Identification of apparently neurotoxic metabolites from assemblages of marine filamentous cyanobacteria associated with the intoxication of captive bottlenose dolphins (Tursiops truncatus) in the Florida Keys

Abstract

Intoxications of captive bottlenose dolphins (Tursiops truncatus) in the Florida Keys have been linked to observed interactions with marine macrophytic algal and cyanobacterial communities within enclosures. Taxonomic characterization of these communities coupled, in turn, to available observational data collected during intoxication events point to a contribution of filamentous cyanobacterial assemblages comprised of members of the polyphyletic genus, Lyngbya sensu lato. To identify toxic metabolites possibly relevant to these intoxications, chemical screening for known neurotoxins from cyanobacteria, as well as other regionally relevant harmful algal bloom (HAB) taxa, was combined with toxicity testing, and subsequent bioassay-guided fractionation, employing early life stages (i.e., embryos and larvae) of zebrafish (Danio rerio) as a well-established aquatic vertebrate toxicological model. Chemical analyses did not detect (within analytical limits) any of the known algal or cyanobacterial neurotoxins. Toxicity testing, alongside bioassay-guided fractionation, however, identified several chemical fractions with a range of potentially relevant bioactivities in both zebrafish embryos and post-hatch larvae including, in particular, behavioral (e.g., aberrant swimming) and physiological (e.g., altered heart rate) endpoints indicative of possible neurotoxicity, and subsequent chemical characterization of fractions suggested a contribution of the previously identified bioactive metabolite, eudesmacarbonate, in the observed toxicity. Comparative toxicological assessment with PbTx-2, as a positive control for neurotoxicity in the zebrafish model, further supported neurotoxic activity of cyanobacterial metabolites potentially relevant, in turn, to a contribution of these metabolites to dolphin intoxications. These findings suggest, in general, that marine zoological facilities may be affected by regional HABs, and assessments of potentially toxigenic algae and cyanobacteria should be included in management strategies in these facilities.

Keywords: Bottlenose dolphins; Brevetoxins; Harmful algal blooms; Lyngbya; Marine cyanotoxins; Neurotoxins.