The Kitende-based side's 25-game unbeaten home run was for 19 wins and six draws in all competitions.
Before facing Simba SC in the CAF Champions League on Saturday, Vipers SC had gone 25 games without defeat at St Mary's Stadium, Kitende.
However, the Uganda Premier League champions could not extend that run to 26 games following a 1-0 loss to the Tanzanian giants last night.
The visitors' defender Henock Ngonga scored the game's only goal to break the hosts' unbeaten home run under the floodlight - a first at Kitende.
In Saturday's encounter, the Venoms had last tasted a home defeat in November 2021 when they lost 2-0 to URA FC in the league.
The Kitende-based side's 25-game unbeaten home run was for 19 wins and six draws in all competitions.
Vipers beat Tooro United 1-0, UPDF 7-2, Villa 3-0, Arua Hill 3-1, Bright Stars 2-1, Busoga United 2-0, Mbarara City 2-0, Gaddafi 2-0, Express 3-0, Police 5-0, Maroons 3-0, Arua Hill 1-0, UPDF 4-0, Villa 2-0, Onduparaka 4-0, and BUL 2-1 in the league.
The Venoms defeated Kataka 3-0, Mbarara City 4-0 in the Uganda Cup, and the Central African Republic Olympic Real de Bangui 1-0 in the Champions League.
Vipers' six draws were against KCCA 2-2, BUL 1-1, Bright Stars 0-0, Wakiso Giants 0-0 in the league and the Democratic Republic of Congo outfit TP Mazembe 0-0, Guinea side Horoya 0-0 in the Champions League.