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Home / Equine Research Bank / Anim Reprod / The interference of ozone gas in kinects and mitochondrial p...
Animal reproduction2021; 18(4); e20210075; doi: 10.1590/1984-3143-AR2021-0075

The interference of ozone gas in kinects and mitochondrial potential of equine sperm submitted on cryopreservation.

Abstract: The objective of this study was to evaluate the effects of the addition of different concentrations of ozone to quarter horse semen submitted to cryopreservation. Six ejaculates from four stallions were collected and were divided in four experimental groups: a control group (BotuCRIO® extender) and three other groups with BotuCRIO® ozonized at concentrations of 6, 8 and 12 μg of O/mL. The semen samples were diluted (200 x 10 spermatozoa/mL), filled in straws and frozen. After thawing (37 ºC, 30s), the samples were evaluated at 0, 30 and 60 minutes of incubation regarding sperm kinetics by a computer-assisted sperm analysis (CASA), and plasma membrane integrity (PMI), acrosome integrity (ACi) and mitochondrial membrane potential (MMP) by fluorescent probes. There was a reduction in the kinetic parameters total motility (TM), progressive motility (PM), curvilinear velocity (VCL), straight line velocity (VSL) and average path velocity (VAP) in all groups during the thermoresistance test (TT), a pattern also found in PMI and MMP analyses (p0.05) between the control and treatment (6, 8, and 12 μg of O/mL) groups, in any of the evaluated times for the kinetic parameters TM, linearity (LIN), straightness (STR), wobble index (WOB), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF). Regarding the VCL, VSL and VAP parameters, the group treated with 6 μg did not differ from the control or from 8 μg, but was higher than 12 μg at 30 and 60 minutes. ACi and PMI did not differ between groups (p>0.05), but PMI was lower in groups 8 μg and 12 μg compared to the control and 6 μg (p<0.05). It was concluded that the addition of ozone does not present beneficial effects for cryopreservation of equine semen at the concentrations used and decreases important parameters of fertility.
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Publication Date: 2021-12-17 PubMed ID: 34956402PubMed Central: PMC8697724DOI: 10.1590/1984-3143-AR2021-0075Google 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 investigates the influence of various concentrations of ozone gas on the preservation process of horse semen, particularly its impact on the sperm’s mobility and mitochondrial potential.

Outline of the Research Methodology

  • The research team collected six semen samples from four different stallions.
  • These samples were then divided into four groups. One operated as a control group with the semen frozen in a BotuCRIO® extender. The other three groups used the same extender, but it had been ozonized at either 6, 8, or 12 μg of O/mL concentrations.
  • After diluting the semen samples to a ratio of 200 x 10 spermatozoa/mL, the researchers filled these into straws and then froze them.
  • Once the samples were thawed (at 37 ºC, for 30 seconds), they were evaluated at 0, 30, and 60-minute marks. The parameters investigated were sperm kinetics, plasma membrane integrity (PMI), acrosome integrity (ACi), and mitochondrial membrane potential (MMP), through the aid of a computer-assisted sperm analysis (CASA) tool and various fluorescent probes.

Findings of the Research

  • The analysis revealed a drop in the total motility (TM), progressive motility (PM), curvilinear velocity (VCL), straight line velocity (VSL), and average path velocity (VAP) of all the groups, including the control group, after the thermoresistance test (TT). Similar drops were observed in PMI and MMP analytics (p<0.05).
  • No significant difference (p>0.05) was observed between the control group and the ozonized group in terms of kinetic parameters such as TM, linearity (LIN), straightness (STR), wobble index (WOB), amplitude of lateral head displacement (ALH), and beat cross frequency (BCF).
  • However, certain parameters including the VCL, VSL, and VAP values of the group treated with 6 μg ozone did not show a significant difference from control or 8 μg groups but recorded higher values than the 12 μg group at the 30 and 60-minute stages of observation.
  • No significant difference was observed in the acrosome integrity (ACi) across all groups. Nevertheless, the PMI was lower in the 8 μg and 12 μg ozone groups compared to the control group and the 6 μg group (p<0.05).

Conclusion of the Research

  • The researchers concluded that the addition of ozone gas during the cryopreservation process does not necessarily yield beneficial effects, given the studied concentrations. Moreover, the use of ozone gas impacted sperm viability and led to a decrease in crucial fertility parameters.

Cite This Article

APA
Macêdo IN, Arruda LCP, de Santana BB, de Moura TCM, Guerra MMP, Bezerra DG, Carneiro GF, Silva SV. (2021). The interference of ozone gas in kinects and mitochondrial potential of equine sperm submitted on cryopreservation. Anim Reprod, 18(4), e20210075. https://doi.org/10.1590/1984-3143-AR2021-0075

Publication

Animal reproduction
ISSN: 1984-3143
NlmUniqueID: 101272804
Country: Brazil
Language: English
Volume: 18
Issue: 4
Pages: e20210075
PII: e20210075

Researcher Affiliations

Macêdo, Iara Nóbrega
  • Programa de Pós-Graduação em Ciência Animal, Centro de Ciências Agrárias, Universidade Federal da Paraíba, Areia, PB, Brasil.
Arruda, Lucia Cristina Pereira
  • Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco, Recife, PE, Brasil.
de Santana, Breno Barros
  • Central Monte Verde de Reprodução Equina, Sairé, PE, Brasil.
de Moura, Thalles Cloves Maciel
  • Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco, Recife, PE, Brasil.
Guerra, Maria Madalena Pessoa
  • Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco, Recife, PE, Brasil.
Bezerra, Diogo Gutemberg
  • Central Monte Verde de Reprodução Equina, Sairé, PE, Brasil.
Carneiro, Gustavo Ferrer
  • Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco, Recife, PE, Brasil.
Silva, Sildivane Valcácia
  • Programa de Pós-Graduação em Ciência Animal, Centro de Ciências Agrárias, Universidade Federal da Paraíba, Areia, PB, Brasil.
  • Centro de Biotecnologia, Universidade Federal da Paraíba, João Pessoa, PB, Brasil.

Conflict of Interest Statement

Conflict of interest: The authors have no conflict of interest to declare.

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Citations

This article has been cited 1 times.
  1. Fanelli D, Moroni R, Sala G, Melanie P, Tarabella I, Telleschi N, Maltinti S, Giorgi M, Barsotti G, Passamonti F, Marmorini P, Rota A, Camillo F, Panzani D. Impact of low-dose ozone supplementation on motility parameters and bacterial growth in horse cryopreserved semen. Vet Anim Sci 2024 Sep;25:100380.
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