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Traditional European heavyweights Chelsea and Liverpool headline a quartet of English clubs harbouring Europa League dreams as the round of 32 gets underway on Thursday.
Two-time champions Tottenham also join Newcastle among the Premier League hopefuls seeking to make their mark in the competition's knockout phase.
Defending European champions Chelsea infamously made history when they became the first Champions League title-holders to exit the competition in the group stage, but the Blues are now aiming to enter the record books in a far more positive light.
"It is true the Europa League is not the same as the Champions League because that is the top but the Europa League is a big competition, an important competition, and now we have the chance to win it. Never in the history of Chelsea have we won this cup and we are going to try to do it," said Spanish defender Cesar Azpilicueta.
The Londoners travel to Sparta Prague on Thursday to face a Czech team playing their first competitive fixture since the winter break.
Rafael Benitez's stuttering side have won just once in five outings -- a 4-1 pasting of strugglers Wigan on Saturday -- but Sparta goalkeeper Tomas Vaclik conceded the Czech outfit still have a monumental challenge on their hands.
"Even if Chelsea are not having their best period right now, they have the winning mentality of a big club and self-confidence. To succeed, we'll have to play our best football."
Liverpool's quest for a record fourth title takes them to Saint Petersburg for a first-leg tie against double defending Russian Premier League champions Zenit.
Liverpool lie ninth in the table after a dismal 2-0 home defeat to West Brom on Monday, but first-year boss Brendan Rodgers hopes his players can quickly put that setback behind them against the 2008 Europa League winners.
"We just have to keep striving and look towards the next game. We've got the Europa League game on Thursday night, which is a good opportunity for us to progress in a competition."
Following their shock FA Cup exit to third-tier Oldham, the tournament represents the Reds' last realistic chance of silverware this season.
Meanwhile, Tottenham host Ligue 1 title challengers Lyon at White Hart Lane with Spurs goalkeeper Hugo Lloris set to face his former teammates for the first time since his August departure.
"It's not easy for me because I have a good relationship with all my ex-teammates but that is football, that is the life. On the pitch there are no friends. We play for our team and give all the best for our team," said Lloris.
Spurs, the inaugural winners of the competition in 1972 as well as 1984, are undefeated in 10 Premier League outings, in stark contrast to Lyon who are coming off successive league defeats for the first time since 2011, denting their pursuit of a first Ligue 1 crown since 2008.
The north Londoners will, however, be without leading striker Jermain Defoe with the England international sidelined through injury.
Buoyed by their latest French recruits, Newcastle go into their clash with Ukrainian side Metalist Kharkiv at St. James' Park -- where they are unbeaten in 11 European home games -- in confident mood.
"The new injection of players has given everybody a lift at the club and you can see that on the pitch when we go out and play," said Magpies defender James Perch, referring to the January arrivals of Mapou Yanga-Mbiwa, Moussa Sissoko, Mathieu Debuchy, Yoan Gouffran and Massadio Haidara.
Other notable ties include holders Atletico Madrid hosting Russian outfit Rubin Kazan, while three-time winners Inter Milan are at home to Romania's CFR Cluj and Napoli entertain Czech table-toppers Viktoria Plzen.