iVikodak—A Platform and Standard Workflow for Inferring, Analyzing, Comparing, and Visualizing the Functional Potential of Microbial Communities

Background: The objectives of any metagenomic study typically include identification of resident microbes and their relative proportions (taxonomic analysis), profiling functional diversity (functional analysis), and comparing the identified microbes and functions with available metadata (comparative metagenomics). Given the advantage of cost-effectiveness and convenient data-size, amplicon-based sequencing has remained the technology of choice for exploring phylogenetic diversity of an environment. A recent school of thought, employing the existing genome annotation information for inferring functional capacity of an identified microbiome community, has given a promising alternative to Whole Genome Shotgun sequencing for functional analysis. Although a handful of tools are currently available for function inference, their scope, functionality and utility has essentially remained limited. Need for a comprehensive framework that expands upon the existing scope and enables a standardized workflow for function inference, analysis, and visualization, is therefore felt. Methods: We present iVikodak, a multi-modular web-platform that hosts a logically inter-connected repertoire of functional inference and analysis tools, coupled with a comprehensive visualization interface. iVikodak is equipped with microbial co-inhabitance pattern driven published algorithms along with multiple updated databases of various curated microbe-function maps. It also features an advanced task management and result sharing system through introduction of personalized and portable dashboards. Results: In addition to inferring functions from 16S rRNA gene data, iVikodak enables (a) an in-depth analysis of specific functions of interest (b) identification of microbes contributing to various functions (c) microbial interaction patterns through function-driven correlation networks, and (d) simultaneous functional comparison between multiple microbial communities. We have bench-marked iVikodak through multiple case studies and comparisons with existing state of art. We also introduce the concept of a public repository which provides a first of its kind community-driven framework for scientific data analytics, collaboration and sharing in this area of microbiome research. Conclusion: Developed using modern design and task management practices, iVikodak provides a multi-modular, yet inter-operable, one-stop framework, that intends to simplify the entire approach toward inferred function analysis. It is anticipated to serve as a significant value addition to the existing space of functional metagenomics. iVikodak web-server may be freely accessed at https://web.rniapps.net/iVikodak/.

[1]  Jesse R. Zaneveld,et al.  Predictive functional profiling of microbial communities using 16S rRNA marker gene sequences , 2013, Nature Biotechnology.

[2]  H. Bik,et al.  Coexisting cryptic species of the Litoditis marina complex (Nematoda) show differential resource use and have distinct microbiomes with high intraspecific variability , 2016, Molecular ecology.

[3]  M. Hattori,et al.  Multiple Omics Uncovers Host–Gut Microbial Mutualism During Prebiotic Fructooligosaccharide Supplementation , 2014, DNA research : an international journal for rapid publication of reports on genes and genomes.

[4]  Peter Williams,et al.  IMG: the integrated microbial genomes database and comparative analysis system , 2011, Nucleic Acids Res..

[5]  Gary D. Bader,et al.  Cytoscape.js: a graph theory library for visualisation and analysis , 2015, Bioinform..

[6]  Zhu-Hong You,et al.  A novel approach based on KATZ measure to predict associations of human microbiota with non‐infectious diseases , 2016, Bioinform..

[7]  Peter Meinicke,et al.  Tax4Fun: predicting functional profiles from metagenomic 16S rRNA data , 2015, Bioinform..

[8]  Ruth Ann Luna,et al.  Metagenomic pyrosequencing and microbial identification. , 2009, Clinical chemistry.

[9]  中尾 光輝,et al.  KEGG(Kyoto Encyclopedia of Genes and Genomes)〔和文〕 (特集 ゲノム医学の現在と未来--基礎と臨床) -- (データベース) , 2000 .

[10]  Karoline Faust,et al.  Soil microbiome responses to the short‐term effects of Amazonian deforestation , 2015, Molecular ecology.

[11]  Michael Y. Galperin,et al.  The COG database: a tool for genome-scale analysis of protein functions and evolution , 2000, Nucleic Acids Res..

[12]  Elizabeth M Glass,et al.  MG-RAST, a Metagenomics Service for Analysis of Microbial Community Structure and Function. , 2016, Methods in molecular biology.

[13]  Monzoorul Haque Mohammed,et al.  Microbial community profiling shows dysbiosis in the lesional skin of Vitiligo subjects , 2016, Scientific Reports.

[14]  Faber,et al.  Enzyme Classification and Nomenclature and Biocatalytic Retrosynthesis , 2015 .

[15]  Sharmila S. Mande,et al.  FLIM-MAP: Gene Context Based Identification of Functional Modules in Bacterial Metabolic Pathways , 2018, Front. Microbiol..

[16]  K. Rit,et al.  Prevalence of Pseudomonas aeruginosa and Acinetobacter spp producing metallo-β-lactamase in a tertiary care hospital , 2013 .

[17]  Martin J Blaser,et al.  Community differentiation of the cutaneous microbiota in psoriasis , 2013, Microbiome.

[18]  Jun Yin,et al.  Human Microbe-Disease Association Prediction Based on Adaptive Boosting , 2018, Front. Microbiol..

[19]  Luisa Delgado-Serrano,et al.  Respiratory tract clinical sample selection for microbiota analysis in patients with pulmonary tuberculosis , 2014, Microbiome.

[20]  James R. Cole,et al.  Ribosomal Database Project: data and tools for high throughput rRNA analysis , 2013, Nucleic Acids Res..

[21]  Yan Zhang,et al.  PATRIC, the bacterial bioinformatics database and analysis resource , 2013, Nucleic Acids Res..

