Cyanotoxins Occurrence in Portugal: A New Report on Their Recent Multiplication
Abstract
:1. Introduction
2. Results
2.1. Bloom Frequency and Composition
2.2. Microcystins
2.3. Cylindrospermopsins
2.4. Anatoxin-a
2.5. Saxitoxins
2.6. DNA Sequencing
3. Discussion
4. Conclusions
5. Materials and Methods
5.1. Sampling
5.2. DNA Extraction
5.3. PCR Amplifications
5.4. DNA Sequencing
5.5. ELISA Immunological Assays
Author Contributions
Funding
Conflicts of Interest
References
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Cyanotoxins | MC | CYN | ATX | SXT | |||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Assessment | PCR | ELISA (µg/L) | PCR | ELISA (µg/L) | PCR | ELISA (µg/L) | PCR | ELISA (µg/L) | |||||||||||
Location | mcyA | mcyB | mcyC | mcyD | mcyE | mcyG | cyrA | cyrB | cyrC | cyrJ | anaC | anab/anac | sxtA | sxtG | sxtI | ||||
River Tâmega | |||||||||||||||||||
April | + | - | - | + | - | - | 0.3 | - | - | - | - | ND | - | - | 0.2 | - | - | - | 0.0 |
May | + | - | - | + | + | - | 0.2 | - | - | - | - | ND | - | - | 0.7 | - | - | - | 0.1 |
June | + | - | + | - | + | - | 0.5 | - | - | + | - | ND | - | - | 0.2 | - | - | - | 0.1 |
July | + | + | + | + | + | + | 0.4 | - | - | - | - | ND | + | - | 0.3 | - | + | + | 0.1 |
August | + | + | + | + | + | + | 18.8 | - | - | - | - | ND | + | - | 1.0 | - | + | + | 1.1 |
September | + | + | + | + | + | + | 17.0 | - | - | - | - | ND | + | - | 0.8 | - | + | - | 0.1 |
Torrão Reservoir | |||||||||||||||||||
April | + | + | + | + | - | - | 2.7 | - | - | + | - | ND | + | - | 0.3 | - | - | + | 0.1 |
May | - | - | - | - | - | - | 0.3 | - | - | - | - | ND | + | - | 1.0 | - | - | - | 0.1 |
June | + | - | + | - | + | - | 0.3 | - | - | + | - | ND | - | - | 0.4 | - | - | - | 0.1 |
July | + | + | + | + | + | + | 0.2 | - | - | + | - | ND | + | - | 0.2 | - | + | + | 0.3 |
August | + | + | + | + | + | + | 16.0 | - | - | - | - | ND | + | - | 0.3 | - | + | + | 0.7 |
September | + | + | + | + | + | + | 5.0 | - | + | - | - | ND | + | - | 0.2 | - | + | + | 0.2 |
Porto City Lake 1 | |||||||||||||||||||
April | + | + | + | + | - | - | 4.3 | - | - | - | - | ND | - | - | 5.2 | - | - | − | 0.2 |
May | + | + | + | + | + | - | 11.3 | - | - | - | - | ND | + | - | 2.1 | - | - | - | 0.3 |
June | + | + | + | + | + | - | 3.5 | - | + | - | - | ND | + | - | 6.8 | - | - | - | 0.2 |
July | + | + | + | + | + | + | 1.6 | - | - | - | - | ND | - | - | 0.4 | - | + | - | 0.1 |
August | + | + | + | + | + | + | 2.1 | - | - | - | - | ND | + | - | 1.9 | - | + | + | 0.3 |
September | + | + | + | + | + | + | 2.6 | - | - | - | - | ND | + | - | 1.6 | - | + | + | 0.5 |
Porto City Lake 2 | |||||||||||||||||||
April | - | - | + | - | - | - | 0.3 | - | - | - | - | ND | - | - | 0.2 | - | - | - | 0.2 |
May | + | + | + | + | + | - | 2.0 | - | - | - | - | ND | - | - | 0.4 | - | - | + | 0.2 |
June | + | + | + | + | + | - | 13.3 | - | - | - | - | ND | - | - | 1.5 | - | - | - | 0.3 |
July | + | - | + | + | + | + | 3.9 | - | - | + | - | ND | + | - | 0.4 | - | + | + | 0.9 |
August | + | + | + | + | + | + | 4.8 | - | - | + | - | ND | + | - | 0.9 | - | + | + | 4.3 |
September | + | + | + | + | + | + | 6.5 | - | - | + | - | ND | + | - | 0.6 | - | + | + | 2.6 |
Porto City Lake 3 | |||||||||||||||||||
April | + | - | - | + | - | - | 0.3 | - | - | - | - | ND | - | - | 1.8 | - | - | - | 0.1 |
May | - | - | - | - | - | - | 0.2 | - | - | - | - | ND | - | - | 0.4 | - | - | - | 0.2 |
June | - | - | + | - | - | - | 0.1 | - | - | - | - | ND | - | - | 1.0 | - | - | - | 0.2 |
July | + | - | + | + | + | + | 0.2 | - | - | - | - | ND | - | - | 1.1 | - | + | - | 0.2 |
August | + | + | + | + | + | + | 0.4 | - | - | + | - | ND | - | - | 0.3 | - | + | - | 0.2 |
September | + | + | + | + | + | + | 1.9 | - | - | + | - | ND | - | - | 0.4 | - | + | + | 0.2 |
Mira Lagoon | |||||||||||||||||||
