title: Accumulation of nodularin-like compounds from the cyanobacterium Nodularia spumigena and changes in acetylcholinesterase activity in the clam Macoma balthica during short-term laboratory exposure.
abstract: In this laboratory study the effects of the cyanobacterium Nodularia spumigena (strain AV1) that produces hepatotoxic nodularin (NODLN), non-toxic Nodularia sphaerocarpa (strain UP16f) and purified NODLN on the infaunal clam Macoma balthica from the Baltic Sea were examined. N. sphaerocarpa (2.4 and 12.5 mg dw l(-1)), N. spumigena (4 and 20 mg dw l(-1), intracellular NODLN content ca. 4 and 20 microg l(-1)) and purified NODLN (10 and 50 microg l(-1)) were applied in experimental tanks at 24 h intervals for 96 h. Water samples were taken during the experiment for the measurement of soluble NODLN concentrations. The concentrations of total hepatotoxins in the soft tissues were analysed with enzyme-linked immunosorbent assay (ELISA) and NODLN with high-performance liquid chromatography/diode array detection (HPLC/DAD). Acetylcholinesterase (AChE) activity was measured from the foot tissue samples taken at 0, 24 and 96 h. In the water phase, 60-100% of NODLN in the pure-toxin treatments and all the NODLN in N. spumigena treatments appeared as a yet unidentified form with NODLN-like spectral characteristics. The compound was present in similar quantities also in the non-toxic N. sphaerocarpa treatments. In the toxic N. spumigena treatments the tissue concentration of hepatotoxic NODLN-like compounds (measured with ELISA) increased from the control levels of 0.16 to 16.6 microg g(-1) dw (24 h), reaching 30.3 microg g(-1) dw at 96 h. However, <5% of the toxin detected by ELISA could be shown to be NODLN in HPLC/DAD analysis. Matrix-assisted laser desorption/ionisation-time of flight mass spectrometry (MALDI-TOF/MS) analyses revealed no NODLN-glutathione (GSH) conjugates in the tissues of M. balthica. Combining the responses in the AChE activity recorded after 24 and 96 h exposure, increases by 46% (N. spumigena) and 36% (soluble NODLN) compared with the control activity were observed in the low-level toxic exposures. Oppositely, decreases by 19% (N. spumigena) and 27% (soluble NODLN) of control activity were observed in the high-level exposures. Only the differences between the increased and decreased AChE activities were statistically significant, with individuals exposed to high levels expressing approximately 55% of the activity of those exposed to low concentrations. The results show that M. balthica readily ingests toxic N. spumigena and that accumulation of peptides takes place rapidly, which has potential food chain effects through toxin enrichment. However, it appears that M. balthica is at least partly able to metabolise NODLN. In addition to hepatotoxicity, NODLN seems to induce concentration-dependent neurotoxic effects; this must be taken into consideration when applying AChE activity as a biomarker of specific anthropogenic contamination (e.g. organophosphate and carbamate pesticides).
authors.affiliation: Finnish Institute of Marine Research, PO Box 33, FIN-00931 Helsinki, Finland. lehtonen@fimr.fi
authors.forename: Kari K
authors.initials: KK
authors.lastname: Lehtonen
authors.forename: Harri
authors.initials: H
authors.lastname: Kankaanpää
authors.forename: Sari
authors.initials: S
authors.lastname: Leiniö
authors.forename: Vesa O
authors.initials: VO
authors.lastname: Sipiä
authors.forename: Stephan
authors.initials: S
authors.lastname: Pflugmacher
authors.forename: Eva
authors.initials: E
authors.lastname: Sandberg-Kilpi
mesh_terms: D000110:Acetylcholinesterase / Q000187:drug effects*; D000704:Analysis of Variance; D000818:Animals; D001427:Bacterial Toxins / Q000493:pharmacokinetics / Q000633:toxicity; D049872:Bivalvia / Q000201:enzymology / Q000378:metabolism*; D002800:Cholinesterase Inhibitors / Q000493:pharmacokinetics / Q000633:toxicity; D000458:Cyanobacteria / Q000378:metabolism; D004784:Environmental Monitoring / Q000379:methods; D008099:Liver / Q000187:drug effects; D010456:Peptides, Cyclic / Q000493:pharmacokinetics* / Q000633:toxicity*; D023382:Toxicity Tests, Acute
chemical_list: D001427:Bacterial Toxins; D002800:Cholinesterase Inhibitors; D010456:Peptides, Cyclic; C063998:nodularin; D000110:Acetylcholinesterase