James Magnussen was left looking worryingly human as Australia's heavily-favoured men's 4x100m freestyle relay team flopped, putting huge question marks over his quest for individual glory at the London Olympics.
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In the biggest shock of the meet so far, Australia finished fourth as France overpowered the US to claim the gold medal, with Russia taking third spot.
Magnussen, Matt Targett, Eamon Sullivan and James Roberts were at a loss to explain the performance but denied the world champions had caved to the pressure of favouritism.
On a night when two world records had already fallen, and Australia had won minor medals through Christian Sprenger and Alicia Coutts, "Missile" Magnussen was expected to seriously threaten Cesar Cielo's 100m mark and fire his team to glory with a blistering opening leg.
But he was well off his brilliant best as he clocked 48.03 seconds, almost a second slower than his PB, to be second at the first changeover behind American Nathan Adrian (47.89).
From there Australia failed to recover, despite an encouraging 47.68 swim by veteran Sullivan, and were left shattered on the pool deck as Frenchman Yannick Agnel reeled in US star Ryan Lochte with a scorching 46.74 final leg.
Magnussen appeared devastated when interviewed by Nine commentator Gian Rooney after the race.
"I've got no response really, I don't know what happened," he said, shaking his head, before disappearing behind his teammates as they finished the interview.
World champion Magnussen had been considered a near-certainty to become Australia's first men's Olympic 100m freestyle champion in 44 years, but his rivals will now believe they can beat him when the individual event starts on Tuesday.
"Obviously that is my first big disappointment at international level so I have to regroup," Magnussen later told reporters.
"I knew at the turn I was stinging and it was going to be a long second 50 and that is not usually how it feels for me.
"Obviously it's really disappointing, but it happens.
"I thought I'd be swimming quick tonight."
Magnussen denied the immense pressure on him was a factor but said a slick 47.35 swim in Sunday's heats may have taken "a bit out of the tank", despite feeling easy.
Australian head coach Leigh Nugent said Magnussen and fellow young gun Roberts would have to "get over it and get on with it" for the individual event while Sullivan backed them to do so.
"More than anything, one race doesn't decide your fate for the rest of the meet," Sullivan said.
The disappointment overshadowed some encouraging swims for Australia on the second night of finals.
Sprenger (58.93) produced the swim of his career to take silver in the 100m breaststroke final as South African Cameron van der Burgh clocked 58.46 to take down Brenton Rickard's 2009 world record.
Coutts (56.94) fought home strongly to claim bronze in the women's 100m butterfly final.
But she was no match for American Dana Vollmer, who became the first woman ever to complete the race under 56 seconds as she posted 55.98 to beat Swede Sarah Sjostrom's world record.
Emily Seebohm continued her brilliant form in the 100m backstroke semi-finals, clocking 58.39 to qualify fastest for the final after she set a new Olympic record with 58.23 in the heats.
Belinda Hocking also reached the final while Leisel Jones moved through to the 100m breaststroke decider and Hayden Stoeckel progressed in the 100m backstroke semis.
Thomas Fraser-Holmes booked a dream berth in the men's 200m final where he will face Lochte, Agnel, Chinese star Sun Yang and German world record holder Paul Biedermann.