Symposium 5: Host-parasite relationships and diseases: lessons from the past (Aug 10, 2016)


9th WORLD CONGRESS ON MUMMY STUDIES
10-13 AUGUST 2016  
LIMA, PERU     

Wednesday 10 August, 2016

Symposium 5: Host-parasite relationships and diseases: lessons from the past

Chairs: Raffaella Bianucci, Karl Reinhard, Dong Hoon Shin
Time: 2:00 pm - 6:00 pm 
Ejecutivo II Room, 1st floor

(1) 2:20 Karl Reinhard, Isabel Teixeira-Santos, Elisa Pucu: Archaeological parasitology based on mummy diagnosis: a new approach. (Read by Elisa Pucu) 

(2) 2:40 Elisa Pucu, Paula Cascardo, Marcia Chame, Gisele Daltrini Felice, Niède Guidon, Maria Cleonice Vergne, Daniela Leles : Sensibility and specificity of primers designed long before new sequences were deposited: a critical approach applied to the archaeological context.

(3) 3:00 Alena  Iñiguez, Mônica Vieira, Lucélia Guedes, Morgana Camacho, Victor Hugo Borba, Alexandre Fernandes, Sergio Miranda, Karl Reinhard, Adauto Araújo: Paleoparasitology in Brazil: status and perspectives.

3:20 Discussion

3:30 COFFEE BREAK

(4) 4:00 Chang S. Oh, Min Seo, Ho Chul Ki, Jong-Yil Chai, Dong Hoon Shin. Current trends of paleoparasitology in Korean mummy studies.

(5) 4:20 Alena  Iñiguez, Herminia Gijón-Botella, María del Carmen del Arco-Aguilar, Mercedes Martín-Oval, Conrado Rodríguez-Martín, Mercedes del Arco-Aguilar, Adauto Araújo† Guanche mummies: integrating paleoparasitological and paleogenetic investigations.

(6) 4:40 Shênia P. C. Novo, Daniela Leles,  Adauto Araújo†, Raffaella Bianucci: Identification of Leishmania tarentolae signature in a post-Colonial Brazilian human mummy reopens questions on its ability to survive and spread systematically in human hosts.

(7) 5:00 Kelly Harkins, Lars Fehren-Schmitz: Challenges of using NGS to detect T. cruzi in human remains from pre-Columbian South America.

5:20 Discussion

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ABSTRACTS


(1) Archaeological Parasitology: A New Approach 

Karl Reinhard, University of Nebraska - Lincoln 
Isabel Teixeira-Santos, University of Nebraska - Lincoln
Elisa Pucu de Araujo, Universidade Federal Fluminense, Niterói

The discovery of a mummy with the gross pathology of megacolon in the Lower Pecos Canyonlands showed that Trypanosoma cruzi transmission was present 1,000 years ago.  By itself, this discovery was interesting, but isolated case.  However, in context of subsistence strategy for the region, it provided a basis for generating hypotheses that could be tested by archaeological research.  In 2014, Araújo and Reinhard published a research strategy for field recovery of evidence of T. cruzi transmission, both sylvatic and domiciled.  They propose a three-pronged approach of sampling archaeological middens for triatomines, sampling woodrat middens, and testing human coprolites for T. cruzi DNA.  They suggested that bulk samples, especially from rockshelters, be recovered from stratigraphic deposits to examine for insects. It is probable that rockshelter deposits will preserve intact or partially fragmented insects.  They also suggested that woodrat bones be carefully recovered and retained for molecular analysis.  Woodrats are the common definitive host for T. cruzi and were a major part of the hunter-gatherer diet.  Finally, they suggested that human corporeal remains, and coprolites, be tested for T. cruzi via ELISA and molecular means.  Our analysis of the first season of excavations began in January of 2016.   The excavations focused on Eagle Nest Cave, near Langtry, Texas. This presentation reports the results of preliminary investigations.


