A multidisciplinary approach to reconstruct Upper Palaeolithic and Mesolithic dietary habits:

Human adaptation to Pleistocene-Holocene environmental change in northeastern Italy

Figure 2. Ternary diagrams. Relation between Villabruna, Tagliente, Mondeval and (a) Modern human groups

composed by Vancouver Island (gray), Inuit (green) and Khoen-San (yellow) and (b) Fossils comparison sample

composed by Homo neanderthalensis (blue), Early Homo sapiens (Orange) and Natufians (red).

In this work, we investigated different aspects of the cultural and dietary

adaptations of three hunter-gatherer male individuals who were buried

between Late Pleistocene and the Early Holocene in three sites of north-

eastern Italy of Riparo Tagliente (Verona), Riparo Villabruna (Belluno) and

Mondeval de Sora (Belluno).

Figure 1. (a) Bone collagen stable isotope values in human remains from Upper Paleolithic sites of

Italy (b) Differences between human and herbivore δ15N (‰) of bone collagen.

While the distribution of dental macrowear can be probably related to a number of variables

affecting the oral cavity such as diet (Fiorenza et al. 2011) and para-functional asymmetries

(Oxilia et al. 2018), our results suggest that its use as the only reliable proxy for dietary

habits is statistically supported when differences between groups are extreme. Isotope results

indicate that animal proteins and freshwater resources were likely consumed by Upper

Palaeolithic and Mesolithic individuals of north-eastern Italy together. Residues may have

been left by para-masticatory activities. In conclusion, we highlight the importance of using

different analytical methods to overcome the limitations imposed by a single method. A

multi-proxy approach can shed light on the stratified traces left by both dietary and non-

dietary habits in human fossils.

Sta

ble

Isoto

pes

Gregorio Oxilia1,2, Federica Fontana3, Luca Fiorenza4,5, Ottmar Kullmer6,7, Gwenaëlle Goude8, Valentina Gazzoni3, Federico Lugli2,9, Marco Peresani3,

Matteo Romandini2,3, Elisabetta Cilli2, Claudio Tuniz10,11,12, Federico Bernardini11,10,, Eugenio Bortolini2, Jessica C. Menghi Sartorio2, Sahra Talamo13,

Stefano Benazzi2,13, Emanuela Cristiani1.

1 - Department of Oral and Maxillo Facial Sciences Sapienza University, Via Caserta, 6 00161, Rome, Italy · 2 - Department of Cultural Heritage University of Bologna Via degli Ariani 1 48121 Ravenna, Italy · 3 - Dipartimento di Studi Umanistici – Sezione di Scienze

Preistoriche e Antropologiche, Università degli Studi di Ferrara, Corso Ercole I d’Este 32 · 4 - Department of Anatomy and Developmental Biology, Monash University, Melbourne VIC 3800, Australia · 5 - Earth Sciences, University of New England, Armidale NSW 2351,

Australia · 6 - Senckenberg Research Institute, Senckenberganlage 25, 60325 Frankfurt am Main, Germany · 7 - Department of Paleobiology and Environment, Institute of Ecology, Evolution, and Diversity, Johann Wolfgang · 8 - Aix-Marseille Université, CNRS, MCC UMR

7269 - Laboratoire Méditerranéen de Préhistoire Europe Afrique Maison Méditerranéenne des Sciences de l’Homme 5 rue du Château de l’Horloge - B.P. 647 13094 · 9 - Department of Chemical and Geological Sciences, University of Modena and Reggio Emilia, Italy · 10 -

The “Abdus Salam” International Centre for Theoretical Physics, Trieste, Italy ·11 - Museo Storico della Fisica e Centro Studi e Ricerche "Enrico Fermi", Rome, Italy · 12 - Center for Archaeological Science, University of Wollongong, Australia · 13 - Department of Human

Evolution, Max Planck Institute for Evolutionary Anthropology

References

Vercellotti, G., Alciati, G., Richards, M. P., & Formicola, V. (2008). The Late Upper Paleolithic skeleton Villabruna 1 (Italy): a

source of data on biology and behavior of a 14.000 year-old hunter. Journal of Anthropolological Sciences, 86, 143-163.

Gazzoni, V., Goude, G., Herrscher, E., Guerreschi, A., Antonioli, F., & Fontana, F. (2013). Late Upper Palaeolithic human diet: first

stable isotope evidence from Riparo Tagliente (Verona, Italy). Bulletins et Mémoires de la Société d'Anthropologie de Paris, 25(3-

4), 103-117.

