Supplementary Data File S1 Diversity of cyanobacteria and cyanotoxins in Hartbeespoort Dam, South Africa
Andreas Ballot
a,b,*, Morten Sandvik
b, Thomas Rundberget
b, Christo J. Botha
c, Christopher O.
Miles
b*Corresponding author. Tel: +47 98215480; fax: +47 22185200; E-mail address:
andreas.ballot@niva.no
a
Norwegian Institute for Water Research, Gaustadalléen 21, N-0349 Oslo, Norway
b
Norwegian Veterinary Institute, PO Box 750 Sentrum, N-0106 Oslo, Norway
c
Department of Paraclinical Sciences, Faculty of Veterinary Science, University of Pretoria, Private Bag X04, Onderstepoort, 0110, South Africa
Appendix A. Supplementary Data File (74 pp.)
Page S2 LC-MS Chromatograms of extracts of Hartebeespoort Dam and M. aeruginosa AB2011/53
Page S4 MS
2spectra of microcystins. NB weaker peaks display some spurious peaks due to reduced signal-to-noise, and for some very weak peaks, suitable MS
2spectra were not obtained.
Supplementary Material: Marine and Freshwater Research 65, 175-189.
SUPPLEMENTARY MATERIAL
Figure S1a. Chromatograms of Hartbeespoort Dam extract
LC-MS of Hartbeespoort Dam
(A) Underivatised extract
(B) Mercaptoethanol-derivatised extract
(C) Subtracted chromatogram (A minus B)
20 40 60 80 100 20 40 60 80 100
Relative Abundance
20 40 60 80 100
3.92 996.0 3.83 987.9 3.75 1020.5 2.08
1038.8 4.15
1016.5 2.75
945.8 4.95
1048.5 5.37 1044.5 2.80
973.8 4.32
1068.9 7.50
926.4 5.48
1120.4 4.70
966.3
8.18 988.7 5.88
988.6 8.22
988.8 7.84
951.3 2.25
1038.9
8.69 920.1 7.09
904.0 6.26
1045.5 6.67 901.2 3.85
988.1
3.76
1020.6 4.17 1016.4 2.09
1116.9
2.77 945.9
3.50
1127.9 4.95
1048.5 5.37 1044.5 2.82
973.8 4.29
1146.9
7.50 926.6 5.50
1120.4 4.70
990.5
7.79 1066.6 5.90
988.5
8.05 1080.8 2.49
931.7
8.71 902.1 6.29
1023.3
7.19 981.4 6.75
1021.2 3.92
996.0
3.81 1045.8 2.08
1038.8 3.53
1049.8
4.18 1017.4 2.74
945.8 4.32
1068.9 2.78
973.9 8.18
988.7 5.35
1045.6 4.93
1048.5
8.22 988.8 7.47
1045.6 5.45
1043.9
7.89 1104.4 2.58
981.1
8.99 945.3 6.59
1047.7 5.92
893.8
7.16 962.3
LC-MS of Hartbeespoort Dam
(A) Underivatised extract
(B) Mercaptoethanol-derivatised extract
(C) Subtracted chromatogram (A minus B)
20 40 60 80 100 20 40 60 80 100
Relative Abundance
20 40 60 80 100
3.92 996.0 3.83 987.9 3.75 1020.5 2.08
1038.8 4.15
1016.5 2.75
945.8 4.95
1048.5 5.37 1044.5 2.80
973.8 4.32
1068.9 7.50
926.4 5.48
1120.4 4.70
966.3
8.18 988.7 5.88
988.6 8.22
988.8 7.84
951.3 2.25
1038.9
8.69 920.1 7.09
904.0 6.26
1045.5 6.67 901.2 3.85
988.1
3.76
1020.6 4.17 1016.4 2.09
1116.9
2.77 945.9
3.50
1127.9 4.95
1048.5 5.37 1044.5 2.82
973.8 4.29
1146.9
7.50 926.6 5.50
1120.4 4.70
990.5
7.79 1066.6 5.90
988.5
8.05 1080.8 2.49
931.7
8.71 902.1 6.29
1023.3
7.19 981.4 6.75
1021.2 3.92
996.0
3.81 1045.8 2.08
1038.8 3.53
1049.8
4.18 1017.4 2.74
945.8 4.32
1068.9 2.78
973.9 8.18
988.7 5.35
1045.6 4.93
1048.5
8.22 988.8 7.47
1045.6 5.45
1043.9
7.89 1104.4 2.58
981.1
8.99 945.3 6.59
1047.7 5.92
893.8
7.16 962.3
(A) Underivatised extract
(B) Mercaptoethanol-derivatised extract
(C) Subtracted chromatogram (A minus B)
20 40 60 80 100 20 40 60 80 100
Relative Abundance
20 40 60 80 100
3.92 996.0 3.83 987.9 3.75 1020.5 2.08
1038.8 4.15
1016.5 2.75
945.8 4.95
1048.5 5.37 1044.5 2.80
973.8 4.32
1068.9 7.50
926.4 5.48
1120.4 4.70
966.3
8.18 988.7 5.88
988.6 8.22
988.8 7.84
951.3 2.25
1038.9
8.69 920.1 7.09
904.0 6.26
1045.5 6.67 901.2 3.85
988.1
3.76
1020.6 4.17 1016.4 2.09
1116.9
2.77 945.9
3.50
1127.9 4.95
1048.5 5.37 1044.5 2.82
973.8 4.29
1146.9
7.50 926.6 5.50
1120.4 4.70
990.5
7.79 1066.6 5.90
988.5
8.05 1080.8 2.49
931.7
8.71 902.1 6.29
1023.3
7.19 981.4 6.75
1021.2 3.92
996.0
3.81 1045.8 2.08
1038.8 3.53
1049.8
4.18 1017.4 2.74
945.8 4.32
1068.9 2.78
973.9 8.18
988.7 5.35
1045.6 4.93
1048.5
8.22 988.8 7.47
1045.6 5.45
1043.9
7.89 1104.4 2.58
981.1
8.99 945.3 6.59
1047.7 5.92
893.8
7.16 962.3
Figure S1b. Chromatograms of Hartbeespoort Dam extract
LC-MS of AB2011/53 (1.8-5.5 min)
(A) Underivatised extract
(B) Mercaptoethanol-derivatised extract
(C) Subtracted chromatogram (A minus B)
20 40 60 80 100 20 40 60 80 100
Relative Abundance
20 40 60 80 100
3.89 995.9
1.97 906.0
3.77 1045.8 2.03
1038.8 4.11
1016.5 3.72
1020.5
5.34 1044.5 3.52
1049.8 4.92
1048.5 4.29
1068.8 3.30
988.5
5.03 1037.9 2.27
1052.1
2.98 992.5 2.59
999.6
4.56 1030.4 2.66
950.8
4.84 920.6 1.91
906.0 3.75
1073.8
3.70
1123.7 4.06
1016.4 5.30
1044.5 4.88
1048.5 3.40
1127.9 2.84
1225.7
4.21 1146.8 3.26
988.2
4.82 1115.8 2.92
1225.6 2.27
1132.0
4.54 1030.5 2.55
1180.7
5.00 1048.5 3.89
995.9
3.77 1045.8 2.03
1038.8
3.52
1049.8 5.32
1044.5 4.29
1068.8 4.18
1029.8
4.90 1048.5
5.03 1037.9 2.21
1003.6
4.40 1000.8 3.32
987.6
4.78 956.2 2.59
1090.5
2.83 969.7
3.23 1096.8
LC-MS of AB2011/53 (1.8-5.5 min)
(A) Underivatised extract
(B) Mercaptoethanol-derivatised extract
(C) Subtracted chromatogram (A minus B)
