The Uruguayan hosts are unblemished on their turf in this season's competition, and they seek to continue that run against a club that struggled on the road in the group phase.
Match preview
Penarol ended second in Group G, but the Uruguayan outfit could have exited had they not won every group-stage match in front of their supporters.
Decano beat Caracas (5-0), Atletico Mineiro (2-0) and Rosario Central (2-1) on home turf, with their defensive strength seeing them let in one goal in three matches, a statistic that bodes well if continued in the knockout rounds.
On the contrary, Manyas' away record was underwhelming, with two defeats and a victory nearly costing the Uruguayan side their place in the knockout stages.
Diego Aguirre's men seek to continue their home run on Wednesday, keen to put themselves in a strong position heading into the reverse meeting in La Paz.
With three goals scored and none conceded on their turf in their recent two home league games, Aguirre will back his side to keep that run going against the Bolivian visitors.
While Penarol's road results were below par, Tigre's were almost non-existent as they secured a solitary point from an available nine on their travels, notching a point at Huachipato.
The Strongest relied on their equally unblemished record on home soil where they netted seven times and conceded none, and they go into the round of 16 first leg keen to return with a positive result that keeps the tie in the balance.
While the Group C winners head into the tie's first fixture undefeated in four since a 5-2 league thrashing at the hands of GV San Jose, it remains to be seen if their poor road results from the group stage continue into the knockouts.
Ismael Rescalvo's team have also had fewer days to recuperate and prepare for Wednesday's match compared to their hosts whose last fixture came on August 4 while the Bolivian outfit were last in league action four days later.
However, with the last-16 first-leg match six days after their 2-1 victory at Real Santa Cruz, the Strongest should not be excessively worried about accumulated fatigue and little recovery time.
Penarol Copa Libertadores form:
L
W
L
W
W
W
Penarol form (all competitions):
D
D
W
W
W
D
The Strongest Copa Libertadores form:
W
L
D
W
W
L
Team News
Maximiliano Silvera is the home side's undoubted leading match-winner, having scored five group-stage goals in six appearances, three more than Leonardo Fernandez's two.
However, Fernandez's creativity saw him set up three goals for teammates while fashioning four big chances, with Javier Cabrera — two assists, three big chances created — following closely behind.
Ignacio Sosa featured in four of Panarol's six home fixtures but is seemingly out of Wednesday's match due to a muscle problem.
The Strongest have no reported injury-enforced absentees in midweek, giving Rescalvo a healthy squad for their visit to Uruguay.
Unlike their hosts, Tigre spread the goals around in the group stage, with Enrique Triverio and Luciano Ursino notching two each for the Bolivians.
Penarol possible starting lineup:
Aguerre; Milans, Coelho, Rodriguez, Olivera; Cabrera, Garcia, Mendez, Hernandez; Silvera, Fernandez
The Strongest possible starting lineup:
Gutierrez; Bustos, Aimar, Jusino, Chura; Cuellar, Ursino; Arrascaita, Ortega, Ayarza; Triverio