Shell disease: abnormal conchiolin deposit in the abalone Haliotis tuberculata

International audienceShell disease in the abalone Haliotis tuberculata L. is characterized by a conchiolin deposit on the inner surface of the shell. The gross clinical signs appear similar to the Brown Ring Disease (BRD) of clams. BRD has been extensively described in clams and is known to be responsible for severe mortalities and the collapse of the clam aquaculture industry in western France. In the clam, it was found to be caused by the infection of the mantle by Vibrio tapetis. Brown protein deposits have been observed in various abalone species around the world; some of these have been associated with a fungal infection in New Zealand, but the ones described here are similar to bacterial infections observed in clams. Larger animals appeared to be more affected by the disease, and a positive correlation of the number of successive infections found in the shells with the level of infestation of the shell by borers suggests that boring polychaetes and sponges may be vectors of the disease, or that the parasite infestation may increase the susceptibility of the animal to this infection. There is no evidence, however, that this infection causes mortality in abalone

Growth of the European abalone (Haliotis tuberculata L.) in situ: Seasonality and ageing using stable oxygen isotopes

International audienceThe ormer, Haliotis tuberculata is the only European abalone species commercially exploited. The determination of growth and age in the wild is an important tool for fisheries and aquaculture management. However, the ageing technique used in the past in the field is unreliable. The stable oxygen isotope composition (18O/16O) of the shell depends on the temperature and oxygen isotope composition of the ambient sea water. The stable oxygen isotope technique, developed to study paleoclimatological changes in shellfish, was applied to three H. tuberculata specimens collected in north-west Brittany. For the specimens collected, the oxygen isotope ratios of the shell reflected the seasonal cycle in the temperature. From winter-to-winter cycles, estimates of the age and the annual growth increment, ranging from 13 to 55 mm per year were obtained. This study shows that stable oxygen isotopes can be a reliable tool for ageing and growth studies of this abalone species in the wild, and for validating other estimates

The impacts of handling and air exposure on immune parameters, gene expression, and susceptibility to vibriosis of European abalone Haliotis tuberculata.

International audienceWild or farmed abalone are regularly exposed to stressors, such as air exposure and handling. Immune and transcriptional responses as well as susceptibility to vibriosis of sexually mature or immature European abalone acclimated at 16 or 19 °C were determined following handling or air exposure. Hemocyte density and H2O2 production increased while hemocyte viability and phagocytic index decreased following handling. Air exposure induces a decrease of hemocyte density and phagocytic index. Measurement of the expression of genes implicated in general metabolic, immunological and stress responses in gills, foot-muscle and hemocytes by real time q-PCR suggested that both stressors lead to a metabolic rate depression, characterized by a general inhibition of transcription. Finally, following handling a Vibrio harveyi challenge enhances almost 100% mortality of sexually immature animals at 19 °C while it has been previously demonstrated that only mature are susceptible to vibriosis

Summer immune depression associated with increased susceptibility of the European abalone, Haliotis tuberculata to Vibrio harveyi infection

International audienceHaliotis tuberculata mortality outbreaks have occurred in France since 1998 and were attributed to a pathogenic Vibrio harveyi. These mortalities were recorded in September, a month with abalone reproduction and characterised by high seawater temperatures. The importance of gonadal maturation and temperature increase on abalone immunity and susceptibility to V. harveyi infection needed to be clarified. Therefore, an immune survey analyzing a large panel of parameters was performed from June to September 2007 on abalone from the Bay of Brest. The data obtained were put in relation with abalone reproductive status and its susceptibility to V. harveyi. Most parameters showed clear patterns from early to late summer and during gametogenesis, phagocytosis and phenoloxidase activity were reduced, whereas basal reactive oxygen species production and agglutination titres were significantly increased. Total haemocyte counts went up after the partial spawning event at the end of June, and cell complexity diminished. Using a Principal Component Analysis, the "haemolymph profile" was shown to decrease in parallel with spawning and gonadal maturation processes, and reached a minimum just after total spawning. A significant correlation between this "haemolymph profile" and disease susceptibility allowed us to establish for the first time in abalone, a clear concordance between maturation and spawning processes, immune status and abalone susceptibility to V. harveyi

Construction of a stable GFP-tagged Vibrio harveyi strain for bacterial dynamics analysis of abalone infection

International audienceVibrio harveyi is a bacterial marine pathogen that can cause fatal disease in a large range of vertebrates and invertebrates, including the commercially important marine gastropod, Haliotis tuberculata. Since 1997, strains of this bacterium have regularly been causing high mortalities in farmed and wild abalone populations. The way in which the pathogen enters into abalone and the disease transmission mechanisms are thus far unknown. Therefore, a pathogenic strain, ORM4, was green fluorescent protein-tagged and validated both for its growth characteristics and for its virulence as a genuine model for abalone disease. The strain allows V. harveyi quantification by flow cytometry in seawater and in abalone haemolymph as well as the in situ detection of the parasite inside abalone tissues

