Unique bone histology of modern giant salamanders: a study on humeri and femora of Andrias spp.

The osteohistology of Andrias spp. is a pivotal analogue for large fossil non-amniotes (e.g., Temnospondyli), and the endangered status of this taxon underlines the importance of gathering information on its growth. We here present the first osteohistological study by petrographic thin sections of an ontogenetic series of humeri and femora of eight individuals of varying sizes (28.5-104 cm) and ages (2.5-32 years) of Andrias japonicus from the Hiroshima City Asa Zoological Park, Japan. In addition, two individuals of A. cf. davidianus of unknown age but of different size (62 cm and 94 cm) were studied. All samples of Andrias spp. show a primary avascular periosteal cortex made of parallel-fibred tissue around the ossification center in the petrographic thin sections. Mainly in small individuals, the fibers forming this tissue are very coarse and loosely organized. With increasing size and age, the coarse tissue is irregularly intermixed and later replaced with finer and better organized fibers. This histologic change is accompanied by a change from diffuse annuli in the inner cortex to distinct lines of arrested growth (LAGs) in the outer cortex. We interpret these changes in tissue and the appearance of distinct growth marks as indicating the onset of active reproduction. The lack of primary vascularization around the ossification center in our Andrias spp. sample is striking and contradicts other observations. Vascularity may be prone to plasticity and further studies are necessary. We hypothesize that the large osteocyte lacunae and the dense networks of canaliculi observed in our sample may have nourished the tissue instead of primary vascular canals. We measured the size of osteocyte lacunae of Andrias spp. in comparison to other Lissamphibia, and found them to be significantly larger throughout ontogeny. The periosteal cortex contains a high amount of thick Sharpey's fibers all around the midshaft cross sections. The two samples of Andrias cf. davidianus show tissue and growth mark distribution similar to that observed in A. japonicus. However, the large individual of A. cf. davidianus differed in its extremely osteosclerotic condition and the retention of a small layer of calcified cartilage in the endosteal region of the femur. It remains unclear whether these differences are related to plasticity, taxonomy, sex, exogenous factors, or attributable to a regenerated but fully regrown leg. Although the present study is based on zoo-kept and not wild, animals, it yields important insights into osteohistological plasticity and growth patterns in giant salamanders.