Validation of ELISA methods for search and quantification of β-n-methylamino-l-alanine in water and fish tissue

Clausi, Maria Teresa; Vita, Valeria; Bruno, Marco; Franchino, Cinzia; Trifirò, Gianluca; Palumbo, Maria Paola; Floridi, Francesca; Pace, Rita De

doi:10.1080/03067319.2016.1249480

Cited by 10 publications

(5 citation statements)

References 31 publications

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“…False positives were observed in 5 of the 16 MilliQ blank samples measured among the five kits. Given the mixed results of the BMAA ELISA kit in the literature, it cannot be concluded with certainty that BMAA was present in these samples [ 38 , 39 , 40 ].…”

Section: Resultsmentioning

confidence: 99%

“…Notwithstanding the uncertainty involved with and variable performance of the five ELISA kits in numerous studies [ 38 , 39 , 45 , 46 , 47 , 48 , 49 ], they were selected for such practical considerations as availability for the desired analytes and ease of use.…”

Section: Methodsmentioning

confidence: 99%

“…On the other hand, the saxitoxin ELISA kit is functional only from a presence/absence perspective [ 45 ]. Lastly, Faassen et al [ 39 ] reported many likely false positives with the BMAA ELISA kit, but more recently, Clausi et al [ 38 ] reported spiked recoveries in the range of 70–83%, which is adequate. Because this study is the first exploration of these five cyanotoxins across treatment trains in the drinking water plants, the weaknesses in the ELISA measurements were accepted but must be considered in the data interpretation.…”

Section: Methodsmentioning

confidence: 99%

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Cyanotoxins and Cyanobacteria Cell Accumulations in Drinking Water Treatment Plants with a Low Risk of Bloom Formation at the Source

Almuhtaram

Cui

Zamyadi

et al. 2018

Toxins

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Toxic cyanobacteria have been shown to accumulate in drinking water treatment plants that are susceptible to algal blooms. However, the risk for plants that do not experience algal blooms, but that receive a low influx of cells, is not well known. This study determined the extent of cell accumulation and presence of cyanotoxins across the treatment trains of four plants in the Great Lakes region. Samples were collected for microscopic enumeration and enzyme-linked immunosorbent assay (ELISA) measurements for microcystins, anatoxin-a, saxitoxin, cylindrospermopsin, and β-methylamino-L-alanine (BMAA). Low cell influxes (under 1000 cells/mL) resulted in significant cell accumulations (over 1 × 105 cells/mL) in clarifier sludge and filter backwash samples. Microcystins peaked at 7.2 µg/L in one clarifier sludge sample, exceeding the raw water concentration by a factor of 12. Anatoxin-a was detected in the finished drinking water of one plant at 0.6 µg/L. BMAA may have been detected in three finished water samples, though inconsistencies among the BMAA ELISAs call these results into question. In summary, the results show that plants receiving a low influx of cells can be at risk of toxic cyanobacterial accumulation, and therefore, the absence of a bloom at the source does not indicate the absence of risk.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Cyanotoxins and Cyanobacteria Cell Accumulations in Drinking Water Treatment Plants with a Low Risk of Bloom Formation at the Source

Almuhtaram

Cui

Zamyadi

et al. 2018

Toxins

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“…The samples were analysed using ELISA kits from Abraxis (Microcystins ADDA–ELISA Microtiter Plate for MCs and BMAA kit), after a slow freeze–thaw cycle followed by ultrasonic treatment (ELMA S 10, Elmasonic, Singen, Germany), according to the manufacturer’s protocols. Despite initial criticisms of its reliability [ 65 ], the Abraxis BMAA kit was positively validated in recent studies [ 66 ]. The absorbance was read using a Mini photometer (model 6+, Metertech Inc., Taipei, Taiwan).…”

Section: Methodsmentioning

confidence: 99%

Paralytic Shellfish Toxins and Cyanotoxins in the Mediterranean: New Data from Sardinia and Sicily (Italy)