[22]  Peer Bork,et al.  Enterotypes of the human gut microbiome , 2011, Nature.

[23]  William C. Gagne-Maynard,et al.  BURRITO: An interactive multi-omic tool for visualizing taxa-function relationships in microbiome data , 2017, bioRxiv.

[24]  Keith F. Tipton,et al.  Enzyme Classification and Nomenclature , 2001 .

[25]  C. Silva-Boghossian,et al.  Prevalence of Pseudomonas aeruginosa and Acinetobacter spp. in subgingival biofilm and saliva of subjects with chronic periodontal infection , 2014, Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology].

[26]  J Moran-Gilad,et al.  Practical issues in implementing whole-genome-sequencing in routine diagnostic microbiology. , 2017, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[27]  Zexuan Zhu,et al.  LRLSHMDA: Laplacian Regularized Least Squares for Human Microbe–Disease Association prediction , 2017, Scientific Reports.

[28]  Y. Benjamini,et al.  Controlling the false discovery rate: a practical and powerful approach to multiple testing , 1995 .

[29]  A. Heintz‐Buschart,et al.  IMP: a pipeline for reproducible integrated metagenomic and metatranscriptomic analyses , 2016, bioRxiv.

[30]  Jeffrey Heer,et al.  SpanningAspectRatioBank Easing FunctionS ArrayIn ColorIn Date Interpolator MatrixInterpola NumObjecPointI Rectang ISchedu Parallel Pause Scheduler Sequen Transition Transitioner Transiti Tween Co DelimGraphMLCon IData JSONCon DataField DataSc Dat DataSource Data DataUtil DirtySprite LineS RectSprite , 2011 .

[31]  William A. Walters,et al.  Using QIIME to Analyze 16S rRNA Gene Sequences from Microbial Communities , 2012, Current protocols in microbiology.

[32]  Eoin L. Brodie,et al.  Greengenes, a Chimera-Checked 16S rRNA Gene Database and Workbench Compatible with ARB , 2006, Applied and Environmental Microbiology.

[33]  Lutz Krause,et al.  Calypso: a user-friendly web-server for mining and visualizing microbiome–environment interactions , 2016, Bioinform..

[34]  J. A. Aas,et al.  Defining the Normal Bacterial Flora of the Oral Cavity , 2005, Journal of Clinical Microbiology.

[35]  Joon-Yong Lee,et al.  ATLAS (Automatic Tool for Local Assembly Structures) - a comprehensive infrastructure for assembly, annotation, and genomic binning of metagenomic and metatranscriptomic data , 2017, PeerJ Prepr..

[36]  Jeffrey Heer,et al.  D³ Data-Driven Documents , 2011, IEEE Transactions on Visualization and Computer Graphics.

[37]  Eric C. Li,et al.  Dysbiosis and Alterations in Predicted Functions of the Subgingival Microbiome in Chronic Periodontitis , 2014, Applied and Environmental Microbiology.

[38]  Owen White,et al.  The TIGRFAMs database of protein families , 2003, Nucleic Acids Res..

[39]  Yoav Freund,et al.  A decision-theoretic generalization of on-line learning and an application to boosting , 1995, EuroCOLT.

[40]  Tungadri Bose,et al.  COGNIZER: A Framework for Functional Annotation of Metagenomic Datasets , 2015, PloS one.

[41]  Susan Holmes,et al.  phyloseq: An R Package for Reproducible Interactive Analysis and Graphics of Microbiome Census Data , 2013, PloS one.

[42]  Bo Li,et al.  Antibiotic-induced shifts in the mouse gut microbiome and metabolome increase susceptibility to Clostridium difficile infection , 2014, Nature Communications.

[43]  Katherine H. Huang,et al.  Structure, Function and Diversity of the Healthy Human Microbiome , 2012, Nature.

[44]  David S. Wishart,et al.  METAGENassist: a comprehensive web server for comparative metagenomics , 2012, Nucleic Acids Res..

[45]  Shenjie Tang,et al.  Complex sputum microbial composition in patients with pulmonary tuberculosis , 2012, BMC Microbiology.

[46]  N. Segata,et al.  Shotgun metagenomics, from sampling to analysis , 2017, Nature Biotechnology.

[47]  S. Eddy,et al.  Pfam: the protein families database , 2013, Nucleic Acids Res..

[48]  M. Podar,et al.  Distinct and complex bacterial profiles in human periodontitis and health revealed by 16S pyrosequencing , 2011, The ISME Journal.

[49]  Xing Chen,et al.  Prediction of microbe–disease association from the integration of neighbor and graph with collaborative recommendation model , 2017, Journal of Translational Medicine.

[50]  Leo Breiman,et al.  Random Forests , 2001, Machine Learning.

[51]  Sharmila S. Mande,et al.  Vikodak - A Modular Framework for Inferring Functional Potential of Microbial Communities from 16S Metagenomic Datasets , 2016, PloS one.

[52]  P. Gajer,et al.  The vaginal microbiota of pregnant women who subsequently have spontaneous preterm labor and delivery and those with a normal delivery at term , 2014, Microbiome.

[53]  Liping Zhao,et al.  A gut microbiota-targeted dietary intervention for amelioration of chronic inflammation underlying metabolic syndrome , 2013, FEMS microbiology ecology.

[54]  W. Ludwig,et al.  SILVA: a comprehensive online resource for quality checked and aligned ribosomal RNA sequence data compatible with ARB , 2007, Nucleic acids research.