Cytotoxicity and Physiological Effects of Silver Nanoparticles on Marine Invertebrates.

Silver nanoparticles (AgNPs) incorporation in commercial products is increasing due to their remarkable physical and chemical properties and their low cost on the market. Silver has been known for a long time to be highly toxic to bacterial communities, aquatic organisms, and particularly to marine biota. Strong chloro-complexes dominate Ag speciation in seawater and facilitate its persistence in dissolved form. It has a great impact on marine organisms because low concentration of silver can lead to strong bioaccumulation, partly because the neutral silver chloro complex (AgCl0) is highly bioavailable. Owing to the fact that estuaries and coastal areas are considered as the ultimate fate for AgNPs, the study of their toxic effects on marine invertebrates can reveal some environmental risks related to nanosilver exposure. In an attempt to reach this goal, many invertebrate taxa including mollusks, crustaceans, echinoderms and polychaetes have been used as biological models. The main findings related to AgNP toxicity and marine invertebrates are summarized hereafter. Some cellular mechanisms involving nano-internalization (cellular uptake, distribution and elimination), DNA damaging, antioxidant cellular defenses and protein expression are discussed. Physiological effects on early stage development, silver metabolic speciation, immune response, tissue damaging, anti-oxidant effects and nano-depuration are also described. Finally, we paid attention to some recent interesting findings using sea urchin developmental stages and their cells as models for nanotoxicity investigation. Cellular and physiological processes characterizing sea urchin development revealed new and multiple toxicity mechanisms of both soluble and nano forms of silver.

Cell-mediated immune response of postmetamorphic sea urchin juveniles against infectious stages of diatom Cylindrotheca closterium (Bacillariophyceae).

In this study, successive infectious stages by diatom Cylindrotheca closterium (Bacillariophyceae) are described for the first time during the early development of sea urchin at low temperature (8°C). Diatom cell-types enclosed or not by typical theca were capable of infection. As an immune response, red spherulocytes and amoebocytes migrated towards infested areas and restrained the infection spreading over shells in 2- and 3-month old urchins. Only amoebocyte cells appeared to be involved in the immune reaction of 1-month old specimens which turned out to be a less effective fence to stop infestation. The effectiveness of the inflammatory process caused by diatoms within sea urchin spines seemed to vary as a function of urchin age, cohort's type and individual cell pool.

Silver nanoparticles and dissolved silver activate contrasting immune responses and stress-induced heat shock protein expression in sea urchin.

Using immune cells of sea urchin Strongylocentrotus droebachiensis in early development as a model, the cellular protective mechanisms against ionic and poly(allylamine)-coated silver nanoparticle (AgNPs; 14 ± 6 nm) treatments at 100 µg L-1 were investigated. Oxidative stress, heat shock protein expression, and pigment production by spherulocytes were determined as well as AgNP translocation pathways and their multiple effects on circulating coelomocytes. Sea urchins showed an increasing resilience to Ag over time because ionic Ag is accumulated in a steady way, although nanoAg levels dropped between 48 h and 96 h. A clotting reaction emerged on tissues injured by dissolved Ag (present as chloro-complexes in seawater) between 12 h and 48 h. Silver contamination and nutritional state influenced the production of reactive oxygen species. After passing through coelomic sinuses and gut, AgNPs were found in coelomocytes. Inside blood vessels, apoptosis-like processes appeared in coelomocytes highly contaminated by poly(allylamine)-coated AgNPs. Increasing levels of Ag accumulated by urchins once exposed to AgNPs pointed to a Trojan-horse mechanism operating over 12-d exposure. However, under short-term treatments, physical interactions of poly(allylamine)-coated AgNPs with cell structures might be, at some point, predominant and responsible for the highest levels of stress-related proteins detected. The present study is the first report detailing nano-translocation in a marine organism and multiple mechanisms by which sea urchin cells can deal with toxic AgNPs. Environ Toxicol Chem 2017;36:1872-1886. © 2016 SETAC.

Physiological effects and cellular responses of metamorphic larvae and juveniles of sea urchin exposed to ionic and nanoparticulate silver.