(2) Sensibility and specificity of primers designed long before new sequences were deposited: a critical approach applied to the archaeological context

Elisa Pucu1, Paula Cascardo1, Marcia Chame2, Gisele Daltrini Felice3,4, Niede Guidon3 and Daniela Leles1
1- Departamento de Microbiologia e Parasitologia, Instituto Biomédico, Universidade Federal Fluminense (UFF), Niterói, RJ, Brasil
2- Escola Nacional de Saúde Pública, Fiocruz, Rio de Janeiro, RJ, Brasil
3- Fundação Museu do Homem Americano, São Raimundo Nonato, PI, Brasil 
4- Universidade Federal do Vale do São Francisco (UNIVASF), Campus Serra de Capivara, São Raimundo Nonato–PI, Brasil

The recovery of ancient DNA opens the possibility of studying human-parasite relationships. Not only we still need to deal with the general issues of preservation, contamination and decay of samples, but whether the parasite DNA is still in the archaeological sample. Some primers specificity might be put to a test due to the universe of new information and the daily sequencing of organisms. In 1996, Souto and colleagues defined two lineages of Trypanosoma cruzi amplified of part of the intergenic region of the mini exon genes. These primers were deposited before other organisms had their DNA sequenced. In 2008, two papers were published using these primers in human remains with successful results. We will describe the results of using T. cruzi primers with archaeological samples and discuss the possibility of updating primers regularly as more organisms are being sequenced. Twelve bones from Sergipe collected by Cleonice Vergne were analyzed: one amplified Propionibacterium acnes, and one amplified Agrobacterium tumefaciens. P.acnes is a bacterium found in the human skin known to maintain the inflammatory phase of acne. Its genome sequence was published in 2004. A.tumefaciens occurs in the soil and it is a plant pathogen. Also, five bone samples of the extinct ground sloth from Piauí were tested. We found similarity with Pseudomonas putida, a non-pathogenic bacteria that can degrade synthetic materials. Few studies reported cases when primers amplify different organisms. However, even though the results are different from expected they should not be discarded as they might reveal unexpected results endogenous to the archaeological sample.



(3) PALEOPARASITOLOGY IN BRAZIL: STATUS AND PERSPECTIVES

Alena Mayo Iñiguez, Mônica Viera, Lucelia Guedes, Morgana Camacho, Vitor Hugo Borba, Alexandre Fernandes, Sergio Miranda, Karl Reinhard, Adauto Araujo

Paleoparasitology, the study of parasites found in archaeological materials, is a term originated in Brazil by Dr. Luiz Fernando Ferreira, at Oswaldo Cruz Foundation-Fiocruz. As a branch of Parasitology, and of Paleopathology, was created with the main objective of studding the origin and evolution of parasitic infections, based on a broad concept of parasites including viruses, bacteria, protozoa, helminths, and other forms of life with ecological niche in a host. Paleoparasitological findings have changed the previously accepted conceptions about the origin of infectious diseases in the New World. Our research group has demonstrated that the presence of geohelminths in pre-Columbian populations could be considered an evidence of alternative routes of the peopling of America. And also that American Trypanosomiases and Chagas disease, was present before the arrival of Europeans, for example.
The group is establishing methods and techniques to study human and animal parasites from both archaeological and paleontological remains. Especially in material from the littoral as Brazilian sambaquis (shell mounds), where parasites are rare. Paleoparasitological analysis have been conducting in archaeological sites from Brazil and worldwide. New records of parasites from pre-Columbian archaeological sites from Chile and USA will be presented. The understanding of the introduction/spread of new parasites and/or new lineages with the arrival of European and African slaves in America is an important topic in our research. In this regard, we have evaluated the presence of intestinal parasites in human remains from archaeological sites of the colonial period in Brazil. Historical sites, with different cultural and epidemiological contexts, showing similar parasite diversity in spite of divergence in prevalence. We have working on integrating paleoparasitological, and paleogenetic analysis, not only for parasites but also for hosts, and now with palynology and geoprocessing data, for a more comprehensive paleoepidemiological scenery.

(4) Current Trends of Paleoparasitology in Korean Mummy Studies 

Chang Seok Oh*, Min Seo**, Ho Chul Ki*, Jong-Yil Chai***, and Dong Hoon Shin* 

*Bioanthropology and Paleopathology Lab; ***Department of Parasitology and Tropical Medicine, Seoul National University College of Medicine, South Korea 
**Department of Parasitology, Dankook University, Chonan, South Korea 

Paleoparasitology reveals the prevalence of each parasite infection of the past, by examination on the samples from archaeological sites. Especially as for coprolites obtained from Korean mummy, as its preservation status was quite good for parasitological analysis, invaluable information about the parasitism in history could be accumulated. Briefly, microscopic examination confirmed the presence of parasite eggs in Korean mummy’s coprolites, by which we estimate the infection prevalence of specific parasitism among pre-modern Korean people. Using ancient DNA technique, we can reconstruct each parasite’s phylogenetic trees from a historical perspective. The sociocultural background of the parasitism in pre-modern Korean society was also studied lately. By intense collaboration with historians who know much about what was going on the health and disease status in pre-modern society, we tried to make a hypothesis about the presumptive route of ancient parasite infection in Korea, especially on Ascaris, Trichuris, Clonorchis, Metagonimus, Paragonimus, and Taenia spp. By such interdisciplinary collaboration between different research fields, we can get invaluable academic clues for comprehending ancient parasitism in pre-modern Korean history. 

