Ballasting and deballasting of shipping vessels in foreign ports have been reported worldwide as a vector of introduction of non-native aquatic plants and animals. Recently, attention has turned to ballast water as a factor in the global increase of harmful algal blooms (HABs). Many species of microalgae, including harmful dinoflagellate species, can remain viable for months in dormant benthic stages (cysts) in ballast sediments. Over a period of four years, we surveyed ballast water and sediment of ships docked in two ports of Tampa Bay, Florida, USA. Sampling conditions encountered while sampling ballast water and sediments were vastly different between vessels. Since no single sample collection protocol could be applied, existing methods for sampling ballast were modified and new methods created to reduce time and labor necessary for the collection of high-quality, qualitative samples. Five methods were refined or developed, including one that allowed for a directed intake of water and sediments. From 63 samples, 1,633 dinoflagellate cysts and cyst-like cells were recovered. A native, cyst-forming, harmful dinoflagellate, Alexandrium balechii (Steidinger) F. J. R. Taylor, was collected, isolated, and cultured from the same vessel six months apart, indicating that ships exchanging ballast water in Tampa Bay have the potential to transport HAB species to other ports with similar ecologies, exposing them to non-native, potentially toxic blooms.