Next Article in Journal
Effects of Miniaturization of the Summer Phytoplankton Community on the Marine Ecosystem in the Northern East China Sea
Previous Article in Journal
Environmental Stressors and Pathology of Marine Molluscs
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:

No β-N-Methylamino-L-alanine (BMAA) Was Detected in Stranded Cetaceans from Galicia (North-West Spain)

Portuguese Institute of the Sea and Atmosphere (IPMA), Rua Alfredo Magalhães Ramalho, 6, 1495-006 Lisbon, Portugal
Centre of Marine Sciences (CCMAR/CIMAR LA), Campus de Gambelas, University of Algarve, 8005-139 Faro, Portugal
Department of Ecology, Environment and Plant Sciences, Stockholm University, 106 91 Stockholm, Sweden
Departamento de Biologia & CESAM, Campus Universitário de Santiago, Universidade de Aveiro, 3810-193 Aveiro, Portugal
Coordinadora para o Estudo dos Mamíferos Mariños (CEMMA), Rúa Cean 2, 36350 Nigrán, Spain
Author to whom correspondence should be addressed.
J. Mar. Sci. Eng. 2022, 10(3), 314;
Received: 25 December 2021 / Revised: 15 February 2022 / Accepted: 18 February 2022 / Published: 23 February 2022
(This article belongs to the Section Marine Biology)


The neurotoxin β-N-methylamino-L-alanine (BMAA), a non-proteinogenic amino acid produced by several species of both prokaryotic (cyanobacteria) and eukaryotic (diatoms) microorganisms, has been proposed to be associated with the development of neurodegenerative diseases. At first, BMAA appeared to be ubiquitously present worldwide in various organisms, from aquatic and terrestrial food webs. However, recent studies, using detection methods based on mass spectrometry, instead of fluorescence detection, suggest that the trophic transfer of BMAA is debatable. This study evaluated BMAA in 22 cetaceans of three different species (Phocoena phocoena, n = 8, Delphinus delphis, n = 8, and Tursiops truncatus, n = 6), found stranded in North-West Spain. BMAA analysis of the liver, kidney, or muscle tissues via sensitive liquid chromatography with tandem mass spectrometry did not reveal the presence of this compound or its isomers. The absence recorded in this study highlights the need to better understand the trophic transfer of BMAA and its anatomical distribution in marine mammals.

1. Introduction

The non-proteinogenic neurotoxin β-N-methylamino-L-alanine (BMAA) has been proposed to act as an environmental factor, inducing the development of several neurodegenerative diseases, such as amyotrophic lateral sclerosis (ALS), Alzheimer’s disease, and Parkinson’s disease [1,2,3,4]. However, the hypothesis of a causative association between dietary exposure to BMAA and a neurodegenerative pathological condition remains controversial [5,6,7,8]. Cox et al. (2005) [9] detected BMAA in 95% of all cyanobacterial genera tested and concluded that BMAA was universally produced by cyanobacteria species from aquatic and terrestrial habitats, in both symbiotic and free-living forms. However, this conclusion was questioned, due to the non-specificity of the analytical method used. The positive detection of BMAA, with liquid chromatography (LC) or gas chromatography (GC), associated with ultraviolet, fluorescence spectroscopy, or single mass spectrometry (MS), is just based on the retention time and signal of the parent ion [10,11,12]. All these analytical methods might give false-positive results, considering that BMAA might co-elute with its natural isomers (i.e., DAB, 2,4-diaminobutyric acid; BAMA, β-amino-N-methyl-alanine; AEG, N-2(aminoethyl)glycine; DABA, 2,3-diaminobutyric acid; 3,4-diaminobutyric acid; 3-amino-2-(aminomethyl)-propanoic acid; and 2,3-diamino-2-methylpropanoic acid) or other interfering compounds [13]. Currently, it is accepted that only the use of liquid chromatography with tandem mass spectrometry detection (LC-MS/MS), with or without previous derivatization, ensures a reliable BMAA identification, based on the retention time, mass-to-charge ratio (m/z) of the precursor ion, and product fragmentions after collision-induced dissociation, and the ratio between the intensities of respective ions transitions in multiple reaction monitoring (MRM) spectrum [10,13]. Moreover, recent studies, using LC-MS/MS, have shown that most cyanobacteria produce only trace levels of BMAA [14,15].
In addition to cyanobacteria, several diatoms species produce BMAA and its structural isomers [16,17,18]. Several of these potential BMAA-producers are present in our study area, the coast of Galicia in North-West (NW) Spain (Figure 1), i.e., the diatoms Chaetoceros spp., Navicula spp., Skeletonema spp., and Thalassiosira spp. [19,20,21,22,23]. and cyanobacteria from the genera Anabaena, Myxosarcina, Lyngbya, Phormidium, Symploca, Nodularia, Nostoc, Calothrix, and Microcystis [20]. Moreover, the predominant wind and oceanographic conditions of NW Spain enhance phytoplankton production, and diatoms and dinoflagellates blooms are recurrently reported [20].
The presence of BMAA has been described in several organisms along the aquatic and terrestrial food chains, including zooplankton [24,25,26], crustaceans [27,28], bivalves [25,29,30,31], fish [25,32], and terrestrial plants and animals [33,34,35,36]. However, a review by Lance et al. (2018) [37] observed no clear indication of the BMAA trophic transfer. If BMAA is bioaccumulated through the marine food web, it is plausible that the highest concentration of BMAA would be present in apex predators [38,39,40]. Massive strands of marine mammals, due to harmful algal events, have been reported worldwide, and evidences of their recurrent exposure to these toxins was also suggested [41,42,43,44]. Cetaceans are important key species for marine ecosystems, being excellent indicators of environmental changes. Indeed, recent studies performed in the West Atlantic confirmed that common and bottlenose dolphins are susceptible to BMAA accumulation and damage [8,45].
Galicia, located in NW Spain (Figure 1), holds important populations of marine mammals, which reckon 23 species [46,47]. Among them, the common dolphin (Delphinus delphis), bottlenose dolphin(Tursiops truncatus), and harbour porpoise (Phocoena phocoena) are the most frequently found washed ashore [48,49,50]. The harbour porpoise in the area has been identified as a new ecotype and proposed as a new subspecies [51].
Therefore, to explore the potential BMAA trophic transfer, and its potential association with the stranding of cetaceans, we analysed the BMAA, DAB, and AEG levels in three different tissues (liver, kidney, and muscle) of 22 individuals from three cetacean species, namely the common dolphin, bottlenose dolphin, and harbour porpoise. All individuals were found stranded between 2011 and 2017.

