This study aimed to evaluate the viability of gametes in zebrafish (Danio rerio), at different rigor mortis stages. Viability assessments were conducted on oocytes at various developmental stages using LIVE/DEAD and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. For sperm evaluation, both kinetic (computer-assisted sperm analysis) and morphological assessments (Rose Bengal staining) were performed. Results demonstrated that rigor mortis progression significantly impacted oocyte viability during post-rigor stages, with the following viability rates: pre-rigor (70.43 ± 12.31%), fresh/control (46.43 ± 12.54%), post-rigor (27.62 ± 22.29%), and rigor mortis (comparable to fresh/control). Conversely, sperm kinetics exhibited nuanced responses to the rigor mortis stages, with specific parameters showing sensitivity, whereas the others remained relatively stable. Sperm motility was higher in the fresh/control (63.23 ± 19.03%) and pre-rigor (58.96 ± 14.38%) compared to the post-rigor group (3.34 ± 4.65%). This study highlights the significance of the pre-rigor for successful gamete collection and preservation. These findings provide valuable insights for conservation efforts and optimization of genetic resource management for endangered fish species. This study aimed to develop effective assistive reproductive techniques by elucidating the interplay between rigor mortis and gamete quality, contributing to the broader goals of species conservation and maintenance of genetic diversity in fish populations.
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References
1.
ShinoharaT, InoueK, OgonukiN, et al.Birth of offspring following transplantation of cryopreserved immature testicular pieces and in-vitro microinsemination. Hum Reprod, 2002; 17(12):3039–3045; doi: 10.1093/humrep/17.12.3039
2.
TharasanitT, BuarpungS, Manee-InS, et al.Birth of kittens after the transfer of frozen-thawed embryos produced by intracytoplasmic sperm injection with spermatozoa collected from cryopreserved testicular tissue. Reprod Domest Anim, 2012; 47(Suppl 6):305–308; doi: 10.1111/rda.12072
3.
OctaveraA, YoshizakiG. Production of Chinese rosy bitterling offspring derived from frozen and vitrified whole testis by spermatogonial transplantation. Fish Physiol Biochem, 2020; 46(4):1431–1442; doi: 10.1007/s10695-020-00802-y
4.
KoteeswaranR, PandianTJ. Live sperm from post-mortem preserved Indian catfish. Current Science [Internet], 2002; 82(4):447–450. https://www.jstor.org/stable/24106659
5.
Umaa RaniK, DhanasekarK, MunuswamyN. Fertilizability of cryopreserved and cadaveric fish spermatozoa of freshwater catfish Pangasius sutchi (Fowler, 1937). Aquac Res, 2016; 47(5):1511–1518; doi: 10.1111/are.12611
6.
KirankumarS, PandianTJ. Interspecific androgenetic restoration of rosy barb using cadaveric sperm. Genome, 2004; 47(1):66–73; doi: 10.1139/g03-104
7.
YangF, IchidaK, YoshizakiG. Gametogenesis commencement in recipient gonads using germ cells retrieved from dead fish. Aquaculture, 2022; 552:737952; doi: 10.1016/j.aquaculture.2022.737952
8.
AliS, AaldersJ, RichardsonMK. Teratological effects of a panel of sixty water-soluble toxicants on zebrafish development. Zebrafish, Larchmont, 2014; 11(2):129–141; doi: 10.1089/zeb.2013.0901
9.
CachatJ, CanavelloP, EleganteM, et al.Modeling withdrawal syndrome in zebrafish. Behav Brain Res, 2010; 208(2):371–376; doi: 10.1016/j.bbr.2009.12.004
10.
LawrenceC. The husbandry of zebrafish (Danio rerio): A review. Aquaculture, 2007; 269(1–4):1–20; doi: 10.1016/j.aquaculture.2007.04.077
11.
HoweK, ClarkMD, TorrojaCF, et al.The zebrafish reference genome sequence and its relationship to the human genome. Nature, 2013; 496(7446):498–503; doi: 10.1038/nature12111
12.
YeM, ChenY. Zebrafish as an emerging model to study gonad development. Comput Struct Biotechnol J, 2020; 18:2373–2380; doi: 10.1016/j.csbj.2020.08.025
13.
WaghmareSG, SamarinAM, FraněkR, et al.Oocyte Ageing in Zebrafish Danio rerio (Hamilton, 1822) and its consequence on the viability and ploidy anomalies in the progeny. Animals (Basel), 2021; 11(3):912; doi: 10.3390/ani11030912
14.
Cardona-CostaJ, Pérez-CampsM, García-XiménezF, EspinósFJ. Effect of Gametes Aging on Their Activation and Fertilizability in Zebrafish (Danio rerio). Zebrafish, 2009; 6(1):93–95; doi: 10.1089/zeb.2008.0578
15.
Percie Du SertN, HurstV, AhluwaliaA, et al.The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research. Boutron I, editor. PLoS Biol, 2020; 18(7):e3000410; doi: 10.1371/journal.pbio.3000410
16.
MatthewsM, VargaZM. Anesthesia and euthanasia in zebrafish. Ilar J, 2012; 53(2):192–204; doi: 10.1093/ilar.53.2.192
Contreras-GuzmánES. Bioquímica De Pescado E Derivados. FUNEP: Jaboticabal; 1994.
19.
TsaiS, RawsonDM, ZhangT. Development of cryopreservation protocols for early stage zebrafish (Danio rerio) ovarian follicles using controlled slow cooling. Theriogenology, 2009 May;71(8):1226–1233; doi: 10.1016/j.theriogenology.2009.01.014
20.