Fiorenza, L., Benazzi, S., Tausch, J., Kullmer, O., Bromage, T.G., & Schrenk, F. (2011). Molar macrowear reveals Neanderthal eco-

geographic dietary variation. PLoS ONE, 6, e14769.

Aim

We combined Dental Calculus, Macrowear Pattern and Stable Isotope Analyses on the

three individuals. Macrowear analysis was conducted on lower M2s in all the individuals as

well as on our reference sample, which included fossils from the Middle and Upper

Palaeolithic (PALAEO, n = 29), Natufians (NAT, n = 8), and recent populations with known

diet: Inuit (n = 4), Vancouver Islanders (n = 3), and Khoe‐San (n = 4).

Pairwise Euclidean distance was computed among all individuals considering the three

masticatory phases (Phase II, Buccal, Lingual). The relevance of each categorical variable

(Human group, Geography, Culture, Chronology) was then tested through AMOVA.

Additionally, the presence of differences among variables was inspected by Kruskal-Wallis

test. Pairwise differences were explored through Mann-Whitney U test. The relationship

between Villabruna, Tagliente and Mondeval samples and the reference groups was formally

assessed via a t-test and inspected by computing Euclidean and Mahalanobis distance

between the samples and each reference group’s centroid.

Materials and Methods

Results

Conclusions

This study was supported by the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation

programme (agreement no.639286, HIDDEN FOODS www.hiddenfoods.eu). We also thanks grant agreement No 724046 – SUCCESS

http://www.erc-success.eu/.

Acknowledgements

All the studied individuals show δ15N bone collagen values well above a trophic step shift

(3-5‰) when compared to the local fauna. This indicates that their diet was largely based

on terrestrial animal proteins rather than plant foods (Vercellotti et al., 2008; Gazzoni et

al., 2013). For all these individuals we cannot exclude the additional intake of aquatic

resources. In particular, a possible explanation for the relatively high δ13C and δ15N

values of one of the Tagliente individuals is the consumption of marine and/or freshwater

resources (ca. 20% of the total protein intake).

Den

talca

lculu

s

Micro-fossils entrapped in dental calculus were particularly abundant on the individual

found at Mondeval de Sora. Here, the consumption of different plant foods is suggested

on the basis of the recovery of different types of starch granules, which are consistent in

size and shape with those of the Poaceae grass family and specifically with the tribes

Avenae and Triticeae. The presence of non-dietary micro-residues such as plant fibres and

wood particles in the dental calculus suggests that this individual used mouth and teeth in

extra-masticatory activities (Alciati et al. 1995). Fragments of feathers were also

identified with shape and features at their nodes that would be consistent with aquatic

birds of the Anatidae family.

Macr

ow

ear

Dental wear patterns of the three individuals are concentrated in the Lingual area

(Figure 2) and consistent with a plant-based diet (Fiorenza et al. 2011). Exploratory

analyses, however, found significant differences only for Buccal phase I between

Natufians and Neanderthal (W=145.5, p=0.0029 ; corrected p-value=0.043), as

well as between Middle and Late Upper Palaeolithic individuals (W=170.5,

p=0.0032; corrected p-value=0.019). Considering the relationship between the

three samples of interest (Villabruna, Tagliente and Mondeval) and reference

groups, significant differences were found between Villabruna and Khoe San for

Phase II (t=3.45, p=0.02) and Buccal Phase I (t=-4.22, p=0.012) (Figure 2b and 3).

Figure 3. A hierarchical clustering. NAT = Natufians, UPHS = Upper Palaeolithic Homo sapiens, EHS

= Early Homo sapiens, HS_Hunt = Inuit/Vancouver Island, HS_mixed = Khoe San

Corrisponding author: gregorio.oxilia@uniroma1.it

Figure 2. Microfossils entrapped in dental calculus from Mondeval: (a) woody debris; (b and c)

starch with bimodal distribution; (d) fragment of feather.

(b) (c) (a)

20μm20μm 10μm20μm

(b) (c) (d)

Oxilia, G., Bortolini, E., Martini, S., Papini, A., Boggioni, M., Buti, L., Figus, C., Sorrentino, R., Townsend, G., Kaidonis, J., Fiorenza,

L., Cristiani, E., Kullmer, O., Moggi‐Cecchi, J., Benazzi, S. (2018). The physiological linkage between molar inclination and dental

macrowear pattern. Am J Phys Anthropol.,166:941–951.

Alciati G., Coppa, Macchiarelli R. 1995. La dentizione del cacciatore mesolitico di Mondeval de Sora (San Vito di Cadore,

Belluno). Bullettino di Paletnologia Italiana, 86, 153-196.