Multiple toxic and bioactive compounds produced by Alexandrium spp. cause adverse effects on bivalves, but these effects are frequently difficult to attribute to a single compound class. To disentangle the effect of neurotoxic vs lytic secondary metabolites, we exposed blue mussels to either a paralytic shellfish toxin (PST) producing Alexandrium spp. strain, or to an exclusively lytic compound (LC) producing strain, or a strain containing both compound classes, to evaluate the time dependent effects after 3 and 7 days of feeding. Tested parameters comprised signs of paralysis, feeding activity, and immune cell integrity (hemocyte numbers and viability; lysosomal membrane destabilization) and function (ROS production). Both compound classes caused paralysis and immune impairment. The only effect attributable exclusively to PST was increased phagocytic activity after 3 days and impaired feeding activity after 7 days, which curtailed toxin accumulation in digestive glands. Lysosomal membrane destabilization were more closely, but not exclusively, matched with LC exposure. Effects on circulating hemocyte integrity and immune related functions were mostly transient or remained stable within 7 days; except for increased lysosomal labialization and decreased extracellular ROS production when mussels were exposed to the toxin combination. M. edulis displays adaptive fitness traits to survive and maintain immune capacity upon prolonged exposure to environmentally relevant concentrations of PST and/or LC producing Alexandrium strains.
We studied accumulation and biochemical effects of microcystin-LR (MCLR) in Odontesthes hatcheri after dietary administration of the cyanobacteria Microcystis aeruginosa (1.3 μg MCLR/g body mass, incorporated in standard fish food). After 12 h, MCLR content in liver did not differ between fish fed with crushed or intact cells, demonstrating O. hatcheri's capacity to digest cyanobacteria and absorb MCLR. In the second experiment, fish received toxic cells, non-toxic cells, or control food; MCLR accumulation was monitored for 48 h. Protein phosphatase 1 (PP1), catalase (CAT), glutathione-S-transferase (GST) activities, and lipid peroxidation (as MDA) were measured in liver and intestine. Methanol-extractable MCLR was determined by PP1 inhibition assay (PPIA); extractable and protein-bound MCLR were measured by Lemieux oxidation-gas chromatography/mass spectrometry (GC/MS). MCLR accumulated rapidly up to 22.9 and 9.4 μg MCLR/g in intestine and liver, respectively, followed by a decreasing tendency. Protein-bound MCLR represented 66 to ca. 100 % of total MCLR in both tissues. PP1 activity remained unchanged in intestine but was increased in liver of MCLR treated fish.CAT and GST activities and MDA content were significantly increased by MCLR only in liver. We conclude that O. hatcheri is able to digest cyanobacteria, accumulating MCLR mostly bound to proteins. Our data suggest that this freshwater fish can be adversely affected by cyanobacterial blooms. However, the rapid decrease of the detectable MCLR in both tissues could imply that sublethal toxin accumulation is rapidly reversed.
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