The aim of this study was to compare the spermatozoa quality parameters in spermatozoa of RAS (Recirculating Aquaculture System; RAS group) cultured (commercial pellets) and natural condition cultured (WILD group) burbot Lota lota (live prey, Pseudorasbora parva). Seven of nine fish of the RAS group produced sperm, with sperm from only four of the fish having a motility of >5%. Sperm were collected from all nine fish of the WILD group, and sperm of six of the fish from the WILD group had motility of about 100% and three had sperm with 50% to 60% motility. Spermatozoa from the RAS group had a delay in activation compared to the WILD group. Fish from the RAS group also had a lesser volume of sperm (1.8 ± 1.2 mL) collected compared to the WILD group (3.6 ± 1.2 mL). Compared to the RAS group, sperm of the WILD group had a greater proportion of saturated fatty acids (SFA), as well as the phospholipid, phosphatidylethanolamine. The findings indicate that fish grown in natural conditions may be more suitable as broodstock. Ongoing research to develop methods of enhancing reproductive performance of burbot broodstock cultured in RAS is needed to investigate whether the quality of sperm can be improved by adjusting environmental conditions, diet, or combination of these factors.
- MeSH
- analýza spermatu * MeSH
- divoká zvířata MeSH
- Gadiformes metabolismus fyziologie MeSH
- lipidy analýza MeSH
- mastné kyseliny analýza metabolismus MeSH
- metabolismus lipidů * MeSH
- spermie chemie metabolismus MeSH
- vodní hospodářství metody MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
In most teleost fishes, sperm cells are quiescent in the seminal plasma and are activated by either a drop (fresh water fish) or an increase in osmolality (marine fish) when released in the water. It is most interesting to examine how the mechanisms of sperm motility activation can adapt to a broad range of salinities, as applies to some euryhaline species, and particularly to the tilapia Sarotherodon melanotheron heudelotii, which can reproduce at salinities from 0 up to 120 in the wild. Here, the gonado-somatic index, semen characteristics, and the osmotic and ionic requirements of sperm motility activation were compared in S. m. heudelotii reared in fresh water (FW), sea water (SW), or hypersaline water (HW; salinities of 0, 35, and 70, respectively). No salinity-dependent differences were found in gonado-somatic index or semen characteristics, except for an increase of seminal plasma osmolality with increasing salinity (from 318 to 349 mOsm kg(-1) in FW and HW fish, respectively). The osmolality range allowing the highest percentages of sperm activation broadened and shifted toward higher values with increasing fish ambient salinity (150-300, 300-800, and 500-1200 mOsm kg(-1), for FW, SW, and HW fish, respectively). Nevertheless, at the three fish rearing salinities, sperm could be activated in media that were hypotonic, isotonic, or hypertonic relative to the seminal plasma, at least when some calcium was present above a threshold concentration. The [Ca(2+)] required for the activation of S. m. heudelotii sperm is (1) higher in fish reared at a higher salinity (2) higher in hypertonic than that in hypotonic activation media, whatever the fish rearing salinity, and (3) higher in the presence of Na(+) or K(+), the negative effects of which increased with an increase in fish rearing salinity. The [Ca(2+)]/[Na(+)] ratios allowing for maximal sperm motility in SW or HW fish are close to those observed in natural environments, either in sea or hypersaline waters. In comparison to most teleosts with external fertilization, the total duration of sperm motility in S. m. heudelotii was exceptionally long (>2 hours regardless the fish rearing salinities). The decrease in sperm activity with increasing time since activation did not result from limiting energy reserves, as the addition of calcium in the activation medium caused most spermatozoa to become motile again. The comparison of sperm characteristics of S. m. heudelotii acclimated from FW to SW or HW with those of fish maintained all lifelong at their native salinity showed that adaptive responses were completed within 2 months or less.
- MeSH
- cichlidy fyziologie MeSH
- motilita spermií účinky léků MeSH
- osmotický tlak MeSH
- salinita * MeSH
- sperma fyziologie MeSH
- spermie fyziologie MeSH
- tolerance k soli MeSH
- viabilita buněk MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
This study examined the effect of cryoprotectants on DNA integrity, antioxidant defense, and resistance to oxidative stress in cryopreserved sterlet Acipenser ruthenus sperm. The freeze-thaw process significantly influenced sperm motility, with significant differences among cryoprotectants. In vitro exposure of cryopreserved sperm to the xanthine-xanthine oxidase (X-XO) system as a model reactive oxygen species inducer resulted in a lesser motility rate and velocity compared to the control, and there was a decrease in these variables in a time- and dose-dependent manner. The greatest X (0.6mM)-XO (0.05U/mL) concentration and incubation period (30min) was associated with 62% DNA fragmentation in sperm cryopreserved with 10% ethylene glycol (EG). The maximum lipid peroxidation (LPO) and carbonyl derivatives of proteins (CP) was also observed in sperm cryopreserved with 10% EG and exposed to the X-XO system at a concentration of 0.6mM X-0.05U/mL XO. The frozen/thawed sperm containing 10% EG and that with 10% dimethyl sulfoxide (DMSO) had a significant enhancement of superoxide dismutase (SOD) and glutathione reductase (GR) activity. The current study confirms that EG is not effective for cryopreservation, and sterlet sperm were highly sensitive to free radicals after cryopreservation with EG.
