The Upper Paleolithic hard animal tissue under the microscope: Selected examples from Moravian sites
Introduction
The histological approach is applied to hard tissues (bone, tooth, antler, and ivory) in forensic, archeological, paleoanthropological, and paleontological sciences. The Mid-Upper and Late Paleolithic material is usually fragmentary with marks of taphonomic changes and thus the taxonomic origin determination of the sample can be sometimes impossible on the macroscopic level of osteological research. Histometric and histomorphologic analyses are relevant for discerning the origin of human bones (Rämsch and Zerndt, 1963; Owsley et al., 1985; Cattaneo et al., 1999, 2009; Urbanová and Novotný, 2005; Hillier and Bell, 2007; Crescimanno et al., 2012; Cummaudo et al., 2019) and the taxonomic identity of animal species (Hidaka et al., 1998; Cuijpers, 2006; Martiniaková et al., 2006, 2007; Sawada et al., 2010, 2014; Padian and Lamm, 2013; Zhang et al., 2018). Observation of osteon banding and plexiform tissue is usually considered as the confirmation of non-human bone origin (Mulhern and Ubelaker, 2001, 2012; Cuijpers, 2006; Owsley et al., 1985; Muhlern and Ubelaker, 2012), although the presence of plexiform tissue was confirmed for human sub-adult individuals too (Hillier and Bell, 2007). Then the differences between individuals (Stout and Gehlert, 1979), including estimations of age at death, sex (Bouvier and Ubelaker, 1977; Eriksen, 1991; Thomas et al., 2000; Crowder and Stout, 2012), origin, and changes in pathological microstructures (Bancroft et al., 1996) can be addressed. Additionally, various biotic and abiotic taphonomic issues and questions discerning human intentional activities from other depositional and post-depositional processes can be raised (Binford, 1981; Hedges et al., 1995; Jans et al., 2002; Jans, 2005; Hanson and Cain, 2007; Hollund et al., 2011, 2018; Bell, 2012; Huisman et al., 2017; Brönnimann et al., 2018).
The periodical nature of hard dental tissue formation records (enamel and dentine) provides in histological sections critical information in species determination, age at death in the case of sub-adult individuals (Shellis, 1998; Dirks et al., 2002; Fitzgerald-Rose 2007), seasonality (Liebermann, 1993, 1994; Smith et al., 2006; Nývltová Fišáková, 2007, 2013; Dean, 2017), and survived stress events, such as malnutrition, infectious pathogens, trauma, weaning, or seasonal changes in resource availability (Macho et al., 1996; Dirks et al., 2002; Birch and Dean, 2014; Hupková et al., 2016; Hogg et al., 2018; Lorentz et al., 2019; Skinner and Byra, 2019). Furthermore, the periodicity of Retzius striae is variable between taxonomies and between individuals but individually is consistent (Shellis, 1998; Bromage et al., 2009; Hogg 2018). The periodical enamel formation can also provide information about the metabolism of an organism. According to the Havers-Halberg Oscillation (HHO) hypothesis, the long period growth marks visible in dental enamel as Retzius striae relate to the adult body mass and characteristic individual life history (Bromage et al., 2009, 2012; Mahoney et al., 2017). According to the HHO model, species with a shorter HHO periodicity, as expressed in the teeth as periodicity between Retzius striae, tend to have faster growth rates, smaller body size, and shorter lifespans (Hogg et al., 2018). However, the HHO model is not valid in the case of for example lemurs, when RP does not positively correlate with body mass (Hogg et al., 2015), the same for Canis familiaris (Hogg, 2018).
