April 3, 2008 · Web viewTitle: Assessing impact of algal species indigenous to oil sands process-affected waters on mixtures of oil sands naphthenic acids Authors: Sarah E. Ruffella,

Title: Assessing impact of algal species indigenous to oil sands process-affected waters on mixtures of oil sands naphthenic acids

Authors:

Sarah E. Ruffella, Richard A. Franka, Adam P. Woodwortha, Anthony E. Bauera, Leslie M. Bragga, Lorna E. Deethb, Kirsten M.

Müllera, Andrea J. Farwella, D. George Dixona, Mark R. Servosa, and Brendan J. McConkeya*

aDepartment of Biology, University of Waterloo, Waterloo, ON, Canada N2L 3G1

bDepartment of Mathematics & Statistics, University of Guelph, Guelph, ON, Canada

N1G 2W1

*Corresponding Author: Brendan J. McConkey

Department of Biology

University of Waterloo

Waterloo, ON, Canada N2L 3G1

Phone: (519)888-4567 x37020

Fax: (519)746-0614

Email: [email protected]

mailto:[email protected]

SUPPORTING INFORMATION:

Table S1: Slopes of linear regressions of "all ion" and "marker ion" data at each test concentration (Lower, Upper 95% CI)

Stichococcus sp. 1 Scenedesmus sp. 1All ions Marker ions All ions Marker ions

No. of characterized ions 209 9 209 17

Control 11000* (5570, 16400) 75.3 (-18.9, 170) 4630 (2900, 6360) -106 (-409, 197)p-value 0.00489 0.0907 0.00176 0.385

10 mg L-1 1970 (-4000, 7940) -2160* (-3810, -509) 7600 (-3230, 18400) 1940 (-4750, 8630)p-value 0.412 0.0221 0.123 0.465

30 mg L-1 -1830 (-4660, 997) -1960* (-3290, -623) 8500 (343, 16700) 3700 (-6270, 13700)p-value 0.147 0.0152 0.0444 0.361

100 mg L-1-7160 (-20300, 6020)

-4080* (-7707.456, -456.544) 1800 (-5020, 8620) -1650 (-5450, 2160)

p-value 0.206 0.0353 0.505 0.296Oocystis sp. 1 Chlorella sp. 1

All ions Marker ions All ions Marker ionsNo. of

characterized ions 209 5 209 5Control 2600 (-1140, 6340) 11.6 (-59.7, 82.8) 2380 (1150, 3610) n/ap-value 0.126 0.676 0.00587 n/a

10 mg L-1 6040 (-16200, 28300) 350 (-2660, 3360) 956 (-4050, 5960) -15.3 (-705, 675)p-value 0.493 0.763 0.624 0.954

30 mg L-1 16100 (2170, 30000) 331 (-271, 933) -2010 (-7670, 3650) -127 (-930, 676)p-value 0.0326 0.201 0.380 0.684

100 mg L-1 -4140 (-19300, 11000) -1200 (-3830, 1400) 269 (-4130, 4670) -114 (-919, 691)p-value 0.490 0.275 0.873 0.714

Notes: " * " denotes slope significantly different from 0 (α = 0.05); " n/a " indicates insufficient marker ion data for regression analysis

Table S1 (cont.): Slopes of linear regressions of "all ion" and "marker ion" data at each test concentration (Lower, Upper 95% CI)Aphanocapsa sp. 1 Euglena sp. 1


characterized ions 209 6 209 6Control 5750 (1930, 9570) -9.08 (-43.3, 25.2) 6701* (3700, 9700) 0 (n/a)p-value 0.0139 0.503 0.00344 n/a

10 mg L-1 3260 (510, 6010) 131 (-401, 663) 4150* (1920, 6390) 67.9 (-373, 509)p-value 0.0302 0.533 0.00672 0.691

30 mg L-1 -1150 (-5370, 3070) -118 (-471, 236) 2440 (-177, 5060) -29.5 (-300, 241)p-value 0.492 0.407 0.0608 0.777

