St Pauli and Hannover were the first Bundesliga sides to go out of the DFB-Pokal this season as they lost 1-0 to Chemnitz and on penalties against Elversberg respectively.
A goal from Chemnitz's Andreas Richter in the fifth minute was enough to knock the Bundesliga newcomers St Pauli out of the competition, while Hannover struggled at Bavarian side Elversberg, crashing out 5-4 on penalties after the match had ended goalless with Mikael Forssell missing from the spot.
The defeat will put pressure on Reds coach Mirko Slomka, whose side has looked far from impressive in pre-season after only avoiding relegation to the second division on on the final day of last season.
The rest of their top-flight rivals secured their passage to the second round without any problems. Bayer Leverkusen secured the most emphatic win of the day with an 11-1 triumph at Pirmasens, despite only leading 1-0 at half time.
Germany internationals Patrick Helmes and Stefan Kiessling were in scoring mood with three and two goals respectively while Eren Derdiyok left coach Jupp Heynckes a welcome selection headache with a brace after coming on for Helmes.
Borussia Dortmund warmed up for their Europa League play-off on Thursday with a 3-0 win at Wacker Burghausen with Lucas Barrios, Nevan Subotic and Kevin Grosskreutz on target while a goal from Papiss Demba Cisse was enough to see Freiburg through against Oberneuland.
Stuttgart defeated Babelsberg 2-1 while Werder Bremen thrashed Rot Weiss Ahlen 4-0 and Hoffenheim beat Hansa Rostock by the same scoreline and Borussia Monchengladbach defeated Erzgebirge Aue 3-1.
Elsewhere, second division side 1860 Munich came from behind to beat SC Verl 2-1 while Bavarian rivals Augsburg, who were beaten semi-finalists last season, only just scraped past SV Sandhausen. A penalty in stoppage time from Nando Rafael took the game to extra time and Augsburg won 4-1 on penalties.
Extra time was also required for Greuther Furth to beat Eintracht Braunschweig 2-1.
There are 11 more fixtures on Sunday before the round concludes on Monday evening with Bayern Munich's trip to Germania Windeck, the lowest-ranked side in the competition, and Schalke's journey to Aalen.