Article Data

  • Views 503
  • Dowloads 130

Original Research

Open Access

Elimination of bacteria from semen using a combination of density gradient centrifugation and swim-up

  • Zhiheng Chen1,†,
  • Lingxiao Jiang2,†,
  • Hui Wang11
  • Wumin Quan1
  • Li Yang1
  • Chunquan Ou3,*,
  • Ling Sun1,*,

1Center of Reproductive Medicine, Guangzhou Women and Children’s Medical Center, Guangzhou Medical University, 510140 Guangzhou, Guangdong, China

2Clinical Laboratory, Zhujiang Hospital, Southern Medical University, 510280 Guangzhou, Guangdong, China

3State Key Laboratory of Organ Failure Research, Department of Biostatistics, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, 510515 Guangzhou, Guangdong, China

DOI: 10.31083/j.jomh1806124 Vol.18,Issue 6,June 2022 pp.1-6

Submitted: 12 August 2021 Accepted: 03 October 2021

Published: 30 June 2022

*Corresponding Author(s): Chunquan Ou E-mail: ouchunquan@hotmail.com
*Corresponding Author(s): Ling Sun E-mail: sunling6299@163.com

† These authors contributed equally.

Abstract

Background: To evaluate the efficiency of bacterial clearance of different sperm preparation procedure. Methods: A total of 46 semen samples were collected in this study, each semen sample was divided into four equal parts (0.5 mL each). The first part was prepared by density-gradient centrifugation (D-group), the second part was prepared by swim-up (S-group), the third part was treated with density-gradient centrifugation combined with swim-up (D+S-group), and the fourth group was the original raw semen group (O-group) regarded as the control. After each semen preparation procedure, the O-group and suspensions were inoculated and incubated for bacterial identification and colony counting. Results: Initially only 8.7% (4/46) of samples were bacteria free. After processing, the bacterial clearance rates were 23.8% for D-group, 57.4% for S-group and 97.8% for D+S-group (p < 0.001). Multiple bacterial strains were observed in 37 samples with 117 different bacterial strains in all identified in the original semen. 44.4% of the staphylococci and 89.7% of streptococci were not eliminated in D-group. In S-group, the corresponding rates were 24.4% and 35.9%. In D+S-group there were no remaining strains of staphylococci and only 2.6% of streptococci remained. After the combined procedure, the number of bacterial colonies fell dramatically after processing. Conclusion: The combined D+S protocol appears to be substantially more efficient than either method alone in eliminating bacteria from semen samples.


Keywords

Semen; Bacteria; Density-gradient centrifugation; Swim-up


Cite and Share

Zhiheng Chen,Lingxiao Jiang,Hui Wang1,Wumin Quan,Li Yang,Chunquan Ou,Ling Sun. Elimination of bacteria from semen using a combination of density gradient centrifugation and swim-up. Journal of Men's Health. 2022. 18(6);1-6.

References

[1] Zhu GJ, Wei YL, Hu J, Liu Q. Microorganism contamination in in vitro fertilization-embryo transfer system and their sources. Zhonghua Fu Chan Ke Za Zhi. 2004. 39: 382–384.

[2] Ben-Chetrit A, Shen O, Haran E, Brooks B, Geva-Eldar T, Mar-galioth EJ. Transfer of embryos from yeast-colonized dishes. Fertility and Sterility. 1996; 66: 335–337.

[3] Cottell E, McMorrow J, Lennon B, Fawsy M, Cafferkey M, Har-rison RF. Microbial contamination in an in vitro fertilization-embryo transfer system. Fertility and Sterility. 1996; 66: 776–780.

[4] Kastrop PMM, de Graaf-Miltenburg LAM, Gutknecht DR, Weima SM. Microbial contamination of embryo cultures in an ART laboratory: sources and management. Human Reproduc-tion. 2007; 22: 2243–2248.

[5] Mitchell JG, Kogure K. Bacterial motility: links to the environ-ment and a driving force for microbial physics. FEMS Microbi-ology Ecology. 2006; 55: 3–16.