20 40 60 80 100 20 40 60 80 100
Relative Abundance
20 40 60 80 100
3.89 995.9
1.97 906.0
3.77 1045.8 2.03
1038.8 4.11
1016.5 3.72
1020.5
5.34 1044.5 3.52
1049.8 4.92
1048.5 4.29
1068.8 3.30
988.5
5.03 1037.9 2.27
1052.1
2.98 992.5 2.59
999.6
4.56 1030.4 2.66
950.8
4.84 920.6 1.91
906.0 3.75
1073.8
3.70
1123.7 4.06
1016.4 5.30
1044.5 4.88
1048.5 3.40
1127.9 2.84
1225.7
4.21 1146.8 3.26
988.2
4.82 1115.8 2.92
1225.6 2.27
1132.0
4.54 1030.5 2.55
1180.7
5.00 1048.5 3.89
995.9
3.77 1045.8 2.03
1038.8
3.52
1049.8 5.32
1044.5 4.29
1068.8 4.18
1029.8
4.90 1048.5
5.03 1037.9 2.21
1003.6
4.40 1000.8 3.32
987.6
4.78 956.2 2.59
1090.5
2.83 969.7
3.23 1096.8
Figure S2a. MS
2spectrum of tentative [DMAdda
5]MC-LA
Hartbeespoort Dam m/z 896, R t 6.18 min
[DMAdda 5 ]MC-LA (tentative)
250 300 350 400 450 500 550 600 650 700 750 800 850 900
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
878.5
877.6 879.5
860.4 776.5
365.4
851.6 833.4 663.3
758.6
366.4 553.4 825.5
799.4 662.4
585.3 599.8 664.4 748.4
523.5 693.5
381.4 402.3 446.4 490.4 880.3
343.4 321.4 278.4
Hartbeespoort Dam m/z 896, R t 6.18 min
[DMAdda 5 ]MC-LA (tentative)
250 300 350 400 450 500 550 600 650 700 750 800 850 900
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
878.5
877.6 879.5
860.4 776.5
365.4
851.6 833.4 663.3
758.6
366.4 553.4 825.5
799.4 662.4
585.3 599.8 664.4 748.4
523.5 693.5
381.4 402.3 446.4 490.4 880.3
343.4 321.4 278.4
Figure S2b. MS
2spectr um of tentative [Asp
3]MC-LA
Hartbeespoort Dam m/z 896, R t 7.50 min [Asp 3 ]MC-LA (tentative)
250 300 350 400 450 500 550 600 650 700 750 800 850 900
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
762.4
745.4
744.6
559.3
375.3 693.4
509.3 860.5
580.3
454.3 661.4 829.5
446.4 798.3
533.5 603.6 673.5 711.5
388.3 481.4 503.4 642.5 876.9
347.4 391.5 439.5 781.5
334.3 305.4
282.3 896.7
Hartbeespoort Dam m/z 896, R t 7.50 min [Asp 3 ]MC-LA (tentative)
250 300 350 400 450 500 550 600 650 700 750 800 850 900
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
762.4
745.4
744.6
559.3
375.3 693.4
509.3 860.5
580.3
454.3 661.4 829.5
446.4 798.3
533.5 603.6 673.5 711.5
388.3 481.4 503.4 642.5 876.9
347.4 391.5 439.5 781.5
334.3 305.4
282.3 896.7
Figure S2c. MS
2spectrum of tentative MC-LA
Hartbeespoort Dam m/z 910, R t 8.29 min MC-LA
250 300 350 400 450 500 550 600 650 700 750 800 850 900
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
776.4
892.5
758.5
878.4 893.5 693.4
559.3 375.3 402.3 468.4 509.4
580.3
446.3 675.5
860.4
485.4 597.4 725.4 809.4
661.3 548.3
331.2 614.4 777.5
314.3 347.3 443.4 712.5 821.6
282.4 909.9
Hartbeespoort Dam m/z 910, R t 8.29 min MC-LA
250 300 350 400 450 500 550 600 650 700 750 800 850 900
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
776.4
892.5
758.5
878.4 893.5 693.4
559.3 375.3 402.3 468.4 509.4
580.3
446.3 675.5
860.4
485.4 597.4 725.4 809.4
661.3 548.3
331.2 614.4 777.5
314.3 347.3 443.4 712.5 821.6
282.4 909.9
Figure S2d. MS
2spectrum of tentative MC-LA ba
Hartbeespoort Dam m/z 924, R t 9.08 min MC-LAba (tentative)
250 300 350 400 450 500 550 600 650 700 750 800 850 900
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
790.4
906.4
772.4
888.5 794.5
446.2 482.3 559.1 693.3
509.3
375.3 416.3 580.3 739.3 795.5 862.6
593.3 755.5 823.3
698.9 689.4
398.5 477.2 628.5
345.2 543.3
426.3 314.4
282.3 925.3
Hartbeespoort Dam m/z 924, R t 9.08 min MC-LAba (tentative)
250 300 350 400 450 500 550 600 650 700 750 800 850 900
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
790.4
906.4
772.4
888.5 794.5
446.2 482.3 559.1 693.3
509.3
375.3 416.3 580.3 739.3 795.5 862.6
593.3 755.5 823.3
698.9 689.4
398.5 477.2 628.5
345.2 543.3
426.3 314.4
282.3 925.3
Figure S2e. MS
2spectrum of tentative MC-YA from Hartbeespoort Dam
Hartbeespoort Dam m/z 960, R t 8.13 min MC-YA (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
826.3
942.4
808.4
943.4 623.3
928.4 452.3
518.3
743.3
446.2 910.5
375.3 393.4 509.3 535.3 580.2 624.4 725.3 874.4
364.2 434.3 477.3 548.2 650.3 711.4 775.4 791.3 827.3
347.3
297.3 944.2
Hartbeespoort Dam m/z 960, R t 8.13 min MC-YA (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
826.3
942.4
808.4
943.4 623.3
928.4 452.3
518.3
743.3
446.2 910.5
375.3 393.4 509.3 535.3 580.2 624.4 725.3 874.4
364.2 434.3 477.3 548.2 650.3 711.4 775.4 791.3 827.3
347.3
297.3 944.2
Figure S2f. MS
2spectrum of [Asp
3, DMAdda
5]MC-L R standard from Nostoc 152
Hydrolysed Nostoc culture 152 m/z 967, R t 2.57 min
[Asp 3 , DMAdda 5 ]MC-LR from hydrolysis of confirmed [Asp 3 , ADMAdda 5 ]MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
939.6
950.5
585.5
830.5 847.5 539.4
557.5
895.7
951.6 538.5
896.6 586.5
456.4
852.6 711.5
375.3 700.5
455.4 521.4 654.5 739.6 755.6 802.6
446.4
383.3 457.4 511.4
368.4 611.5
325.3 650.6 787.5
289.2
967.0
Hydrolysed Nostoc culture 152 m/z 967, R t 2.57 min