100 mg L-1 -2330 (-5430, 758) -107 (-598, 384) -182 (-3070, 2710) 38.7 (-192, 270)p-value 0.104 0.577 0.870 0.666

Anabaena sp. 1 Monoraphidium sp. 1All ions Marker ions All ions Marker ions


Control 381 (-122, 883) n/a 3380 (3020, 3740) n/ap-value 0.103 n/a 0.0000130 n/a

10 mg L-1 -1210 (-4480, 2070) -362 (-1310, 585) 3094 (361.4, 5830) 312 (-405, 1030)p-value 0.364 0.349 0.0347 0.294

30 mg L-1 -231 (-2630, 2170) -20.7 (-195, 153) 3540 (-359, 7440) 1480 (141, 2820)p-value 0.803 0.758 0.0653 0.0374

100 mg L-1 2110 (-846, 5070) -62.1 (-505, 381) 1340 (382, 2300) 152 (-973, 1280)p-value 0.119 0.717 0.0179 0.727


Table S1 (cont.): Slopes of linear regressions of "all ion" and "marker ion" data at each test concentration (Lower, Upper 95% CI)Scenedesmus sp. 3 Monoraphidium sp. 2


characterized ions 209 21 209 22Control -864 (-2930, 1200) -44.7 (-171, 81.2) 2040 (752, 3330) 3.96 (-138, 146)p-value 0.310 0.381 0.0118 0.942

10 mg L-1 -740 (-4420, 2940) -175 (-1860, 1510) 2190 (-2020, 6390) -1070 (-2690, 550)p-value 0.607 0.787 0.222 0.141

30 mg L-1 -346 (-4610, 3920) -134 (-3260, 2990) 3840 (79.7, 7610) 91.5 (-2020, 2200)p-value 0.833 0.910 0.0471 0.910

100 mg L-1 1110 (-6330, 8540) -22.5 (-3280, 3240) 9020 (-7200, 25000) 4520 (-8300, 17300)p-value 0.700 0.986 0.197 0.383

Oocystis sp. 2 Unidentified sp. 1All ions Marker ions All ions Marker ions


Control 159 (-2190, 2510) -47.1 (-260, 166) 3890 (2280, 5500) -16.2 (-104, 71.4)p-value 0.860 0.573 0.00255 0.635

10 mg L-1 -567 (-14000, 12900) -2040 (-10300, 6210) -127 (-11700, 11400) 368 (-367, 1100)p-value 0.912 0.530 0.977 0.237

30 mg L-1 1390 (-4650, 7420) -288 (-1970, 1390) -1200 (-10950, 8550) -40.4 (-1350, 1270)p-value 0.558 0.659 0.750 0.936

100 mg L-1 1640 (-6280, 9570) 1210 (-2150, 4570) -1120 (-16300, 14000) -325 (-1800, 1150)p-value 0.596 0.374 0.847 0.575


Table S1 (cont.): Slopes of linear regressions of "all ion" and "marker ion" data at each test concentration (Lower, Upper 95% CI)Tetraedron sp. 1 Ulothrix sp. 1


characterized ions 209 21 209 12Control 2380 (586, 4180) 6.67 (-207, 220) 2390 (-1100, 5870) 84.9 (-54.3, 224)p-value 0.0212 0.935 0.130 0.166

10 mg L-1 117 (-21800, 22000) -794 (-10400, 8800) 1050 (-3660, 5770) -466 (-2200, 1270)p-value 0.989 0.830 0.569 0.497

30 mg L-1 7540 (-13200, 28300) 3480 (-8580, 15500) 1220 (-5710, 8150) -240 (-3630, 3160)p-value 0.370 0.468 0.651 0.854

100 mg L-1 2610 (-8460, 13700) 3160 (-7260, 13600) 640 (-9240, 10500) 490 (-1200, 2180)p-value 0.549 0.448 0.866 0.467

