HISTOLOGICAL AND PALEOGENETIC ANALYSES OF TWO PRE-COLUMBIAN MUMMIES

Transcription

1 HISTOLOGICAL AND PALEOGENETIC ANALYSES OF TWO PRE-COLUMBIAN MUMMIES GAETA Raffaele 1, FORNACIARI Gino 1, GIUFFRA Valentina 1, MAROTA Isolina 2, SANTIAGO- RODRIGUEZ Tasha 3, CANO Raul 4 1 Division of Paleopathology, Department of Translational Research and of New Technologies in in Medicine and Surgery, University of Pisa,Italy 2 Laboratory of Molecular Archaeo-Anthropology/ancient DNA, School of Biosciences and Veterinary Medicine, University of Camerino, Italy 3 Institute for Life Science Entrepreneurship, ATCC-Center for Translational Microbiology, Union, NJ, United States 4 The BioCollective, Denver, CO, United States 30 th European Congress of Pathology ESP Bilbao September 8-12, 2018

2 The Museum of Anthropology and Ethnology of the University of Florence houses the collection of some South-American mummies, carried in Italy between 1868 and 1914 by anthropologists like Enrico Hillyer Giglioli, Oscar Perrone, Beniamino Calcagno and Ernesto Mazzei. Palazzo Nonfinito, site of the Museum Display case with South American mummies

3 FI12 Natural mummy in discrete state of preservation of a young woman (20±5 years) lying in fetal position. Probably deposited in a basket made of woven vegetable ropes. 14 C analysis dated the body in a range between 1410 and 1530.

4 TC examination showed no pathological alterations of the skeleton. The internal organs were poorly preserved. A complete autopsy was then performed.

5 The right lung shows numerous alveoli, in large fibrous areas, with well recognizable vessels and bronchi Histological results Widespread areas of pulmonary anthracosis (H&E, 40X) An important element is the massive presence of exudate. (H&E, 100X)

6 (H&E, 40X) In the left lung there are residues of pulmonary fibrous texture; the alveolar pattern is still clearly visible. We can observe the tissue with lacunae containing a dense material. visible even in Masson's trichrome stain. (MAS, 25X)

7 The woman suffered from EXUDATiVE PNEUMONIA characterized by: - congestion of parieto-alveolar capillaries - Intra-alveolar serous exudate (eosinophilic material rich in proteins) Ancient VS Modern

8 FI9 Natural mummy of a young woman (20±3 years) from Cuzco, the ancient capital of the Inca empire. The case of woven vegetable ropes is typical of the Late Intermediate Period ( A.D.) and 14 C dating is between AD.

9 Macroscopical examination revealed a megavisceral syndrome characterized by: Macroscopical results Gastric ectasia Megacolon with enormous amount of feces and Megacolon Megacolon with feces (arrows)

10 megaesophagus marked cardiomegaly Cardiomegaly Cardiomegaly (yellow arrow = megaesophagus)

11 (H&E, 50X) (H&E, 100X) Cardiac vessels with parasite embolism (right)

12 Immunohistochemistry Immunohistochemistry was performed with monoclonar antibodies anti-t. cruzi a-flagellar FCH-F8-1 and FCH- F8-4, product by the Instituto Fatala Chaben in Buenos Aires. The oval formations showed an intense immunoreactivity. (Esophagus, 50X) These aspects were compatible with microcysts of Trypanosoma cruzi amastigotes (Esophagus, 400X)

13 (Giemsa, 1000X) (Giemsa, 1000X) Intramyocardial amastigotes of Trypanosoma cruzi Intramyocardial amastigotes (modern specimen)

14 1. Bacterial populations Paleogenetic results: Gut Microbioma Gut microbiome The natural mummification process favours the preservation of Firmicutes (thick cell walls, spores, endospores) and the genus Clostridium is among the most abundant... Bacterial taxonomy based on 16rRNA gene at the genus level for paleofeces (Panel A), total colon (Panel B), descending colon (Panel C), transverse colon (Panel D), ascending colon...but the high representation of Clostridium may also be a reflection of a pathological state (were pre-columbian people exposed to pathogens by ingestion of contaminated food or water?)

15 2. Human Papilloma virus (HPV) Phylogenetic reconstruction of ancient and modern HPV types 21 and 49 E1 and/or E2 The phylogeny of HPV types 21 and 49 shows that HPV-49 is very similar to modern HPV-49, instead HPV-21 is different, probably by different adaptation of these two strains to the Spanish colonization.

16 FI3 Mummy of an adult male from Cuzco, Perù (14-15 century).

17 3. Trypanosoma cruzi Phylogenetic reconstruction of the alpha ribosomal subunit of ancient and modern T. cruzi CL Brener Compared with the sequences of modern T. cruzi, the FI12 pathogen appears more ancient than that of the other two mummies. The T. Cruzi is ancient and was an important and widespread cause of morbidity and mortality in pre-columbian civilization. A previous study (Aufderheide et al, 2004) found sequences of tripanosoma in 40.6% of South American mummies.

18 4. Leishmania donovani Phylogenetic reconstruction of ancient and modern Leishmania Donovani Ld39 Compared with the sequences of modern Leishmania D., the FI3 pathogen appears more ancient than that of FI9. However both mummies have a similarity of 97% with the modern type of Leishmania. Leishmania D. has a remote origin in the Americas and is previous to the European colonization.

19 5. Antibiotic resistance Percentage of putative antibiotic-resistance genes in mummified gut. Panel A. Percentage of genes associated with beta-lactamases, penicillin-binding proteins, resistance to fosfomycin, chloramphenicol, aminoglycosides, sulfa, quinolones, tetracycline and vancomycin. Panel B. Percentage of vancomycin genes including vana, B, H, T, Y R and S

20 5. Antibiotic resistance Antibiotic-resistance is ancient and originates from the environment (Da Costa, 2011, Nature) Resistance genes, pre-dating the modern antibiotic selective therapies, could have been caused by the presence of natural antibiotics

21 Pre-Columbian mummies Aragonese mummies (Naples, XVI century) VS

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23 The human gut is a reservoir of Antibiotic-Resistance Genes (ARG). High proportions of ARG in pre-inca mummies and modern Amazonians compared to Italian Renaissance nobility suggest that a lifestyle with a great exposure to the environment and consumption of soil-contaminated crops may result in a higher acquisition of antibiotic resistance

24 Raffaele Gaeta, MD, PhD student Unit of Anatomic Pathology 1 Division of Paleopathology Department of Translational Research, University of Pisa Laboratories at Medical School (Pisa) Via Paradisa Pisa - Italy Tel raffaele.gaeta@med.unipi.it