Table 3 Key metabolites in both skin and liver tissues discriminating between experimental groups (including sampling periods post exposure and populations), demonstrating overlap in significant results between univariate (ANOVA, t-test pattern matching and SAM) and multivariate analyses (PLS-DA, top 30 VIP scores ranked on component 1).

From: Chytridiomycosis causes catastrophic organism-wide metabolic dysregulation including profound failure of cellular energy pathways

Group

Tissue

Metabolite identification

(TMS groups, RT, RI)

ANOVA FDR

Tukey’s HSD

Pattern matching

FDR

Correlation

SAM FDR

PLS-DA VIP

comp. 1

Time-series

Skin

a-Ketoglutaric acid, x TMS, 23.95, 1578

1.05E-05

4-0; 4-1; 4-2; 4-3

0.005854

−0.50794

0

2.2876

Serotonin, x TMS, 39.11, 2470

0.002533

4-0; 4-1; 4-2; 4-3

  

0

1.6147

Putrescine, x TMS, 22.45, 1506_putative

0.036876

3-0; 4-0

0.005854

0.50557

0.00909

2.0726

Adenine, 2 TMS, 29.74, 1869

0.036337

3-0; 4-0

0.005854

0.50493

0.00909

2.0262

5-Hydroxyindole-3-acetic acid, 3 TMS, 35.49, 2212

0.002533

4-0; 4-1; 4-2; 4-3

  

0.000346

 

L-Threonine, 3 TMS, 19.59, 1387

0.02635

3-0; 4-0

0.012166

−0.472

0.003239

1.99

L-Isoleucine, 2 TMS, 17.32, 1295

0.004158

4-0; 4-1; 4-2; 4-3

  

0.004607

 

DL-Tartaric acid 4TMS-like

0.021251

4-0; 4-1; 4-2

0.039136

−0.41978

0.00909

 

Glutamic acid, 3 TMS, 24.79, 1623

0.004834

4-0; 4-1; 4-2; 4-3

0.039136

−0.40884

0.003179

 

Adenosine, 4 TMS, 41.49, 2642

  

0.039136

0.41253

0.042185

 

Urea, 2 TMS, 16.14, 1249

0.02635

1-0; 3-1

  

0.000829

 

Serine, 2 TMS, 16.43, 1260

0.029108

3-0; 3-2

  

0.003179

1.719

DL-Ornithine, 3 TMS, 24.71, 1623

0.012299

2-1; 3-2; 4-2

  

0.003293

 

L-Pyroglutamic acid, 2 TMS, 22.76, 1520_saturated

0.032369

4-0; 4-2

0.039136

−0.39293

0.019789

 

L-Lysine, 4 TMS, 30.54, 1915

0.036876

2-1; 3-1; 4-1

  

0.006945

1.7061

Fumaric acid, 2 TMS, 18.29, 1357

0.02968

4-0; 4-3

  

0.027929

 

Aspartic acid, 2 TMS, 20.54, 1428

    

0.018362

1.651

Cellobiose, x TMS, 42.19, 2962

  

0.040882

0.38103

0.039192

1.496

g-Aminobutyric acid, 3 TMS, 22.9, 1526

0.0409

4-0; 4-1; 4-3

  

0.00909

 

Myo-Inositol, 6 TMS, 33.38, 2081

  

0.042804

−0.37252

0.032292

 

Liver

L-Isoleucine, 2 TMS, 17.32, 1295

3.51E-11

4-0; 2-1; 4-1; 3-2; 4-2; 4-3

4.15E-05

0.57373

0.003774

1.5512

L-Glutamic acid, 2 TMS, 22.7, 1519*

2.86E-10

4-0; 4-1; 4-2; 4-3

2.50E-06

−0.6347

0.011043

1.5014

5-Hydroxyindole-3-acetic acid, 3 TMS, 35.49, 2212

0.000582

4-0; 4-1; 4-2; 4-3

0.001556

0.45911

0.003029

2.5384

D–Galactose, 5 TMS, MEOX, 29.94, 1880

0.025509

4-0; 4-1

0.011265

−0.38718

0.003029

2.6805

L-Pyroglutamic acid, 2 TMS, 22.76, 1520_saturated

2.86E-10

4-0; 4-1; 4-2; 4-3

2.50E-06

−0.63459

0.011043

1.501

Citric acid, 4 TMS, 28.69, 1817

9.67E-07

4-0; 4-1; 4-2; 4-3

0.000752

−0.4873

0.003029

1.966

Putrescine, x TMS, 22.45, 1506_putative

5.64E-07

4-0; 4-1; 4-2; 4-3

6.71E-06

0.61232

0.003029

2.5008

a-Ketoglutaric acid, x TMS, 23.95, 1578

8.29E-05

4-0; 4-1; 4-2; 4-3

0.002517

−0.44337

0.003029

1.9493

L-Leucine, 2 TMS, 16.74, 1274

1.85E-08

4-0; 4-1; 3-2; 4-2; 4-3

4.18E-05

0.57127

0.011264

 

