Optimal inclusion level of butylated hydroxytoluene in semen extender improves the quality of post-thawed canine sperm

Ziaullah, M.; Ahmad, Irfan; Aleem, M.; Mahmood, Abid; Rehman, Habib; Bhatti, Shaukat Ali; Farooq, Umer; Sohail, Muhammad Umar

doi:10.17221/6273-cjas

Cited by 11 publications

(7 citation statements)

References 26 publications

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“…Concerning the influence of BHT on the functional competence of frozen-thawed dog spermatozoa, the current study indicated that inclusion of 1mM BHT in Tris based extenders caused a significant improvement in motility, viability, acrosomal integrity, plasma membrane integrity as well as sperm DNA integrity. These results are in agreement with a previous study of for Neagu et al (2010); Sahashi et al (2011) and Ziaullah et al (2012) who reported that BHT was found to improve significantly all postthawed canine semen quality parameters in terms of sperm motility, acrosomal integrity, hypo-osmotic swelling response, membrane integrity and viability at an inclusion level of 1mM in the extended semen. BHT has been used successfully for preservation of liquid semen in turkey tom (Donoghue and Donoghue, 1997) and to minimize cryoinjury in ram (Watson and Anderson, 1983), boar (Roca et al, 2004), cattle bull (Shoae and Zamiri, 2008), goat spermatozoa (Khalifa et al, 2008 andNaijian et al, 2013) and human spermatozoa (Merino et al, 2015).…”

Section: Discussionsupporting

confidence: 93%

“…BHT has been used successfully for minimizing cryoinjury in ram (Watson and Anderson, 1983), boar (Roca et al, 2004), cattle bull (Shoae and Zamiri, 2008), and goat spermatozoa (Khalifa et al, 2008). However, there are scanty reports (Neagu et al, 2010;Sahashi et al, 2011 andZiaullah et al, 2012) regarding its cryoprotective potentiality and optimal inclusion level in canine semen. Also, from the best membrane stabilizing substance is Equex STM paste when adding to semen extenders in 0.5% just before freezing (Schäfer-Somi et al, 2006) has shown beneficial effects on frozen-thawed dog semen with regard to post-thaw motility, thermoresistance, increased longevity and plasma membrane integrity (Rota et al, 1997;Peña and Linde-Forsberg 2000b).…”

Section: Introductionmentioning

confidence: 99%

“…Res., 374: 261-268. Ijaz, A.;Hussain, A.;Aleem, M.;Yousaf, M.S. and Rehman, M. (2009): Butylated hydroxytoluene inclusion in semen extender improves the postthawed semen quality of Nili-Ravi buffalo (Bubalus bubalis).…”

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confidence: 99%

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Effect of Adding Equex STM Paste and BHT to Different Extenders on Viability, Plasma Membrane and Dna Integrity of German Shepherd Dog Spermatozoa During Freezing and Thawing Methods in Egypt

2015

Assiut Veterinary Medical Journal

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Many factors influencing semen quality through all steps of cryopreservation such as semen processing, the combination of extender, cooling rate during the freezing procedure and the thawing technique. The present study was conducted to evaluate the potential cryoprotective effect of Equex STM paste and / or butylated hydroxytoluene (BHT) occlusion in Tris and milk based extenders, freezing according to 2 methods, thawing at 2 regimes, and the interactions between these treatments, on the post-thaw semen quality. A total number of 80 ejaculates was collected from 15 German shepherd stud dogs. Semen samples was frozen in two different extenders, Tris (TCF) and milk (SMG) based extenders, with or without 0.5% Equex STM paste and / or 1mM BHT compared with commercial bovine tris based semen extender (BULLXcell ™ , IMV technologies, Franc) using 2 freezing protocols (4 and 6 cm) above LN 2 and 2 thawing rates (45 o C for 30 sec. and 55 o C for 5 sec.). Post-thaw motility, viability, plasma membrane integrity in terms of post-thaw live spermatozoa, acrosomal defects, acrosome reaction and rate of increase in activity of extracellular AST, ALT and ALP enzymes as well as assessment of sperm DNA integrity using comet assay were recorded. The results clarified that addition of Equex STM paste and / or BHT to the TCF and SMG extenders significantly (P<0.01) improved post-thaw motility, viability, plasma membrane integrity and reduced acrosomal defects, acrosomal reaction, release of extracellular AST, ALT and ALP enzymes and DNA damage. The best post-thaw semen qualities was observed when freezing in a Styrofoam box above 6 cm highest above LN 2 and thawing at 55 o C for 5 sec. the superior results were obtained with Tris based extenders than milk based extenders. It is concluded that fortification of 0.5% Equex STM paste and / or 1mM BHT in Tris based extenders freezing above 6 cm highest above LN 2 and thawing at 55 o C for 5 sec. can improve the post-thaw dog semen motility, viability and plasma membrane integrity. Also, the commercial bovine extender BULLXcell™ is useful for cryopreservation of dog semen.

