Cubozoan jellyfish pose a risk of envenomation to humans and a threat to many businesses, yet crucial gaps exist in determining threats to stakeholders and understanding their ecology. Environmental DNA (eDNA) provides a cost-effective method for detection that is less labour intensive and provides a higher probability of detection. The objective of this study was to develop, optimise and trial the use of eDNA to detect the Australian box jellyfish, Chironex fleckeri. This species was the focus of this study as it is known to have the strongest venom of any cubozoan; it is responsible for more than 200 recorded deaths in the Indo-Pacific region. Further, its ecology is poorly known. Herein, a specific and sensitive probe-based assay, multiplexed with an endogenous control assay, was developed, and successfully utilised to detect the deadly jellyfish species and differentiate them from closely related taxa. A rapid eDNA decay rate of greater than 99% within 27 h was found with no detectable influence from temperature. The robustness of the technique indicates that it will be of high utility for detection and to address knowledge gaps in the ecology of C. fleckeri; further, it has broad applicability to other types of zooplankton.

Understanding the hierarchy of populations from the scale of metapopulations to mesopopulations and member local populations is fundamental to understanding the population dynamics of any species. Jellyfish by definition are planktonic and it would be assumed that connectivity would be high among local populations, and that populations would minimally vary in both ecological and genetic clade-level differences over broad spatial scales (i.e., hundreds to thousands of km). Although data exists on the connectivity of scyphozoan jellyfish, there are few data on cubozoans. Cubozoans are capable swimmers and have more complex and sophisticated visual abilities than scyphozoans. We predict, therefore, that cubozoans have the potential to have finer spatial scale differences in population structure than their relatives, the scyphozoans. Here we review the data available on the population structures of scyphozoans and what is known about cubozoans. The evidence from realized connectivity and estimates of potential connectivity for scyphozoans indicates the following. Some jellyfish taxa have a large metapopulation and very large stocks (>1000 s of km), while others have clade-level differences on the scale of tens of km. Data on distributions, genetics of medusa and polyps, statolith shape, elemental chemistry of statoliths and biophysical modelling of connectivity suggest that some of the ~50 species of cubozoans have populations of surprisingly small spatial scales and low levels of connectivity. Despite their classification as plankton, therefore, some scyphozoans and cubozoans have stocks of small spatial scales. Causal factors that influence the population structure in many taxa include the distribution of polyps, behavior of medusa, local geomorphology and hydrodynamics. Finally, the resolution of patterns of connectivity and population structures will be greatest when multiple methods are used.

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