Creatine. x TMS, 23.39, 1551

6.32E-05

4-0; 4-1; 4-2; 4-3

0.000752

0.48784

0.003029

1.9563

D-Fructose-6-phosphate 6TMS, MEOX, 36.72, 2300-put

5.97E-07

4-0; 4-1; 4-2; 4-3

0.002165

−0.44874

0.003029

1.8381

g-Aminobutyric acid, 3 TMS, 22.9, 1526

4.26E-05

4-0; 4-1; 3-2; 4-2

0.000315

−0.51611

0.004805

1.7973

D-Glucose-6-phosphate, 6 TMS, MEOX, 37.23, 2332

3.10E-06

4-0; 4-1; 4-2; 4-3

0.006038

−0.41173

0.003029

 

Fumaric acid, 2 TMS, 18.29, 1357

1.42E-06

4-0; 4-1; 4-2; 4-3

0.000934

−0.47769

0.005732

 

Serine, 3 TMS, 18.98, 1363

0.003442

3-0; 4-0; 4-1; 4-2

0.000661

0.49314

0.038474

 

L-Lysine, x TMS, 29.32, 1852

6.32E-05

4-0; 4-1; 4-2; 4-3

0.003008

0.4358

0.00402

1.5988

Urea, 2 TMS, 16.14, 1249

    

0.011043

1.5188

Glycine, 3 TMS, 17.63, 1308

0.048096

4-0

0.003008

0.43531

  

Sucrose, 8 TMS, 41.32, 2630.1

0.005833

1-0; 2-0; 3-1; 3-2

  

0.008361

 

Adenine, 2 TMS, 29.74, 1869

0.016867

4-0; 4-1; 4-2; 4-3

  

0.011043

 

Site

Skin

Pantothenic acid, O,O,O-TMS-putative

0.012106

G-E; K-E

0.010965

−0.51381

0.002488

2.1179

Myo-Inositol, 6 TMS, 33.38, 2081

0.017097

K-E; K-G

  

0.0199

 

L-Isoleucine, 2 TMS, 17.32, 1295

0.026777

K-E

0.010965

0.49005

  

L-Leucine, 2 TMS, 16.74, 1274

0.026777

K-E; K-G

0.010965

0.48099

0.039608

1.5988

Creatine. x TMS, 23.39, 1551

0.046951

K-E; K-G

0.016922

0.45692

0.036566

1.8447

Aspartic acid, 2 TMS, 20.54, 1428

     

1.9701

L-Threonine, 2 TMS, 17.37, 1298

     

1.8778

D–Ribose, 4 TMS, MEOX, 25.89, 1678

  

0.033096

0.42058

  

saturated_Guanine manual

     

1.7444

Serotonin, x TMS, 39.11, 2470*

     

1.7352

Liver

D–Turanose, 7 TMS, 42.29, 2702

     

2.7638

Urea, 2 TMS, 16.14, 1249

     

2.507

Thymine, 2 TMS, 19.94, 1403

     

2.4842

L-Lysine, 3 TMS, 26.54, 1712

     

2.0644

Myo-Inositol, 6 TMS, 33.38, 2081.1

     

1.8873

D-Fructose-6-phosphate 6TMS, MEOX, 36.72, 2300-putative

     

1.7514

Sucrose, 8 TMS, 41.32, 2630.1

     

1.734

a-Ketoglutaric acid, x TMS, 23.95, 1578

     

1.6361

beta-Alanine, 3TMS, 20.58, 1429

     

1.6332

Cellobiose, x TMS, 42.19, 2962

     

1.4796

  1. Comparisons found to be significant with post-hoc Tukey’s HSD tests are listed, as are the respective correlations for the pattern matching approach. Sham-exposed negative control group frogs were designated Group 0. Exposed/infected frogs sampled at various times post-exposure were designated Groups 1, 2, 3 and 4, corresponding to being sampled at 4, 8, 14 and 28–30 days post exposure, respectively. Source populations (sites) are as follows: Eucumbene, Grey Mare, Kiandra are represented by E, G and K respectively. Where metabolites were not found to be significantly different with a particular test, values have been omitted.
  2. *Quantitation re-calculated from a lesser abundant ion (or isotope) within mass spectrum for accurate measurement of metabolites nearing the upper dynamic concentration of the MS.
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