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      The elusive parasite: comparing macroscopic, immunological, and genomic approaches to identifying malaria in human skeletal remains from Sayala, Egypt (third to sixth centuries AD)

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          Abstract

          Although malaria is one of the oldest and most widely distributed diseases affecting humans, identifying and characterizing its presence in ancient human remains continue to challenge researchers. We attempted to establish a reliable approach to detecting malaria in human skeletons using multiple avenues of analysis: macroscopic observations, rapid diagnostic tests, and shotgun-capture sequencing techniques, to identify pathological changes, Plasmodium antigens, and Plasmodium DNA, respectively. Bone and tooth samples from ten individuals who displayed skeletal lesions associated with anaemia, from a site in southern Egypt (third to sixth centuries AD), were selected. Plasmodium antigens were detected in five of the ten bone samples, and traces of Plasmodium aDNA were detected in six of the twenty bone and tooth samples. There was relatively good synchronicity between the biomolecular findings, despite not being able to authenticate the results. This study highlights the complexity and limitations in the conclusive identification of the Plasmodium parasite in ancient human skeletons. Limitations regarding antigen and aDNA preservation and the importance of sample selection are at the forefront of the search for malaria in the past. We confirm that, currently, palaeopathological changes such as cribra orbitalia are not enough to be certain of the presence of malaria. While biomolecular methods are likely the best chance for conclusive identification, we were unable to obtain results which correspond to the current authentication criteria of biomolecules. This study represents an important contribution in the refinement of biomolecular techniques used; also, it raises new insight regarding the consistency of combining several approaches in the identification of malaria in past populations.

          Supplementary Information

          The online version contains supplementary material available at 10.1007/s12520-021-01350-z.

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          Most cited references119

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          The Sequence Alignment/Map format and SAMtools

          Summary: The Sequence Alignment/Map (SAM) format is a generic alignment format for storing read alignments against reference sequences, supporting short and long reads (up to 128 Mbp) produced by different sequencing platforms. It is flexible in style, compact in size, efficient in random access and is the format in which alignments from the 1000 Genomes Project are released. SAMtools implements various utilities for post-processing alignments in the SAM format, such as indexing, variant caller and alignment viewer, and thus provides universal tools for processing read alignments. Availability: http://samtools.sourceforge.net Contact: rd@sanger.ac.uk
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            Fast gapped-read alignment with Bowtie 2.

            As the rate of sequencing increases, greater throughput is demanded from read aligners. The full-text minute index is often used to make alignment very fast and memory-efficient, but the approach is ill-suited to finding longer, gapped alignments. Bowtie 2 combines the strengths of the full-text minute index with the flexibility and speed of hardware-accelerated dynamic programming algorithms to achieve a combination of high speed, sensitivity and accuracy.
              • Record: found
              • Abstract: found
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              Basic local alignment search tool.

              A new approach to rapid sequence comparison, basic local alignment search tool (BLAST), directly approximates alignments that optimize a measure of local similarity, the maximal segment pair (MSP) score. Recent mathematical results on the stochastic properties of MSP scores allow an analysis of the performance of this method as well as the statistical significance of alignments it generates. The basic algorithm is simple and robust; it can be implemented in a number of ways and applied in a variety of contexts including straightforward DNA and protein sequence database searches, motif searches, gene identification searches, and in the analysis of multiple regions of similarity in long DNA sequences. In addition to its flexibility and tractability to mathematical analysis, BLAST is an order of magnitude faster than existing sequence comparison tools of comparable sensitivity.

                Author and article information

                Contributors
                gamble.michelle@gmail.com
                Journal
                Archaeol Anthropol Sci
                Archaeol Anthropol Sci
                Archaeological and Anthropological Sciences
                Springer Berlin Heidelberg (Berlin/Heidelberg )
                1866-9557
                1866-9565
                14 June 2021
                14 June 2021
                2021
                : 13
                : 7
                : 115
                Affiliations
                [1 ]GRID grid.22937.3d, ISNI 0000 0000 9259 8492, Institute of Specific Prophylaxis and Tropical Medicine, Center for Pathophysiology, Infectiology and Immunology, , Medical University of Vienna, ; Kinderspitalgasse 15, 1090 Vienna, Austria
                [2 ]GRID grid.4299.6, ISNI 0000 0001 2169 3852, Bioarchaeology Department, , Austrian Archaeological Institute at the Austrian Academy of Sciences, ; Franz Klein-Gasse 1, 1190 Vienna, Austria
                [3 ]Present Address: Heritage and Archaeological Research Practice, 101 Rose Street South Lane, EH2 3JG Edinburgh, Scotland
                [4 ]GRID grid.418908.c, ISNI 0000 0001 1089 6435, Institute for Mummy Studies, , EURAC Research, ; Viale Druso 1, 39100 Bolzano, Italy
                [5 ]Present Address: Malaria Research Initiative Bandarban, Vienna, Austria
                [6 ]Present Address: Planen und Bauen im Bestand, Novetus, Belvederegasse 41, 1040 Vienna, Austria
                Author information
                http://orcid.org/0000-0003-4561-8026
                http://orcid.org/0000-0001-6282-4206
                http://orcid.org/0000-0001-5214-2615
                http://orcid.org/0000-0001-6946-9112
                http://orcid.org/0000-0003-2846-8994
                http://orcid.org/0000-0002-1461-747X
                http://orcid.org/0000-0002-3105-5878
                Article
                1350
                10.1007/s12520-021-01350-z
                8202054
                34149953
                d6d5ed0e-135c-4af3-a1df-efb2d5a4df40
                © The Author(s) 2021

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 16 December 2020
                : 19 April 2021
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100001822, Österreichischen Akademie der Wissenschaften;
                Award ID: IF_2015_12
                Award Recipient :
                Categories
                Original Paper
                Custom metadata
                © Springer-Verlag GmbH Germany, part of Springer Nature 2021

                malaria,immunoassays,shotgun-capture sequencing,adna,palaeopathology

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