Striker David Suazo will not recover in time to face Chile on Wednesday according to a Honduras trainer.
The 30-year-old Inter Milan forward is facing a race against the clock to shake off a muscular problem before the Group H clash, in Honduras' second-ever World Cup campaign.
Suazo had been touch-and-go for the game, but Carlos Velasco says from the "physical point of view, he isn't in condition" to play.
Manager Reinaldo Rueda will have the final say on whether Suazo will feature against the South Americans, and there is also doubt on the fitness of Wilson Palacios.
Los Catrachos have a largely inexperienced squad in South Africa, but Rueda hopes the team spirit and desire will carry them through the tournament.
He said: "Honduras must compensate for the lack of global experience with a desire and willingness to do things right, with motivation, with the camaraderie between them."
Honduras also face games against Switzerland and European champions Spain as they look to reach the knockout stage.