Lactoferrin increases sperm membrane functionality of frozen equine semen.
Abstract: During cryopreservation, sperm was submitted to an increase in reactive oxygen species generation. This work aimed to improve the quality of frozen equine sperm after the addition of antioxidants lactoferrin (Lf) and catalase (Cat) to a freezing extender. Semen from six stallions was frozen with the extenders: F1) control, INRA 82 freezing extender, F2) F1 + 500 μg/ml Lf and F3) F1 + 200 IU/ml Cat. After thawing, sperm motility parameters, membrane functionality and integrity, and acrosome integrity and spontaneous acrosome-reacted sperm were evaluated with a computer-assisted sperm analysis, a hypoosmotic swelling test and epifluorescent microscopy, respectively. Nitrite, hydroperoxide and iron concentrations of frozen semen were measured with spectrophotometry. The percentage of functional membrane sperm treated with Lf was higher (50.7% ± 11.6%) compared to that of the control (37.6% ± 15.6%), while the iron (61.4 ± 11.6 vs 73.3 ± 13.8 mg/dl) and nitrite concentrations (16.3 ± 7.1 vs 25.9 ± 4.2 μM/μg protein) were lower, respectively (p < .05). Thus, it can be suggested that Lf protect stallion spermatozoon during freezing as it has increased the percentage of sperm with functional membrane and decreased the lipid oxidant agents.
© 2018 Blackwell Verlag GmbH.
Publication Date: 2018-02-12 PubMed ID: 29431233DOI: 10.1111/rda.13148Google Scholar: Lookup
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- Journal Article
Summary
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The research investigates the effectiveness of using antioxidants lactoferrin (Lf) and catalase (Cat) to improve the quality of frozen horse semen. Significant findings reveal that lactoferrin enhances the percentage of functional sperm membrane and reduces lipid oxidant agents.
Study Purpose and Methodology
- The aim of this research was to improve the quality of frozen horse sperm by adding the antioxidants lactoferrin (Lf) and catalase (Cat) to a freezing compound. These antioxidants were utilized with the intention of managing the increase in reactive oxygen species (ROS) that usually occurs during cryopreservation. Reactive oxygen species are chemically reactive molecules containing oxygen, which can cause significant damage to cell structures.
- The semen from six male horses was frozen with different extenders: one, a control group using the INRA 82 freezing extender; the second group was given the same freezing extender but with an addition of 500 μg/ml of Lf, while the third group had 200 IU/ml Cat added into the extender.
Experimental Testing and Evaluation
- After thawing, sperms’ motility parameters, membrane functionality and integrity, and acrosome integrity and spontaneous acrosome-reacted sperm were evaluated. Techniques used for the analysis included computer-assisted sperm analysis, a hypoosmotic swelling test and epifluorescent microscopy respectively.
- The researchers used spectrophotometry to measure the concentrations of nitrite, hydroperoxide, and iron from the frozen semen.
Results and Significance
- The results indicated that the percentage of functional membrane sperm treated with lactoferrin was significantly higher at 50.7% compared to the control group which was at 37.6%. Furthermore, the lactoferrin treatment resulted in lower iron and nitrite concentrations.
- In conclusion, these results reveal that lactoferrin can protect horse sperm during freezing. It accomplishes this by enhancing the percentage of sperm with functional membrane and lowering the presence of lipid oxidant agents. This is a significant insight for the scientific and veterinary community as it can impact methods of preserving and enhancing the quality of animal semen.
Cite This Article
APA
Martins HS, da Silva GC, Cortes SF, Paes FO, Martins Filho OA, Araujo M, Stahlberg R, Lagares MA.
(2018).
Lactoferrin increases sperm membrane functionality of frozen equine semen.
Reprod Domest Anim, 53(3), 617-623.
https://doi.org/10.1111/rda.13148 Publication
Researcher Affiliations
- Departamento de Clinica e Cirurgia Veterinárias da Escola de Veterinária da, Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, Brazil.