[6] Megory E, Zuckerman H, Shoham Z, Lunenfeld B. Infections and male fertility. Obstetrical and Gynecological Survey. 1987; 42: 283–290.

[7] Domes T, Lo KC, Grober ED, Mullen JBM, Mazzulli T, Jarvi K. The incidence and effect of bacteriospermia and elevated seminal leukocytes on semen parameters. Fertility and Sterility. 2012; 97: 1050–1055.

[8] Rusz A, Pilatz A, Wagenlehner F, Linn T, Diemer T, Schuppe HC, et al. Influence of urogenital infections and inflammation on semen quality and male fertility. World Journal of Urology. 2012; 30: 23–30.

[9] Bielanski A. Disinfection procedures for controlling microor-ganisms in the semen and embryos of humans and farm animals. Theriogenology. 2007; 68: 1–22.

[10] Sacks PC, Simon JA. Infectious complications of intrauterine insemination: a case report and literature review. International Journal of Fertility. 1992; 36: 331–339.

[11] Fourie J, Loskutoff N, Huyser C. Elimination of bacteria from human semen during sperm preparation using density gradient centrifugation with a novel tube insert. Andrologia. 2012; 44: 513–517.

[12] Kim FY, Goldstein M. Antibacterial skin preparation decreases the incidence of false-positive semen culture results. The Journal of Urology. 1999; 161: 819–821.

[13] Krissi H, Orvieto R, Ashkenazi J, Gilboa Y, Shalev J, Moscov-itch I, et al. Effect of contaminated preprocessed semen on fer-tilization rate and embryo quality in assisted reproductive tech-niques. Gynecological Endocrinology. 2004; 18: 63–67.

[14] World Health Organization. WHO laboratory manual for the Examination and processing of human semen. 2010. Avail-able at: https://apps.who.int/iris/bitstream/handle/10665/44261/ 9789750011245_tur.pdf (Accessed: 9 October 2021).

[15] Elder K, Baker DJ, Ribes JA. Infections, infertility and assisted reproduction. Cambridge University Press: Cambridge. 2005.

[16] Magli MC, Gianaroli L, Fiorentino A, Ferraretti AP, Fortini D, Panzella S. Improved cleavage rate of human embryos cul-tured in antibiotic-free medium. Human Reproduction. 1996; 11: 1520–1524.

[17] Catry B, Van Duijkeren E, Pomba MC, Greko C, Moreno MA, Pyorala S, et al. Reflection paper on MRSA in food-producing and companion animals: epidemiology and control options for human and animal health. Epidemiology and Infection. 2010; 138: 626–644.

[18] Duijkeren EV, Box ATA, Schellen P, Houwers DJ, Fluit AC. Class 1 integrons in Enterobacteriaceae isolated from clinical in-fections of horses and dogs in the Netherlands. Microbial Drug Resistance. 2006; 11: 383–386.

[19] Martínez-Pastor F, Lacalle E, Martínez-Martínez S, Fernández-Alegre E, Álvarez-Fernández L, Martinez-Alborcia MJ, et al. Low density Porcicoll separates spermatozoa from bacteria and retains sperm quality. Theriogenology. 2021; 165: 28–36.

[20] Morrell JM, Wallgren M. Removal of bacteria from boar ejacu-lates by Single Layer Centrifugation can reduce the use of antibi-otics in semen extenders. Animal Reproduction Science. 2011; 123: 64–69.

[21] Nicholson CM, Abramsson L, Holm SE, Bjurulf E. Bacterial contamination and sperm recovery after semen preparation by density gradient centrifugation using silane-coated silica parti-cles at different g forces. Human Reproduction. 2000; 15: 662–666.

[22] Marchesi DE, Qiao J, Feng HL. Embryo manipulation and im-printing. Seminars in Reproductive Medicine. 2012; 30: 323–334.

[23] Henkel RR, Schill WB. Sperm preparation for ART. Reproduc-tive Biology and Endocrinology. 2003; 1: 1–22.

