In July 1623, two more ships arrived, carrying 90 new settlers, among them Leideners, including William Bradford's future wife, Alice. Some of the settlers were unprepared for frontier life and returned to England the next year. In September 1623, another ship carrying settlers destined to refound the failed colony at Weymouth arrived and temporarily stayed at Plymouth. In March 1624, a ship bearing a few additional settlers and the first cattle arrived. A 1627 division of cattle lists 156 colonists divided into twelve lots of thirteen colonists each. Another ship also named the Mayflower arrived in August 1629 with 35 additional members of the Leiden congregation. Ships arrived throughout the period between 1629 and 1630 carrying new settlers; though the exact number is unknown, contemporary documents claimed that by January 1630 the colony had almost 300 people. In 1643 the colony had an estimated 600 males fit for military service, implying a total population of about 2,000. By 1690, on the eve of the dissolution of the colony, the estimated total population of Plymouth County, the most populous, was 3,055 people. It is estimated that the entire population of the colony at the point of its dissolution was around 7,000. For comparison it is estimated that between 1630 and 1640, a period known as the Great Migration, over 20,000 settlers had arrived in Massachusetts Bay Colony alone, and by 1678 the English population of all of New England was estimated to be in the range of 60,000. Despite the fact that Plymouth was the first colony in the region, by the time of its annexation it was much smaller than Massachusetts Bay Colony.
The Little John was on its maiden voyage, having been specially built for the use of the Plymouth Colony pilgrims.
It and the Anne arrived at Plymouth a week apart during the summer of 1623 to find a tiny settlement decimated by illness. Its survivors had struggled to endure many hardships, clinging to the beliefs which had led them to America in search of freedom and self-determination.