To compare aquatic organisms’ responses to the toxicity of copper oxide (CuO) nanoparticles (NPs) with those of CuO microparticles (MPs) and copper (Cu) ions, a global metabolomics approach was employed to investigate the changes of both polar and nonpolar metabolites in microalga Chlorella vulgaris after 5-day exposure to CuO NPs and MPs (1 and 10 mg/L), as well as the corresponding dissolved Cu ions (0.08 and 0.8 mg/L). Unchanged growth, slight reactive oxygen species production, and significant membrane damage (at 10 mg/L CuO particles) in C. vulgaris were demonstrated. A total of 75 differentiated metabolites were identified. Most metabolic pathways perturbed after CuO NPs exposure were shared by those after CuO MPs and Cu ions exposure, including accumulation of chlorophyll intermediates (max. 2.4–5.2 fold), membrane lipids remodeling for membrane protection (decrease of phosphatidylethanolamines (min. 0.6 fold) and phosphatidylcholines (min. 0.2–0.7 fold), as well as increase of phosphatidic acids (max. 1.5–2.9 fold), phosphatidylglycerols (max. 2.2–2.3 fold), monogalactosyldiacylglycerols (max. 1.2–1.4 fold), digalactosylmonoacylglycerols (max. 1.9–3.8 fold), diacylglycerols (max. 1.4 fold), lysophospholipids (max. 1.8–3.0 fold), and fatty acids (max. 3.0–6.2 fold)), perturbation of glutathione metabolism induced by oxidative stress, and accumulation of osmoregulants (max. 1.3–2.6 fold) to counteract osmotic stress. The only difference between metabolic responses to particles and those to ions was the accumulation of fatty acids oxidation products: particles caused higher fold changes (particles/ions ratio 1.9–3.0) at 1 mg/L and lower fold changes (particles/ions ratio 0.4–0.7) at 10 mg/L compared with ions. Compared with microparticles, there was no nanoparticle-specific pathway perturbed. These results confirm the predominant role of dissolved Cu ions on the toxicity of CuO NPs and MPs, and also reveal particle-specific toxicity from a metabolomics perspective.

The effects of salinity on metal toxicity are complex: not only affecting metal bioaccumulation, but also altering the physiology and sensitivity of organisms. In this study, we used a toxicokinetic-toxicodynamic (TK-TD) model to separate and quantify the dual effects of salinity on copper (Cu) toxicity in a euryhaline clam Potamocorbula laevis. The toxicokinetics of Cu was determined using the stable isotope 65Cu as a tracer at concentrations (10–500 μg L−1) realistic to contaminated environments and at salinities ranging from 5 to 30. At low Cu concentrations (ca. 10 μg L−1), Cu bioaccumulation decreased monotonically with salinity, and the uptake rate constant (ku, 0.546 L g−1 h−1 to 0.213 L g−1 h−1) fitted well with an empirical equation, ku = 1/(1.35 + 0.116·Salinity), by treating salinity as a pseudo-competitor. The median lethal concentrations (LC50s) of Cu were 269, 224, and 192 μg L−1 at salinity 5, 15, and 30, respectively. At high Cu concentrations (ca. 500 μg L−1), elevating salinity were much less effective in decreasing Cu bioaccumulation; whereas Cu toxicity increased with salinity. The increased toxicity could be explained by the increases in Cu killing rates (kks), which were estimated to be 0.44–2.08 mg μg−1 h−1 and were presumably due to the osmotic stress caused by the deviation from the optimal salinity of the clams. The other toxicodynamic parameter, internal threshold concentration (CIT), ranged from 79 to 133 μg−1 g−1 and showed no clear trend with salinity.

