Anne Cueff

Stress and stress-induced neuroendocrine changes increase the susceptibility of juvenile oysters (Crassostrea gigas) to Vibrio splendidus

ABSTRACT Oysters are permanently exposed to various microbes, and their defense system is continuously solicited to prevent accumulation of invading and pathogenic organisms. Therefore, impairment of the animals defense system usually results in mass mortalities in cultured oyster stocks or increased bacterial loads in food products intended for human consumption. In the present study, experiments were conducted to examine the effects of stress on the juvenile oysters resistance to the oyster pathogen Vibrio splendidus. Oysters (Crassostrea gigas) were challenged with a low dose of a pathogenic V. splendidus strain and subjected to a mechanical stress 3 days later. Both mortality andV.splendidus loads increased in stressed oysters, whereas they remained low in unstressed animals. Injection of noradrenaline or adrenocorticotropic hormone, two key components of the oyster neuroendocrine stress response system, also caused higher mortality and increased accumulation of V. splendidus in challenged oysters. These results suggest that the physiological changes imposed by stress, or stress hormones, influenced host-pathogen interactions in oysters and increased juvenile C. gigasvulnerability to Vibrio splendidus.

Stress-induced immune changes in the oyster Crassostrea gigas

Information concerning the effect of stress on invertebrate immune functions are scarce. The present study investigated the consequences of a 15-min mechanical disturbance on immune parameters in oysters Crassostrea gigas. As indicated by noradrenaline and dopamine measurements, the mechanical disturbance caused a transient state of stress in oysters. The number of circulating hemocytes, the migratory and phagocytic activities and reactive oxygen species production of hemocytes were measured before, during and after application of the stressor. Results show that all immune functions were significantly downregulated during stress and a transient period of immunostimulation was observed 30-240 min after the end of the disturbance. Taken together, these results suggest that stress can exert a profound influence on oyster immune functions and they may explain why stress and the outbreak of disease are often linked in shellfish culture. Furthermore, the present study strongly suggests that checking the stress status of animals may be necessary to avoid biases when studying oyster immune responses in vivo.

Cytotoxicity of diatom-derived oxylipins in organisms belonging to different phyla

SUMMARY The cytotoxicity of several saturated and unsaturated marine diatom-derived aldehydes and an oxo-acid have been screened in vitro and in vivo against different organisms, such as bacteria, algae, fungi, echinoderms, molluscs and crustaceans. Conjugated unsaturated aldehydes like 2E,4E-decadienal, 2E,4E-octadienal, 5E,7E-9-oxo-nonadienoic acid and 2E-decenal were active against bacteria and fungi and showed weak algicidal activity. By contrast, the saturated aldehyde decanal and the non-conjugated aldehyde 4Z-decenal had either low or no significant biological activity. In assays with oyster haemocytes, 2E,4E-decadienal exhibited a dose-dependent inhibition of cytoskeleton organisation, rate of phagocytosis and oxidative burst and a dose-dependent promotion of apoptosis. A maternal diatom diet that was rich in unsaturated aldehydes induced arrest of cell division and apoptotic cell degradation in copepod embryos and larvae, respectively. This wide spectrum of physiological pathologies reflects the potent cell toxicity of diatom-derived oxylipins, in relation to their non-specific chemical reactivity towards nucleophilic biomolecules. The cytotoxic activity is conserved across six phyla, from bacteria to crustaceans. Deregulation of cell homeostasis is supposed to induce the elimination of damaged cells through apoptosis. However, efficient protection mechanisms possibly exist in unicellular organisms. Experiments with a genetically modified yeast species exhibiting elevated membrane and/or cell wall permeability suggest that this protection can be related to the inability of the oxylipin compounds to enter the cell.

Noradrenaline modulates hemocyte reactive oxygen species production via β-adrenergic receptors in the oyster Crassostrea gigas

Catecholamines (CA) are known to be present in the microenvironment of molluscan immunocytes. In the present study, experiments were conducted to determine the effects of noradrenaline (NA), the principal CA circulating in bivalve hemolymph, on the luminol-dependent chemiluminescence (CL) of oyster Crassostrea gigas hemocytes. Results show that NA had a dose-dependent inhibitory effect on the CL-response at the physiological concentration of 0.1 microM and above. The alpha-adrenoceptor agonist phenylephrine had no significant effect on the CL-response whereas the beta-adrenoceptor agonist isoproterenol mimicked the inhibitory effects of NA on the CL-response. The beta-adrenoceptor antagonist propanolol, but not the alpha-adrenoceptor antagonist prazosin, prevented the negative effects of NA on the CL-response. Taken together, these results show that beta-adrenergic receptors are present at the surface of oyster hemocytes and allow NA to down-regulate the CL-response.

A first insight into stress-induced neuroendocrine and immune changes in the octopus Eledone cirrhosa

A number of cephalopod species present substantial ecological and economical importance; however, data on the physiology of stress and on regulatory processes linking stress to immune defence against pathogens remain extremely scarce in these organisms. The present study examined the influence of a 5 min air exposure, a common perturbation associated with handling in aquaculture settings and fisheries, on neuroendocrine and immune parameters in the octopus Eledone cirrhosa. Measurements of circulating concentrations of noradrenaline and dopamine, two hormones that are released in the haemolymph during stress in bivalves and gastropods, showed that the 5 min air exposure represents a real stress to octopus. Indeed, blood levels of both hormones increased by about 2‐2.5-fold in stressed animals. Concomitantly, a significant decrease in the number of circulating haemocytes was observed, whereas haemocyte phagocytotic activity and superoxide anion production increased transiently between 5 and 60 min after the beginning of the stress. These results provide a first insight into the effects of stress on catecholamine levels and immune functions in cephalopods and suggest that stress and immunity may be associated in these organisms.