These methods provide framework for the study of individual micro life history, its ontogenetic development, and evolution of the species in the context of health, nourishment, seasonal strategies, environmental changes, and the depositional and post-depositional processes that affect hard tissue post-mortem. However, a major downside to these methods is the destructive process of sample manipulation. Thus, in cases where the osteological material is limited, as in paleoanthropology, electron microscopy or micro-CT analysis that provides virtual sectioning is preferred (Guatelli-Steinberg et al., 2014; Smith et al., 2018; Smith, 2019). Combining hard tissue sectioning with additional analytic methods such as isotopic or digital cementum luminescence analysis (e.g. Katzenberg and Lovell, 1999; Straight et al., 2004; Wall-Scheffler and Foley, 2008), for which precise sectioning is critical, could compensate the sample loss during the preparation process.
In our study, we present both non-metric and metric analyses of 17 burned and unburned animal bone samples and daily secretion rates and Retzius periodicity on three wolf teeth from selected Mid- and Late Upper Paleolithic Moravian sites. We describe the advantages and disadvantages of selected analytical micro-scale analyses to evaluate their potential for a) species determination of indeterminate burned and unburned fragments, b) variation in crown enamel mineralization, and c) micro-taphonomic changes that prevent the success of thin section analysis.
Section snippets
Material
In total, 20 fossil bone and tooth samples were analyzed in our paper (Table 1). The majority (15 instances + 1 without closer provenience) originated from the micro-region settlement area Dolní Věstonice I, II – Pavlov I, VI, and Milovice IV with a main cultural layer between 34 and 29 ka cal BP. The area presents a Gravettian site complex with hierarchical pattern of organization and various lengths of human occupation oriented on hunting of large herbivore herds (mammoths, reindeer, and
Methods
All samples selected for this study were anatomically described, measured, and photo-documented. Epoxy resin Araldite 2020 replicas were prepared for all teeth before sectioning. Calcified sections of hard tissues were prepared according to already published protocols (Chinsamy and Raath, 1992; Bancroft et al., 1996; Macho et al., 1996; Beynon et al., 1998; Reid et al., 1998; Paral et al., 2007; Hogg, 2010; Padian and Lamm, 2013; García-Donas et al., 2017) in the Histological Laboratory at the
Burned and unburned bones and their origin
In 12 bone samples which showed signs of burning at the macroscopic level, three degrees of burning were observed and analyzed: a) low to medium degree of burning (samples bDVII-A, bDVII-B, bDVII-C, and bMIV-C) with brownish matrix, no cracking, and minimal deposition of carbon; b) medium to the high degree of burning (bDVI-A, bDVI-B, bDVI-C, bDVI-D, bMIV-B, and bPVI-A) with dark brown matrix and cracks spreading from and/or through the Haversian canals; and c) high to extremely high degree of
Discussion
Bone and tooth sample selection is the most important step in the histological analysis (Padian and Lamm, 2013). For example, when age, species determination, and ontogeny are analyzed, midshaft sections from long limb bones such as femur, tibia, humerus, or radius are preferred (Bouvier and Ubelaker, 1977; Parfitt et al., 1987; Urbanová and Novotný, 2005; Cuijpers, 2006; Martiniaková et al., 2006, 2007; Kolb et al., 2015; Cummaudo et al., 2019). Uniformity in section area eliminates the
Conclusion
We present comprehensive non-metric and metric analyses of 17 burned and unburned animal bone fragments as well as daily secretion rates and Retzius periodicity on three wolf premolars from selected Mid- and Late Upper Paleolithic sites from Moravia (Czech Republic). Non-metric characteristics of osteon banding and plexiform bone were confirmed in a sample (bSSIV-B) from Stránská skála IV. The metric characteristics supported its non-human origin and the characteristics suggest a medium-sized
Declaration of competing interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgments
This work was supported by the Czech Science Foundation [grant number: 20-26094S] and Czech national institutional support at Czech Academy of Sciences [grant number: RVO:68081758], both fundings awarded to the Institute of Archeology in Brno, Czech Academy of Sciences. We thank Nikol Dunajová and Eliška Ambrosová from the Open Science Programme at the Czech Academy of Sciences for help with Milovice IV and Dolní Věstonice I sample preparation.
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