[Asp 3 , DMAdda 5 ]MC-LR from hydrolysis of confirmed [Asp 3 , ADMAdda 5 ]MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
939.6
950.5
585.5
830.5 847.5 539.4
557.5
895.7
951.6 538.5
896.6 586.5
456.4
852.6 711.5
375.3 700.5
455.4 521.4 654.5 739.6 755.6 802.6
446.4
383.3 457.4 511.4
368.4 611.5
325.3 650.6 787.5
289.2
967.0
Figure S2g. MS
2spectrum of confirmed [Asp
3, DMAdda
5]MC-LR stand ard from Hartbeespoort Dam
Hartbeespoort Dam m/z 967, R t 2.57 min [Asp 3 , DMAdda 5 ]MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
949.6
948.6
583.3
950.6
922.6 585.5
921.2 904.5 605.4
830.6 902.5
539.5 557.5 835.6 951.5
801.5
538.5 637.4 654.6 669.4 755.6 784.5
385.4 407.4 462.4 496.4 708.5 966.9
353.5 342.5 301.3
Hartbeespoort Dam m/z 967, R t 2.57 min [Asp 3 , DMAdda 5 ]MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
949.6
948.6
583.3
950.6
922.6 585.5
921.2 904.5 605.4
830.6 902.5
539.5 557.5 835.6 951.5
801.5
538.5 637.4 654.6 669.4 755.6 784.5
385.4 407.4 462.4 496.4 708.5 966.9
353.5 342.5 301.3
Figure S2h. MS
2spectrum of tentative [Asp
3, Dha
7]MC-LR standard from Hartbeespoort Dam
Hartbeespoort Dam m/z 967, R t 3.93 min
[Asp 3 , Dha 7 ]MC-LR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
599.4
939.5
950.4
525.4
571.4 524.3
542.4 838.5
600.4
895.5 837.5
951.5 816.5
668.4
918.4 852.4
696.4 508.4
491.4
469.5 714.4
361.3
432.2 625.4 869.5
414.4
333.3 453.4 653.4 735.5 753.4 810.4
289.3
958.0
Hartbeespoort Dam m/z 967, R t 3.93 min
[Asp 3 , Dha 7 ]MC-LR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
599.4
939.5
950.4
525.4
571.4 524.3
542.4 838.5
600.4
895.5 837.5
951.5 816.5
668.4
918.4 852.4
696.4 508.4
491.4
469.5 714.4
361.3
432.2 625.4 869.5
414.4
333.3 453.4 653.4 735.5 753.4 810.4
289.3
958.0
Figure S2i. MS
2spectrum of [DMAdda
5]MC-LR standard from Nostoc 152
Hydrolysed Nostoc culture 152 m/z 981, R t 2.59 min
[DMAdda 5 ]MC-LR from hydrolysis of confirmed [ADMAdda 5 ]MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
953.6
964.5
585.4
553.4
844.5 852.6 557.5
909.6 552.4
965.6 896.5
586.5 937.6
668.5 696.5 714.5
862.6
470.4 824.6
375.3 446.2 455.5 525.5 535.4 611.4 739.5 769.5 816.5
285.3 303.3 347.3 397.3 487.3 636.4
966.1
Hydrolysed Nostoc culture 152 m/z 981, R t 2.59 min
[DMAdda 5 ]MC-LR from hydrolysis of confirmed [ADMAdda 5 ]MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
953.6
964.5
585.4
553.4
844.5 852.6 557.5
909.6 552.4
965.6 896.5
586.5 937.6
668.5 696.5 714.5
862.6
470.4 824.6
375.3 446.2 455.5 525.5 535.4 611.4 739.5 769.5 816.5
285.3 303.3 347.3 397.3 487.3 636.4
966.1
Figure S2j. MS
2spectrum of confirmed [DMAdda
5]MC-LR standard from Hartbeespoort Dam
Hartbeespoort Dam m/z 981, R t 2.59 min [DMAdda 5 ]MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
953.5
963.4 964.5 585.3
553.4
844.5
557.5 861.5
965.5 909.5
918.5
552.4 896.5 936.5
714.4
586.4 862.5
668.4 534.6
696.4
375.3 397.4 455.4 510.4 588.3611.4 654.3 715.4 769.5 797.4816.2
303.3 347.3 446.3 474.6
971.9
Hartbeespoort Dam m/z 981, R t 2.59 min [DMAdda 5 ]MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
953.5
963.4 964.5 585.3
553.4
844.5
557.5 861.5
965.5 909.5
918.5
552.4 896.5 936.5
714.4
586.4 862.5
668.4 534.6
696.4
375.3 397.4 455.4 510.4 588.3611.4 654.3 715.4 769.5 797.4816.2
303.3 347.3 446.3 474.6
971.9
Figure S2k. MS
2spectrum of confirmed [DMAdda
5]MC-LR standard from M. aeruginosa AB2011/53
AB2011/53
m/z 981, R t 2.59 min [DMAdda 5 ]MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
953.6
963.6
585.5 964.6
583.3
553.5 844.6 861.6
909.6 552.6
557.5
965.7
586.5 937.6
714.5 896.6
696.6
869.7
470.5 605.4 668.5 824.6
375.3 399.4 455.5 510.5 535.4 611.5637.4 715.6 769.6 816.6
339.4 809.6
285.3 654.4
982.3
AB2011/53
m/z 981, R t 2.59 min [DMAdda 5 ]MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
953.6
963.6
585.5 964.6
583.3
553.5 844.6 861.6
909.6 552.6
557.5
965.7
586.5 937.6
714.5 896.6
696.6
869.7
470.5 605.4 668.5 824.6
375.3 399.4 455.5 510.5 535.4 611.5637.4 715.6 769.6 816.6
339.4 809.6
285.3 654.4
982.3
Figure S2l. MS
2spectrum of tentative [Asp
3]MC-LR from Hartbeespoort Dam
Hartbeespoort Dam m/z 981, R t 3.87 min [Asp 3 ]MC-LR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
953.5 964.5
599.4
539.4
909.5
965.5 538.4
571.4
910.5 830.4
847.4
600.4 932.4
866.5 714.4
696.4 375.3
668.4 456.4
724.4
455.3 496.4 521.4 625.4 769.5
368.3 397.4 439.3 812.5
289.3 329.3
980.2
Hartbeespoort Dam m/z 981, R t 3.87 min [Asp 3 ]MC-LR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
953.5 964.5
599.4
539.4
909.5
965.5 538.4
571.4
910.5 830.4
847.4
600.4 932.4
866.5 714.4
696.4 375.3
668.4 456.4
724.4
455.3 496.4 521.4 625.4 769.5
368.3 397.4 439.3 812.5
289.3 329.3
980.2
Figure S2m. MS
2spectrum of tentative [Asp
3]MC-LR from M. aeruginosa AB2011/53
