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Diffusible iodine-based contrast-enhanced computed tomography (diceCT) is an increasingly used digital complement, supplement, or alternative to traditional dissection-based anatomical research. The diceCT protocol, which has evolved and expanded over the past decade, employs passive diffusion of Lugol's iodine (KI3) to increase soft tissue radiodensity and improve structure contrast in the CT or microCT imaging of specimens. The development and application of diceCT has focused largely on specimens under 1 kg, and the varying reporting of methods on studies of both small and large specimens has initiated, but not yet established, an effective diceCT protocol for larger specimens based on monitored experiments of several fundamental variables (e.g., Lugol's iodine concentration, duration, and impacts of Lugol's iodine on tissues). In this study, we have experimentally assessed the efficacy of diceCT protocols for imaging whole-body specimens of the 1–4.5 kg Australian brushtail possum (Trichosurus vulpecula) using sequential CT imaging assessment across experimental conditions. We assessed the impact of varying Lugol's iodine concentration, the presence/absence of skin, solution volume and agitation on tissue radiodensity changes through weekly CT-based monitoring of tissue radiodensities over an 8-week experimental period. We have also quantified tissue volumetric changes across our experiment to assess the impact of diceCT applications on subsequent analyses of imaging datasets. Our results indicate that substantial changes in both soft-tissue radiodensity and soft-tissue volume occur within the first 28 days of Lugol's iodine treatment, followed by a slower rate of progressive soft-tissue radiodensity and volume changes across the experiment duration. Our results demonstrate the negligible benefit of skinning larger specimens to improve solution diffusion, and document significant soft-tissue volumetric changes with high concentration solutions (e.g., 10%) and long-duration exposure (e.g., beyond 5 weeks) that should guide individual diceCT protocol design and/or quantification and analysis for mammal specimens above 1 kg. [ABSTRACT FROM AUTHOR]

Establishing relationships between soft tissues and bones in living species can inform our understanding of functional adaptations in their extinct kin in the absence of direct data on habitual behaviours. The koala and the wombats are the only surviving species of the vombatiform suborder, and represent the extant phylogenetic bracket for dozens of fossil marsupial species across nine families. Here we present the first quantitative architectural data on the forelimb myology of the koala and common wombat, alongside redescriptions based on physical and digital dissections. Despite obvious contrasts in how their forelimbs are used in arboreal (koala) and fossorial (wombat) functional contexts, overall they showed only minor differences in qualitative and quantitative measures of myology. When the architectural properties of antagonistic pairs of muscle groups crossing each joint were compared, the greatest contrasts were seen in muscles crossing the elbow, where the wombat exhibited greater relative emphasis on forceful elbow extension than the koala, indicative of adaptations for digging. Our findings reinforce the importance of the elbow as a useful system to explore when seeking discrimination between habitual patterns of forelimb use in extinct vombatiform species. [ABSTRACT FROM AUTHOR]