First Scanning electron microscope analysis of dental calculus from European Neanderthals: Subalyuk, (Middle Paleolithic, Hungary). Preliminary report

Ancient human microbiomes

“…Dental calculus is a calcified bacterial biofilm that forms on the surfaces of teeth, and it is found in all human populations, as well as Miocene apes (12.5–8.5 Ma) (Hershkovitz et al, 1997), Neanderthals (Pap et al, 1995; Henry et al, 2011; Hardy et al, 2012), wild chimpanzees (Hardy et al, 2009), and a range of animals (Dobney and Brothwell, 1987; Middleton and Rovner, 1994). Among humans, both in the past and today when professional dentistry care is not available, the incidence of dental calculus is near-ubiquitous among adults by age 30 (White, 1997; Lieverse, 1999), and in our experience we have found that it is not uncommon to observe dental calculus deposits in excess of 100 milligrams in archaeological assemblages of agricultural populations.…”

“…Following electron microscopy imaging of modern dental calculus in the 1960s and 1970s (Schroeder, 1969; Jones, 1972; Lustmann et al, 1976), it was recognized that microorganisms within human, Neanderthal, and extinct primate dental calculus could be imaged and morphologically characterized using SEM (Brothwell, 1972; Dobney and Brothwell, 1986, 1988; Hansen et al, 1991; Dobney, 1994; Vandermeersch et al, 1994; Pap et al, 1995; Arensburg, 1996; Hershkovitz et al, 1997) and later direct optical techniques (Linossier et al, 1996; Charlier et al, 2010). Biomolecular investigations of calculus began with immunohistochemical analysis of Streptococcus mutans (Linossier et al, 1996), followed by gold-labeled antibody transmission electron microscopy (TEM) of in situ DNA (Preus et al, 2011), and PCR-based analyses targeting specific oral taxa, including Actinomyces naeslundii, Fusobacterium nucleatum, Streptococcus gordonii, P. gingivalis and S. mutans (De La Fuente et al, 2012; Adler et al, 2013).…”

Bacteria and archaea paleomicrobiology of the dental calculus: a review

“…Among 27–85% of affected teeth, dental calculus was more frequently observed on the buccal side of the maxillary first molars and on the lingual side of the mandibular incisors (Whittaker et al ., ; Bonfiglioli et al ., ; Belcastro et al ., ; Flensborg, ; Vodanović et al ., ; Masotti et al ., ). Microscopic observations evidenced the omnipresence of well‐preserved calcified microorganisms in ancient human calculus of modern, Neolithic, Epipaleolithic, and Middle Paleolithic populations from 60,000 years before present ( bp ) by using optical microscopy after Gram staining (Charlier et al ., ; Warinner et al ., ,b), scanning electron microscopy (Dobney & Brothwell, ; Dobney, ; Pap et al ., ; Arensburg, ; Meller et al ., ; Charlier et al ., ; Warinner et al ., ,b), transmission electron microscopy (Preus et al ., ), fluorescence microscopy with DNA fluorescent dye to reveal dsDNA (Warinner et al ., ,b), fluorescence in situ hybridization to reveal archaea (Huynh et al ., unpublished data ) and immunohistochemical analysis incorporating polyclonal antibody specific to Streptococcus mutans (Linossier et al ., ). Scanning electron microscopy revealed the progressive build‐up of ancient dental calculus with remarkable microscopic incremental growth lines (Adler et al ., ).…”

Comments on Soltysiak's paper: “Comment: Low dental caries rate in Neandertals: The result of diet or the oral flora compositions?”

“…Do et al, 2012). However, in dental calculus from Middle Palaeolithic skeletons (Kebara 2 and Subalyuk 1) these bacteria were identified (Arensburg, 1996;Pap et al, 1995;Vandermeersch et al, 1994). S. mutans is usually assigned the largest role in the formation of dental caries.…”