AB2011/53
m/z 981, R t 3.87 min [Asp 3 ]MC-LR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
953.6
964.6
599.5
539.5
909.6
965.6 571.5
538.5
910.7 851.6
830.6
932.6 600.5
852.7 883.6 696.6
714.5 456.5
375.4 668.5
469.6
446.3 511.4 769.6
383.3 470.5 623.4 715.5
368.3 439.4 641.5 796.6
347.3 289.3
966.7
AB2011/53
m/z 981, R t 3.87 min [Asp 3 ]MC-LR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
953.6
964.6
599.5
539.5
909.6
965.6 571.5
538.5
910.7 851.6
830.6
932.6 600.5
852.7 883.6 696.6
714.5 456.5
375.4 668.5
469.6
446.3 511.4 769.6
383.3 470.5 623.4 715.5
368.3 439.4 641.5 796.6
347.3 289.3
966.7
Figure S2n. MS
2spectrum of confirmed [Dha
7]MC-LR from Hartbeespoort Dam
Hartbeespoort Dam m/z 981, R t 3.99 min [Dha 7 ]MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
599.4
963.5 953.5
964.5
539.4
538.4
852.5 571.4
600.4 851.5
556.4
965.5
710.4 909.5
830.4 728.4
682.4
522.4 853.5
505.4 824.6
470.4 668.4
869.5 625.4
361.3 432.2 729.5
285.3 333.3 414.3 751.5 797.5
966.4
Hartbeespoort Dam m/z 981, R t 3.99 min [Dha 7 ]MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
599.4
963.5 953.5
964.5
539.4
538.4
852.5 571.4
600.4 851.5
556.4
965.5
710.4 909.5
830.4 728.4
682.4
522.4 853.5
505.4 824.6
470.4 668.4
869.5 625.4
361.3 432.2 729.5
285.3 333.3 414.3 751.5 797.5
966.4
Figure S2o. MS
2spectrum of confirmed [Dha
7]MC-LR from M. aeruginosa AB2011/53
AB2011/53
m/z 981, R t 3.99 min [Dha 7 ]MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
599.5
963.6
953.6 964.6
539.4
852.6 538.4
556.5 600.5 851.6
965.6 909.6
710.5
830.6
728.5 932.6
682.5 853.6
522.5
505.4 824.6
469.5
668.5 869.7
625.5 487.4
432.3
361.3 729.6
285.3 333.3 397.4 641.6 797.5
966.3
AB2011/53
m/z 981, R t 3.99 min [Dha 7 ]MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
599.5
963.6
953.6 964.6
539.4
852.6 538.4
556.5 600.5 851.6
965.6 909.6
710.5
830.6
728.5 932.6
682.5 853.6
522.5
505.4 824.6
469.5
668.5 869.7
625.5 487.4
432.3
361.3 729.6
285.3 333.3 397.4 641.6 797.5
966.3
Figure S2p. MS
2spectrum of confirmed [Asp
3]MC-LY from Hartbeespoort Dam
Hartbeespoort Dam m/z 988, R t 8.15 min [Asp 3 ]MC-LY (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
854.3
837.2
970.4
836.4
559.2
971.4 693.3
956.3 546.3
375.3
446.2 480.3 509.3 580.2
939.3 809.3
463.3 661.2
753.4 545.4
435.3
409.3 713.2 791.4 887.4 911.3
347.3 629.3 657.3
329.3
296.2 776.4 972.2
Hartbeespoort Dam m/z 988, R t 8.15 min [Asp 3 ]MC-LY (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
854.3
837.2
970.4
836.4
559.2
971.4 693.3
956.3 546.3
375.3
446.2 480.3 509.3 580.2
939.3 809.3
463.3 661.2
753.4 545.4
435.3
409.3 713.2 791.4 887.4 911.3
347.3 629.3 657.3
329.3
296.2 776.4 972.2
Figure S2q. MS
2spectrum of confirmed MC-LR from Hartbeespoort Dam
Hartbeespoort Dam m/z 995, R t 3.89 min MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
967.7 978.7
599.5
553.5
866.7 571.5
979.7 923.7
552.5
865.6
951.7 600.5
910.6
470.5 682.6 710.5
728.6 867.7
375.3 783.6 838.6
525.5 397.4 453.5
738.6
285.3 303.4 347.4 428.4 510.5 625.5 637.6 784.7
980.3
Hartbeespoort Dam m/z 995, R t 3.89 min MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
967.7 978.7
599.5
553.5
866.7 571.5
979.7 923.7
552.5
865.6
951.7 600.5
910.6
470.5 682.6 710.5
728.6 867.7
375.3 783.6 838.6
525.5 397.4 453.5
738.6
285.3 303.4 347.4 428.4 510.5 625.5 637.6 784.7
980.3
Figure S2r. MS
2spectrum of confirmed MC-LR from M. aeruginosa AB2011/53
AB2011/53
m/z 995, R t 3.89 min MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
978.7 967.7
599.5
553.5
866.7
979.7 571.6
552.5 923.7
865.7 600.5
844.6 951.7
910.6 710.5
470.5 682.6 728.5 838.7 867.7
469.5
883.7 783.6
375.4 397.4
453.5 525.5
625.6
510.5 637.5 729.6
285.4 303.4 347.4 399.4 784.6
980.3
AB2011/53
m/z 995, R t 3.89 min MC-LR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
978.7 967.7
599.5
553.5
866.7
979.7 571.6
552.5 923.7
865.7 600.5
844.6 951.7
910.6 710.5
470.5 682.6 728.5 838.7 867.7
469.5
883.7 783.6
375.4 397.4
453.5 525.5
625.6
510.5 637.5 729.6
285.4 303.4 347.4 399.4 784.6
980.3
Figure S2s. MS
2spectrum of confirmed MC-LY from Hartbeespoort Dam
Hartbeespoort Dam m/z 1002, R t 8.48 min MC-LY
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
868.4
984.6
850.5
494.4 559.3
560.3
970.4 693.4
985.5
446.3 580.3
375.3 509.4
477.4 767.5 817.4 952.5
476.4 901.5
423.3 661.3 699.5 833.5
548.3
397.3 706.4 772.4
310.2 347.3 596.4 643.4 727.4 869.5 939.5 1001.8
Hartbeespoort Dam m/z 1002, R t 8.48 min MC-LY
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
868.4
984.6
850.5
494.4 559.3
560.3
970.4 693.4
985.5
446.3 580.3
375.3 509.4
477.4 767.5 817.4 952.5
476.4 901.5
423.3 661.3 699.5 833.5
548.3
397.3 706.4 772.4
310.2 347.3 596.4 643.4 727.4 869.5 939.5 1001.8
Figure S2t. MS
2spectrum of tentative MC-HilR from Hartbeespoort Dam
