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Immediate and delayed (after cooling) effects of centrifugation on equine sperm.
Abstract: The objectives of this study were to determine the effects of centrifugation on equine sperm total and progressive motility, viability, and acrosomal integrity. We hypothesized that although high centrifugation forces would be detrimental to equine Equus caballus sperm, recovery rates would increase. Ejaculates from six stallions were collected, extended to a concentration of 25x10(6) cells/mL, and subjected for 10min to (1) no centrifugation (NC) or (2) centrifugation at 400xg, (3) 900xg, or (4) 4500xg. Before and after centrifugation (Day 0), and after 24h of cooling (Day 1), sperm motility was assessed by computer-assisted semen analysis, and samples were stained with SYBR-14/propidium iodide (PI) for viability and with PI/fluorescein isothiocyanate (FITC)-Peanut aglutinin (PNA) (Arachis hypogaea) for acrosomal integrity. The effect of treatment and day on motility, viability, and acrosomal integrity was determined using a mixed linear model. Compared with the other treatments, centrifugation at 4500xg reduced all end points measured (P<0.05). Both 400xg and 900xg yielded lower recovery rates than that of 4500xg (NC=100.0+/-0.0%; 400xg=54.4+/-8.6%; 900xg=75.0+/-7.1%; 4500xg=97.9+/-2.8%; P<0.05). Centrifugation at 400xg or 900xg did not damage equine sperm. Based on these findings, further studies of centrifugal forces between 900xg and 4500xg are warranted to determine the optimal force that maximizes recovery rate, minimizes sperm damage, and does not affect fertility.
Publication Date: 2009-11-17 PubMed ID: 19913898DOI: 10.1016/j.theriogenology.2009.09.003Google 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
- Research Support
- Non-U.S. Gov't
Summary
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The research examines the effect of centrifugation on Equus caballus sperm, with a focus on motility, viability and acrosomal integrity. The findings suggest that, while high centrifugal forces adversely impact the sperm, recovery rates may be enhanced.
Objective and Hypothesis
- The study aimed to investigate the influence of centrifugation on different aspects of the sperm of Equus caballus, a specific breed of horse, such as total and progressive motility, viability, and acrosomal integrity.
- The researchers hypothesized that even though high centrifugation forces would have a detrimental effect on the sperm, the chances of sperm recovery would be increased.
Methodology
- The researchers collected ejaculates from six stallions and thinned the concentration to 25×10(6) cells/mL.
- The samples were then subjected to different levels of centrifugation for a span of 10 minutes. The levels were (1) no centrifugation (NC), (2) centrifugation at 400 times gravity (400xg), (3) centrifugation at 900xg, or (4) centrifugation at 4500xg.
- The effects on sperm motility were assessed before and after centrifugation (referred to as Day 0), as well as after a 24-hour cooling period (referred to as Day 1).
- Samples were stained with SYBR-14/propidium iodide (PI) for viability and with PI/fluorescein isothiocyanate (FITC)-Peanut aglutinin (PNA) for acrosomal integrity assessment.
Findings
- Compared to other treatments, centrifugation at 4500xg reduced all end points measured (including motility, viability and acrosomal integrity).
- Lower centrifugation forces (400xg and 900xg) resulted in lower recovery rates compared to that of 4500xg.
- The 4500xg centrifugal force demonstrated a recovery rate of 97.9+/-2.8%, indicating minimal variations and consistency.
- Surprisingly, centrifugation at lower forces such as 400xg or 900xg did not cause any discernable damage to the equine sperm.
Conclusions
- The study concludes that further research is needed to pinpoint the optimal centrifugal force that achieves the highest recovery rate, least sperm damage, and does not negatively affect fertility.
- Specifically, further studies of centrifugal forces between 900xg and 4500xg are recommended.
Cite This Article
APA
Len JA, Jenkins JA, Eilts BE, Paccamonti DL, Lyle SK, Hosgood G.
(2009).
Immediate and delayed (after cooling) effects of centrifugation on equine sperm.
Theriogenology, 73(2), 225-231.
https://doi.org/10.1016/j.theriogenology.2009.09.003 Publication
Researcher Affiliations
- Department of Veterinary Clinical Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, Louisiana, USA. jlen@vetmed.lsu.edu
MeSH Terms
- Animals
- Centrifugation / veterinary
- Horses
- Male
- Sperm Motility
- Spermatozoa / physiology
- Tissue and Organ Harvesting / veterinary
Citations
This article has been cited 4 times.- Sugai N, Werre S, Cecere J, Balogh O. Defining an Optimal Range of Centrifugation Parameters for Canine Semen Processing. Animals (Basel) 2023 Apr 21;13(8).
- Neila-Montero M, Riesco MF, Alvarez M, Montes-Garrido R, Boixo JC, de Paz P, Anel-Lopez L, Anel L. Centrifugal force assessment in ram sperm: identifying species-specific impact. Acta Vet Scand 2021 Nov 4;63(1):42.
- Marzano G, Moscatelli N, Di Giacomo M, Martino NA, Lacalandra GM, Dell'Aquila ME, Maruccio G, Primiceri E, Chiriacò MS, Zara V, Ferramosca A. Centrifugation Force and Time Alter CASA Parameters and Oxidative Status of Cryopreserved Stallion Sperm. Biology (Basel) 2020 Jan 27;9(2).
- Sinagra L, Polisca A, Donato G, Caspanello T, Pettina G, Pastore S, De Majo M, Cristarella S, Quartuccio M, Zappone V. Enhancing canine semen quality through a second centrifugation after 48 hours of storage: a comparative study. Acta Vet Scand 2024 Sep 11;66(1):47.
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