April | - | - | - | - | - | - | 0.5 | - | - | - | - | ND | - | - | 0.4 | - | - | - | 0.1 |
May | - | - | - | - | - | - | 0.3 | - | - | - | - | ND | - | - | 0.5 | - | - | + | 0.2 |
June | - | - | - | - | - | - | 0.3 | - | - | + | - | ND | - | - | 0.5 | - | - | + | 0.6 |
July | + | + | + | + | + | - | 0.6 | - | - | + | - | ND | - | - | 2.1 | - | - | + | 0.3 |
August | + | + | + | + | + | - | 0.3 | - | - | - | - | ND | - | - | 0.6 | - | - | + | 0.5 |
September | + | + | + | + | + | + | 1.9 | - | - | + | - | ND | + | - | 1.0 | - | + | + | 1.3 |
Vela Lagoon | |||||||||||||||||||
April | - | - | - | - | - | - | 0.3 | - | - | - | - | 0.1 | - | - | 1.6 | - | - | - | 1.9 |
May | + | - | - | - | - | - | 0.3 | - | - | - | - | 0.1 | - | - | 0.5 | - | - | + | 1.4 |
June | - | - | - | - | - | - | 0.4 | - | - | + | - | 0.1 | - | - | 1.7 | - | - | + | 1.8 |
July | + | + | + | + | + | + | 2.1 | - | - | + | - | 0.0 | - | - | 0.9 | - | + | - | 1.9 |
August | + | - | - | + | + | + | 0.9 | - | - | - | - | 0.1 | + | - | 0.8 | - | + | + | 1.8 |
September | + | - | - | + | - | + | 8.7 | - | - | - | - | 0.0 | - | - | 1.1 | - | + | - | 2.4 |
Geography | Sampling Site | Trophic Status | Ecosystem Uses | Socio-economic Activities |
---|---|---|---|---|
North Region | River Tâmega | Eutrophic | Recreational | Sports, Fishing |
Torrão Reservoir | Eutrophic | Water provision, irrigation, recreational | Agriculture, Sports, Tourism | |
Porto City Park Lake 1 | Eutrophic | Recreational | Tourism | |
Porto City Park Lake 2 | Eutrophic | Recreational | Tourism | |
Porto City Park Lake 3 | Mesotrophic | Recreational | Tourism | |
Center Region | Mira Lagoon | Eutrophic | Irrigation, Recreational | Agriculture, Sports, Tourism |
Vela Lagoon | Eutrophic | Irrigation, Recreational | Agriculture, Sports, Tourism |
Target | Primer | Primer Sequence 5′-3′ | Reference |
---|---|---|---|
mcyA | mcyA-CD1F | AAAATTAAAAGCCGTATCAAA | [8] |
mcyA-CD1R | AAAAGTGTTTTATTAGCGGCTCAT | ||
mcyB | mcyB 2156-F | ATCACTTCAATCTAACGACT | [14] |
mcyB 3111-R | AGTTGCTGCTGTAAGAAA | ||
mcyC | PSCF1 | GCAACATCCCAAGAGCAAAG | [15] |
PSCR1 | CCGACAACATCACAAAGGC | ||
mcyD | PKDF1 | GACGCTCAAATGATGAAAC | [15] |
PKDR1 | GCAACCGATAAAAACTCCC | ||
mcyE | PKEF1 | CGCAAACCCGATTTACAG | [15] |
PKER1 | CCCCTACCATCTTCATCTTC | ||
mcyG | PKGF1 | ACTCTCAAGTTATCCTCCCTC | [15] |
PKGR1 | AATCGCTAAAACGCCACC | ||
cyrA | AMT Fw | ATTGTAAATAGCTGGAATGAGTGG | [9] |
AMT Rev | TTAGGGAAGTAATCTTCACAG | ||
cyrB | M13 | GGCAAATTGTGATAGCCACGAGC | [7] |
M14 | GATGGAACATCGCTCACTGGTG | ||
cyrC | K18 | CCTCGCACATAGCCATTTGC | [7] |
M4 | GAAGCTCTGGAATCCGGTAA | ||
cyrJ | cynsulfF | ACTTCTCTCCTTTCCCTATC | [11] |
cylnamR | GAGTGAAAATGCGTAGAACTTG | ||
anaC | anaC-genF | TCTGGTATTCAGTCCCCTCTAT | [12] |
anaC-genR | CCCAATAGCCTGTCATCAA | ||
Dolichospermum sp. anaC | anaC-anabF | GCCCGATATTGAAACAAGT | [12] |
anaC-anabR | CACCCTCTGGAGATTGTTTA | ||
sxtA | sxtA855_F | GACTCGGCTTGTTGCTTCCCC | [13] |
sxtA1480_R | GCCAAACTCGCAACAGGAGAAGG | ||
sxtG | sxtG432_F | AATGGCAGATCGCAACCGCTAT | [13] |
sxtG928_R | ACATTCAACCCTGCCCATTCACT | ||
sxtI | sxtI 682F | GGATCTCAAAGAAGATGGCA | [10] |
sxtI 877R | GCCAAACGCAGTACCACTT |
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Moreira, C.; Gomes, C.; Vasconcelos, V.; Antunes, A. Cyanotoxins Occurrence in Portugal: A New Report on Their Recent Multiplication. Toxins 2020, 12, 154. https://doi.org/10.3390/toxins12030154
Moreira C, Gomes C, Vasconcelos V, Antunes A. Cyanotoxins Occurrence in Portugal: A New Report on Their Recent Multiplication. Toxins. 2020; 12(3):154. https://doi.org/10.3390/toxins12030154
Chicago/Turabian StyleMoreira, Cristiana, Cidália Gomes, Vitor Vasconcelos, and Agostinho Antunes. 2020. "Cyanotoxins Occurrence in Portugal: A New Report on Their Recent Multiplication" Toxins 12, no. 3: 154. https://doi.org/10.3390/toxins12030154