The dark side of soft tissues: Unexpected inorganic carbonate in the invasive slipper limpet Crepidula fornicata and its implications for stable isotope interpretations.

RATIONALE: Stable isotopic analysis is extensively used in trophic ecology. Inorganic carbonates, usually originating from shell fragments, are routinely removed from samples using an acid treatment because they affect δ13 C values. However, acid treatment can also change δ15 N values. For some taxa such as molluscs, the general assumption is that acid pre-treatment is not necessary since their shell is easily dissected from soft tissues and represents the only source of inorganic carbonates. However, other sources of non-dietary carbon (e.g., intracellular inorganic carbon) enriched in13 C thus get overlooked. METHODS: Soft tissues (foot) of the invasive gastropod Crepidula fornicata of different size classes were analysed for their δ13 C and δ15 N values with and without acid pre-treatment using isotope ratio mass spectrometry. In toto microscopic investigations coupled with acid treatment, scanning electron microscopy and energy-dispersive spectroscopy were used to highlight the presence of inorganic carbonate. A correction model was derived and applied to existing stable isotope data for C. fornicata. We used both seasonal variations in δ13 C signatures and mixing model outputs to assess the error in δ13 C values. RESULTS: Acid pre-treatment had a significant effect on the stable isotope compositions of C. fornicata foot tissue, especially on δ13 C values: isotopic differences increased with size, up to 3‰ for large females. No effect was detected for small (below ~20 mm) and motile males. In toto microscopic analysis revealed the presence of small spherules of inorganic carbonate, hence explaining the differences in δ13 C values. Mixing model outputs and seasonal variation of δ13 C values showed that untreated samples can lead to large misinterpretations about diet proportions and degree of trophic niche overlap, respectively. CONCLUSIONS: Spherules of inorganic carbonate in C. fornicata soft tissues are likely to be linked with the motility of this species and their mucus production. We recommend assessing the presence of inorganic carbonate in soft tissue of sessile gastropods.

Multiple beneficial lipids including lecithin detected in the edible invasive mollusk Crepidula fornicata from the French Northeastern Atlantic coast.

The invasive mollusk Crepidula fornicata, occurring in large amounts in bays along the French Northeastern Atlantic coasts, may have huge environmental effects in highly productive ecosystems where shellfish are exploited. The present study aims at determining the potential economic value of this marine species in terms of exploitable substances with high added value. Lipid content and phospholipid (PL) composition of this mollusk collected on the Bourgneuf Bay were studied through four seasons. Winter specimens contained the highest lipid levels (5.3% dry weight), including 69% of PLs. Phosphatidylcholine (PC) was the major PL class all year, accounting for 63.9% to 88.9% of total PLs. Consequently, the winter specimens were then investigated for PL fatty acids (FAs), and free sterols. Dimethylacetals (DMAs) were present (10.7% of PL FA + DMA mixture) revealing the occurrence of plasmalogens. More than forty FAs were identified, including 20:5n-3 (9.4%) and 22:6n-3 (7.3%) acids. Fourteen free sterols were present, including cholesterol at 31.3% of the sterol mixture and about 40% of phytosterols. These data on lipids of C. fornicata demonstrate their positive attributes for human nutrition and health. The PL mixture, rich in PC and polyunsaturated FAs, offers an interesting alternative source of high value-added marine lecithin.

Extensive spatial impacts of oyster reefs on an intertidal mudflat community via predator facilitation.

Habitat engineers make strong and far-reaching imprints on ecosystem processes. In intertidal mudflats, the dominant primary producer, microphytobenthos (MPB), often forms high biomass patches around oyster reefs. We evaluate multiple hypotheses linking MPB with oyster reefs, including oyster biodeposition, meiofaunal grazing, and abiotic factors, aiming to help predict effects of reef removal or proliferation. We quantify spatial patterns of an Atlantic mudflat community and its environment around two large Crassostrea reefs before experimentally sacrificing one reef via burning. MPB biomass was enriched surrounding living oyster reefs although infaunal biomass and individual sizes were low. Structural equation modelling best supported the hypothesis that crab predation intensity, which decayed with distance from the reefs, locally freed MPB from grazing. Our results suggest that Crassostrea reef expansion may enrich local MPB patches and redirect trophic energy flows away from mudflat infauna, with potential implications for the sustainability of local fisheries and bird conservation.

Realistic environmental exposure to microplastics does not induce biological effects in the Pacific oyster Crassostrea gigas.

The aim of the present study was to evaluate the presence and potential toxic effects of plastic fragments (<400 µm) of polyethylene and polypropylene on the Pacific oyster Crassostrea gigas. Oysters were exposed to environmentally relevant concentrations (0, 0.008, 10, 100 µg of particles/L) during 10 days, followed by a depuration period of 10 days in clean seawater. Effects of microplastics were evaluated on the clearance rate of organisms, tissue alteration, antioxidant defense, immune alteration and DNA damage. Detection and quantification of microplastics in oyster's tissues (digestive gland, gills and other tissues) and biodeposits using infrared microscopy were also conducted. Microplastics were detected in oyster's biodeposits following exposure to all tested concentrations: 0.003, 0.006 and 0.05 particles/mg of biodeposits in oysters exposed to 0.008, 10 and 100 µg of particles/L, respectively. No significant modulation of biological markers was measured in organisms exposed to microplastics in environmentally relevant conditions.

Biofilm grazing in a higher vertebrate: the western sandpiper, Calidris mauri.

We show that a higher vertebrate can graze surficial intertidal biofilm, previously only considered a food source for rasping invertebrates and a few specialized fish. Using evidence from video recordings, stomach contents, and stable isotopes, we describe for the first time the grazing behavior of Western Sandpipers (Calidris mauri) and estimate that biofilm accounts for 45-59% of their total diet or 50% of their daily energy budget. Our finding of shorebirds as herbivores extends the trophic range of shorebirds to primary consumers and potential competitors with grazing invertebrates. Also, given individual grazing rates estimated at seven times body mass per day and flock sizes into the tens of thousands, biofilm-feeding shorebirds could have major impacts on sediment dynamics. We stress the importance of the physical and biological processes maintaining biofilm to shorebird and intertidal conservation.