Short-term storage and management of sperm in vitro is an easy and economical process in which suitable extenders can be utilized to extend the storage period and prevent sperm function impairment. Therefore, the current study aimed to evaluate the effect of suitable extenders during the short-term storage of sterlet sperm and determine their fertilizing capacity and hatching success. Three extenders containing a composition of 16, 20, and 24 mM NaCl, 1 mM KCl, 0.1 mM CaCl2, 10 mM Tris, pH 8.0 with osmolarity of 46, 55, and 62 mOsm/kg, were used to dilute the sperm of four sexually mature sterlet males (n = 4). Using a CASA system, the motility and velocity of undiluted and diluted sperm with extenders (E1 - E3) were assessed over 6 days at 0-2 °C. The short-term stored diluted sperm was then used in the fertilization and hatching assay, and undiluted fresh and stored sperm was used as a control. A two-way factorial analysis of variance (ANOVA) model confirmed significant effects on sperm motility, curvilinear velocity (VCL), and straight-line velocity (VSL) (P < 0.001), as well as their interaction with the extender. The model was decomposed into a one-way ANOVA to examine the impacts of extenders and storage time. With increasing storage periods, the sperm motility and velocity gradually decreased for diluted sperm with three extenders (E1-E3) but sharply decreased for undiluted sperm (Control). The motility of undiluted sperm was found 3.77 ± 4.09% at 4 days, whereas sperm diluted with extenders showed 57.57 ± 12.33% (E1), 64.34 ± 11.86% (E2), and 61.40 ± 12.41% (E3) motility at 6 days. This study explored extenders optimized with higher osmolarity (39-62 mOsm/kg) and lower K+ (1 mmol/L) as the most suitable medium for storing sterlet sperm for 6 days. After 6 days post storage, sperm diluted with extenders E1-E3 achieved a fertilization rate of 31.29 ± 14.2%, 31.66 ± 8.84%, and 30.67 ± 10.02%, respectively, and hatching success of 29.58 ± 13.4%, 30.50 ± 7.89%, and 27.95 ± 9.62%, respectively with freshly ovulated eggs.

Department of Biotechnology and Genetic Engineering Noakhali Science and Technology University Noakhali 3814 Bangladesh

Department of Oceanography Noakhali Science and Technology University Noakhali 3814 Bangladesh

Faculty of Fisheries and Protection of Waters South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses Research Institute of Fish Culture and Hydrobiology University of South Bohemia in České Budějovice Zátiší 728 2 Vodňany 38925 Czech Republic

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Alavi SMH, Gela D, Rodina M, Linhart O (2011) Roles of osmolality, calcium—potassium antagonist and calcium in activation and flagellar beating pattern of sturgeon sperm. Comp Biochem Physiol A-Mol Integr Physiol 160(2):166–174 PubMed DOI

Alavi SMH, Rodina M, Gela D, Linhart O (2012) Sperm biology and control of reproduction in sturgeon:(I) testicular development, sperm maturation and seminal plasma characteristics. Rev Fish Biol Fish 22:695–717 DOI

Alavi SMH, Cosson J, Bondarenko O, Linhart O (2019) Sperm motility in fishes:(III) diversity of regulatory signals from membrane to the axoneme. Theriogenology 136:143–165 PubMed DOI

Billard R, Cosson J, Perchec G, Linhart O (1995) Biology of sperm and artificial reproduction in carp. Aquaculture 129(1–4):95–112 DOI

Bobe J, Labbé C (2010) Egg and sperm quality in fish. Gen Comp Endocrinol 165(3):535–548 PubMed DOI

Bronzi BP, Rosenthal H, Gessner J (2011) Global sturgeon aquaculture production: an overview. J Appl Ichthyol 27:169–175 DOI

Cejko BI, Żarski D, Palińska-Żarska K, Słowińska M, Kowalski RK (2019) Artificial seminal plasma improves motility and fertilisation capacity of common carp DOI

Cheng Y, Xin M, Gela D, Rodina M, Tučková V, Kašpar V, Siddique MAM, Shelton WL, Linhart O (2020) Optimization of sterlet ( PubMed DOI

