Spain booked a quarter-final date with Paraguay by beating Portugal 1-0 in Cape Town, a 63rd-minute David Villa goal proving the difference between the two sides.
Paraguay reached the World Cup quarter-finals for the first time as they edged Japan 5-3 in a penalty shoot-out after a drab goalless draw in Pretoria.
Brazil knocked Chile out of the World Cup for the third time as Juan, Luis Fabiano and Robinho scored to set up a quarter-final showdown with the Netherlands.
Germany's youthful side delivered a striking statement of intent by overwhelming old rivals England 4-1 through goals from Miroslav Klose, Lukas Podolski and Thomas Muller (2).
Asamoah Gyan scored three minutes into extra time to secure a historic place in the World Cup quarter-finals for Ghana after a dramatic 2-1 victory over USA in Rustenburg.
Goals from David Villa and Andres Iniesta earned Spain a 2-1 win over ten-man Chile and a last-16 meeting with Portugal, while the losers also made it through despite defeat.
Ivory Coast ran out comfortable 3-0 winners over Korea DPR in Nelspruit, but Portugal's draw against Group G winners Brazil confirmed the African team's elimination.
Portugal joined Brazil in the Round of 16 as Group G runners-up after the teams played out a hard-fought, if at times ill-tempered goalless draw in Durban.
The Netherlands maintained their 100 per cent record in Group E with a 2-1 win over Cameroon in a match which saw Arjen Robben make a successful comeback from injury.
Paraguay finished top of Group F following a scoreless draw with New Zealand in Polokwane that sent the undefeated All Whites home with their heads held high.
Australia hit second-half goals from Tim Cahill and Brett Holman to leap Serbia in the Group D standings, but were still eliminated on goal difference by Ghana.
Germany edged Ghana 1-0 to win Group D and secure a meeting with England, while their victims also advanced to the Round of 16, where they will face USA.
Argentina won their third straight game at the 2010 World Cup to top Group B and eliminate a Greek side that paid for their almost complete lack of ambition.
An up-and-down thriller in Durban eventually ended in a 2-2 draw between Korea Republic and Nigeria, sending the Asians into a last-16 meeting with Uruguay.
Uruguay finished top of Group A thanks to a hard-fought 1-0 win over Mexico at the Royal Bafokeng Stadium in Rustenburg, but the defeat was ultimately sufficient for El Tri to join their conquerors in the Round of 16.
South Africa ended their Group A campaign on a high note with a battling win over France, as the hosts were eliminated from the World Cup on goal difference.
David Villa scored twice but missed a penalty as Spain bounced back from defeat by Switzerland with a 2-0 victory over Group H rivals Honduras at Ellis Park.