Relative change in free amino acid concentration during growth of <i>P</i>. <i>freudenreichii</i> subsp. <i>shermani</i> CIRM BIA 1 in CdM (A, C and E) and EJ (B, D and F).

<p>(A and B) Alliphatic amino acids; (C and D) basic, acidic + amide amino acids; (E and F) Aromatic, hydroxyl and sulfur-containing amino acids. The arrows indicate the main growth phases corresponding to each incubation medium: exponential (Exp), early stationary (E.Stat) and late stationary phase (L.Stat).</p

Two-dimensional analysis of protein expression during growth of <i>P</i>. <i>freudenreichii</i> subsp. <i>shermani</i> CIRM BIA 1 in EJ and in CdM.

<p>(A) Neosynthesized proteins were radiolabeled with ³⁵S amino acids in both media during indicated duration: lag phase (early), the beginning of the exponential phase (early exponential), the exponential and early stationary phases. (B) Cellular proteins accumulated during growth were Coomassie blue stained.</p

<i>P</i>. <i>freudenreichii</i> subsp. <i>shermani</i> CIRM BIA 1 tolerance to digestive stresses.

<p>Propionibacteria were harvested at different stages of growth in CdM black square or in EJ white square, prior to acid (A, pH 2) or bile salts (B, 1 g L^-1) challenges. Surviving bacteria were then counted by CFU enumeration. Means with different lower case superscript letters (a-b) differ significantly (<i>P</i> < 0.05) in italic groups for CdM and in normal case groups for EJ.</p

Growth and survival of <i>P</i>. <i>freudenreichii</i> subsp. <i>shermani</i> CIRM BIA 1 in EJ and CdM

<p>(A) growth curve and (B) Live/dead staining of the bacterial cells in the exponential (Exp), early stationary (E.Stat) and late stationary phase (L.Stat) of growth in CdM (<b>black dot</b>) and in EJ (<b>white dot</b>).</p

Proteins differentially expressed after growth of <i>P</i>. <i>freudenreichii</i> subsp. <i>shermani</i> CIRM BIA 1 in EJ and in CdM and identified by on line coupling NanoLC-ESI-Q-TOF tandem mass spectrometry.

<p>*: Fold changes correspond to the protein overexpressed with a minimum fold change of 1.2 in the CdM or EJ determined by image analysis</p><p>**: e-value is the number of times a given peptide score will be achieved by incorrect matches from a database search. Protein identifications were automatically validated when they showed at least two unique peptides with an e-value below 0.05 corresponding to log(e-value) < -1.30</p><p>Proteins differentially expressed after growth of <i>P</i>. <i>freudenreichii</i> subsp. <i>shermani</i> CIRM BIA 1 in EJ and in CdM and identified by on line coupling NanoLC-ESI-Q-TOF tandem mass spectrometry.</p

Cell viability with various centrifugation rates (400, 1500, 3000, 5000×<i>g</i> during 10 min at 4°C) comparing to the reference milk cell suspension without supplementary centrifugation (Ref).

<p>(A) Boxplot and whiskers of flow cytometry results with milk somatic cell suspensions incubated with specific antibodies and vioblue live/dead kit (n = 4); (B) Cell viability of each type cell for 400×<i>g</i> (white bars), 1500×<i>g</i> (hatched bars), 3000×<i>g</i> (grey bars), 5000×<i>g</i> (dark bars) centrifugations by flow cytometry; (C) Mean proportion of macrophages (dark sectors), PMNs (grey sectors) and lymphocytes (white sectors) in cell samples under various centrifugation with different gravitational velocities. Means with different superscripts (a-d) differ significantly (<i>P</i><0.05).</p

Flow cytometry identification of differential somatic cell count and simultaneous their quantification of all cells and each cell type.

<p>(A)(B)(C) fresh blood cell suspension, (D)(E)(F) milk cell suspension after 80°C × 30 min and (G)(H)(I)milk cell suspension conserved at 4°C in PBS pH 7.4. The cell subpopulations of each sample were shown in APC/FITC dotplot (A)(D)(G), the viable and non-viable population of all cells and each subpopulation were shown in histogram-Vioblue scatter (B)(E)(H) and (C)(F)(I), respectively.</p

Summary of the experimental design of the somatic cell preparation and the various treatments applied, i.e. storage in milk or PBS for 72 h, variation of the centrifugation rates and heat treatment.

<p>Summary of the experimental design of the somatic cell preparation and the various treatments applied, i.e. storage in milk or PBS for 72 h, variation of the centrifugation rates and heat treatment.</p

Flow cytometry scatter pattern for the identification of differential somatic cells in blood-milk mix cell suspension (1/1, v/v) and corresponding isotype control sample, respectively.

<p>(A) FSC/SSC dotplot of cell suspension. All somatic cells (in yellow) in cell suspension (B) were identified by CD45/PerCp+. The subpopulation of cell suspension in APC/FITC dotplot (C), macrophages (red), PMNs (blue) and lymphocytes (green) are identified by CD14/APC+ gate in APC/SSC plot (D), CH138A/FITC+ gate in FITC/SSC plot (E), and FSC/SSC size/granularity gate (F), respectively.</p