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Dominguez-Rebolledo A.E.,National Wildlife Research Institute IREC | Martinez-Pastor F.,University of Leon | Bisbal A.F.,National Wildlife Research Institute IREC | Ros-Santaella J.L.,National Wildlife Research Institute IREC | And 6 more authors.
Reproduction in Domestic Animals | Year: 2011

Contents: Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H2O2) levels on red deer spermatozoa after cryopreservation, and the role of male-to-male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200μm H2O2 for 2h at 37°C. Intracellular reactive oxygen species (H2DCFDA-CM) increased with H2O2 concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200μm. Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50μm H2O2 and above. In a second experiment, samples from seven males were submitted to 0 and 200μm H2O2 for 2h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H2O2 presence. We found that the kinematic parameters reflected male-to-male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H2O2 or after incubation with H2O2. Red deer spermatozoa are relatively resilient to H2O2 after thawing, but it seems to be a great male-to-male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols. © 2010 Blackwell Verlag GmbH. Source


Maroto-Morales A.,National Wildlife Research Institute IREC | Berlinguer F.,University of Sassari | Fernandez-Santos M.R.,National Wildlife Research Institute IREC | Fernandez-Santos M.R.,Institute of Regional Development IDR | And 7 more authors.
Theriogenology | Year: 2011

The aim of this work was to study the effect of storage temperature during the transport of ovaries on cleavage and blastocyst rates in Iberian red deer, because wild populations of this subspecies are usually far from laboratories. A total of 472 ovaries from 236 Iberian hinds were recovered and maintained in saline solution at 5-8 °C or 20-25 °C for 12 h. After storage, aspirated oocytes were matured with FSH/LH or EGF and the developed embryos were cultured with oviduct epithelial cells monolayer (OCM). A higher (P = 0.009) cleavage rate was obtained when the ovaries were stored at 5-8 °C. However, there were no differences between both storage temperatures in relation to the percentage of blastocysts obtained. Considering the management and production systems of Iberian red deer, this study provides important information about the ovary storage temperature during transport with the purpose of assuring an optimal in vitro embryo production. © 2011 Elsevier Inc. Source

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