Storage and transportation of Prochilodus lineatus (Characiformes) sperm prior to cryopreservation.

In this study we compared post-thaw quality of P. lineatus sperm frozen shortly after collection, with sperm frozen after dilution and transportation, and up to 6h from collection. From each sperm sample (n=10 males) five aliquots were taken. One aliquot was diluted in the freezing medium (1 sperm:8 glucose:1 methyl glycol) and frozen ∼20min after collection in the field (control), while the other four aliquots were transported to the laboratory where freezing took place 3 or 6h after collection. From the transported aliquots, two were diluted 1:4 in glucose solution before transportation (diluted samples), while the other two were kept undiluted until freezing (undiluted samples). Thus the five treatments were: control, undiluted-3h, diluted-3h, undiluted-6h and diluted-6h. Post-thaw sperm was evaluated for membrane integrity, motility rate and velocities (curvilinear=VCL; average path=VAP; straight line=VSL). Post-thaw membrane integrity did not differ among the five treatments (48-60% intact sperm). Sperm motility rate was similar (P>0.05) between control (64%) and undiluted samples (60-62%) and higher (P<0.05) than that in diluted samples (35-45%), regardless the time after collection when freezing took place. Velocities were higher in control and in undiluted-3h samples (VCL of 254-265µm/s, VAP of 219-244µm/s and VSL of 134-147µm/s) than in diluted samples or samples frozen 6h after collection. P. lineatus sperm can be transported/shipped to the laboratory without decreasing its suitability for cryopreservation. Sperm should be kept undiluted during storage and be frozen within 3h.

Osmolality and composition of the extender during the cold storage of Prochilodus lineatus (Characiformes: Prochilodontidae) sperm

This study evaluates the effects of extender osmolality and composition on the cooling of Prochilodus lineatus sperm. Sperm was diluted in six extenders: two compositions (powdered coconut water(tm) = ACP; Beltsville Thawing Solution(tm) = BTS) x three osmolalities (285, 325, and 365 mOsm/kg) plus an undiluted control, and stored at 6-8°C. Motility rate and velocities (curvilinear, straight line, and average path) were determined every other day. Osmolality did not affect the quality of cooled sperm, thus data were pooled. Motility was higher on d 0 compared to the other days and diluted samples (85-90%) yielded higher motility than control (75%). On d 2, motility was higher in BTS-diluted samples and control, but on d 4 and 6, control yielded the highest motility. Velocities decreased from d 0 to 6 in diluted samples, but not in control. On d 0, velocities were higher in BTS-diluted sperm, but, on d 2, 4, and 6, control yielded higher velocities despite of the large variation among males. Thus P. lineatus sperm should be stored in BTS or without dilution, for a maximum of two days at 6-8°C. Extender osmolality between 285 and 365 mOsm/kg does not affect sperm quality during cold storage...

Neste trabalho avaliou-se os efeitos da osmolalidade e da composição do diluidor no sêmen de Prochilodus lineatus, após o resfriamento. O sêmen foi diluído em seis diluidores: duas composições (água de coco em pó(r) = ACP; Beltsville Thawing Solution(r) = BTS) x três osmolalidades (285, 325 e 365 mOsm/kg) mais uma alíquota sem diluição como controle e armazenadas a 6-8°C. A taxa de motilidade e velocidades (curvilinear, retilinear e média de percurso) foram determinadas a cada dois dias. A osmolalidade não afetou a qualidade do sêmen resfriado, dessa forma foi feito um 'pool' desses dados. A motilidade foi maior no d 0 comparado aos outros dias e as amostras diluídas (85-90%) apresentaram as maiores motilidades do que o controle (75%). No d 2, a motilidade foi maior nas amostras diluídas em BTS e controle, mas nos d 4 e 6, o sêmen controle apresentou as maiores motilidades. As velocidades diminuíram do d 0 para o d 6 nas amostras diluídas, mas não no controle. No d 0, as velocidades foram maiores nas amostras diluídas em BTS, mas, nos d 2, 4 e 6, o controle apresentou as maiores velocidades apesar da grande variação entre os machos. Assim, o sêmen de P. lineatus deve ser resfriado em BTS ou sem diluição (controle), por no máximo dois dias a 6-8°C. A osmolalidade do diluidor entre 285 e 365 mOsm/kg não afeta a qualidade do sêmen durante o resfriamento...