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Volumetric data provide unprecedented structural insight to the reproductive tract and add vital anatomical context to the relationships between organs. The morphology of the female reproductive tract in non-avian reptiles varies between species, corresponding to a broad range of reproductive modes and providing valuable insight to comparative investigations of reproductive anatomy. However, reproductive studies in reptilian models, such as the brown anole studied here, have historically relied on histological methods to understand the anatomy. While these methods are highly effective for characterizing the cell types present in each organ, histological methods lose the 3D relationships between images and leave the architecture of the organ system poorly understood. We present the first comprehensive volumetric analyses of the female brown anole reproductive tract using two non-invasive, non-destructive imaging modalities: micro-computed tomography (microCT) and optical coherence tomography (OCT). Both are specialized imaging technologies that facilitate high-throughput imaging and preserve three-dimensional information. This study represents the first time that microCT has been used to study all reproductive organs in this species and the very first time that OCT has been applied to this species. We show how the non-destructive volumetric imaging provided by each modality reveals anatomical context including orientation and relationships between reproductive organs of the anole lizard. In addition to broad patterns of morphology, both imaging modalities provide the high resolution necessary to capture details and key anatomical features of each organ. We demonstrate that classic histological features can be appreciated within whole-organ architecture in volumetric imaging using microCT and OCT, providing the complementary information necessary to understand the relationships between tissues and organs in the reproductive system. This side-by-side imaging analysis using microCT and OCT allows us to evaluate the specific advantages and limitations of these two methods for the female reptile reproductive system. Summary Sentence Two volumetric imaging modalities, contrast-enhanced micro-computed tomography and optical coherence tomography, facilitate visualization of the macro- and micro- architecture of the reproductive tract of the female brown anole, Anolis sagrei. Graphical Abstract

Background Post-mortem imaging has become indispensable in marine mammal research, offering non-destructive alternatives to conventional necropsy. While micro-computed tomography (microCT) provides high resolution for small specimens, its utility in cetacean fetal studies is limited by long scanning time, accessibility, and high cost. Diffusible iodine-based contrast-enhanced CT (diceCT) using clinical-grade scanners presents a potential solution, particularly for endangered species like the narrow-ridged finless porpoise ( Neophocaena asiaeorientalis ) (NRFP), where fetal specimens are exceptionally rare. This study pioneers the application of clinical CT-based diceCT for NRFP fetal imaging, addressing the gap in developmental morphology research. Results A 41 cm NRFP fetus was recovered from a stranded pregnant female and preserved for iodine-enhanced CT imaging. The specimen underwent 22 weeks of staining in 1% iodine-ethanol solution, with weekly solution replacement and biweekly clinical CT scans (100 kVp, 300 mAs, 0.3 mm slice thickness) to monitor progression. Complete staining was confirmed by clear radiographic differentiation between adjacent tissue, variance of Hounsfield unit (HU) values of the specimen, and a plateau in the signal-to-noise ratio (SNR) among all scans within the liver. High-resolution 3D reconstructions successfully visualized the gastrointestinal tract, urogenital structures, and cardiovascular network, with preserved spatial relationships. Conclusions Clinical-grade diceCT provides a practical alternative to microCT for cetacean fetal imaging by achieving organ-scale resolution. The protocol enabled comprehensive 3D morphological analysis without tissue destruction. The archived dataset and destained specimen offer enduring value for conservation research, education, and method development.

Abstract Background Cat breeders observed a frequent occurrence of internal hydrocephalus in Persian cats with extreme brachycephalic head morphology. Objective To investigate a possible relationship among the grade of brachycephaly, ventricular dilatation, and skull dysmorphologies in Persian cats. Animals 92 Persian-, 10 Domestic shorthair cats. Methods The grade of brachycephaly was determined on skull models based on CT datasets. Cranial measurements were examined with regard to a possible correlation with relative ventricular volume, and cranial capacity. Persians with high (peke-face Persians) and lower grades of brachycephaly (doll-face Persians) were investigated for the presence of skull dysmorphologies. Results The mean cranial index of the peke-face Persians (0.97 ± 0.14) was significantly higher than the mean cranial index of doll-face Persians (0.66 ± 0.04; P < 0.001). Peke-face Persians had a lower relative nasal bone length (0.15 ± 0.04) compared to doll-face (0.29 ± 0.08; P < 0.001). The endocranial volume was significantly lower in doll-face than peke-face Persians (89.6 ± 1.27% versus 91.76 ± 2.07%; P < 0.001). The cranial index was significantly correlated with this variable (Spearman's r: 0.7; P < 0.0001). Mean ventricle: Brain ratio of the peke-face group (0.159 ± 0.14) was significantly higher compared to doll-face Persians (0.015 ± 0.01; P < 0.001). Conclusion and Clinical Relevance High grades of brachycephaly are also associated with malformations of the calvarial and facial bones as well as dental malformations. As these dysmorphologies can affect animal welfare, the selection for extreme forms of brachycephaly in Persian cats should be reconsidered.

