Sperm contamination by urine in Senegalese sole (Solea senegalensis) and the use of extender solutions for short-term chilled storage
González-López, Wendy Ángela
Duncan, Neil J.
Methods are needed to manage the sperm of Senegalese sole (Solea senegalensis), which will enable the industry to use artificial fertilisation to reproduce hatchery raised sole and implement breeding programs. The present study aimed to (a) describe the male reproductive and urinary system, (b) describe the effects of urine contamination on sperm quality and (c) examine the use of extenders for short term chilled storage of sole sperm. Nine males were dissected to describe the male reproductive and urinary system. A total of 49 males were examined and 32 (65.3%) provided adequate sperm samples of the study. Initially the samples were described by appearance (colour, transparency and fluidity) and sub-samples analysed for sperm quality, urea concentration, osmolality, pH and protein concentration. Cell concentration and sperm quality parameters, percentage motility, curvilinear velocity (VCL) and average path velocity (VAP), were measured using ImageJ CASA. Control samples and samples diluted (1:3) in six different extender solutions (modified Leibovitz, Ringer, NAM, Sucrose, Stor Fish® and Marine Freeze®) were stored short-term (4 °C) and tested zero, three, six and 24 h after collection. The close proximity of the reproductive and the urinary systems, especially the sperm ducts being attached to the urinary bladder makes obtaining sperm without urine contamination appear difficult. All the samples appeared to be contaminated with urine. Samples that appeared to be contaminated with urine (yellow colour) had similar spermatozoa cell concentration and urea concentration as samples that appeared not to be contaminated with urine (whitish colour), although motility was significantly lower in yellow samples. Seminal plasma urea concentration was positively correlated with osmolality. Cluster analysis grouped samples with significantly higher sperm quality and pH and significantly lower urea concentration and osmolality to indicate that urine contamination negatively affected sperm quality by increasing osmolality and decreasing pH. Amongst the six extender solutions Leibovitz and Marine Freeze® preserved significantly higher percentage motility 24 h after collection. Ringer, NAM and Stor Fish® were intermediate and Sucrose was similar to control samples that significantly decreased motility 3 h after collection. Taken together all sole sperm samples probably had urine contamination, which is difficult or impossible to avoid especially if all the sperm available needs to be collected. The extenders, Leibovitz and Marine Freeze® were used to maintain sperm quality and mitigate the negative effects of urine contamination. The collection and short term chilled storage in extenders of sole sperm from the majority of males in a broodstock (65.3%) can provide a valid sperm management system for industrial application for artificial fertilisation, however, further work is needed.
639 - Caça. Pesca. Piscicultura
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González-López, Wendy Ángela, Sandra Ramos-Júdez, Ignacio Giménez, and Neil J. Duncan. 2020. "Sperm Contamination By Urine In Senegalese Sole (Solea Senegalensis) And The Use Of Extender Solutions For Short-Term Chilled Storage". Aquaculture 516: 734649. Elsevier BV. doi:10.1016/j.aquaculture.2019.734649.
Grant agreement number
INIA-FEDER/Programa Estatal de I+D+I orientada a los retos de la sociedad/RTA2014-00048/ES/Gestión de reproductores basada en su comportamiento para aumentar la producción de gametos y el éxito reproductivo en el lenguado senegalés (Solea senegalensis) cultivado (F1) y salvaje/
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