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Home / Equine Research Bank / Theriogenology / Comparison of an extender containing defined milk protein fr...
Theriogenology2006; 66(5); 1115-1122; doi: 10.1016/j.theriogenology.2006.03.006

Comparison of an extender containing defined milk protein fractions with a skim milk-based extender for storage of equine semen at 5 degrees C.

Abstract: A problem of semen extenders based on milk or egg yolk is the fact that these biological products consist of a variety of substances. Extenders containing only components with clearly protective effects on spermatozoa would thus be an advantage. In this study, we have compared the effects of an extender containing defined caseinates and whey proteins only (EquiPro, defined milk protein extender) with skim milk extender on equine spermatozoa during cooled storage. The defined milk protein extender was used with and without the antioxidant N-acetyl cysteine (NAC). In a second experiment, semen was diluted with PBS or defined milk protein extender and was either stored directly or 90% of seminal plasma was removed by centrifugation and replaced by defined milk protein extender before storage. In both experiments, eight stallions were available for semen collections. Motility, velocity and membrane integrity of spermatozoa were determined by CASA immediately after semen processing and after 24, 48 and 72 h of storage at 5 degrees C. Total motility after 24 h of storage was lowest in semen diluted with PBS (p<0.05 versus all extenders). At 48 and 72 h, motility of spermatozoa in defined milk protein extender was significantly (p<0.05) higher than in PBS or skim milk extender. Velocity of spermatozoa after storage was highest in defined milk protein extender. Membrane integrity after storage was significantly (p<0.05) lower in semen diluted with PBS than in semen diluted with both extenders. Addition of NAC was without effect on the examined parameters. Centrifugation further increased the percentage of motile and membrane-intact spermatozoa in the defined milk protein extender (p<0.05). Velocity of spermatozoa in this extender was not negatively affected by centrifugation.
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Publication Date: 2006-04-18 PubMed ID: 16620943DOI: 10.1016/j.theriogenology.2006.03.006Google Scholar: Lookup
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  • Comparative Study
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

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 examines the effectiveness of a semen extender containing only defined caseinates and whey proteins (EquiPro) in preserving equine spermatozoa during cooled storage, compared to a traditional skim milk-based extender.

Objective of Research

  • The study primarily focuses on assessing the viability of an extender, EquiPro, constituted solely of defined milk protein fractions, caseinates, and whey proteins, in storing equine semen at 5 degrees Celsius. This is done against the backdrop of traditional extenders composed of various substances invariably found in milk or egg yolk, with the view that an extender of specific components with clearly sperm-protective characteristics would be beneficial.

Research Design and Method

  • Two experiments were conducted with semen obtained from eight stallions. EquiPro was tested with and without an antioxidant, N-acetyl cysteine (NAC).
  • The first experiment involved diluting semen with either PBS (Phosphate-Buffered Saline) or EquiPro. In the second, 90% of the seminal plasma was removed via centrifugation and replaced with EquiPro.
  • Motility, velocity and membrane integrity of the spermatozoa were determined through Computer-Assisted Sperm Analysis (CASA) immediately after semen processing, as well as after 24, 48, and 72 hours of storage at 5 degrees Celsius.

Results of the Research

  • The findings revealed that total motility after 24 hours of storage was lowest for PBS-diluted semen.
  • For durations of 48 and 72 hours, semen stored in EquiPro exhibited higher motility rates than those in PBS or skim milk extender. The velocity of spermatozoa post-storage was also highest in the defined milk protein extender.
  • Comparatively, membrane integrity was significantly lower in semen diluted with PBS than those diluted with both extenders.
  • Contrary to what might be expected, the addition of NAC had no observable effect on any studied parameters. However, using centrifugation helped increase the percentage of motile and membrane-intact spermatozoa in the semen stored with EquiPro.

Implications of the Research

  • Overall, the results suggest that the purpose-specific extender, EquiPro, performed better than traditional extenders in preserving equine semen during cooled storage, thereby addressing a longstanding issue regarding the complex, multi-substance composition of traditional extenders.
  • Improvement was noted in sperm motility, velocity, and membrane integrity, particularly when centrifugation was applied. This highlights the potential of defined milk protein extenders to reliably and effectively maintain the viability of spermatozoa for artificial insemination in horses.

