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Tropical animal health and production2026; 58(5); 326; doi: 10.1007/s11250-026-05136-0

Effect of Spirulina platensis extract (C-phycocyanin) supplementation to the semen extender on kinematic and oxidative parameters of Marwari stallions during freezing.

Abstract: The current research aimed at examining the impact of different quantities of Spirulina platensis extract (C-phycocyanin) supplemented to the semen extender on the post-thaw quality of Marwari stallion spermatozoa. Semen samples were obtained via an artificial vagina from six fertile stallions and diluted with an extender containing Spirulina platensis extract (C-phycocyanin) at concentrations of 10, 20, and 50 µg/mL, or without Spirulina as a control. Aliquots of diluted semen were subjected to cooling (4 °C, 120 min) and then frozen (-196 °C, 1 day), after which the kinematic parameters (VAP - average path velocity, VCL - curvilinear velocity, VSL - straight line velocity, STR - straightness, and LIN - linearity) as well as antioxidant activities of superoxide dismutase (SOD), catalase (CAT), total antioxidant capacity (T-AOC), and malondialdehyde (MDA) were assessed in cryopreserved spermatozoa. The findings showed that Spirulina, when combined with the extender, significantly enhanced (P < 0.05) sperm kinematic characteristics and antioxidant activities post-freezing. Nonetheless, the incorporation of 50 µg C-phycocyanin/150 million spermatozoa into the freezing extender enhanced (P < 0.05) the motility metrics and antioxidant activities of frozen/thawed spermatozoa. The current findings indicated that the inclusion of 50 µg C-phycocyanin/150 million spermatozoa in the freezing extender enhanced semen quality and decreased cryodamage to Marwari stallion spermatozoa.
© 2026. The Author(s), under exclusive licence to Springer Nature B.V.
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Publication Date: 2026-06-12 PubMed ID: 42283944PubMed Central: 6431823DOI: 10.1007/s11250-026-05136-0Google 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 study investigated how adding different amounts of Spirulina platensis extract (C-phycocyanin) to the semen extender affects the quality and antioxidant properties of frozen-thawed sperm from Marwari stallions.

Research Objective and Methodology

  • Purpose: To evaluate the effect of Spirulina platensis extract supplementation on the kinematic (movement) and oxidative (antioxidant) parameters of Marwari stallion sperm after freezing and thawing.
  • Subjects: Semen collected from six fertile Marwari stallions using an artificial vagina.
  • Treatments: Semen diluted with extender containing varying concentrations of Spirulina extract (C-phycocyanin) at 10, 20, and 50 µg/mL, and a control group without Spirulina.
  • Procedure: The diluted semen was cooled at 4°C for 120 minutes, then frozen at -196°C for 1 day.
  • Post-thaw analysis: Assessed sperm movement characteristics and antioxidant enzyme activities.

Sperm Kinematic Parameters Evaluated

  • VAP (Average Path Velocity): The average velocity of sperm along its average path.
  • VCL (Curvilinear Velocity): The velocity along the actual curved sperm path.
  • VSL (Straight Line Velocity): The velocity along a straight line from the start to the end point.
  • STR (Straightness): The ratio of VSL to VAP, indicating how linear the sperm movement is.
  • LIN (Linearity): Ratio of VSL to VCL, representing the linearity of the sperm trajectory.

Oxidative Parameters Measured

  • Superoxide Dismutase (SOD): Enzyme that neutralizes superoxide radicals.
  • Catalase (CAT): Enzyme that converts hydrogen peroxide into water and oxygen, reducing oxidative stress.
  • Total Antioxidant Capacity (T-AOC): Overall ability of antioxidants present in the sample to counteract oxidative damage.
  • Malondialdehyde (MDA): Indicator of lipid peroxidation and oxidative damage to sperm membranes.

Key Findings

  • Supplementation of Spirulina platensis extract to the semen extender significantly improved sperm kinematic parameters after thawing (P < 0.05), indicating enhanced sperm motility and movement quality.
  • Antioxidant enzyme activities (SOD, CAT) and total antioxidant capacity increased in treated groups compared to the control, suggesting better protection against oxidative damage during the freeze-thaw process.
  • The highest Spirulina concentration tested (50 µg C-phycocyanin per 150 million spermatozoa) provided the most significant improvement in both motility and antioxidant parameters after thawing.
  • The level of malondialdehyde (MDA), which signals membrane damage, was reduced with Spirulina supplementation, indicating decreased cryodamage to sperm membranes.

Conclusions and Implications

  • Adding 50 µg of C-phycocyanin extract from Spirulina platensis to semen extenders enhances the post-thaw quality of Marwari stallion sperm.
  • This supplementation improves sperm motility and antioxidant defense, thereby reducing the harmful effects of oxidative stress caused by freezing and thawing.
  • The results suggest that Spirulina platensis extract could be a beneficial additive in cryopreservation protocols to maintain stallion sperm function and fertility.

Cite This Article

APA
Yadav V, Talluri TR, Dholpuria S, Khandelwal S, Joshi A. (2026). Effect of Spirulina platensis extract (C-phycocyanin) supplementation to the semen extender on kinematic and oxidative parameters of Marwari stallions during freezing. Trop Anim Health Prod, 58(5), 326. https://doi.org/10.1007/s11250-026-05136-0

Publication

Tropical animal health and production
ISSN: 1573-7438
NlmUniqueID: 1277355
Country: United States
Language: English
Volume: 58
Issue: 5
PII: 326

Researcher Affiliations

Yadav, Vishal
  • Department of Veterinary Gynaecology and Obstetrics, College of Veterinary and Animal Science, Bikaner, Rajasthan University of Veterinary and Animal Sciences, Bikaner, Rajasthan, 334001, India. vishalyadav1131997@gmail.com.
Talluri, Thirumala Rao
  • Equine Production Campus, ICAR-National Research Centre on Equines, Bikaner, Rajasthan, 334001, India.
Dholpuria, Sandeep
  • Department of Veterinary Gynaecology and Obstetrics, College of Veterinary and Animal Science, Bikaner, Rajasthan University of Veterinary and Animal Sciences, Bikaner, Rajasthan, 334001, India.
Khandelwal, Shivali
  • Department of Veterinary Microbiology and Biotechnology, College of Veterinary and Animal Science, Bikaner, Rajasthan University of Veterinary and Animal Sciences, Bikaner, Rajasthan, 334001, India.
Joshi, Abhishek
  • Department of Livestock Production and Management, College of Veterinary and Animal Science, Bikaner, Rajasthan University of Veterinary and Animal Sciences, Bikaner, Rajasthan, 334001, India.

MeSH Terms

  • Animals
  • Male
  • Spirulina / chemistry
  • Phycocyanin / pharmacology
  • Semen Preservation / veterinary
  • Semen Preservation / methods
  • Cryopreservation / veterinary
  • Cryopreservation / methods
  • Horses / physiology
  • Spermatozoa / drug effects
  • Spermatozoa / physiology
  • Cryoprotective Agents / pharmacology
  • Antioxidants
  • Semen / drug effects
  • Sperm Motility / drug effects
  • Semen Analysis / veterinary
  • Freezing

Conflict of Interest Statement

Declarations. Ethical approval: The current study was conducted and approved under the guidelines laid down by the institute’s Animal Ethics Committee (Institutional Animal Ethics Committee, College of Veterinary & Animal Science, Rajasthan University of Veterinary and Animal Sciences, Bikaner Rajasthan, India, Reg. No.- 2044/GO/Re/SL/18/CPCSEA, No.- CVAS/IAEC/2022-23/30). Consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: No competing interests exists.

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