Defender Elderson Echiejile will miss the World Cup because of an injury suffered in a warm-up match last week, Nigerian officials said on Sunday.
The left back from French club Monaco tore a hamstring in the friendly draw with Greece in Philadelphia on Tuesday and will need a minimum of three weeks to recover.
"We don't have that sort of time to play with," coach Stephen Keshi told reporters after Saturday's 2-1 loss to the United States in Nigeria's last warm-up match before departing for Brazil.
Fellow defender Kenneth Omeruo and winger Ahmed Musa have been passed fit despite both sitting out the defeat by the Americans in Jacksonville.
Nigeria are expected to announce midfielder Ejike Uzoenyi as Echiejile's replacement.
Uzoenyi is from the South African club Mamelodi Sundowns but has been on loan at Enugu Rangers over the last six months.
Nigeria are in Group F with Iran, Bosnia and Herzegovina and Argentina in the World Cup.
(Reporting by Mark Gleeson in Cape Town; Editing by Sudipto Ganguly)