Timing Is Critical: Post-Collection Sperm Dna Integrity and Motility

Objective: Sperm DNA fragmentation (SDF) is known to be detrimental to the quality of sperm, which can negatively impact assisted reproductive technology (ART) cycles. To evaluate the impact of time on sperm DNA integrity post-collection, we analyzed SDF in seminal fluid over time to simulate conditions of off-site semen collection. Design: Prospective experiment Materials and Methods: 14 normal semen samples were collected onsite and analyzed SDF using Halosperm G2 kit (Halotech, Spain). Following the vendor’s protocol, sperm with intact DNA showed dispersion halos, while sperm with fragmented DNA had absent or minimal halos. Positive and negative controls were included in each experiment. Semen samples were assessed at 0-, 1-, 2-, and 4-hour time points (0 hour is defined as 15 minutes after liquefaction period). Statistical analyses were performed using ANOVA with Tukey HSD test for group comparison. Data were normally distributed (p<0.05 considered significant), and equal variances were confirmed by Levene’s test. Results: Initial semen parameter results of the group yielded an average semen volume of 3.7mL, average concentration of 42.2 million/mL, average motility of 46.9%, average normal morphology of 4%, and negative white blood cells for all samples by qualitative test (Medical Electronic Systems, CA, USA). Average sperm motility was significantly reduced, and average SDF was significantly increased over each time point after semen collection (Table 1). This indicates that sperm DNA could not hold its integrity when the seminal fluid is stagnant for longer durations between ejaculation and sperm preparation. Conclusions: This decrease in sperm quality over time could be used to influence fertility clinics’ protocols when considering sperm preparation for ART cycles, especially when semen collection occurs off-site from the andrology lab for ART. Every lab should establish an acceptable time threshold for receiving semen from off-site collection in setting of IUI or IVF sperm preparation.