Clinically, the rodent thorax is important because of the variety of problems that may affect the heart, lungs, and other thoracic structures. Syrian hamsters are the most common pet and experimental hamster species. Sectional imaging of small mammals is widely increasing in use for clinical and research settings; however, no reports on the thoracic sectional imaging anatomy in this species have been made. Therefore, this study aimed at evaluating micro-computed tomography (MCT), magnetic resonance image (MRI), and gross sections of the Syrian hamster thorax. Eight healthy adult male and female Syrian hamsters were examined. Anesthetized hamsters were evaluated by MCT and 3 Tesla MRI. After imaging, the frozen slab sections were photographed, and identified anatomic structures were matched with structures on corresponding MCT and/or MRI images. Clinically relevant anatomic features of the Syrian hamster thorax that were identified on transverse and sagittal anatomic sections could be discerned on the corresponding MCT and MRI scans. The three matched images were exhibited the following features: (1) the narrow thoracic cavity and small lung space, (2) the heart positioning ventral and medial to lung, (3) heart ventricles locating towards the left side, and (4) parallel cardiac position relative to the sternum. The obtained results were similar to those have only been reported in rabbits, regardless of heart orientation and lung lobe covering the heart. The results of this study support the use of MCT and MRI as diagnostic tools in Syrian hamsters and provide reference values for the clinically relevant anatomic structures of thorax.

The semicircular canals (SCs) of the inner ear detect angular acceleration and are located in the bony labyrinth of the petrosal bone. Based on high-resolution computed tomography, we created a size-independent database of the bony labyrinth of 50 mammalian species especially rodents of the squirrel-related clade comprising taxa with fossorial, arboreal and gliding adaptations. Our sampling also includes gliding marsupials, actively flying bats, the arboreal tree shrew and subterranean species. The morphometric anatomy of the SCs was correlated to the locomotion mode. Even if the phylogenetic signal cannot entirely be excluded, the main significance for functional morphological studies has been found in the diameter of the SCs, whereas the radius of curvature is of minor interest. Additionally, we found clear differences in the bias angle of the canals between subterranean and gliding taxa, but also between sciurids and glirids. The sensitivity of the inner ear correlates with the locomotion mode, with a higher sensitivity of the SCs in fossorial species than in flying taxa. We conclude that the inner ear of flying and gliding mammals is less sensitive due to the large information flow into this sense organ during locomotion.

Background Feline carpal joint injuries pose a significant clinical challenge due to their complex anatomy and the limited evidence available to guide treatment decisions. A detailed understanding of the osseous microarchitecture is essential for improving surgical planning and therapeutic outcomes. A total of 80 carpal joints from 20 cats and 20 toy breed dogs were examined and compared using microcomputed tomography. The parameters bone volume (BV/TV), bone surface (BS/BV), trabecular thickness (Tb.Th), number of trabeculae (Tb.N), trabecular spacing (Tb.Sp), degree of anisotropy (DA) and connectivity density (Conn. D) were measured and compared. To classify the cortical structure, a three-staged scoring system was applied. Results All carpal bones exhibited a cancellous structure. Overall, cats had fewer but thicker trabeculae than the toy-dogs in the analysed areas of the carpus, which resulted in a greater trabecular distance in cats. Cats also had a higher bone volume (BV/TV) and thicker cortex. Conclusions There were significant differences in structure between dogs and cats which may be important when assessing carpal bone injuries.