[24] Knox CL, Allan JA, Allan JM, Edirisinghe WR, Stenzel D, Lawrence FA, et al. Ureaplasma parvum and Ureaplasma ure-alyticum are detected in semen after washing before assisted re-productive technology procedures. Fertility and Sterility. 2003; 80: 921–929.

[25] Abeysundara PK, Dissanayake D, Wijesinghe PS, Perera R, Nishad A. Efficacy of two sperm preparation techniques in re-ducing non-specific bacterial species from human semen. Jour-nal of Human Reproductive Sciences. 2013; 6: 152–157.

[26] Huyser C, Fourie FLR, Oosthuizen M, Neethling A. Microbial flora in semen during in vitro fertilization. Journal of in Vitro Fertilization and Embryo Transfer. 1991; 8: 260–264.

[27] Al-Mously N, Cross NA, Eley A, Pacey AA. Real-time poly-merase chain reaction shows that density centrifugation does not always remove Chlamydia trachomatis from human semen. Fer-tility and Sterility. 2009; 92: 1606–1615.

[28] Kuzan FB, Hillier SL, Zarutskie PW. Comparison of three wash techniques for the removal of microorganisms from semen. Ob-stetrics and Gynecology. 1988; 70: 836–839.

[29] Karlstrom PO, Hjelm E, Lundkvist O. Comparison of the ability of two sperm preparation techniques to remove microbes. Hu-man Reproduction. 1991; 6: 386–389.

[30] Morrell JM, Klein C, Lundeheim N, Erol E, Troedsson MHT. Removal of bacteria from stallion semen by colloid centrifuga-tion. Animal Reproduction Science. 2014; 145: 47–53.

[31] Guimarães T, Lopes G, Pinto M, Silva E, Miranda C, Correia MJ, et al. Colloid centrifugation of fresh stallion semen be-fore cryopreservation decreased microorganism load of frozen-thawed semen without affecting seminal kinetics. Theriogenol-ogy. 2015; 83: 186–191.

[32] Ahmad L, Jalali S, Shami SA, Akram Z. Sperm preparation: DNA damage by comet assay in normo- and teratozoospermics. Archives of Andrology. 2008; 53: 325–338.

[33] Diemer T, Weidner W, Michelmann HW, Schiefer HG, Rovan E, Mayer F. Influence of Escherichia coli on motility parameters of human spermatozoa in vitro. International Journal of Andrology. 1996; 19: 271–277.


Abstracted / indexed in

Science Citation Index Expanded (SciSearch) Created as SCI in 1964, Science Citation Index Expanded now indexes over 9,200 of the world’s most impactful journals across 178 scientific disciplines. More than 53 million records and 1.18 billion cited references date back from 1900 to present.

Journal Citation Reports/Science Edition Journal Citation Reports/Science Edition aims to evaluate a journal’s value from multiple perspectives including the journal impact factor, descriptive data about a journal’s open access content as well as contributing authors, and provide readers a transparent and publisher-neutral data & statistics information about the journal.

Directory of Open Access Journals (DOAJ) DOAJ is a unique and extensive index of diverse open access journals from around the world, driven by a growing community, committed to ensuring quality content is freely available online for everyone.

SCImago The SCImago Journal & Country Rank is a publicly available portal that includes the journals and country scientific indicators developed from the information contained in the Scopus® database (Elsevier B.V.)

Publication Forum - JUFO (Federation of Finnish Learned Societies) Publication Forum is a classification of publication channels created by the Finnish scientific community to support the quality assessment of academic research.

Scopus: CiteScore 0.7 (2022) Scopus is Elsevier's abstract and citation database launched in 2004. Scopus covers nearly 36,377 titles (22,794 active titles and 13,583 Inactive titles) from approximately 11,678 publishers, of which 34,346 are peer-reviewed journals in top-level subject fields: life sciences, social sciences, physical sciences and health sciences.

Norwegian Register for Scientific Journals, Series and Publishers Search for publication channels (journals, series and publishers) in the Norwegian Register for Scientific Journals, Series and Publishers to see if they are considered as scientific. (https://kanalregister.hkdir.no/publiseringskanaler/Forside).

Submission Turnaround Time

Conferences

Top