Are shallow-water shrimps proxies for hydrothermal-vent shrimps to assess the impact of deep-sea mining?

Polymetallic seafloor massive sulphide deposits are potential targets for deep-sea mining, but high concentrations of metals (including copper - Cu) may be released during exploitation activities, potentially inducing harmful impact. To determine whether shallow-water shrimp are suitable ecotoxicological proxies for deep-sea hydrothermal vent shrimp the effects of waterborne Cu exposure (3 and 10 days at 0.4 and 4 µM concentrations) in Palaemon elegans, Palaemon serratus, and Palaemon varians were compared with Mirocaris fortunata. Accumulation of Cu and a set of biomarkers were analysed. Results show different responses among congeneric species indicating that it is not appropriate to use shallow-water shrimps as ecotoxicological proxies for deep-water shrimps. During the evolutionary history of these species they were likely subject to different chemical environments which may have induced different molecular/biochemical adaptations/tolerances. Results highlight the importance of analysing effects of deep-sea mining in situ and in local species to adequately assess ecotoxicological effects under natural environmental conditions.

In situ study of single-walled carbon nanotube growth in an environmental scanning electron microscope.

Monitoring individual single-walled carbon nanotubes (SWCNTs) during their growth is a highly sought-after goal in view of understanding the processes involved in the nucleation, elongation and termination which ultimately control the diameter and chiral selectivity. Here, we report on the first truly in situ observations of SWCNT growth in an environmental scanning electron microscope (ESEM). The CNT growth from lithographically patterned catalysts was investigated as a function of the catalyst type (Fe, Co or Ni), temperature, type of precursor (ethanol or acetylene), gas phase composition and pressure, and pretreatment conditions, and we report on the most appropriate conditions for SWCNT growth in ESEM conditions. We show that this approach allows the observation at the submicron scale of the different steps of the nanotube synthesis including the catalyst reduction, the growth and percolation of the nanotube network, and the deposition of individual nanotubes grown in the gas phase on the substrate. Despite these obvious advantages, we identified a few limitations which will need to be tackled for fully taking advantage of the approach, for instance for monitoring the growth of individual SWCNTs by ESEM, including the short lifetime of the catalyst nanoparticles, the preference for kite growth (by opposition to surface growth) and the influence of the electron beam on the nanotube growth.

Development of an ecotoxicological protocol for the deep-sea fauna using the hydrothermal vent shrimp Rimicaris exoculata.

In light of deep-sea mining industry development, particularly interested in massive-sulphide deposits enriched in metals with high commercial value, efforts are increasing to better understand potential environmental impacts to local fauna. The aim of this study was to assess the natural background levels of biomarkers in the hydrothermal vent shrimp Rimicaris exoculata and their responses to copper exposure at in situ pressure (30MPa) as well as the effects of depressurization and pressurization of the high-pressure aquarium IPOCAMP. R. exoculata were collected from the chimney walls of the hydrothermal vent site TAG (Mid Atlantic Ridge) at 3630m depth during the BICOSE cruise in 2014. Tissue metal accumulation was quantified in different tissues (gills, hepatopancreas and muscle) and a battery of biomarkers was measured: metal exposure (metallothioneins), oxidative stress (catalase, superoxide dismutase, glutathione-S-transferase and glutathione peroxidase) and oxidative damage (lipid peroxidation). Data show a higher concentration of Cu in the hepatopancreas and a slight increase in the gills after incubations (for both exposed groups). Significant induction of metallothioneins was observed in the gills of shrimps exposed to 4µM of Cu compared to the control group. Moreover, activities of enzymes were detected for the in situ group, showing a background protection against metal toxicity. Results suggest that the proposed method, including a physiologically critical step of pressurizing and depressurizing the test chamber to enable the seawater exchange during exposure to contaminants, is not affecting metal accumulation and biomarkers response and may prove a useful method to assess toxicity of contaminants in deep-sea species.

The potential for climate-driven bathymetric range shifts: sustained temperature and pressure exposures on a marine ectotherm, Palaemonetes varians.