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Section: Discussionsupporting

confidence: 93%

Section: Introductionmentioning

confidence: 99%

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Effect of Adding Equex STM Paste and BHT to Different Extenders on Viability, Plasma Membrane and Dna Integrity of German Shepherd Dog Spermatozoa During Freezing and Thawing Methods in Egypt

2015

Assiut Veterinary Medical Journal

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“…Excessive ROS production during cryopreservation and after thawing not only degrade cellular membranes but may also damage DNA [4], further enhancing dysfunction [5]. To reduce oxidative stress, exogenous antioxidants are frequently added to cryopreservative solutions to sustain semen quality prior to AI [6]. Several studies have confirmed ROS scavenging capacity of various medicinal plant extracts to improve sperm motility and increase fertility rates [7][8][9][10].…”

Section: Introductionmentioning

confidence: 99%

Antioxidant and Antiapoptotic Effects of a Turraea fischeri Leaf Extract on Cryopreserved Goat Sperm

Hassan

Khalil

Hassan

et al. 2021

Animals

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This study evaluated the efficacy of Turraea fischeri leaf extract for maintaining the viability of cryopreserved goat sperm. Ejaculated semen was collected from 5 mature Baladi bucks (50–60 kg, 2–4 years of age) and those samples with mass motility ≥ 70% and sperm concentration ≥ 2.5 × 109/mL were selected, pooled, and divided into 4 aliquots. Each aliquot was diluted in Tris-citric-soybean lecithin extender containing a different concentration of T. fischeri leaf extract (0, 125, 250, or 375 µg/mL). Treated semen samples were cooled to 5 °C, transferred to 0.25-mL French straws, and stored in liquid nitrogen (LN2) at −196 °C. After thawing, membrane integrity was examined by transmission electron microscopy, apoptotic activity by Annexin/propidium iodide staining and flow cytometry, and both enzyme activities and antioxidant capacity by spectroscopic assays. The leaf extract at 375 µg/mL significantly improved semen quality as indicated by enhanced total antioxidant capacity, reduced H2O2 concentration, a greater proportion of structurally intact motile sperm, and concomitant reductions in apoptosis and necrosis. The extract also significantly increased the proportion of sperm with a contiguous plasma membrane and intact acrosome (p < 0.05). Furthermore, LC-MS revealed numerous secondary metabolites in the extract that may contribute to sperm cryopreservation.

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“…The LPO process caused by ROS (H 2 O 2 ) is detrimental to sperm survivability. As a result of high contents of polyunsaturated fatty acids in the plasma membrane and lack of antioxidant enzymes, mammalian spermatozoa are susceptible to LPO induced damage and loss of sperm functions [27,28]. Increasing ROS generation under oxidative stress (OS) leads to increased sperm plasma membrane failure, damaged spermatozoa [29], reduced sperm cell cytoplasm [30], and finally a marked reduction in viability, the integrity of the sperm membrane, and fertilizing ability and increased damage to sperm DNA [31].…”

Section: Introductionmentioning

confidence: 99%

The Current Trends in Using Nanoparticles, Liposomes, and Exosomes for Semen Cryopreservation

Saadeldin

Khalil

Alharbi

et al. 2020

Animals

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Cryopreservation is an essential tool to preserve sperm cells for zootechnical management and artificial insemination purposes. Cryopreservation is associated with sperm damage via different levels of plasma membrane injury and oxidative stress. Nanoparticles are often used to defend against free radicals and oxidative stress generated through the entire process of cryopreservation. Recently, artificial or natural nanovesicles including liposomes and exosomes, respectively, have shown regenerative capabilities to repair damaged sperm during the freeze–thaw process. Exosomes possess a potential pleiotropic effect because they contain antioxidants, lipids, and other bioactive molecules regulating and repairing spermatozoa. In this review, we highlight the current strategies of using nanoparticles and nanovesicles (liposomes and exosomes) to combat the cryoinjuries associated with semen cryopreservation.

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Optimal inclusion level of butylated hydroxytoluene in semen extender improves the quality of post-thawed canine sperm

Cited by 11 publications

References 26 publications

Effect of Adding Equex STM Paste and BHT to Different Extenders on Viability, Plasma Membrane and Dna Integrity of German Shepherd Dog Spermatozoa During Freezing and Thawing Methods in Egypt

Effect of Adding Equex STM Paste and BHT to Different Extenders on Viability, Plasma Membrane and Dna Integrity of German Shepherd Dog Spermatozoa During Freezing and Thawing Methods in Egypt

Antioxidant and Antiapoptotic Effects of a Turraea fischeri Leaf Extract on Cryopreserved Goat Sperm

The Current Trends in Using Nanoparticles, Liposomes, and Exosomes for Semen Cryopreservation

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