Demystifying Cassiopea species identity in the Florida Keys: Cassiopea xamachana and Cassiopea andromeda coexist in shallow waters.

The phylogeny of the Upside-Down Jellyfish (Cassiopea spp.) has been revised multiple times in its history. This is especially true in the Florida Keys, where much of the Cassiopea stock for research and aquarium trade in the United States are collected. In August 2021, we collected 55 Cassiopea medusae at eight shallow water sites throughout the Florida Keys and sequenced COI, 16S, and 28S genes. Mitochondrial genes demonstrate that the shallow waters in Florida are inhabited by both Cassiopea xamachana and a non-native Cassiopea andromeda lineage, identified in multispecies assemblages at least thrice. While C. xamachana were present at all sites, the C. andromeda-mitotype individuals were present at only a minority of sites. While we cannot confirm hybridization or lack thereof between the C. xamanchana and C. andromeda lineages, these previously unknown multispecies assemblages are a likely root cause for the confusing and disputed COI-based species identities of Cassiopea in the Florida Keys. This also serves as a cautionary note to all Cassiopea researchers to barcode their individuals regardless of the location in which they were collected.