A spectrophotometric procedure for the determination of objective measurements of equine spermatozoan motility.
Abstract: A spectrophotometric procedure was developed and evaluated for the objective measurement of equine spermatozoan motility. A 100 mul sample of a sperm suspension, prepared by the removal of seminal plasma, was layered under a column of optically clear medium in a specially designed spectrophotometric cuvette maintained at 37 degrees C. Changes in light transmittance above the interface of the sperm suspension and medium were recorded on chart paper. As sperm cells swam into the medium, a decrease in light transmittance was recorded as a deflection on the chart paper. Chart recordings were analyzed for the height (cm) and time (min) to the peak deflection. To standardize the procedure, a fixed number of cells (1x10(9)) were used to prepare suspensions of 300x10(6) cells/ml. Coefficients of variation for mean values obtained under these conditions after the evaluation of five ejaculates from a given stallion were estimated at between 10 and 12%. Correlations between swim-up measurements and computer-assisted semen analysis demonstrated that the percentage of motile cells and mean velocity (mum/sec) of motile cells influenced swim-up measurements. Described here is a simple and inexpensive procedure to determine objective measurements of spermatozoan motility that may have application in semen evaluation and fertility testing in the stallion.
Publication Date: 1989-05-01 PubMed ID: 16726611DOI: 10.1016/0093-691x(89)90477-9Google Scholar: Lookup
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Summary
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This research outlines the development and assessment of a spectrophotometric procedure for objectively measuring horse sperm movement. The method shows promise for evaluating semen and testing fertility in stallions.
Developing the Procedure
- The researchers devised a spectrophotometric procedure to objectively measure the mobility of horse sperm cells or spermatozoa. This process involves using light to measure the amount of a specific substance in a solution.
- In their experiment, a suspension of sperm, from which seminal plasma had been removed, was layered under a clear medium in a spectrophotometric cuvette, a container designed to hold samples for spectroscopic analysis. The sample used was 100 microliters (µL) while the cuvette was kept at a temperature of 37 degrees Celsius, similar to body temperature.
Recording and Analyzing Data
- Changes in the light transmission above the point of intersection between the sperm suspension and the medium were recorded visually on chart paper.
- As the sperm cells swam up into the medium, there was a corresponding decrease in light transmittance, which was noted as a deflection on the chart paper.
- Recordings on the chart were then analyzed to determine the height (in centimeters) and time (in minutes) to the peak deflection.
Procedure Standardization
- For the sake of standardization, the researchers used a fixed number of cells (1×10^9) to create suspensions with a concentration of 300×10^6 cells per milliliter.
- By repeating the measurement process on five different ejaculates from the same stallion, the average values obtained demonstrated a coefficient of variation estimated between 10 and 12%. This measure refers to a statistical calculation that helps to understand the degree of variation from the average.
Correlations and Applications
- Upon comparing the results with computer-assisted semen analysis, it was found that both the percentage of moving cells and the average speed of those cells influenced the swim-up measurements.
- In conclusion, this research provides a straightforward and cost-efficient procedure for obtaining objective measurements of sperm mobility. This could be useful in evaluating stallion semen and assessing their fertility.
Cite This Article
APA
Jasko DJ, Smith K, Little TV, Lein D, Foote RH.
(1989).
A spectrophotometric procedure for the determination of objective measurements of equine spermatozoan motility.
Theriogenology, 31(5), 945-954.
https://doi.org/10.1016/0093-691x(89)90477-9 Publication
Researcher Affiliations
- New York State College of Veterinary Medicine Cornell University, Ithaca, NY 14853 USA.
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