Biology subjects

McHardy, A. C.

Publications and source records attributed to McHardy, A. C..

8 recordsLinked to original sources

bioRxiv2018

Assessing taxonomic metagenome profilers with OPAL

Taxonomic metagenome profilers predict the presence and relative abundance of microorganisms from shotgun sequence samples of DNA isolated directly from a microbial community. Over the past years, there has been an explosive growth of software and algorithms for this task, resulting in a need for more systematic comparisons of these methods based on relevant performance criteria. Here, we present OPAL, a software package implementing commonly used performance metrics, including those of the first challenge of the Initiative for the Critical Assessment of Metagenome Interpretation (CAMI), together with convenient visualizations. In addition, OPAL implements diversity metrics from microbial ecology, as well as run time and memory efficiency measurements. By allowing users to customize the relative importance of metrics, OPAL facilitates in-depth performance comparisons, as well as the development of new methods and data analysis workflows. To demonstrate the application, we compared seven profilers on benchmark datasets of the first and second CAMI challenges using all metrics and performance measurements available in OPAL. The software is implemented in Python 3 and available under the Apache 2.0 license on GitHub (https://github.com/CAMI-challenge/OPAL).\n\nAuthor summaryThere are many computational approaches for inferring the presence and relative abundance of taxa (i.e. taxonomic profiling) from shotgun metagenome samples of microbial communities, making systematic performance evaluations a very important task. However, there has yet to be introduced a computational framework in which profiler performances can be compared. This delays method development and applied studies, as researchers need to implement their own custom evaluation frameworks. Here, we present OPAL, a software package that facilitates standardized comparisons of taxonomic metagenome profilers. It implements a variety of performance metrics frequently employed in microbiome research, including runtime and memory usage, and generates comparison reports and visualizations. OPAL thus facilitates and accelerates benchmarking of taxonomic profiling techniques on ground truth data. This enables researchers to arrive at informed decisions about which computational techniques to use for specific datasets and research questions.

bioinformatics↗

bioRxiv2018

Structures and functions linked to genome-wide adaptation of human influenza A viruses

Human influenza A viruses elicit short-term respiratory infections with considerable mortality and morbidity. While H3N2 viruses circulate since almost 50 years, the recent introduction of pH1N1 viruses presents an excellent opportunity for a comparative analysis of the genome-wide evolutionary forces acting on both subtypes. Here, we inferred patches of sites relevant for adaptation, i.e. being under positive selection, on eleven viral protein structures, from all available data since 1968 and correlated these with known functional properties. Overall, pH1N1 had more patches than H3N2 viruses, especially in the viral polymerase complex, while antigenic evolution is more apparent for H3N2 viruses. In both subtypes, NS1 had the highest patch and patch site frequency, indicating that NS1-mediated viral attenuation of host inflammatory responses is a continuously intensifying process, elevated even in the longtime-circulating subtype H3N2. We confirmed the resistance-causing effects of two pH1N1 changes against oseltamivir in NA activity assays, demonstrating the value of the resource for discovering functionally relevant changes. Our results represent an atlas of protein regions and sites with links to host adaptation, antiviral drug resistances and immune evasion for both subtypes for further study.

bioinformatics↗

bioRxiv2018

Nucleotide-pair encoding of 16S rRNA sequences for host phenotype and biomarker detection