Hartbeespoort Dam m/z 1009, R t 4.15 min MC-HilR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
990.4 981.5
992.5 599.4
567.4
880.5 566.4
937.6 993.5
571.4 879.6
858.4 965.5
600.4 924.5
728.4
682.4 852.5
881.5 484.4
549.4
375.3 897.6
469.4 739.4 797.5
396.3 539.4 602.4
446.3 625.3 798.5
285.3 347.3
995.4
Hartbeespoort Dam m/z 1009, R t 4.15 min MC-HilR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
990.4 981.5
992.5 599.4
567.4
880.5 566.4
937.6 993.5
571.4 879.6
858.4 965.5
600.4 924.5
728.4
682.4 852.5
881.5 484.4
549.4
375.3 897.6
469.4 739.4 797.5
396.3 539.4 602.4
446.3 625.3 798.5
285.3 347.3
995.4
Figure S2u. MS
2spectrum of tentative MC-HilR from M. aeruginosa AB2011/53
AB2011/53
m/z 1009, R t 4.15 min MC-HilR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
992.7 981.7
599.5
567.5
993.7 880.7
566.5
571.5 879.7 937.7
858.6 924.7 965.7 600.5
710.5
881.7
728.5 852.7
484.4 682.6
375.3 797.6 882.7
539.4
469.5 602.5 739.6 752.5 840.5
446.3 651.5
396.5
347.4 501.4
285.3 428.3
AB2011/53
m/z 1009, R t 4.15 min MC-HilR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
992.7 981.7
599.5
567.5
993.7 880.7
566.5
571.5 879.7 937.7
858.6 924.7 965.7 600.5
710.5
881.7
728.5 852.7
484.4 682.6
375.3 797.6 882.7
539.4
469.5 602.5 739.6 752.5 840.5
446.3 651.5
396.5
347.4 501.4
285.3 428.3
Figure S2v. MS
2spectrum of tentative [Mser
7]MC-LR from Hartbeespoort Dam
Hartbeespoort Dam m/z 1013, R t 3.65 min [Mser 7 ]MC-LR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
995.4
996.5
985.5 599.4
571.4
810.4
969.5 884.5
570.4 941.5 997.5
393.3 453.4 469.5 528.4 600.5 682.4 710.4 728.5 753.5 783.5 844.4 862.4 375.2
303.3 338.5 1000.5
Hartbeespoort Dam m/z 1013, R t 3.65 min [Mser 7 ]MC-LR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
995.4
996.5
985.5 599.4
571.4
810.4
969.5 884.5
570.4 941.5 997.5
393.3 453.4 469.5 528.4 600.5 682.4 710.4 728.5 753.5 783.5 844.4 862.4 375.2
303.3 338.5 1000.5
Figure S2w. MS
2spectrum of tentative [Mser
7]MC-LR from M. aeruginosa AB2011/53
AB2011/53
m/z 1013, R t 3.65 min [Mser 7 ]MC-LR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
995.5
996.5
571.4
599.4 985.5
976.4
997.5 884.5
941.5
553.5 630.6 689.2 728.6 922.0
393.4
809.9
470.3 508.4 743.5 783.5 837.6
453.3 375.3
365.2 416.4 682.8 693.4
300.1 339.9 999.6
AB2011/53
m/z 1013, R t 3.65 min [Mser 7 ]MC-LR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
995.5
996.5
571.4
599.4 985.5
976.4
997.5 884.5
941.5
553.5 630.6 689.2 728.6 922.0
393.4
809.9
470.3 508.4 743.5 783.5 837.6
453.3 375.3
365.2 416.4 682.8 693.4
300.1 339.9 999.6
Figure S2x. MS
2spectrum of unidentified MC-XR from Hartbeespoort Dam
Hartbeespoort Dam m/z 1013, R t 3.77 min Unidentified MC-XR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
985.5
996.4
571.4 599.4
570.3
984.6 941.5 884.5
600.5 997.5
928.4 968.4 879.5
488.4
469.4 569.3 682.5 885.5
728.4 543.3
375.3 801.5 856.4
912.5
606.4 655.4 729.4
509.4 756.1
400.2 446.2 849.3
285.4 339.3 357.3 998.6
Hartbeespoort Dam m/z 1013, R t 3.77 min Unidentified MC-XR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
985.5
996.4
571.4 599.4
570.3
984.6 941.5 884.5
600.5 997.5
928.4 968.4 879.5
488.4
469.4 569.3 682.5 885.5
728.4 543.3
375.3 801.5 856.4
912.5
606.4 655.4 729.4
509.4 756.1
400.2 446.2 849.3
285.4 339.3 357.3 998.6
Figure S2y. MS
2spectrum of unidentified MC-XR from M. aeruginosa AB2011/53
AB2011/53
m/z 1013, R t 3.77 min Unidentified MC-XR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
995.5 985.5
996.5
571.3
599.4
884.5
997.5 941.5
570.4 883.5
964.5
600.5 879.4 928.4
488.3 682.5 862.4 901.5
470.4
375.3 728.5
554.3
528.5 683.4 856.4
469.5 625.3 756.2 834.5
417.2 654.5
347.4 285.4
AB2011/53
m/z 1013, R t 3.77 min Unidentified MC-XR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
995.5 985.5
996.5
571.3
599.4
884.5
997.5 941.5
570.4 883.5
964.5
600.5 879.4 928.4
488.3 682.5 862.4 901.5
470.4
375.3 728.5
554.3
528.5 683.4 856.4
469.5 625.3 756.2 834.5
417.2 654.5
347.4 285.4
Figure S2z. MS
2spectrum of tentative [Asp
3]MC-RR from Hartbeespoort Dam
Hartbeespoort Dam m/z 1024, R t 2.12 min [Asp 3 ]MC-RR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
965.7
1006.6
966.7
895.6
1007.7
967.8 982.7 962.7 728.6
599.5
892.5
944.7 896.7
616.6 779.5
581.5 710.7 745.6 797.6 875.6
426.6 531.4 675.5
340.4 460.5 521.6 1008.7
314.4 411.5
Hartbeespoort Dam m/z 1024, R t 2.12 min [Asp 3 ]MC-RR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
965.7
1006.6
966.7
895.6
1007.7
967.8 982.7 962.7 728.6
599.5
892.5
944.7 896.7
616.6 779.5
581.5 710.7 745.6 797.6 875.6
426.6 531.4 675.5
340.4 460.5 521.6 1008.7
314.4 411.5
Figure S2aa. MS
2spectrum of unidentified possible MC-RR analogue from Hartbeespoort Dam
Hartbeespoort Dam m/z 1029, R t 3.37 min
Unidentified MC-RR analogue?
