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Studies on drug metabolism by fungi colonizing decomposing human cadavers. Part I: DNA sequence-based identification of fungi isolated from postmortem material

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Abstract

Cadavers can be colonized by a wide variety of bacteria and fungi. Some of these microbes could change the concentration or the metabolic pattern of drugs present in postmortem samples. The purpose of this study was to identify fungi from human postmortem material and to further assess their potential role in the metabolism of drugs. Aliquots of 252 postmortem samples (heart blood, liver, kidney, and lung) taken from 105 moderately to severely decomposed bodies were streaked on Sabouraud agar for isolation of fungal species. One part of the samples was worked up immediately after autopsy (group I). The second part had previously been stored at −20 °C for at least 1 year (group II). Identification of the isolates was achieved morphologically by microscopy and molecularly by polymerase chain reaction amplification and sequencing of markers allowing species identification of the respective genera. Depending on the genus, different gene fragments were used: calmodulin for Aspergillus, β-tubulin for Penicillium, translation elongation factor 1α for Fusarium, and the internal transcribed spacer region of the ribosomal DNA for all remaining genera. A total of 156 fungal strains were isolated from 62 % of the postmortem materials. By using these primers, 98 % of the isolates could be identified to the species level. The most common genera were Candida (60.0 %—six species), Penicillium (10.3 %—two species), Rhodotorula (7.1 %—one species), Mucor (6.4 %—four species), Aspergillus (3.2 %—four species), Trichosporon (3.2 %—one species), and Geotrichum (3.2 %—one species). Group I samples contained 53 % more fungal species than stored samples suggesting some fungi did not survive the freezing process. The isolated fungi might be characteristic for decomposed bodies. The proposed methodology proved to be appropriate for the identification of fungi in this type of material.

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Acknowledgments

Jorge Martinez thanks DAAD for the support of this project and the Gesellschaft für Toxikologische und Forensische Chemie (Society of Toxicological and Forensic Chemistry, GTFCh) for providing the travel fund to present part of this work at the 50th Annual Meeting of the International Association of Forensic Toxicologists, June 3–8, 2012, Hamamatsu, Japan. The authors also thank Ricarda Arnold, Stephanie Drobnik, Sabine Eska, Reinhard Heiderstädt, Juliane Höfig, Cornelia Jacob, Adelheid Mattern, Sabine Müller, Christian Ortmann, Friederike Raab, Julia Schermer, and Ulrich Schmatloch for their assistance with sampling and DNA sequencing as well as Daniela Remane for proofreading of the manuscript.

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Authors and Affiliations

  1. Institute of Forensic Medicine, Jena University Hospital, Fürstengraben 23, 07743, Jena, Germany

    Jorge A. Martínez-Ramírez, Juliane Strien, Juliane Sanft, Gita Mall & Frank T. Peters

  2. Department of Pharmacy, National University, A.A. 14490, Bogotá D.C., Colombia

    Jorge A. Martínez-Ramírez

  3. Institute of Microbiology, Department of Microbiology and Molecular Biology, University of Jena, Neugasse 25, 07743, Jena, Germany

    Grit Walther

  4. Leibniz-Institute for Natural Product Research and Infection Biology—Hans-Knöll-Institute, Jena Microbial Resource Collection, Beutenbergstr. 11a, 07745, Jena, Germany

    Grit Walther

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  1. Jorge A. Martínez-Ramírez
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Correspondence to Frank T. Peters.

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Martínez-Ramírez, J.A., Strien, J., Sanft, J. et al. Studies on drug metabolism by fungi colonizing decomposing human cadavers. Part I: DNA sequence-based identification of fungi isolated from postmortem material. Anal Bioanal Chem 405, 8443–8450 (2013). https://doi.org/10.1007/s00216-013-7250-1

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