Range shifts are of great importance as a response for species facing climate change. In the light of current ocean-surface warming, many studies have focused on the capacity of marine ectotherms to shift their ranges latitudinally. Bathymetric range shifts offer an important alternative, and may be the sole option for species already at high latitudes or those within enclosed seas; yet relevant data are scant. Hydrostatic pressure (HP) and temperature have wide ranging effects on physiology, importantly acting in synergy thermodynamically, and therefore represent key environmental constraints to bathymetric migration. We present data on transcriptional regulation in a shallow-water marine crustacean (Palaemonetes varians) at atmospheric and high HP following 168-h exposures at three temperatures across the organisms' thermal scope, to establish the potential physiological limit to bathymetric migration by neritic fauna. We observe changes in gene expression indicative of cellular macromolecular damage, disturbances in metabolic pathways and a lack of acclimation after prolonged exposure to high HP. Importantly, these effects are ameliorated (less deleterious) at higher temperatures, and exacerbated at lower temperatures. These data, alongside previously published behavioural and heat-shock analyses, have important implications for our understanding of the potential for climate-driven bathymetric range shifts.

Acute combined pressure and temperature exposures on a shallow-water crustacean: novel insights into the stress response and high pressure neurological syndrome.

Little is known about the ecological and physiological processes governing depth distribution limits in species. Temperature and hydrostatic pressure are considered to be two dominant factors. Research has shown that some marine ectotherms are shifting their bathymetric distributions in response to rapid anthropogenic ocean surface warming. Shallow-water species unable to undergo latitudinal range shifts may depend on bathymetric range shifts to seek refuge from warming surface waters. As a first step in constraining the molecular basis of pressure tolerance in shallow water crustaceans, we examined differential gene expression in response to acute pressure and temperature exposures in juveniles of the shallow-water shrimp Palaemonetes varians. Significant increases in the transcription of genes coding for an NMDA receptor-regulated protein, an ADP ribosylation factor, ß-actin, two heat shock protein 70 kDa isoforms (HSP70), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were found in response to elevated pressure. NMDA receptors have been implicated in pathways of excitotoxic damage to neurons and the onset of high pressure neurological syndrome (HPNS) in mammals. These data indicate that the sub-lethal effects of acute barotrauma are associated with transcriptional disturbances within the nervous tissue of crustaceans, and cellular macromolecular damage. Such transcriptional changes lead to the onset of symptoms similar to that described as HPNS in mammals, and may act as a limit to shallow water organisms' prolonged survival at depth.

Surface decoration of catanionic vesicles with superparamagnetic iron oxide nanoparticles: a model system for triggered release under moderate temperature conditions.

We report the design of new catanionic vesicles decorated with iron oxide nanoparticles, which could be used as a model system to illustrate controlled delivery of small solutes under mild hyperthermia. Efficient release of fluorescent dye rhodamine 6G was observed when samples were exposed to an oscillating magnetic field. Our system provides direct evidence for reversible permeability upon magnetic stimulation.

How to explain the difficulties in the coffinite synthesis from the study of uranothorite?

The preparation of Th(1-x)U(x)SiO(4) uranothorite solid solutions was successfully undertaken under hydrothermal conditions (T = 250 °C). From XRD and EDS characterization, the formation of a complete solid solution between x = 0 (thorite) and x = 0.8 was evidenced. Nevertheless, additional (Th,U)O(2) dioxide and amorphous silica were systematically observed for the highest uranium mole loadings. The influence of kinetics parameters was then studied to avoid the formation of such side products. The variation of the synthesis duration allowed us to point out the initial formation of oxide phases then their evolution to a silicate phase through a dissolution/precipitation process close to that already described as coffinitization. Also, the uranium mole loading initially considered was found to significantly influence the kinetics of reaction, as this latter strongly slows down for x > 0.3. Under these conditions, the difficulties frequently reported in the literature for the synthesis of pure USiO(4) coffinite were assigned to a kinetic hindering associated with the coffinitization reaction.

Stability and structural evolution of Ce(IV)(1-x)Ln(III)(x)O(2-x/2) solid solutions: a coupled µ-Raman/XRD approach.

Several CeO(2)-based mixed oxides with general composition Ce(1-x)Ln(x)O(2-x/2) (for 0 ≤ x ≤ 1 and Ln = La, Nd, Sm, Eu, Gd, Dy, Er, or Yb) were prepared using an initial oxalic precipitation leading to a homogeneous distribution of cations in the oxides. After characterization of the Ce/Nd oxalate precursors and then thermal conversion to oxides at T = 1000 °C, investigation of the crystalline structure of these oxides was carried out by XRD and µ-Raman spectroscopy. Typical fluorite Fm ̅3m structure was obtained for relatively low Ln(III) contents, while a cubic Ia ̅3̅ superstructure was evidenced above x ≈ 0.4. Moreover, since Nd(2)O(3) does not crystallize with the Ia ̅3̅-type structure, two-phase systems composed with additional hexagonal Nd(2)O(3) were obtained for x(Nd) ≥ 0.73 in the Ce(1-x)Nd(x)O(2-x/2) series. The effect of heat treatment temperature on these limits was explored through µ-Raman spectroscopy, which allowed determining the presence of small amounts of the different crystal structures observed. In addition, the variation of the Ce(1-x)Ln(x)O(2-x/2) unit cell parameter was found to follow a quadratic relation as a result of the combination between increasing cationic radius, modifications of cation coordination, and decreasing O-O repulsion caused by oxygen vacancies.

