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Methodological innovations for the study of irreplaceable samples reveal giardiasis in extinct animals (Nothrotherium maquinense and Palaeolama maior)
Authors:Daniela Leles  Paula Cascardo  Elisa Pucu  Beatriz Brener  Adriana Sudré  Elizabeth Alves  Flávia Uchoa  Priscilla Fajardo  Patrícia Millar  Danuza Mattos  Márcia Chame  Castor Cartelle
Institution:1. Laboratório de Biologia Molecular de Parasitos e de Paleoparasitologia, Departamento de Microbiologia e Parasitologia, Instituto Biomédico, Universidade Federal Fluminense, Rua Professor Hernani Melo, 101, São Domingos, Niterói-RJ CEP: 24210-130, Brazil;2. Escola Nacional de Saúde Pública Sérgio Arouca - Fundação Oswaldo Cruz, Rua Leopoldo Bulhões, 1480, Manguinhos, Rio de Janeiro-RJ CEP: 21041-210, Brazil;3. Universidade Estadual de Londrina, Rodovia Celso Garcia Cid PR445 Km 380, Londrina -PR. CEP: 86057-970, Brazil;4. Pontifícia Universidade Católica de Minas Gerais, Avenida Dom José Gaspar, 290 Coração Eucarístico, Belo Horizonte, Estado de Minas Gerais CEP: 30535-901, Brazil
Abstract:The use of diagnostic methods that prevent irreplaceable samples (from museum collections, archaeological and paleontological samples) of being consumed or that increase their yield is relevant. For museum collections, archaeological and paleontological samples it is essential to conserve samples, subsamples or portions for future research. We are addressing methods for conservation of irreplaceable samples that could be fully consumed. Innovations in methodologies that are used in studies of Paleoparasitology and Paleomicrobiology will contribute to the preservation of collections. Therefore, to the development of archaeology and paleontology in the future, we evaluated whether the discarded material of the immunochromatography test could be used for molecular diagnosis and vice versa. We used a genotyped experimental coprolite positive for Giardia duodenalis. The diagnosis was positive for giardiasis in both cases. This methodology can be corroborated with the coprolite of a Paleolama maior (extinct llama) previously diagnosed for G. duodenalis with an immunoenzymatic test. The residue of the pre-digestion step of the DNA extraction before adding Proteinase K was confirmed positive with the immunochromatographic test. Also, the DNA extraction residue from a coprolite of Nothrotherium maquinense (ground sloth) was tested positive with immunochromatographic test for G. duodenalis. These are the oldest findings for G. duodenalis confirming that this intestinal parasite occurred among Northeastern Brazilian Megafauna animals from the late Pleistocene period, correlated to human occupation. The relevance of these results will allow the study by different methodological approaches from a small amount of material, reusing discarded materials.
Keywords:Paleoparasitology  Coprolite  Immunodiagnosis  Ancient DNA  Megafauna  Corresponding author  
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