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Ashton equalled the Six Nations Championship record of six tries held by Shane Williams and Will Greenwood with two further scores in the second half with replacement Danny Care and flanker James Haskell joining in on the act.
Both teams had been forced into late changes with England bringing in Alex Corbisiero for Andrew Sheridan but the young debutant passed his scrummaging test with flying colours against the formidable Martin Castrogiovanni.
Italy boss Nick Mallett's bad luck with half backs again struck with scrum-half Pablo Canavosio, himself an injury replacement, had to be replaced by scrum-half Fabio Semenzato who had little impact on the match.
England supporters did not have to wait long to see the opening try which was almost a mirror image of Ashton's first score against Wales.
Flood, fed by half-back partner Ben Youngs, shot through a gap in the Italian defence and drew Luke McLean's tackle before finding Ashton who ignored Martin Johnson's pre-match instructions by repeating his swan-dive over the line.
Fortunately for the rugby league convert, the ball was grounded safely and Flood converted before exchanging penalties with Mirco Bergamasco.
The Italian winger's second penalty reduced the hosts' lead to four points but that was as good as it got for the Azzurri at Twickenham.
On 24 minutes an England rolling maul made ground down the left, Youngs moved the ball to Shontayne Hape who broke the feeble tackle of Luciano Orquera before offloading to Ashton on his shoulder to touch down. Flood converted for a 17-6 lead.
While Ashton has been in phenomenal scoring form since bursting onto the international scene against France in last year's Six Nations, England's other wing Cueto had not crossed in his previous 18 Tests.
But that drought was broken five minutes later with Flood again the provider for Cueto to dive over with glee.
England were rampant at this point and they got their fourth try through Tindall, whose last international try came four years ago, after a wonderful offload from No.8 Nick Easter.
Flood's fourth conversion of the half gave England a 31-6 advantage.Italy's afternoon went from bad to worse just three minutes into the second half after Castrogiovanni was yellow carded for deliberately preventing Youngs from taking a quick tap penalty.
But just as the prop's spell in the sinbin was due to end, the Italians conceded yet again as replacement Matt Banahan made inroads before Ashton showed his poacher's instinct by snaffling the loose ball and burrowing over.
Flood's conversion was his last act before he was replaced by Jonny Wilkinson as England made a raft of replacements.
One of those fresh faces, scrum-half Care spotted to a gap in the tiring Italian defence to dive over for England's sixth try.
Although replacement hooker Fabio Ongaro bagged a consolation try for the visitors at the back of a rolling maul there was still time for flanker Haskell to barge over before Ashton rounded off his afternoon with another breakaway try.
Replacement fly-half Wilkinson added three conversions but the afternoon belonged to cross-codes convert Ashton who became the first person to score four tries in a Six Nations match.
|England Score Card|
|Mike Tindall (capt)||1||5|
|Italy Score Card|