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Bacterial contamination and sperm recovery after semen preparation by density gradient centrifugation using
C M Nicholson1, L Abramsson, S E Holm
1Department of Obstetrics and Gynecology, Umeå University Hospital, S-90185 Umeå, Sweden.
Abstract:
The effects of density gradient centrifugation through silane-coated silica particles (PureSperm) using 100, 200, 300 and 500 g on bacterial contamination of sperm samples and recovery of motile spermatozoa from sperm samples were investigated with conventional culturing techniques and microscopic visual assessment. The recovery of motile spermatozoa was variable and was not improved using 500 g compared to the recommended 300 g. The bacterial contamination was highly decreased by gradient centrifugation through PureSperm and was almost abolished when strict aseptic techniques were used, with changes to sterile Pasteur pipettes and tubes prior to washing procedures.
Insights
Density gradient centrifugation using PureSperm significantly reduces bacterial contamination in sperm samples. Optimal sperm recovery was achieved at 300 g, with higher forces not improving results.
Area of Science:
- Reproductive biology
- Microbiology
- Andrology
Background:
- Bacterial contamination poses a significant challenge in sperm processing.
- Effective sperm washing techniques are crucial for assisted reproductive technologies.
Purpose of the Study:
- To evaluate the impact of density gradient centrifugation with silane-coated silica particles (PureSperm) on bacterial contamination.
- To assess the recovery of motile spermatozoa at different centrifugation speeds.
Main Methods:
- Sperm samples were subjected to density gradient centrifugation using PureSperm at 100, 200, 300, and 500 g.
- Bacterial contamination was assessed using conventional culturing techniques.
- Motile spermatozoa recovery was evaluated via microscopic visual assessment.
Main Results:
- Density gradient centrifugation through PureSperm markedly decreased bacterial contamination.
- Bacterial contamination was nearly eliminated with the implementation of strict aseptic techniques.
- Motile spermatozoa recovery varied and was not enhanced by using 500 g compared to the standard 300 g.
Conclusions:
- Density gradient centrifugation with PureSperm is effective in reducing bacterial contamination in sperm samples.
- Strict aseptic techniques are essential for almost abolishing bacterial contamination.
- Centrifugation at 300 g is sufficient for optimal motile spermatozoa recovery.