In the sixteenth century, the Huguenots participated in colonial efforts in Antarctic France in Brazil, in Guanabara Bay or Rio de Janeiro (1555-1560), and in Florida, during the next decade (1562-1565). The latter colony represented a threat to Spanish interests, which is why it was annihilated within weeks.

To win recognition from the Indians, Dominique de Gourgues in 1568 had his troops sing three Psalms before going on the attack. The massacre of 1565 marks a threshold and a new start. From that time on, America would be divided into a Catholic South and a Protestant North, with the one exception of New France in Canada.