title: First report in a river in France of the benthic cyanobacterium Phormidium favosum producing anatoxin-a associated with dog neurotoxicosis.
abstract: The first identification of anatoxin-a in a French lotic system is reported. Rapid deaths of dogs occurred in 2003 after the animals drank water from the shoreline of the La Loue River in eastern France. Sediments, stones and macrophytes surfaces at the margin of the river were covered by a thick biofilm containing large quantities of several benthic species of filamentous, non-heterocystous cyanobacteria. Known cyanotoxins, such as microcystins, saxitoxins and anatoxins were screened from biofilm samples by biochemical and analytical assays. A compound with similar UV spectra to the anatoxin-a standard was detected by high-performance liquid chromatography (HPLC) coupled with photo-diode array detector. This toxin was further identified by HPLC coupled with a UV detector and by electrospray ionisation-Quadrupole-Time-Of-Flight mass spectrometer, and confirmed by tandem mass spectrometry. These two techniques were necessary to discriminate anatoxin-a in phenylalanine-containing matrices such as liver samples of poisoned dogs. The toxin and the aromatic amino acid, phenylalanine, present the same pseudomolecular ion at m/z 166, but have differing fragmentation patterns, retention times and UV spectra. Finally, several cyanobacterial strains were isolated from the green biofilm and tested for anatoxin-a production. Phormidium favosum was identified as a new anatoxin-a producing species.
authors.affiliation: USM0505 Ecosystèmes et interactions toxiques, M.N.H.N., Paris, France. mgugger@mnhn.fr
authors.forename: Muriel
authors.initials: M
authors.lastname: Gugger
authors.forename: Séverine
authors.initials: S
authors.lastname: Lenoir
authors.forename: Céline
authors.initials: C
authors.lastname: Berger
authors.forename: Aurélie
authors.initials: A
authors.lastname: Ledreux
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Druart
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Humbert
authors.forename: Catherine
authors.initials: C
authors.lastname: Guette
authors.forename: Cécile
authors.initials: C
authors.lastname: Bernard
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000032:analysis / Q000506:poisoning*; D045744:Cell Line, Tumor; D000458:Cyanobacteria / Q000737:chemistry / Q000145:classification* / Q000378:metabolism; D000087522:Cyanobacteria Toxins; D004285:Dogs; D004784:Environmental Monitoring; D005602:France; D005766:Gastrointestinal Contents / Q000737:chemistry; D007422:Intestines / Q000737:chemistry; D008099:Liver / Q000737:chemistry; D008387:Marine Toxins / Q000032:analysis / Q000506:poisoning*; D051379:Mice; D052998:Microcystins; D009498:Neurotoxins / Q000032:analysis / Q000506:poisoning*; D010456:Peptides, Cyclic / Q000032:analysis; D011041:Poisoning / Q000662:veterinary*; D045483:Rivers; D012530:Saxitoxin / Q000032:analysis; D014326:Tropanes
chemical_list: D001427:Bacterial Toxins; D000087522:Cyanobacteria Toxins; D008387:Marine Toxins; D052998:Microcystins; D009498:Neurotoxins; D010456:Peptides, Cyclic; D014326:Tropanes; D012530:Saxitoxin; C078588:microcystin; C509783:anatoxin a