Bacteria play an important role in the fossilization of soft tissues; their metabolic activities drive the destruction of the tissues and also strongly influence mineralization. Some environmental conditions, such as anoxia, cold temperatures, and high salinity, are considered widely to promote fossilization by modulating bacterial activity. However, bacteria are extremely diverse, and have developed metabolic adaptations to a wide range of stressful conditions. Therefore, the influence of the environment on bacterial activity, and of their metabolic activity on fossilization, is complex. A number of examples illustrate that simple, general assumptions about the role of bacteria in soft tissue fossilization cannot explain all preservational pathways: (i) experimental results show that soft tissues of cnidaria decay less in oxic than anoxic conditions, and in the fossil record are found more commonly in fossil sites deposited under oxic conditions rather than anoxic environments; (ii) siderite concretions, which often entomb soft tissue fossils, precipitate due to a complex mixture of sulfate‐ and iron reduction by some bacterial species, running counter to original theories that iron reduction is the primary driver of siderite concretion growth; (iii) arthropod brains, now widely accepted to be preserved in many Cambrian fossil sites, are one of the first structures to decay in taphonomic experiments, indicating that their fossilization processes are complex and influenced by bacterial activity. In order to expand our understanding of the complex process of bacterially driven soft tissue fossilization, more research needs to be done, on fossils themselves and in taphonomic experiments, to determine how the complex variation in microbial metabolic activity influences decay and mineralization.

Experiments are reported to reconstruct the taphonomic pathways of fish toward fossilisation. Acrylic glass autoclaves were designed that allow experiments to be carried out at elevated pressure up to 11 bar, corresponding to water depths of 110 m. Parameters controlled or monitored during decay reactions are pressure, salinity, proton activities (pH), electrochemical potentials (Eh), and bacterial populations. The most effective environmental parameters to delay or prevent putrefaction before a fish carcass is embedded in sediment are (1) a hydrostatic pressure in the water column high enough that a fish carcass may sink to the bottom sediment, (2) hypersaline conditions well above seawater salinity, and (3) a high pH to suppress the reproduction rate of bacteria. Anoxia, commonly assumed to be the key parameter for excellent preservation, is important in keeping the bottom sediment clear of scavengers but it does not seem to slow down or prevent putrefaction. We apply our results to the world-famous Konservat-Lagerstätten Eichstätt-Solnhofen, Green River, and Messel where fish are prominent fossils, and reconstruct from the sedimentary records the environmental conditions that may have promoted preservation. For Eichstätt-Solnhofen an essential factor may have been hypersaline conditions. Waters of the Green River lakes were at times highly alkaline and hypersaline because the lake stratigraphy includes horizons rich in sodium carbonate and halite. In the Messel lake sediments some fossiliferous horizons are rich in FeCO 3 siderite, a mineral indicating highly reduced conditions and a high pH. Since the advent of experimental methods in paeontological research, our understanding of taphonomic and fossilisation reactions has much improved. Today we realise how easily and rapidly organic tissue may be transformed into inorganic materials 1-6. Consensus is emerging that fossilisation reactions can take place within time frames accessible with laboratory experiments 7. The near-perfect articulation of the Pycnodontid in Fig. 1 suggests that the decision for or against preservation must have been made early, shortly after the fish died. Under ambient marine conditions-oxygenated water, normal marine salinity, and near-neutral pH-a fish so delicate would have been disarticulated or consumed by scavengers within hours to days. But what are the environmental factors most effective in retarding or preventing organic decay? If we identify those variables experimentally, we may hold the key to understanding the genesis of Konservat-Lagerstätten within which fish are prominent fossils. We report novel decay experiments with fish to understand early taphonomic pathways toward fossilisation. Parameters investigated experimentally are elevated hydrostatic pressure, elevated salinity, the role of proton (pH) and electron activity (Eh), bacterial activity, and time. We apply our results to three prominent Konservat-Lagerstätten where fossil fish are prominent species-Eichstätt-Solnhofen, Green River, and Messel. Previous taphon...

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The fossilization of soft tissues is generally the replacement of organic structures by pseudomorphs in which muscle tissue is mostly replaced by minerals (i.e., phosphate, carbonate or pyrite). Micro-CT observations of decomposing crayfish in tank and distilled water, show a precipitation of crystal clusters over time. In addition, a mineralized muscle was found by SEM analyses. Raman spectroscopy (CRS) revealed that crystal clusters and the muscle consist of well-ordered calcite. Inductively coupled plasma mass spectrometry (ICPMS) of the distilled water showed a calcium content below the detection limit at the beginning of the experiments, which indicates that most of the calcium ions needed for the precipitation were provided by the decomposing carcasses themselves. Volume measurements of 3D-reconstructed calcite clusters and gastroliths showed a general increase of the volume of calcite clusters and simultaneously volume reduction of gastroliths with progressive decay. Specimens that were in the postmoult phase showed a smaller total volume of precipitated calcite, compared to specimens, which were in the intermoult or premoult phase. In addition, measurements of the total amount of body calcium of Cambarellus diminutus by atomic absorption spectrophotometry (AAS) revealed a higher amount of calcium in individuals without gastroliths than in individuals with gastroliths. It is assumed, that the higher the body size, the higher the volume of precipitated calcite, if the individuals were in the intermoult phase at the time of death. If the individuals were in the postmoult or premoult phase, the phase itself seems to be important.

