Abstract
This study aimed to test different doses of L-glutamine and trehalose in the canine semen diluent while determining their protective effects on spermatological and biochemical indices of the thawed samples. Semen samples were collected from three fertile dogs using the digital manipulation method. The mixed ejaculates were divided into five portions at 37 °C and diluted with additives. Five study groups were formed with L-glutamine (10 and 20 mM), trehalose (25 and 50 mM), and no additives (control). After the dilution, the semen samples were cooled for 1.5 h at 5 °C and frozen (-110 to -120 °C) in liquid nitrogen vapor. Then, they were stored at -196 °C. For spermatological evaluations, samples were thawed at 38 °C for 30 s. L-glutamine (20 mM) was found to be significantly different (P < 0.05) and led to higher percentages of motility, membrane integrity, and acrosome integrity compared to the control group. Considering the total oxidant status (TOS) assay, the lower values were determined in all the antioxidant groups compared to the control group (P < 0.05). Supplementing the semen extender with L-glutamine showed a higher total antioxidant status (TAS) concentration compared to the control group (P < 0.05). As a result of this study, a higher protective effect was found in all the spermatological evaluations after thawing the frozen semen samples, especially in the group containing L-glutamine (20 mM).
Highlights
This study aimed to test different doses of L-glutamine and trehalose in the canine semen diluent while determining their protective effects on spermatological and biochemical indices of the thawed samples
The highest total antioxidant status (TAS) concentrations were reached in the 20 mM glutamine group, which showed a significant difference compared to the control group
We found the 50 mM trehalose group to be useful in the total oxidant status (TOS) value (Table 2) and it showed a significant difference compared to the control group (P < 0.05)
Summary
This study aimed to test different doses of L-glutamine and trehalose in the canine semen diluent while determining their protective effects on spermatological and biochemical indices of the thawed samples. As a result of this study, a higher protective effect was found in all the spermatological evaluations after thawing the frozen semen samples, especially in the group containing L-glutamine (20 mM). To reduce the negative effects of oxidative stress on semen and to protect the spermatological indices, many studies have documented that adding antioxidants to the extender could be beneficial in preserving the sperm indices after thawing in animals such as rams (Gungor et al 2018), bulls (Büyükleblebici et al 2014), and canines (Caturla-Sánchez et al 2018). The post-thaw semen analysis determined that trehalose preserved motility, vitality, and membrane integrity in different species while reducing oxidative stress (Iqbal et al 2016; Park et al 2018; Keskin et al 2020)
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