Comparison between Tris-buffer and INRA-82 extenders on the quality of chilled rabbit spermatozoa

<p>The aim of the present study was to compare the effects of tris-buffer and INRA-82 extenders on the quality of cooled rabbit spermatozoa. Pooled semen samples were collected from eight New Zealand White rabbit bucks, divided into three groups and diluted 1:5 with three different semen extenders: INRA-82, tris-citrate glucose (TCG) and tris-citrate trehalose (TCT). Following dilution, the samples were stored at 5°C for up to 48 h. Sperm motility was recorded at 24, 36 and 48 h post-cooling. Furthermore, sperm viability, morphology, membrane integrity, acrosome status and DNA integrity were assessed at 24 h post-cooling. We found that progressive motility percentages were significantly higher (<em>P</em>&lt;0.05) in samples diluted in INRA-82 (61.00, 53.50, 44.00% at 24, 36 and 48 h post-cooling, respectively) than those diluted in either TCG (39.25, 32.00, 19.75%) or in TCT (47.25, 40.50, 29.00%). We also reported that sperm viability, percentage of normal spermatozoa, percentage of spermatozoa with intact acrosome and DNA integrity after 24 h cooling were significantly higher (<em>P</em>&lt;0.05) in INRA-82 diluted samples compared to TCG and TCT diluted ones. In summary, our results show that dilution of rabbit semen in INRA-82 improved sperm physiological parameters post-cooling compared to those diluted in TCG or TCT. Our findings also suggest that INRA-82 is a promising diluent that can be used effectively to maintain the viability of chilled rabbit semen.</p>

Time and Dose-Dependent Effects of Viscum Album Quercus on Rabbit Spermatozoa Motility and Viability in Vitro

The target of this study was to evaluate the effect of extract of the European mistletoe – Viscum album quercus L. on spermatozoa motility and viability in vitro. The CASA system was used to determine the spermatozoa motility parameters at different time intervals (0, 1, 2 and 3 h) and spermatozoa viability was determined in five different doses of Viscum album quercus L [10 (QA), 6.6 (QB), 3.3 (QC), 2.5 (QD) and 2 (QE) mg/ml]. Results in experimental groups detected a significant deterioration on rabbit spermatozoa after 1, 2 and 3 hours, compared to the control. The initial total spermatozoa motility showed increased value for all doses of Viscum album quercus in comparison to control. After in vitro culture a dose–dependent decrease (QA: reduction of 69.7 %, QB: reduction of 40.9 %) was found. For the progressive spermatozoa most significant decrease (86.8 % for QA vs. 48.5 % for QB) was detected compared to the control after 3 hours of culture. Spermatozoa viability (MTT test) was decreased in all experiment groups at the end of experiment, but the differences were not significant. Significant alterations of membrane integrity were found in groups with the highest Viscum album quercus concentration (QA, QB), but acrosome integrity showed no significant changes. Results suggest negative dose– and time–dependent effect of Viscum album quercus at higher doses on spermatozoa motility and viability parameters in vitro.

Quality assessment of cryopreserved New Zealand white rabbit spermatozoa in INRA-82 extender containing different cryoprotectants

<p>The present study aimed to evaluate the effect of supplementation of INRA-82 semen extender with different cryoprotectants (dimethyl sulphoxide; DMSO vs. dimethyl formamide; DMF) on the quality of white New Zealand rabbit buck spermatozoa. We also investigated the possible association between the synergistic action of DMSO and DMF and their relation with INRA-82 extender composition. Semen was collected and pooled from 8 adult rabbit bucks. Pooled semen samples were diluted 1:1 with INRA-82 extender supplemented with DMSO 8%, DMF 8% or a combination of DMSO 4% and DMF 4%. The diluted semen samples were cryopreserved in 0.25 plastic straws. After thawing, progressive motility, sperm viability, sperm abnormalities, membrane integrity, acrosome status, viability index and DNA integrity were evaluated. The results showed that dilution of rabbit buck semen in INRA-82 supplemented with DMSO and DMF (4% each) before freezing significantly (<em>P</em>&lt;0.05) improved sperm motility (42.00%), percentage of live spermatozoa (45.30%), proportions of spermatozoa with intact acrosome (59.75%) and percentage of spermatozoa with non-fragmented DNA (86.04%), compared to those diluted in INRA-82 supplemented either with DMSO 8% (+9, +10, +5 and +7 percentage points, respectively) or with DMF 8% alone (+18 +18, +12 and +9 percentage points, respectively). In conclusion, dilution of rabbit buck semen before freezing with INRA-82 extender supplemented with a combination of DMSO 4% and DMF 4% improved quality of frozen-thawed New Zealand White rabbit spermatozoa. Furthermore, our results also suggest that supplementation of INRA-82 with DMSO or with DMF alone at higher concentrations deteriorates the sperm quality.</p>