Vicenza, champions in 1997, now reside in the third tier of Italian football and were defeated 2-1 at home by Bassano Virtus after taking an early lead through Piergiuseppe Maritato.
That was before goals in the last 15 minutes from Luca Cattaneo and Filippo Stevanin turned the tie in the visitors' favour as the home side fell at the first hurdle.
Meanwhile, Venezia, who lifted the trophy way back in 1941, had a much more straightforward day as they breezed past Taranto 5-1, with Tommaso Bellazzini scoring twice, either side of half-time.
Other comfortable victors on Sunday were Savona, who thumped Terracina Calcio 6-0 in a game that including a brace from Angelo De Martis, and Como, 5-0 winners over Matelica Calcio.
Lecce were similarly emphatic, trouncing Foligno 5-0 at home while Pisa saw off RapalloBogliasco 4-0.
Much more closely fought was the clash between L'Aquila and Alto Vicentino, which saw the former advance 2-1 in extra-time thanks to Lorenzo Del Pinto's 105th-minute winner.
Other tight affairs saw Casertana edge former Serie A outfit Reggina 1-0, Alessandria squeeze past Salernitana by the same scoreline and Monza overcome Olginatese 2-1.
Nuova Cosenza came back from two goals down away at Cremonese to level at 2-2, only for Radoslav Kirilov to win it for the hosts midway through the second half.
A late brace from Manuel Fischnaller helped Sudtirol defeat Teramo 3-2, while Benevento and Catanzaro were both 2-0 winners, over Correggese and Akragas respectively.
Renate were 1-0 victors over AlbinoLeffe, as were FeralpiSalo over Santarcangelo, while elsewhere, Pontedera saw off Messina 3-1 and Juve Stabia got the better of Prato 1-0.