2. Materials and Methods

A stranding network was established in Galicia, in 1990, and carried out by NGO Coordinadora para o Estudo dos MamíferosMariños (CEMMA), to locate beached cetaceans, pinnipeds, and sea turtles, ensuring the biological samples collection and rehabilitation actions for live animals. Basic data recorded includes species, biometrics, gender identification, body condition, external examination for each animal, and signs of bycatch. Any other relevant details were also recorded (Table 1). Necropsies were carried out on fresh and moderately decomposed animals. Both external studies and necropsies followed standardized protocols [52,53,54]. Figure 1 displays the location where the animals, used in this study, were found.
Tissue samples of liver, kidney, and muscle were frozen at −20 °C for preservation. Before analyses, the samples were freeze-dried (Freeze Dry System Labconco, Coolvacuum Technologies, Barcelona, Spain). The BMAA (total soluble and precipitated bound) was extracted, based on Murch et al. (2004) [55], with minor alterations, suggested byMasseret et al. (2013) [56] and Lage et al. (2016) [57], as previously described [29,57,58]. The tissue samples (2 mg dry weight each) were extracted in triplicate. After extraction, freeze-dried BMAA total soluble and precipitated bound samples were reconstituted with 20 mM HCl solution and dilutions were performed (if required), to obtain an optimum ratio of protein-to-derivatization agent ratio [57]. Subsequently, the samples were derivatized with AccQ-Tag, using a WAT052880 AccQ-Tag kit (Waters, Milford, MA, USA) before analysis.
Samples of each tissue type (liver, kidney, and muscle) and BMAA form (i.e., total soluble and precipitated bound)were used for the analysis of limits of quantification; LOQ was defined as S/N ≥ 10 [59]. Samples of liver BMAA precipitated bound form were used for the evaluation of the matrix effect and extraction method recovery at the BMAA concentrations of 5 and 10 ng mL−1 (n = 4).
LC-MS/MS analysis of derivatized BMAA, and its isomers AEG and DAB, were performed in an Acquity UPLC system, coupled witha Xevo-TQ-MS system (Waters, Milford, MA, USA), as previously described [30].The LC separation was performed on an AccQ-Tag Ultra C18 column (100 × 2.1 mm, 1.7 μm particle size, Waters, Milford, MA, USA). Ionization was performed in positive ion mode, and the mass analyser was run in the selected reaction monitoring (SRM) scanning mode, using the following transitions to distinguish BMAA from its isomers, AEG and DAB: common to all three analytes, 459.1 > 119.1 (CE 30.0); DAB diagnostic fragment, 459.1 > 188.1(CE 38.0); BMAA diagnostic fragment, 459.1 > 258.1 (CE 30.0); and AEG diagnostic fragment, 459.1 > 214.1 (CE 30.0). To ensure the accurate identification of BMAA, the parameters retention time and fragmentation ratio of the fragments, 119.1/258.1, were considered. All settings were optimized for the detection of BMAA, as follows: spray voltage, 5500 V; source temperature, 450 °C; decluttering potential, 50; focusing potential, 350; and entrance potential, 6.MassLynx V4.1 software (Waters, Milford, MA, USA) was used to analyse the acquired data.