- MeSH
- acetamidy farmakologie MeSH
- dimethylsulfoxid farmakologie MeSH
- DNA účinky léků MeSH
- ethylenglykol farmakologie MeSH
- fragmentace DNA účinky léků MeSH
- kryoprezervace veterinární MeSH
- kryoprotektivní látky farmakologie MeSH
- methanol farmakologie MeSH
- oxidace-redukce účinky léků MeSH
- oxidační stres účinky léků MeSH
- ryby * fyziologie MeSH
- spermie účinky léků MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The nucleation and growth of crystalline ice during cooling, and further crystallization processes during re-warming are considered to be key processes determining the success of low temperature storage of biological objects, as used in medical, agricultural and nature conservation applications. To avoid these problems a method, termed vitrification, is being developed to inhibit ice formation by use of high concentration of cryoprotectants and ultra-rapid cooling, but this is only successful across a limited number of biological objects and in small volume applications. This study explores physical processes of ice crystal formation in a model cryoprotective solution used previously in trials on vitrification of complex biological systems, to improve our understanding of the process and identify limiting biophysical factors. Here we present results of neutron scattering experiments which show that even if ice crystal formation has been suppressed during quench cooling, the water molecules, mobilised during warming, can crystallise as detectable ice. The crystallisation happens right after melting of the glass phase formed during quench cooling, whilst the sample is still transiting deep cryogenic temperatures. We also observe strong water isotope effects on ice crystallisation processes in the cryoprotectant mixture. In the neutron scattering experiment with a fully protiated water component, we observe ready crystallisation occurring just after the glass melting transition. On the contrary with a fully deuteriated water component, the process of crystallisation is either completely or substantially supressed. This behaviour might be explained by nuclear quantum effects in water. The strong isotope effect, observed here, may play an important role in development of new cryopreservation strategies.
In the present study, for the first time in fish spermatozoa, we describe the precise chronology of motility initiation of sterlet (sturgeon) sperm from completely immotile flagella to regular full wave propagation. The successive activation steps were investigated by high-speed video microscopy, using specific experimental situation, where sperm motility initiation was delayed in time up to several seconds (10 ± 2.68 seconds). Starting from fully immotile, the flagellum shows some trembling for a brief period, soon followed by appearance of the first real bend (so-called "principal bend") with a large wave amplitude 4.28 ± 0.65 μm, then by the "reverse bend," the latter presenting a lower (P < 0.05) wave amplitude (1.14 ± 0.32 μm). This couple of first bends formed at the basal region begins to propagate toward the flagellar tip but gradually fades when reaching the midflagellum, wherein consequently the sperm cell remains nonprogressive. This behavior repeats several times until a stage where the amplitude of the reverse bend gradually reaches a value similar that of the principal bend: The larger amplitude of this couple of bends finally leads to sustain a real "takeoff" of the sperm cell characterized by a full flagellar wave propagation generating an active forward displacement similar to that occurring during regular steady state motility (several seconds after activation). Starting from the earliest stages of motility initiation, the wave propagation along the flagellum and formation of new waves proceeded in a helical manner leading to a 3-dimensional rotation of the whole spermatozoon. Eventually, we estimated that the time period needed from the activation signal (contact with fresh water) to full wave propagation ranges from 0.4 to 1.2 seconds.
- MeSH
- flagella fyziologie MeSH
- motilita spermií * MeSH
- ryby fyziologie MeSH
- spermie cytologie fyziologie MeSH
- vodní hospodářství MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cryopreservation causes osmotic changes and oxidative damage that have sublethal and lethal effects on spermatozoa. We examined these osmotic and oxidative effects on common carp spermatozoa motility; membrane integrity; levels of thiobarbituric-acid-reactive substance (TBARS) and carbonyl groups (CP); and the activity of superoxide dismutase (SOD), glutathione reductase, and glutathione peroxidase (GPx). Sperm was diluted in dimethyl sulfoxide (DMSO) and ethylene glycol-based extenders, followed by equilibration, freezing, and thawing. Equilibration in DMSO extender resulted in a significant reduction of spermatozoa motility, but motility was induced in those spermatozoa following dilution with saline buffer, which usually inhibits undiluted spermatozoa motility. Spermatozoa velocity and membrane integrity decreased with both extenders following freezing and thawing. No significant difference in levels of TBARS or CP, or in SOD activity, was seen in samples equilibrated with either extender. The freeze/thaw process induced significantly higher levels of TBARS, CP, and GPx activity, but did not affect the level of SOD. Glutathione reductase activity was inhibited in samples exposed to DMSO extender. Ethylene glycol should be considered a preferred cryoprotective agent for common carp spermatozoa to reduce osmotic and oxidative stress during cryopreservation.
- MeSH
- buněčná membrána účinky léků MeSH
- dimethylsulfoxid farmakologie MeSH
- ethylenglykol farmakologie MeSH
- glutathionperoxidasa metabolismus MeSH
- glutathionreduktasa metabolismus MeSH
- kapři metabolismus MeSH
- karbonylace proteinů účinky léků MeSH
- kryoprezervace metody veterinární MeSH
- kryoprotektivní látky farmakologie MeSH
- látky reagující s kyselinou thiobarbiturovou metabolismus MeSH
- motilita spermií účinky léků MeSH
- osmotický tlak účinky léků MeSH
- oxidační stres účinky léků MeSH
- rybí proteiny metabolismus MeSH
- spermie cytologie účinky léků metabolismus MeSH
- superoxiddismutasa metabolismus MeSH
- videomikroskopie veterinární MeSH
- vodní hospodářství metody MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
