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RESEARCH ARTICLE
Contents Vol 64(11)

Effect of L-carnosine on frozen ram-semen quality evaluated by CASA and flow-cytometry

İbrahim Halil Güngör https://orcid.org/0000-0002-5250-1478 A * , Seyfettin Gür https://orcid.org/0000-0003-0096-2501 A and Edanur Güler Ekmen https://orcid.org/0000-0001-8473-7592 B
+ Author Affiliations
- Author Affiliations

A Department of Reproduction and Artificial Insemination, Fırat University, Faculty of Veterinary Medicine, Elazığ, Türkiye.

B Department of Physiology, Fırat University, Faculty of Veterinary Medicine, Elazığ, Türkiye.

* Correspondence to: ihgungor@firat.edu.tr

Handling Editor: Rodolfo Ungerfeld

Animal Production Science 64, AN24048 https://doi.org/10.1071/AN24048
Submitted: 16 February 2024  Accepted: 3 July 2024  Published: 29 July 2024

© 2024 The Author(s) (or their employer(s)). Published by CSIRO Publishing

Abstract

Context

Successful freezing of ram semen has not yet reached the desired levels. The main reason for this situation could be due to the fact that the spermatozoa of this species have a lipid composition different from that of other species.

Aims

The objective of the study was to evaluate the effect of different concentrations of L-carnosine added to the extender on ram semen after being frozen and thawed.

Methods

Semen was collected from six Akkaraman rams twice a week for a period of 3 weeks. Pooling was performed at each time. The semen were reconstituted with a pre-prepared tris + egg yolk solution and different amounts of L-carnosine to form experimental groups (Group 1: 1 mM, Group 2: 5 mM, Group 3: 10 mM, Group 4: 20 mM, Group 5: control) and were drawn into 0.25 mL mini straws. Subsequently, the samples were subjected to freezing by using an automated freezing device. Following the freezing process, the straws were placed in containers containing liquid nitrogen and thawed after 24 h.

Key results

After thawing, it was found that the samples containing 5 mM L-carnosine had superior results in all analyses. This concentration exhibited significantly higher percentages of progressive, total, and rapid sperm motility, live spermatozoa, high mitochondrial membrane potential rate, and higher GSH-Px concentrations. In addition, it was determined that 5 mM L-carnosine group protected the membrane integrity and significantly decreased the rate of abnormal spermatozoa, acrosomal damage rate, low mitochondrial membrane potential and apoptotic cell rate.

Conclusions

As a result, It was determined that adding 5 mM of L-carnosine to the semen extender during the freezing of ram samples would be beneficial for successful freezing.

Implications

The addition of 5 mM L-carnosine to ram-semen extenders ensures the freezability of the semen of this species; thus, this protocol could be used to perform artificial insemination with frozen ram semen.

Keywords: apoptosis, flow-cytometer, freeze-thawing, L-carnosine, oxidative stress, pH, ram, semen.

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