Using hydrofluoric acid for morphological investigations of Zoanthids (Cnidaria: Anthozoa): a critical assessment of methodology and necessity.

Zoanthids comprise an order of benthic, generally colonial cnidarians, which can usually be distinguished from other hexacorallians by embedded sand and detritus in their mesoglea to help strengthen their structure. These animals are becoming increasingly important research subjects in biochemistry and other research fields. Their inclusion of both calcium and silica results in the need for both decalcification and desilification for internal morphological examinations. Since the methodology of hydrofluoric acid (HF) desilification has rarely been documented in zoanthids, histological surveys for zoanthid taxonomy have often been abandoned and their taxonomy is often problematic. Recent investigations utilizing molecular methods have brought a clearer understanding of zoanthid diversity, but standardization of HF treatments are still needed to provide a link between molecular and more traditional techniques, and to properly examine specimens for which molecular methods may not be an option (e.g., formalin-preserved specimens, etc.). Here, we use both "straight" HF and, for the first time with zoanthids, buffered HF (BHF) treatments at different treatment lengths (1-48 h) on polyps from three different species of zoanthids for histological examination. Section conditions were judged based on the presence/absence of embedded detritus, drag marks, and tissue condition. Results show that the BHF treatment resulted in slightly better tissue conditions for all specimens, and suggest that desilification works well regardless of treatment time for species with smaller (polyp diameter <0.5 cm), less heavily encrusted polyps. Desilification of heavily encrusted Palythoa mutuki polyps were still problematic, with at least 24 h treatment needed. To aid future research, we provide guidelines for HF treatments of zoanthid specimens.