Summary The objective of this work was to evaluate the sperm motility of 13 Steindachneridion parahybae males using open‐source software (ImageJ/CASA plugin). The sperm activation procedure and image capture were initiated after semen collection. Four experimental phases were defined from the videos captured of each male as follows: (i) standardization of a dialogue box generated by the CASA plugin within ImageJ; (ii) frame numbers used to perform the analysis; (iii) post‐activation motility between 10 and 20 s with analysis at each 1 s; and (iv) post‐activation motility between 10 and 50 s with analysis at each 10 s. The settings used in the CASA dialogue box were satisfactory, and the results were consistent. These analyses should be performed using 50 frames immediately after sperm activation because spermatozoa quickly lose their vigor. At 10 s post‐activation, 89.1% motile sperm was observed with 107.2 μm s−1 curvilinear velocity, 83.6 μm s−1 average path velocity, 77.1 μm s−1 straight line velocity; 91.6% were of straightness and 77.1% of wobble. The CASA plugin within ImageJ can be applied in sperm analysis of the study species by using the established settings.

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RESUMO -Um experimento foi realizado com o objetivo de estimar a concentração espermática das espécies dourado (Salminus brasiliensis), curimba (Prochilodus lineatus), jundiá (Rhamdia quelen), cascudo-preto (Rhinelepis aspera) e tilápia-do-nilo (Oreochromis niloticus) pelo método de espermatócrito. Utilizaram-se 19, 58, 51, 43 e 85 reprodutores de dourado, curimba, jundiá, cascudo-preto e tilápia-do-nilo, respectivamente. Com exceção da tilápia-do-nilo, os reprodutores foram submetidos ao processo de indução hormonal e posteriormente submetidos a coleta de sêmen. Foram comparadas as técnicas de mensuração da concentração espermática do sêmen por contagem em câmara hematimétrica de Neubauer e por espermatócrito. Os resultados obtidos foram submetidos à análise de regressão a 5% de probabilidade. As concentrações espermáticas mensuradas por ambas as técnicas apresentaram relação linear, para curimbas, jundiás e tilápias-do-nilo, com equações y = 6,6624 × 10 9 + 3,68553 × 10 8 x; y = 2,153 × 10 9 + 4,426 × 10 8 x e y = -9,0897 × 10 8 + 6,0167 × 10 8 , respectivamente. O método de espermatócrito pode ser utilizado para estimar a concentração espermática do sêmen de curimbas, jundiás e tilápias-do-nilo. Palavras-chave: Oreochromis niloticus, Prochilodus lineatus, reprodução de peixes, Rhamdia quelen, Rhinelepis aspera, Salminus brasiliensisSperm concentration estimate of fish semen using spermatocrit method ABSTRACT -The objective of this experiment was estimate sperm concentration of "dourado" (Salminus brasiliensis), "curimba" (Prochilodus lineatus), "jundiá" (Rhamdia quelen), "cascudo-preto" (Rhinelepis aspera) and Nile tilapia (Oreochromis niloticus) by the method of spermatocrit. It was used 19, 58, 51, 43 and 85 brood fish of "dourado", "curimba", "jundia", "cascudo-preto" and Nile tilapia, respectively. Except for the Nile tilapia, the fishes were subjected to the hormonal induction process and then submitted to the semen collection. The measurement techniques of the sperm concentration by sperm count in Neubauer chamber (CSPZ) and by spermatocrit (ESPMT) were compared. The results were submitted to the regression analysis at 5% of probability. It was observed that the variables CSPZ and ESPMT presented linear relationship to the "curimba", "jundia" and Nile tilapia with equations y = 6.6624 × 10 9 + 368553 × 10 8 x; y = 2.153 × 10 9 + 4.426 × 10 8 x and y = -9.0897 × 10 8 + 6.0167 × 10 8 , respectively. The method of spermatocrit can be used to estimate the sperm concentration of "curimba", "jundiá" and Nile tilapia.

This study describes the seminal and spermatic characteristics of fresh semen of Steindachneridion melanodermatum and investigates the effects of dilution, temperature, and storage period on its spermatic parameters. Sperm samples were collected from nine hormonally-induced males. The following parameters in fresh sperm were analyzed: seminal plasma osmolality (OSM), seminal pH, sperm motility (MOT), sperm velocity (SV) (including sperm curvilinear velocity (VCL), sperm straight-line velocity (VSL), and sperm average path velocity (VAP)), total time of sperm motility (TEMP), sperm concentration (CONC), and index of sperm normality (NORM). Sperm samples from each male were diluted in a solution containing 5% fructose and 5% powdered milk, and stored at 10°C and 25°C. The same was carried out for sperm samples not subjected to dilution. From these samples, MOT, VCL, VSL, VAP, SV, and TEMP were measured after 0 h, 5 h, 9 h, 18 h, 27 h, 36 h, 45 h, and 54 h. Males released 11.74 ± 5.38 mL of sperm, with an osmolality of 258.78 ± 29.36 mOsm.kg -1 and pH of 7.11 ± 0.31. The sperm presented a MOT of 99.86 ± 0.31% at a concentration of 1.03 × 10 10 ± 3.65 × 10 9 spermatozoa. The results indicate that sperm motility, sperm velocity, and total time of sperm activation were affected by dilution, storage temperature, and storage period (p < 0.05). Procedures for semen storage should be performed with undiluted sperm cooled at 10°C, or kept undiluted at 25°C for up to 27 h. Key words: CASA, endemic, sperm characterization, sperm motility, sperm storage, sperm velocity ResumoO objetivo deste estudo foi descrever os parâmetros seminais e espermáticos no sêmen fresco do Steindachneridion melanodermatum e os efeitos da diluição, da temperatura e do tempo de estocagem do sêmen sobre os parâmetros espermáticos. Foi coletado o sêmen de nove machos, induzidos hormonalmente. No sêmen fresco foram avaliados os parâmetros: osmolaridade do plasma seminal (OSM), pH seminal (pH), motilidade (MOT) e velocidades espermáticas (VCL, VLR, VMD, VE),