The effect of seminal pathogens on standard semen parameters, sperm kinematics and seminal inflammatory markers.

This study evaluated the effects of urogenital pathogens on standard semen parameters, sperm kinematics and host inflammatory response in a cohort of asymptomatic subfertile men. There were six groups based on the results of bacterial culture, including Ureaplasma urealyticum (U. Urealyticum) (n = 27), mixed comprising two or more pathogenic species (n = 28), Gardnerella Vaginalis (G. Vaginalis) (n = 15), gram-positive cocci and bacilli (g+cocci/bacilli) (n = 15), gram-negative bacilli (g-bacilli) (n = 10) and Chlamydia trachomatis (C. trachomatis) (n = 2). One control group (n = 20) and one leukocytospermic group (n = 10) were also included. Sperm quality parameters, seminal leukocytes and interleukin (IL)-6 of all groups, apart from C. trachomatis, were compared to the control group. Standard semen parameters were significantly worse in all groups except for that with g-bacilli. Progressive motility, total motility and normal sperm morphology demonstrated the most significant differences, when U. Urealyticum, leukocytospermia and mixed pathogens were detected in semen. Among sperm kinematics, the concentration of progressive motile sperm cells (CPMS), the percentage of progressive motile sperm cells (PPMS) and straightness (STR) were manifested significant declines in the presence of seminal pathogens. CPMS was affected in all groups except for G. vaginalis. Moreover, the presence of g+cocci/bacilli and g-bacilli were associated with increased seminal IL-6. Seminal leukocytes were elevated significantly only when g-bacilli were cultured in semen. We conclude that seminal pathogens can negatively affect sperm quality. The most negative effect is related to U. Urealyticum. Moreover, g+cocci/bacilli and g-bacilli can initiate an inflammatory response.

Standard Semen Parameters vs. Sperm Kinematics to Predict Sperm DNA Damage.

PURPOSE: The aims of this study were to associate sperm kinematics and standard semen parameters with sperm DNA damage and to evaluate whether the addition of sperm kinematics improve the multivariable prediction of sperm DNA fragmentation compared to standard semen parameters alone. MATERIALS AND METHODS: We evaluated sperm kinematics, standard semen parameters, and DNA fragmentation index (DFI) in 122 men. Univariate and multivariate logistic regression models were fitted to evaluate the association of sperm kinematics and standard semen parameters with pathologically damaged sperm DNA (DFI≥26%), and receiver operating characteristics (ROC) curves were calculated for these models. RESULTS: On univariate analyses, average velocity, curvilinear velocity, straight-line velocity, straightness (STR), beat-cross frequency (BCF), and the percentage of progressive motile sperm cells (PPMS) were significantly associated with pathologically damaged sperm DNA. Likewise, among standard semen parameters, sperm concentration, progressive motility, normal morphology, and vitality were found to be linked with sperm DNA damage. On the multivariate analysis, vitality was the strongest predictor of pathologically damaged sperm DNA with an area under the ROC curve (AUROC) of 88.3%. Adding STR, BCF, and PPMS to vitality increased the AUROC to the significant extent of 91.5%. CONCLUSIONS: Sperm vitality is the most accurate routine-based laboratory test for the prediction of pathologically damaged sperm DNA, but the addition of sperm kinematics increases its accuracy. Both standard semen parameters and sperm kinematics are complementary in predicting pathologically damaged sperm DNA, and might serve as a new tool to screen for fertile men.

The 1999 and 2010 WHO reference values for human semen analysis to predict sperm DNA damage: A comparative study.

Standard semen parameters are often used to predict male fertility, but whether the 2010 World Health Organization (WHO 2010) thresholds are better predictors than the 1999 thresholds has not been investigated. In this study, we addressed this issue using sperm DNA fragmentation (SDF) as a marker of male fertility in 134 subfertile male individuals. To compare the predictive value of the 1999 thresholds with the 2010 cutoffs, the Youden indices (YIs) of all possible thresholds were calculated using receiver operating characteristic (ROC) curves and compared to each other and to the respective YIs of optimal thresholds. We found that the area under the ROC curves of progressive motility and vitality was the highest among standard semen parameters, and that the YI of both parameters from the 2010 manual was comparable to the respective optimal YIs. In contrast, the threshold of sperm concentration and total sperm number from both WHO recommendations demonstrated low YIs, with substantial differences to the respective optimal YIs. The YIs of normal morphology cutoffs from both WHO manuals were slightly different from each other and from the respective optimal YIs. In conclusion, the 2010 thresholds for progressive motility and vitality are superior to the 1999 thresholds in predicting SDF, whereas the cutoff value of sperm concentration, total sperm number and normal morphology may need further revisions to increase their accuracy.

The association of seminal leucocytes, interleukin-6 and interleukin-8 with sperm DNA fragmentation: A prospective study.

The effect of seminal leucocytes on sperm DNA integrity has been discussed controversially in literatures. Moreover, the studies investigating the in vivo effect of pro-inflammatory cytokines interleukin-6 and interleukin-8 on sperm DNA fragmentation are scarce and inconsistent. The association of standard sperm parameters with sperm DNA fragmentation is also a matter of ongoing discussion. Hence, the aims of this study were, first, to evaluate the effect of seminal leucocytes, interleukin-6 and interleukin-8 on sperm DNA integrity and, second, to examine whether standard semen parameters are associated with sperm DNA fragmentation. Seminal leucocytes, interleukin-6, interleukin-8 and standard semen parameters, including total sperm number, sperm concentration, progressive motility, nonprogressive motility, immotility and normal morphology, were determined in 134 consecutive men. The concentrations of seminal leucocytes, interleukin-6 and interleukin-8, did not correlate with sperm DNA fragmentation. In contrast, total sperm number, sperm concentration, progressive motility, nonprogressive motility and normal morphology exhibited significant inverse correlations with sperm DNA fragmentation. Immotile spermatozoa were directly correlated with sperm DNA fragmentation. In conclusion, seminal leucocytes, interleukin-6 and interleukin-8, are not associated with sperm DNA fragmentation. Poor standard semen parameters are significantly related to the high levels of sperm DNA fragmentation.

