Fluorescent in-situ hybridization (FISH) of decondensed sperm nuclei has been used directly to evaluate the enrichment efficiency of human sperm separation using Sephadex gel filtration and human serum albumin (HSA) gradients. Control and processed spermatozoa were fixed and their nuclei decondensed. In-situ hybridization was carried out with a Y-specific DNA probe (DYZ1). Sephadex filtration yielded 52.5% Y-chromosome-bearing spermatozoa, HSA separation resulted in 49.4% Y-chromosome-bearing spermatozoa and in the untreated control sample the percentage of Y spermatozoa was 49.3%. Statistical analysis revealed no significant differences between the selection methods employed and the controls, and no real enrichment for X- or Y-bearing spermatozoa was detected for any of the selection methods assayed. The usefulness of the protocols reported for selection of spermatozoa by sex chromosome in couples at risk for X-linked diseases is discussed.

A prospective study was carried out on 1005 male volunteers residing in the province of Barcelona. Participants were asked to complete a questionnaire concerning their health status and lifestyle, and provided a semen sample for analysis. Sperm concentration and percentages of motile, and normal forms were evaluated. The effect of smoking, alcohol consumption, stress and previous disease on semen parameters was also evaluated. Normal parameters were found in 22% of volunteers and sperm counts and/or percentage motility below normal threshold values were found in 78%. Asthenozoospermia detected in 62% and oligozoospermia in 17% of volunteers. No statistically significant association was found between semen quality and age, with the exception of a decrease in semen volume (P = 0.04) and progressive motility (P = 0.01). No statistically significant differences in semen parameters were found between smokers and non-smokers or between males who consumed alcohol versus those that did not. However, stress had a negative effect on sperm concentration. In conclusion, the prevalence of normal semen parameters in the study population evaluated was markedly lower than that reported for the general population (60-70%). Since sperm concentration did not appear to decrease with age, the decrease observed in the last decades may be related to exposure to environmental toxicants during the fetal period, as previously suggested.

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