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Home / Equine Research Bank / Animals (Basel) / Single Layer Centrifugation Improves the Quality of Fresh Do...
Animals : an open access journal from MDPI2020; 10(11); 2128; doi: 10.3390/ani10112128

Single Layer Centrifugation Improves the Quality of Fresh Donkey Semen and Modifies the Sperm Ability to Interact with Polymorphonuclear Neutrophils.

Abstract: This study sought to determine whether single layer centrifugation (SLC) of fresh donkey semen with Equicoll has any impact on sperm quality parameters and on the modulation of endometrial reaction following semen deposition using an in vitro model. Seventeen ejaculates from five jackasses were obtained using an artificial vagina and diluted in a skim-milk extender. Samples were either selected through SLC (Equicoll) or non-treated (control). Two experiments were performed. The first one consisted of incubating selected or non-selected spermatozoa at 38 °C for 180 min. Integrity and lipid disorder of sperm plasma membrane, mitochondrial membrane potential, and intracellular levels of calcium and reactive oxygen species were evaluated at 0, 60, 120, and 180 min. In the second experiment, polymorphonuclear neutrophils (PMN) isolated from jennies blood were mixed with selected and unselected spermatozoa. Interaction between spermatozoa and PMN was evaluated after 0, 60, 120, and 180 min of co-incubation at 38 °C. SLC-selection increased the proportions of spermatozoa with an intact plasma membrane and low lipid disorder, of spermatozoa with high mitochondrial membrane potential and with high calcium levels, and of progressively motile spermatozoa. In addition, selection through SLC augmented the proportion of phagocytosed spermatozoa, which supported the modulating role of seminal plasma proteins on sperm-PMN interaction. In conclusion, SLC of fresh donkey semen increases the proportions of functionally intact and motile spermatozoa, and appears to remove the seminal plasma proteins that inhibit sperm-PMN binding.
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Publication Date: 2020-11-16 PubMed ID: 33207812PubMed Central: PMC7696916DOI: 10.3390/ani10112128Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research paper delves into the examination of the effects of single layer centrifugation (SLC) of fresh donkey semen on sperm quality and the ability of the sperm to interact with a type of white blood cell. The results indicate that the centrifugation method significantly enhances sperm quality and their interactions with white blood cells.

Methods and Procedures

  • The researchers used semen from five donkeys, collected with an artificial vagina, and diluted in a milk-based substance.
  • The semen was either put through the SLC procedure with a substance known as Equicoll or left untreated as a control group.
  • Two separate experiments were carried out. The first involved incubating the semen at a specific temperature for a period of 180 minutes. Various aspects of the sperm cells, such as membrane integrity, lipid order, mitochondrial stability, and levels of calcium and reactive oxygen species, were observed at different intervals.
  • The second experiment involved the extraction of a specific type of white blood cell, polymorphonuclear neutrophils (PMN), from female donkey blood and introducing it to the selected and unselected sperm. The interaction was then reviewed at given time points.

Results of the Research

  • The results showed that the SLC process improved the semen sample overall. It increased the number of sperm cells that had an intact plasma membrane and low lipid disorder, which indicates healthy and functional sperm cells.
  • The processed semen also yielded higher counts of sperm cells with good mitochondrial membrane potential, high calcium levels, and progressive motility. This suggests a better fertility potential in the sperm cells.
  • Moreover, the SLC method also increased the proportion of sperm cells that were effectively absorbed by the white blood cells or ‘phagocytosed’, implying a key role of seminal plasma proteins in regulating the interaction between sperm cells and white blood cells.

Conclusions Drawn

  • The study concluded that the SLC centrifugation of fresh donkey semen effectively enhances the quality and functionality of the sperm cells.
  • This process also appears to remove the seminal plasma proteins that usually inhibit sperm and white blood cell interactions, hence facilitating the process of sperm absorption by these cells.

Overall, this paper researches an effective mechanism to enhance the quality of donkey semen by employing a single layer centrifugation technique. The enhanced sperm cells show better functionality and interaction with female white blood cells. This could potentially be promising for further study concerning fertility issues in the reproductive field of animal husbandry and beyond.

Cite This Article

APA
Papas M, Catalán J, Recuero S, Morrell JM, Yeste M, Miró J. (2020). Single Layer Centrifugation Improves the Quality of Fresh Donkey Semen and Modifies the Sperm Ability to Interact with Polymorphonuclear Neutrophils. Animals (Basel), 10(11), 2128. https://doi.org/10.3390/ani10112128

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 10
Issue: 11
PII: 2128

Researcher Affiliations

Papas, Marion
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, E-08193 Bellaterra (Cerdanyola del Vallès), Spain.
Catalán, Jaime
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, E-08193 Bellaterra (Cerdanyola del Vallès), Spain.
Recuero, Sandra
  • Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, E-17003 Girona, Spain.
  • Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, E-17003 Girona, Spain.
Morrell, Jane M
  • Clinical Sciences, Swedish University of Agricultural Sciences, Box 7054, SE-75007 Uppsala, Sweden.
Yeste, Marc
  • Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, E-17003 Girona, Spain.
  • Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, E-17003 Girona, Spain.
Miró, Jordi
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, E-08193 Bellaterra (Cerdanyola del Vallès), Spain.

Conflict of Interest Statement

J.M.M. is the inventor and one of the patent holders of the colloid formulation used in this study. The other authors declare no conflicts of interest.

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Citations

This article has been cited 2 times.
  1. Morrell JM, Rocha A. A Novel Approach to Minimising Acute Equine Endometritis That May Help to Prevent the Development of the Chronic State.. Front Vet Sci 2021;8:799619.
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  2. Yánez-Ortiz I, Catalán J, Delgado-Bermúdez A, Carluccio A, Miró J, Yeste M. Addition of Reduced Glutathione (GSH) to Freezing Medium Reduces Intracellular ROS Levels in Donkey Sperm.. Vet Sci 2021 Dec 2;8(12).
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