The sequencing of ancient DNA has enabled the reconstruction of speciation, migration and admixture events for extinct taxa1. However, the irreversible post-mortem degradation2 of ancient DNA has so far limited its recovery-outside permafrost areas-to specimens that are not older than approximately 0.5 million years (Myr)3. By contrast, tandem mass spectrometry has enabled the sequencing of approximately 1.5-Myr-old collagen type I4, and suggested the presence of protein residues in fossils of the Cretaceous period5-although with limited phylogenetic use6. In the absence of molecular evidence, the speciation of several extinct species of the Early and Middle Pleistocene epoch remains contentious. Here we address the phylogenetic relationships of the Eurasian Rhinocerotidae of the Pleistocene epoch7-9, using the proteome of dental enamel from a Stephanorhinus tooth that is approximately 1.77-Myr old, recovered from the archaeological site of Dmanisi (South Caucasus, Georgia)10. Molecular phylogenetic analyses place this Stephanorhinus as a sister group to the clade formed by the woolly rhinoceros (Coelodonta antiquitatis) and Merck's rhinoceros (Stephanorhinus kirchbergensis). We show that Coelodonta evolved from an early Stephanorhinus lineage, and that this latter genus includes at least two distinct evolutionary lines. The genus Stephanorhinus is therefore currently paraphyletic, and its systematic revision is needed. We demonstrate that sequencing the proteome of Early Pleistocene dental enamel overcomes the limitations of phylogenetic inference based on ancient collagen or DNA. Our approach also provides additional information about the sex and taxonomic assignment of other specimens from Dmanisi. Our findings reveal that proteomic investigation of ancient dental enamel-which is the hardest tissue in vertebrates11, and is highly abundant in the fossil record-can push the reconstruction of molecular evolution further back into the Early Pleistocene epoch, beyond the currently known limits of ancient DNA preservation.
An adult female Sumatran rhinoceros was observed with a swelling in the left infraorbital region in March 2017. The swelling rapidly grew into a mass. A radiograph revealed a cystic radiolucent area in the left maxilla. In June 2017, the rhinoceros was euthanized. At necropsy, the infraorbital mass measured 21 cm × 30 cm. Samples of the infraorbital mass, left parotid gland, and left masseter muscle were collected for histopathology (Hematoxylin & Eosin, Von Kossa, Masson's trichrome, cytokeratin AE1/AE3, EMA, p53, and S-100). Numerous neoplastic epithelial cells showing pleomorphism and infiltration were observed. Islands of dentinoid material containing ghost cells and keratin pearls were observed with the aid of the two special histochemistry stains. Mitotic figures were rarely observed. All the neoplastic odontogenic cells and keratin pearls showed an intense positive stain for cytokeratin AE1/AE3, while some keratin pearls showed mild positive stains for S-100. All samples were negative for p53 and S-100 immunodetection. The mass was diagnosed as a dentinogenic ghost cell tumor.
Mirror-induced behaviour has been described as a cognitive ability of an animal to self-direct their image in front of the mirror. Most animals when exposed to a mirror responded with a social interactive behaviour such as aggressiveness, exploratory and repetitive behaviour. The objective of this study is to determine the mirror-induced self-directed behaviour on wildlife at the Royal Belum Rainforest, Malaysia. Wildlife species at the Royal Belum Rainforest were identified using a camera traps from pre-determined natural saltlick locations. Acrylic mirrors with steel frame were placed facing the two saltlicks (Sira Batu and Sira Tanah) and the camera traps with motion-detecting infrared sensor were placed at strategically hidden spot. The behavioural data of the animal response to the mirror were analysed using an ethogram procedure. Results showed that barking deer was the species showing the highest interaction in front of the mirror. Elephants displayed self-directed response through inspecting behaviour via usage of their trunk and legs while interacting to the mirror. Interestingly, the Malayan tapir showed startled behaviour during their interaction with the mirror. However, the absence of interactive behaviour of the Malayan tiger signalled a likelihood of a decreased social response behaviour. These results suggested that the ability to self-directed in front of the mirror is most likely related to the new approach to study the neural mechanism and its level of stimulus response in wildlife. In conclusion, research on mirror-induced self-directed behaviour in wildlife will have profound implications in understanding the cognitive ability of wildlife as an effort to enhance the management strategies and conservation.