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To split them up, remove their current pen as allowed and set the pen you want one to go to, then tell your colonist to manually move that animal. Repeat until you get them in each pen, then make sure all pens are allowed so your handlers wont automatically move them around.
Just make two pens, and set one pen to accept only male and the other pen to accept only female horses. That will completely segregate the two genders and prevent any further pregnancies.
Another option would be to sterilise the horses trough the medical operations tab. This also prevents further pregnancies from occurring.
https://steamcommunity.com/sharedfiles/filedetails/?id=2558978295&searchtext=Animals+age+filter