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
965.5
966.5
1011.4
599.4 803.5 836.4 900.5 937.5 947.5
523.4 540.3 682.4 710.4 759.5 967.4
504.4 440.4
416.3
375.3 606.5
352.2 329.4
Hartbeespoort Dam m/z 1029, R t 3.37 min
Unidentified MC-RR analogue?
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
965.5
966.5
1011.4
599.4 803.5 836.4 900.5 937.5 947.5
523.4 540.3 682.4 710.4 759.5 967.4
504.4 440.4
416.3
375.3 606.5
352.2 329.4
Figure S2ab. MS
2spectrum of tentative MC-FR from Hartbeespoort Dam
Hartbeespoort Dam m/z 1029, R t 4.21 min MC-FR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1001.6
1012.6
599.5
587.5
586.4 900.7 957.6
604.5
899.6
571.5 944.6 980.6
878.5
710.5 872.6
682.5
504.4 728.6 901.6
375.3 416.3 469.5 559.4 622.5 759.4 817.6
544.4 431.3
373.3 397.5 671.5 816.6 850.6
303.4
Hartbeespoort Dam m/z 1029, R t 4.21 min MC-FR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1001.6
1012.6
599.5
587.5
586.4 900.7 957.6
604.5
899.6
571.5 944.6 980.6
878.5
710.5 872.6
682.5
504.4 728.6 901.6
375.3 416.3 469.5 559.4 622.5 759.4 817.6
544.4 431.3
373.3 397.5 671.5 816.6 850.6
303.4
Figure S2ac. MS
2spectrum of tentative MC-FR from M. aeruginosa AB2011/53
AB2011/53
m/z 1029, R t 4.21 min MC-FR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1001.6
1012.6
599.5
587.4
586.5 900.6
957.7
604.5 899.6
944.6 878.6
571.5 980.6
682.5 710.5
504.4 728.6 817.6 872.6 901.7
759.5 416.3
375.4397.4 433.4 469.5 544.5 559.5 622.4 671.5 835.6
373.3 799.5
303.4
AB2011/53
m/z 1029, R t 4.21 min MC-FR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1001.6
1012.6
599.5
587.4
586.5 900.6
957.7
604.5 899.6
944.6 878.6
571.5 980.6
682.5 710.5
504.4 728.6 817.6 872.6 901.7
759.5 416.3
375.4397.4 433.4 469.5 544.5 559.5 622.4 671.5 835.6
373.3 799.5
303.4
Figure S2ad. MS
2spectrum of tentative MC-RF from Hartbeespoort Dam
Hartbeespoort Dam m/z 1029, R t 6.34 min MC-RF (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1011.4
440.4 912.4
1001.5 865.5
957.5 950.4
753.5 913.4
984.6
586.4 868.5
329.3
754.3 731.4
587.3 441.3
780.5
721.3 803.4 937.6
605.4
422.9 461.3 509.2 552.3 667.2 704.5 827.5
371.2 417.2 327.2
Hartbeespoort Dam m/z 1029, R t 6.34 min MC-RF (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1011.4
440.4 912.4
1001.5 865.5
957.5 950.4
753.5 913.4
984.6
586.4 868.5
329.3
754.3 731.4
587.3 441.3
780.5
721.3 803.4 937.6
605.4
422.9 461.3 509.2 552.3 667.2 704.5 827.5
371.2 417.2 327.2
Figure S2ae. MS
2spectrum of tentative [DMAdda
5]MC-YR from Hartbeespoort Dam
Hartbeespoort Dam m/z 1031, R t 2.40 min
[DMAdda 5 ]MC-YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1003.5 1013.5
1014.5
585.4
603.4
894.4
959.5 911.4 557.4
620.4
1015.4 987.5
946.4 912.5 668.4 696.4
520.3 714.4 819.5 874.5
432.3 455.4 638.4 788.4 866.4 920.5
375.3 431.2 457.3 519.3 537.3 775.4
371.3
303.3 1016.4
Hartbeespoort Dam m/z 1031, R t 2.40 min
[DMAdda 5 ]MC-YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1003.5 1013.5
1014.5
585.4
603.4
894.4
959.5 911.4 557.4
620.4
1015.4 987.5
946.4 912.5 668.4 696.4
520.3 714.4 819.5 874.5
432.3 455.4 638.4 788.4 866.4 920.5
375.3 431.2 457.3 519.3 537.3 775.4
371.3
303.3 1016.4
Figure S2af. MS
2spectrum of tentative [DMAdda
5]MC-YR from M. aeruginosa AB2011/53
AB2011/53
m/z 1031, R t 2.40 min
[DMAdda 5 ]MC-YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1003.6
1013.6
585.5
603.4
894.5 911.6
959.7
620.5 1015.7
557.5
946.6
986.7
520.5 668.5 696.6 714.6 874.6 912.6
638.5
919.7
375.4 431.5 455.5 519.5 537.4 769.6 788.5 819.6
845.7
347.4 456.5 758.8
309.9 422.3 1026.4
AB2011/53
m/z 1031, R t 2.40 min
[DMAdda 5 ]MC-YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1003.6
1013.6
585.5
603.4
894.5 911.6
959.7
620.5 1015.7
557.5
946.6
986.7
520.5 668.5 696.6 714.6 874.6 912.6
638.5
919.7
375.4 431.5 455.5 519.5 537.4 769.6 788.5 819.6
845.7
347.4 456.5 758.8
309.9 422.3 1026.4
Figure S2ag. MS
2spectrum of tentative [Asp
3]MC-YR from Hartbeespoort Dam
Hartbeespoort Dam m/z 1031, R t 3.47 min [Asp 3 ]MC-YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1003.5
1014.5
599.4
589.4
959.5 588.4
571.4 606.4
1015.5 982.4
901.5 880.4
916.5 375.3
934.5
696.4 761.4 819.5
668.5 714.4
469.4 488.4 546.3 624.3 744.3 774.4 868.4
433.2 655.4
397.4 357.3
289.3 320.2 1017.6
Hartbeespoort Dam m/z 1031, R t 3.47 min [Asp 3 ]MC-YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1003.5
1014.5
599.4
589.4
959.5 588.4
571.4 606.4
1015.5 982.4
901.5 880.4
916.5 375.3
934.5
696.4 761.4 819.5
668.5 714.4
469.4 488.4 546.3 624.3 744.3 774.4 868.4
433.2 655.4
397.4 357.3
289.3 320.2 1017.6
Figure S2ah. MS
2spectrum of tentative [Asp
3]MC-YR from M. aeruginosa AB2011/53
AB2011/53
m/z 1031, R t 3.47 min [Asp 3 ]MC-YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1013.5
771.6 420.3
1003.6 1014.6
599.5
787.4 846.3
589.5 769.4 789.5 959.7
571.5 606.5 880.5 901.7 982.6 1015.7
537.1 696.6 916.7
