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Updated: May 29, 2026

Sperm Collection of Differential Quality Using Density Gradient Centrifugation
Published on: November 29, 2018
Elimination of bacteria from human semen during sperm preparation using density gradient centrifugation with a novel
J Fourie1, N Loskutoff, C Huyser
1Department of Obstetrics and Gynaecology, Steve Biko Academic Hospital, University of Pretoria, Pretoria, South Africa. jozef.fourie@up.ac.za
Abstract:
The occurrence of bacteria in sperm samples intended for in vitro fertilisation can compromise the outcome of assisted reproductive techniques. Effective semen processing procedures should therefore be implemented to remove bacteria from semen. Unfortunately, technique failure does occur whereby bacteria can be found in processed sperm preparations. To improve the effectiveness of semen processing, a novel centrifuge tube insert was developed to facilitate the layering of density gradients and semen, and to prohibit the re-infection of purified sperm pellets. The purpose of this study was to: (i) determine the prevalence and type of bacteria present in semen of patients participating in the Unit's assisted reproduction program and (ii) evaluate the effectiveness of density gradient centrifugation with the novel tube insert, for the elimination of bacteria and yeast from spiked human semen samples. A survey in 2007-2010 indicated that 50% of semen samples were found to have positive bacterial cultures. Semen processing by means of density gradient centrifugation with the novel tube insert eliminated significantly more in vitro derived (spiked) bacteria and yeast from semen compared to processing without the insert (P < 0.004). Therefore, it is highly recommended that the centrifuge tube insert, ProInsert™, be incorporated into assisted reproductive programs.
Insights
Bacterial contamination in sperm samples can harm assisted reproduction outcomes. A new centrifuge tube insert, ProInsert™, significantly improves bacterial and yeast removal during semen processing, enhancing fertility success rates.
Area of Science:
- Reproductive Biology
- Microbiology
Background:
- Bacterial presence in sperm samples can negatively impact assisted reproductive techniques (ART).
- Current semen processing methods may not always eliminate microbial contamination effectively.
- A novel centrifuge tube insert (ProInsert™) was developed to enhance semen processing and prevent re-infection.
Purpose of the Study:
- To determine the prevalence and types of bacteria in semen samples from an ART program.
- To evaluate the efficacy of density gradient centrifugation (DGC) with the ProInsert™ for eliminating bacteria and yeast from spiked human semen.
Main Methods:
- A survey of semen samples from 2007-2010 identified bacterial prevalence.
- In vitro experiments tested DGC with and without the ProInsert™ for removing spiked bacteria and yeast from semen.
Main Results:
- A 2007-2010 survey revealed that 50% of semen samples had positive bacterial cultures.
- DGC utilizing the ProInsert™ demonstrated significantly greater elimination of spiked bacteria and yeast compared to DGC alone (P < 0.004).
Conclusions:
- The ProInsert™ significantly enhances the removal of bacterial and yeast contaminants from human semen during processing.
- Incorporating the ProInsert™ into ART programs is recommended to improve the effectiveness of semen processing and ART outcomes.
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