3. Results and Discussion

No measurable levels of BMAA and its structural isomers (AEG and DAB) were detected, in either the total soluble or precipitated bound forms of the three tissues (liver, kidney, and muscle) of the 22 specimens studied. To the best of our knowledge, this is the first study analysing the liver, kidney, and muscle tissue samples of cetaceans for the presence of BMAA and its structural isomers. Previously, Davis et al. (2019) reported total BMAA concentrations, ranging from 20 to 748 μg−1, in the brains of 13 dolphins, found stranded in Florida and Massachusetts, USA. Furthermore, dolphins with detectable levels of BMAA presented injuries in the cerebral cortex and increased β-amyloid plaques [8]. Unfortunately, the discrepancy in dolphins’ tissues, analysed by Davis et al. (2019) (i.e., brain) and the current study (i.e., liver, kidney, and muscle), does not allow a direct comparison.
Although the brains of the animals were not available for the present study, considering the lack of data on BMAA accumulation in marine mammals, we judged that the analyses of other tissues may still be of relevance. In fish species, the four tissues (brain, liver, kidney, and muscle) have been previously analysed, thus providing an estimation of the BMAA anatomical distribution [25,60]. In several fish species, collected in the Baltic Sea and in Lake Finjasjön (Sweden), the highest BMAA levels were found in the fish brains [25,60]. The total BMAA concentrations in the fish brains were up to 0.99 and 0.028 µg g−1 DW, while in the fish muscle were up to 0.059 and 0.006 µg g−1 DW in the Baltic Sea and Lake Finjasjön, respectively [25,60]. Moreover, from the total of 136 fish individuals analysed by Lage et al. (2015), only 22 individuals (16%) contained quantifiable BMAA in their muscle, while BMAA was quantified in the brains of 40 individuals (29%) [60]. No BMAA was detected in the kidney and liver of fish collected in the Baltic Sea and Lake Finjasjön [25,60]. In other studies of fish collected in the Baltic Sea, the Eastern North Atlantic, and the Mediterranean Sea, no BMAA was detected in the fish muscle [16,26,28,61].
Tissue composition may play an important role in BMAA accumulation. BMAA is misincorporated, instead of L-serine during protein synthesis. Moreover, BMAA anatomical distribution in adult mice showed a distribution pattern analogous to protein-forming amino acids [62,63]. Another study in neonatal rats reported a higher uptake and retention of BMAA in tissues, with high rates of protein synthesis and cell turnover, suggesting that BMAA may be incorporated or associated with newly synthesized proteins [64]. However, BMAA was cleared out of the body over time. Thus, the non-detection of BMAA in the muscle, liver, and kidney of cetaceans found stranded in Galicia might be due to the higher turn-over rate of these tissues, leading to the degradation and release of BMAA, especially if the cetaceans were starved for a long period of time.
Although brain samples were not analysed, the muscle of several fish species caught in Lake Taihu (China) had total BMAA concentrations higher than the muscle of fish caught in the Baltic Sea, Eastern North Atlantic, and Mediterranean Sea, with concentrations ranging from 0.07 and 35.91 µg g−1 DW [65]. Moreover, sharks, apex predators caught in South Florida (USA), had total BMAA concentrations between 19.2 and 33.15 µg g−1 FW in the fins and muscle [32,66]. Furthermore, dietary supplements containing shark cartilage, from various species and origins (not reported), had total BMAA concentrations between 74.8 and 352.2 µg g−1 DW [67]. BMAA contents in organisms may vary, depending on the methodological differences between studies, inter-specific variations in their trophic status, geographical and seasonal parameters, and ecological responses of its producers [37]. Thus, as previously documented for fish [16,25,26,28,32,60,61], dolphins of certain geographical areas might also contain higher concentrations of BMAA than others. Accordingly, Davis et al. (2019) reported three-fold higher concentrations of total BMAA in dolphins stranded in Florida than in dolphins from Massachusetts [45].
Methodological differences between studies are often responsible for the variability of data reported on BMAA concentration in biota, especially when dealing with complex tissues, which may induce matrix interferences [10,37,68]. A previously published and in-house validated LC-MS/MS method [56,57,69], which had minor differences from the method used by Davis et al. (2019,2021), was used in the present study [8,45]. The LOQ on BMAA spiked tissue samples was 0.5 ng mL−1 (corresponding to 0.11 μg g−1), and the matrix effect in the liver tissue precipitated bound BMAA samples was 64.04 ± 3.77 and 54.22 ±6.14% in the 5 and 10 ng mL−1 spiked concentrations, respectively (Figure 2). The BMAA recovery rates in the 5 and 10 ng mL−1 spiked BMAA liver samples were 96.62 ± 20.12 and 99.20 ± 17.44%, respectively. Davis et al. (2019, 2021) reported a LOQ of 7.0 ng mL−1 and an average % recovery of BMAA of 98.3% [8,45]. Moreover, matrix spiked recovery (%) of several metabolites has been previously shown to be similar among various cetaceans tissues, i.e., blubber, muscle, liver, kidney, stomach, melon, and gonad [70]. Therefore, if the tissues analysed in the present study had BMAA concentrations comparable with the concentrations reported for the brains of cetaceans stranded in the USA, they would have been quantified. Unfortunately, as we did not analyse brain tissues or cerebral spinal fluid, we cannot rule out higher toxin levels in these tissues.
The absence of BMAA in our samples is unexpected. In Galicia, the phytoplankton successions, during the upwelling system dynamics, are characterized by the dominance of diatoms from late-winter to summer [19,20]. Among these diatoms, several potential BMAA-producers are accounted reaching high densities [19,20,21,22,23]. The estuaries (“rías”) receive nutrients and phytoplankton from rivers, and high abundances of cyanobacteria, including potential BMAA-producers, are frequent in moderately stratified waters [71,72,73,74,75]. Unfortunately, BMAA levels for these bloom periods are unknown. Regulated toxins, such as amnesic toxins (domoic acid), paralytic shellfish toxins (saxitoxins), and the diarrhetic shellfish toxins (okadaic acid and derivatives), are regularly monitored, according to EU directives, to ensure human food safety [76]. However, BMAA lacks specific regulations and monitoring plans.
The BMAA absence, recorded in this study, highlights the need to better understand the trophic transfer of BMAA, depending on environment conditions and its anatomical distribution in marine mammals.

4. Conclusions

Neither the biotoxin BMAA nor their isomers were detected in several specimens of dolphins washed ashore in the Galicia coastline, Spain. Top predators, such as cetaceans, may reflect the overall BMAA incidence in the marine environment, which makes them suitable sentinel species for biotoxin exposure risk. The absence of BMAA in the liver, kidney, and muscle of the 22 individuals analysed might indicate a lower incidence of BMAA producers in this geographical area or lack of BMAA trophic transfer, but research on differential toxin accumulation in tissues and organisms needs further attention.

Author Contributions

Conceptualization, S.L., L.S. and P.R.C.; methodology, S.L. and S.-Y.K.; validation, S.L. and S.-Y.K.; formal analysis, S.L.; investigation, L.S., S.L., S.-Y.K., A.L. and P.C.; resources, S.R.; writing—original draft preparation, L.S. and S.L.; writing—review and editing, all authors; supervision, S.L., S.R. and P.R.C. All authors have read and agreed to the published version of the manuscript.


L.S. was supported by the project Cigua (PTDC/CTA-AMB/30557/2017), funded by the Portuguese Foundation for Science and Technology (FCT) and FEDER (MAR2020). This study also received Portuguese national funds from FCT, through project UIDB/Multi/04326/2020. Funding for sample collection was provided for Galicia by Direccion Xeral de Patrimonio Natural-CMA-Xunta de Galicia. The work of A.L. is supported by the CESAM by FCT/MCTES (UIDP/50017/2020+UIDB/50017/2020+LA/P/0094/2020), through national funds and by national funds (OE), through FCT-I.P., in the scope of the framework contract foreseen in the numbers 4, 5 and 6 of the article 23, of the Decree-Law 57/2016, of August 29, changed by Law 57/2017, of July 19. SY.K. and S.R. were supported by funding from the Swedish Research Council Formas. The foundation BalticSea2020 and Stiftelsen Olle Engkvist Byggmästare funded the Acquity UPLC couple to Xevo-TQ-MS (Waters) via the MiMeBS program grant, awarded to Professor Birgitta Bergman. S.L. was funded by H2020-WF-02-2019, grant no.101003376.

Institutional Review Board Statement

Legal permits for sample collection was provided for Galicia by Direccion Xeral de Patrimonio Natural-CMA-Xunta de Galicia, and samples were provided by the “Coordinadora para o Estudo dos MamíferosMariños” (CEMMA).

Informed Consent Statement

Not applicable.

Conflicts of Interest

The authors declare no conflict of interest.