375.4 459.4 469.6 527.4 625.5 668.6 744.6 805.4 879.7
310.3 348.4 1031.9
AB2011/53
m/z 1031, R t 3.47 min [Asp 3 ]MC-YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1013.5
771.6 420.3
1003.6 1014.6
599.5
787.4 846.3
589.5 769.4 789.5 959.7
571.5 606.5 880.5 901.7 982.6 1015.7
537.1 696.6 916.7
375.4 459.4 469.6 527.4 625.5 668.6 744.6 805.4 879.7
310.3 348.4 1031.9
Figure S2ai. MS
2spectrum of tentative [Dha
7]MC-YR from Hartbeespoort Dam
Hartbeespoort Dam m/z 1031, R t 3.75 min [Dha 7 ]MC-YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
599.4
1003.5
589.4
1014.5
588.3 902.5
571.4
901.5 606.4
880.4 1015.5
710.4
959.5 728.5
903.5
554.5 625.5 682.4
668.5 913.5
546.4
432.3 469.6 875.5
333.3 361.1 387.4 444.4 492.2 520.4 632.4 774.5 846.5
303.4 1016.4
Hartbeespoort Dam m/z 1031, R t 3.75 min [Dha 7 ]MC-YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
599.4
1003.5
589.4
1014.5
588.3 902.5
571.4
901.5 606.4
880.4 1015.5
710.4
959.5 728.5
903.5
554.5 625.5 682.4
668.5 913.5
546.4
432.3 469.6 875.5
333.3 361.1 387.4 444.4 492.2 520.4 632.4 774.5 846.5
303.4 1016.4
Figure S2aj. MS
2spectrum of tentative [Dha
7]MC-YR from M. aeruginosa AB2011/53
AB2011/53
m/z 1031, R t 3.75 min [Dha 7 ]MC-YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1003.7 599.5
1013.6 1014.6
589.5
588.5
902.7
571.5
600.5
917.7
901.7 946.7 959.7 1015.6 897.6
710.7 555.2
711.6 700.4 668.5 470.5
361.5 414.6 433.3 502.2 539.4 628.7 746.3 797.9 869.7
319.5
1030.8
AB2011/53
m/z 1031, R t 3.75 min [Dha 7 ]MC-YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1003.7 599.5
1013.6 1014.6
589.5
588.5
902.7
571.5
600.5
917.7
901.7 946.7 959.7 1015.6 897.6
710.7 555.2
711.6 700.4 668.5 470.5
361.5 414.6 433.3 502.2 539.4 628.7 746.3 797.9 869.7
319.5
1030.8
Figure S2ak. MS
2spectrum of tentative [Asp
3]MC-RY from Hartbeespoort Dam
Hartbeespoort Dam m/z 1031, R t 5.12 min [Asp 3 ]MC-RY (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1003.6 426.4
959.6
851.5
1013.6 588.4
1014.6 916.6
960.7 606.5
897.6 754.5
717.5
986.6
736.5 852.6
581.4 329.4 354.4 427.4
917.7 873.6
624.5
311.3 375.4 409.3 492.5 505.5 537.4555.4 673.5 704.5 761.5 819.6 846.6 934.7 1015.6
645.6
1031.4
Hartbeespoort Dam m/z 1031, R t 5.12 min [Asp 3 ]MC-RY (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1003.6 426.4
959.6
851.5
1013.6 588.4
1014.6 916.6
960.7 606.5
897.6 754.5
717.5
986.6
736.5 852.6
581.4 329.4 354.4 427.4
917.7 873.6
624.5
311.3 375.4 409.3 492.5 505.5 537.4555.4 673.5 704.5 761.5 819.6 846.6 934.7 1015.6
645.6
1031.4
Figure S2al. MS
2spectrum of tentative trace [Asp
3]MC-RY from M. aeruginosa AB2011/53
AB2011/53
m/z 1031, R t 5.12 min
[Asp 3 ]MC-RY (tentative trace?)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1003.5
968.4
1013.9 984.8
898.5
851.2 963.5
588.4 930.7
426.3
581.6
609.9 783.7
933.5
427.2 862.5
735.6 743.2
338.1 384.1 462.3 486.7 537.1 647.1 694.5717.5 808.8 829.8 871.1893.9
565.4 506.4
374.3
436.6 385.5
305.2 350.3 1015.5
308.2 1017.9
AB2011/53
m/z 1031, R t 5.12 min
[Asp 3 ]MC-RY (tentative trace?)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1003.5
968.4
1013.9 984.8
898.5
851.2 963.5
588.4 930.7
426.3
581.6
609.9 783.7
933.5
427.2 862.5
735.6 743.2
338.1 384.1 462.3 486.7 537.1 647.1 694.5717.5 808.8 829.8 871.1893.9
565.4 506.4
374.3
436.6 385.5
305.2 350.3 1015.5
308.2 1017.9
Figure S2am. MS
2spectrum of tentative [DMAdda
5]C-(H
4)YR from Hartbeespoort Dam
AB2011/53
m/z 1035, R t 2.06 min
[DMAdda 5 ]MC-(H 4 )YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1017.6
1018.6 1007.7
585.5
973.8 915.7
963.8 999.3 589.4 624.5
869.7 906.6 558.5
1019.6 950.6
853.7
418.3 435.5 506.6 529.4 668.5 721.9 771.7 792.4
401.6 333.3
303.2
AB2011/53
m/z 1035, R t 2.06 min
[DMAdda 5 ]MC-(H 4 )YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1017.6
1018.6 1007.7
585.5
973.8 915.7
963.8 999.3 589.4 624.5
869.7 906.6 558.5
1019.6 950.6
853.7
418.3 435.5 506.6 529.4 668.5 721.9 771.7 792.4
401.6 333.3
303.2
Figure S2an. MS
2spectrum of tentative [Asp
3]MC-(H
4)YR from Hartbeespoort Dam
Hartbeespoort Dam m/z 1035, R t 3.25 min
[Asp 3 ]MC-(H 4 )YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1017.6
1018.6
1007.6
599.5
963.6
593.5 610.5 905.6
575.5 696.5 714.5 866.6 884.6 920.6 1019.6
375.3 421.4 469.5 492.4 509.4 558.4 668.6 765.5 823.6
347.3 326.3
Hartbeespoort Dam m/z 1035, R t 3.25 min
[Asp 3 ]MC-(H 4 )YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1017.6
1018.6
1007.6
599.5
963.6
593.5 610.5 905.6
575.5 696.5 714.5 866.6 884.6 920.6 1019.6
375.3 421.4 469.5 492.4 509.4 558.4 668.6 765.5 823.6
347.3 326.3
Figure S2ao. MS
2spectrum of tentative [Asp
3]MC-(H
4)YR from M. aeruginosa AB2011/53
AB2011/53
m/z 1035, R t 3.25 min
[Asp 3 ]MC-(H 4 )YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1017.6
1018.7
1007.6
599.5
973.7 585.5
571.6 906.6 963.6
610.6 884.6
510.4 624.5 792.6 937.4 1019.6
489.3 689.7 714.7 775.6
379.4 431.4 550.5 824.8
357.6 862.4
338.8
AB2011/53
