This study was designed to assess the effects of using tris-soybean lecithin (TSL)-based extender supplemented with bovine serum albumin (BSA) on the quality of ram epididymal spermatozoa during refrigerated storage.
Method:
Epididymal sperm were collected from 22 Zandi rams, diluted in TSL-based extender at different concentrations (0%, 2.5%, 5%, 7.5%, and 10%) of BSA, and stored for 5 days at 4°C. Sperm parameters including motility, viability, plasma membrane integrity, chromatin protamination, and malondialdehyde (MDA) content were evaluated at 0, 24, 72, and 120 hours of refrigeration.
Results:
The addition of 10% BSA to the extender significantly improved sperm viability at 24 and 120 hours of refrigerated liquid storage (p < 0.05). An enhancement in plasma membrane integrity was observed along with a decrease in MDA level by increasing the concentration of BSA from 0% to 10% (p > 0.05). Sperm motion characteristics were higher in the BSA-free group at 120 hours of preservation (p < 0.05). No statistical difference was found for nuclear protamination between experimental groups (p > 0.05).
Conclusion:
BSA supplementation in TSL-based extender can preserve the viability of epididymal ram spermatozoa during liquid storage at 4°C.
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References
1.
Tamayo-CanulJ, ÁlvarezM, Mata-CampuzanoM, et al.Effect of storage method and extender osmolality in the quality of cryopreserved epididymal ram spermatozoa. Anim Reprod Sci, 2011; 129:188–199.
2.
BertolMAF. Cryopreservation of Epididymal Sperm. In: Marco-Jiménez F, Akdemir H, eds. Cryopreservation in Eukaryotes. London: IntechOpen 2016:121–135.
3.
Tamayo-CanulJ, AlvarezM, López-UrueñaE, et al.Undiluted or extended storage of ram epididymal spermatozoa as alternatives to refrigerating the whole epididymes. Anim Reprod Sci, 2011; 126:76–82.
4.
ÁlvarezM, Tamayo-CanulJ, Martínez-RodríguezC, et al.Specificity of the extender used for freezing ram sperm depends of the spermatozoa source (ejaculate, electroejaculate or epididymis). Anim Reprod Sci, 2012; 132:145–154.
5.
KaabiM, PazP, AlvarezM, et al.Effect of epididymis handling conditions on the quality of ram spermatozoa recovered post-mortem. Theriogenology, 2003; 60:1249–1259.
6.
Fernández AbellaD, Da CostaM, GuérinY, et al.Fertility of undiluted ram epididymal spermatozoa stored for several days at 4°C. Animal, 2015; 9:313–319.
7.
AnelL, AlvarezM, Martinez-PastorF, et al.Improvement strategies in ovine artificial insemination. Reprod Domest Anim, 2006; 41Suppl 2:30–42.
Fernández-SantosM, Domínguez-RebolledoA, EstesoM, et al.Refrigerated storage of red deer epididymal spermatozoa in the epididymis, diluted and with vitamin C supplementation. Reprod Domest Anim, 2009; 44:212–220.
10.
SarıözkanS, TürkG, CantürkF, et al.The effect of bovine serum albumin and fetal calf serum on sperm quality, DNA fragmentation and lipid peroxidation of the liquid stored rabbit semen. Cryobiology, 2013; 67:1–6.
11.
ZhangX-G, YanG-J, HongJ-Y, et al.Effects of bovine serum albumin on boar sperm quality during liquid storage at 17°C. Reprod Domest Anim, 2015; 50:263–269.
12.
PaulenzH, SöderquistL, Pérez-PéR, et al.Effect of different extenders and storage temperatures on sperm viability of liquid ram semen. Theriogenology, 2002; 57:823–836.
13.
BucakMN, TekinN. Protective effect of taurine, glutathione and trehalose on the liquid storage of ram semen. Small Rumin Res, 2007; 73:103–108.
14.
BucakMN, BoduM, BaşpinarN, et al.Influence of ellagic acid and ebselen on sperm and oxidative stress parameters during liquid preservation of ram semen. Cell J, 2019; 21:7–13.
15.
RastegarniA, ShetabiN, Rezaei TopT, et al.Effects of exogenous glutathione supplementation in Biocell® extender on quality of cryopreserved buffalo (Bubalus bubalis) semen. J Anim Vet Adv, 2012; 11:3437–3443.
16.
El-KonI. Testing usability of bovine serum albumin (BSA) for preservation of Egyptian buffalo semen. Am J Agric Environ Sci, 2011; 11:495–502.
17.
RahimizadehP, TobraggalehTR, EsmaeiliV, et al.Effects of bovine serum albumin on ram epididymal spermatozoa during refrigerated storage. Cryobiology, 2018; 80:189.
18.
UysalO, BucakMN. Effects of oxidized glutathione, bovine serum albumin, cysteine and lycopene on the quality of frozen-thawed ram semen. Acta Vet Brno, 2007; 76:383–390.
19.
