FC Bendigo's winning run was ended in Sunday's clash with Box Hill United in the National Premier League Victoria's division one match at Wembley Park.
The FC Bendigo line-up, captained by Trent Waterson, faced one of the league's best.
Box Hill United scored from a set-piece in the first half.
Although Bendigo had a lot more possession and attacking play in the second half, Box Hill broke through twice to achieve a 3-0 victory.
FC Bendigo's playing coach Steve Martin said Box Hill was a "well-drilled unit" which capitalised on errors.
"The final margin was disappointing, but our effort was excellent," Martin said of the clash with the second-placed United.
"Some lapses in concentration were costly. For more than 75 minutes we were right in the game."
Bendigo's best included goalkeeper Sebastian Mattei and Viktor Rodriguez in attack and midfield.
Box Hill won the under-20s match, 5-1.
FC Bendigo is at home this Saturday night to play Springvale White Eagles. The under-20s start at 5pm and will be followed by the main game at 7pm on the Tom Flood Sports Centre pitch.