m/z 1035, R t 3.25 min
[Asp 3 ]MC-(H 4 )YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1017.6
1018.7
1007.6
599.5
973.7 585.5
571.6 906.6 963.6
610.6 884.6
510.4 624.5 792.6 937.4 1019.6
489.3 689.7 714.7 775.6
379.4 431.4 550.5 824.8
357.6 862.4
338.8
Figure S2ap. MS
2spectrum of tentative [Dha
7]MC-(H
4)YR from Hartbeespoort Dam
Hartbeespoort Dam m/z 1035, R t 3.52 min
[Dha 7 ]MC-(H 4 )YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1017.6
1018.6 599.5
1007.7
592.6
906.6
971.6 610.5
571.5
963.7 905.6
682.5 710.5 878.7 907.6
418.5 469.7 531.5 547.5 626.5 668.4 728.6 765.6 805.6 866.6 1019.6
400.4 325.4 369.1
Hartbeespoort Dam m/z 1035, R t 3.52 min
[Dha 7 ]MC-(H 4 )YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1017.6
1018.6 599.5
1007.7
592.6
906.6
971.6 610.5
571.5
963.7 905.6
682.5 710.5 878.7 907.6
418.5 469.7 531.5 547.5 626.5 668.4 728.6 765.6 805.6 866.6 1019.6
400.4 325.4 369.1
Figure S2aq. MS
2spectrum of tentative [Dha
7]MC-(H
4)YR from M. aeruginosa AB2011/53
AB2011/53
m/z 1035, R t 3.52 min
[Dha 7 ]MC-(H 4 )YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1017.6
1018.6
1007.8 599.5
999.8 906.8
950.1 992.6 610.5
592.4
905.9 935.5 1019.6
575.6 435.3
722.4 806.9 839.5
523.4 565.5 611.5 780.8
421.3 470.6 646.4 686.3 868.8
309.7 358.2 390.2
AB2011/53
m/z 1035, R t 3.52 min
[Dha 7 ]MC-(H 4 )YR (tentative)
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1017.6
1018.6
1007.8 599.5
999.8 906.8
950.1 992.6 610.5
592.4
905.9 935.5 1019.6
575.6 435.3
722.4 806.9 839.5
523.4 565.5 611.5 780.8
421.3 470.6 646.4 686.3 868.8
309.7 358.2 390.2
Figure S2ar. MS
2spectrum of tentative MC-AnaR from M. aeruginosa AB2011/53
AB2011/53
m/z 1037, R t 5.05 min MC-AnaR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1009.7
1019.7
599.5
595.5
965.8 594.5
908.7
571.5 612.5 886.6
975.8 952.7 880.7
710.5 682.6 728.6
512.5 925.8
375.3397.4 424.4 469.5 529.5567.5 630.5 679.5 750.6767.6780.6 825.7 868.6 347.4
303.3
AB2011/53
m/z 1037, R t 5.05 min MC-AnaR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1009.7
1019.7
599.5
595.5
965.8 594.5
908.7
571.5 612.5 886.6
975.8 952.7 880.7
710.5 682.6 728.6
512.5 925.8
375.3397.4 424.4 469.5 529.5567.5 630.5 679.5 750.6767.6780.6 825.7 868.6 347.4
303.3
Figure S2as. MS
2spectrum of confirmed MC-RR from Hartbeespoort Dam
Hartbeespoort Dam m/z 1038, R t 2.13 min MC-RR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1020.7
996.7
909.7 978.7
867.7
728.6 926.7 958.7
599.6 710.5 753.6 770.6 822.6
440.4 535.5 571.6 625.5 682.6
375.4 488.5
303.4 328.5 424.6
Hartbeespoort Dam m/z 1038, R t 2.13 min MC-RR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1020.7
996.7
909.7 978.7
867.7
728.6 926.7 958.7
599.6 710.5 753.6 770.6 822.6
440.4 535.5 571.6 625.5 682.6
375.4 488.5
303.4 328.5 424.6
Figure S2at. MS
2spectrum of confirmed MC-RR from M. aeruginosa AB2011/53
AB2011/53
m/z 1038, R t 2.13 min MC-RR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1020.5
996.5 909.5
867.5 978.5
728.4 910.5 976.5
599.4 710.4 753.5 770.5 822.5 866.6 440.3
303.2 347.3 375.3 397.6 457.5 488.0 535.4 569.4 625.4 681.5
AB2011/53
m/z 1038, R t 2.13 min MC-RR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1020.5
996.5 909.5
867.5 978.5
728.4 910.5 976.5
599.4 710.4 753.5 770.5 822.5 866.6 440.3
303.2 347.3 375.3 397.6 457.5 488.0 535.4 569.4 625.4 681.5
Figure S2au. MS
2spectrum of confirmed MC-YR from Hartbeespoort Dam
Hartbeespoort Dam m/z 1045, R t 3.84 min MC-YR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000 1050
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1017.5
1028.4
599.4
603.3
916.5
973.5 620.4
915.5 571.4
960.4
894.3 996.4
682.4 710.4 917.5
520.3 728.4 833.5 888.5
375.3397.4 432.3 469.4 519.4 560.4 638.4639.4 775.3 788.3 882.5 933.5 347.3
303.3
Hartbeespoort Dam m/z 1045, R t 3.84 min MC-YR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000 1050
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1017.5
1028.4
599.4
603.3
916.5
973.5 620.4
915.5 571.4
960.4
894.3 996.4
682.4 710.4 917.5
520.3 728.4 833.5 888.5
375.3397.4 432.3 469.4 519.4 560.4 638.4639.4 775.3 788.3 882.5 933.5 347.3
303.3
Figure S2av. MS
2spectrum of confirmed MC-YR from M. aeruginosa AB2011/53
AB2011/53
m/z 1045, R t 3.84 min MC-YR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000 1050
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1017.6
1028.6
599.5
603.4
973.6
620.5 916.6
571.5 894.6 960.6
983.6
682.5 710.5 888.7 917.6
520.4 728.5
833.6
375.3397.4 432.3 469.5488.5519.5 560.4 638.4639.5 775.5 788.5 882.7 347.4
303.3
AB2011/53
m/z 1045, R t 3.84 min MC-YR
300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000 1050
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Relative Abundance
1017.6
1028.6
599.5
603.4
973.6
620.5 916.6
571.5 894.6 960.6
983.6
682.5 710.5 888.7 917.6
520.4 728.5
833.6
375.3397.4 432.3 469.5488.5519.5 560.4 638.4639.5 775.5 788.5 882.7 347.4
303.3
