BRISTOL City outclassed a tepid Saints to end their 25-match unbeaten streak with a comfortable 3-1 win at Ashton Gate.
Russell Martin's Saints lost ground on Championship promotion rivals Leeds United, who produced a resounding win of their own at Swansea City.
Sam Bell and Rob Dickie scored second-half goals for the hosts, who were deserving winners for a competent 90-minute performance, before a Harry Cornick third.
Adam Armstrong stepped up to convert a spot-kick in seven minutes of added time but it was merely consolation for the visitors.
Saints were looking to make it 23 games without defeat in league competition, which would have drawn them one shy of the club's all-time record.
Martin made four changes to his Saints team from Saturday's chaotic 5-3 win over Huddersfield, the first time they had come from two behind at half-time to win since April 2021.
There was a first league start for Joe Rothwell and first Saints start for David Brooks, with Shea Charles and Sam Edozie also recalled.
Robins coach Liam Manning also made four changes to his team that won 2-1 at Middlesbrough last time out, recording their first league victory of 2024.
Bristol City were managing tired legs after going to extra time and penalties with Premier League Nottingham Forest in the FA Cup fourth round.
City started much the brighter team but spurned their chances after doing Saints in transition over and over again.
Edozie went closest for Saints in the first period, looping a snapshot off the crossbar after a one-touch passage with Rothwell into the box.
Ashton Gate celebrated as Bell hit the side netting in a succession of chances which saw Gavin Bazunu come halfway to the centre circle to intercept a one-on-one.
The Irishman was the culprit himself for the final chance of the half, with his pass cut out but Jason Knight's penalty appeals under pressure from Kyle Walker-Peters were waved away.
Martin opted for a halftime change and Stuart Armstrong replaced the passive Charles, who struggled to fully get to grips with the contest.
There was an almost an immediate reaction as Brooks attacked a low ball in by Edozie - but the Welshman required treatment for slamming into the post.
Instead, it was the hosts who would take the lead. Bell tapped in from close range after a low ball had been played across the six-yard box from the Saints left.
Walker-Peters had a volley well saved but the Robins struck the second too, with Dickie rising highest to head in a Joe Williams corner.
A unanimous cry of 'Who are you' went up from the bouncing Ashton Gate, after the initial celebrations subsided.
Substitute Cornick added the third emphatically finishing from a central area following a rapid counter-attack and lethal ball into Bazunu's box.
Saints were unable to even look like avoiding defeat - let alone win a fifth consecutive league fixture for the first time since 2011.
Bristol City were also the victors over 29 unbeaten Leeds United back in 1974, winning 1-0 at Elland Road.
Armstrong did convert a consolation penalty for his 16th league goal of the season, beating O'Leary in seven minutes of added time after Walker-Peters was fouled.
Bristol City: O'Leary; James, Williams, Mehmeti (Mebude, 85), Knight (Garnder-Hickman, 64), Conway (Wells, 76), Dickie, Tanner, Bell (Cornick, 64), Roberts (McCrorie, 76), Vyner.
Unused subs: Bajic, McCrorie, King, Sykes, Wells, Knight-Lebel.
Booked: Vyner.
Saints: Bazunu; Walker-Peters, Harwood-Bellis, Bednarek, Manning; Charles (S Armstrong, h-t), Smallbone, Rothwell (Kamaldeen, 74), Brooks (Mara, 63), Edozie (Fraser, 63), A Armstrong.
Unused subs: Lumley, Stephens, Amo-Ameyaw, Meghoma, Dibling.
Booked: Fraser.
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