BMAA extraction of cyanobacteria samples: which method to choose?

ß-N-Methylamino-L-alanine (BMAA), a neurotoxin reportedly produced by cyanobacteria, diatoms and dinoflagellates, is proposed to be linked to the development of neurological diseases. BMAA has been found in aquatic and terrestrial ecosystems worldwide, both in its phytoplankton producers and in several invertebrate and vertebrate organisms that bioaccumulate it. LC-MS/MS is the most frequently used analytical technique in BMAA research due to its high selectivity, though consensus is lacking as to the best extraction method to apply. This study accordingly surveys the efficiency of three extraction methods regularly used in BMAA research to extract BMAA from cyanobacteria samples. The results obtained provide insights into possible reasons for the BMAA concentration discrepancies in previous publications. In addition and according to the method validation guidelines for analysing cyanotoxins, the TCA protein precipitation method, followed by AQC derivatization and LC-MS/MS analysis, is now validated for extracting protein-bound (after protein hydrolysis) and free BMAA from cyanobacteria matrix. BMAA biological variability was also tested through the extraction of diatom and cyanobacteria species, revealing a high variance in BMAA levels (0.0080-2.5797 µg g(-1) DW).

Biotransfer of ß-N-methylamino-L-alanine (BMAA) in a eutrophicated freshwater lake.

ß-N-Methylamino-L-alanine (BMAA), a neurotoxic non-protein amino acid, plays a significant role as an environmental risk factor in neurodegenerative diseases, such as amyotrophic lateral sclerosis. BMAA producers occur globally, colonizing almost all habitats and represent species from distinct phytoplanktonic groups, i.e., cyanobacteria, diatoms, and dinoflagellates. Bioaccumulation of BMAA in invertebrate and vertebrate organisms has also been registered around the globe. In the Baltic Sea, BMAA has been detected in several commercial fish species, raising the question of the bioaccumulation of BMAA in Swedish limnic systems. Here we find the presence of BMAA in water samples from Lake Finjasjön and identify its bioaccumulation patterns in both plankti-benthivorous and piscivorous fish, according to fish species, total weight, gender, and season of collection. For the first time, a large number of fish individuals were used in order to draw conclusions on BMAA bioaccumulation in a closed ecological community based on a statistical approach. We may, therefore, conclude that feeding patterns (plankti-benthivorous) and increased age of fish may lead to a higher tissue concentration of BMAA.

An experimental study of toxin production in Arthrospira fusiformis (Cyanophyceae) isolated from African waters.

Arthrospira is one genus of cyanoprokaryota for which information on toxin production exists for only a few strains. The purpose of this study was to investigate whether strains of Arthrospira fusiformis produce intracellular toxic compounds such as microcystins and anatoxin-a. The study was based on three strains of Arthrospira, two strains isolated from wastewater ponds in Mozambique and one from Lake Nakuru, Kenya. These strains were cultivated experimentally in different light intensities and salinities. Microcystins were analysed by ELISA and HPLC and anatoxin-a by HPLC. Toxicity analysis of the three strains, following the growth cycle, detected neither microcystins nor anatoxin-a. The results indicated that the strains selected were not toxigenic under the experimental conditions applied. Thus, the strains of A. fusiformis tested in the present study could be considered candidates for use in different applications such as in food supplements and in aquaculture.