Strathdale's bid for an undefeated championship season was quashed by Shepparton United on Sunday.
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The Blues went into the final round game with the Bendigo Amateur Soccer League championship secured, but they needed at least a draw to remain undefeated.
A goal in the 67th minute from substitute Ardit Gavri was all Shepparton United needed to inflict the Blues' first defeat of the season.
Strathdale pushed hard, but its cause was made harder by the sending off of forward Lonain Burnett in the 82nd minute for a second yellow card offence.
The Blues and Shepparton United now turn their attention to the Super Cup which starts this Sunday.
The top eight teams qualify for the Cup, which is a knockout competition played over three weeks. Strathdale and Shepparton United could meet in the semi-finals.
Shepparton South warmed up for the Super Cup by putting 11 goals past Golden City.
Skipper Joel Aitken scored five times to take his season tally to 23.
In the only other division one game played, third-placed Moama-Echuca defeated bottom side Epsom 4-1. Two former Epsom players - Cooper Arkinstall and Mitch Langenbacher - scored two goals each for Moama after Caleb Sanders had put the Scorpions in front early.
In the women's championship, Strathdale slipped to fifth on the ladder after it lost to title winners Shepparton United 2-0.
United's star striker Sophie Papalia scored both goals to finish with 48 goals for the championship season.
Shepparton United was a perfect 18 wins from 18 matches.
La Trobe Uni outclassed Shepparton 8-0, with three of the eight goals coming via own goals. Zoe Zabilowicz scored a brace for La Trobe.
Shepparton South moved into fourth place on the back of 2-0 win over Golden City.
Britney Scrivens found the back of the net twice.
Spring Gully United was forced to work hard for its 2-1 victory against Tatura.
An own goal and a Paige Conder strike did the damage for the Reds.
Super Cup qualifying finals draw for this Sunday:
Division one (all games at 3pm) - Strathdale (1) v Colts United (8) at Epsom, Eaglehawk (2) v Golden City (7) at Truscott Reserve, Moama-Echuca (3) v Shepparton South (6) at Shadforth Park, Tatura (4) v Shepparton United (5) at Stanley Avenue.
Women's division (all games at 10.30am) - Shepparton United (1) v Golden City (8) at Stanley Avenue, Eaglehawk (2) v Spring Gully (7) at Truscott Reserve, Colts United (3) v La Trobe Uni (6) at Epsom, Shepparton South (4) v Strathdale (5) at Shadforth Park.
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