Sex-specific seasonal variations in the fatty acid and carotenoid composition of sea cucumber gonads and implications for aquaculture.

Fatty acids and carotenoids are known to have roles in embryonic and larval development of sea cucumbers, but their changes in gonads during gametogenesis have not yet been studied. To improve our knowledge of the reproductive cycle of sea cucumbers in an aquaculture perspective, we collected 6-11 individuals of the species Holothuria (Panningothuria) forskali Delle Chiaje, 1823 approximately every 2 months from December 2019 to July 2021 east of the Glenan Islands (Brittany - France; 47.710°N, 3.948°W) at a depth of 8-12 m. Our results show that soon after spawning, sea cucumbers take advantage of an increased food availability in spring to rapidly and opportunistically accumulate nutrients in the form of lipids in their gonads (from May to July) and then slowly elongate, desaturate and probably rearrange fatty acids within lipid classes for the next reproductive season according to the specific requirements of both sexes. In contrast, acquisition of carotenoids occurs synchronously with gonads filling and/or through the reabsorption of spent tubules (T5), thus revealing little seasonal variations at the scale of the entire gonad in terms of relative abundance in both sexes. All results suggest that gonads are fully replenished with nutrients by October and that broodstock for induced reproduction could be captured at this moment and kept until the production of larvae is required. Maintaining broodstock for consecutive years would probably be a higher level challenge as the dynamics of tubule recruitment are not fully understood and seem to last for several years. Supplementary Information: The online version contains supplementary material available at 10.1007/s00227-023-04198-0.

Responses of early life stages of European abalone (Haliotis tuberculata) to ocean acidification after parental conditioning: Insights from a transgenerational experiment.

CO2 absorption is leading to ocean acidification (OA), which is a matter of major concern for marine calcifying species. This study investigated the effects of simulated OA on the reproduction of European abalone Haliotis tuberculata and the survival of its offspring. Four-year-old abalone were exposed during reproductive season to two relevant OA scenarios, ambient pH (8.0) and low pH (7.7). After five months of exposure, abalone were induced to spawn. The gametes, larvae and juveniles were then exposed for five months to the same pH conditions as their parents. Several biological parameters involved in adult reproduction as well as in larval, post-larval and juvenile fitness were measured. No effects on gametes, fertilisation or larval oxidative stress response were detected. However, developmental abnormalities and significant decreases in shell length and calcification were observed at veliger stages. The expression profile of a GABA A receptor-like gene appeared to be regulated by pH, depending on larval stage. Larval and post-larval survival was not affected by low pH. However, a lower survival and a reduction of growth were recorded in juveniles at pH 7.7. Our results confirm that OA negatively impacts larval and juvenile fitness and suggest the absence of carry-over effects on abalone offspring. This may compromise the survival of abalone populations in the near future.

New insight on spatial localization and microstructures of calcite-aragonite interfaces in adult shell of Haliotis tuberculata: Investigations of wild and farmed abalones by FTIR and Raman mapping.

In the present study, we investigated the shell microstructures of the gastropod European abalone Haliotis tuberculata in order to clarify the complex spatial distribution of the different mineral phases. Our studies were carried out with a standardized methodology on thirty adult European abalone H. tuberculata (5-6 cm long) composed of 15 wild individuals and 15 individuals taken from the France Haliotis hatchery. The macroscopic (binocular) and microscopic observations coupled with Fourier Transform Infrared Spectroscopy (FTIR) and Raman vibrational analysis allowed to unambiguously detect, identify and localize calcite and aragonite. For the first time it has been shown that calcite is present in 100% of farmed and wild adult shell. The microstructural details of the calcite-aragonite interfaces were revealed by using both confocal micro-Raman mapping and Scanning Electron Microscopy (SEM) observations. Calcite zones are systematically found in the spherulitic layer without direct contact with the nacreous layer. The calcite area - nacreous layer interface is made of a thin spherulitic layer with variable thickness from a few micrometers to several millimeters. In order to contribute to a better understanding of the biomineralization process, a model explaining the hierarchical arrangement of the different phases of calcium carbonate is presented and discussed. Finally, it has been shown that these calcitic zones can be connected to each other within the shells and that their spatial distributions correspond to streaks perpendicular to the direction of length growth.

Combined effects of ocean warming and acidification on the larval stages of the European abalone Haliotis tuberculata.

This study examined the physiological responses of the larval stages of Haliotis tuberculata, an economically important abalone, to combined temperature (17 °C and 19 °C) and pH (ambient pH and -0.3 units, i.e., +200% increase in seawater acidity) in a full factorial experiment. Tissue organogenesis, shell formation, and shell length significantly declined due to low pH. High temperature significantly increased the proportion of fully shelled larvae at 24 h post-fertilization (hpf), but increased the proportion of unshelled larvae at 72 hpf. Percentage of swimming larvae at 24 hpf, 72 hpf and 96 hpf significantly declined due to high temperature, but not because of low pH. Larval settlement increased under high temperature, but was not affected by low pH. Despite the fact that no interaction between temperature and pH was observed, the results provide additional evidence on the sensitivity of abalone larvae to both low pH and high temperature. This may have negative consequences for the persistence of abalone populations in natural and aquaculture environments in the near future.

