Renata Boucher-Rodoni

Bioaccumulation of 12 Trace Elements in the Tissues of the Nautilus Nautilus macromphalus from New Caledonia

Soils in New Caledonia are particularly rich in metals among which Fe and Ni are intensively exploited. Due to important natural erosion in tropical latitudes and to mining activities, coastal waters are enriched in Co, Cr, Fe and Ni. In deeper waters lives a cephalopod species which is considered as a living fossil, the nautilus Nautilus macromphalus. In this study, 12 trace elements were analysed in the tissues of 4 nautilus specimens. Results showed high metal concentrations compared to data available for cephalopods from temperate waters. These concentrations were often in the same order as those encountered in bivalves or gastropods from contaminated areas. Relatively high concentrations of Ni and Cr in the haemolymph strongly suggest a high exposure of N. macromphalus to these metals. Among the tissues, the digestive gland has the highest concentrations of Cd, Co, Fe, V and Zn while for Ag, Al, As, Cr and Ni, renal and pericardial appendages exhibited the highest values. Despite this, the digestive gland contained the largest quantities of all metals with the exception of As and Mn which were mainly found in the body muscular remains. These results highlighted the major role of digestive gland and excreting organs in the metabolism of metals in these cephalopod species.

Comparative bioaccumulation of trace elements between Nautilus pompilius and Nautilus macromphalus (Cephalopoda: Nautiloidea) from Vanuatu and New Caledonia

The concentrations of 16 trace elements were investigated and compared for the first time in the digestive and excreting tissues of two Nautilus species (Cephalopoda: Nautiloidea) from two geologically contrasted areas: (1) N. macromphalus from New Caledonia, a region characterized by its richness in nickel ores and its lack of tectonic activities and (2) N. pompilius from the Vanuatu archipelago showing high volcanic and tectonic activities. In both Nautilus species, results clearly highlighted that the digestive gland played a key role in the bioaccumulation and storage of Ag, Cd, Ce, Co, Cu, Fe, La, Nd, V, and Zn whereas As, Cr, Mn, Ni, Pb, and Se were accumulated in a greater extent in the excreting tissues (i.e. pericardial and renal appendages). Despite contrasting environments, no significant difference (p<0.05) was found between the two Nautilus species in the concentrations of most of the essential and non-essential elements, including Ni and associated metals in Ni ores (i.e. Co and Mn). As nautilus lives on the outer shelf of barrier reefs, these results strongly support the hypothesis that the New Caledonian lagoon traps the major amount of the trace elements derived from natural erosion and the intense mining activities conducted on land. In contrast, the concentrations of the rare earth elements (Ce, La, and Nd) were significantly higher in N. pompilius than in N. macromphalus, probably as a result of the local enrichment of Vanuatu waters by specific environmental processes, such as volcanism or upwelling.

Respiration and nitrogen excretion by the squid Loligo forbesi

Respiration and nitrogen excretion rates of mature adult Loligo forbesi were investigated at the Roscoff Laboratory (North Brittany, France) during individual short-term incubation experiments in January 1986. The squids were in post-digestive condition and not actively swimming. Both oxygen uptake and nitrogen excretion are continuous processes. The metabolic rates of this active nektonic species (145 ml kg-1 h-1 oxygen uptake, 18.56 μg g-1 h-1 ammonia excretion) are distinctly higher than those of benthic cephalopods. Proteins constitute the main metabolic substrate for energetic needs. Besides ammonia, urea is also continuously released, in amounts ranging from 5 to 16% of ammonia-excretion values.

Enigmatic dual symbiosis in the excretory organ of Nautilus macromphalus (Cephalopoda: Nautiloidea)

Symbiosis is an important driving force in metazoan evolution and the study of ancient lineages can provide an insight into the influence of symbiotic associations on morphological and physiological adaptations. In the ‘living fossil’ Nautilus, bacterial associations are found in the highly specialized pericardial appendage. This organ is responsible for most of the excretory processes (ultrafiltration, reabsorption and secretion) and secretes an acidic ammonia-rich excretory fluid. In this study, we show that Nautilus macromphalus pericardial appendages harbour a high density of a β-proteobacterium and a coccoid spirochaete using transmission electron microscopy, comparative 16S rRNA sequence analysis and fluorescence in situ hybridization (FISH). These two bacterial phylotypes are phylogenetically distant from any known bacteria, with ammonia-oxidizing bacteria as the closest relatives of the β-proteobacterium (above or equal to 87.5% sequence similarity) and marine Spirochaeta species as the closest relatives of the spirochaete (above or equal to 89.8% sequence similarity), and appear to be specific to Nautilus. FISH analyses showed that the symbionts occur in the baso-medial region of the pericardial villi where ultrafiltration and reabsorption processes take place, suggesting a symbiotic contribution to the excretory metabolism.

In situ study of the effect of oyster biomass on benthic metabolic exchange rates

Oxygen consumption and dissolved nitrogen fluxes at the water-sediment interface of an oyster-bed were measured in situ using transparent enclosures inserted on undisturbed sediment. Experiments were performed in summer, under dark and light conditions, with various densities of the oyster Crassostrea gigas (0–150 animals m − 2). The influence of oyster biomass on oxygen and ammonia exchange rates was similar in both lighting conditions. Oxygen consumption increased with increasing biomass, though not at the level of prediction for the highest biomasses, suggesting a depressed respiration rate. Ammonia release never matched the rates predicted by adding sediment efflux to oyster excretion, when biomass exceeded 100 g DW m − 2. The coupling between oxygen consumption and ammonia release (O : N ratio) was thus influenced by oyster biomass. Stabilization of nitrogen release was related to enhanced nitrification in the presence of oyster and/or sediment uptake of ammonia against the molecular gradient. Urea release was erratic but appeared uninfluenced by oyster biomass. Fluxes of oxygen and of each nitrogen compound displayed thus a specific response to modifications of the oyster biomass. Both the organisms and the sediment are involved in regulation processes of metabolic exchange rates at the water-sediment interface.