Across the planet, winter de-icing practices have caused secondary salinization of freshwater habitats. Many amphibians are vulnerable because of permeable skin and reliance on small ponds, where salinity can be high. Early developmental stages of amphibians are especially sensitive to salt, and larvae developing in salt-polluted environments must osmoregulate through ion exchange in gills. Though ionoregulation in amphibian gills is generally understood, the role of gill morphology remains poorly described. Yet gill structure should affect ionoregulatory capacity, for instance in terms of available surface area. As larval amphibian gills also play critical roles in gas exchange and foraging, changes in gill morphology from salt pollution potentially affect not only osmoregulation, but also respiration and feeding. Here, we used an exposure experiment to quantify salinity effects on larval gill morphology in wood frogs (Rana sylvatica). We measured a suite of morphological traits on gill tufts—where ionoregulation and gas exchange occur—and on gill filters used in feeding. Larvae raised in elevated salinity developed larger gill tufts but with lower surface area to volume ratio. Epithelial cells on these tufts were less circular but occurred at higher densities. Gill filters showed increased spacing, likely reducing feeding efficiency. Many morphological gill traits responded quadratically, suggesting that salinity might induce plasticity in gills at intermediate concentrations until energetic demands exceed plasticity. Together, these changes likely diminish ionoregulatory and respiratory functionality of gill tufts, and compromise feeding functionality of gill filters. Thus, a singular change in aquatic environment from a widespread pollutant appears to generate a suite of consequences via changes in gill morphology. Critically, these changes in traits likely compound the severity of fitness impacts in populations dwelling in salinized environments, whereby ionoregulatory energetic demands should increase respiratory and foraging demands, but in individuals who possess structures poorly adapted for these functions.

Application of bioash from biofuel combustion to soil supports nutrient recycling, but may have unwanted and detrimental ecotoxicological side-effects, as the ash is a complex mixture of compounds that could affect soil invertebrates directly or through changes in their food or habitat conditions. To examine this, we performed laboratory toxicity studies of the effects of wood-ash added to an agricultural soil and the organic horizon of a coniferous plantation soil with the detrivore soil collembolans Folsomia candida and Onychiurus yodai, the gamasid predaceous mite Hypoaspis aculeifer, and the enchytraeid worm Enchytraeus crypticus. We used ash concentrations spanning 0–75 g kg⁻¹ soil. As ash increases pH we compared bioash effects with effects of calcium hydroxide, Ca(OH)2, the main liming component of ash. Only high ash concentrations above 15 g kg⁻¹ agricultural soil or 17 t ha⁻¹ had significant effects on the collembolans. The wood ash neither affected H. aculeifer nor E. crypticus. The estimated osmolalities of Ca(OH)2 and the wood ash were similar at the LC50 concentration level. We conclude that short-term chronic effects of wood ash differ among different soil types, and osmotic stress is the likely cause of effects while high pH and heavy metals is of minor importance.

Metal pollution in the Bohai Sea in China has posed a potential risk on marine organisms. In this work, crabs (Portunus trituberculatus) were sampled from four sites, namely a reference (site 3934) and three metal-polluted (sites 6151, 6351, and 3562) sites, located in the Bohai Sea. Metal concentrations in crab gill tissues were measured using inductively coupled plasma mass spectrometry. Cu, Zn, and Cd in crab samples from S3562 presented the highest concentrations. Particularly, Cu concentration exceeded the marine biological quality standard II. Cd contents in crab samples from all metal-polluted sites exceeded the marine biological quality standard I. Nuclear magnetic resonance-based metabolomics indicated metal pollution-induced immune stresses in crab samples from all metal-polluted sites. Metal pollution in S6151 and S6351 disturbed energy metabolism through differential pathways. For crab samples from S3562, the metabolic profile suggested that metal pollution mainly induced osmotic stress.

Phytoplankton cells in estuary waters usually experience drastic changes in chemical and physical environments due to mixing of fresh and seawaters. In order to see their photosynthetic performance in such dynamic waters, we measured the photosynthetic carbon fixation by natural phytoplankton assemblages in the Jiulong River estuary of the South China Sea during April 24–26 and July 24–26 of 2008, and investigated its relationship with environmental changes in the presence or the absence of UV radiation. Phytoplankton biomass (Chl a) decreased sharply from the river-mouth to seawards (17.3–2.1μgL⁻¹), with the dominant species changed from chlorophytes to diatoms. The photosynthetic rate based on Chl a at noon time under PAR-alone increased from 1.9μgC (μg Chl a)⁻¹L⁻¹ in low salinity zone (SSS<10) to 12.4μgC (μg Chl a)⁻¹L⁻¹ in turbidity front (SSS within 10–20), and then decreased to 2.1μgC (μg Chl a)⁻¹L⁻¹ in mixohaline zone (SSS>20); accordingly, the carbon fixation per volume of seawater increased from 12.8 to 149μgCL⁻¹h⁻¹, and decreased to 14.3μgCL⁻¹h⁻¹. Solar UVR caused the inhibition of carbon fixation in surface water of all the investigated zones, by 39% in turbidity area and 7–10% in freshwater or mixohaline zones. In the turbidity zone, higher availability of CO₂ could have enhanced the photosynthetic performance; while osmotic stress might be responsible for the higher sensitivity of phytoplankton assemblages to solar UV radiation.

