Bouzas Maneiro will next face either Ana Bogdan of Romania or Spanish compatriot Cristina Bucsa for a spot in the last 32
The 21-year-old Spaniard secured a 6-4, 6-2 victory over the sixth-ranked Czech player, who had previously made history as the first unseeded woman to win the title at the All England Club last year.
The last time a defending champion was defeated in the opening round was in 1994 when German legend Steffi Graf lost to Lori McNeil.
Until Tuesday, Bouzas Maneiro had never defeated a top-40 player, and her only other grass-court match this year was a first-round loss in a minor event in Italy.
“This is one of the most important moments in my life and career," said Bouzas Maneiro. "She is one of the best players in the world, so there was no pressure on me. I could just enjoy the moment and play freely,”.
She added, "I'm really surprised with myself. I was a little bit nervous at the start, but the atmosphere was so nice that I felt more comfortable. I felt at home."
Bouzas Maneiro will next face either Ana Bogdan of Romania or Spanish compatriot Cristina Bucsa for a spot in the last 32.
Vondrousova had arrived in London as a fitness concern after injuring her right hip during a fall at the Berlin grass-court tournament last month.
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