Ayton's relegation to Division Three of the Scarborough Saturday League was confirmed despite a brave display in their 5-3 loss to Duchess.
Ian Jenkinson twice put Duchess ahead, only for Adrian Kwiatkowski and Chris Milburn to make it 2-2 at the break.
Duchess wrapped up the win with goals from JP Watson (2) and sub Ian Laing, Tyler Beck replying for Ayton.
Paul Oldroyd shone for Ayton with Luke Midgley the Duchess star man.
In Division One, Edgehill eased to a 7-0 home win against a weakened Seamer.
The Edgehill scorers were Danny Glendinning (2), Mikey Barker, Tyler Richardson, Jackson Jowett, keeper Liam Giles and a 20-yard volley from Dan Jones.
Itis Itis Rovers maintained their impressive finish to the third division season when beating Goal Sports 4-2, ending the promoted side's unbeaten home record.
Tom Thornton put Rovers ahead after good work from Neil Forsyth, then a Niall Gibb penalty and Thornton's second from a Dave Young assist made it three.
Tom Wilde and Niall Prentice brought it back to 3-2, but Forsyth lashed home a loose ball to seal Rovers' victory.
Thornton was named man of the match for the winning side.
Fishburn Park and Sherburn played out a thriller, which ended 3-3.
Sherburn moved two goals clear courtesy of a fine solo effort from Sam Rackham and a Paul Mills penalty.
Alex Wiggins pulled one back to keep Park in the game just before half-time.
Harry Walmsley looked to have put Sherburn on the path to the points with a 30-yard free-kick, but the understrength side faded in the closing stages and Park grabbed a point.
Wiggins tapped home his second and Theo Clarke's drive put Fishburn on terms.
Sixteen-year-old keeper Kieran Purvis impressed for Fishburn, as did winger Jack Kipling.
Sleights Reserves powered to an 11-0 win at Eastway, with Rob Ingham, Ben Watson and Spencer Hewison hitting a brace each.
Ryan Roe, Taylor Humble, Carl Harris, Mike Dent and player-boss Rob Winspear, in his final game in charge, also notched.
Hunmanby United Reserves beat Scalby Reserves 3-1.
Nick Turley slotted Scalby ahead after a great ball from Tommy Day, but United levelled before the break.
Hunmanby added two more goals, despite playing uphill, in the closing stages to seal the win.
Matty Atkinson scored a brace for Hunmanby, with Oliver Milner bagging the other.