. 2021 Jan 18;9(1):53–67. doi: 10.1093/emph/eoaa051

Table 1.

F statistics and P-values for 24-h growth assays

Substance	Range tested	Population	Concentration	Interaction
Iron (II) resistant > controls
Iron (II) FeSO₄	6.2–2500 mg/l	F = 179.1 P = 0.0001	F = 7.08 P = 0.0001	F = 1.26 P = 0.265
Iron (III) Fe₂(SO4)₃	6.2–1750 mg/l	F = 296.4 P = 0.0001	F = 10.5 P = 0.0001	F = 5.5 P = 0.0001
Gallium Ga(NO₃)_3.	6.2–5000 mg/l	F = 886.4 P = 0.0001	F = 69.3 P = 0.0001	F = 2.5 P = 0.006
Silver nitrate AgNO₃	6.2–250 mg/l	F = 1.059 P = 0.307, NS	F = 10.96 P = 0.0001	F = 0.359 P = 0.951
Sodium sulfate NaSO⁴	6.2–5000 mg/l	F = 133.5 P = 0.0001	F = 13.9 P = 0.0001	F = 6.2 P = 0.0001
Ampicillin	6.2–2500 mg/l	F = 10.1 P = 0.0001	F = 27.9 P = 0.0001	F = 20.7 P = 0.0001
Bacitracin	6.2–2500 mg/l	F = 353.9 P = 0.0001	F = 3.8 P = 0.0001	F = 29.3 P = 0.0001
Chloramphenicol	6.2–2500 mg/l	F = 147.3 P = 0.0001	F = 96.6 P = 0.0001	F = 39.2 P = 0.0001
Polymixin-B	6.2–2500 mg/l	F = 10.2 P = 0.0001	F = 113.9 P = 0.0001	F = 3.6 P = 0.0001
Rifampicin	6.2–2500 mg/l	F = 205.1 P = 0.0001	F = 23.2 P = 0.0001	F = 7.6 P = 0.0001
Sulfanilamide	6.2–2500 mg/l	F = 22.5 P = 0.0001	F = 95.5 P = 0.0001	F = 4.9 P = 0.0001
Iron (II) resistant = controls
Tetracycline	6.2–2500 mg/l	F = 0.306 P = 0.581	F = 158.3 P = 0.0001	F = 0.245 P = 0.951
Iron (II) resistant > ancestor
Iron (II) FeSO₄	6.2–5000 mg/l	F = 154.4 P = 0.0001	F = 3.4 P = 0.001	F = 4.8 P = 0.0001
Iron (III) Fe₂(SO4)₃	6.2–5000 mg/l	F = 169.8 P = 0.0001	F = 3.5 P = 0.001	F = 18.07 P = 0.0001
Gallium Ga(NO₃)_3.	6.2–5000 mg/l	F = 1107.3 P = 0.0001	F = 13.0 P = 0.001	F = 17.8 P = 0.0001
Silver nitrate AgNO₃	6.2–5000 mg/l	F = 1.06 P = 0.310, NS	F = 5.58 P = 0.001	F = 2.04 P = 0.109
Sodium sulfate NaSO⁴	6.2–5000 mg/l	F = 136.6 P = 0.0001	F = 14.1 P = 0.0001	F = 6.5 P = 0.0001
Ampicillin	6.2–2500 mg/l	F = 206.8 P = 0.0001	F = 60.4 P = 0.0001	F = 18.2 P = 0.0001
Bacitracin	6.2–2500 mg/l	F = 900.6 P = 0.0001	F = 28.3 P = 0.0001	F = 5.6 P = 0.0001
Chloramphenicol	6.2–2500 mg/l	F = 121.5 P = 0.0001	F = 94.7 P = 0.0001	F = 30.0 P = 0.001
Polymixin-B	6.2–2500 mg/l	F = 53.0 P = 0.0001	F = 112.7 P = 0.0001	F = 80.1 P = 0.0001
Rifampicin	6.2–2500 mg/l	F = 288.4 P = 0.001	F = 22.1 P = 0.001	F = 7.2 P = 0.0001
Sulfanilamide	6.2–2500 mg/l	F = 198.1 P = 0.0001	F = 98.4 P = 0.0001	F = 24.9 P = 0.001
Tetracycline	6.2–2500 mg/l	F = 35.4 P = 0.0001	F = 110.2 P = 0.001	F = 29.3 P = 0.0001
Controls > ancestor
Iron (II) FeSO₄	6.2–1750 mg/l	F = 5.2 P = 0.02	F = 2.3 P = 0.01	F = 2.5 P = 0.01
Iron (III) Fe₂(SO4)₃	6.2–1750 mg/l	F = 6.3 P = 0.01	F = 10.0 P = 0.0001	F = 8.6 P = 0.0001
Gallium Ga(NO₃)₃	6.2–1000 mg/l	F = 363.8 P = 0.0001	F = 76.0 P = 0.0001	F = 100.0 P = 0.0001
Ampicillin	6.2–100 mg/l	F = 69.7 P = 0.0001	F = 13.8 P = 0.0001	F = 5.9 P = 0.0001
Bacitracin	6.2–1000 mg/l	F = 312.0 P = 0.0001	F = 3.3 P = 0.001	F = 30.0 P = 0.0001
Polymixin-B	6.2–1000 mg/l	F = 20.3 P = 0.0001	F = 119.6 P = 0.0001	F = 2.1 P = 0.02
Rifampicin	6.2–1000 mg/l	F = 95.8 P = 0.001	F = 48.3 P = 0.001	F = 13.2 P = 0.0001
Sulfanilamide	6.2–1000 mg/l	F = 95.8 P = 0.0001	F = 38.9 P = 0.0001	F = 5.3 P = 0.0001
Tetracycline	6.2–1000 mg/l	F = 53.0 P = 0.0001	F = 188.0 P = 0.001	F = 46.7 P = 0.0001
Controls = ancestor
Silver nitrate AgNO₃	6.2–5000 mg/l	F = 3.4 P = 0.06	F = 93.7 P = 0.0001	F = 17.8 P = 0.0001
Sodium sulfate NaSO₄	6.2–5000 mg/l	F = 0.0 P = 1.00	F = 3.0 P = 0.001	F = 0.0 P = 1.0000
Chloramphenicol	6.2–100 mg/l	F = 0.59 P = 0.44	F = 48.1 P = 0.0001	F = 2.4 P = 0.01

