ABSTRACT The use of carp pituitary extract (CPE) in fish reproduction may have drawbacks, whereas analogues of mammalian (aGnRHm) or salmon (aGnRHs) gonadotropins containing dopamine inhibitor (DI) may offer greater advantages. The objective of this study was to evaluate the effect of different hormone inducers on the reproduction of Prochilodus brevis males. A total of 44 specimens of P. brevis were divided into six groups, namely, CPE 4.0 mg; saline solution 1.0 mL; aGnRHm + DI 0.3 mg and 0.5 mg; and aGnRHs + DI 0.25 mL and 0.35 mL kg-1 of live weight. The number of ejaculates, seminal volume, curvilinear velocity (VCL), straight-line velocity (VSL), average path velocity (VAP), sperm morphology and concentration, and membrane and DNA integrity were determined. Sample normality was examined using the Shapiro-Wilk test, and homogeneity using the Levene test. Analysis of variance (ANOVA) was conducted, followed by Tukey's test to compare means, with statistical significance set at p < 0.05. Notably, all fish induced with CPE 4.0 and aGnRHm + DI demonstrated ejaculation, yielding a greater seminal volume. Sperm concentration under aGnRHs + DI 0.35 exceeded that with aGnRHm + DI 0.3 and CPE 4.0. Regarding velocity, aGnRHm + DI 0.3 outperformed aGnRHs + DI 0.35 in VCL, while aGnRHm + DI 0.3 and 0.5 and aGnRHs + DI 0.25 led to higher VSL than aGnRHs 0.35. Additionally, VAP was elevated under aGnRHm + DI 0.3 and CPE, in contrast to aGnRHs + DI 0.35. Improved sperm morphology was observed with aGnRHm + DI 0.5 and aGnRHs + DI 0.25 compared to aGnRHm + DI 0.3. The remaining parameters did not differ significantly. (CPE drawbacks (aGnRHm (aGnRHs (DI advantages males 4 P groups namely 40 0 4. 10 1 1. 03 3 0. 05 5 025 25 0.2 035 35 kg1 kg kg- weight ejaculates volume VCL , (VCL) straightline straight line VSL, (VSL) VAP, (VAP) determined ShapiroWilk Shapiro Wilk ANOVA (ANOVA conducted Tukeys Tukey s means 005 0.05 Notably ejaculation Additionally significantly 02 2 (VCL (VSL (VAP 00 0.0

Abstract The cooling of Litopenaeus vannamei shrimp spermatophores for assisted insemination can enable the transfer of gametes between reproduction laboratories. This study aimed to assess three extenders for cooling L. vannamei spermatophores for assisted insemination. Spermatophores were chilled at 15 °C for 24 or 48 hours using powdered coconut water ACP® (PCW), mineral oil (MO), and sterilized seawater (SSW) as extenders. All treatments demonstrated consistent responses over time. Apparent viability and morphological integrity percentages remained above 60% and 70%, respectively, across treatments and storage durations. Focusing on diluents, normal cell percentages for MO, SSW, and PCW treatments were 74.9±9.20%, 77.3±9.40%, and 78.1±6.35%, respectively, irrespective of storage time. The highest hatching rate was observed in the SSW treatment (80.67±12.01%), which was significantly superior to the PCW treatment (50.15±20.75%). The hatching rates observed in the MO treatment (71.47±18.83%) did not statistically differ from either PCW or SSW treatments. The cooling protocol successfully preserved the spermatophores' ability to maintain favorable levels of apparent viability, normal morphology, and hatching rates after 48 hours of storage at 15 °C using mineral oil, seawater, or ACP® as extenders. Sterilized seawater emerged as the most efficient diluent, delivering superior hatching rates following artificial insemination. laboratories L 1 C 2 4 ACP PCW, , (PCW) (MO) (SSW time 60 70 70% respectively durations diluents 749920 74 9 20 74.9±9.20% 773940 77 3 40 77.3±9.40% 781635 78 6 35 78.1±6.35% 80.67±12.01%, 80671201 80.67±12.01% 80 67 12 01 (80.67±12.01%) 50.15±20.75%. 50152075 50.15±20.75% . 50 75 (50.15±20.75%) 71.47±18.83% 71471883 71 47 18 83 (71.47±18.83% morphology diluent (PCW (MO 7 74992 74.9±9.20 77394 77.3±9.40 78163 78.1±6.35 8067120 80.67±12.01 8 0 (80.67±12.01% 5015207 50.15±20.75 5 (50.15±20.75% 71.47±18.83 7147188 (71.47±18.83 7499 74.9±9.2 7739 77.3±9.4 7816 78.1±6.3 806712 80.67±12.0 (80.67±12.01 501520 50.15±20.7 (50.15±20.75 71.47±18.8 714718 (71.47±18.8 749 74.9±9. 773 77.3±9. 781 78.1±6. 80671 80.67±12. (80.67±12.0 50152 50.15±20. (50.15±20.7 71.47±18. 71471 (71.47±18. 74.9±9 77.3±9 78.1±6 8067 80.67±12 (80.67±12. 5015 50.15±20 (50.15±20. 71.47±18 7147 (71.47±18 74.9± 77.3± 78.1± 806 80.67±1 (80.67±12 501 50.15±2 (50.15±20 71.47±1 714 (71.47±1 74.9 77.3 78.1 80.67± (80.67±1 50.15± (50.15±2 71.47± (71.47± 74. 77. 78. 80.67 (80.67± 50.15 (50.15± 71.47 (71.47 80.6 (80.67 50.1 (50.15 71.4 (71.4 80. (80.6 50. (50.1 71. (71. (80. (50. (71 (80 (50 (7 (8 (5 (

Abstract The action of substances with non-permeable cryoprotectant potential, besides glucose, has not yet been studied for the species Prochilodus brevis. The objective of this work was to evaluate the action of four non-permeable cryoprotectants on this species sperm cryopreservation. Five pools were cryopreserved in a solution of 5% glucose and 10% dimethyl sulfoxide (Me2SO) associated or not (control) with cryoprotectants egg yolk (5, 10 or 12%), soy lecithin (2.5, 7.5 or 10%), sucrose (5, 10 or 20%) and lactose (5, 8 or 15%). After thawing, samples were evaluated for sperm kinetics (total motility, motility duration, velocities, and wobble - WOB), morphology and membrane and DNA integrity. The treatments containing egg yolk improved significantly (P<0.05) results when compared the control for the membrane integrity parameter. When compared to other treatments, egg yolk, at any concentration, presented higher results (P<0.05) for membrane integrity, total motility, curvilinear velocity (VCL) and average path velocity (VAP) parameters. Egg yolk also showed the best results for WOB, but it did not differ from 5% and 8% lactose and 5% and 20% sucrose. Soy lecithin had the lowest percentages of morphologically normal sperm (P<0.05), while the other treatments did not differ from each other. There was no difference regarding DNA integrity data. Thus, 5% egg yolk is indicated as a non-permeable cryoprotectant for P. brevis, in association with 5% glucose and 10% Me2SO.