Colloid centrifugation of fresh stallion semen before cryopreservation decreased microorganism load of frozen-thawed semen without affecting seminal kinetics.
Abstract: Freezability of equine semen may be influenced by microorganism population of semen. The objective of this study was to verify the effect of single-layer density gradient centrifugation (SLC) of fresh semen before cryopreservation on semen's microbial load (ML) and sperm cells kinetics after freezing-thawing. For that, one ejaculate was collected from 20 healthy stallions and split into control (C) samples (cryopreserved without previous SLC) and SLC samples (subjected to SLC). Semen cryopreservation was performed according to the same protocol in both groups. Microbial load of each microorganism species and total microbial load (TML) expressed in colony-forming units (CFU/mL) as well as frozen-thawed sperm kinetics were assessed in both groups. Additional analysis of the TML was performed, subdividing the frozen-thawed samples in "suitable" (total motility ≥ 30%) and "unsuitable" (total motility < 30%) semen for freezing programs, and comparing the C and SLC groups within these subpopulations. After thawing, SLC samples had less (P < 0.05) TML (88.65 × 10(2) ± 83.8 × 10(2) CFU/mL) than C samples (155.69 × 10(2) ± 48.85 × 10(2) CFU/mL), mainly due to a reduction of Enterococcus spp. and Bacillus spp. A relationship between post-thaw motility and SLC effect on ML was noted, as only in samples with more than 30% total motility was ML reduced (P < 0.05) by SLC (from 51.33 × 10(2) ± 33.26 × 10(2) CFU/mL to 26.68 × 10(2) ± 12.39 × 10(2) CFU/mL in "suitable" frozen-thawed semen vs. 240.90 × 10(2) ± 498.20 × 10(2) to 139.30 × 10(2) ± 290.30 × 10(2) CFU/mL in "unsuitable" frozen-thawed semen). The effect of SLC on kinetics of frozen-thawed sperm cells was negligible.
Copyright © 2015 Elsevier Inc. All rights reserved.
Publication Date: 2014-09-16 PubMed ID: 25287189DOI: 10.1016/j.theriogenology.2014.09.003Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research studied the effects of centrifugation on stallion semen prior to freezing it, indicating that this procedure effectively reduces microorganisms in semen without negatively impacting sperm cell movement.
Objective and Methodology
- The research aimed to assess the impact of single-layer density gradient centrifugation (SLC) on fresh stallion semen before freezing it. The specific focus was on how SLC influences the microbial load (ML) and sperm cell kinetics post the freezing-thawing process.
- Ejaculates were collected from 20 healthy stallions. These were split into two categories – control (C), where semen was cryopreserved without SLC, and SLC samples which were subjected to SLC.
- Upon undergoing the freezing-thawing procedure, both sets of samples were analyzed in terms of total microbial load (TML) and sperm cell kinetics. TML was estimated by measuring the concentration of colony-forming units (CFU/mL).
- Further analysis was conducted on samples grouped as “suitable” for freezing programs (those with total motility ≥ 30%) and “unsuitable” (those with total motility < 30%), classifying and comparing the control and SLC-groups within these.
Findings
- After thawing, SLC samples showed significantly lower TML than control samples. The decrease was primarily due to a reduction in the presence of bacteria like Enterococcus spp. and Bacillus spp.
- A correlation was observed between post-thaw motility and SLC effect on microorganism load. A reduction in microbial load due to SLC was noted only in samples that retained over 30% total motility after thawing.
- The SLC process had minimal effect on the movement of frozen-thawed sperm cells, suggesting that it does not significantly alter the sperm’s kinetic properties.
Implication
- This study implies that the single-layer density gradient centrifugation of fresh stallion semen prior to cryopreservation can effectively reduce the bacterial population without compromising the movement potential of the frozen-thawed sperm cells. This could potentially improve the safety and effectiveness of frozen semen in veterinary reproductive programs.
Cite This Article
APA
Guimarães T, Lopes G, Pinto M, Silva E, Miranda C, Correia MJ, Damásio L, Thompson G, Rocha A.
(2014).
Colloid centrifugation of fresh stallion semen before cryopreservation decreased microorganism load of frozen-thawed semen without affecting seminal kinetics.
Theriogenology, 83(2), 186-191.
https://doi.org/10.1016/j.theriogenology.2014.09.003 Publication
Researcher Affiliations
- Instituto de Ciências Biomédicas Abel Salazar, University of Porto, Portugal; Centro de Estudos de Ciência Animal (CECA/ICETA), University of Porto, Porto, Portugal.
- Instituto de Ciências Biomédicas Abel Salazar, University of Porto, Portugal; Centro de Estudos de Ciência Animal (CECA/ICETA), University of Porto, Porto, Portugal.
- Instituto de Ciências Biomédicas Abel Salazar, University of Porto, Portugal.
- Instituto de Ciências Biomédicas Abel Salazar, University of Porto, Portugal; Centro de Investigação em Biodiversidade e Recursos Genéticos (CIBIO), University of Porto, Porto, Portugal.
- Instituto de Ciências Biomédicas Abel Salazar, University of Porto, Portugal.
- Fundação Alter Real, Alter do Chão, Portugal.
- Private Practitioner, Évora, Portugal.
- Instituto de Ciências Biomédicas Abel Salazar, University of Porto, Portugal; Centro de Investigação em Biodiversidade e Recursos Genéticos (CIBIO), University of Porto, Porto, Portugal.
- Instituto de Ciências Biomédicas Abel Salazar, University of Porto, Portugal; Centro de Estudos de Ciência Animal (CECA/ICETA), University of Porto, Porto, Portugal. Electronic address: arocha@mail.icav.up.pt.
MeSH Terms
- Animals
- Bacteria / isolation & purification
- Centrifugation, Density Gradient / methods
- Centrifugation, Density Gradient / veterinary
- Colloids
- Colony Count, Microbial
- Cryopreservation / methods
- Cryopreservation / veterinary
- Horses
- Kinetics
- Male
- Semen / microbiology
- Semen Preservation / methods
- Semen Preservation / veterinary
- Sperm Motility / physiology
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