Identifying combinations of taxa distinctive for microbiome-associated diseases is considered key to the establishment of diagnosis and therapy options in precision medicine and imposes high demands on accuracy of microbiome analysis techniques. We propose subsequence based 16S rRNA data analysis, as a new paradigm for microbiome phenotype classification and biomarker detection. This method and software called DiTaxa substitutes standard OTU-clustering or sequence-level analysis by segmenting 16S rRNA reads into the most frequent variable-length subsequences. These subsequences are then used as data representation for downstream phenotype prediction, biomarker detection and taxonomic analysis. Our proposed sequence segmentation called nucleotide-pair encoding (NPE) is an unsupervised data-driven segmentation inspired by Byte-pair encoding, a data compression algorithm. The identified subsequences represent commonly occurring sequence portions, which we found to be distinctive for taxa at varying evolutionary distances and highly informative for predicting host phenotypes. We compared the performance of DiTaxa to the state-of-the-art methods in disease phenotype prediction and biomarker detection, using human-associated 16S rRNA samples for periodontal disease, rheumatoid arthritis and inflammatory bowel diseases, as well as a synthetic benchmark dataset. DiTaxa identified 17 out of 29 taxa with confirmed links to periodontitis (recall= 0.59), relative to 3 out of 29 taxa (recall= 0.10) by the state-of-the-art method. On synthetic benchmark data, DiTaxa obtained full precision and recall in biomarker detection, compared to 0.91 and 0.90, respectively. In addition, machine-learning classifiers trained to predict host disease phenotypes based on the NPE representation performed competitively to the state-of-the art using OTUs or k-mers. For the rheumatoid arthritis dataset, DiTaxa substantially outperformed OTU features with a macro-F1 score of 0.76 compared to 0.65. Due to the alignment- and reference free nature, DiTaxa can efficiently run on large datasets. The full analysis of a large 16S rRNA dataset of 1359 samples required {approx}1.5 hours on 20 cores, while the standard pipeline needed {approx}6.5 hours in the same setting.\n\nAvailabilityAn implementation of our method called DiTaxa is available under the Apache 2 licence at http://llp.berkeley.edu/ditaxa.

bioinformatics↗

bioRxiv2018

CAMISIM: Simulating metagenomes and microbial communities

Shotgun metagenome data sets of microbial communities are highly diverse, not only due to the natural variation of the underlying biological systems, but also due to differences in laboratory protocols, replicate numbers, and sequencing technologies. Accordingly, to effectively assess the performance of metagenomic analysis software, a wide range of benchmark data sets are required. Here, we describe the CAMISIM microbial community and metagenome simulator. The software can model different microbial abundance profiles, multi-sample time series and differential abundance studies, includes real and simulated strain-level diversity, and generates second and third generation sequencing data from taxonomic profiles or de novo. Gold standards are created for sequence assembly, genome binning, taxonomic binning, and taxonomic profiling. CAMSIM generated the benchmark data sets of the first CAMI challenge. For two simulated multi-sample data sets of the human and mouse gut microbiomes we observed high functional congruence to the real data. As further applications, we investigated the effect of varying evolutionary genome divergence, sequencing depth, and read error profiles on two popular metagenome assemblers, MEGAHIT and metaSPAdes, on several thousand small data sets generated with CAMISIM. CAMISIM can simulate a wide variety of microbial communities and metagenome data sets together with truth standards for method evaluation. All data sets and the software are freely available at: https://github.com/CAMI-challenge/CAMISIM

bioinformatics↗

bioRxiv2018

MicroPheno: Predicting environments and host phenotypes from 16S rRNA gene sequencing using a k-mer based representation of shallow sub-samples

MotivationMicrobial communities play important roles in the function and maintenance of various biosystems, ranging from the human body to the environment. A major challenge in microbiome research is the classification of microbial communities of different environments or host phenotypes. The most common and cost-effective approach for such studies to date is 16S rRNA gene sequencing. Recent falls in sequencing costs have increased the demand for simple, efficient, and accurate methods for rapid detection or diagnosis with proved applications in medicine, agriculture, and forensic science. We describe a reference- and alignment-free approach for predicting environments and host phenotypes from 16S rRNA gene sequencing based on k-mer representations that benefits from a bootstrapping framework for investigating the sufficiency of shallow sub-samples. Deep learning methods as well as classical approaches were explored for predicting environments and host phenotypes.\n\nResultsk-mer distribution of shallow sub-samples outperformed the computationally costly Operational Taxonomic Unit (OTU) features in the tasks of body-site identification and Crohn's disease prediction. Aside from being more accurate, using k-mer features in shallow sub-samples allows (i) skipping computationally costly sequence alignments required in OTU-picking, and (ii) provided a proof of concept for the sufficiency of shallow and short-length 16S rRNA sequencing for phenotype prediction. In addition, k-mer features predicted representative 16S rRNA gene sequences of 18 ecological environments, and 5 organismal environments with high macro-F1 scores of 0.88 and 0.87. For large datasets, deep learning outperformed classical methods such as Random Forest and SVM.\n\nAvailabilityThe software and datasets are available at https://llp.berkeley.edu/micropheno.

