prior to this study and all genetic data from the coprolite were obtained prior to the analyses of modern hyena samples, which were performed later in the modern DNA laboratory.

The contamination of the coprolite may have occurred in South Africa, for the Azokh coprolite was sawed for pollen analysis in the same laboratory room in Bloemfontein in which 10 days before fresh brown hyena scats had been cleaned. Thus, in spite of careful preparation to avoid pollen contamination, residues might have contaminated the coprolite before it was returned to the sample bag immediately after sawing. We believe therefore that the most likely explanation for the presence of brown hyena DNA sequences in the Azokh coprolite is that it had been contaminated with modern brown hyena DNA through secondary contact (bench surface, saw etc.) or residues produced in the Bloemfontein laboratory. This explanation is more parsimonious than would be the reappraisal of past brown hyena distribution with a range extending up to NagornoKarabakh. The fact that, apart from brown hyena, no other carnivore sequences were obtained via the targeted PCR approach, and that no indication of the producer’s species was found in the genomic data set, taken together with our previous investigations of numerous cave bear bones from the Azokh cave from which no PCR product was obtained, argue in favor of poor endogenous DNA preservation in the fossil remains of the Azokh cave. Samples with poor DNA preservation, however, are particularly prone to produce artifactual results in paleogenetic studies. The present study highlights the importance of addressing the problems of contamination starting at the very early stages of sample collection during field work when a paleogenetic analysis of the samples is considered. Taken together with our previous demonstration of the importance of early sample treatment to favor optimal DNA preservation (Pruvost et al. 2007) our work reveals the importance of a close collaboration between molecular geneticists and archaeologists or paleontologists.

Conclusions

1.Two coprolites recovered from Azokh 1, Unit II have been studied.

2.Their size and form are comparable to hyena scats, but there is no indication in the form of bone crushing or tooth marks that hyenas were present. No hyena fossils have been recovered from Unit II so far, but they are known in underlying deposits in Unit V.

3.The most abundant species in this site is the cave bear (Ursus spelaeus), an extinct species whose dietary and living behaviors have been considered to be different (though still controversial) to modern bears.

4.The much larger body size of U. spelaeus compared to the largest sized hyena recorded in the site, should have produced larger sized coprolites. Indeed, comparisons with hyena coprolites from other fossil sites show that bear coprolites are larger than the Azokh coprolites. Coprolite morphomotry has not been conclusive. However, the involvement of hyenas with no further taxonomic remains or taphonomic evidence of their presence, except for their coprolites, appears dubious.

5.Chemical analyses of the coprolites by diffraction and fluorescence suggest the possibility of hyenas as the

coprolite producer by the presence of the bone mineral (hydroxylapatite). However, hydroxyapatite is the most common and stable neo-formed mineral derived from bat guano diagenesis, which is very intense in Unit II, and has actually been identified in geological materials of Unit II, such as stones and sediments.

6.Relatively high content of amorphous phases and low crystallinity in both geological (stones and sediments)

and biological (coprolites) samples may agree with neo-formed minerals, excluding the influence of any

organic content in Azokh materials. However, further investigations using other techniques and a higher number of samples are needed.

7.The paleogenetic analysis of the coprolite yielded mitochondrial sequences identical to those of modern brown hyena (Hyaena brunnea) while the paleogenomic analysis did not reveal any indication for DNA sequences of a potential predator.

8.Brown hyenas are today and for their known history restricted to southern Africa, and it is unlikely that their range ever extended into Eurasia. The most parsimonious explanation for this result is contamination of the coprolite from fresh brown hyena scats that were treated in the University of the Free State laboratory in Bloemfontein prior to the opening of the coprolite.

9.In summary, none of the methods applied here has provided conclusive indication of the species that produced these coprolites. None of the results obtained could support or discard bears vs. hyenas as the taphonomic agent. Thus, the producer’s species cannot be defined at this time.

Acknowledgements We are grateful to the authorities of Nagorno-Karabakh for permissions to work on these specimens. We thank the Electron Microscopy Unit of the Museo Nacional de Ciencias Naturales for their careful and professional work, Teresa Sanz for pictures taken of the coprolites before processing and Pablo Silva for pictures of modern bears. The authors are also grateful to M.D. Pesquero for providing coprolite measurements from La Roma site. We thank Corinne Esser from the Zoo Fauverie du Mont Faron, France, for providing hair and scats of brown hyenas. The authors are grateful to comments from Mark Lewis, Nigel Larkin, the three anonymous reviewers and the editor in charge (Peter Andrews) who greatly improved this chapter.

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