Influence of different anaesthetic protocols over the sperm quality on the fresh, chilled (4°C) and frozen‐thawed epididymal sperm samples in domestic dogs

Abstract

This study assessed the influence of three different anaesthetic protocols on semen quality obtained from the epididymis. Sixty male dogs undergoing to routine sterilization were assigned to three anaesthetic protocols: thiopental group (TG, n = 20), propofol group (PG, n = 20) and ketamine-dexmedetomidine group (KDG, n = 20). Immediately after orchidectomy, the cauda epididymides and vas deferent ducts were isolated and then a retrograde flushing was performed to collect spermatozoa. In experiment 1, after the initial evaluation of the semen (sperm concentration, sperm motility and the percentages of live spermatozoa, abnormal spermatozoa and acrosome membrane integrity), semen samples were diluted in Tris-glucose-egg yolk extender and chilled for 48 hr, and the sperm motility was assessed at 6, 24 and 48 hr. In experiment 2, semen samples were diluted in Tris-glucose-egg yolk extender and chilled for 24 hr, and then samples were frozen in two extenders with different glycerol concentrations, to reach a final concentration of 50-100 × 10(6) spermatozoa ml(-1) , 20% egg yolk, 0.5% Equex and 4% and 5% glycerol, respectively. Mean values of total sperm concentration, sperm viability and the percentages of intact acrosome and abnormal spermatozoa were not significantly different between experimental groups, and therefore, the anaesthetic protocols assessed did not affect sperm parameters mentioned above. However, our study confirmed a detrimental effect of the use of thiopental (TG) over the total sperm motility (p < 0.05) and progressive sperm motility (p < 0.05) of the fresh and chilled epididymal sperm samples. The anaesthetic protocols including the application of propofol or ketamine-dexmedetomidine can be used to recover sperm in domestic canids without significant changes in sperm quality compared when semen is collected routinely and these techniques could be applicable to endangered wild canids.

Cloning and characterization of the rat Slo3 (KCa5.1) channel: From biophysics to pharmacology

“…In recent years, anaesthetic protocols used during reproduction assistance have attracted more concern regarding the quality of sperms (Batista et al, 2016; Vlahos et al, 2009). The inhibition of the Slo3 channel by anaesthetics has not been examined previously.…”

“…However, other mammalian orthologues of the Slo3 channel have not been cloned and characterized, despite their importance in the identification of the evolutionary route of Slo3 channels. In the meantime, as analgesia and anaesthesia are usually required for assisted reproductive technology, the effects of general anaesthetic on sperm quality have also been studied (Batista, Vilar, Rosario, & Terradas, 2016; He et al, 2016; Vlahos, Giannakikou, Vlachos, & Vitoratos, 2009). But whether general anaesthetics alter sperm quality through the Slo3 channel has not been addressed.…”

The effects of propofol and midazolam on rat spermatogenic parameters

“…Their effects on motility and density parameters have been currently pointed out in recent studies. In a study by Batista et al (2016) on dogs, thiopental, propofol and ketamine were used, and thiopental was reported to have a negative effect on motility rate. Our results were similar to this study.…”

Efficiency and cell viability implications using tip type electroporation in zebrafish sperm cells

“…The animals were fed ad libitum with commercial fish feed, twice daily (Tetra ColorBits). For each replicate of the experiment, 4 males with 4-6 months of age were sacrificed by sectioning the spinal cord (the accepted method when the use of anaesthesia can affect the results of sperm analysis) [23]. Through an abdominal incision and dissection, the gonads were removed and placed in a microtube containing 200 μL of Beltsville Thawing Solution-BTS at 4 ºC with pH 7.4 and an osmolality of 350 mOsm [24] obtaining a sperm pool from the different males.…”