Understanding decay in marine calcifiers: Micro-CT analysis of skeletal structures provides insight into the impacts of a changing climate in marine ecosystems

Calcifying organisms and their exoskeletons support some of the most diverse and economically important ecosystems in our oceans. Under a changing climate, we are beginning to see alterations to the structure and properties of these exoskeletons due to ocean acidification, warming and accelerated rates of bioerosion. Our understanding has grown as a result of using micro-computed tomography (µCT) but its applications in marine biology have not taken full advantage of the technological development in this methodology. We present a significant advancement in the use of this method to studying decalcification in a marine calcifier. We present a detailed workflow on best practice for µCT image processing and analysis of marine calcifiers, designed using coral skeletons subjected to acute, short-term microbial bioerosion. This includes estimating subresolution microporosity and describing pore space morphological characteristics of macroporosity, in perforate and imperforate exoskeletons. These metrics are compared between control and bieroded samples, and are correlated with skeletal hardness as measured by nanoindentation. Our results suggest that using subresolution microporosity analysis improves the spatiotemporal resolution of µCT data and can detect changes not seen in macroporosity, in both perforate and imperforate skeletons. In imperforate samples, the mean size and relative number of pores in the macroporous portion of the images changed significantly where total macroporosity did not. The increased number of pores and higher microporosity are both directly related to a physical weakening of the calcareous exoskeletons of imperforate corals only. In perforate corals, increased macroporosity was accompanied by an overall widening of pore spaces though this did not correlate with sample hardness. These novel techniques complement traditional approaches and in combination demonstrate the potential for using µCT scanning to sensitively track the process of decalcification from a structural and morphological perspective. Importantly, these approaches do not necessarily rely on ultra-high resolution (i.e. single micron) scans and so maintain the accessibility of this technology. The continued optimization of these tools for a variety of marine calcifiers will advance our understanding of the effect of climate change on marine biogenic calcified structures.