Influence of nanoplastic surface charge on eco-corona formation, aggregation and toxicity to freshwater zooplankton.

Concerns about possible environmental implications of nano- and micro-plastics are continuously raising. Hence, comprehensive understanding of their behaviour, bioaccumulation and toxicity potential is required. Nevertheless, systematic studies on their fate and possible effects in freshwaters, as well as the influence of particle-specific and environmental factors on their behaviour and impacts are still missing. The aims of the present study are thus two-fold: (i) to examine the role of the surface charge on nanoplastic stability and acute effects to freshwater zooplankton; (ii) to decipher the influence of the refractory natural organic matter (NOM) on the nanoplastic fate and effects. Amidine and carboxyl-stabilized polystyrene (PS) spheres of 200 nm diameter characterized by opposite primary surface charges and neutral buoyancy were selected as model nanoplastics. The results demonstrated that the surface functionalization of the polystyrene nanoplastics controls their aggregation behaviour. Alginate or Suwannee River humic acid (SRHA) modified significantly the surface charge of positively-charged amidine PS nanoplastic and the aggregation state, while had no significant influence on the negatively-charged carboxyl PS nanoplastic. Both amidine and carboxyl PS nanoplastics were ingested by the zooplankton and concentrated mainly in the gut of water flea Daphnia magna and larvae Thamnocephalus platyurus, and the stomach of rotifer Brachionus calyciflorus. Amidine PS nanoplastic was more toxic than carboxyl one. The toxicity decreased in the order D. magna (48 h -immobilization) > B. calyciflorus (24 h - lethality) > T. platyurus (24 h - lethality). Alginate or SRHA reduced significantly the toxicity of both amidine and carboxyl PS nanoplastics to the studied zooplankton representatives. The implications of this laboratory study findings to natural environment were discussed.

Preparation of Fluorescent Microcystin Derivatives by Direct Arginine Labelling and Their Biological Evaluation.

Microcystin is the most prevalent toxin produced by cyanobacteria and poses a severe threat to livestock, humans and entire ecosystems. We report the preparation of a series of fluorescent microcystin derivatives by direct arginine labelling of the unprotected peptides at the guanidinium side chain. This new method allows a simple late-stage diversification strategy for native peptides devoid of protecting groups under mild conditions. A series of fluorophores were conjugated to microcystin-LR in good to very good yield. The fluorescent probes displayed biological activity comparable to that of unlabelled microcystin, in both phosphatase inhibition assays and toxicity tests on the crustacean Thamnocephalus platyurus. In addition, we demonstrate that the fluorescent probes penetrated Huh7 cells. Whole-animal imaging was performed on T. platyurus: labelled compound was mainly observed in the digestive tract.

Changes in agglomeration of fullerenes during ingestion and excretion in Thamnocephalus platyurus.

The crustacean Thamnocephalus platyurus was exposed to aqueous suspensions of fullerenes C(60) and C(70) . Aqueous fullerene suspensions were formed by stirring C(60) and C(70) as received from a commercial vendor in deionized water (termed aqu/C(60) and aqu/C(70) ) for approximately 100 d. The Z-average (mean hydrodynamic) diameters of aqu/C(60) and aqu/C(70) aggregates as measured by dynamic light scattering were 517 ± 21 nm and 656 ± 39 nm (mean ± 95% confidence limit), respectively. Exposure of T. platyurus to fullerene suspensions resulted in the formation of dark masses in the digestive track visible under a stereo microscope (×40 magnification). Fullerene ingestion over 1 h of exposure was quantitatively determined after extraction and analysis by high-performance liquid chromatography-mass spectrometry (HPLC-MS). One-hour exposures (at 3 mg/L and 6 mg/L) resulted in aqu/C(60) burdens of 2.7 ± 0.4 µg/mg and 6.8 ± 1.5 µg/mg wet weight, respectively. Thin-section transmission electron microscopy (TEM) images of aqu/C(60) -exposed T. platyurus showed the formation in the gut of fullerene agglomerates (5-10 µm) that were an order of magnitude larger than the suspended fullerene agglomerates. Upon excretion, the observed fullerene agglomerates were in the 10- to 70-µm size range and settled to the bottom of the incubation wells. In contrast to the control polystyrene microspheres, which dispersed after depuration, the aqu/C(60) agglomerates (greater than two orders of magnitude larger than the suspended fullerenes) remained agglomerated for up to six months. When exposed to fullerenes, T. platyurus shows the potential to influence agglomerate size and may facilitate movement of these nanoparticles from the water column into sediment.

Early Distal-less expression in a developing crustacean limb bud becomes restricted to setal-forming cells.

Distal-less (Dll) plays a well-known role in patterning the distal limb in arthropods. However, in some taxa, its expression even during early limb development is not always limited to the distal limb. Here, I trace the expression of Distal-less in a crustacean (Thamnocephalus platyurus) from the early limb bud to later stages of limb development, a period that includes differentiation of juvenile and adult morphology. During early development, I find two distinct types of DLL expression: one correlated with proximal distal leg patterning and the other restricted to setal-forming cells. Later in development, all the DLL expression is restricted to setal-forming cells. Based on the particular cells expressing DLL, I hypothesize an ancestral role for Dll function in the formation accessory cells of sensilla.

Comparisons of stress proteins and soluble carbohydrate in encysted embryos of Artemia franciscana and two species of Parartemia.

We compared stress proteins (p26, artemin, hsp70) and alcohol-soluble carbohydrates (ASC) in cysts of Artemia franciscana and two as yet un-named species populations of Parartemia, the brine shrimp endemic to Australia. The small stress proteins and molecular chaperones, p26 and artemin, previously thought to be restricted to Artemia, and present in very large amounts in its encysted embryos (cysts), were also detected by western blotting in Parartemia cysts, even though roughly 85-100 million years have passed since these genera diverged. We interpret this finding as further evidence for the adaptive importance of these proteins in coping with the severe stresses these encysted embryos endure. As expected, hsp70 was present in all three groups of cysts, but apparently at somewhat lower concentrations in those of Parartemia. Based on measurements of ASC we propose that the disaccharide trehalose, critical for desiccation tolerance in many animal cells, has probably also been maintained in the metabolic repertoire of Parartemia whose cysts have well developed tolerance to severe desiccation.

Sensitivity of ecotoxicological tests in analysis of superfund sites.

During the analysis of environmental risk posed by hazardous waste disposal sites, ecological impact on whole ecosystems should be assessed. It requires a complex testing scheme where surrogate species represent key elements of the ecosystem. However, different organisms are exposed to a differing degree, also, their sensitivity to the same contaminant may vary. A possible way to determine which test reflects most the actual toxic conditions, correlation can be calculated between the measured ecological parameter (such as growth inhibition, mortality, etc.) and a contaminant gradient. The basic aim of this study was to determine which ecotoxicological test shows the best correlation with the measured analytical parameters. The selected tests were Lemna minor (representing primary producers), Thamnocephalus platyurus (a primary consumer organism) and Vibrio fischeri (decomposer). When testing soil samples, the Thamnocephalus test showed excellent consistency with most contaminants but was oversensitive in the case of groundwater samples. The Vibrio fischeri bioluminescence inhibition test (ToxAlert) behaved in a different way, reflecting well the distribution of most contaminants in groundwater samples. Finally, Lemna test proved to be completely inadequate.