  1. Nunn, P.B. 50 years of research on α-amino-β-methylaminopropionic acid (β-methylaminoalanine). Phytochemistry 2017, 144, 271–281. [Google Scholar] [CrossRef]
  2. Spencer, P.S.; Nunn, P.B.; Hugon, J.; Ludolph, A.C.; Ross, S.M.; Roy, D.N.; Robertson, R.C. Guam Amyotrophic Lateral Sclerosis-Parkinsonism-Dementia linked to a plant excitant neurotoxin. Science 1987, 237, 517–522. [Google Scholar] [CrossRef]
  3. Cox, P.A.; Sacks, O.W. Cycad neurotoxins, consumption of flying foxes, and ALS-PDC disease in Guam. Neurology 2002, 58, 956–959. [Google Scholar] [CrossRef]
  4. Bradley, W.G.; Mash, D.C. Beyond Guam: The cyanobacteria/BMAA hypothesis of the cause of ALS and other neurodegenerative diseases. Amyotroph. Lateral Scler. 2009, 10, 7–20. [Google Scholar] [CrossRef]
  5. Cox, P.A.; Davis, D.A.; Mash, D.C.; Metcalf, J.S.; Banack, S.A. Dietary exposure to an environmental toxin triggers neurofibrillary tangles and amyloid deposits in the brain. Proc. R. Soc. B Biol. Sci. 2016, 283, 20152397. [Google Scholar] [CrossRef][Green Version]
  6. Cox, P.A.; Davis, D.A.; Mash, D.C.; Metcalf, J.S.; Banack, S.A. Do vervets and macaques respond differently to BMAA? Neurotoxicology 2016, 57, 310–311. [Google Scholar] [CrossRef]
  7. Lobner, D.; Piana, P.M.T.; Salous, A.K.; Peoples, R.W. β-N-methylamino-l-alanine enhances neurotoxicity through multiple mechanisms. Neurobiol. Dis. 2007, 25, 360–366. [Google Scholar] [CrossRef][Green Version]
  8. Davis, D.A.; Garamszegi, S.P.; Banack, S.A.; Dooley, P.D.; Coyne, T.M.; McLean, D.W.; Rotstein, D.S.; Mash, D.C.; Cox, P.A. BMAA, Methylmercury, and mechanisms of neurodegeneration in dolphins: A natural model of toxin exposure. Toxins 2021, 13, 697. [Google Scholar] [CrossRef]
  9. Cox, P.A.; Banack, S.A.; Murch, S.J.; Rasmussen, U.; Tien, G.; Bidigare, R.R.; Metcalf, J.S.; Morrison, L.F.; Codd, G.A.; Bergman, B. Diverse taxa of cyanobacteria produce β-N-methylamino-L-alanine, a neurotoxic amino acid. Proc. Natl. Acad. Sci. USA 2005, 102, 5074–5078. [Google Scholar] [CrossRef][Green Version]
  10. Faassen, E.J.; Gillissen, F.; Lürling, M. A comparative study on three analytical methods for the determination of the neurotoxin BMAA in cyanobacteria. PLoS ONE 2012, 7, e36667. [Google Scholar] [CrossRef]
  11. Faassen, E.J. Presence of the neurotoxin BMAA in aquatic ecosystems: What do we really know? Toxins 2014, 6, 1109–1138. [Google Scholar] [CrossRef][Green Version]
  12. Krüger, T.; Mönch, B.; Oppenhäuser, S.; Luckas, B. LC–MS/MS determination of the isomeric neurotoxins BMAA (β-N-methylamino-l-alanine) and DAB (2,4-diaminobutyric acid) in cyanobacteria and seeds of Cycas revoluta and Lathyrus latifolius. Toxicon 2010, 55, 547–557. [Google Scholar] [CrossRef]
  13. Jiang, L.; Aigret, B.; De Borggraeve, W.M.; Spacil, Z.; Ilag, L.L. Selective LC-MS/MS method for the identification of BMAA from its isomers in biological samples. Anal. Bioanal. Chem. 2012, 403, 1719–1730. [Google Scholar] [CrossRef]
  14. Chernoff, N.; Hill, D.J.; Diggs, D.L.; Faison, B.D.; Francis, B.M.; Lang, J.R.; Larue, M.M.; Le, T.-T.; Loftin, K.A.; Lugo, J.N.; et al. A critical review of the postulated role of the non-essential amino acid, β-N-methylamino-L-alanine, in neurodegenerative disease in humans. J. Toxicol. Environ. Health Part B 2017, 20, 183–229. [Google Scholar] [CrossRef]
  15. Dunlop, R.A.; Banack, S.A.; Bishop, S.; Metcalf, J.S.; Murch, S.J.; Davis, D.A.; Stommel, E.W.; Karlsson, O.; Brittebo, E.B.; Chatziefthimiou, A.D.; et al. Is exposure to BMAA a risk factor for neurodegenerative diseases? A response to a critical review of the BMAA hypothesis. Neurotox. Res. 2021, 39, 81–106. [Google Scholar] [CrossRef]
  16. Jiang, L.; Eriksson, J.; Lage, S.; Jonasson, S.; Shams, S.; Mehine, M.; Ilag, L.L.; Rasmussen, U. Diatoms: A novel source for the neurotoxin BMAA in aquatic environments. PLoS ONE 2014, 9, e84578. [Google Scholar] [CrossRef]
  17. Wang, C.; Yan, C.; Qiu, J.; Liu, C.; Yan, Y.; Ji, Y.; Wang, G.; Chen, H.; Li, Y.; Li, A. Food web biomagnification of the neurotoxin β-N-methylamino-L-alanine in a diatom-dominated marine ecosystem in China. J. Hazard. Mater. 2021, 404, 124217. [Google Scholar] [CrossRef]
  18. Violi, J.P.; Facey, J.A.; Mitrovic, S.M.; Colville, A.; Rodgers, K.J. Production of β-methylamino-L-alanine (BMAA) and its isomers by freshwater diatoms. Toxins 2019, 11, 512. [Google Scholar] [CrossRef][Green Version]
  19. Prego, R.; Guzmán-Zuñiga, D.; Varela, M.; de Castro, M.; Gómez-Gesteira, M. Consequences of winter upwelling events on biogeochemical and phytoplankton patterns in a western Galician ria (NW Iberian peninsula). Estuar. Coast. Shelf Sci. 2007, 73, 409–422. [Google Scholar] [CrossRef]
  20. Estrada, M. Phytoplankton distribution and composition off the coast of Galicia (northwest of Spain). J. Plankton Res. 1984, 6, 417–434. [Google Scholar] [CrossRef]
