cyanotoxin producers in Greek lakes: towards identifying them
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1 Spyros Gkelis School of Biology, Aristotle University of Thessaloniki [AUTH] Sevilla, February 19th 2015 cyanotoxin producers in Greek lakes: towards identifying them
2 cyanotoxin producers in Greek lakes: towards identifying them i. peptide diversity and concentration ii. microcystin producers and the environment iii. cyanotoxin producers: field studies iv. cyanotoxin producers: strains v. other metabolites, other environments
3 peptide diversity and concentration microcystins (MCs) 100% samples MC-LR [D-Asp 3 ]MC-LR MC-RR [Dha 7 ]MC-RR MC-YR MC-LA blooms Gkelis et al. 2005, Environ. Toxicol.
4 peptide diversity and concentration unedintified MC U1, U2, U3, U4 mau mau mau U1 238nm U3 238nm min Lake Kerkini meeting Cyanocost U2 238nm 2015, Sevilla U4 238nm Gkelis, Gkelis et al. 2005, Environ. Toxicol.Spyros nm nm blooms
5 peptide diversity and concentration MC TMC μg g -1 dw. TMC ( ) και TAC ( ) (μg g -1 ) blooms Β Κε Μ Ζ Κα Π Α Gkelis et al. 2005, Environ. Toxicol.
6 peptide diversity and concentration MC-LR MC-RR MC-YR U1 U2 σχετική συγκέντρωση μικροκυστίνης (%) relevant MC concentration (%) MC-LR MC-RR [D-Asp 3 ] MC-LR [Dha 7 ] MC-RR MC-YR n: (26) (25) (3) (4) (11) (2) (10) (8) (4) (7) MC-LA U1 U2 U3 blooms U4 Gkelis et al. 2005, Environ. Toxicol.
7 peptide diversity and concentration 40 ο Ν 30 ο Ν ο Ν MC-LR MC-RR MC-YR Japan Park et al. 1993a Park et al. 1993a S. Κorea Park 2001 Portugal Vasconcelos et al Greece Gkelis et al geographical patterns in MC variants blooms Gkelis et al. 2005, Environ. Toxicol.
8 peptide diversity and concentration microcystins (95%) water
9 peptide diversity and concentration microcystins MC-LR > MC-RR > U1> MC-YR relevant MC concentration (%) σχετική συγκέντρωση (%) n: water MC-LR MC-RR MC-YR U1 U5 (25) (27) (8) (17) (2)
10 peptide diversity and concentration cluster analysis relevant peptide concentration 4 groups of samples water
11 peptide diversity and concentration Group 1 Group 2 Group 3 Group 4 Axis Ι MC-RR MC-RR MC-RR MC-RR predominant MCs Axis IΙ MC-LR MC-LR MC-LR MC-LR Άξονας II (35.1 %) Ομάδα 1 Ομάδα 2 Ομάδα 3 Ομάδα Άξονας I (64.8 %) YR UA LR MC-YR MC-LR water MC-RR strains/species/functional groups RR
12 peptide diversity and concentration Group 1 Group 2 Group 3 Group 4 Axis Ι MC-RR MC-RR MC-RR MC-RR predominant MCs Axis IΙ MC-LR MC-LR MC-LR MC-LR Lake Kastoria Kastoria Κerkini Zazari Pamvotis Volvi Doirani Amvrakia Taxa Microcystis aeruginosa Microcystis aeruginosa Microcystis spp. Microcystis aeruginosa Anabaena flos-aquae Jaaginema subtilissimum Aphanizomenon flos-aquae water strains/species/functional groups
13 total MC concentration (μg L -1 ) συνολική συγκέντρωση μg L -1 peptide diversity and concentration samples > μg L -1 ΣΜC μg L -1 water
14 microcystin producers and environment highly eutrophic shallow, urban Mediterranean Lake Pamvotis NW Greece monthly sampling (14 months, )
15 microcystin producers and environment Gkelis et al. 2014, Harmful Algae physico-chemical parameters temperature ph dissolved oxygen Soluble Reactive Phosphorus Nitrogen (NO 2, N0 3, NH 4 )