Lugliè

Giacobbe

Riccardi³

et al. 2017

Microorganisms

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Harmful algal blooms represent a severe issue worldwide. They affect ecosystem functions and related services and goods, with consequences on human health and socio-economic activities. This study reports new data on paralytic shellfish toxins (PSTs) from Sardinia and Sicily (Italy), the largest Mediterranean islands where toxic events, mainly caused by Alexandrium species (Dinophyceae), have been ascertained in mussel farms since the 2000s. The toxicity of the A. minutum, A. tamarense and A. pacificum strains, established from the isolation of vegetative cells and resting cysts, was determined by high performance liquid chromatography (HPLC). The analyses indicated the highest toxicity for A. pacificum strains (total PSTs up to 17.811 fmol cell−1). The PSTs were also assessed in a strain of A. tamarense. The results encourage further investigation to increase the knowledge of toxic species still debated in the Mediterranean. This study also reports new data on microcystins (MCs) and β-N-methylamino-l-alanine (BMAA) from a Sardinian artificial lake (Lake Bidighinzu). The presence of MCs and BMAA was assessed in natural samples and in cell cultures by enzyme-linked immunosorbent assay (ELISA). BMAA positives were found in all the analysed samples with a maximum of 17.84 µg L−1. The obtained results added further information on cyanotoxins in Mediterranean reservoirs, particularly BMAA, which have not yet been thoroughly investigated.

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“…The BMAA detection literature is supported by several non-chromatography methods (Clausi et al 2016 ; Faassen et al 2013 ; Kerrin et al 2017 ; Beri et al 2018 ). However, we note that the currently available commercial ELISA is considered a flawed test by the scientific community generating many false positives and is not a good research tool in its present form (Faassen et al 2013 ; Bláhová et al 2017 ).…”

Section: The Detection and Quantification Of Bmaamentioning

confidence: 99%

Is Exposure to BMAA a Risk Factor for Neurodegenerative Diseases? A Response to a Critical Review of the BMAA Hypothesis

Dunlop

et al. 2021

Neurotox Res

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In a literature survey, Chernoff et al. (2017) dismissed the hypothesis that chronic exposure to β-N-methylamino-L-alanine (BMAA) may be a risk factor for progressive neurodegenerative disease. They question the growing scientific literature that suggests the following: (1) BMAA exposure causes ALS/PDC among the indigenous Chamorro people of Guam; (2) Guamanian ALS/PDC shares clinical and neuropathological features with Alzheimer’s disease, Parkinson’s disease, and ALS; (3) one possible mechanism for protein misfolds is misincorporation of BMAA into proteins as a substitute for L-serine; and (4) chronic exposure to BMAA through diet or environmental exposures to cyanobacterial blooms can cause neurodegenerative disease. We here identify multiple errors in their critique including the following: (1) their review selectively cites the published literature; (2) the authors reported favorably on HILIC methods of BMAA detection while the literature shows significant matrix effects and peak coelution in HILIC that may prevent detection and quantification of BMAA in cyanobacteria; (3) the authors build alternative arguments to the BMAA hypothesis, rather than explain the published literature which, to date, has been unable to refute the BMAA hypothesis; and (4) the authors erroneously attribute methods to incorrect studies, indicative of a failure to carefully consider all relevant publications. The lack of attention to BMAA research begins with the review’s title which incorrectly refers to BMAA as a “non-essential” amino acid. Research regarding chronic exposure to BMAA as a cause of human neurodegenerative diseases is emerging and requires additional resources, validation, and research. Here, we propose strategies for improvement in the execution and reporting of analytical methods and the need for additional and well-executed inter-lab comparisons for BMAA quantitation. We emphasize the need for optimization and validation of analytical methods to ensure that they are fit-for-purpose. Although there remain gaps in the literature, an increasingly large body of data from multiple independent labs using orthogonal methods provides increasing evidence that chronic exposure to BMAA may be a risk factor for neurological illness.

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Validation of ELISA methods for search and quantification of β-n-methylamino-l-alanine in water and fish tissue

Cited by 10 publications

References 31 publications

Cyanotoxins and Cyanobacteria Cell Accumulations in Drinking Water Treatment Plants with a Low Risk of Bloom Formation at the Source

Cyanotoxins and Cyanobacteria Cell Accumulations in Drinking Water Treatment Plants with a Low Risk of Bloom Formation at the Source

Paralytic Shellfish Toxins and Cyanotoxins in the Mediterranean: New Data from Sardinia and Sicily (Italy)

Is Exposure to BMAA a Risk Factor for Neurodegenerative Diseases? A Response to a Critical Review of the BMAA Hypothesis

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