  21. Varela, M.; Prego, R.; Pazos, Y.; Moroño, Á. Influence of upwelling and river runoff interaction on phytoplankton assemblages in a Middle Galician Ria and comparison with northern and southern rias (NW Iberian Peninsula). Estuar. Coast. Shelf Sci. 2005, 64, 721–737. [Google Scholar] [CrossRef]
  22. Réveillon, D.; Séchet, V.; Hess, P.; Amzil, Z. Production of BMAA and DAB by diatoms (Phaeodactylum tricornutum, Chaetoceros sp., Chaetoceros calcitrans and, Thalassiosira pseudonana) and bacteria isolated from a diatom culture. Harmful Algae 2016, 58, 45–50. [Google Scholar] [CrossRef]
  23. Tilstone, G.H.; Miguez, B.M.; Figueiras, F.G. Diatom dynamics in a coastal ecosystem affected by upwelling: Coupling between species succession, circulation and biogeochemical processes. Mar. Ecol. Prog. Ser. 2000, 205, 23–41. [Google Scholar] [CrossRef][Green Version]
  24. Christensen, S.J.; Hemscheidt, T.K.; Trapido-Rosenthal, H.; Laws, E.A.; Bidigare, R.R. Detection and quantification of β-methylamino-L-alanine in aquatic invertebrates. Limnol. Oceanogr. Methods 2012, 10, 891–898. [Google Scholar] [CrossRef]
  25. Jonasson, S.; Eriksson, J.; Berntzon, L.; Spáčil, Z.; Ilag, L.L.; Ronnevi, L.-O.; Rasmussen, U.; Bergman, B. Transfer of a cyanobacterial neurotoxin within a temperate aquatic ecosystem suggests pathways for human exposure. Proc. Natl. Acad. Sci. USA 2010, 107, 9252–9257. [Google Scholar] [CrossRef][Green Version]
  26. Zguna, N.; Karlson, A.M.L.; Ilag, L.L.; Garbaras, A.; Gorokhova, E. Insufficient evidence for BMAA transfer in the pelagic and benthic food webs in the Baltic Sea. Sci. Rep. 2019, 9, 10406. [Google Scholar] [CrossRef][Green Version]
  27. Jiang, L.; Kiselova, N.; Rosén, J.; Ilag, L.L. Quantification of neurotoxin BMAA (β-N-methylamino-L-alanine) in seafood from Swedish markets. Sci. Rep. 2014, 4, 6931. [Google Scholar] [CrossRef][Green Version]
  28. Salomonsson, M.L.; Fredriksson, E.; Alfjorden, A.; Hedeland, M.; Bondesson, U. Seafood sold in Sweden contains BMAA: A study of free and total concentrations with UHPLC–MS/MS and dansyl chloride derivatization. Toxicol. Rep. 2015, 2, 1473–1481. [Google Scholar] [CrossRef][Green Version]
  29. Braga, A.C.; Lage, S.; Pacheco, M.; Rydberg, S.; Costa, P.R. Native (Ruditapes decussatus) and non-indigenous (R. philippinarum) shellfish species living in sympatry: Comparison of regulated and non-regulated biotoxins accumulation. Mar. Environ. Res. 2017, 129, 147–155. [Google Scholar] [CrossRef]
  30. Lage, S.; Costa, P.R.; Moita, T.; Eriksson, J.; Rasmussen, U.; Rydberg, S.J. BMAA in shellfish from two Portuguese transitional water bodies suggests the marine dinoflagellate Gymnodinium catenatum as a potential BMAA source. Aquat. Toxicol. 2014, 152, 131–138. [Google Scholar] [CrossRef]
  31. Réveillon, D.; Séchet, V.; Hess, P.; Amzil, Z. Systematic detection of BMAA (β-N-methylamino-l-alanine) and DAB (2,4-diaminobutyric acid) in mollusks collected in shellfish production areas along the French coasts. Toxicon 2016, 110, 35–46. [Google Scholar] [CrossRef] [PubMed][Green Version]
  32. Mondo, K.; Hammerschlag, N.; Basile, M.; Pablo, J.; Banack, S.A.; Mash, D.C. Cyanobacterial neurotoxin β-N-methylamino-L-alanine (BMAA) in shark fins. Mar. Drugs 2012, 10, 509–520. [Google Scholar] [CrossRef] [PubMed][Green Version]
  33. Andersson, M.; Karlsson, O.; Brandt, I. The environmental neurotoxin β-N-methylamino-l-alanine (l-BMAA) is deposited into birds’ eggs. Ecotoxicol. Environ. Saf. 2018, 147, 720–724. [Google Scholar] [CrossRef] [PubMed]
  34. Field, N.C.; Metcalf, J.S.; Caller, T.A.; Banack, S.A.; Cox, P.A.; Stommel, E.W. Linking β-methylamino-l-alanine exposure to sporadic amyotrophic lateral sclerosis in Annapolis, MD. Toxicon 2013, 70, 179–183. [Google Scholar] [CrossRef]
  35. Li, B.; Yu, S.; Li, G.; Chen, X.; Huang, M.; Liao, X.; Li, H.; Hu, F.; Wu, J. Transfer of a cyanobacterial neurotoxin, β-methylamino-l-alanine from soil to crop and its bioaccumulation in Chinese cabbage. Chemosphere 2019, 219, 997–1001. [Google Scholar] [CrossRef]
  36. Metcalf, J.S.; Banack, S.A.; Kotut, K.; Krienitz, L.; Codd, G.A. Amino acid neurotoxins in feathers of the Lesser Flamingo, Phoeniconaias minor. Chemosphere 2013, 90, 835–839. [Google Scholar] [CrossRef]
  37. Lance, E.; Arnich, N.; Maignien, T.; Biré, R. Occurrence of β-N-methylamino-l-alanine (BMAA) and isomers in aquatic environments and aquatic food sources for humans. Toxins 2018, 10, 83. [Google Scholar] [CrossRef][Green Version]
  38. Dunlop, R.A.; Cox, P.A.; Banack, S.A.; Rodgers, K.J. The non-protein amino acid BMAA is misincorporated into human proteins in place of l-Serine causing protein misfolding and aggregation. PLoS ONE 2013, 8, e75376. [Google Scholar] [CrossRef][Green Version]
