Staining methods suitable for spermatozoa morphology, morphometry and membrane integrity evaluation on sterlet (Acipenser ruthenus)

Igna, Violeta; Mihailov, Sandra; Grozea, A.

doi:10.1016/j.jbiotec.2018.06.102

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“…Differences in human spermatozoa head size values (length and width) were observed when two different staining methods were used: Papanicolau and Diff-Quik [ 45 ]. Our preliminary study was conducted to identify the optimal staining variant for Russian sturgeon spermatozoa and started with the same issues encountered in sterlet ( Acipenser ruthenus ) spermatozoa [ 46 ]. In our study, Spermac and Trypan Blue were found to be suitable for staining Russian sturgeon spermatozoa, providing an optimal visualization for morphological and morphometric analysis in bright field microscopy.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Russian sturgeon (Acipenser gueldenstaedtii) Semen Quality and Semen Cryopreservation

Igna

Telea²,

Florea³

et al. 2022

Animals

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The alarming decline in sturgeon populations doubled by growing demands for sturgeon products, urge us to prevent the loss of these species. Fish stocking in natural habitats and developing fish farms are viable solutions, which can be successfully implemented with the help of reproductive biotechnologies. Despite the fact that semen cryopreservation is admittedly an important step for saving the Russian sturgeon, a reproducible standard method with good results has yet to be identified. Sperm quality assessment is essential for quantifying the impact of cryopreservation on spermatozoa. The purpose of our study was to provide additional information regarding semen cryopreservation and semen quality evaluation for the Russian sturgeon. Our study method is based on the use of two yolk-free extenders (with different cryoprotectants: DMSO and methanol) for freezing semen, using a simple freezing protocol. Parameters such as volume, concentration, motility, morphology and membrane integrity were evaluated. In conclusion, cryopreservation of Russian sturgeon spermatozoa using an extender containing methanol as cryoprotectant led to high egg fertilization percentages (72.67 ± 5.4%) even if the total motility values recorded for thawed semen were quite low (18–25%). Additionally, we identified two optimal stains for morphological studies and morphometry (Spermac stain kit and Trypan Blue Solution).

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Section: Discussionmentioning

confidence: 99%