Andrew Atfield

Reduced salinities compromise the thermal tolerance of hypersaline specialist diving beetles.

Inland saline waters are globally threatened habitats that harbour unique assemblages of specialist invertebrates. In many Mediterranean regions, irrigation associated with intensive agriculture is lowering the salinity of these habitats, resulting in the loss of their specialist biota, although the mechanisms by which reductions in salinity lead to species loss are poorly understood. In the present study, the effects of reduced salinity on the temperature tolerance and thermal acclimatory abilities of two related species of hypersaline water beetles, Nebrioporus baeticus (Schaum) and Nebrioporus ceresyi (Aubé), are explored. Both upper (UTL) and lower thermal limits (LTL) are assessed, and both salinity and temperature are found to influence the thermal biology of Nebrioporus. Mean UTLs are greater in individuals of both species acclimated at high salinities, with salinity appearing to be more important than acclimation temperature in determining UTL. In both taxa, the lowest mean LTLs are recorded in individuals acclimated at the highest salinities and lowest temperatures; temperature‐dependent acclimation is only reported after exposure to relatively high salinities. The data show that salinity influences the thermal tolerance and acclimatory ability of these hypersaline beetles, and that lowered salinity compromises the ability of adult Nebrioporus to cope with both heat and cold. Such an effect may partly explain why specialist species are lost from hypersaline habitats subject to salinity reductions, and suggests that ongoing reduction in salinity may compromise the ability of such specialist taxa to cope with rapid climate change.

Use of an exposure chamber to maintain aqueous phase nanoparticle dispersions for improved toxicity testing in fish

A novel chamber for maintaining aqueous phase dispersions of nanoparticles (NPs) to enable improved toxicity testing in larval zebrafish was developed. Aqueous concentrations were within 80% of initial NP concentrations, and the 96-h median lethal concentration (LC50) values were highly reproducible (coefficient of variation <0.16, n = 3 tests). Significantly lower toxicity for each NP tested (Ag, Cu, and TiO2 NPs) in static beakers suggested that traditional acute toxicity tests may underestimate aqueous phase toxicity of NPs.

Developmental toxicity of metaldehyde in the embryos of Lymnaea stagnalis (Gastropoda: Pulmonata) co-exposed to the synergist piperonyl butoxide.

Metaldehyde is a widely used molluscicide in countries where damage to crops from slugs and snails is a major problem associated with warm and wet winters. In the UK it is estimated that over 8% of the area covered by arable crops is treated with formulated granular bait pellets containing metaldehyde as the main active ingredient. Metaldehyde is hydrophilic (log Kow=0.12), water soluble (200 mg·L(-1) at 17 °C) and has been detected in UK surface waters in the concentration range of typically 0.2-0.6 μg·L(-1) (maximum 2.7 μg·L(-1)) during 2008-2011. In the absence of chronic data on potential hazards to non-target freshwater molluscs, a laboratory study was conducted to investigate the impact of metaldehyde on embryo development in the gastropod Lymnaea stagnalis (RENILYS strain) and using zinc as a positive control. L. stagnalis embryos were exposed to metaldehyde under semi-static conditions at 20±1 °C and hatching success and growth (measured as shell height and intraocular distance) examined after 21 d. Exposure concentrations were verified using HPLC and gave 21 d (hatching)NOEC and (hatching)LOEC mean measured values of 36 and 116 mg MET·L(-1), respectively (equal to the 21 d (shell height)NOEC and (shell height)LOEC values). For basic research purposes, a second group of L. stagnalis embryos was co-exposed to metaldehyde and the pesticide synergist piperonyl butoxide (PBO). Co-exposure to the PBO (measured concentrations between 0.47-0.56 mg·L(-1)) reduced hatching success from 100% to 47% and resulted in a 30% reduction in embryo growth (shell height) in snail embryos co-exposed to metaldehyde at 34-36 mg·L(-1) over 21 d. In conclusion, these data suggest mollusc embryos may have some metabolic detoxication capacity for metaldehyde and further work is warranted to explore this aspect in order to support the recent initiative to include molluscs in the OECD test guideline programme.

The comparative biology of diving in two genera of European Dytiscidae (Coleoptera)

Surfacing behaviour is fundamental in the ecology of aquatic air‐breathing organisms; however, it is only in vertebrates that the evolutionary ecology of diving has been well characterized. Here, we explore the diving behaviour of dytiscid beetles, a key group of surface‐exchanging freshwater invertebrates, by comparing the dive responses of 25 taxa (Deronectes and Ilybius spp.) acclimated at two temperatures. The allometric slopes of dive responses in these dytiscids appear similar to those of vertebrate ectotherms, supporting the notion that metabolic mode shapes the evolution of diving performance. In both genera, beetles spend more time submerged than on the surface, and surface time does not vary with the temperature of acclimation. However, presumably in order to meet increased oxygen demand at higher temperatures, Deronectes species increase surfacing frequency, whereas Ilybius species decrease dive time, an example of ‘multiple solutions.’ Finally, widespread northern species appear to possess higher diving performances than their geographically restricted southern relatives, something which may have contributed to their range expansion ability.

Sub-lethal effects of waterborne exposure to copper nanoparticles compared to copper sulphate on the shore crab (Carcinus maenas)

The toxicity of soluble copper (Cu) to marine organisms is reasonably well described. However, the hazard of Cu engineered nanomaterial (ENMs) is poorly understood. The aim of the present study was to compare the toxicity of Cu ENMs to Cu as CuSO4 in the shore crab, Carcinus maenas. The crabs were exposed via the water using a semi-static approach to 0.2 or 1mgL-1 of Cu ENMs or 1mgL-1 of Cu as CuSO4. Gills, hepatopancreas, chela muscle and haemolymph were collected at days 0, 4 and 7 for the body burden of Cu, histology and biochemical analysis [thiobarbituric acid reactive substances (TBARS) and total glutathione (GSH)]. Nominal exposure concentrations of both the ENMs and the metal salt were maintained at over 80% in each treatment throughout the experiment. By day 7, 54% mortality was recorded in the 1mgL-1 CuSO4 treatment, compared to just 21% in the 1mgL-1 Cu ENM-exposed crabs. The target organs for Cu accumulation were similar for both forms of Cu with highest concentrations in the gills, particularly the posterior gills; followed by the hepatopancreas, and with the lowest concentrations in the chela muscle. No changes were observed in the osmolarity of the haemolymph (ANOVA, P>0.05). TBARS were measured as an indicator of lipid peroxidation and showed the greatest change in the anterior and posterior gills and hepatopancreas of animals exposed to 1mgL-1 Cu ENMs (ANOVA or Kruskal-Wallis, P<0.05). No statistically significant changes in total GSH were observed (ANOVA, P>0.05; n=6 crabs per treatment). Histological analysis revealed organ injuries in all treatments. The types of pathologies observed in the Cu ENM treatments were broadly similar to those of the Cu as CuSO4 treatment. Overall, the target organs and Cu accumulation from Cu ENMs were comparable to that following exposure to Cu as CuSO4, and although there were some differences in the sub-lethal effects, the metal salt was more acutely toxic.