Substance

Range tested

Population

Concentration

Interaction

Iron (II) resistant > controls

Iron (II) FeSO₄

6.2–2500 mg/l

F = 179.1

P = 0.0001

F = 7.08

P = 0.0001

F = 1.26

P = 0.265

Iron (III) Fe₂(SO4)₃

6.2–1750 mg/l

F = 296.4

P = 0.0001

F = 10.5

P = 0.0001

F = 5.5

P = 0.0001

Gallium Ga(NO₃)_3.

6.2–5000 mg/l

F = 886.4

P = 0.0001

F = 69.3

P = 0.0001

F = 2.5

P = 0.006

Silver nitrate AgNO₃

6.2–250 mg/l

F = 1.059

P = 0.307, NS

F = 10.96

P = 0.0001

F = 0.359

P = 0.951

Sodium sulfate NaSO⁴

6.2–5000 mg/l

F = 133.5

P = 0.0001

F = 13.9

P = 0.0001

F = 6.2

P = 0.0001

Ampicillin

6.2–2500 mg/l

F = 10.1

P = 0.0001

F = 27.9

P = 0.0001

F = 20.7

P = 0.0001

Bacitracin

6.2–2500 mg/l

F = 353.9

P = 0.0001

F = 3.8

P = 0.0001

F = 29.3

P = 0.0001

Chloramphenicol

6.2–2500 mg/l

F = 147.3

P = 0.0001

F = 96.6

P = 0.0001

F = 39.2

P = 0.0001

Polymixin-B

6.2–2500 mg/l

F = 10.2

P = 0.0001

F = 113.9

P = 0.0001

F = 3.6

P = 0.0001

Rifampicin

6.2–2500 mg/l

F = 205.1

P = 0.0001

F = 23.2

P = 0.0001

F = 7.6

P = 0.0001

Sulfanilamide

6.2–2500 mg/l

F = 22.5

P = 0.0001

F = 95.5

P = 0.0001

F = 4.9

P = 0.0001

Iron (II) resistant = controls

Tetracycline

6.2–2500 mg/l

F = 0.306

P = 0.581

F = 158.3

P = 0.0001

F = 0.245

P = 0.951

Iron (II) resistant > ancestor

Iron (II) FeSO₄

6.2–5000 mg/l

F = 154.4

P = 0.0001

F = 3.4

P = 0.001

F = 4.8

P = 0.0001

Iron (III) Fe₂(SO4)₃

6.2–5000 mg/l

F = 169.8

P = 0.0001

F = 3.5

P = 0.001

F = 18.07

P = 0.0001

Gallium Ga(NO₃)_3.

