Good relations with the Indians eventually soured and by the following spring the colonists were close to starvation. Twice mutinous parties had sailed off to make their own fortune and some were eventually captured by the Spanish, revealing the presence of the French colony. The remaining colonists, having failed to discover silver or gold, were about to leave Florida in August 1565, when they spotted sails on the horizon. Ribault had arrived with a relief expedition of supplies and 600 soldiers and settlers, including more women and some children.

On learning of Ribault's departure for Florida, Philip II of Spain sent Admiral Pedro Menendez to dislodge the French. Initially rebuffed off the coast, Menendez established a base to the south at St. Augustine. Ribault sailed down the coast seeking to attack the Spanish, but his ships were scattered by a hurricane and beached far to the south. Seizing the opportunity, Menendez marched north with 500 soldiers to attack the weakly guarded colony. Early in the morning of September 20, his troops massacred 140 settlers, sparing only about 60 women and children. Forty to fifty others, including Laudonniere, escaped and sailed for France. Menendez next marched south and found the shipwrecked Frenchmen, Ribault among them. They threw themselves on his mercy, but to Menendez they were heretics and enemies of his king. At a place later named Matanzas (Slaughter), he put to the sword about 350 men-all but those professing to be Catholics and a few musicians.

French revenge was enacted in April 1568 by Dominique de Gourgues. He attacked and burned the fort, killing anyone who didn't escape, then sailed home. Spain rebuilt the fort only to abandon it in 1569, but France never again strongly challenged Spanish claims in North America.

In Florida, both France and Spain hoped to claim their piece of the"new world." By the time the French planted their settlement at La Caroline, Spain was entrenched in South and Central America and its sea routes through the Caribbean were well established, as shown on the map below. Spanish ships bearing gold and silver from the mines of Mexico and Peru stopped at Havana before sailing for Spain. They rode the Gulf Stream through the Bahama Channel (now the Straits of Florida) and up the southeastern coast of North America. The Spanish were uneasy about a French settlement because their treasure ships, while they followed the Florida coast, could be easy prey for suspected French raiders in their nearby haven at La Caroline.

"They be all naked and of goodly stature, mighty, faire and as well shapen. . . as any people in all the worlde, very gentill, curtious and of a good nature. . . the men be of tawny collour, hawke nosed and of a pleasant countenance. . . the women be well favored and modest."

The Timucuans looked to the water for sustenance, settling along rivers or near the coast. (Their prehistoric ancestors are called "People of the Shell Mounds.") Besides collecting shellfish and fishing, they hunted and gathered in the forests and swamps and planted maize, squash, and beans. In their often pallisaded villages, they lived in circular dwellings with conical palm-thatched roofs and walls of woven vines caulked with clay. Ceremonial squares in the larger villages were the scenes of festivals, dances, and religious ceremonies.

Related villages formed a loose political confederation under a head chief. In this caste society commoners paid deference to a hereditary elite, at the pinnacle of which sat the chief. Wealth and title were inherited within clans through the mother's brother.

The Timucuans helped their new neighbors adapt to conditions in the "new world"— sharing food and even helping them build a village and fort. The French, well aware of their minority status, initially made every effort to avoid alienating local tribes. Only when starvation threatened did this policy unravel. Mistrust turned to armed conflict, and the brief period of harmony between French and Indian came to an end. Yet the Timucuans apparently remained neutral during the attack by the Spanish against the French fort in 1565 and actively assisted De Gourgues's forces in the successful French recapture of the fort in 1568.

The Florida tribes could not long survive contact with Europeans. After driving out the French, the Spanish imposed tribute on the Timucuans and forced them into missions. Devastated by European disease and attacks by other Indians, the Timucuan culture rapidly disintegrated. From a population possibly numbering tens of thousands at the time of contact, only an estimated 550 Timucuans were still alive in 1698. Today there are no known Native Americans who call themselves Timucuan.

Timucuans hold an early place in the European consciousness of Native Americans. French colonist Jacques le Moyne's sketches of Timucuan ceremonies and customs provided Europeans with some of their first views of Native Americans. Franciscan priest Francisco Pareja's translation of a set of catechisms and confessionals from Spanish into Timucuan in 1612 was the first translation involving a Native American language.

The original site of La Caroline no longer exists. It was washed away after the river channel was deepened and widened in the 1880's. The construction of the fort exhibit was based on a 15th-century sketch by Jacques le Moyne, the colony's artist and mapmaker.