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Busquets and his La Roja teammates drew 0-0 with Portugal in a friendly over the weekend but it emerged that the Barcelona man had been infected by the virus when he returned a positive test on Sunday.
The rest of the squad and coaches all tested negative but because they were in close contact with Busquets, they too are required to self-isolate and have therefore been training alone.
It means that the scheduled final Euro 2020 warm-up fixture with Lithuania in Leganes will see Spain's Under 21 players feature instead - while manager Luis De la Fuente will take charge instead of senior boss Luis Enrique.
In addition, Spain have created a "parallel bubble" in the event that further positive tests arrive and they need to call upon replacements.
:rotating_light: OFICIAL ' El guardameta @kepa_46 se une a los futbolistas convocados para ejercitarse en la burbuja paralela de la @SeFutbol en Las Rozas.
:arrow_right: Los entrenamientos de los jugadores invitados comenzarán hoy a las 11:00 horas.
:information_source: https://t.co/zJ8nunXpGM#SomosEspaña #EURO2020 pic.twitter.com/OClAWMDKde
- Selección Española de Fútbol (@SeFutbol) June 8, 2021
On Monday morning, it was announced that Chelsea goalkeeper Kepa Arrizabalaga has become the sixth member of that bubble, joining Leeds forward Rodrigo Moreno, West Ham's Pablo Fornals, Valencia midfielder Carlos Soler, Celta Vigo winger Brais Mendez and Villarreal defender Raul Albiol in their Las Rozas training camp.
Spain have called up Carlos Soler, Brais Méndez, Pablo Fornals and Rodrigo Moreno to train in a separate bubble after Sergio Busquets tested positive for coronavirus #esp #EURO2020 pic.twitter.com/zgIwbq0ixg
- Goal (@goal) June 7, 2021
The RFEF said: "In addition, once the game against Lithuania is over, a small group of footballers yet to be confirmed will join the training sessions of the senior team under the same conditions as the previous ones.
"They will train at different times and will not mix in any case with those already called up."
Spain kick off their Euros campaign against Sweden in Group E on 14 June and will be without skipper Busquets due to his mandatory quarantine period.
The 32-year-old could be back for the game against Poland on 19 June and Spain will be praying for no more positive cases amongst their squad.
Meanwhile, the country's Sports Minister Jose Manuel Manuel Rodriguez has said the squad will be given a vaccine shot tomorow.
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