Fossilization processes and especially the role of bacterial activity during the preservation of organic material has not yet been well understood. Here, we report the results of controlled taphonomic experiments with crayfish in freshwater and sediment. 16S rRNA amplicon analyzes showed that the development of the bacterial community composition over time was correlated with different stages of decay and preservation. Three dominating genera, Aeromonas, Clostridium and Acetobacteroides were identified as the main drivers in the decomposition of crayfish in freshwater. Using micro-computed tomography (µ-CT), scanning electron microscopy (SEM) and confocal Raman spectroscopy (CRS), calcite clusters were detected after 3–4 days inside crayfish carcasses during their decomposition in freshwater at 24 °C. The precipitation of calcite clusters during the decomposition process was increased in the presence of the bacterial genus Proteocatella. Consequently, Proteocatella might be one of the bacterial genera responsible for fossilization.

The preservation of soft tissue in the fossil record is mostly due to the replacement of organic structures by minerals (e.g. calcite, aragonite or apatite) called pseudomorphs. In rare cases soft tissues were preserved by pyrite. We assume that adipocere, as the shaping component, might be a preliminary stage in the pyritisation of soft tissues under anaerobic conditions. Using high-performance liquid chromatography coupled to ultraviolet and mass spectrometric detection (HPLC–UV/MS) and confocal Raman spectroscopy (CRS) we were able to demonstrate the transformation of the hepatopancreas (digestive gland) of the crayfish Cambarellus diminutus [Hobbs 1945] into adipocere within only 9 days, just inside a biofilm. Microorganisms (bacteria and fungi) which were responsible for the biofilm (Sphaerotilus [Kutzig 1833] and Pluteus [Fries 1857]) and maybe the adipocere formation (Clostridium [Prazmowski 1880]) were detected by 16S rRNA gene amplicon sequencing. Furthermore, micro-computed tomography (µ-CT) analyses revealed a precipitation of calcite and further showed that in animals with biofilm formation calcite precipitates in finer grained crystals than in individuals without biofilm formation, and that the precipitates were denser and replicated the structures of the cuticles better than the coarse precipitates.

Past claims have been made for fossil DNA recovery from various organisms (bacteria, plants, insects and mammals, including humans) dating back in time from thousands to several million years BP. However, many of these recoveries, especially those described from million-year-old amber (fossil resin), have faced criticism as being the result of modern environmental contamination and for lack of reproducibility. Using modern genomic techniques, DNA can be obtained with confidence from a variety of substrates (e.g. bones, teeth, gum, museum specimens and fossil insects) of different ages, albeit always less than one million years BP, and results can also be obtained from much older materials using palaeoproteomics. Nevertheless, new attempts to determine if ancient DNA (aDNA) is present in insects preserved in 40 000-year old sub-fossilised resin, the precursor of amber, have been unsuccessful or not well documented. Resin-embedded specimens are therefore regarded as unsuitable for genetic studies. However, we demonstrate here, for the first time, that although a labile molecule, DNA is still present in platypodine beetles (Coleoptera: Curculionidae) embedded in six-year-old and two-year-old resin pieces from Hymenaea verrucosa (Angiospermae: Fabaceae) collected in Madagascar. We describe an optimised method which meets all the requirements and precautions for aDNA experiments for our purpose: to explore the DNA preservation limits in resin. Our objective is far from starting an uncontrolled search for aDNA in amber as it was in the past, but to start resolving basic aspects from the DNA preservation in resin and search from the most modern samples to the ancient ones, step by step. We conclude that it is therefore possible to study genomics from resinembedded organisms, although the time limits remain to be determined.

Premise Within a broader study on leaf fossilization in freshwater environments, a long‐term study on the development and microbiome composition of biofilms on the foliage of aquatic plants has been initiated to understand how microbes and biofilms contribute to leaf decay and preservation. Here, water lily leaves are employed as a study model to investigate the relationship between bacterial microbiomes, biodegradation, and fossilization. We compare four DNA extraction kits to reduce biases in interpretation and to identify the most suitable kit for the extraction of DNA from bacteria associated with biofilms on decaying water lily leaves for 16S rRNA amplicon analysis. Methods We extracted surface‐associated DNA from Nymphaea leaves in early stages of decay at two water depth levels using four commercially available kits to identify the most suitable protocol for bacterial extraction, applying a mock microbial community standard to enable a reliable comparison of the kits. Results Kit 4, the FastDNA Spin Kit for Soil, resulted in high DNA concentrations with better quality and yielded the most accurate depiction of the mock community. Comparison of the leaves at two water depths showed no significant differences in community composition. Discussion The success of Kit 4 may be attributed to its use of bead beating with a homogenizer, which was more efficient in the lysis of Gram‐positive bacteria than the manual vortexing protocols used by the other kits. Our results show that microbial composition on leaves during early decay remains comparable and may change only in later stages of decomposition.