In 1524, Italian explorer Giovanni da Verrazzano, (1485–1528), in service of the French crown, (famous for sailing through and thereafter naming the entrance to New York Bay as the "Verrazzano Narrows", including now in the 20th century, a suspension bridge also named for him) sailed past the Chesapeake, but did not enter the Bay. Spanish explorer Lucas Vásquez de Ayllón sent an expedition out from Hispaniola in 1525 that reached the mouths of the Chesapeake and Delaware Bays. It may have been the first European expedition to explore parts of the Chesapeake Bay, which the Spaniards called "Bahía de Santa María" ("Bay of St. Mary") or "Bahía de Madre de Dios."("Bay of the Mother of God") De Ayllón established a short-lived Spanish mission settlement, San Miguel de Gualdape, in 1526 along the Atlantic coast. Many scholars doubt the assertion that it was as far north as the Chesapeake; most place it in present-day Georgia's Sapelo Island. In 1573, Pedro Menéndez de Márquez, the governor of Spanish Florida, conducted further exploration of the Chesapeake. In 1570, Spanish Jesuits established the short-lived Ajacan Mission on one of the Chesapeake tributaries in present-day Virginia.
The arrival of English colonists under Sir Walter Raleigh and Humphrey Gilbert in the late 16th century to found a colony, later settled at Roanoke Island (off the present-day coast of North Carolina) for the Virginia Company, marked the first time that the English approached the gates to the Chesapeake Bay between the capes of Cape Charles and Cape Henry. Three decades later, in 1607, Europeans again entered the Bay. Captain John Smith of England explored and mapped the Bay between 1607 and 1609, resulting in the publication in 1612 back in the British Isles of "A Map of Virginia". Smith wrote in his journal: "Heaven and earth have never agreed better to frame a place for mans habitation." The Captain John Smith Chesapeake National Historic Trail, the first designated "all-water" National Historic Trail in the US, was established in 2006 by the National Park Service. The trail follows the route of Smiths historic 17th-century voyage. Because of economic hardships and civil strife in the "Mother Land", there was a mass migration of southern English Cavaliers and their servants to the Chesapeake Bay region between 1640 and 1675, to both of the new colonies of the Province of Virginia and the Province of Maryland.