6.2–5000 mg/l

F = 1107.3

P = 0.0001

F = 13.0

P = 0.001

F = 17.8

P = 0.0001

Silver nitrate AgNO₃

6.2–5000 mg/l

F = 1.06

P = 0.310, NS

F = 5.58

P = 0.001

F = 2.04

P = 0.109

Sodium sulfate NaSO⁴

6.2–5000 mg/l

F = 136.6

P = 0.0001

F = 14.1

P = 0.0001

F = 6.5

P = 0.0001

Ampicillin

6.2–2500 mg/l

F = 206.8

P = 0.0001

F = 60.4

P = 0.0001

F = 18.2

P = 0.0001

Bacitracin

6.2–2500 mg/l

F = 900.6

P = 0.0001

F = 28.3

P = 0.0001

F = 5.6

P = 0.0001

Chloramphenicol

6.2–2500 mg/l

F = 121.5

P = 0.0001

F = 94.7

P = 0.0001

F = 30.0

P = 0.001

Polymixin-B

6.2–2500 mg/l

F = 53.0

P = 0.0001

F = 112.7

P = 0.0001

F = 80.1

P = 0.0001

Rifampicin

6.2–2500 mg/l

F = 288.4

P = 0.001

F = 22.1

P = 0.001

F = 7.2

P = 0.0001

Sulfanilamide

6.2–2500 mg/l

F = 198.1

P = 0.0001

F = 98.4

P = 0.0001

F = 24.9

P = 0.001

Tetracycline

6.2–2500 mg/l

F = 35.4

P = 0.0001

F = 110.2

P = 0.001

F = 29.3

P = 0.0001

Controls > ancestor

Iron (II) FeSO₄

6.2–1750 mg/l

F = 5.2

P = 0.02

F = 2.3

P = 0.01

F = 2.5

P = 0.01

Iron (III) Fe₂(SO4)₃

6.2–1750 mg/l

F = 6.3

P = 0.01

F = 10.0

P = 0.0001

F = 8.6

P = 0.0001

Gallium Ga(NO₃)₃

6.2–1000 mg/l

F = 363.8

P = 0.0001

F = 76.0

P = 0.0001

F = 100.0

P = 0.0001

Ampicillin

6.2–100 mg/l

F = 69.7

P = 0.0001

F = 13.8

P = 0.0001

F = 5.9

P = 0.0001

Bacitracin

6.2–1000 mg/l

F = 312.0

P = 0.0001

F = 3.3

P = 0.001

F = 30.0

P = 0.0001

Polymixin-B

6.2–1000 mg/l

F = 20.3

P = 0.0001

F = 119.6

P = 0.0001

F = 2.1

P = 0.02

Rifampicin

6.2–1000 mg/l

F = 95.8

P = 0.001

F = 48.3

P = 0.001

F = 13.2

P = 0.0001

Sulfanilamide

6.2–1000 mg/l

F = 95.8

P = 0.0001

F = 38.9

P = 0.0001

F = 5.3

P = 0.0001

Tetracycline

6.2–1000 mg/l

F = 53.0

P = 0.0001

F = 188.0

P = 0.001

F = 46.7

P = 0.0001

Controls = ancestor

Silver nitrate AgNO₃

6.2–5000 mg/l

F = 3.4

P = 0.06

F = 93.7

P = 0.0001

F = 17.8

P = 0.0001

Sodium sulfate NaSO₄

6.2–5000 mg/l

F = 0.0

P = 1.00

F = 3.0

P = 0.001

F = 0.0

P = 1.0000

Chloramphenicol

6.2–100 mg/l

F = 0.59

P = 0.44

F = 48.1

P = 0.0001

F = 2.4

P = 0.01

Notes: For statistical analysis, ANOVA using a generalized linear model was used to evaluate the treatment effect associated with iron (II) selection of the 24-h growth assays performed on the ancestral, control and adapted populations, (NS = not significant). The F-ratio is computed from the selection treatment MS/error MS; concentration MS/error MS and the interaction (selection x concentration) MS/error MS [29].