccumulation of rare earth elements in juvenile arctic char (Salvelinus alpinus) under field experimental conditions.

Few ecotoxicological studies exist on the accumulation and effects of rare earth elements (REEs) in fish, particularly on Arctic species. In southwest Greenland, there are currently several advanced exploration REE mining projects. The aim of this study was to investigate accumulation of REEs in native fish species. Juvenile arctic chars, Salvelinus alpinus, were pulse-exposed to cerium (Ce), lanthanum (La) and yttrium (Y) using an in-situ flow-through system over a period of 15 days. Results showed that the arctic char accumulated most REEs in the gills > liver > muscle. We also demonstrated the ability of the arctic char to rapidly excrete the REEs throughout the experiment, where levels of post exposure accumulation also declined throughout the period. These results demonstrate the importance of further studies on accumulation of REE in the arctic char native to the site of future mining operations. Long-term exposure will most likely result in accumulation of REEs in arctic char, and the effects and accumulation patterns of this should be explored further.

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

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.

Diffusion or advection? Mass transfer and complex boundary layer landscapes of the brown alga Fucus vesiculosus.

The role of hyaline hairs on the thallus of brown algae in the genus Fucus is long debated and several functions have been proposed. We used a novel motorized set-up for two-dimensional and three-dimensional mapping with O2 microsensors to investigate the spatial heterogeneity of the diffusive boundary layer (DBL) and O2 flux around single and multiple tufts of hyaline hairs on the thallus of Fucus vesiculosus. Flow was a major determinant of DBL thickness, where higher flow decreased DBL thickness and increased O2 flux between the algal thallus and the surrounding seawater. However, the topography of the DBL varied and did not directly follow the contour of the underlying thallus. Areas around single tufts of hyaline hairs exhibited a more complex mass-transfer boundary layer, showing both increased and decreased thickness when compared with areas over smooth thallus surfaces. Over thallus areas with several hyaline hair tufts, the overall effect was an apparent increase in the boundary layer thickness. We also found indications for advective O2 transport driven by pressure gradients or vortex shedding downstream from dense tufts of hyaline hairs that could alleviate local mass-transfer resistances. Mass-transfer dynamics around hyaline hair tufts are thus more complex than hitherto assumed and may have important implications for algal physiology and plant-microbe interactions.

Ecotoxicological investigation of the effect of accumulation of PAH and possible impact of dispersant in resting high arctic copepod Calanus hyperboreus.

Due to high lipid content and a slow metabolism, there is a higher risk of bioaccumulation of oil compounds in Arctic than in temperate copepods. There is also a concern that the bioavailability of oil compounds is higher when oil is dispersed with dispersants. The purpose of this project was to increase the knowledge on how the use of dispersants on an oil spill may affect the passive uptake of PAHs in resting high arctic copepods using Calanus hyperboreus as a model organism. To evaluate this, resting high arctic C. hyperboreus were caught in Disko Bay at>250 meters depth, November 2013, and subsequent experimental work was initiated immediately after, at nearby Arctic Station at Disko Island Western Greenland. C. hyperboreus females were incubated in phenanthrene (111, 50 and 10 nM), pyrene (57, 28 and 6 nM) and benzo(a) pyrene (10, 5 and 1 nM) for three days in treatments with and without oil (corn oil) and dispersant (AGMA DR372). After exposure, the highest measured concentrations of respectively phenanthrene, pyrene and benzo(a) pyrene in the copepods were 129, 30 and 6 nmol PAH g female(-1). Results showed that with addition of oil and dispersant to the water, the accumulation of PAH was significantly reduced, due to the deposition of the PAHs in the oil phase, decreasing the available PAHs for copepod uptake. While PAH metabolites and a depuration of the PAHs were observed, the copepods still contained PAHs after 77 days of incubation in clean seawater. Differences of treatments with and without oil and dispersant on the egg production were not statistically conclusive, although it is the most likely an effect of the highly variable day-to-day egg production between individual copepods. Equally, although there was an indication that the addition of dispersant and oil increased the mortality rate, there was no statistical difference.

Evaluating pyrene toxicity on Arctic key copepod species Calanus hyperboreus.

Calanus hyperboreus is a key species in the Arctic regions because of its abundance and role in the Arctic food web. Exploitation of the off shore oil reserves along Western Greenland is expected in the near future, and it is important to evaluate the acute and chronic effects of oil emissions to the ecosystem. In this study C. hyperboreus females were exposed to concentrations of 0, 0.1, 1, 10 and 100 nM pyrene and saturated concentrations measured to ~300 nM. Daily quantification of egg and faecal pellet production showed significant decreases in the pellet production, while the egg production was unaffected. The hatching success was also unaffected, although the total reproductive output was reduced with increased pyrene concentrations. Accumulation of pyrene in the copepods was higher in feeding than starving females and only trace amounts of the phase I metabolite 1-hydroxypyrene, were found. Lowered reproductive output, reduced grazing, and reduced ability to metabolize pyrene suggest that oil contamination may constitute a risk to C. hyperboreus recruitment, energy transfer in the food web and transfer of pyrene to higher trophic levels.