  39. Xie, X.; Basile, M.; Mash, D.C. Cerebral uptake and protein incorporation of cyanobacterial toxin β-N-methylamino-L-alanine. Neuroreport 2013, 24, 779–784. [Google Scholar] [CrossRef]
  40. Glover, W.B.; Mash, D.C.; Murch, S.J. The natural non-protein amino acid N-β-methylamino-l-alanine (BMAA) is incorporated into protein during synthesis. Amino Acids 2014, 46, 2553–2559. [Google Scholar] [CrossRef]
  41. Lefebvre, K.A.; Powell, C.L.; Busman, M.; Doucette, G.J.; Moeller, P.D.R.; Silver, J.B.; Miller, P.E.; Hughes, M.P.; Singaram, S.; Silver, M.W.; et al. Detection of domoic acid in northern anchovies and California sea lions associated with an unusual mortality event. Nat. Toxins 1999, 7, 85–92. [Google Scholar] [CrossRef]
  42. Scholin, C.A.; Gulland, F.; Doucette, G.J.; Benson, S.; Busman, M.; Chavez, F.P.; Cordaro, J.; DeLong, R.; De Vogelaere, A.; Harvey, J.; et al. Mortality of sea lions along the central California coast linked to a toxic diatom bloom. Nature 2000, 403, 80–84. [Google Scholar] [CrossRef]
  43. Lefebvre, K.A.; Bargu, S.; Kieckhefer, T.; Silver, M.W. From sanddabs to blue whales: The pervasiveness of domoic acid. Toxicon 2002, 40, 971–977. [Google Scholar] [CrossRef]
  44. Lefebvre, K.A.; Quakenbush, L.; Frame, E.; Huntington, K.B.; Sheffield, G.; Stimmelmayr, R.; Bryan, A.; Kendrick, P.; Ziel, H.; Goldstein, T.; et al. Prevalence of algal toxins in Alaskan marine mammals foraging in a changing arctic and subarctic environment. Harmful Algae 2016, 55, 13–24. [Google Scholar] [CrossRef][Green Version]
  45. Davis, D.A.; Mondo, K.; Stern, E.; Annor, A.K.; Murch, S.J.; Coyne, T.M.; Brand, L.E.; Niemeyer, M.E.; Sharp, S.; Bradley, W.G.; et al. Cyanobacterial neurotoxin BMAA and brain pathology in stranded dolphins. PLoS ONE 2019, 14, e0213346. [Google Scholar] [CrossRef] [PubMed]
  46. López, A. Mammalia. In Inventario de la Biodiversidad Marina de Galicia: Proyecto LEMGAL; Bañón, R., Ed.; Consellería do Mar, Xunta de Galicia, Santiago de Compostela: Santiago de Compostela, Spain, 2017; pp. 567–570. [Google Scholar]
  47. Covelo, P.; López, A. First record of dwarf sperm whale (Kogia sima) in the north of Spain. Galemys Span. J. Mammal. 2021, 33, 64–69. [Google Scholar] [CrossRef]
  48. López, A.; Pierce, G.J.; Valeiras, X.; Santos, M.B.; Guerra, A. Distribution patterns of small cetaceans in Galician waters. J. Mar. Biol. Assoc. UK 2004, 84, 283–294. [Google Scholar] [CrossRef][Green Version]
  49. Pierce, G.J.; Caldas, M.; Cedeira, J.; Santos, M.B.; Llavona, Á.; Covelo, P.; Martinez, G.; Torres, J.; Sacau, M.; López, A. Trends in cetacean sightings along the Galician coast, north-west Spain, 2003–2007, and inferences about cetacean habitat preferences. J. Mar. Biol. Assoc. UK 2010, 90, 1547–1560. [Google Scholar] [CrossRef]
  50. López, A.; Santos, M.B.; Pierce, G.J.; González, A.F.; Valeiras, X.; Guerra, A. Trends in strandings and by-catch of marine mammals in north-west Spain during the 1990s. J. Mar. Biol. Assoc. UK 2002, 82, 513–521. [Google Scholar] [CrossRef][Green Version]
  51. Fontaine, M.C.; Roland, K.; Calves, I.; Austerlitz, F.; Palstra, F.P.; Tolley, K.A.; Ryan, S.; Ferreira, M.; Jauniaux, T.; Llavona, A.; et al. Postglacial climate changes and rise of three ecotypes of harbour porpoises, Phocoena phocoena, in western Palearctic waters. Mol. Ecol. 2014, 23, 3306–3321. [Google Scholar] [CrossRef]
  52. Kuiken, T.; Hartmann, M.G. European Cetacean Society: Pacific Grove. In Proceedings of the First ECS Workshop on Cetacean Pathology: Dissection Techniques and Tissue Sampling, Leiden, The Netherlands, 13–14 September 1991. [Google Scholar]
  53. Geraci, J.R.; Lounsbury, V.J. Marine Mammals Ashore: A Field Guide to Strandings: Texas A&M University Sea Grant Program; Publication TAMU-SG-93-601; Wildlife Disease Association: Lawrence, KS, USA, 1993. [Google Scholar]
  54. Vázquez, J.A.; De la Fuente, J.; Martínez-Cedeira, J.A.; Fernández, C.; Gozalbes, P.; López, A.; Arbelo, M. Documento Técnico Sobre Protocolo Nacional de Actuación para Varamientos de Cetáceos, Informe Realizado para el Ministerio de Agricultura, Alimentación y Medio Ambiente. 2015.
  55. Murch, S.J.; Cox, P.A.; Banack, S.A.; Steele, J.C.; Sacks, O.W. Occurrence of β-methylamino-l-alanine (BMAA) in ALS/PDC patients from Guam. Acta Neurol. Scand. 2004, 110, 267–269. [Google Scholar] [CrossRef] [PubMed]
  56. Masseret, E.; Banack, S.; Boumédiène, F.; Abadie, E.; Brient, L.; Pernet, F.; Juntas-Morales, R.; Pageot, N.; Metcalf, J.; Cox, P.; et al. Dietary BMAA exposure in an Amyotrophic Lateral Sclerosis Cluster from southern France. PLoS ONE 2013, 8, e83406. [Google Scholar] [CrossRef] [PubMed][Green Version]
  57. Lage, S.; Burian, A.; Rasmussen, U.; Costa, P.R.; Annadotter, H.; Godhe, A.; Rydberg, S. BMAA extraction of cyanobacteria samples: Which method to choose? Environ. Sci. Pollut. Res. 2016, 23, 338–350. [Google Scholar] [CrossRef] [PubMed]
