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

Fourie, Jozef Markus; Loskutoff, N.M.; Huyser, Carin

doi:10.1111/j.1439-0272.2011.01217.x

Cited by 22 publications

(14 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…[1415] The presence of bacteria in semen may be an early warning signal for altered reproductive potential of men. [5] The harmful effects of bacteria on spermatozoa may depend on the type and species of microorganisms invading, colonizing, or infecting the male genital tract. [16] Antibiotic treatment of leukocytospermic men without diagnosed genital tract infections reported a significant improvement of ejaculate quality.…”

Section: Discussionmentioning

confidence: 99%

“…[3] Nicholson et al have reported almost bacteria abolished samples using density gradients method with strict aseptic conditions. [4] Samples with significantly lower bacterial counts were obtained with modified density gradients centrifugation using an inner tube insertion compared with the conventional method by Fourie et al [5] A similar method employed an inner column of Percoll has been reported by Kaneko et al in 1986 with a greater success. [6] Sperm preparation methods are not guaranteed options for complete elimination of bacteria.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Antibiotics supplemented culture media can eliminate non-specific bacteria from human semen during sperm preparation for intra uterine insemination

et al. 2014

View full text Add to dashboard Cite

RATIONALE:Bacterial flora can be isolated from many semen samples of subfertile males. Bacteriospermia can compromise the outcome of intra uterine insemination (IUI) by contaminating the post-processed sperm sample.OBJECTIVES:The objective of the present study is to determine the efficacy of penicillin and streptomycin in eliminating the bacteria from semen samples in the sperm processing procedure, and to assess the effects of antibiotics on sperm motility, survivability, and pregnancy rates.DESIGN AND SETTINGS:A prospectively controlled study was carried out using couples undergoing IUI with their informed consent.INTERVENTION:Sperm processing using the swim-up technique in penicillin and streptomycin supplemented culture medium.SUBJECTS AND METHODS:Couples were consecutively allocated in two groups for sperm processing (a) Group AB+ (antibiotics supplemented culture medium, n = 33) and (b) Group AB− (antibiotic free culture medium, n = 33). Semen culture was performed before and after sperm processing. Sperm motility was assessed immediately after processing and after 24 h of incubation.RESULTS:Bacterial isolates were found in 20 (60.6%) and 22 (66.1%) of samples before processing in Groups AB+ and AB− respectively. Addition of antibiotics resulted in completely eliminating non-specific bacteria from semen samples without affecting sperm motility. In vitro survival rate of sperm enhanced in AB+ group compared with AB− group (motile sperm after 24 h), 62.21% (standard deviation [SD]: 37.27) versus 41.36% (SD: 30.78), P = 0.012. Pregnancy rate, was comparable between two groups (9% in Group AB+ vs. 6% in Group AB−, P = 0.45).CONCLUSION:Penicillin streptomycin combination could completely eliminate non-specific bacteria from semen samples during sperm processing in this population. The types of antibiotics and dosage used did not seem to have any harmful effects on human sperm.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Antibiotics supplemented culture media can eliminate non-specific bacteria from human semen during sperm preparation for intra uterine insemination

et al. 2014

View full text Add to dashboard Cite

show abstract

“…The addition of rhProtease to density layers in combination with the usage of a ProInsert™ (Nidacon, International), could therefore be beneficial, specifically when assisting patients with potentially infected semen samples [34]. However, the effect rhProtease has on sperm parameters should be considered prior to its inclusion into DGC protocols.…”

Section: Discussionmentioning

confidence: 99%

Treatment of human sperm with serine protease during density gradient centrifugation

Fourie

Loskutoff²,

Huyser

2012

J Assist Reprod Genet

Self Cite

View full text Add to dashboard Cite

Purpose Seminal pathogens can bind specifically or nonspecifically to spermatozoa, rendering semen decontamination procedures ineffective, whereby vertical or horizontal transmission of the infection could occur. Serine proteases have been demonstrated to effectively inactivate viruses and to break pathogen-sperm bonds. However, the addition of a protease to density gradient layers during semen processing could negatively impact on sperm parameters. This study investigated the effect of the addition of a recombinant, human-sequence protease (rhProtease) on sperm parameters during density gradient centrifugation. Methods (i) Pooled semen samples (n09) were split and processed by density gradient centrifugation, with the top density layers supplemented, or non-supplemented with rhProtease at three different concentrations (diluted 2, 10 and 20 times). Sperm parameters were then analysed by flow cytometry and computer-assisted semen analyses.(ii) Semen samples (n05) were split and similarly processed using PureSperm® Pro, with rhProtease in the 40 % density gradient layer, or standard PureSperm® not supplemented with rhProtease (Nidacon, International) respectively. The Hemizona assay was then utilized to compare sperm-zona binding post processing.Results Evaluation of sperm parameters indicated that rhProtease did not, at any of the tested concentrations, have an impact on (i) mitochondrial membrane potential, vitality, motility, or (ii) zona binding potential. Conclusion We report that the addition of rhProtease to density gradients is a non-detrimental approach that could improve the effectiveness of semen processing for the elimination of seminal pathogens, and benefit assisted reproduction outcome.

show abstract

“…Semen decontamination procedures employed at RBL consist of semen processing by discontinuous density gradient centrifugation in combination with the ProInsert™. This method of semen decontamination has previously been reported to be superior than standard discontinues density gradient centrifugation in the removal of in vitro added bacteria from semen (Fourie et al, 2012). The removal of HIV-1 from semen should therefore also be improved by utilization of the decontamination technique.…”

Section: Discussionmentioning

confidence: 99%

Semen decontamination for the elimination of seminal HIV-1

Fourie

Loskutoff²,

Huyser

2015

Reproductive BioMedicine Online

View full text Add to dashboard Cite

The risk of HIV transmission to the female partner, or potential offspring of an HIV-1 infected male can be reduced using semen decontamination procedures prior to assisted reproductive treatment (ART). The objective of this study was to determine the efficiency to decontaminate semen samples (N=186) from 95 HIV-1 sero-positive patients. Aliquots of neat semen were submitted for viral validation by qualitative and quantitative polymerase chain reaction. Semen samples were processed by density gradient centrifugation in combination with a ProInsert™ tube where after aliquots of the processed sperm samples were analysed for the presence of HIV-1. Fifty four percent of all tested neat semen samples tested positive for HIV-1 DNA, RNA or both (13.4%, 11.3% and 29.0%, respectively). From a total of 103 processed sperm samples that were submitted for viral validation, two samples tested positive for HIV-1 DNA and none for RNA. In conclusion, semen processing with the ProInsert™ followed by viral validation of processed sperm samples should be performed when providing assisted reproductive treatment to couples where the male partner is HIV-1 seropositive.

show abstract

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

Cited by 22 publications

References 26 publications

Antibiotics supplemented culture media can eliminate non-specific bacteria from human semen during sperm preparation for intra uterine insemination

Antibiotics supplemented culture media can eliminate non-specific bacteria from human semen during sperm preparation for intra uterine insemination

Treatment of human sperm with serine protease during density gradient centrifugation

Semen decontamination for the elimination of seminal HIV-1

Contact Info

Product

Resources

About