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Horseshoe crabs are a group of marine chelicerates that contain only four extant species, some of which are endangered. Their hemolymph has been widely used in medical applications for endotoxin detection. Nevertheless, there is limited information on the profiles of their hemolymph proteins and their gut microbial diversity. In this study, we performed the first detailed investigation of the hemolymph proteomics and gut microbiota of two Asian horseshoe crabs  Tachypleus tridentatus  and  Carcinoscorpius rotundicauda . Among the identified proteins being cataloged in the juvenile and adult hemolymph, unexpectedly, sesquiterpenoid signaling pathway proteins including Heat shock protein 83 (HSP83), Chd64, and a juvenile hormone binding protein (JHBP) were revealed. This provides evidence for the presence of functional sesquiterpenoid hormonal systems in these marine chelicerates. consumption of certain horseshoe crab species often leads to tetrodotoxin poisoning and the horseshoe crab is thought to possess a tetrodotoxin resistance mechanism. As such, sodium channels were analyzed and found to have critical amino acid residues that are similar to the toxin resistant pufferfish sodium channels. The source of the toxin is unknown so we investigated the gut microbiota, and found that  Clostridium  and  Vibrio  were the most dominant bacteria in  T. tridentatus  and  C. rotundicauda , respectively. Together, this study provides a framework for further understanding of sesquiterpenoids and gut microbiota of these marine chelicerates.

Hemolymph Proteomics and Gut Microbiota of Horseshoe Crabs Tachypleus tridentatus and Carcinoscorpius rotundicauda