SelmanK, WallaceRA, SarkaA, QiX. Stages of oocyte development in the zebrafish, Brachydanio rerio. J Morphol, 1993; 218(2):203–224; doi: 10.1002/jmor.1052180209
21.
Wilson-LeedyJG, IngermannRL. Development of a novel CASA system based on open source software for characterization of zebrafish sperm motility parameters. Theriogenology [Internet], 2007; 67(3):661–672; doi: 10.1016/j.theriogenology.2006.10.003
22.
HancockJL. The morphology of boar spermatozoa. J R Microsc Soc, 1957; 76(3):84–97; doi: 10.1111/j.1365-2818.1956.tb00443.x
23.
Streit JuniorDP, MoraesGV, RibeiroRP, PovhJA, SouzaED, OliveiraCAL. Avaliação de diferentes técnicas para coloração de sêmen de peixes. Arquivos De Ciências Veterinárias e Zoologia Da UNIPAR, 2004; 7:157–162. Available from: https://revistas.unipar.br/index.php/veterinaria/article/view/82/62
24.
SanchesEA, CaneppeleD, OkawaraRY, DamascenoDZ, BombardelliRA, RomagosaE. Inseminating dose and water volume applied to the artificial fertilization of Steindachneridion parahybae (Steindachner, 1877) (Siluriformes: Pimelodidae): Brazilian endangered fish. Neotrop Ichthyol, 2016; 14(1); doi: 10.1590/1982-0224-20140158
25.
MilioriniAB, MurgasLDS, RosaPV, OberlenderG, PereiraGJM, da CostaDV. A morphological classification proposal for curimba (Prochilodus lineatus) sperm damages after cryopreservation. Aquaculture Research, 2011; 42(2):177–187; doi: 10.1111/j.1365-2109.2010.02575.x
26.
GryshkovO, HofmannN, LauterboeckL, PogozhykhD, MuellerT, GlasmacherB. Multipotent stromal cells derived from common marmoset Callithrix jacchus within alginate 3D environment: Effect of cryopreservation procedures. Cryobiology [Internet], 2015; 71(1):103–111; doi: 10.1016/j.cryobiol.2015.05.001
27.
TsaiS, RawsonDM, ZhangT. Development of in vitro culture method for early-stage zebrafish (Danio rerio) ovarian follicles for use in cryopreservation studies. Theriogenology, 2010; 74(2):290–303; doi: 10.1016/j.theriogenology.2010.02.013
28.
MarquesLS, FossatiAA, LealMS, RodriguesRB, BombardelliRA, StreitDP. Viability assessment of primary growth oocytes following ovarian tissue vitrification of neotropical teleost pacu (Piaractus mesopotamicus). Cryobiology, 2018; 82:118–123; doi: 10.1016/j.cryobiol.2018.03.009
29.
GrelaE, KozłowskaJ, GrabowieckaA. Current methodology of MTT assay in bacteria—A review. Acta Histochem, 2018; 120(4):303–311; doi: 10.1016/j.acthis.2018.03.007
30.
WangG, KangN, GongH, et al.Upregulation of uncoupling protein Ucp2 through acute cold exposure increases post-thaw sperm quality in zebrafish. Cryobiology, 2015; 71(3):464–471; doi: 10.1016/j.cryobiol.2015.08.016
31.
YangH, DalyJ, CarmichaelC, MatthewsJ, VargaZM, TierschT. A procedure-spanning analysis of plasma membrane integrity for assessment of cell viability in sperm cryopreservation of Zebrafish Danio rerio. Zebrafish, 2016; 13(2):144–151; doi: 10.1089/zeb.2015.1176
32.
MahapatraBK, PailanGH, DattaS, SardarP, MunilkumarS. Chapter: Rigor mortis and fish spoilage. In: Manual on Fish Processing and Value Added Fish Products. Edition: 3rd. Central Institute of Fisheries Education: Mumbai, India. 2013. Available from: https://www.researchgate.net/publication/259357488_Rigor_mortis_and_fish_spoilage
33.
SavitriIKE, ApituleyYMTN, BawoleD, TuapetelF. Quality control of small pelagic fish stocks in distribution line in Ambon and Kei Kecil, Maluku. IOP Conf Ser: Earth Environ Sci, 2019; 339(1):12055; doi: 10.1088/1755-1315/339/1/012055
34.
MüllerK, MüllerP, PincemyG, KurzA, LabbeC. Characterization of sperm plasma membrane properties after cholesterol modification: Consequences for cryopreservation of rainbow trout spermatozoa. Biol Reprod, 2008; 78(3):390–399; doi: 10.1095/biolreprod.107.064253
CabritaE, Martínez-PáramoS, GavaiaPJ, et al.Factors enhancing fish sperm quality and emerging tools for sperm analysis. Aquaculture, 2014; 432:389–401; doi: 10.1016/j.aquaculture.2014.04.034
37.
Hosseinzadeh ColagarA, KarimiF, JorsaraeiSGA. Correlation of sperm parameters with semen lipid peroxidation and total antioxidants levels in Astheno- and Oligoasheno- teratospermic men. Iran Red Crescent Med J, 2013; 15(9):780–785; doi: 10.5812/ircmj.6409/
38.
HumphriesS, EvansJP, SimmonsLW. Sperm competition: Linking form to function. BMC Evol Biol, 2008; 8(1):319. Available from: https://doi.org/10.1186/1471-2148-8-319
39.
da CostaBB, PovhJA, SanchesEA, et al.Characterization of sperm quality in Brycon hilarii: How does morphology affect sperm movement? Theriogenology Wild [Internet], 2022; 1:100007; doi: 10.1016/j.therwi.2022.100007
