Dolphin-derived NETosis results in rapid Toxoplasma gondii tachyzoite ensnarement and different phenotypes of NETs

Toxoplasma gondii is a cosmopolitan zoonotic parasite and nowadays considered as an emerging neozoan pathogen in the marine environment. Cetacean innate immune reactions against T. gondii stages have not yet been investigated. Thus, T. gondii tachyzoites were utilized to trigger neutrophil extracellular traps (NETs) in bottlenose dolphin (Tursiops truncatus) polymorphonuclear neutrophils (PMN). Scanning electron microscopy unveiled T. gondii tachyzoites as potent and rapid inducers of cetacean-derived NETosis. Co-localization of extracellular chromatin with global histones, granulocytic myeloperoxidase and neutrophil elastase confirmed classical characteristics of NETosis. Interestingly, different phenotypes of NETs were induced by tachyzoites resulting in spread, diffuse and aggregated NET formation and moreover, 'anchored' and 'cell free' NETosis was also detected. Current data indicate that cetacean-derived NETosis might represent an early, ancient and well-conserved host innate defense mechanism that not only acts against T. gondii but might also occur in response to other closely related emerging apicomplexan parasites affecting marine cetaceans.