Cite This Article

APA
Pagl R, Aurich JE, Müller-Schlösser F, Kankofer M, Aurich C. (2006). Comparison of an extender containing defined milk protein fractions with a skim milk-based extender for storage of equine semen at 5 degrees C. Theriogenology, 66(5), 1115-1122. https://doi.org/10.1016/j.theriogenology.2006.03.006

Publication

Theriogenology
ISSN: 0093-691X
NlmUniqueID: 0421510
Country: United States
Language: English
Volume: 66
Issue: 5
Pages: 1115-1122

Researcher Affiliations

Pagl, Roland
  • Klinik für Geburtshilfe, Gynäkologie und Andrologie, Veterinärmedizinische Universität, 1210 Vienna, Austria. roland.pagl@vu-wien.ac.at
Aurich, Jörg E
    Müller-Schlösser, Frank
      Kankofer, Marta
        Aurich, Christine

          MeSH Terms

          • Animals
          • Cold Temperature
          • Cysteine / pharmacology
          • Horses / physiology
          • Image Processing, Computer-Assisted / methods
          • Male
          • Milk Proteins / pharmacology
          • Semen Preservation / methods
          • Semen Preservation / veterinary
          • Sperm Motility
          • Spermatozoa / physiology
          • Temperature
          • Time Factors

          Citations

          This article has been cited 11 times.
          1. Farhadi R, Mohammadi Ghanatghestani AH, Abbasi R, Farshidfar D, Najafi A. In vitro assessment of lavender ethanolic extract supplemented semen extender on cryopreserved ram epididymal sperm quality and fertility outcome. BMC Vet Res 2025 Jun 9;21(1):407.
            doi: 10.1186/s12917-025-04852-3pubmed: 40490767google scholar: lookup
          2. Medica AJ, Gibb Z, Aitken RJ. Optimizing equine sperm quality: an alternative to single layer centrifugation for sperm isolation. Reprod Fertil 2024 Oct 1;5(4).
            doi: 10.1530/RAF-23-0081pubmed: 39437190google scholar: lookup
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            doi: 10.3390/ani14172465pubmed: 39272250google scholar: lookup
          4. Mohammadi T, Hosseinchi Gharehaghaji M. The influence of rutin and chlorogenic acid on oxidative stress and in vivo fertility: Evaluation of the quality and antioxidant status of post-thaw semen from Azari water buffalo bulls. Vet Med Sci 2024 Sep;10(5):e31548.
            doi: 10.1002/vms3.1548pubmed: 39158970google scholar: lookup
          5. Rečková Z, Filipčík R, Soušková K, Kopec T, Hošek M, Pešan V. The efficiency of different types of extenders for semen cooling in stallions. Anim Biosci 2022 May;35(5):670-676.
            doi: 10.5713/ab.21.0300pubmed: 34991206google scholar: lookup
          6. Hassan SA, Khalil WA, Hassan MAE, Yousif AI, Sabry OM, Wink M, Sobeh M. Antioxidant and Antiapoptotic Effects of a Turraea fischeri Leaf Extract on Cryopreserved Goat Sperm. Animals (Basel) 2021 Sep 29;11(10).
            doi: 10.3390/ani11102840pubmed: 34679861google scholar: lookup
          7. Catalán J, Papas M, Trujillo-Rojas L, Blanco-Prieto O, Bonilla-Correal S, Rodríguez-Gil JE, Miró J, Yeste M. Red LED Light Acts on the Mitochondrial Electron Chain of Donkey Sperm and Its Effects Depend on the Time of Exposure to Light. Front Cell Dev Biol 2020;8:588621.
            doi: 10.3389/fcell.2020.588621pubmed: 33365309google scholar: lookup
          8. Saadeldin IM, Khalil WA, Alharbi MG, Lee SH. The Current Trends in Using Nanoparticles, Liposomes, and Exosomes for Semen Cryopreservation. Animals (Basel) 2020 Dec 3;10(12).
            doi: 10.3390/ani10122281pubmed: 33287256google scholar: lookup
          9. Kowalczyk A, Czerniawska-Piątkowska E, Kuczaj M. Factors Influencing the Popularity of Artificial Insemination of Mares in Europe. Animals (Basel) 2019 Jul 19;9(7).
            doi: 10.3390/ani9070460pubmed: 31331026google scholar: lookup
          10. Buss T, Aurich J, Aurich C. Evaluation of a portable device for assessment of motility in stallion semen. Reprod Domest Anim 2019 Mar;54(3):514-519.
            doi: 10.1111/rda.13390pubmed: 30592335google scholar: lookup
          11. Gibb Z, Aitken RJ. The Impact of Sperm Metabolism during In Vitro Storage: The Stallion as a Model. Biomed Res Int 2016;2016:9380609.
            doi: 10.1155/2016/9380609pubmed: 26881234google scholar: lookup
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