GC-MS-Based Metabolomics Study of Single- and Dual-Species Biofilms of Candida albicans and Klebsiella pneumoniae
Abstract
:1. Introduction
2. Results and Discussion
3. Materials and Methods
3.1. Strains Cultivation
3.2. Development and Quantification of Mono- and Polymicrobial Biofilms
3.3. Quantification of Viable Cells in the Biofilms
3.4. Metabolomic Analysis
3.4.1. Sample Preparation
3.4.2. GC-MS Analysis
3.4.3. Data Processing and Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Pairwise Comparisons | Univariate Statistical Test | Multivariate Analysis | R2X | R2Y | Q2Y |
---|---|---|---|---|---|
(C. albicans planktonic) vs. (C. albicans biofilm) | Student’s t-test | PLS-DA | 0.7287 | 0.9996 | 0.9876 |
(K. pneumoniae planktonic) vs. (K. pneumoniae biofilm) | Student’s t-test | PLS-DA | 0.6749 | 0.9983 | 0.9663 |
(C. albicans/K. pneumoniae biofilm 24 h) vs. (C. albicans biofilm 24 h) | Student’s t-test | PLS-DA | 0.7348 | 0.9994 | 0.9803 |
(C. albicans/K. pneumoniae biofilm 24 h) vs. (K. pneumoniae biofilm 24 h) | Student’s t-test | PLS-DA | 0.6368 | 0.9952 | 0.9261 |
(C. albicans/K. pneumoniae biofilm 24 h) vs. (C. albicans/K. pneumoniae biofilm 48 h) | Student’s t-test | PLS-DA | 0.5975 | 0.9988 | 0.9988 |
Dual Species vs. C. albicans | Dual Species vs. K. pneumoniae | |||||
---|---|---|---|---|---|---|
Name | VIP Score | t-Test p-Value | Fold Change | VIP Score | t-Test p-Value | Fold Change |
Acetamide, N,N-diethyl- (RI: 1045) | 0.864 | 3.10 × 10−2 | =2.24 | 0.264 | 6.70 × 10−2 | =1.13 |
Adenine, TMS (RI: 1890) | 1.320 | 2.86 × 10−9 | ↑ 25.63 | 1.561 | 3.12 × 10−9 | ↑ 19.85 |
Alanine, 2TMS (RI: 1124) | 0.623 | 1.76 × 10−1 | =1.15 | 1.076 | 2.16 × 10−2 | ↑ 1.24 |
Arabitol, 5TMS (RI: 1750) | 0.606 | 1.78 × 10−1 | =1.28 | 1.573 | 4.22 × 10−13 | ↑ 883.44 |
Asparagine, 3TMS (RI: 1687) | 1.125 | 2.56 × 10−3 | ↓ 1.24 | 1.490 | 1.26 × 10−7 | ↓ 1.73 |
Aspartic acid, 3TMS (RI: 1540) | 0.773 | 6.12 × 10−2 | =1.59 | 0.454 | 3.94 × 10−1 | =1.16 |
Beta-lactic acid, 2TMS (RI: 1156) | 0.359 | 4.38 × 10−1 | =1.17 | 1.278 | 2.32 × 10−3 | ↑ 1.76 |
Butanoic acid, 3TMS (RI: 1425) | 0.829 | 4.17 × 10−2 | =2.24 | 0.385 | 5.92 × 10−1 | =1.03 |
Citric acid, 4TMS (RI: 1844) | 0.659 | 1.28 × 10−1 | =1.46 | 0.706 | 1.73 × 10−1 | =1.58 |
Cyclo-(Phe-Pro) (RI: 2434) | 1.308 | 5.26 × 10−8 | ↓ 8.84 | 1.293 | 1.87 × 10−3 | ↓ 4.51 |
3-Deoxy-D-arabino-hexonic acid γ-lactone, 3TMS (RI: 1797) | – | 1.276 | 2.37 × 10−3 | ↓ 1.30 | ||
Glutamic acid, 3TMS (RI: 1638) | 1.062 | 3.35 × 10−3 | ↑ 1.33 | 0.778 | 1.28 × 10−1 | =1.12 |
Glyceric acid, 3TMS (RI: 1346) | 0.946 | 1.76 × 10−2 | =1.34 | 1.538 | 1.73 × 10−7 | ↓ 4.81 |
Glycerol, 3TMS (RI: 1290) | 1.002 | 7.48 × 10−3 | ↓ 1.33 | 1.524 | 1.17 × 10−6 | ↑ 644.01 |
Glycine, 2TMS (RI: 1136) | 1.274 | 3.46 × 10−6 | ↓ 1.83 | 0.223 | 9.37 × 10−1 | =1.01 |
Glycolic acid (2TMS) (RI: 1100) | 1.040 | 4.75 × 10−3 | ↓ 2.15 | – | ||
Isoleucine, 2TMS (RI: 1307) | 0.421 | 3.56 × 10−1 | =1.60 | 0.694 | 1.82 × 10−1 | =1.60 |
Lactic acid, 2TMS (RI: 1083) | 0.437 | 3.42 × 10−1 | =10.26 | 0.971 | 4.57 × 10−2 | =7.38 |
Leucine, 2TMS (RI: 1286) | 0.422 | 3.50 × 10−1 | =1.32 | 1.189 | 7.00 × 10−3 | ↑ 523.57 |
Lysine, 3TMS (RI: 1722) | 0.576 | 1.98 × 10−1 | =1.23 | 1.212 | 6.35 × 10−3 | ↑ 1.41 |
Methionine, 2TMS (RI: 1536) | 1.101 | 1.70 × 10−3 | ↓ 1.21 | 0.374 | 7.28 × 10−1 | =1.02 |
Nicotinic acid, TMS (RI: 1304) | 1.304 | 1.22 × 10−7 | ↓ 2.51 | 1.231 | 4.39 × 10−3 | ↓ 1.36 |
Ornithine, 3TMS (RI: 1632) | 1.161 | 4.61 × 10−4 | ↑ 4.34 | 0.583 | 2.86 × 10−1 | =1.20 |
Phenylalanine, 2TMS (RI: 1647) | 1.224 | 6.09 × 10−5 | ↑ 1.26 | 0.884 | 8.34 × 10−2 | =1.07 |
Phosphate, 3TMS (RI: 1297) | 0.564 | 2.12 × 10−1 | =1.48 | 0.242 | 7.02 × 10−1 | =1.11 |
Piperidine-2-carboxylic acid, 2TMS (RI: 1624) | 1.239 | 3.40 × 10−5 | ↓ 2.69 | 1.299 | 1.78 × 10−3 | ↓ 1.38 |
Proline, 2TMS (RI: 1314) | 1.282 | 1.88 × 10−6 | ↓ 11.44 | 1.127 | 1.34 × 10−2 | ↓ 3.60 |
Propanoic acid, 2methyl-2,3bishidroxy, 3TMS (RI: 1336) | 1.210 | 1.08 × 10−4 | ↓ 5.16 | – | ||
Pyroglutamic acid, 2TMS (RI: 1546) | 0.444 | 3.28 × 10−1 | =1.23 | 0.669 | 2.06 × 10−1 | =1.08 |
Pyruvic acid, 2TMS (RI: 1108) | 0.633 | 1.67 × 10−1 | =2.91 | 0.367 | 5.47 × 10−1 | =1.19 |
Serine, 3TMS (RI: 1375) | 1.320 | 1.49 × 10−9 | ↓ 307.99 | – | ||
Shikimic acid, 4TMS (RI: 1951) | – | 1.476 | 1.71 × 10−5 | ↓ 2.98 | ||
Succinic acid, 2TMS (RI: 1322) | 1.205 | 1.34 × 10−4 | ↓ 1.77 | 0.141 | 1 | =1.00 |
Threonine, 3TMS (RI: 1400) | 0.730 | 9.76 × 10−2 | =1.14 | 1.540 | 1.58 × 10−7 | ↓ 1.71 |
Trehalose, 8TMS (RI: 2781) | 1.127 | 1.06 × 10−3 | ↓ 10.10 | – | ||
Tryptophan, 3TMS (RI: 2244) | 1.138 | 4.53 × 10−4 | ↑ 1.90 | 1.119 | 1.29 × 10−2 | ↑ 1.28 |
Tyrosine, 3TMS (RI: 1962) | 1.293 | 3.24 × 10−6 | ↑ 1.81 | 1.323 | 3.28 × 10−4 | ↑ 1.21 |
Uracil, 2TMS (RI: 1351) | 0.968 | 1.20 × 10−2 | =1.67 | 0.901 | 1.05 × 10−1 | =1.38 |
Valine, 2TMS (RI: 1230) | 1.043 | 4.41 × 10−3 | ↓ 1.16 | 0.512 | 3.37 × 10−1 | =1.05 |
Vitamin B6, 3TMS (RI: 1924) | 0.660 | 7.28 × 10−1 | =1.36 | 0.564 | 1.76 × 10−1 | =1.09 |
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Galdiero, E.; Salvatore, M.M.; Maione, A.; de Alteriis, E.; Andolfi, A.; Salvatore, F.; Guida, M. GC-MS-Based Metabolomics Study of Single- and Dual-Species Biofilms of Candida albicans and Klebsiella pneumoniae. Int. J. Mol. Sci. 2021, 22, 3496. https://doi.org/10.3390/ijms22073496
Galdiero E, Salvatore MM, Maione A, de Alteriis E, Andolfi A, Salvatore F, Guida M. GC-MS-Based Metabolomics Study of Single- and Dual-Species Biofilms of Candida albicans and Klebsiella pneumoniae. International Journal of Molecular Sciences. 2021; 22(7):3496. https://doi.org/10.3390/ijms22073496
Chicago/Turabian StyleGaldiero, Emilia, Maria Michela Salvatore, Angela Maione, Elisabetta de Alteriis, Anna Andolfi, Francesco Salvatore, and Marco Guida. 2021. "GC-MS-Based Metabolomics Study of Single- and Dual-Species Biofilms of Candida albicans and Klebsiella pneumoniae" International Journal of Molecular Sciences 22, no. 7: 3496. https://doi.org/10.3390/ijms22073496