Microsc. Microanal. 23, 1207–1213, 2017 doi:10.1017/S1431927617012661


MICROGRAPHIA © MICROSCOPY SOCIETY OF AMERICA 2017


The Application of Scanning Electron Microscopy with Energy-Dispersive X-Ray Spectroscopy (SEM-EDX) in Ancient Dental Calculus for the Reconstruction of Human Habits


Dana Fialová,1,* Radim Skoupý,2 Eva Drozdová,1 Aleš Paták,2 Jakub Piňos,2 Lukᚊín,3 Radoslav Beňuš,4 and Bohuslav Klíma5


1Department of Experimental Biology, Faculty of Science, Masaryk University, Kamenice 5, 625 00 Brno, Czech Republic 2Institute of Scientific Instruments of the CAS, Královopolská 147, 612 64 Brno, Czech Republic 3Archaeological Centre Olomouc, U Hradiska 6, 779 00 Olomouc, Czech Republic 4Department of Anthropology, Faculty of Natural Sciences, Comenius University, Ilkovičova 6, 842 15 Bratislava 4,


Slovak Republic 5Department of History, Faculty of Education, Masaryk University, Poříčí 9, 603 00 Brno, Czech Republic


Abstract: The great potential of scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM-EDX) is in detection of unusual chemical elements included in ancient human dental calculus to verify hypotheses about life and burial habits of historic populations and individuals. Elemental spectra were performed from archeological samples of three chosen individuals from different time periods. The unusual presence of magnesium, aluminum, and silicon in the first sample could confirmthe hypothesis of high degree of dental abrasion caused by particles from grinding stones in flour. In the second sample, presence of copper could confirm that bronze jewelery could lie near the buried body. The elemental composition of the third sample with the presence of lead and copper confirms the origin of individual to NapoleonicWars because the damage to his teeth could be explained by the systematic utilization of the teeth for the opening of paper cartridges (a charge with a dose of gunpowder and a bullet),whichwere used during the 18th and the 19th centuryAD.All these results contribute to the reconstruction of life (first and third individual) and burial (second individual) habits of historic populations and individuals.


Key words: ancient dental calculus, SEM-EDX, human habits, the Great Moravian Empire, Napoleonic Wars INTRODUCTION


tified as a reservoir of various fragments of cereals, vegetable fibers, phytoliths, pollens, seeds, animal hairs, parasites, insects, microfossils, bacteria, fungi, other biomolecules and elements. Several different types of microscopes were used to examine ancient human dental calculus. Polarizing light microscopes were used for identification of microfossils such as phytoliths or starch grains, which are important in revealing the human diet (Boyadjian et al., 2007; Piperno & Henry, 2008; Hardy et al., 2009, 2012). Transmission electron microscopy (TEM) brought information about bacterial components, crystals (Kakei et al., 2009) and also proof that “reactive” bacterial ancient DNA (aDNA) exists in 4,000–5,000 years old human dental calculus (Preus et al., 2011). Scanning electron microscopy (SEM) gave a lot of information about human diet (Arensburg, 1996;


Ancient human dental calculus is a very important bio-archeological material because it is formed from dental plaque during the life of an individual. Thus, it has been iden-


*Corresponding author.danafialka@mail.muni.cz Received February 14, 2017; accepted October 18, 2017


Fox et al., 1996; Hardy et al., 2012; Power et al., 2014), oral bacterial flora(Vandermeerschetal.,1994; Papetal.,1995; Linossier et al., 1996; Hershkovitz et al., 1997), environmental conditions (Fox et al., 1996), and the habits of historic popu- lations and individuals (Blatt et al., 2011; Hardy et al., 2012). Energy-dispersive X-ray spectroscopy (EDX) was used


in connection with TEM and brought information about the inorganic elemental composition of the calculus of recent ethnic groups. Differences between two ethnic groups were shown mainly in the levels of sodium and magnesium and in relation to smoking (Roberts-Harry et al., 2000). The inorganic and crystallographic composition of recent calculus has been known for many years (Gron et al., 1967; Sundberg & Friskopp, 1985). The main elements in dental calculus are calcium, phosphorus, magnesium, fluoride, carbon dioxide and they mainly come from hydroxyapatite, whitlockite, octacalciumphosphate, and brushite (Gron et al., 1967).Other special elements are supposed to come from other sources dependent on diet, habits, environment, ethnic groups, etc. SEM-EDX was also used in connection with SEM to


focus on microfossils and starch in ancient dental calculus (Dudgeon & Tromp, 2014; Power et al., 2014). Only the


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