The Three Lions are ramping up their preparations for this weekend's inaugural tie after flattering to deceive in their pair of friendly matches against Iceland and Bosnia-Herzegovina earlier in the month.
Southgate's side got off to a tepid start against Bosnia before a second-half flurry saw England post a 3-0 victory, but their lack of attacking incisiveness would prove fatal in a shock 1-0 loss to Euro 2016 conquerors Iceland a few days later.
Both John Stones and Marc Guehi emerged as injury concerns after the contest with ankle and head blows respectively, although Southgate quickly played down the seriousness of their issues.
As expected, the duo were both able to take to the field in Tuesday's training session with the remainder of their England teammates, and they are expected to form Southgate's first-choice pairing in Germany due to Harry Maguire's absence.
John Stones absent from England training through illness
However, Stones was unable to take part in Wednesday's practice session having been laid low by an illness, plunging his participation for Sunday's clash with Serbia into some doubt.
The Manchester City man - whose 2023-24 domestic season was also hampered by physical problems - still has ample time to recover from his affliction, and he has not been ruled out of their Group C opener just yet.
Southgate could provide an update on Stones's condition when he faces the media later in the week, but the Three Lions boss is blessed with several alternative options should the ex-Everton defender be forced to sit Sunday's game out.
Lewis Dunk is now training normally again after missing the loss to Iceland due to a knock, while Aston Villa's Ezri Konsa and Liverpool's Joe Gomez can also fill in at the heart of defence.
How is the rest of England's squad shaping up?
While Stones has been forced to take it easy due to his infection, the remainder of Southgate's troops all appear to be unaffected, as there is no indication that the virus has spread through the camp.
As a result, the England boss should be able to put out close to a first-choice XI for the showdown with Dragan Stojkovic's side, including Jude Bellingham and Bukayo Saka, the former of whom sat out both friendly matches.
Meanwhile, Saka was only a late substitute against Iceland but is expected to start on England's right-hand side, although Luke Shaw - who has been nursing a muscle injury since February - is losing his battle to be fit for the opener.
As a result, Kieran Trippier should continue to hold the fort at left-back for the time being, while number eight Trent Alexander-Arnold is expected to start alongside Declan Rice in midfield.
Following Sunday's encounter in Gelsenkirchen, England pit their wits against Denmark at the Deutsche Bank Park on June 20, five days before closing Group C against Slovenia in Cologne.