Modern computational and imaging methods are revolutionizing the fields of comparative morphology, biomechanics, and ecomorphology. In particular, imaging tools such as X-ray micro computed tomography (μCT) and diffusible iodine-based contrast enhanced CT allow observing and measuring small and/or otherwise inaccessible anatomical structures, and creating highly accurate three-dimensional (3D) renditions that can be used in biomechanical modeling and tests of functional or evolutionary hypotheses. But, do the larger datasets generated through 3D digitization always confer greater power to uncover functional or evolutionary patterns, when compared with more traditional methodologies? And, if so, why? Here, we contrast the advantages and challenges of using data generated via (3D) CT methods versus more traditional (2D) approaches in the study of skull macroevolution and feeding functional morphology in bats. First, we test for the effect of dimensionality and landmark number on inferences of adaptive shifts during cranial evolution by contrasting results from 3D versus 2D geometric morphometric datasets of bat crania. We find sharp differences between results generated from the 3D versus some of the 2D datasets (xy, yz, ventral, and frontal), which appear to be primarily driven by the loss of critical dimensions of morphological variation rather than number of landmarks. Second, we examine differences in accuracy and precision among 2D and 3D predictive models of bite force by comparing three skull lever models that differ in the sources of skull and muscle anatomical data. We find that a 3D model that relies on skull μCT scans and muscle data partly derived from diceCT is slightly more accurate than models based on skull photographs or skull μCT and muscle data fully derived from dissections. However, the benefit of using the diceCT-informed model is modest given the effort it currently takes to virtually dissect muscles from CT scans. By contrasting traditional and modern tools, we illustrate when and why 3D datasets may be preferable over 2D data, and vice versa, and how different methodologies can complement each other in comparative analyses of morphological function and evolution.

Vertebrate models provide indispensable paradigms to study development and disease. Their analysis requires a quantitative morphometric study of the body, organs and tissues. This is often impeded by pigmentation and sample size. X-ray micro-computed tomography (micro-CT) allows high-resolution volumetric tissue analysis, largely independent of sample size and transparency to visual light. Importantly, micro-CT data are inherently quantitative. We report a complete pipeline of high-throughput 3D data acquisition and image analysis, including tissue preparation and contrast enhancement for micro-CT imaging down to cellular resolution, automated data processing and organ or tissue segmentation that is applicable to comparative 3D morphometrics of small vertebrates. Applied to medaka fish, we first create an annotated anatomical atlas of the entire body, including inner organs as a quantitative morphological description of an adult individual. This atlas serves as a reference model for comparative studies. Using isogenic medaka strains we show that comparative 3D morphometrics of individuals permits identification of quantitative strain-specific traits. Thus, our pipeline enables high resolution morphological analysis as a basis for genotype-phenotype association studies of complex genetic traits in vertebrates.

The increasing number of patients with end stage chronic kidney disease not only calls for novel therapeutics but also for pioneering research using convincing preclinical disease models and innovative analytical techniques. The aim of this study was to introduce a virtual histology approach using micro computed tomography (µCT) for the entire murine kidney in order to close the gap between single slice planar histology and a 3D high resolution dataset. An ex vivo staining protocol based on phosphotungstic acid diffusion was adapted to enhance renal soft tissue x-ray attenuation. Subsequent CT scans allowed (i) the detection of the renal cortex, medulla and pelvis in greater detail, (ii) the analysis of morphological alterations, (iii) the quantification of the volume as well as the radio-opacity of these portions and (iv) the quantification of renal fibrotic remodeling based on altered radio-opacity using the unilateral ureteral obstruction model. Thus, virtual histology based on PTA contrast enhanced CT will in future help to refine the outcome of preclinical research on kidney associated murine disease models.

Many bones are supported internally by a latticework of trabeculae. Scaling of whole bone length and diameter has been extensively investigated, but scaling of the trabecular network is not well characterized. We analysed trabecular geometry in the femora of 90 terrestrial mammalian and avian species with body masses ranging from 3 g to 3400 kg. We found that bone volume fraction does not scale substantially with animal size, while trabeculae in larger animals' femora are thicker, further apart and fewer per unit volume than in smaller animals. Finite element modelling indicates that trabecular scaling does not alter the bulk stiffness of trabecular bone, but does alter strain within trabeculae under equal applied loads. Allometry of bone's trabecular tissue may contribute to the skeleton's ability to withstand load, without incurring the physiological or mechanical costs of increasing bone mass.