Acid-base balance in the hæmolymph of European abalone (Haliotis tuberculata) exposed to CO2-induced ocean acidification.

Ocean acidification (OA) and the associated changes in seawater carbonate chemistry pose a threat to calcifying organisms. This is particularly serious for shelled molluscs, in which shell growth and microstructure has been shown to be highly sensitive to OA. To improve our understanding of the responses of abalone to OA, this study investigated the effects of CO2-induced ocean acidification on extra-cellular acid-base parameters in the European abalone Haliotis tuberculata. Three-year-old adult abalone were exposed for 15 days to three different pH levels (7.9, 7.7, 7.4) representing current and predicted near-future conditions. Hæmolymph pH and total alkalinity were measured at different time points during exposure and used to calculate the carbonate parameters of the extracellular fluid. Total protein content was also measured to determine whether seawater acidification influences the composition and buffer capacity of hæmolymph. Extracellular pH was maintained at seawater pH 7.7 indicating that abalones are able to buffer moderate acidification (-0.2 pH units). This was not due to an accumulation of HCO3- ions but rather to a high hæmolymph protein concentration. By contrast, hæmolymph pH was significantly decreased after 5 days of exposure to pH 7.4, indicating that abalone do not compensate for higher decreases in seawater pH. Total alkalinity and dissolved inorganic carbon were also significantly decreased after 15 days of low pH exposure. It is concluded that changes in the acid-base balance of the hæmolymph might be involved in deleterious effects recorded in adult H. tuberculata facing severe OA stress. This would impact both the ecology and aquaculture of this commercially important species.

Comparative ultrastructure and carbohydrate composition of gastroliths from astacidae, cambaridae and parastacidae freshwater crayfish (crustacea, decapoda).

Crustaceans have to cyclically replace their rigid exoskeleton in order to grow. Most of them harden this skeleton by a calcification process. Some decapods (land crabs, lobsters and crayfish) elaborate calcium storage structures as a reservoir of calcium ions in their stomach wall, as so-called gastroliths. For a better understanding of the cyclic elaboration of these calcium deposits, we studied the ultrastructure of gastroliths from freshwater crayfish by using a combination of microscopic and physical techniques. Because sugars are also molecules putatively involved in the elaboration process of these biomineralizations, we also determined their carbohydrate composition. This study was performed in a comparative perspective on crayfish species belonging to the infra-order Astacidea (Decapoda, Malacostraca): three species from the Astacoidea superfamily and one species from the Parastacoidea superfamily. We observed that all the gastroliths exhibit a similar dense network of protein-chitin fibers, from macro- to nanoscale, within which calcium is precipitated as amorphous calcium carbonate. Nevertheless, they are not very similar at the molecular level, notably as regards their carbohydrate composition. Besides glucosamine, the basic carbohydrate component of chitin, we evidenced the presence of other sugars, some of which are species-specific like rhamnose and galacturonic acid whereas xylose and mannose could be linked to proteoglycan components.

Detailed FT near-infrared study of the behaviour of water and hydroxyl in sedimentary length-fast chalcedony, SiO2, upon heat treatment.

Chalcedony is a porous spatial arrangement of hydroxylated nanometre sized α-quartz (SiO(2)) crystallites. Due to micro-structural transformations upon heat treatment, the optical and mechanical properties of the rock are modified. We investigated these transformations in sedimentary length-fast chalcedony through Fourier Transform near- and mid-infrared spectroscopy using direct transmission and the reflectivity. Chemical adsorption potential and absorption of H(2)O by pores was studied after heat treatment. We found that water held in open porosity is reduced upon heat treatment to temperatures above 150°C. Silanole is noticeably lost from 250 to 300°C upwards and new bridging Si-O-Si further reduces the surface of open pores, creating a less porous material. Molecular water, resulting from the reaction Si-OH HO-Si→Si-O-Si+H(2)O creates new isolated pores within the material. At temperatures above 500°C, the samples start internal fracturing, permitting water held in isolated pores to be evacuated. These results shed light on thermal transformations in chalcedony and allow for a better understanding of mechanical transformations after heat treatment.

Ultrastructure, chemistry and mineralogy of the growing shell of the European abalone Haliotis tuberculata.

An integrated study of shell formation was initiated covering the entire life cycle of the marine gastropod Haliotis tuberculata. Shell microstructure, chemistry and mineralogy were investigated by polarized microscopy, scanning electron microscopy (SEM), energy dispersive X-ray spectrometry (EDX) and infra-red (IR) spectroscopy. SEM images of trochophore and veliger larvae showed the different stages of shell growth from the initial shell field to the late calcified protoconch. Cross-sections revealed the microstructural arrangement of biominerals, showing the progressive mineralization of the organic protoconch prior to metamorphosis. To gain more information on mineralogical composition, EDX analyses and IR spectroscopy were performed along the development stages. The results demonstrated that early protoconch was mostly composed of amorphous calcium carbonate, while veliger stages showed a gradually crystallization under the form of aragonite. Post-metamorphic shell contained two distinct parts, the original protoconch supporting the new juvenile shell characterized by a marked sculptural pattern. The shells from post-larval and juvenile abalones were essentially made of aragonite.