Cheng Y, Vechtova P, Fussy Z, Sterba J, Linhartová Z, Rodina M, Tučková V, Gela D, Samarin AM, Lebeda I, Xin M, Zhang S, Rahi D, Linhart O (2021) Changes in phenotypes and DNA methylation of in vitro aging sperm in common carp PubMed DOI PMC

Ciereszko A, Dietrich GJ, Dietrich MA, Nynca J, Kuźmiński H, Dobosz S, Grudniewska J (2010) Effects of pH on sperm motility in several Salmoniformes species: DOI

Contreras P, Dumorne K, Ulloa-Rodríguez P, Merino O, Figueroa E, Farias JG, Valdebenito I, Risopatron J (2020) Effects of short-term storage on sperm function in fish semen: a review. Rev Aquac 12(3):1373–1389 DOI

Cosson J (2004) The ionic and osmotic factors controlling motility of fish spermatozoa. Aquac Int 12(1):69–85 DOI

Cosson J, Linhart O (1996) Paddlefish (

Dettlaff TA, Ginsburg AS, Schmalhausen OI (1993) Development of Prelarvae. Sturgeon fishes. Springer, Berlin, Heidelberg, pp 155–195

Dzyuba B, Boryshpolets S, Shaliutina A, Rodina M, Yamaner G, Gela D, Dzyuba V, Linhart O (2012) Spermatozoa motility, cryoresistance, and fertilizing ability in sterlet DOI

Dzyuba B, Cosson J, Boryshpolets S, Bondarenko O, Dzyuba V, Prokopchuk G, Gazo I, Rodina M, Linhart O (2014) In vitro sperm maturation in sterlet. Acipenser Ruthenus Reprod Biol 14(2):160–163 PubMed DOI

Fatira E, Havelka M, Labbé C, Depince A, Iegorova V, Psenicka M, Saito T (2018) Application of interspecific Somatic Cell Nuclear Transfer (iSCNT) in sturgeons and an unexpectedly produced gynogenetic sterlet with homozygous quadruple haploid. Sci Rep 8(1):5997 PubMed DOI PMC

Gage MJ, Macfarlane CP, Yeates S, Ward RG, Searle JB, Parker GA (2004) Spermatozoal traits and sperm competition in Atlantic salmon: relative sperm velocity is the primary determinant of fertilization success. Curr Biol 14(1):44–47 PubMed

Gallego V, Cavalcante SS, Fujimoto RY, Carneiro PCF, Azevedo HC, Maria AN (2017) Fish sperm subpopulations: changes after cryopreservation process and relationship with fertilization success in tambaqui ( PubMed DOI

Gallis JL, Fedrigo E, Jatteau P, Bonpunt E, Billard R (1991) Siberian sturgeon, Acipenser baeri, spermatozoa: effects of dilution, pH, osmotic pressure, sodium and potassium ions on motility. In: Williot P (ed) Acipenser. Cemagref Publication, pp 143–151

Gela D, Rodina M, Linhart O (2008) Artificial reproduction of sturgeon (

Gosálvez J, López-Fernández C, Hermoso A, Fernández JL, Kjelland ME (2014) Sperm DNA fragmentation in zebrafish ( DOI

Herráez MP, Ausió J, Devaux A, González-Rojo S, Fernández-Díez C, Bony S, Saperas N, Robles V (2017) Paternal contribution to development: sperm genetic damage and repair in fish. Aquaculture 472:45–59 DOI

Hochleithner M, Gessner J (2012) The sturgeons and paddlefishes (Acipenseriformes) of the world–biology and aquaculture. Aqua Tech Publications, Germany, pp 1–248

Iegorova V, Psenicka M, Lebeda I, Rodina M, Saito T (2018) Polyspermy produces viable haploid/diploid mosaics in sturgeon. Biol Reprod 99(4):695–706 PubMed PMC

Linhart O, Cosson J, Mims SD, Shelton WL, Rodina M (2002) Effects of ions on the motility of fresh and demembranated paddlefish ( PubMed DOI