The widespread use of silver nanoparticles (AgNPs) would likely result in their discharge into wastewater and inevitable release in densely populated coastal areas. It is known that AgNPs can cause harmful effects to marine fauna, but how they affect development stages is still an open question. In order to understand in details how polymer-coated AgNPs (PAAm-AgNPs) (from 0.19 to 4.64mM as Ag) can affect critical stages of marine invertebrate development, metamorphic larvae and juveniles of sea urchins were used as biological models. Multidimensional scaling (MDS) approach based on Bray-Curtis similarity matrix with PERMANOVA showed organisms in a multivariate space undergoing through different physiological conditions as a function of time, chemical forms of silver, nominal concentrations, and presence or absence of food. Sublethal effects such as lethargy, oedema and immobility mainly characterized PAAm-AgNPs effects with juveniles and postlarvae, whereas necrosis and death arose in Ag(+) conditions in short-term tests. Chronically exposed metamorphic larvae had their morphogenic processes interrupted by PAAm-AgNPs and a high mortality rate was observed in recovery period. On the contrary, Ag(+) ions caused progressive mortality during exposure, but a quick recovery in uncontaminated seawater was observed. By means of fluorescent markers we showed that nanosilver could be transferred between consecutive stages (swimming larvae and postlarvae) and highlighted how important is food to enhance PAAm-AgNPs uptake. Using TEM we observed that unfed juveniles had nanosilver aggregates mostly restricted to their coelomic sinuses, while metamorphic larvae already had nano-contamination overspread in different tissues and blastocoel. Our main hypothesis for nanotoxicity of PAAM-AgNPs relies on the slow dissolution of nano-core over time, but in this study the effects of particulate silver form itself are also evoked. Main mechanisms governing tissular and cellular responses to nano-intoxication such as inflammatory response and detoxification based on the role of sentinel cells (peritoneal cells and coelomocytes) for general homeostasis are discussed. This paper is first to detail physiological states, main uptake routes and cellular response against polymer-coated AgNPs in developmental stages of marine invertebrate species.

Toxicity mechanisms of ionic silver and polymer-coated silver nanoparticles with interactions of functionalized carbon nanotubes on early development stages of sea urchin.

Exposures of aquatic organisms to multiple contaminants are likely to take place in estuarine and coastal areas and combined effects on early life stages have to be examined. Among emerging contaminants, ionic silver (Ag(+)) and silver nanoparticles (AgNps) have demonstrated contrasting effects on marine invertebrates, but their interactions with functionalized carbon nanotubes (f-SWCNTs) have not yet been investigated in details. In order to observe the impacts and understand the toxicity mechanism of Ag(+) and polymer-coated AgNps, and their combined effects with f-SWCNTs, successive development stages of embryos of sea urchin, Strongylocentrotus droebachiensis, were exposed to Ag(+), AgNps and f-SWCNTs, separately and in mixtures using moderate environmental concentrations. We also assessed long-term effects of treatments under recovery conditions. Morphological endpoints such as archenteron elongation, primary and secondary mesenchyme cells fate, pigment cells migration, spiculogenic cells and gut development indicated different effects of silver and nanosilver forms during successive development stages. Whereas Ag(+) induced vegetalization and extrusion of mesenchyme cells on early embryos; f-SWCNTs+Ag(+) strongly interfered with gut regionalization in late larvae. Sensitive blastocoelar cells got vacuolized and shapeless with AgNps, but not with mixtures with f-SWCNTs. Increased concentrations of Ag(+) and f-SWCNTs+Ag(+) led to the most disruptive effects during development, but f-SWCNTs+Ag(+) caused the highest mortality rate during the recovery period, which indicated far-reaching effects driven by f-SWCNTs and their ability to keep silver more available during exposure period.

Sistemática de Rhabdepyris Kieffer (Hymenoptera, Bethylidae) Amazônicos com antenas pectinadas

Rhabdepyris opistolatus sp. nov., R. longimerus sp. nov., R. pleurorrectus sp. nov., R. latimerus sp. nov. and R. pectinatus sp. nov. with ramose antennae from Brazilian Amazon rainforest are described and illustrated.

São descritas e ilustradas Rhabdepyris opistolatus sp. nov., R. longimerus sp. nov., R. pleurorrectus sp. nov., R. latimerus sp. nov. e R. pectinatus sp. nov. com antenas pectinadas da floresta amazônica brasileira.