Effects of Costmary (Tanacetum balsamita L. ) extract on selected quality parameters of Moghani ram sperm after cryopreservation

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VAHEDI V.

Title

Effects of Costmary (Tanacetum balsamita L. ) extract on selected quality parameters of Moghani ram sperm after cryopreservation

Author(s)

VAHEDI V. | Issue Writer Certificate

Keywords

Antioxidant

Freeze-thawing

Membrane integrity

Ram semen

Tanacetum balsamita extract

Abstract

Introduction: The artificial insemination is based on sperm cryopreservation technique that induces irreversible damages to motility, viability, plasma Membrane integrity, and fertility of the sperm (Purdy 2006; Baghshahi et al. 2014). Physical and chemical damages during cryopreservation are associated with reactive oxygen species (ROS) and lipid peroxidation of the phospholipids in the membrane by free radicals (Chatterjee et al. 2001). The removal of ROS is catalyzed by Antioxidant enzymes such as glutathione peroxidase (GSH-PX), superoxide dismutase (SOD) and catalase (CAT). Numerous non-enzymatic defenses (vitamin C, vitamin E, and glutathione (GSH)) are also employed to protect cells from oxidative damages. An imbalance between free radical production and their removal leads to ageing and its subsequent progressive damage. Many studies have been investigated the protecting effects of synthetic and natural Antioxidants against oxidative damage on spermatozoa during cryopreservation (Malo et al. 2010). Of many classes of natural compounds, plant secondary metabolites (e. g., flavonoids, tannins, coumarins, xanthons, phenolics, lignans and terpenoids) have repetitively shown antioxidative properties. Correspondingly, several studies have shown the positive effects of herbal Antioxidants on the sperm quality during the freezing-thawing processes (Daghigh Kia et al. 2016; Vahedi et al. 2018). Costmary (Tanacetum balsamita L. ) with Antioxidant effects contains phenolic compounds, such as flavonoids and phenolic acids (Shahhoseini et al. 2019) (Bą czek et al. 2017). The aim of current study was to evaluate the effect of costmary extract as a natural Antioxidant on post-thawed ram sperm quality. Material and methods: This study was performed at the Iranian Moghani sheep Breeding Center located in Jafarabad city, Province of Ardebil, Iran. Four mature and fertile rams (3-4 years old, mean live weight of 70± 4. 2 kg), were used in this study. Ejaculates were collected twice a week for 8 weeks by an artificial vagina (42-43° C). Only samples containing spermatozoa with greater than 70% motility were accepted for the experiment. To eliminate individual differences, semen samples were pooled and processed. The pooled ejaculate was diluted (37 ◦ C) using egg yolk-citrate extender containing different concentrations of Tanacetum balsamita extract (0, 2, 4, 8, 12, and 16 mL/dL). Diluted semen samples were aspirated into 0. 25 ml straws and equilibrated at 4° C for 3 h. After equilibration, the straws were placed on liquid nitrogen (LN2) vapor for 8 min, then plunged into liquid nitrogen, and stored in a liquid nitrogen tank until thawed and used for evaluation of sperm parameters. Prior to analysis, the frozen straws were thawed individually in a water bath (37 ◦ C) for 30 s. A computer-assisted sperm analysis (HFT CASA, Hooshmand Fanavar Tehran Co, Iran) was used to analyze sperm motility and velocity characteristics. Sperm viability was assessed using a modification of the eosin-nigrosin staining method described by Evans and Maxwell (1987). Sperm membrane functionality was evaluated using the hypoosmotic swelling test (HOST) (Revell and Mrode, 1994). For the assessment of the sperm morphology abnormalities, at least three drops of each sample were added to Eppendorf tubes containing 1 ml of Hancock solution (62. 5 ml formalin (37%), 150 ml sodium saline solution, 150 ml buffer solution and 500 ml double-distilled water). The prepared slides were assessed by phase-contrast microscopy at the 400× magnification. All data were analyzed by completely randomized design using the GLM procedure of SAS (version 9. 1, SAS Institute, 2004). Results and discussion: Samples cryopreserved in 8 and 12 mL/dL had higher total motility and progressive motility compared to the control group (P<0. 05). The percentage of VSL, VCL and VAP were higher (P<0. 05) in the extender containing 8 and 12 mL/dL extract compared to control and 16 mL/dL groups. LIN parameter was higher (p<0. 05) in 8 mL/dL compared to 16 mL/dL extract (46. 83± 3. 82% vs. 40. 52± 3. 23%). For parameter STR, the highest value (P<0. 05) was observed at 8 and 12 mL/dL of extract (81. 09± 7. 56% and 80. 27± 7. 18%, respectively). The highest (p<0. 05) percentage of sperm viability and plasma Membrane integrity were observed in groups containing 8 and 12 mL/dL extract. Percentage of acrosome abnormality was higher (p< 0. 05) in 12 mL/dL extract groups (21. 25%) compared with control and 2 mL/dL extract groups (26. 70% and 27. 23%, respectively). Some studies have reported that herbal Antioxidants reduce the free radicals following the freeze– thawing process (Ashrafi et al. 2013). In the present study, costmary extract treatment of the semen resulted in a significant improvement in motility parameters, viability and Membrane integrity of frozen-thawed ram sperm. The main constituents of costmary extract have been reported to be polyphenolic compounds, such as flavonoids and phenolic acids (Faraloni, 2018). Among flavonoids, glycosides of luteolin, apigenin and quercetin are the most abundant ones while phenolic acids are mainly represented by chlorogenic, caffeic and dicaffeoylquinic acids. Reactive oxygen species occurrences during cryopreservation would lead to reduction of oxygen and sperm membrane lipid peroxidation thus loss of its integrity. Flavonoids protect Membrane integrity by preventing the production of free radicals and lipid peroxidation hence hindering oxidative damage to the membrane components (Daghigh Kia et al. 2016). Our findings demonstrate the free radical scavenging and protective effect of costmary extract against oxidative damage during sperm cryopreservation. Conclusion: In conclusion, this study showed that supplementation of extender with 12 mL/dL Tanacetum balsamita L. extract has a beneficial effect on the quality of frozen-thawed Ram semen.

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APA: Copy

VAHEDI, V.. (2021). Effects of Costmary (Tanacetum balsamita L. ) extract on selected quality parameters of Moghani ram sperm after cryopreservation. ANIMAL SCIENCE RESEARCHES (FACULTY OF AGRICULTURE, UNIVERSITY OF TABRIZ), 30(4 ), 71-82. SID. https://sid.ir/paper/957099/en

Vancouver: Copy

VAHEDI V.. Effects of Costmary (Tanacetum balsamita L. ) extract on selected quality parameters of Moghani ram sperm after cryopreservation. ANIMAL SCIENCE RESEARCHES (FACULTY OF AGRICULTURE, UNIVERSITY OF TABRIZ)[Internet]. 2021;30(4 ):71-82. Available from: https://sid.ir/paper/957099/en

IEEE: Copy

V. VAHEDI, “Effects of Costmary (Tanacetum balsamita L. ) extract on selected quality parameters of Moghani ram sperm after cryopreservation,” ANIMAL SCIENCE RESEARCHES (FACULTY OF AGRICULTURE, UNIVERSITY OF TABRIZ), vol. 30, no. 4 , pp. 71–82, 2021, [Online]. Available: https://sid.ir/paper/957099/en

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Scientific Information Database

ISSN: 2588-4824

Journal Paper

Effects of Costmary (Tanacetum balsamita L. ) extract on selected quality parameters of Moghani ram sperm after cryopreservation