  58. Lage, S.; Ström, L.; Godhe, A.; Rydberg, S. Kinetics of β-N-methylamino-L-alanine (BMAA) and 2, 4-diaminobutyric acid (DAB) production by diatoms: The effect of nitrogen. Eur. J. Phycol. 2019, 54, 115–125. [Google Scholar] [CrossRef]
  59. Wenzl, T.; Haedrich, J.; Schaechtele, A.; Robouch, P.; Stroka, J. Guidance Document on the Estimation of LOD and LOQ for Measurements in the Field of Contaminants in Feed and Food; Publications Office of the European Union: Luxembourg, 2016; Volume 58. [Google Scholar] [CrossRef]
  60. Lage, S.; Annadotter, H.; Rasmussen, U.; Rydberg, S. Biotransfer of β-N-Methylamino-l-alanine (BMAA) in a eutrophicated freshwater lake. Mar. Drugs 2015, 13, 1185–1201. [Google Scholar] [CrossRef][Green Version]
  61. Błaszczyk, A.; Siedlecka-Kroplewska, K.; Woźniak, M.; Mazur-Marzec, H. Presence of ß-N-methylamino-L-alanine in cyanobacteria and aquatic organisms from waters of Northern Poland; BMAA toxicity studies. Toxicon 2021, 194, 90–97. [Google Scholar] [CrossRef]
  62. Karlsson, O.; Berg, C.; Brittebo, E.B.; Lindquist, N.G. Retention of the cyanobacterial neurotoxin β-N-methylamino-l-alanine in melanin and neuromelanin-containing cells—A possible link between Parkinson-dementia complex and pigmentary retinopathy. Pigment Cell Melanoma Res. 2009, 22, 120–130. [Google Scholar] [CrossRef]
  63. Masuoka, D.T.; Alcaraz, A.F.; Cohen, M.B.; Spolter, L. Acute distribution of 14C-amino acids in mice as determined by whole-body autoradiography: Adjunct for radio-pharmaceutical synthesis. Int. J. Appl. Radiat. Isot. 1973, 24, 705–706. [Google Scholar] [CrossRef]
  64. Karlsson, O.; Jiang, L.; Andersson, M.; Ilag, L.L.; Brittebo, E.B. Protein association of the neurotoxin and non-protein amino acid BMAA (β-N-methylamino-l-alanine) in the liver and brain following neonatal administration in rats. Toxicol. Lett. 2014, 226, 1–5. [Google Scholar] [CrossRef]
  65. Jiao, Y.; Chen, Q.; Chen, X.; Wang, X.; Liao, X.; Jiang, L.; Wu, J.; Yang, L. Occurrence and transfer of a cyanobacterial neurotoxin β-methylamino-l-alanine within the aquatic food webs of Gonghu Bay (Lake Taihu, China) to evaluate the potential human health risk. Sci. Total Environ. 2014, 468–469, 457–463. [Google Scholar] [CrossRef]
  66. Hammerschlag, N.; Davis, D.A.; Mondo, K.; Seely, M.S.; Murch, S.J.; Glover, W.B.; Divoll, T.; Evers, D.C.; Mash, D.C. Cyanobacterial neurotoxin BMAA and mercury in sharks. Toxins 2016, 8, 238. [Google Scholar] [CrossRef] [PubMed][Green Version]
  67. Mondo, K.; Glover, W.B.; Murch, S.J.; Liu, G.; Cai, Y.; Davis, D.A.; Mash, D.C. Environmental neurotoxins β-N-methylamino-l-alanine (BMAA) and mercury in shark cartilage dietary supplements. Food Chem. Toxicol. 2014, 70, 26–32. [Google Scholar] [CrossRef] [PubMed]
  68. Faassen, E.J.; Antoniou, M.G.; Beekman-Lukassen, W.; Blahova, L.; Chernova, E.; Christophoridis, C.; Combes, A.; Edwards, C.; Fastner, J.; Harmsen, J.; et al. A collaborative evaluation of LC-MS/MS based methods for BMAA analysis: Soluble bound BMAA found to be an important fraction. Mar. Drugs 2016, 14, 45. [Google Scholar] [CrossRef] [PubMed][Green Version]
  69. Murch, S.J.; Cox, P.A.; Banack, S.A. A mechanism for slow release of biomagnified cyanobacterial neurotoxins and neurodegenerative disease in Guam. Proc. Natl. Acad. Sci. USA 2004, 101, 12228–12231. [Google Scholar] [CrossRef] [PubMed][Green Version]
  70. Jeong, Y.; Xue, J.; Park, K.J.; Kannan, K.; Moon, H.-B. Tissue-specific accumulation and body burden of parabens and their metabolites in small cetaceans. Environ. Sci. Technol. 2019, 53, 475–481. [Google Scholar] [CrossRef]
  71. Tilstone, G.H.; Figueiras, F.G.; Lorenzo, L.M. Phytoplankton composition, photosynthesis and primary production during different hydrographic conditions at the Northwest Iberian upwelling system. Mar. Ecol. Prog. Ser. 2003, 252, 89–104. [Google Scholar] [CrossRef][Green Version]
  72. Lorenzo, L.M.; Arbones, B.; Tilstone, G.H.; Figueiras, F.G. Across-shelf variability of phytoplankton composition, photosynthetic parameters and primary production in the NW Iberian upwelling system. J. Mar. Syst. 2005, 54, 157–173. [Google Scholar] [CrossRef]
  73. Villamaña, M.; Marañón, E.; Cermeño, P.; Estrada, M.; Fernández-Castro, B.; Figueiras, F.G.; Latasa, M.; Otero-Ferrer, J.L.; Reguera, B.; Mouriño-Carballido, B. The role of mixing in controlling resource availability and phytoplankton community composition. Prog. Oceanogr. 2019, 178, 102181. [Google Scholar] [CrossRef]
  74. Gallego, J.R.; González-Rojas, E.; Peláez, A.I.; Sánchez, J.; García-Martínez, M.J.; Ortiz, J.E.; Torres, T.; Llamas, J.F. Natural attenuation and bioremediation of Prestige fuel oil along the Atlantic coast of Galicia (Spain). Org. Geochem. 2006, 37, 1869–1884. [Google Scholar] [CrossRef]
