Sam Olver kicked two penalties, but the Wanderers were beaten 12-6 by Gloucester at Franklin’s Gardens on Monday night.
Three days after the Saints first team claimed a 15-3 Aviva Premiership win against the Cherry and Whites, the second string suffered a defeat to the club from Kingsholm.
Fly-half Billy Burns booted four penalties to secure the Gloucester success, with Olver’s double not enough for the Wanderers, who suffered their first Aviva A League loss of the season.
The home side had fielded a strong 15, with Jamie Elliott lining up at centre and Ken Pisi, Tom Stephenson, Tom Collins and Gareth Denman among the others involved.
But it was Gloucester who got off to the better start as Burns put them ahead from the tee, eight minutes after Olver had missed for the Wanderers.
The away side were playing with the wind at their backs, making kicking far easier for Burns than for Olver.
The Wanderers 10 missed a second penalty on 33 minutes and, again, Burns followed that by landing one for Gloucester, giving them a 6-0 half-time lead.
Gloucester suffered a sin-binning five minutes into the second period, with Elliott Stooke yellow carded.
Olver cut the gap with a penalty before Burns restored his team’s six-point advantage.
Steve McColl then missed an ambitious drop-goal attempt for Gloucester, but they had another three points when Burns again hit the target 15 minutes from time.
Olver’s 75th-minute penalty set up a tense finale, during which Burns was sin-binned, but Gloucester held on to claim the spoils.
Wanderers: Mallinder; K Pisi (Packman 61), Elliott (Eves 61), Stephenson, Collins; Olver, Kessell (Mitchell 66); Hobbs-Awoyemi, Williams (Dawes 38), Denman (Parkin 61), Onojaife (Moon 66), Woolford (capt), Marshall, Allman (Cardall 68), Bennett.