STOW-on-the-Wold Rugby Club went down 13-9 at lowly Alchester in Southern Counties North.
The sixth-placed Cotswold side were unable to follow up their fine 31-20 home success over high-flying Thatcham.
Chipping Norton, who are fifth in the table, secured another Berks/Bucks and Oxon Premier Division success, beating Gosford All Blacks 19-13.
Shipston-on-Stour continued their improvement with a 26-25 victory at Southam building on a fine 40-18 beating of Upton. They have moved up to third-from-bottom in Midlands Three West (South).
In football, Moreton Rangers kept up the pressure on Gloucestershire Northern Senior League Division Two leaders Barometrics with a 5-0 win at Viney St Swithins and host Leonard Stanley on Saturday (3pm).
Winchcombe Town, who entertain Viney on Saturday (3pm), lost 2-0 at Newent Town.