- Departamento de Farmacologia do Instituto de Ciências Biológicas da UFMG, Belo Horizonte, Brazil.
- Departamento de Farmacologia do Instituto de Ciências Biológicas da UFMG, Belo Horizonte, Brazil.
- Departamento de Clinica e Cirurgia Veterinárias da Escola de Veterinária da, Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, Brazil.
- Centro de Pesquisas René Rachou - Fiocruz, Laboratório de Biomarcadores de Diagnóstico e Monitoração, Belo Horizonte, Brazil.
- Centro de Pesquisas René Rachou - Fiocruz, Laboratório de Biomarcadores de Diagnóstico e Monitoração, Belo Horizonte, Brazil.
- Faculdade de Medicina Veterinária da Pontifícia, Universidade Católica- PUC Minas, Betim, Brazil.
- Departamento de Clinica e Cirurgia Veterinárias da Escola de Veterinária da, Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, Brazil.
MeSH Terms
- Acrosome Reaction
- Animals
- Antioxidants
- Catalase / pharmacology
- Cell Membrane / physiology
- Cryopreservation / methods
- Cryopreservation / veterinary
- Cryoprotective Agents / pharmacology
- Horses
- Lactoferrin / pharmacology
- Male
- Semen Analysis / veterinary
- Semen Preservation / veterinary
- Sperm Motility
- Spermatozoa / cytology
- Spermatozoa / drug effects
Citations
This article has been cited 9 times.- Ghaly MF, Abougabal KM, Allam AA, Palani A, Mostafa T. Assessment of Seminal Lactoferrin Levels in Oligoasthenoterato-zoospermic Men with Varicocele. J Reprod Infertil 2025 Apr-Jun;26(2):119-125.
- Celedon JJ, Corder ML, McConnell M, Songsasen N, Nagashima JB. Proteome of seminal plasma and sperm associated with sperm survival following cryopreservation in the Red Wolf (Canis rufus). Sci Rep 2025 Oct 29;15(1):37882.
- Mahran AM, Abd-Almalak MS, Helbawi FM, Mekkawy MM. Detection of Lactoferrin and Iron in Seminal Plasma and Their Possible Relation to Semen Parameters and Infertility in Varicocele: A Cross-sectional Study. J Hum Reprod Sci 2024 Oct-Dec;17(4):284-291.
- Hungerford AJ, Bakos HW, Aitken RJ. Analysis of sperm separation protocols for isolating cryopreserved human spermatozoa. Reprod Fertil 2023 Apr 1;4(2).
- Cao X, Ren Y, Lu Q, Wang K, Wu Y, Wang Y, Zhang Y, Cui XS, Yang Z, Chen Z. Lactoferrin: A glycoprotein that plays an active role in human health. Front Nutr 2022;9:1018336.
- Su J, Wang C, Song Y, Yang Y, Cao G. Effect of lactoferrin on ram sperm motility after cryopreservation. Anim Biosci 2022 Sep;35(9):1351-1359.
- Fuentes-Albero MC, González-Brusi L, Cots P, Luongo C, Abril-Sánchez S, Ros-Santaella JL, Pintus E, Ruiz-Díaz S, Barros-García C, Sánchez-Calabuig MJ, García-Párraga D, Avilés M, Izquierdo Rico MJ, García-Vázquez FA. Protein Identification of Spermatozoa and Seminal Plasma in Bottlenose Dolphin (Tursiops truncatus). Front Cell Dev Biol 2021;9:673961.
- Vitorino Carvalho A, Soler L, Thélie A, Grasseau I, Cordeiro L, Tomas D, Teixeira-Gomes AP, Labas V, Blesblois E. Proteomic Changes Associated With Sperm Fertilizing Ability in Meat-Type Roosters. Front Cell Dev Biol 2021;9:655866.
- Len JS, Koh WSD, Tan SX. The roles of reactive oxygen species and antioxidants in cryopreservation. Biosci Rep 2019 Aug 30;39(8).
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