Scenedesmus sp. 4 Chlorella sp. 3All ions Marker ions All ions Marker ions


Control 4800 (-1790, 11400) 25.1 (-339, 389) -2330 (-7440, 2790) -704 (-1840, 430)p-value 0.113 0.858 0.275 0.160

10 mg L-1 6130 (-2500, 14800) 1500 (-2700, 5700) -2980 (-10200, 4240) -4700 (-10400, 1020)p-value 0.120 0.376 0.316 0.0846

30 mg L-1 3110 (-4130, 10300) 252 (-2990, 3500) -9330 (-34100, 15400) -5990 (-19600, 7610)p-value 0.299 0.840 0.355 0.288

100 mg L-1 7990* (2220, 13800) -191 (-5470, 5090) 1730 (-8200, 11700) 193 (-6900, 7280)p-value 0.0184 0.925 0.654 0.944


Table S1 (cont.): Slopes of linear regressions of "all ion" and "marker ion" data at each test concentration (Lower, Upper 95% CI)Unidentified sp. 2

All ions Marker ionsNo. of

characterized ions 209 12Control -434 (-2100, 1240) 67.8 (-44.1, 180)p-value 0.510 0.168

10 mg L-1 1770 (-4650, 8190) -82.8 (-25240 2360)p-value 0.487 0.930

30 mg L-1 3660 (-5560, 12900) 1060 (-1960, 4080)p-value 0.332 0.385

100 mg L-1 1220 (-1820, 4250) 325 (-1870, 2520)p-value 0.328 0.703


Figure Captions:

Figure S1: Linearity of the concentration-response from 1 mg L-1 to 75 mg L-1 for a previously characterized naphthenic acid aliquot (Rogers et al., 2002). (A) total ion abundance; (B) marker ion abundance.

Figure S2: Summation of MS peak areas for “All ion” (solid circle) and “Marker ion” (open circle) approaches for 17 algal bioremediation candidate strains. The “All ion” approach characterized and summed the MS peak areas for NAs representing the 209 previously reported “classical NAs” (Lo et al., 2003). The “Marker ion” approach characterized and summed the MS peak areas for the masses identified as being present within the NA extract yet not having interference from biogenic acids released by the phytoplankton strains. Linear regressions were modeled for “All ion” and “Marker ion” approaches using R (R Development Core Team, 2008) with b1 representing the slope of the regression lines.

