Use your IGPU and test each pcie slot individually to see if you just have faulty pcie slots.
Checked this, everything seems okay here
1000w PSU is pretty borderline for mining with 6 card rig, to be honest (Always give yourself headroom with PSU. Efficiency is better and You can have a nice night sleep knowing that You aren't pushing Your PSU too hard)
Try GEN2 on Your PCI-E lanes.
That is very true!! and yes I tried Gen 2 a still problems with the 3rd GPU

I really don't know what else to try, I completed another fresh install of windows thinking it would be maybe a reg key that had got corrupt but yeah didn't work..
Debating to buy 3 more PCI risers and possibly RMA the motherboard for a new one or different one..
I've tried to reach out to Asrock but they haven't got back to me yet. Its so frustrating because this motherboard is meant to be a mining board, I wasn't expecting any of these problems...
I RMAd one of my H81 MOBOs on 20.09 (no POST) and I haven't heard anything about it from then... It seems that a lot of AsRock H81 Pro boards are faulty in some stupid way

Hope everything works out for You and You can get a new MOBO fast if this problem doesn't get fixed!
Thats crazy, im glad you managed to get that sorted!! And thanks, I've literally spent nearly 12 hours today to try resolve this, have never been so stressed in such a long time haha. I think I will literally just RMA the board at this rate. Praying that will solve the issue.
No way really, did you have the same issue as me? Or was it something slightly different, I am also debating to just purchase 3 new PCI risers to see if this would actually make any difference