bioinformatics↗

bioRxiv2017

AMBER: Assessment of Metagenome BinnERs

Reconstructing the genomes of microbial community members is key to the interpretation of shotgun metagenome samples. Genome binning programs deconvolute reads or assembled contigs of such samples into individual bins, but assessing their quality is difficult due to the lack of evaluation software and standardized metrics. We present AMBER, an evaluation package for the comparative assessment of genome reconstructions from metagenome benchmark data sets. It calculates the performance metrics and comparative visualizations used in the first benchmarking challenge of the Initiative for the Critical Assessment of Metagenome Interpretation (CAMI). As an application, we show the outputs of AMBER for ten different binnings on two CAMI benchmark data sets. AMBER is implemented in Python and available under the Apache 2.0 license on GitHub (https://github.com/CAMI-challenge/AMBER).

bioinformatics↗

bioRxiv2017

Fast and memory-efficient noisy read overlapping with KD-trees

MotivationThird-generation sequencing technologies produce long, but noisy reads with increasing sequencing throughput and decreasing per-base costs. Detecting read-to-read overlaps in such data is the most computationally intensive step in de novo assembly. Recently, efficient algorithms were developed for this task; nearly all of these utilize long k-mers (>10 bp) to compare reads, but vary in their approaches to indexing, hashing, filtering, and dimensionality reduction.\n\nResultsWe describe an algorithm for efficient overlap detection that directly compares the full spectrum of short k-mers, namely tetramers, through geometric embedding and approximate nearest neighbor search in multidimensional KD-trees. A proof of concept implementation detected read-to-read overlaps in bacterial PacBio and ONT datasets with notably lower memory consumption than state-of-the-art approaches and allowed downstream de novo assembly into single contigs. We also introduce a sequence-context dependent tagging scheme that contributes to memory and computational efficiency and could be used with other aligning and overlapping algorithms.\n\nAvailabilityA C++14 implementation is available under the open source Apache License 2.0 at: https://github.com/dzif/kd-tree-overlapper

bioinformatics↗

bioRxiv2017

Laboratory cultivation of acidophilic nanoorganisms. Physiological and bioinformatic dissection of a stable laboratory co-culture.

This study describes the laboratory cultivation of ARMAN (Archaeal Richmond Mine Acidophilic Nanoorganisms). After 2.5 years of successive transfers in an anoxic medium containing ferric sulfate as an electron acceptor, a consortium was attained that is comprised of two members of the order Thermoplasmatales, a member of a proposed ARMAN group, as well as a fungus. The 16S rRNA of one archaeon is only 91.6% identical to Thermogymnomonas acidicola as most closely related isolate. Hence, this organism is the first member of a new genus. The enrichment culture is dominated by this microorganism and the ARMAN. The third archaeon in the community seems to be present in minor quantities and has a 100% 16S rRNA identity to the recently isolated Cuniculiplasma divulgatum. The enriched ARMAN species is most probably incapable of sugar metabolism because the key genes for sugar catabolism and anabolism could not be identified in the metagenome. Metatranscriptomic analysis suggests that the TCA cycle funneled with amino acids is the main metabolic pathway used by the archaea of the community. Microscopic analysis revealed that growth of the ARMAN is supported by the formation of cell aggregates. These might enable cross feeding by other community members to the ARMAN.

microbiology↗