Effects of cold-exposure and subsequent recovery on cellular proliferation with influence of 20-hydroxyecdysone in a lepidopteran cell line (IAL-PID2).

In developing insects, the peak level of 20-hydroxyecdysone (20E) initiates a decrease in cyclin expression, which subsequently triggers an arrest of cellular proliferation and the start of differentiation, finally culminating in the moult. We investigated the impact of cold-exposure (4 degrees C) and recovery (26 degrees C) on the cell cycle activity of the Plodia interpunctella Lepidoptera cell line IAL-PID2 and on the expression of B-type cyclin (PcycB), ecdysone receptor (B1-isoform; PiEcR-B1), and Hsc70 (PiHsc70) mRNA. Cold-exposure significantly reduced expression of these mRNAs, while their levels increased to above control values during subsequent recovery at the normal growth temperature. When cold-exposed cells were returned to 26 degrees C, cell cycle activity restarted, but apoptosis was strongly increased. The presence of 20E appeared to increase this apoptotic phenomenon. This result is consistent with the described protective role of 20E against a variety of stressors and with the capacity of 20E to induce cell death in different situations. Here, we illustrate for the first time a connection between 20E treatment and Hsc70 expression during cold-exposure and subsequent recovery in insect cells. Combined with the 20E-induced apoptotic response, our results suggest that regulation of Hsc70 expression by 20E could act in synergy with the control of apoptotic cell death in order to optimize the survival of specific cell populations after a period of cold-exposure.

Sequence analysis of the myosin regulatory light chain gene of the vestimentiferan Riftia pachyptila.

We have isolated and characterized a cDNA (DNA complementary to RNA) clone (Rf69) from the vestimentiferan Riftia pachyptila. The cDNA insert consists of 1169 base pairs. The aminoacid sequence deduced from the longest reading frame is 193 residues in length, and clearly characterized it as a myosin regulatory light chain (RLC). The RLC primary structure is described in relation to its function in muscle contraction. The comparison with other RLCs suggested that Riftia myosin is probably regulated through its RLC either by phosphorylation like the vertebrate smooth muscle myosins, and/or by Ca2+-binding like the mollusk myosins. Riftia RLC possesses a N-terminal extension lacking in all other species besides the earthworm Lumbricus terrestris. Aminoacid sequence comparisons with a number of RLCs from vertebrates and invertebrates revealed a relatively high identity score (64%) between Riftia RLC and the homologous gene from Lumbricus. The relationships between the members of the myosin RLCs were examined by two phylogenetic methods, i.e. distance matrix and maximum parsimony. The resulting trees depict the grouping of the RLCs according to their role in myosin activity regulation. In all trees, Riftia RLC groups with RLCs that depend on Ca2+-binding for myosin activity regulation.

A novel chitin-binding protein from the vestimentiferan Riftia pachyptila interacts specifically with beta-chitin. Cloning, expression, and characterization.

A cDNA from Riftia pachyptila was cloned. It encodes a novel 21.3-kDa protein from the worm protective tube, named RCBP (for Riftia chitin-binding protein). On the basis of partial tube-peptide sequences previously obtained, experiments using reverse transcriptase-mediated polymerase chain reaction and rapid amplification of cDNA ends led to the complete cDNA sequence. Analysis of its deduced amino acid sequence shows the presence of two chitin-binding domains. These domains are closely related to type 2 chitin-binding domains that are restricted to the animal kingdom. We showed by affinity assay and immunogold labeling that RCBP is the first protein so far known that binds specifically beta-chitin and that is unable to bind the most common alpha-form found in chitin secreting animals. The RCBP mRNA was found to be present in specific epidermal cells from the worm body wall, but never in the chitin-secreting gland cells. This unexpected result clearly indicates that this tube protein is synthesized in specialized areas of the outer epithelium and that at least two different tissues are involved in this exoskeleton synthesis.