Domestication modifies behaviour of first generation of domesticated abalone

The domestication of Haliotis tuberculata began recently. During this domestication process, abalone may acquire behavioural and physiological traits to become more adapted to their captive environment. These modifications could be the result of intentional selection on production traits, or of unintentional selection due to specific conditions experienced in the farm environment. In order to study this process, progenies of 3 different broodstock origins were studied. Wild abalone, farmed abalone selected for faster growth at 5% selection pressure and randomly selected farmed abalone were used as broodstock. Farmed abalone were the third generation of abalone resulting from systematic mating between wild and farmed broodstock (either males or females were wild broodstock), without intentional selection. Spawning was induced at 4 different periods, with different broodstock individuals used in the 3 treatments each time. After a 16 month rearing in individual tanks for each spawning (n= 12 tanks in total), offspring from the 3 progenies were individually tagged and placed together in sea‐cages at a density of 75 abalone per cage. Three replicates were used for each spawning period. At the age of 3 years in June 2017, mortality, growth, gonadic development, immune status (phagocytosis efficiency and total haemocyte count) as well as abalone behaviour in different situations (circadian, righting, predator and hiding behaviour) were studied. No differences were observed in term of survival, growth and physiological traits between the 3 progenies. Behavioural and dissection measures are still being analysed. These results suggest that the first stages of selection of H. tuberculata did not induce a significant modification of growth and physiology. Further behavioural analysis will be conducted to confirm this lack of effect

Stress response of farmed European abalone reveals rapid domestication process in absence of intentional selection

WOS:000413795800002International audienceFarming, and thus the domestication of Haliotis tuberculata, began recently. We compared the responses of unselected farmed and wild abalone to stressors that occur on farms. The aim was to determine if the farm environment had induced differences in the behavioural or physiological performances of the abalone. Thirty hatchery-born 3.5 year-old abalone and thirty wild ones were reared under standard farm conditions for 6 months and characterised for 19 traits related to growth, survival, behaviour and immunology. Behavioural and immunological responses to stressors differed between the two stocks. Farmed abalone retracted and swivelled less in reaction to a finger contact. Phagocytosis efficiency was reduced by shaking in abalone from both origins, but the farmed stock returned to the basal level after the recovery week, while wild abalone did not, and a rise of total haemocyte count after shaking and its return to a basal level after one week was only observed for the farmed stock. This indicates that both behaviour and immune responses following a stress have been affected by the farming practices. This suggests that a domestication process has already been initiated in the farmed stock. Our results may also be important for the success of any population enhancement based on hatchery-produced abalone as they raise the question of the capacity of abalone with a fanned origin to be adapted to the wild environment

Biomineralization markers during early shell formation in the European abalone Haliotis tuberculata

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Characterization of abalone Haliotis tuberculata-Vibrio harveyi interactions in gill primary cultures

The decline of European abalone Haliotis tuberculata populations has been associated with various pathogens including bacteria of the genus Vibrio. Following the summer mortality outbreaks reported in France between 1998 and 2000, Vibrio harveyi strains were isolated from moribund abalones, allowing in vivo and in vitro studies on the interactions between abalone H. tuberculata and V. harveyi. This work reports the development of primary cell cultures from abalone gill tissue, a target tissue for bacterial colonisation, and their use for in vitro study of host cell—V. harveyi interactions. Gill cells originated from four-day-old explant primary cultures were successfully sub-cultured in multi-well plates and maintained in vitro for up to 24 days. Cytological parameters, cell morphology and viability were monitored over time using flow cytometry analysis and semi-quantitative assay (XTT). Then, gill cell cultures were used to investigate in vitro the interactions with V. harveyi. The effects of two bacterial strains were evaluated on gill cells: a pathogenic bacterial strain ORM4 which is responsible for abalone mortalities and LMG7890 which is a nonpathogenic strain. Cellular responses of gill cells exposed to increasing concentrations of bacteria were evaluated by measuring mitochondrial activity (XTT assay) and phenoloxidase activity, an enzyme which is strongly involved in immune response. The ability of gill cells to phagocyte GFP-tagged V. harveyi was evaluated by flow cytometry and gill cells-V. harveyi interactions were characterized using fluorescence microscopy and transmission electron microscopy During phagocytosis process we evidenced that V. harveyi bacteria induced significant changes in gill cells metabolism and immune response. Together, the results showed that primary cell cultures from abalone gills are suitable for in vitro study of host-pathogen interactions, providing complementary assays to in vivo experiments

Relationships between growth, survival, physiology and behaviour — A multi-criteria approach to Haliotis tuberculata phenotypic traits

Abalone growth rate is often identified among important traits to improve through selective breeding. However, the rapid success of some selective breeding plans has sometimes led to negative effects in some aquaculture species due to trade-offs. One of them is the loss of homeostasis of selected animals which results in the inability to resist the stress experienced during the rearing process. In this context, this study aimed to analyze the phenotypic relationships between growth, and physiological and behavioural traits in Haliotis tuberculata under stressful conditions. Eleven traits related to growth, immunity, reproduction and behaviour were recorded under laboratory conditions. A total of 120 adults from wild or farm origin were first monitored during a 3-week stress period (high density and acute stress handling) during winter, followed by 6 months on-growing in sea-cages. Relationships between parameters were analyzed using a multi-factorial approach. Wild and farm stocks could only be discriminated on behaviour traits, confirming that the French abalone industry is in the beginning of the domestication process. After 3 weeks of chronic stress, the righting latency of an abalone was linked to better survival and faster growth. Abalones having the best growth after 6 months were characterized by higher activity during the previous winter period, whereas an early gonad maturation reduced the growth in summer. Our results provide a basis for the establishment of a multi-trait breeding program to improve the growth rate while controlling the evolution of physiological and behavioural traits. Statement of relevance The relationships of behavioural and physiological variables with survival and weight gain after application of important stressors were studied in order to provide a better understanding of Haliotis tuberculata biology during early domestication stage. This paper will give information on new targets and tools for selective breeding