Months after a storm separated the sloop Three Brothers from the first fleet of English colonists who would found Charles Towne in April 1670, the lost vessel put in near the Spanish mission of Santa Catalina de Guale (St. Catherine’s Island, Georgia) in hopes of procuring much-needed supplies. The initial encounter between the strangers proved cordial, prompting a dozen or so English colonists to go ashore to trade and do laundry. They would not return. The Spanish later demanded the surrender of those remaining onboard the sloop and opened fire on it when they refused. The hostilities at Santa Catalina represented the first diplomatic incident between the fledgling Carolina colony and Spanish Florida. This presentation places the misadventures of the Three Brothers into historical context, first exploring the origins of the greater Anglo-Spanish rivalry, before addressing how it manifested itself in the New World after the English arrived to challenge Spanish dominion.