Alternating from bitch to bitch the left or right oviduct and uterine horn were flushed separately using 5 ml Tyrode medium per flushing.
After initial aspiration, follicles were flushed and aspirated three times each with 2 mL flushing medium with heparin (SynVitro® Flush, Origio, Berlin, Germany).
Batches of the culture in 120-mL serum bottles, each containing 40 mL of medium, were flushed with mixture gas of N2/CO2 (80 20, v/v) for 5 min and sealed with butyl rubber septums and aluminum crimp seals.
Batches of the culture in 160-mL pressure-resistant serum bottles, each containing 20 mL of medium, were flushed with N2 for 5 min, sealed with butyl rubber septums and aluminum crimp seals, and autoclaved at 121 °C for 20 min.
The medium was flushed with CO2 to maintain anaerobiosis.
Bottles with phosphate-buffered medium were flushed with N2 (100%), while bottles with bicarbonate-buffered medium were equilibrated with a mixture of N2 CO2 (80 20%).
For the growth experiments, serum bottles containing the culture medium were flushed with N2 CO2 (80/20) to remove any trace of oxygen in the bottles, capped with thick butyl-rubber stoppers, and autoclaved.
Subsequently the medium was flushed with a mixture of nitrogen and CO2 (80 : 20 v/v) for 5 minutes and inoculated with 4 mL inoculum (10% v/v inoculum), before being incubated at 37°C with continuous stirring (150 rpm).
Twenty-four embryos were recovered 7 d post-ovulation; all were subjected in sequential order to five washes in embryo flush medium, two trypsin treatments, and five additional washes in embryo flush medium (prior to transfer).
Non surgical flushing by cervical via was efficient to recover flushing medium and embryos in goats.
