Effects of staining method and clinician experience on the evaluation of stallion sperm morphology.
Abstract: Evaluation of sperm morphology is part of the assessment of fertility in human and animal reproduction. Analyses can be performed using different techniques, including the use of staining methods In our prospective study, the morphology of equine sperm was evaluated using 3 staining methods Diff-Quik, eosin-nigrosin, and SpermBlue, the latter being a relatively new stain commonly used for human sperm. Our hypotheses were that (1) SpermBlue allows easier assessment of horse sperm morphology and facilitates better identification of sperm abnormalities, and (2) sperm morphology classification differs depending on the evaluator's experience. Semen was obtained from various horse breeds; 40 samples from stallions between 2 and 15 years of age were collected during the 2016 breeding season and stored in a 2% buffered formaldehyde solution until processing. For each sample, 3 semen smears were made and stained with Diff-Quik, eosin-nigrosin, and SpermBlue. All morphological parameters were then evaluated blindly using a light microscope by a novice evaluator and a more experienced evaluator. For each slide, 200 spermatozoa were examined randomly and classified according to their characteristics. For the identification of morphologically normal spermatozoa, no significant difference between evaluators was found with any of the staining methods used. In contrast, significant differences between evaluators were observed in the classification of some anomalies affecting mainly the midpiece and the tail. Poor dye fixation was observed with SpermBlue.
Copyright © 2017 Elsevier B.V. All rights reserved.
Publication Date: 2017-11-29 PubMed ID: 29198956DOI: 10.1016/j.anireprosci.2017.11.021Google Scholar: Lookup
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- Journal Article
Summary
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The research paper discusses a study on the evaluation of horse sperm morphology using three staining methods and examining how the evaluator’s experience influences the classification of the sperm morphology. The key findings include the influence of experience on detection of anomalies and the performance of the staining methods used.
Overview of the Study
- The study took place over the course of the 2016 breeding season and involved examining the sperm of stallions aged between 2 and 15 years.
- Three staining methods were used – Diff-Quik, eosin-nigrosin, and SpermBlue. Each sample was tested with all three stains.
- Two evaluators analysed the sample, one novice and one experienced, and all evaluations were done blindly.
Staining Methods
- The purpose of the different staining methods is to help identify and differentiate the various components of the sperm.
- Diff-Quik and eosin-nigrosin are established methods while SpermBlue is a new method typically used for human sperm.
- However, the dye fixation with SpermBlue was noted to be poor in this study.
Evaluator’s Experience and Sperm Morphology Classification
- Each evaluator classified 200 randomly selected spermatozoa, based on their characteristics.
- No significant difference between the evaluations of the novice and experienced evaluators was found when identifying morphologically normal spermatozoa.
- However, significant differences between the evaluators were observed when classifying certain anomalies, particularly in the midpiece and the tail of the sperm. This suggests that experience level does affect the detection and classification of certain sperm anomalies.
Hypotheses Confirmed or Rejected
- The hypothesis that SpermBlue allows for easier assessment of horse sperm morphology was not fully confirmed due to its poor dye fixation.
- However, the hypothesis that sperm morphology classification varies depending on the evaluator’s experience was confirmed. A higher level of experience helped in identifying certain types of anomalies more accurately.
Cite This Article
APA
Murcia-Robayo RY, Jouanisson E, Beauchamp G, Diaw M.
(2017).
Effects of staining method and clinician experience on the evaluation of stallion sperm morphology.
Anim Reprod Sci, 188, 165-169.
https://doi.org/10.1016/j.anireprosci.2017.11.021 Publication
Researcher Affiliations
- Department of Clinical Sciences, Faculty of Veterinary Medicine, Université de Montréal, Saint-Hyacinthe, Q, Canada.
- Department of Clinical Sciences, Faculty of Veterinary Medicine, Université de Montréal, Saint-Hyacinthe, Q, Canada.
- Faculty of Veterinary Medicine, Université de Montréal, Saint-Hyacinthe, Q, Canada.
- Department of Clinical Sciences, Faculty of Veterinary Medicine, Université de Montréal, Saint-Hyacinthe, Q, Canada. Electronic address: mouhamadou.diaw@umontreal.ca.
MeSH Terms
- Animals
- Horses / physiology
- Male
- Observer Variation
- Semen Analysis / methods
- Semen Analysis / veterinary
- Spermatozoa / physiology
- Staining and Labeling
Citations
This article has been cited 6 times.- Baqir Al-Dhalimy AM, Alabsawy SK, Al-Mousaw M, Al-Dhalemi DM. Ameliorated Effect of Ascorbic Acid and Selenium against the Stress Effect on Sperm Quality of Rats.. Arch Razi Inst 2021 Oct;76(4):1137-1142.
- Ebel F, Ulloa O, Strobel P, Ramírez-Reveco A. Semen Quality and Freezability Analyses in the Ejaculates of Two Poitou Donkeys in the Southern Hemisphere.. Front Vet Sci 2021;8:662887.
- Bustani GS, Baiee FH. Semen extenders: An evaluative overview of preservative mechanisms of semen and semen extenders.. Vet World 2021 May;14(5):1220-1233.
- Alamaary MS, Haron AW, Hiew MWH, Ali M. Effects of cysteine and ascorbic acid in freezing extender on sperm characteristics and level of enzymes in post-thawed stallion semen.. Vet Med Sci 2020 Nov;6(4):666-672.
- Baily MP, Avila F, Das PJ, Kutzler MA, Raudsepp T. An Autosomal Translocation 73,XY,t(12;20)(q11;q11) in an Infertile Male Llama (Lama glama) With Teratozoospermia.. Front Genet 2019;10:344.
- Alamaary MS, Haron AW, Ali M, Hiew MWH, Adamu L, Peter ID. Effects of four extenders on the quality of frozen semen in Arabian stallions.. Vet World 2019 Jan;12(1):34-40.
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