Figure S1

0 10 20 30 40 50 60 70 800

100000

200000

300000

400000

500000

600000

700000

800000

900000

1000000

f(x) = 11651.5445620159 x + 10390.6651030872R² = 0.999838161399935

A

Concentration, mg L-1

Tota

l Ion

Abu

ndan

ce

0 10 20 30 40 50 60 70 800

50000

100000

150000

200000f(x) = 2622.60259626923 x − 1186.00359986909R² = 0.992918197447135

B

Concentration, mg L-1

Mar

ker I

on A

bund

ance

Figure S2

Sample Day

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

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105

2.0x105

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0 2 4 6 8 10 12 14

MS

Pea

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105

2.0x105

3.0x105

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105

2.0x105

3.0x105

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MS

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k A

rea

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5.0x104

105

1.5x105

2.0x105

2.5x105

100 mg L -1 NA

30 mg L -1 NA

10 mg L -1 NA

Control

b1 = -4081.5

b1 = -7156.1

b1 = -1955.4

b1 = -1831.8

b1 = -2158.2

b1 = 1967.2

b1 = 75.3

b1 = 10987.2

Stichococcus sp.1

Sample Day

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

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105

2.0x105

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Pea

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Pea

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2.0x105

3.0x105

4.0x105

5.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

8.0x104

105

1.2x105

1.4x105

100 mg L -1

30 mg L -1

10 mg L -1

Control

b1 = -

b1 =

b1 =

b1 =

b1 =

b1 =

b1 = -

b1 =

Scenedesmus sp.1

Sample Day

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

105

2.0x105

3.0x105

4.0x105

5.0x105

6.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

105

2.0x105

3.0x105

4.0x105

5.0x105

6.0x105

7.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

105

2.0x105

3.0x105

4.0x105

5.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

5.0x104

105

1.5x105

2.0x105

Control

Oocystis sp. 1

100 mg L -1 NA

30 mg L -1 NA

10 mg L -1 NA

b1 = -1197.5

b1 = -4136.1

b1 = 331.0

b1 = 16062.4

b1 = 350.0

b1 = 6042.5

b1 = 11.6

b1 = 2599.4

Sample Day

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

5.0x104

105

1.5x105

0 2 4 6 8 10 12 14

MS

Pea

k A

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5.0x104

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1.5x105

2.0x105

2.5x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

5.0x104

105

1.5x105

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0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

8.0x104

105

Control

Chlorella sp. 1

100 mg L -1 NA

30 mg L -1 NA

10 mg L -1 NA

b1 = -114.0

b1 = 269.4

b1 = -126.7

b1 = -2012.5

b1 = -15.3

b1 = 955.7

b1 = 0

b1 = 2378.7

Sample Day

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

5.0x104

105

1.5x105

2.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

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5.0x104

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1.5x105

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MS

Pea

k A

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0

5.0x104

105

1.5x105

2.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

8.0x104

105

1.2x105

1.4x105

1.6x105

Control

Aphanocapsa sp. 1

100 mg L -1 NA

30 mg L -1 NA

10 mg L -1 NA

b1 = -107.0

b1 = -2334.0

b1 = -117.8

b1 = -1148.7

b1 = 130.8

b1 = 3259.7

b1 = -9.1

b1 = 5746.9

Sample Day

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

5.0x104

105

1.5x105

2.0x105

0 2 4 6 8 10 12 14

MS

Pea

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rea

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5.0x104

105

1.5x105

2.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

8.0x104

105

1.2x105

1.4x105

1.6x105

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0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

8.0x104

105

1.2x105

1.4x105

1.6x105

Control

Euglena sp. 1

100 mg L -1 NA

30 mg L -1 NA

10 mg L -1 NA

b1 = 38.7

b1 = -181.8

b1 = -29.5

b1 = 2442.9

b1 = 67.9

b1 = 4154.2

b1 = 0

b1 = 6701.0

Sample Day

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

0 2 4 6 8 10 12 14

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Pea

k A

rea

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104

2.0x104

3.0x104

4.0x104

5.0x104

6.0x104

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

8.0x104

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

5.0x103

104

1.5x104

2.0x104

2.5x104

Control

Anabaena sp. 1

100 mg L -1 NA

30 mg L -1 NA

10 mg L -1 NA

b1 = -62.1

b1 = 2110.1

b1 = -20.7

b1 = -230.9

b1 = -361.5

b1 = -1207.2

b1 = 0

b1 = 380.5

Sample Day

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

8.0x104

105

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0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

