The technically gifted attacking midfielder tore a cruciate ligament while playing for Qatari club Al-Duhail in a 5-2 win over Al-Sailiya.
Al-Duhail said in a statement that Msakni ”will be absent for three to four months.”
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Unbeaten during its qualifying campaign, Tunisia is making a return to the World Cup for the first time in 12 years. The Eagles of Carthage will take part in their fifth World Cup but have never got beyond the group stage.