RocheM, RondeauP, SinghNR, et al.The antioxidant properties of serum albumin. FEBS Lett, 2008; 582:1783–1787.
20.
BlankM, SooL, BrittenJS. Adsorption of albumin on rabbit sperm membranes. J Membr Biol, 1976; 29:401–409.
21.
MatsuokaT, ImaiH, KohnoH, et al.Effects of bovine serum albumin and trehalose in semen diluents for improvement of frozen-thawed ram spermatozoa. J Reprod Dev, 2006; 52:675–683.
22.
FukuiY, KohnoH, TogariT, et al.Fertility of ewes inseminated intrauterinally with frozen semen using extender containing bovine serum albumin. J Reprod Dev, 2007; 53:959–962.
23.
SariözkanS, TuncerPB, BucakMN, et al.Influence of various antioxidants on microscopic-oxidative stress indicators and fertilizing ability of frozen-thawed bull semen. Acta Vet Brno, 2009; 78:463–469.
24.
EmamverdiM, ZhandiM, ShahnehAZ, et al.Flow cytometric and microscopic evaluation of post-thawed ram semen cryopreserved in chemically defined home-made or commercial extenders. Anim Prod Sci, 2015; 55:551.
25.
RahimizadehP, TopraggalehTR, Nasr-EsfahaniMH, et al.The alteration of PLCζ protein expression in unexplained infertile and asthenoteratozoospermic patients: A potential effect on sperm fertilization ability. Mol Reprod Dev, 2020; 87:115–123.
26.
ZiaratiN, TopraggalehTR, RahimizadehP, et al.Micro-quantity straw as a carrier for cryopreservation of oligozoospermic semen samples: Effects of storage times and cryoprotectant. Cryobiology, 2019; 86:65–70.
27.
SelvarajuS, RajuP, RaoSBN, et al.Evaluation of maize grain and polyunsaturated fatty acid (PUFA) as energy sources for breeding rams based on hormonal, sperm functional parameters and fertility. Reprod Fertil Dev, 2012; 24:669.
28.
AzawiOI, HusseinEK. Study on the effect of adding bovine serum albumin to semen diluent on the viability of awassi ram semen preserved at 5°C. J Adv Vet Res, 2011; 1:115–118.
29.
ForouzanfarM, SharafiM, HosseiniSM, et al.In vitro comparison of egg yolk–based and soybean lecithin–based extenders for cryopreservation of ram semen. Theriogenology, 2010; 73:480–487.
30.
EmamverdiM, ZhandiM, Zare ShahnehA, et al.Optimization of ram semen cryopreservation using a chemically defined soybean lecithin-based extender. Reprod Domest Anim, 2013; 48:899–904.
31.
ScottT, VoglmayrJ, SetchellB. Lipid composition and metabolism in testicular and ejaculated ram spermatozoa. Biochem J, 1967; 102:456–461.
32.
WhiteIG, WalesRG. Comparison of epididymal and ejaculated semen of the ram. J Reprod Fertil, 1961; 2:225–237.
33.
OsmanK, NangC, IbrahimS, et al.Albumin improved spermatozoa quality and DNA integrity for freezing-free preservation. Int J Biol Med Res, 2012; 3:1670–1679.
34.
SöderquistL, Madrid-BuryN, Rodriguez-MartinezH. Assessment of ram sperm membrane integrity following different thawing procedures. Theriogenology, 1997; 48:1115–1125.
35.
MaxwellWMC, WatsonPF. Recent progress in the preservation of ram semen. Anim Reprod Sci, 1996; 42:55–65.
36.
MaxwellW, StojanovT. Liquid storage of ram semen in the absence or presence of some antioxidants. Reprod Fertil Dev, 1996; 8:1013.
37.
GökçeE, AlçayS, GülZ. Positive effect of BSA supplemented soybean lecithin based extender on liquid storage of ram semen at 5°C. Ankara Üniversitesi Vet Fakültesi Derg, 2017; 64:313–320.
38.
AbdillahDA, SetyawanEMN, OhHJ, et al.Iodixanol supplementation during sperm cryopreservation improves protamine level and reduces reactive oxygen species of canine sperm. J Vet Sci, 2019; 20:79.
39.
FloresE, Ramió-LluchL, BucciD, et al.Freezing-thawing induces alterations in histone H1-DNA binding and the breaking of protein-DNA disulfide bonds in boar sperm. Theriogenology, 2011; 76:1450–1464.
40.
PerisSI, BilodeauJ-F, DufourM, et al.Impact of cryopreservation and reactive oxygen species on DNA integrity, lipid peroxidation, and functional parameters in ram sperm. Mol Reprod Dev, 2007; 74:878–892.
41.
PerreaultSD, BarbeeRR, ElsteinKH, et al.Interspecies differences in the Stability of mammalian sperm nuclei assessed in vivo by sperm microinjection and in vitro by flow cytometry. Biol Reprod, 1988; 39:157–167.