Copper (Cu) is an essential metal capable to alter many metabolic and physiological processes in animal species, depending on the environmental concentration and salinity. The present study evaluated the effects of Cu exposure on the metabolism of the blue crab Callinectes sapidus under different osmotic situations. Crabs were acclimated at two different salinities conditions (30 and 2). Subsequently, they were exposed to Cu during 96 h at each salinity and under hypo-osmotic shock. Results demonstrated that Cu exposure increased whole-body oxygen consumption. In addition, the activity of LDH decreased while citrate synthase increased in anterior gills from animals submitted to hypo-osmotic shock. This scenario indicates extra stress caused by sudden environmental osmotic changes, as commonly observed in estuarine environments, when combined with copper exposure. Therefore, the activity of LDH and citrate synthase enzymes might be sensitive indicators for aquatic toxicology studies approaching Cu contamination in estuarine environments.

Benzo[a]pyrene (BaP) is a polycyclic aromatic hydrocarbon (PAH) that is well known for its teratogenic, mutagenic and carcinogenic effects. In this study, we applied metabolomics to investigate the tissue-specific metabolic responses of the Pinctada martensii digestive glands and gills after a short-duration exposure to BaP (1 μg/L and 10 μg/L). After 72 h of exposure to BaP, the majority of metabolite changes were related to osmolytes, energy metabolites, and amino acids. BaP (1 μg/L) accelerated energy deterioration and decreased osmotic regulation, while BaP (10 μg/L) disturbed energy metabolism and increased osmotic stress in the digestive glands. Both BaP doses disturbed osmotic regulation and energy metabolism in the gills. BaP also induced neurotoxicity in both tissues. These findings demonstrated that BaP exhibited tissue-specific metabolic responses in P. martensii. The difference in these metabolite responses between the digestive glands and gills might prove to be suitable biomarkers for indicating exposure to specific marine pollutants.

Marine environment in the Laizhou Bay is potentially contaminated by metals from industrial discharges. In this study, metal concentrations in shrimps Crangon affinis indicated that two typical sites (S6283 and S5283) close to Longkou and Zhaoyuan cities along the Laizhou Bay have been contaminated by metals, including Cd, As, Cu, Ni, Co, and Mn. In particular, Cd and As were the main metal contaminants in S6283. In S5283, however, Cu was the most important metal contaminant. The metabolic responses in the shrimps indicated that the metal pollution in S6283 and S5283 induced disturbances in osmotic regulation and energy metabolism and reduced anaerobiosis, lipid metabolism, and muscle movement. However, alteration in the levels of dimethylglycine, dimethylamine, arginine, betaine, and glutamine indicated that the metal pollution in S5283 induced osmotic stress through different pathways compared to that in S6283. In addition, dimethylamine might be the biomarker of Cu in shrimp C. affinis.

Si has a beneficial effect on improving plant tolerance to salt stress. Nevertheless, the mechanisms of Si in mediating the stress responses are still poorly understood. Glycyrrhiza uralensis Fisch. (G. uralensis), a well-known medicinal plant, possesses vast therapeutic potentials. In the present study, a pot experiment was conducted to investigate the long-term effects of Si on growth and physiobiochemical characteristics in 2-year-old G. uralensis subjected to different levels of salinity. Si markedly affected G. uralensis growth in a salt concentration-dependent manner and had no effect on G. uralensis growth under 6 g/kg NaCl. However, it partly reversed the reduction effect induced by 9 g/kg NaCl. In addition, Si significantly increased the contents of soluble sugar and protein but deceased proline content and thus increased water relations; Si markedly increased the activities of SOD, peroxidase, and CAT and further resulted in decreased MDA content and membrane permeability. Moreover, Si altered the levels of phytohormones and their balances. With correlation analysis and principal component analysis (PCA), root biomass had a significant negative correlation with MDA and membrane permeability while a positive correlation with indole-3-acetic acid and GA₃. The PCA partitioned the total variance into three PCs contributing maximum (88.234%) to the total diversity among the salt stress with or without Si due to the study of various traits. In conclusion, Si exerts a beneficial property on salt-induced harmful effects in G. uralensis by relieving osmotic stress, improving water relations, and alleviating oxidative stress; thus, altering the levels and balance of phytohormones results in improved growth of salt-stressed G. uralensis.