Respiratory quotient and calcification of Nautilus macromphalus (Cephalopoda: Nautiloidea)

Respiration and calcification were investigated in the ectocochleate cephalopod Nautilus macromphalus Sowerby. Specimens were collected off New Caledonia, in October 1991, and kept at the Nouméa Aquarium until December 1991. The respiratory quotient and calcification rate of 5 individuals were measured during 14 short term incubations (63 to 363 min). Oxygen uptake was recorded with a polarographic oxygen sensor. CO2 flux and calcification were calculated from changes in pH and alkalinity (alkalinity-anomaly technique). Several methods were used to compute the respiratory quotient (RQ); a functional regression indicated an RQ of 0.74. CaCO3 exchange rates were linearly related to respiratory quotient, calcification occurring in individuals with a low RQ. CaCO3 uptake from the surrounding water was noncontinuous. From the highest CaCO3 uptake, maximum growth rate was estimated as 7.1 mg shell wt h- (=61 g yr-1).

Fluctuations des nutriments au cours de la marée sur les parcs Ostreicoles de la rivière Penze (Nord-Finistère)

The importance of short-term fluctuations in nutrients and primary amines over oyster beds during spring and neap tidal cycles was analysed. The range of the variations recorded over 24 h is of the same order of magnitude as that observed over an annual cycle.Multivariate analysis, taking into account all available parameters concerning the tidal cycles, showed a short-term evolution of primary amines and ammonia nitrogen, independent from that of other parameters. The latter fluctuate in synchronisation with the down-and-up-stream flow of the river Penzé. The mean ammonia Nitrogen concentration is higher during spring tide than during neap tide, and ammonia is more concentrated at the bottom than in the surface water. This higher concentration of ammonia in bottom water is related neither to current speed, nor to interstitial water dilution by sediment resuspension. Nutrient exchanges between the sediment and the water column did not seem to be an important source of ammonia, while ammonia excretion by oysters might be the main source of this nutrient at the bottom level, particularly during spring tide.

Identification of a Vibrio strain producing antimicrobial agents in the excretory organs of Nautilus pompilius (Cephalopoda: Nautiloidea)

The aim of the present study was to identify and characterize bacteria producing antimicrobial compounds in the excretory organs of Nautilus pompilius. Culture-dependent and culture-independent complementary approaches were used for bacterial identification such as: culture on selective media, Gram staining, CARD-FISH, direct DNA extraction from host tissue, PCR amplification and sequencing of the bacterial 16S rRNA gene. Results show presence of three bacterial groups: γ-Proteobacteria with three clusters (Pseudomonadales, Vibrionales, Alteromonadales), β-Proteobacteria and spirochetes. In order to screen for active strains, antimicrobial activity was tested by diffusion agar assay against Micrococcus luteus, Escherichia coli, Vibrio harveyi, and Candida albicans. Nautilus isolates showed antimicrobial activities against both Gram-positive and Gram-negative reference strains. Most of the active strains were phylogenetically related to environmental Vibrionaceae. These strains were always abundant in N. pompilius PA but were absent from Nautilus macromphalus from other geographical areas. Therefore, we suggest that antimicrobial active Vibrionaceae infect N. pompilius by environmental transmission.

Microeledone mangoldi n. gen. and n. sp., a deep-water pygmy octopus from the Norfolk Ridge, New Caledonia (Cephalopoda : Octopodidae)

Microeledone mangoldi n. gen. and n. sp. is described from a male specimen collected from approximately one kilometre deep on the Norfolk Ridge south of New Caledonia in the south-west Pacific Ocean. This tiny octopus is characterised by a single row of suckers that are functional to the tips of the arms, the absence of an ink sac, the presence of a pair of chitinous stylets, a UU-shaped funnel organ, a copulatory organ with distinct ligula and calamus, and a distinctive radula. The radula has seven teeth per row. The unique central (rachidian) tooth has a long, sharp mesocone with a curved and grooved tip. The remaining rows of teeth are also unique, being flattened and plate-like. Marginal plates are absent. The body is smooth, without papillae, cartilaginous tubercles, or a lateral ridge. Functional chromatophores are absent. This octopus appears related to several other genera of deep-sea octopods with a single row of suckers, namely Thaumeledone Robson, 1930, Bentheledone Robson, 1932 and Graneledone Joubin, 1918, to which the new genus is compared. Definitions of the genus Thaumeledone are reviewed.

Occurrence of a specific dual symbiosis in the excretory organ of geographically distant Nautiloids populations

Nautilus is one of the most intriguing of all sea creatures, sharing morphological similarities with the extinct forms of coiled cephalopods that evolved since the Cambrian (542-488 mya). Further, bacterial symbioses found in their excretory organ are of particular interest as they provide a great opportunity to investigate the influence of host-microbe interactions upon the origin and evolution of an innovative nitrogen excretory system. To establish the potential of Nautilus excretory organ as a new symbiotic system, it is, however, necessary to assess the specificity of this symbiosis and whether it is consistent within the different species of present-day Nautiloids. By addressing the phylogeny and distribution of bacterial symbionts in three Nautilus populations separated by more than 6000 km (N. pompilius from Philippines and Vanuatu, and N. macromphalus from New Caledonia), this study confirms the specificity of this dual symbiosis involving the presence of betaproteobacteria and spirochaete symbionts on a very wide geographical area. Overall, this work sheds further light on Nautiloids excretory organ as an innovative system of interaction between bacteria and cephalopods.