Lahun Pyramid, More Surprising Treasures Are Revealed: Fungal Diversity Analysis Using Amplicon-BasedMetagenomic Approach

Author:

S. Soliman, M. Magdy

Author address:

Ain Shams University, CAIRO, Egypt

Full conference title:

20th Congress of the International Society for Human and Animal Mycology, Amsterdam, the Netherlands

Date: 24 July 2018

Abstract:

RIF are commonly known as cultural-heritage bio-deteriorating agents. The mud-brick Lahun pyramid of Egypt located on the west bank of the Nile valley, is believed to be constructed by the pharaoh Senusret II (12th dynasty) 4,000 years ago. Recently, vividly painted wooden coffins housing an anthology of pharaonic-era mummies were uncovered, the mummies are the first to be discovered beneath the sand surrounding the pyramid in the depths of the desert rock. As a part of long-term survey for RIF inhabiting Egyptian monuments to record its occurrences and distribution range, ITS-based metagenomic analysis of samples from Lahun Pyramid site was performed. Methods: Samples were collected from the pyramid remains, DNA (eDNA) was extracted using PowerSoil DNA Isolation kit (MO BIO), successful extractions were quantified, and quality checked prior to be sequenced using MiSeq3000 (Macrogen, Inc., Korea). ITSx was used to filter ITS2 sequences to improve classification precision. Filtered reads were classified using RDP and Taxonomer tools for metagenomic analysis based on UNITE dataset. Species were counted and filtered based on similarity index (>90%) not on relative abundance, because melanized fungi generally yield low DNA quantities due to its cell walls nature. Results: Total of 17k filtered reads were successfully classified as fungi (61%) in 118 species representing four phyla, Ascomycota (63%), Basidiomycota (35%), Glomeromycota (1%) and Zygomycota (2%). The highest species number per group belongs to class Sordariomycetes (27). Among many, two black yeasts described as RIF were detected namely are: Auriopasidium nambiae, Coniosporium apollinus. Moreover, eight black yeast species described as human pathogens were detected: Capronia coronate, Cladophialophora immunda, Cyphellophora europea, Exophiala xenobiotica Exophiala oligosperma and Fonsecaea multimorphosa (family Herpotrichiellaceae) and Blastomyces dermatitidis and Histoplasma capsulatum (family Ajellomycetaceae). Histoplasma capsulatum that is most often associated with river valleys and Myceliophthora thermophila that need high-temperature to grow (45-50 ’) fit within the site conditions. However, unexpected classified species never been reported to the site conditions was detected, for example, Pseudozyma antarctica that was initially isolated from Antarctica as Candida antarctica. Conclusion: Contrasting modern genomics against traditional methods, using NGS based sequencing approach is very useful, especially for some cases. For example, Wallemia ichthyophaga never grows below 1.5MNaCl supplemented media, which usually is not the case for the general fungal isolation media. The approach sensitivity is only challenged by the error rates and sequences artifacts, which is being handled using bioinformatic tools and advanced pipelines to overcome this technical problem. On the other hand, the success of eDNA extraction from hyper-arid regions is not common, while reporting the success of ITS-based metagenomic technique application to it, is worth reporting. The current study reflected higher fungal diversity than expected, successfully detected a bio-deteriorating agent to a cultural heritage site and efficiently provided a prior information for traditional isolation methods to isolate and validate the detected species for further studies.

Abstract Number: S8.2d

Conference Year: 2018

Link to conference website:

Link Conference abstract: 

ISHAM 2018

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