16 microcystin producers and environment ELISA cyanotoxin quantification Gkelis et al. 2014, Harmful Algae microcystins (MCs) cylindrospermopsins (CYNs) saxitoxins (STXs) only in two samples
17 microcystin producers and environment X cyanotoxin genes PCR detection cyanobacteria bloom species identification X Gkelis et al. 2014, Harmful Algae mcye sequences
18 microcystin producers and environment Principal Component Analysis Gkelis et al. 2014, Harmful Algae microcystins (MCs) temperature Dissolved Inorganic Nitrogen Soluble Reactive Phosphorus
19 growth rate μ (d -1 ) microcystin producers and environment Gkelis et al. 2014, Harmful Algae could temperature rise promote microcystin production? MC-producing Microcystis AUTH 1410 strain
20 microcystin producers and environment Gkelis et al. 2014, Harmful Algae Long-term limnological data
21 cyanotoxin producers: field study Lakes microcystins (MCs) cylindrospermopsins (CYNs) saxitoxins (STXs) species occurrence/biomass cyanotoxin genes
22 cyanotoxin producers: field study Gkelis and Zaoutsos, 2014, Toxicon 78, 1-9 dominant species Gkelis & Zaoutos 2014, Toxicon genes mcya, mcyb, mcye, mcye/ndaf pks ps stxi
23 cyanotoxin producers: field study Gkelis and Zaoutsos, 2014, Toxicon 78, 1-9 Gkelis & Zaoutos 2014, Toxicon MCs 100% CYNs 39% STXs 15%
24 cyanotoxin producers: field study Anabaenopsis elenkinii Cylindrospermopsis raciborskii Microcystis wesenbergii Microcystis spp. Gkelis & Zaoutos 2014, Toxicon MC < 30 μg L -1 ; STX and CYN presen ps, rpoc1, and sxti genes present 30 μg L -1 < MC < 70 μg L -1 ; no STX or CYL; mcya and sxti genes present MC > 70 μg L -1 ; CYN present; all mcy genes present
25 cyanotoxin producers: strains Chroococcus Synechococcus Jaaginema Anabaena Microcystis Limnothrix Pseudanabaena Calothrix Gkelis et al. 2015, Open Life Sci
26 cyanotoxin producers: strains Gkelis et al. 2015, Open Life Sci
27 cyanotoxin producers: strains Lake Karla, Central Greece Strains isolated Limnothrix redekei Planktothrix agardhii Pseudanabaena Synechococcus Microcystis Radiocystis Anabaena/ Dolichospermum Anabaenopsis elenkinii Sphaerospermopsis aphanizomenoides Cylindrospermopsis raciborskii Lake Karla
28 cyanotoxin producers: field study II Lake Karla, Central Greece dried in the 1960s restored in heavy water blooms : monitoring
29 not only toxins: other metabolites? effects? water and/or methanol extracts antimicrobial activity cancer human/mammal cell lines (HEPG2, Caco2) higher plants (Triticum, Pisum) Escherichia coli Staphylococcus aureus 9518 Pseudomonas aeruginosa Bacillus subtilis
30 not only lakes: sponge-cyanobacteria symbiosis Chondrilla nucula assess the symbiotic cyanobacteria diversity (16S clone libraries) isolate symbiotic cyanobacteria search for new metabolites Picture: D. Kostantinou Graduate student
STRR / LTRR FINGERPRINTING PATTERNS
01 0 20 40 60 80 100 STRR LTRR Oscillatoriales Phormidium sp. Planktothrix rubescens Planktothrix agardhii STRR / LTRR FINGERPRIING PATTERNS 0 r = 0.97 20 40 60 80 100 Phormidium sp. Limnothrix redekei
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