Linhart O, Cosson J, Mims SD, Rodina M, Gela D, Shelton WL (2003) Effects of ions on the motility of fresh and demembranate spermatozoa of common carp ( DOI

Linhart O, Rodina M, Gela D, Kocour M, Vandeputte M (2005) Spermatozoal competition in common carp ( PubMed DOI

Linhart O, Alavi SMH, Rodina M, Gela D, Cosson J (2008) Comparison of sperm velocity, motility and fertilizing ability between firstly and secondly activated spermatozoa of common carp ( DOI

Linhart O, Cheng Y, Xin M, Rodina M, Tučková V, Shelton WL, Kašpar V (2020) Standardization of egg activation and fertilization in sterlet (

Magnotti C, Cerqueira V, Lee-Estevez M, Farias JG, Valdebenito I, Figueroa E (2018) Cryopreservation and vitrification of fish semen: a review with special emphasis on marine species. Rev Aquac 10(1):15–25 DOI

Moorehead WR, Biggs HG (1974) 2-Amino-2-methyl-1-propanol as the alkalizing agent in an improved continuous-flow cresolphthalein complexone procedure for calcium in serum. Clin Chem 20(11):1458–1460 PubMed DOI

Park C, Chapman FA (2005) An extender solution for the short-term storage of sturgeon semen. N Am J Aqualcult 67(1):52–57 DOI

Sarosiek B, Dryl K, Kucharczyk D, Żarski D, Kowalski RK (2014) Motility parameters of perch spermatozoa ( DOI

Shaliutina A, Hulak M, Gazo I, Linhartova P, Linhart O (2013) Effect of short-term storage on quality parameters, DNA integrity, and oxidative stress in Russian ( PubMed DOI

Shazada NE, Alavi SMH, Siddique MAM, Cheng Y, Zhang S, Rodina M, Kocour M, Linhart O (2024a) Short-term storage of sperm in common carp from laboratory research to commercial production—a review. Rev Aquac 16(1):174–189 DOI

Shazada NE, Zhang S, Siddique MAM, Cheng Y, Rodina M, Alavi SMH, Linhart O (2024b) Optimizing extenders for short-term storage of Sterlet (

Siddique MAM, Butts IAE, Cosson J, Linhart O (2016) First report on facultative parthenogenetic activation of eggs in sterlet sturgeon, PubMed DOI

Tavakoli S, Luo Y, Regenstein JM, Daneshvar E, Bhatnagar A, Tan Y, Hong H (2021) Sturgeon, caviar, and caviar substitutes: from production, gastronomy, nutrition, and quality change to trade and commercial mimicry. Rev Fish Sci Aquac 29(4):753–768 DOI

Toth GP, Ciereszko A, Christ SA, Dabrowski K (1997) Objective analysis of sperm motility in the lake sturgeon, DOI

Trigo P, Merino O, Figueroa E, Valdebenito I, Sánchez R, Risopatrón J (2015) Effect of short-term semen storage in salmon ( PubMed DOI

Tuset VM, Dietrich GJ, Wojtczak M, Słowińska M, De Monserrat J, Ciereszko A (2008) Relationships between morphology, motility, and fertilization capacity in rainbow trout ( DOI

Viveiros ATM, Nascimento AF, Orfão LH, Isaú ZA (2010) Motility and fertility of the subtropical freshwater fish streaked prochilod ( PubMed DOI

Wayman WR (2003) From gamete collection to database development: development of a model cryopreserved germplasm respiratory for aquatic species with emphasis on Sturgeon. Ph.D. Dissertation, Louisiana State University, p 286

Xin MM, Tuckova V, Rodina M, Kholodny V, Dadras H, Boryshpolets S, Shaliutina-Kolesova A, Linhart O (2018) Effects of antifreeze proteins on cryopreserved sterlet ( PubMed DOI

Xin M, Cheng Y, Rodina M, Tučková V, Shelton WL, Linhart O (2020) Improving motility and fertilization capacity of low-quality sperm of sterlet PubMed DOI