(5) GUANCHE MUMMIES: INTEGRATING PALEOPARASITOLOGICAL AND PALEOGENETIC INVESTIGATIONS

Alena Mayo Iñiguez, Herminia Gijón-Botella, María del Carmen del Arco-Aguilar, Mercedes Martín-Oval, Conrado Rodríguez-Maffiotte, Mercedes del Arco-Aguilar§, Adauto Araújo 


The Guanches, ancient inhabitants of the Canary Islands, Spain, practiced mummification of their dead. A paleoparasitological and paleogenetic analysis was conducted on mummified bodies (n = 6) (AD 1200, Cal BP 750) belonging to the Guanche culture from Gran Canaria Island (Figure 1). Coprolite and sediment samples (n = 19) were removed from below the abdominal region or sacral foramina (Table 1). The samples were rehydrated in 0.5% trisodium phosphate solution for 72 hr at 4°C, and the paleoparasitological investigation was conducted by spontaneous sedimentation method and microscopic examination. The results revealed the presence of well-preserved eggs of Ascaris sp., Trichuris trichiura, Enterobius vermicularis, and hookworms (Figure 2; Table 2). Ancient DNA was extracted from sediment samples to elucidate the ancestry of the mummies and for molecular detection of Ascaris sp. infection. Results of paleogenetic analysis demonstrated Ascaris sp. infection using 2 molecular targets, cytb and nad1. mtDNA haplotypes U6b, U6b1, and HV were identified that confirmed records of Guanche ancestry (Table 3). The excellent preservation of Guanche mummies facilitated the paleoparasitological and paleogenetic study, the results of which contribute to our knowledge of Guanche culture and their health status.

(6) Identification of Leishmania tarentolae signature in a post-colonial Brazilian human mummy re-opens questions on its ability to survive and spread systematiclly in human hosts.

Novo S.P.1, Leles D2,  Aráujo A.3†, Bianucci R. 4,5

1 Departamento de Ciências Biológicas, Escola Nacional de Saúde Pública Sergio Arouca, Fundação Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brasil. E-mail: shenia@ensp.fiocruz.br
2Departamento de Microbiologia e Parasitologia, Instituto Biomédico, Universidade Federal Fluminense, Laboratório de Biologia Molecular de Parasitos, Niterói, Rio de Janeiro, Brazil. E-mail: dleles@id.uff.br
3† Departamento de Endemias Samuel Pessoa, Escola Nacional de Saúde Pública Sergio Arouca, Fundação Oswaldo Cruz, Fiocruz, Rio de Janeiro, Brasil. 
4Department of Public Health and Paediatric Sciences, Legal Medicine Section, University of Turin, Turin, Italy. E-mail: raffaella.bianucci@unito.it 
5UMR 7268, Laboratoire d’Anthropologie bio-culturelle, Droit, Etique & Santé (Adés), Faculté de Médecine de Marseille, France.

Leishmaniases are a complex of diseases with a broad clinical spectrum and epidemiological diversity. Caused by different species of the genus Leishmania, they represent a major threat to public health in different areas of the world.  Advances in paleoparasitology allowed scholars to recover remnants of ancient Leishmania parasites and to identify biological signatures in various hosts. Here we report on the identification of L. tarentolae signatures in soft and hard tissues of a Brazilian male dating to the post-colonial period (18th-19th c A.D.) from Minas Gerais. 
L. tarentolae is a Sauroleishmania that originates from the Old World and is currently not pathogenic to humans. Used in vaccines, L. tarentolae is apparently unable to complete its life cycle within the human host. Our findings contradict the previous statement and re-open the discussion concerning the survival potential of a L. tarentolae ancient strains in human macrophages, their ability to spread systematically and cause a disease. These data, in turn, raise a series of questions concerning the parasite’s way of transmission from lizards to humans, its presence in the Brazilian paleoenvironment and in the ancient lizard population.


Key words: Lizard populations, Minas Gerais, Old World, Brazilian paleoenvironment

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