In all experiments, semen was assessed for plasma membrane integrity and for percentage of motile and progressive sperm by CASA.
There were two different experiments, semen from six mature Holstein bulls (Bos Taurus) was frozen with different concentrations of OPN (0, 1, 10, 100 μg/mL).
Even though PRO 2000 appeared effective in in vitro experiments, semen reduced its efficacy in animal model testing which may explain the lack of protection in the clinical trials.
For the initial experiments semen samples were also obtained from consenting healthy donors involved in Spanish sperm donation programs and patients undergoing fertility treatment in another Fertility Clinic (Tambre Clinic, Madrid, Spain).
For experiment 2, semen from nine Belclare rams was evaluated by IVF and semen from rams with the highest (n = 3) and lowest (n = 2) IVF scores was used for cervical insemination of ewes (n = 111) under experimental conditions.
In Experiment 1, semen was diluted in Triladyl® extender and frozen with freeze rate −10 °C/min.
In Experiment 1, semen from the fast and slow treatments was used to fertilize ovine oocytes that had been matured in vitro.
In Experiment 2, semen was diluted in Triladyl® or TCA extender, and frozen with three different freeze rates, i.e. −10, −25 or −40 °C/min.
In experiment 2, semen stored at a constant 5°C resulted in a lower 60-d NRR (62.5%) than storage at constant 15°C or unregulated temperature or frozen-thawed semen (73.6, 74.6, and 74.4%, respectively.
The Macho Mojito was drinkable enough for me to want to experiment with semen a bit further.