  75. Calvo, S.M.; Dosil, J.; del Carmen López Rodríguez, M.; Criado, I.B.; Cremades, J. Checklist of the benthic marine and brackish Galician algae (NW Spain). An. Jardín Botánico Madr. 2005, 62, 69–100. [Google Scholar] [CrossRef]
  76. European Union Commission Regulation(EC) No 853/2004 of the European Parliament and of the Council of 29 April 2004 Laying Down Specific Hygiene Rules for on the Hygiene of Foodstuffs 2004. Available online: (accessed on 23 December 2021).
Figure 1. Location of stranded marine mammals collected along Galician coast that were used in the present study.
Figure 1. Location of stranded marine mammals collected along Galician coast that were used in the present study.
Jmse 10 00314 g001
Figure 2. LC–MS/MS chromatograms of liver tissue precipitated bound sample extract spiked with 5 ng−1 mL of BMAA. The selected reaction monitoring (SRM) transitions 459.1 > 119.1 (common to BMAA and its isomers) and 459.1 > 258.1 (BMAA diagnostic fragment) are shown.
Figure 2. LC–MS/MS chromatograms of liver tissue precipitated bound sample extract spiked with 5 ng−1 mL of BMAA. The selected reaction monitoring (SRM) transitions 459.1 > 119.1 (common to BMAA and its isomers) and 459.1 > 258.1 (BMAA diagnostic fragment) are shown.
Jmse 10 00314 g002
Table 1. Individual data of animals analysed for BMAA, DAB, and AEG. For each animal, samples of liver, kidney, and muscle were collected and analysed. Degradation state: 1 = found alive, dying immediately afterwards, 2 = freshly dead, 3 = moderate decomposition, 4 = advanced decomposition, 5 = skeletal remains. F = female, M = male.
Table 1. Individual data of animals analysed for BMAA, DAB, and AEG. For each animal, samples of liver, kidney, and muscle were collected and analysed. Degradation state: 1 = found alive, dying immediately afterwards, 2 = freshly dead, 3 = moderate decomposition, 4 = advanced decomposition, 5 = skeletal remains. F = female, M = male.
Sample IDSpeciesData of Collection (yyyy/mm/dd)Location (Locality and Coordinates)Size (cm) *Female (F)/Male (M)Degradation StateObservations
DDE194D. delphis2017/07/16Porto do Son (42.68131, −9.03000) 194F3No signs of bycatch
DDE195D. delphis2017/07/16Ribeira (42.56142222, −8.987644444)195M3No signs of bycatch
DDE189D. delphis2017/07/31Cangas (42.24911, −8.79091)189F3Signs of bycatch
DDE182D. delphis2017/07/31Nigrán (42.14245, −8.83796)182M3Three broken ribs and subepidermic hematoma
DDE173D. delphis2018/06/09Ribeira (42.57156, −9.07536)173M1No signs of bycatch
DDE156D. delphis2018/11/07Vigo (42.19025, −8.80904)156M2-
DDE124D. delphis2018/11/25O Grove (42.45576667, −8.921633333)124F3Signs of bycatch
DDE172D. delphis2018/12/17Vigo (42.22298056, −8.772766667)172F3Signs of bycatch
PPH153P. phocoena2009/11/25Fisterra (42.941181, −9.231806)153F3Good aspect
PPH127P. phocoena2011/02/07Fisterra (42.908431, −9.258883)127M3Signs of bycatch
PPH104P. phocoena2011/05/27Baiona (43.056527, −9.295471)104F2Signs of bycatch
PPH137P. phocoena2012/12/02Arteixo (43.316153, −8.534233)137F3Signs gulls and shark bites
PPH142P. phocoena2013/10/15O Grove (43.463064, −8.332825)142F3-
PPH159P. phocoena2014/02/28Ferrol (43.540556, −8.298353)159M3Signs of bycatch
PPH131P. phocoena2015/01/16Cangas (42.261026, −8.849869)131M3Signs of bycatch
PPH160P. phocoena2017/02/20Carballo (43.456950, −8.673272)160M3Signs of bycatch
TTR258T. truncatus2015/10/30Ribeira (42.523304, −9.014026)258M3Stomach with food and Anisakis, ulcers
TTR314T. struncatus2015/12/30Vilanova de Arousa
(42.570726, −8.830053)
314F2Empty stomach
TTR286T. truncatus2016/09/02Cangas (42.295522, −8.820978 286F3Pregnant
TTR150.5T. truncatus2016/09/11Rianxo (42.64833056, −8.8258)150.5M3Signs of aggressions (possible infanticide)
TTR275T. truncatus2017/06/19Foz (43.5665472, −7.2546361)275F2Skinny
TTR138T. truncatus2018/11/30Muros (42.7747527, −9.05476)−138M4Signs of bycatch
* The sign (-) indicates the animal is not entire, and the size is at least the indicated measure.
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Share and Cite

MDPI and ACS Style

Soliño, L.; Kim, S.-Y.; López, A.; Covelo, P.; Rydberg, S.; Costa, P.R.; Lage, S. No β-N-Methylamino-L-alanine (BMAA) Was Detected in Stranded Cetaceans from Galicia (North-West Spain). J. Mar. Sci. Eng. 2022, 10, 314.

AMA Style

Soliño L, Kim S-Y, López A, Covelo P, Rydberg S, Costa PR, Lage S. No β-N-Methylamino-L-alanine (BMAA) Was Detected in Stranded Cetaceans from Galicia (North-West Spain). Journal of Marine Science and Engineering. 2022; 10(3):314.

Chicago/Turabian Style

Soliño, Lucía, Sea-Yong Kim, Alfredo López, Pablo Covelo, Sara Rydberg, Pedro Reis Costa, and Sandra Lage. 2022. "No β-N-Methylamino-L-alanine (BMAA) Was Detected in Stranded Cetaceans from Galicia (North-West Spain)" Journal of Marine Science and Engineering 10, no. 3: 314.

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop