Opportunistic strains showed altered CN of specific genes.

<p>A) Genes that showed altered CNV in the three opportunistic strains were clustered and the heat map obtained (Mev software) is represented. B) Evaluation of growth with different concentrations of copper was performed in microtiter plates in YPD media. Maximal growth for each Cu concentration was relativized to YPD without Cu for each strain. Average and standard deviation of triplicates is represented C) Gene copy number was determined by qPCR. Data was normalized with haploid strain S288C. Average and standard deviation of triplicates is represented. D) Heat map detail of genes that belong to de novo purine biosynthetic pathway (Mev software).</p

Comparative genomic hybrization of opportunistic strains.

<p>D14, 60 and 102 were compared to laboratory strain W303 hybridizing DNA to S288C cDNA microarray. A) Log2 of normalized and filtered data of the four strains is represented according its chromosomal disposition in stack bar graph. Specific genes are depicted with an arrow. B) Absolute signal intensity of genes is represented against distance to telomere. Discontinuous line represents sliding window average (10x) of values in 1 Kbp. C) Number of genes in increased or decreased CNV represented for each strain. D) Venn diagram showing the number of specific or common genes for strains D14, 103 and 60. E) Local zoom of general graph presented in A) for genes <i>IMD3</i>, <i>IMD2</i> and <i>CUP1</i> chromosomal regions for opportunistic strains.</p

Growth at different temperatures of commercial and control <i>S. cerevisiae</i> strains.

<p>Ten-fold serial dilutions of the indicated strains were dropped on YPD plates and incubated for 24 h at 30, 37, 39 and 42°C. +++: growth in all the dilutions; ++: growth in the first two dilutions; +: growth in the first dilution; ±: growth in the first drop without dilution; –: no growth.</p

<i>S. cerevisiae</i> strains used as control and assays for which they were used.

(a)<p>(named D5 in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0098094#pone-0098094-t001" target="_blank">Table 1</a>);</p>(b)<p>Molecular characterization of these strains have been described previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0098094#pone.0098094-deLlanos5" target="_blank">[44]</a>;</p>(c)<p>Growth at different temperatures,</p>(d)<p>protease and phospholipase secretion, Ph (pseudohyphal) and invasive growth;</p>(e)<p>MAPK activation;</p>(f)<p>Adherence to plastics;</p>(g)<p><i>In vivo</i> study by intravenous inoculation of strains in BALB/c mice;</p>(h)<p>(<i>MAT</i><b>a</b><i>; ura3-52; trp1D2; leu2-3_112; his3-11; ade2-1; can1-100</i>) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0098094#pone.0098094-Wheeler1" target="_blank">[64]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0098094#pone.0098094-Thomas1" target="_blank">[76]</a>;</p>(i)<p>(<i>MAT</i><b>a</b><i>his3</i>Δ<i>1 leu2</i>Δ<i>0 met15</i>Δ<i>0 ura3</i>Δ<i>0)</i> (EUROSCARF); n.r.: Non reported.</p

Virulence-associated phenotypes observed in strains isolated from commercial products.

<p>Phenotype traits shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0098094#pone-0098094-g002" target="_blank">Figures (2</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0098094#pone-0098094-g004" target="_blank">4</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0098094#pone-0098094-g005" target="_blank">5</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0098094#pone-0098094-g006" target="_blank">6</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0098094#pone-0098094-g007" target="_blank">7</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0098094#pone-0098094-g008" target="_blank">8</a>) and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0098094#pone-0098094-t004" target="_blank">Table 4</a> were simplified as follows to calculate the Spearman’s Rho correlation coefficients between strains and apply the Jaccard proximity test (excluding joint absences for considerations): growth at 37, 39 and 42°C, +++, ++ and + were simplified to +, and ± and − were simplified to −; phospholipase activity (PLA), ++ and + were simplified to +; pseudohyphal (Ph) growth,++ and + were simplified to +; invasive growth, ± and − were simplified to −; switching, multiple colony phenotypes with frequencies between 10⁻²−10⁻⁴ were considered +; MAPK activation, high Slt2 and Kss1 phosphorylation was considered + and low or non phosphorylation were considered −; Plastics adherence (ADH), Petri plates adherence values >30% and microtiter plates absorbance values >1 were considered +, Petri plates adherence values ≤30% and microtiter plates absorbance values <1 were considered −. n.d. not determined.</p

Colony phenotype switching frequencies of commercial <i>Saccharomyces</i> strains.

<p>Colony phenotype descriptions: D2-A (5–7 mm, light pink, strong pink centre, white edge, creamy texture); D2-B (5–7 mm, light pink, strong pink centre, white edge, strong pink sector, creamy texture); D2-C (5–7 mm, strong pink, white edge, rough texture, striated); D2-D (3 mm, light pink, strong pink centre, creamy texture); D2-E (5–7 mm, light pink, strong pink centre, white edge, creamy texture, star shape); D2-F (5–7 mm, light pink, strong pink centre, white edge, white sector, creamy texture); D2-G (3 mm, strong pink, creamy texture, irregular); D2-H (4 mm, light pink, strong pink centre, white edge, strong pink sector, rough texture); D2-I (5–7 mm, light pink, strong pink centre, rough texture, striated); D4-A (5 mm, light pink, strong pink centre, creamy texture); D4-B (2 mm, light pink, strong pink centre, creamy texture, irregular); D4-C (1 mm, light pink, creamy texture, irregular); D4-D (3 mm, strong pink, creamy texture); D4-E (5 mm, light pink, strong pink centre, creamy texture, white sector); D4-F (5 mm, light pink, strong pink centre, creamy texture, irregular); D14-A (5 mm, light pink, pale pink centre, strong pink edge, creamy texture); D14-B (2–3 mm, pale pink, creamy texture); D14-C (2–3 mm, light pink, pale pink centre, creamy texture); D14-D (5 mm, light pink, pale pink centre, strong pink edge, creamy texture, white sector); D14-E (5 mm, light pink, pale pink centre, strong pink edge, creamy texture, strong pink sector); D14-F (5 mm, medium pink, pale pink centre, strong pink edge, creamy texture); D14-G (5 mm, light pink, pale pink centre, strong pink edge, creamy texture, striated); D14-H (2 mm, pale pink, dark pink edge, creamy texture); D23-A (4–5 mm, light pink, strong pink centre, creamy texture); D23-B (1–2 mm, strong pink, creamy texture); D23-C (4–5 mm, light pink, strong pink centre, creamy texture, irregular); D23-D (4 mm, strong pink, creamy texture); D23-E (7–8 mm, light pink, strong pink centre, creamy texture); D23-F (4–5 mm, light pink, strong pink centre, rough texture, striated); D23-G (5 mm, light pink, strong pink centre, strong pink sector, creamy texture); D23-H (1–2 mm, light pink, strong pink centre, creamy texture).</p

Distribution of DNA types of yeasts isolated from commercial products.

<p>Distribution of DNA types of yeasts isolated from commercial products.</p

Intestinal translocation and dissemination of isolates D14 and D23 based on the number of mice that showed yeast burdens in target organs.

(a)<p>Day of sacrifice after initiation of oral administration of the yeast.</p>(b)<p>MLNs: mesenteric lymph nodes.</p>(c)<p>Other organs: brain, kidneys, liver.</p

Generation time of commercial and control <i>S. cerevisiae</i> strains on YPD at 30°C and 37°C.

<p>Error bars correspond to standard deviations. ∧not determined. *<i>p</i><0.05 with regard to the avirulent strains (CECT 10.431 and W303), as assessed by Students t-test.</p

The critical role of genes related to dNTP pool increase in response to human blood phagocytes.

<p>A) Evaluation of yeast performance in response to 7 μg/ml of mycophenolic acid (MPA) or 0.004% of MMS was compared to control minimal media SC for opportunistic strains D14, 102 and 60 and for control lab strains S288C and W303. B) Evaluation of yeast survival after 90 min of <i>ex vivo</i> human blood incubation. After blood incubation, wild type lab strains BY4741 and W1588-4C and D14 opportunistic strain were plated in YPD or YPD with guanine (65 μM) and colony were counted. C) Also, dun1 and rnr1* mutants were plated in YPD after blood exposure. Triplicates average and standard deviation of normalized survival data (YPD + guanine respect YPD and mutants respect its isogenic parental wild type) is represented. In all cases (control respect guanine and mutants respect to wild type) differences were significantly different using <i>p</i>-value of 0.05.</p