Algorithms for scalable and efficient population genomics and metagenomics

Microbes strongly impact human health and the ecosystem of which they are a part. Rapid improvements and decreasing costs in sequencing technologies have revolutionized the field of genomics and enabled important insights into microbial genome biology and microbiomes. However, new tools and approaches are needed to facilitate the efficient analysis of large sets of genomes and to associate genomic features with phenotypic characteristics better. Here, we built and utilized several tools for large-scale whole-genome analysis for different microbial characteristics, such as antimicrobial resistance and pathogenicity, that are important for human health. Chapters 2 and 3 demonstrate the needs and challenges of population genomics in associating antimicrobial resistance with genomic features. Our results highlight important limitations of reference database-driven analysis for genotype-phenotype association studies and demonstrate the utility of whole-genome population genomics in uncovering novel genomic factors associated with antimicrobial resistance. Chapter 4 describes PRAWNS, a fast and scalable bioinformatics tool that generates compact pan-genomic features. Existing approaches are unable to meet the needs of large-scale whole-genome analyses, either due to scalability limitations or the inability of the genomic features generated to support a thorough whole-genome assessment. We demonstrate that PRAWNS scales to thousands of genomes and provides a concise collection of genomic features which support the downstream analyses. In Chapter 5, we assess whether the combination of long and short-read sequencing can expedite the accurate reconstruction of a pathogen genome from a microbial community. We describe the challenges for pathogen detection in current foodborne illness outbreak monitoring. Our results show that the recovery of a pathogen genome can be accelerated using a combination of long and short-read sequencing after limited culturing of the microbial community. We evaluated several popular genome assembly approaches and identified areas for improvement. In Chapter 6, we describe SIMILE, a fast and scalable bioinformatics tool that enables the detection of genomic regions shared between several assembled metagenomes. In metagenomics, microbial communities are sequenced directly without culturing. Although metagenomics has furthered our understanding of the microbiome, comparing metagenomic samples is extremely difficult. We describe the need and challenges in comparing several metagenomic samples and present an approach that facilitates large-scale metagenomic comparisons

NCBI’s virus discovery codeathon: building “FIVE” —the Federated Index of Viral Experiments API index

Viruses represent important test cases for data federation due to their genome size and the rapid increase in sequence data in publicly available databases. However, some consequences of previously decentralized (unfederated) data are lack of consensus or comparisons between feature annotations. Unifying or displaying alternative annotations should be a priority both for communities with robust entry representation and for nascent communities with burgeoning data sources. To this end, during this three-day continuation of the Virus Hunting Toolkit codeathon series (VHT-2), a new integrated and federated viral index was elaborated. This Federated Index of Viral Experiments (FIVE) integrates pre-existing and novel functional and taxonomy annotations and virus–host pairings. Variability in the context of viral genomic diversity is often overlooked in virus databases. As a proof-of-concept, FIVE was the first attempt to include viral genome variation for HIV, the most well-studied human pathogen, through viral genome diversity graphs. As per the publication of this manuscript, FIVE is the first implementation of a virus-specific federated index of such scope. FIVE is coded in BigQuery for optimal access of large quantities of data and is publicly accessible. Many projects of database or index federation fail to provide easier alternatives to access or query information. To this end, a Python API query system was developed to enhance the accessibility of FIVE

Current progress and future opportunities in applications of bioinformatics for biodefense and pathogen detection: report from the Winter Mid-Atlantic Microbiome Meet-up, College Park, MD, January 10, 2018

Abstract The Mid-Atlantic Microbiome Meet-up (M3) organization brings together academic, government, and industry groups to share ideas and develop best practices for microbiome research. In January of 2018, M3 held its fourth meeting, which focused on recent advances in biodefense, specifically those relating to infectious disease, and the use of metagenomic methods for pathogen detection. Presentations highlighted the utility of next-generation sequencing technologies for identifying and tracking microbial community members across space and time. However, they also stressed the current limitations of genomic approaches for biodefense, including insufficient sensitivity to detect low-abundance pathogens and the inability to quantify viable organisms. Participants discussed ways in which the community can improve software usability and shared new computational tools for metagenomic processing, assembly, annotation, and visualization. Looking to the future, they identified the need for better bioinformatics toolkits for longitudinal analyses, improved sample processing approaches for characterizing viruses and fungi, and more consistent maintenance of database resources. Finally, they addressed the necessity of improving data standards to incentivize data sharing. Here, we summarize the presentations and discussions from the meeting, identifying the areas where microbiome analyses have improved our ability to detect and manage biological threats and infectious disease, as well as gaps of knowledge in the field that require future funding and focus