TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick-end labeling (TUNEL) assay of spermatozoa.

<p>Histograms show: (<b>A</b>) negative control with 2.35% TUNEL positive cells. (<b>B</b>) Positive control (spermatozoa treated with DNaseI) with 90.5% TUNEL positive cells. (<b>C</b>) Semen sample of one male partner of infertile couples positive for <i>C. trachomatis</i> qPCR with 20.5% TUNEL positive cells. B: window adjusted to detect the percentage of TUNEL positive cells.</p

Primers and probes used for detection and quantification of C. trachnomatis, N. gonorrhoeae. U. urealyticum, M. genitalium, U. parvum and M. hominis by qPCR.

<p>Primers and probes used for detection and quantification of C. trachnomatis, N. gonorrhoeae. U. urealyticum, M. genitalium, U. parvum and M. hominis by qPCR.</p

Seminological variables of semen of <i>C. trachomatis</i> positive patients compared to uninfected patients.

<p>Note: Values are means (± Standard Error).</p>#<p>Unless indicated, variables were tested T-Test.</p

Flow cytometric of sperm viability using 7-amino-actinomycin-D Dye.

<p>Histograms show: (<b>A</b>) Negative control with 10% 7-AAD positive cells. (<b>B</b>) Positive control with 98.5% 7-AAD positive cells. (<b>C</b>) Semen sample of one male partner of infertile couples positive for <i>C. trachomatis</i> qPCR with 56.5% 7-AAD negative cells and 43.5% 7-AAD positive cells. B: window adjusted to detect the percentage of cells with 7-AAD positive.</p

Distributions of percentages of different apoptotic markers among patients positive for <i>C. trachomatis</i> qPCR compared to uninfected patients.

<p>(<b>A</b>) Mean percentage of Sperm Vitality, evaluated with 7-amino-actinomycin-D Dye (7-AAD). (<b>B</b>) Mean percentage of Sperm mitochondrial membrane potential (ΔΨm), evaluated with JC-1. (<b>C</b>) Mean percentage of Caspase 3 activation, evaluated with fluorescein-labeled inhibitor of caspases (FLICA). (<b>D</b>) Mean percentage of Sperm DNA fragmentation, evaluated with (TUNEL). <b>Uninfected</b>: Sperm of uninfected patients (negative for all PCRs performed and for spermioculture analysis). <b>CT⁺</b>: sperm of patients positive for <i>C. trachomatis</i> qPCR. * Indicates significant differences compared with uninfected semen (<i>P</i><0.05). ** Indicates significant differences compared with uninfected semen (<i>P</i><0.001).</p

Flow cytometric of changes in the mitochondrial membrane potential (ΔΨm).

<p>Histograms show: (<b>A</b>) Negative control with 95.8% low ΔΨm cells. (<b>B</b>) Positive control with 9.6% low ΔΨm cells. (<b>C</b>) Semen sample of one male partner of infertile couples positive for <i>C. trachomatis</i> qPCR with 32.5% low ΔΨm cells and 67.5% with high ΔΨm cells. C: window adjusted to detect the percentage of cells with low ΔΨm.</p

Frequency of urogenital bacteria detected by qPCR and spermiocultures analysis in semen samples of 85 infertile male patients.

<p>Frequency of urogenital bacteria detected by qPCR and spermiocultures analysis in semen samples of 85 infertile male patients.</p