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Reduced bacterial load in stallion semen by modified single layer centrifugation or sperm washing.
Abstract: The presence of bacteria poses a significant challenge to the quality of stallion semen used in artificial insemination. The bacterial content of insemination doses arises from various sources, such as the healthy stallion, environment, and collection equipment, and is implicated in fertility problems as well as reduced sperm quality during storage. The conventional approach of adding antibiotics to semen extenders raises concerns about antimicrobial resistance and potential negative effects on sperm characteristics, and may not be effective in inhibiting all bacteria. The objective of this study was to determine whether an innovative alternative to antibiotic usage - centrifugation through a single layer of a low density colloid (SLC) - could reduce the bacterial load in stallion semen, and to compare sperm characteristics in samples arising from this procedure, or simple extension of the ejaculate in semen extender, or from sperm washing, i.e. adding extender and then centrifuging the sample to allow the removal of most of the seminal plasma and extender. Eighteen semen samples were collected from six stallions. The semen samples were split and extended prior to washing or SLC, or received no further treatment other than extension. After preparation aliquots from each type of sample were sent for bacteriological examination; the remaining samples were stored for up to 72 h, with daily checks on sperm quality. The low density colloid SLC outperformed sperm washing or extension for bacterial reduction, effectively removing several bacterial species. The bacterial load in the samples was as follows: extended semen, 16 ± 6.7 × 10; washed, 5.8 ± 2.0 × 10; SLC, 2.3 ± 0.88 × 10, p < 0.0001. In addition, SLC completely removed some bacterial species, such as Staphylococcus xylosus. Although there is no selection for robust spermatozoa with the low density colloid, sperm motility, membrane integrity, and DNA fragmentation were not different to washed sperm samples. These findings suggest that SLC with a low density colloid offers a promising method for reducing bacterial contamination in stallion semen without resorting to antibiotics.
Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.
Publication Date: 2023-12-29 PubMed ID: 38171197DOI: 10.1016/j.theriogenology.2023.12.034Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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The research reveals an effective strategy for reducing bacterial contamination in stallion semen for artificial insemination by using a method called the low-density single layer centrifugation (SLC), providing an alternative to antibiotics usage. The study also compared sperm quality in samples treated with the SLC method with those extended in semen extender or a simple sperm washing process.
Research Methodology
- This study utilized eighteen semen samples collected from six stallions.
- The semen samples were divided and prepared either by washing or SLC, or they were extended with no additional treatments.
- Post preparation, some samples were dispatched for bacteriological examination.
- The remaining samples were preserved up to 72 hours with daily assessments on sperm quality.
Study Findings
- The results showed that the SLC methodology, using a low-density colloid, was more effective in decreasing bacterial content in the semen samples than either sperm washing or semen extension.
- The bacterial counts in the samples were found to be lowest in the SLC processed samples, followed by those treated with washing and extension. The load in the extended semen was reported as 16 ± 6.7 × 10, while for washed semen 5.8 ± 2.0 × 10, and for SLC, it was the lowest at 2.3 ± 0.88 × 10.
- Furthermore, the SLC method was found to remove completely certain bacterial species, such as Staphylococcus xylosus.
- The study reported no significant difference in sperm quality (motility, membrane integrity, and DNA fragmentation) between washed and SLC treated samples, indicating that the low-density colloid used in SLC does not select for robust spermatozoa.
Summary and Implication
- The study demonstrates that the low-density single layer centrifugation technique provides an effective and promising option for reducing bacterial contamination in stallion semen for artificial insemination, without resorting to antibiotics.
- This could be a significant strategy to minimize the creation and spread of antibiotic-resistant bacterial strains.
Cite This Article
APA
Malaluang P, Wagner LH, Cojkic A, Spergser J, Aurich C, Morrell JM.
(2023).
Reduced bacterial load in stallion semen by modified single layer centrifugation or sperm washing.
Theriogenology, 216, 111-117.
https://doi.org/10.1016/j.theriogenology.2023.12.034 Publication
Researcher Affiliations
- Department of Clinical Sciences, Swedish University of Agricultural Sciences (SLU), 75007, Uppsala, Sweden; Faculty of Veterinary Sciences, Mahasarakham University, Maha Sarakham, 40000, Thailand.
- Artificial Insemination and Embryo Transfer, Department for Small Animals and Horses, University of Veterinary Medicine, Vienna, Austria.
- Department of Clinical Sciences, Swedish University of Agricultural Sciences (SLU), 75007, Uppsala, Sweden.
- Institute of Microbiology, Department of Pathobiology, University of Veterinary Medicine, Vienna, Austria.
- Artificial Insemination and Embryo Transfer, Department for Small Animals and Horses, University of Veterinary Medicine, Vienna, Austria.
- Department of Clinical Sciences, Swedish University of Agricultural Sciences (SLU), 75007, Uppsala, Sweden. Electronic address: jane.morrell@slu.se.
Conflict of Interest Statement
Declaration of competing interest JMM is the inventor and one of the patent holders of the colloid used in this study.
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