Spermatozoon ultrastructure and sperm cryopreservation of the Brazilian dry season spawner fish pirapitinga,Brycon nattereri

The aims of this study were to describe the fresh spermatozoon ultrastructure using scanning and transmission electron microscopy and to improve the sperm cryopreservation methodology for the freshwater fish pirapitinga Brycon nattereri. Extenders (BTS^? and NaCl), straw volumes (0.5 and 4.0 mL), thawing temperatures (30 and 60 °C) and activating agents (0.29% NaCl and 1% NaHCO₃) were tested. Methylglycol was used as a cryoprotectant agent and sperm was frozen in nitrogen vapour (dry‐shipper). Post‐thawed sperm motility rate, motility quality (score 0=no movement; 5=rapidly swimming spermatozoa), duration of motility and spermatozoon morphology were evaluated. Fresh spermatozoon was 35.06 μm long, the head was ovoid (2.00 × 1.22 μm) with no acrosome, the midpiece was 2.15 μm long and the flagellum was 30.90 μm long with the typical 9+2 axoneme arrangement. Post‐thawed sperm motility rate (70–79% motile sperm), motility quality (score 3.1–3.7) and morphology (9.3–11.6% abnormal spermatozoa) were not affected by any of the parameters tested. The duration of sperm motility was longer when triggered in 1% NaHCO₃ (392–1031 s) compared with 0.29% NaCl (144–338 s). Brycon nattereri sperm cryopreserved under the conditions described above yields over 70% motility and should last long enough to fertilize oocytes, even after 2 years of freezing.     

Fatty acid influence on Prochilodus lineatus (Characiformes,Prochilodontidae) embryo cryopreservation parameters.

This study aimed to evaluate the vitellogenic transference and incorporation of long‐chain polyunsaturated fatty acids (LC‐PUFA) into the membranes of Prochilodus lineatus embryos, aiming to increase the permeability to cryoprotectants and resistance to electric fields. One hundred thirty broodstock of P. lineatus were fed with control (C) or fish oil‐supplemented diets (FO) for 12 months. The fatty acid (FA) profle was determined using gas chromatography. For the neutral fraction, the FO group had a decrease in monounsaturated fatty acids (MUFA) and an increase in n3PUFA and, n6PUFA. To test for cryoprotectant toxicity, embryos were exposed for 20 min to a cryoprotectant solution of 1,2‐Propanediol (Prop) at a concentration of 5 or 6 molar (M). For FO, a reduction in survival of 33.1% was observed in 5 M, and no survival was observed at 6 M. Embryo samples were exposed the six polarized electric fields (3.4–51.6 joules), and with 11.2 J of energy, the control group exhibited reduced survival in 98.3% of the fish, whereas the FO presented superior resistance, exhibiting a survival similar to that of the OJ up to 40.2 J. We conclude that FA were transferred between P. lineatus broodstock to the embryos, with an increase in LC‐PUFA resulting in lower survival rates in the cryoprotectant test in the FO group and a greater physical plasticity of FO embryos to electrical field tests.     

Improved solution for vitrification of Prochilodus lineatus embryos based on the reduction in risk factors: Toxicity,osmotic responses and ice‐nucleation

The objective of our work was to describe a low toxicity cryoprotectant solution that allowed vitreous solid formation. Embryos of Prochilodus lineatus were submitted to sensitivity evaluations of six internal cryoprotectants (dimethyl sulphoxide – Me₂SO, dimethyl acetamide – DMA, dimethyl formamide – DMF, methanol – MET, glycerol – GLY and 1,2‐propanediol – PROP) at concentrations of 1–6 M; and two external cryoprotectants (sucrose – SUC and glucose – GLU) at concentrations of 0.1–1 M for 20 min. The capacity of the cryoprotectant solutions to exchange heat with the medium and to form glassy solids was evaluated by immersing 10 μl of cryoprotectant in liquid nitrogen. The PROP had a high survival rate at all concentrations evaluated, and was the only substance that allowed a vitreous solid formation. Thus, it is concluded that the PROP‐6 M was the most adequate solution for embryonic vitrification processes, because heat exchange between the system (PROP 6 M/embryos/liquid nitrogen) was faster than for other cryoprotectants and combinations thereof; has low toxicity, promote high rates of dehydration in short periods, and reach the vitreous state, being a good candidate to be used in the tests of embryonic vitrification.     

The influence of increased water temperature on the duration of spermatogenesis in a neotropical fish,Astyanax altiparanae (Characiformes,Characidae)

Fish Physiology and Biochemistry - In view of the established climate change scenario and the consequent changes in global temperature, it is essential to study its effects on animal...     

Preliminary study on testicular germ cell isolation and transplantation in an endangered endemic species Brycon orbignyanus (Characiformes: Characidae)

Fish Physiology and Biochemistry - We aimed to develop a simplified protocol for transplantation of Brycon orbignyanus spermatogonial stem cells (SSCs) into Astyanax altiparanae testes. Brycon...