Evaluation of Leukocyte Threshold Values in Semen to Detect Inflammation Involving Seminal Interleukin-6 and Interleukin-8.

OBJECTIVE: To evaluate leukocyte threshold values in semen to detect inflammation involving seminal interleukin (IL)-6 and IL-8. MATERIALS AND METHODS: The levels of leukocytes, IL-6, and IL-8 in semen were determined. The 75th and 90th percentiles of seminal IL-6 and IL-8 were considered as "high" and "very high" concentrations, respectively. Inflammatory semen was defined based on high levels of IL-6 (≥86.75 pg/mL) or IL-8 (≥4460 pg/mL). Very high levels of IL-6 (≥228 pg/mL) or IL-8 (≥12,480 pg/mL) were used to define acute seminal inflammation. On the basis of high and very high levels of IL-6 or IL-8, receiver operating characteristic curves were generated to evaluate leukocyte threshold values. RESULTS: Leukocytes at a cutoff level of 1 × 10(6)/mL had 51% sensitivity and 95% specificity to detect high levels of IL-6, whereas on the basis of very high levels of IL-6, the same cutoff level revealed 82% sensitivity and 90% specificity. Similarly, leukocytospermia demonstrated low sensitivity (56%) to detect high levels of IL-8 but acceptable sensitivity (94%) and specificity (92%) to predict very high levels of IL-8. The cutoff level of 0.315 × 10(6) leukocytes/mL had optimal sensitivity and specificity for predicting high levels of inflammatory cytokines. CONCLUSION: Leukocytospermia demonstrated poor sensitivity to detect seminal inflammation, as defined by high levels of inflammatory cytokines. The optimal threshold value to detect inflammation was found to be 0.315 × 10(6) leukocytes/mL. On the basis of very high levels of IL-6 or IL-8, leukocytospermia is a sensitive and specific marker to predict acute seminal inflammation.

Influence of pathogens and moderate leukocytes on seminal interleukin (IL)-6, IL-8, and sperm parameters.

PURPOSE: To evaluate the role of pathogens and moderate leukocytes on seminal interleukin (IL)-6, IL-8, and sperm parameters in men undergoing infertility investigation. METHODS: Semen samples from men (n = 171) were divided into three groups on the basis of leukocyte count: no leukocytes (L-; ≤ 0.1 × 10(6)/ml Mio/ml), moderate leukocytes (L ±; >0.1 × 10(6)/ml and <1 × 10(6)/ml), and high leukocytes (=leukocytospermia) (L+; ≥ 1 × 10(6)/ml). Each group was further classified into two subgroups, according to the presence (B+) or absence (B-) of pathogens. IL-6, IL-8, and sperm characteristics were analyzed in each subgroup. A correlation test was performed to show the association between inflammatory parameters and sperm characteristics. RESULTS: No significant differences in leukocyte count, cytokine levels, and sperm characteristics were apparent in subgroups with and without pathogens. Grade b motility was significantly lower in subgroup IIa (L ±,B-) than in subgroup Ia (L-,B-)(p < 0.05). More significant limitations in sperm motility (lower rapid progressive motility and increased percentage of immotile sperm) were observed in subgroup IIIa (L+,B-) compared with subgroup Ia (p < 0.05). Moderate and high leukocytes increased significantly cytokine levels (p < 0.001). CONCLUSIONS: Moderate leukocyte counts could be an indicator of male genital tract inflammation. Seminal pathogens have no influence on cytokine levels and sperm parameters.

New method for differentiating chronic prostatitis/chronic pelvic pain syndrome IIIA from IIIB involving seminal macrophages and monocytes.

OBJECTIVES: To evaluate a new method for differentiating inflammatory from noninflammatory prostatitis using the simple and rapid quantification of seminal macrophages and monocytes. METHODS: Patients affected with chronic pelvic pain syndrome (CPPS) were classified as having the IIIA (n = 11) and IIIB (n = 30) subtypes according to the peroxidase positive leukocyte concentration in semen; 18 healthy individuals served as controls. Seminal inflammatory markers, including polymorphonuclear elastase, interleukin (IL)-6 and IL-8, and numbers of macrophages/monocytes (MMs) per 50 fields of 1000 × magnification (high-power field [hpf]), were determined for all patients. RESULTS: The numbers of MMs/50 hpf correlated significantly with the peroxidase positive leukocyte counts and IL-8, IL-6, and polymorphonuclear elastase levels (all P < .001). Data from the analysis of receiver operating characteristic curves (area under the curve 0.912 ± 0.073; P < .001) showed a sensitivity of 90.9% and specificity of 86.7% at a cutoff value of 5 MMs/50 hpf. The positive and negative predictive value was 71.4% and 96.3%, respectively. The median concentrations of IL-6, IL-8, and elastase in the patients with CPPS with ≥ 5 MMs/50 hpf differed significantly (P ≤ .002) from those in the patients with <5 MMs/50 hpf. CONCLUSIONS: The results of our study have shown that the quantification of seminal macrophages and monocytes is a simple, rapid, and reproducible technique by which to differentiate chronic prostatitis/CPPS IIIA from IIIB.