Sperm transport and endometrial inflammatory response in mares after artificial insemination with cryopreserved spermatozoa.
Abstract: This study aimed to determine whether the insemination site and dose with cryopreserved sperm of reproductively normal mares affect the sperm population in uterine tubes and the intensity of endometrial inflammatory response. Experimental subjects were estrous mares inseminated, in the mid-uterine body (Body) or the tip of the uterine horn (Tip), ipsilateral to the dominant follicle, with one 0.5 mL straw with 50 × 106 sperm (50) or with eight straws with 50 × 106 sperm/straw (400). Mares were slaughtered 2 h, 4 h and 12 h after artificial insemination (AI) and randomly assigned to following groups: Body 50 (n = 19) (2 h, 4 h or 12 h); Tip 50 (n = 29) (2 h, 4 h, or 12 h); Body 400 (n = 24) (2 h, 4 h, or 12 h); Tip 400 (n = 21) (2 h, 4 h, or 12 h). A Control group (n = 16) was not inseminated. After slaughter, uterine tubes were separated from uterus, and uteri and tubes flushed with phosphate-buffered saline (PBS). After flushing, an endometrial sample was collected from ipsilateral and contralateral horns and mid-uterus body for further histopathological examination. A sample of each uterine tube flushing was examined for sperm count, and a sample of each uterine flushing was used for polymorphonuclear neutrophils (PMNs) count. Data were analyzed using PROC GLM from SASv9.4. Insemination time, site, sperm dose, and their interactions were considered independent variables and sperm and PMNs numbers dependent variables. Deep horn insemination increased ipsilateral uterine tube sperm number without an increase in the inflammatory reaction compared with the uterine body insemination. The higher the insemination dose, the higher the uterine tubes' sperm number and inflammatory reaction, with a quicker resolution. In conclusion, the insemination site and dose affected sperm in the uterine tubes, while post-insemination time and dose influenced the inflammatory reaction.
Copyright © 2020 Elsevier Inc. All rights reserved.
Publication Date: 2020-09-15 PubMed ID: 32961353DOI: 10.1016/j.theriogenology.2020.09.021Google Scholar: Lookup
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- Journal Article
- Randomized Controlled Trial
- Veterinary
Summary
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The study investigates how different factors of artificial insemination with frozen sperm in healthy mares affect both the number of sperm in the uterine tubes and the intensity of the immune response in the uterine lining.
Objective and Methods
- The research aims to comprehend whether the site of insemination and the dose of cryopreserved sperm in healthy female horses impact the sperm population in uterine tubes and the intensity of the endometrial inflammatory response.
- The mares in estrus were artificially inseminated either at the middle part of the uterus (referred to as the body) or at the tip of the uterine horn, on the side where the dominant follicle was located.
- The mares were given either a small dosage (one 0.5 mL straw containing 50 x 106 sperm) or a large dosage (eight straws each containing 50 x 106 sperm/straw) of sperm.
- After the inseminations, mares were slaughtered at intervals of 2 hours, 4 hours, and 12 hours. Each mare was randomly placed into one of four treatment groups: Body 50, Tip 50, Body 400, or Tip 400. A control group of mares was not inseminated.
Procedure
- Following slaughter, the uterine tubes were separated from the uterus, and both uterus and tubes were flushed with phosphate-buffered saline (PBS).
- An endometrial tissue sample was taken from the side of the uterus connected to the dominant follicle, the opposite side, and the central region to undergo histopathological analysis.
- Flush samples from each uterine tube were evaluated for sperm count, whereas flush samples from the uterus were used to count polymorphonuclear neutrophils (PMNs), a type of white blood cell integral to the immune response.
Data Analysis
- Data was examined using PROC GLM in the statistical software SASv9.4. The independent variables were insemination time, insemination site, and sperm dose, while the dependent variables were sperm count and PMN count.
Findings
- Deep horn insemination led to increased sperm count in the uterine tube on the side of the dominant follicle, but no corresponding increase in inflammatory response, compared to insemination in the body of the uterus.
- Higher doses of sperm led to more sperm in the uterine tubes and a stronger inflammatory response, which resolved more rapidly.
Conclusion
- Overall, the insemination site and dose did impact the number of sperm in the uterine tubes. The inflammation after insemination was influenced by both the time post-insemination and the sperm dose.
Cite This Article
APA
Cazales N, Estradé MJ, Pereyra F, Fiala-Rechsteiner SM, Mattos RC.
(2020).
Sperm transport and endometrial inflammatory response in mares after artificial insemination with cryopreserved spermatozoa.
Theriogenology, 158, 180-187.
https://doi.org/10.1016/j.theriogenology.2020.09.021 Publication
Researcher Affiliations
- Departamento de Reproducción Animal, Facultad de Veterinaria, UDELAR, Montevideo, Uruguay; Reprolab, Faculdade de Veterinária, UFRGS, Porto Alegre, RS, Brazil; Bolsista CAPES/UDELAR, Brazil.
- Departamento de Reproducción Animal, Facultad de Veterinaria, UDELAR, Montevideo, Uruguay.
- Departamento de Reproducción Animal, Facultad de Veterinaria, UDELAR, Montevideo, Uruguay.
- Historep, Faculdade de Veterinária, UFPel, Pelotas, RS, Brazil.
- Reprolab, Faculdade de Veterinária, UFRGS, Porto Alegre, RS, Brazil. Electronic address: rcmattos@ufrgs.br.
MeSH Terms
- Animals
- Cryopreservation / veterinary
- Female
- Horses
- Insemination, Artificial / veterinary
- Male
- Sperm Count / veterinary
- Sperm Transport
- Spermatozoa
- Uterus
Conflict of Interest Statement
Declaration of competing interest The authors declare that they have no conflicts of interest with the contents of this article.
Citations
This article has been cited 2 times.- Zhao Y, Liu Y, Tao J, Cao J, Lin Y, He Q, Fang X, Yun S, Du M, Su S, Bao T, Bai D, Zhang X, Dugarjaviin M. Isolation, identification and comparative genomic analysis of Lactobacillus salivarius from Mongolian horse vagina. Front Microbiol 2025;16:1635639.
- Hidalgo AI, Ulloa-Leal C, Gajardo G, López G, Carretta D, Burgos RA, Ratto M. Ovulation Induced by Intrauterine Seminal Plasma Increases Total Protein, PGE2, IL-8, and IL-1β in Uterine Fluid of Llamas (Lama glama). Animals (Basel) 2023 Feb 5;13(4).
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