Biomolecules in Pleistocene fossils from tropical cave indicate fossil biofilm

Finding biomolecules in fossils is a challenging task due to their degradation over time from physical, chemical, and biological factors. The primary hypothesis for explaining the presence of biomolecules in fossilized bones tissues suggests their survival in the fossilization process. In contrast, some of these biomolecules could either derive from bacteria biofilm, thus without a direct relationship with the fossil record or could be an artifact from measurement procedures. Raman spectroscopy studies across various fossil ages and environments have detected multiple bands ranging from 1200 to 1800 cm^-1 associative of organic compounds. However, the significance of these bands remains elusive. Our research aims to address this issue through a deep Raman spectroscopy investigation on Pleistocene teeth from Tayassu and Smilodon populator. These fossils were obtained from a well-preserved stratigraphic succession in Toca de Cima do Pilão cave, near the National Park of Serra da Capivara in semiarid Brazil. We propose two hypotheses to explain the presence of organic compounds related to 1200 to 1800 cm^-1 Raman spectral range in fossil tissues: (i) these bands are biological signatures of preserved fossil biomolecules, or (ii) they are exogenous biological signatures associated with the bacterial biofilm formation during post-depositional processes. Our results align with the latter hypothesis, followed by biofilm degradation. However, the specific mechanisms involved in the natural biofilm degradation in fossil records remain unexplored in this study. In our case, the formation of biofilm on fossil bones is attributed to the oligotrophic conditions of the cave sediment matrix. We present a comprehensive model to elucidate the existence of biofilm on fossilized tissues, emphasizing the pivotal role of post-depositional processes, especially water action, in the cave environment. As the fossils were discovered in a cave setting, post-depositional processes significantly contribute to the formation of the biofilm matrix. Although our study provides insights into biofilm formation, further research is needed to delve into the specific mechanisms driving natural biofilm degradation in fossils.