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1.2x105

1.4x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

8.0x104

105

1.2x105

1.4x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

8.0x104

105

Control

Monoraphidium sp. 1

100 mg L -1 NA

30 mg L -1 NA

10 mg L -1 NA

b1 = 151.6

b1 =1343.2

b1 = 1481.9

b1 = 3541.4

b1 = 311.9

b1 = 3093.5

b1 = 0

b1 = 3376.7

Sample Day

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

105

2.0x105

3.0x105

4.0x105

5.0x105

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MS

Pea

k A

rea

0

105

2.0x105

3.0x105

4.0x105

0.0 2.0 4.0 6.0 8.0 10.0 12.0 14.0

MS

Pea

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rea

0

105

2.0x105

3.0x105

4.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

8.0x104

105

1.2x105 Control

Scenedesmus sp. 3

100 mg L -1

30 mg L -1

10 mg L -1

b1 = -

b1 =

b1 = -

b1 = -

b1 = -

b1 = -

b1 = -

b1 = -

Sample Day

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

105

2.0x105

3.0x105

4.0x105

5.0x105

6.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

105

2.0x105

3.0x105

4.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

105

2.0x105

3.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

8.0x104

105

Control

Monoraphidium sp. 2

100 mg L -1

30 mg L -1

10 mg L -1

b1 =

b1 =

b1 =

b1 =

b1 = -

b1 =

b1 =

b1 =

Sample Day

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

105

2.0x105

3.0x105

4.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

105

2.0x105

3.0x105

4.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

105

2.0x105

3.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

8.0x104Control

Oocystis sp. 2

100 mg L -1 NA

30 mg L -1 NA

10 mg L -1 NA

b1 = 1209.6

b1 = 1643.7

b1 = -287.9

b1 = 1385.5

b1 = -2042.7

b1 = -567.3

b1 = -47.1

b1 = 159.4

Sample Day

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x105

4.0x105

6.0x105

8.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x105

4.0x105

6.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

105

2.0x105

3.0x105

4.0x105

5.0x105

6.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

8.0x104

105

1.2x105

1.4x105

1.6x105

Control

Unidentified sp. 1

100 mg L -1 NA

30 mg L -1 NA

10 mg L -1 NA

b1 = -324.8

b1 = -1120.6

b1 = -40.4

b1 = -1198.7

b1 = 367.5

b1 = -127.1

b1 = -16.2

b1 = 3891.0

Sample Day

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

105

2.0x105

3.0x105

4.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

105

2.0x105

3.0x105

4.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

105

2.0x105

3.0x105

4.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

8.0x104

105 Control

Tetraedron sp. 1

100 mg L -1 NA

30 mg L -1 NA

10 mg L -1 NA

b1 = 3158.9

b1 = 2604.8

b1 = 3478.5

b1 = 7537.2

b1 = -793.9

b1 = 117.9

b1 = 6.7

b1 = 2380.7

Sample Day

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

105

2.0x105

3.0x105

4.0x105

5.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

105

2.0x105

3.0x105

4.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

105

2.0x105

3.0x105

4.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

8.0x104

105

1.2x105

1.4x105

Control

Ulothrix sp. 1

100 mg L -1 NA

30 mg L -1 NA

10 mg L -1 NA

b1 = 489.5

b1 = 640.4

b1 = -239.5

b1 = 1219.1

b1 = -466.2

b1 = 1053.1

b1 = 84.9

b1 = 2387.8

Sample Day

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x105

4.0x105

6.0x105

8.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x105

4.0x105

6.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

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0

2.0x105

4.0x105

6.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

5.0x104

105

1.5x105

2.0x105 Control

Scenedesmus sp. 4

100 mg L -1

30 mg L -1

10 mg L -1

b1 = -

b1 =

b1 =

b1 =

b1 =

b1 =

b1 =

b1 =

Sample Day

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x105

4.0x105

6.0x105

8.0x105

106

1.2x106

1.4x106

1.6x106

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x105

4.0x105

6.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x105

4.0x105

6.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

5.0x104

105

1.5x105

2.0x105 Control

Chlorella sp. 3

100 mg L -1 NA

30 mg L -1 NA

10 mg L -1 NA

b1 = 192.5

b1 = 1730.7

b1 = -5993.8

b1 = -9329.6

b1 = -4697.3

b1 = -2978.5

b1 = -704.2

b1 = -2327.8

Sample Day

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

5.0x104

105

1.5x105

2.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

5.0x104

105

1.5x105

2.0x105

2.5x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

5.0x104

105

1.5x105

2.0x105

0 2 4 6 8 10 12 14

MS

Pea

k A

rea

0

2.0x104

4.0x104

6.0x104

8.0x104

Control

Unidentified sp. 2

100 mg L -1

30 mg L -1

10 mg L -1

b1 = -

b1 =

b1 =

b1 =

b1 = -

b1 =

b1 =

b1 = -

Documents

April 3, 2008 · Web viewTitle: Assessing impact of algal species indigenous to oil sands process-affected waters on mixtures of oil sands naphthenic acids Authors: Sarah E. Ruffella,