Doncaster put the disappointment of last week's South Yorkshire derby defeat behind them as they cruised to victory on their opening foray in the British and Irish Cup.
Lynn Howells' reward for paying the new competion its respect was a victory by four tries to one.
Hudson Tonga'uhia got the ball rolling with a try in the fifth minute, after the forwards had rumbled into the Scottish visitors' 22.
Ali Warnock
missed the conversion but set them on their way again midway through the half with a precise kick into touch that set up a five-metre lineout. This time it was Chris Planchant who peeled off the maul to touch down and Warnock found his range with the conversion.
Ayr came back strongly with a sustained spell of pressure, and they showed their willingness to attack by twice opting to kick the ball into touch instead of taking the three points from a penalty goal. The Knights punished the Scots by preventing them from crossing their line and headed into the break 12-0 ahead.
Warnock extended the lead with two penalties, before in the 60th minute, Tonga'uhia broke down the left and kicked ahead for Matt Williams who sprinted through and touched down, Warnock converting.
Ayr flickered into life five minutes later when fly-half Ross Currie side-stepped past a marker and fed replacement prop Steven Adair on his inside to score, Mark Stewart converting. Ayr were threatening again when Doncaster scored their fourth try, Warnock intercepting a pass inside his own half and galloping clear to touch down, his conversion taking the score to 32-7. Chris Briers had a late try disallowed for an earlier infringement.
Doncaster: Carter, Flockhart, Briers, Tonga'uhia, Williams, Warnock, D'Arcy; Davies, Lawrie, Cusack, Griffiths, Townson, Boyde, Cochrane, Planchant. Replacements: Corsar, Boden, Thiede, Kettle, Grainger, Hallam, McColl.
Ayr: Steele, Wilson, Stewart, Anderson, Taylor, Currie, Hunter, Mitchell, McArthur, Sykes, Sutherland, Kelly, Crossan, Dunlop, Tippet. Replacements: Nimmo, Adair, Reid, Francis, McFarlane, Manning, Fenwick.