Bacterial contamination and sperm recovery after semen preparation by density gradient centrifugation using silane-coated silica particles at different g forces

Nicholson, Cheryl; Abramsson, Leif; Holm, Sonya; Bjurulf, E.

doi:10.1093/humrep/15.3.662

Cited by 47 publications

(36 citation statements)

References 29 publications

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“…The bacteria may have contributed to death of spermatozoa through the production of endotoxins. Colloidal centrifugation has been reported to remove bacteria from human semen samples [18] and may have contributed to the increased survival of the boar spermatozoa seen in the current study. It might be possible to reduce the use of antibiotics in semen extenders if SLC is employed to process the spermatozoa, thus reducing the environmental impact of these antimicrobials.…”

Section: Discussionmentioning

confidence: 81%

Selection of Boar Spermatozoa Using Centrifugation on a Glycidoxypropyltrimethoxysilane-Coated Silica Colloid

Morrell

Saravia

Wienen

et al. 2009

Journal of Reproduction and Development

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Abstract. Use of sperm separation methods such as density gradient centrifugation for selecting the best spermatozoa for animal breeding is constrained by the problem of dealing with the large volumes of ejaculate produced by the males of some species, such as boars. The purpose of this study was to compare density gradient centrifugation (DGC) with centrifugation on a single layer of colloid (SLC) for the preparation of ejaculated boar spermatozoa using Androcoll TM -P. There was no difference between the two techniques in terms of sperm motility or duration of motility after selection, and sperm motility was retained for at least 24 h longer in the centrifuged sperm preparations than in controls (uncentrifuged aliquots). Sperm motility was significantly better (P<0.001) in the centrifuged sperm preparations (means ± sd: SLC 79.6 ± 8.1% and DGC 74.2 ± 12.0%) than in the uncentrifuged controls (62.9 ± 12.7%). The mean yield of motile spermatozoa for SLC was 67.5 ± 25.6%, and for DGC it was 59.6% ± 22.3% (not significant, ns). Sperm survival was significantly increased by colloid centrifugation (control 3.1 ± 0.3 days, SLC 5.5 ± 0.79 days, DGC 5.75 ± 0.62 days; P<0.001 for uncentrifuged versus centrifuged; SLC vs. DGC, ns). Moreover, boar spermatozoa could be stored for 24 h before centrifugation without any detrimental effect on sperm motility or duration of motility. In a further experiment, larger volumes of ejaculate were processed easily on a modified SLC, indicating that this method may be practical for processing large volumes of boar ejaculates. Key words: Boar spermatozoa, Colloids, Density gradient centrifugation, Single layer centrifugation (J. Reprod. Dev. 55: [547][548][549][550][551][552] 2009) ecently, density gradient centrifugation (DGC) has been suggested as a means of selecting animal spermatozoa for artificial breeding [1][2][3]. It is one of the methods recommended by the World Health Organisation for preparing human spermatozoa for assisted reproduction [4]. DGC has been used to prepare bovine spermatozoa for in vitro fertilization [5] and to prepare stallion spermatozoa for research purposes [6,7]. However, widespread application of the DGC-technique in processing animal spermatozoa has been hampered by a lack of animal-specific colloid formulations and the difficulty of processing sufficient spermatozoa for the large insemination doses required in domestic animals. Percoll TM DGC has been used previously to prepare animal spermatozoa for IVF, but the manufacturer (Amersham Biosciences, Uppsala) withdrew the product from the market for clinical use in 1996, reportedly because the polyvinylpropylene (PVP) it contains may damage spermatozoa [8,9].Recently, colloid formulations for use with animal spermatozoa, based on silane-coated silica, have been developed at the Swedish University of Agricultural Sciences (SLU) [10]. Preliminary studies with stallion spermatozoa have shown that the new colloid formulation could be used successfully as a single colloid layer for processing stallion sper...

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Section: Discussionmentioning

confidence: 81%

Selection of Boar Spermatozoa Using Centrifugation on a Glycidoxypropyltrimethoxysilane-Coated Silica Colloid

Morrell

Saravia

Wienen

et al. 2009

Journal of Reproduction and Development

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“…Sperm washing was carried out using density gradient centrifugation, as described previously (15). Briefly, 1 mL of liquefied semen was placed on the top of two 1-mL layers of 80% and 40% (vol/vol) of Cook Sperm Gradient Medium (Cook, Brisbane, Australia) in a sterile conical tube (Becton Dickinson Co., Oxford, United Kingdom).…”

Section: Sperm Washing Proceduresmentioning

confidence: 99%

“…The clinical importance of sperm washing is evident as bacteriospermia in the male partners of couples attending for ART is common (11). However, although it has been suggested that sperm washing was highly effective in removing bacteria (12), later studies (13)(14)(15) have suggested it may only be 95% efficient. In addition, since some of these studies were carried out, it is now known that there is a tendency of some bacteria (e.g., Escherichia coli) to adhere to spermatozoa (16).…”

mentioning

confidence: 99%

Real-time polymerase chain reaction shows that density centrifugation does not always remove Chlamydia trachomatis from human semen

Al-Mously

Cross

Eley

et al. 2009

Fertility and Sterility

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“…The last line of defence against seminal-derived bacterial contamination of the embryo culture system, is semen processing, utilizing strict aseptic techniques and proper changing of sterile pipette tips and tubes between the DGC and washing procedures (Nicholson et al, 2000).…”

Section: Discussion Presence Of Bacteria In Semenmentioning

confidence: 99%

Elimination of bacteria from human semen during sperm preparation using density gradient centrifugation with a novel tube insert

Fourie

Loskutoff²,

Huyser

2011

Andrologia

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SummaryThe occurrence of bacteria in sperm samples intended for in vitro fertilization, 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 reinfection of purified sperm pellets. The purpose of this study was to: 1) determine the Therefore, it is highly recommended that the centrifuge tube insert, ProInsert TM , be incorporated into assisted reproductive programs.3

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Bacterial contamination and sperm recovery after semen preparation by density gradient centrifugation using silane-coated silica particles at different g forces

Cited by 47 publications

References 29 publications

Selection of Boar Spermatozoa Using Centrifugation on a Glycidoxypropyltrimethoxysilane-Coated Silica Colloid

Selection of Boar Spermatozoa Using Centrifugation on a Glycidoxypropyltrimethoxysilane-Coated Silica Colloid

Real-time polymerase chain reaction shows that density centrifugation does not always remove Chlamydia trachomatis from human semen

Elimination of bacteria from human semen during sperm preparation using density gradient centrifugation with a novel tube insert

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