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phyloseq: An R Package for Reproducible Interactive Analysis and Graphics of Microbiome Census Data
Open accessAbstract
BACKGROUND: the analysis of microbial communities through dna sequencing brings many challenges: the integration of different types of data with methods from ecology, genetics, phylogenetics, multivariate statistics, visualization and testing. With the increased breadth of experimental designs now being pursued, project-specific statistical analyses are often needed, and these analyses are often difficult (or impossible) for peer researchers to independently reproduce. The vast majority of the requisite tools for performing these analyses reproducibly are already implemented in R and its extensions (packages), but with limited support for high throughput microbiome census data. RESULTS: Here we describe a software project, phyloseq, dedicated to the object-oriented representation and analysis of microbiome census data in R. It supports importing data from a variety of common formats, as well as many analysis techniques. These include calibration, filtering, subsetting, agglomeration, multi-table comparisons, diversity analysis, parallelized Fast UniFrac, ordination methods, and production of publication-quality graphics; all in a manner that is easy to document, share, and modify. We show how to apply functions from other R packages to phyloseq-represented data, illustrating the availability of a large number of open source analysis techniques. We discuss the use of phyloseq with tools for reproducible research, a practice common in other fields but still rare in the analysis of highly parallel microbiome census data. We have made available all of the materials necessary to completely reproduce the analysis and figures included in this article, an example of best practices for reproducible research. CONCLUSIONS: The phyloseq project for R is a new open-source software package, freely available on the web from both GitHub and Bioconductor.
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Unstable regulatory T cells, enriched for naïve and Nrp1negcells, are purged after fate challenge
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“Histogram tables and diversity indexes were obtained using the phyloseq package in Rstudio (McMurdie and Holmes, 2013).”
MethodUllern, A., Holm, K., Røssevold, A. H., Andresen, N. K., Bang, C., Lingjærde, O. C., Naume, B., Hov, J. R., & Kyte, J. A. (2025) · Molecular Oncology · doi:10.1002/1878-0261.13760 · Cited by 25
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“2 using the metagenomeSeq((20)), PhyloSeq((21)), Vegan: Community Ecology Package((22)) and GGplot2((23)) packages.”
InfluentialSequencing effort dictates gene discovery in marine microbial metagenomes
Duarte, C. M., Ngugi, D. K., Alam, I., Pearman, J., Kamau, A., Eguiluz, V. M., Gojobori, T., Acinas, S. G., Gasol, J. M., Bajic, V., et al. (2020) · Environmental Microbiology · doi:10.1111/1462-2920.15182 · Cited by 22
Socioeconomic position links circulatory microbiota differences with biological age
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Plastic biodegradation by sediment microbial populations under denitrifying conditions
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Universal Drivers of Cheese Microbiomes
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The Role of Intestinal Microbiota in Metastatic Colorectal Cancer Patients Treated With Capecitabine
Aarnoutse, R., Ziemons, J., de Vos-Geelen, J., Valkenburg-van Iersel, L., Wildeboer, A. C. L., Vievermans, A., Creemers, G.-J. M., Baars, A., Vestjens, H. J. H. M. J., Le, G. N., et al. (2022) · Clinical Colorectal Cancer · doi:10.1016/j.clcc.2021.10.004 · Cited by 19
Associations between Escherichia coli O157 shedding and the faecal microbiota of dairy cows
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Le Luyer, J., Schull, Q., Auffret, P., Lopez, P., Crusot, M., Belliard, C., Basset, C., Carradec, Q., Poulain, J., Planes, S., et al. (2021) · Animal Microbiome · doi:10.1186/s42523-021-00097-1 · Cited by 16
Shimba, A., Cui, G., Abe, S., Hirota, K., Miyauchi, E., Takami, D., Tani-ichi, S., Kato, R., Tajima, M., Kanahashi, T., et al. (2025) · Cell Reports · doi:10.1016/j.celrep.2025.116093 · Cited by 15
Investigation into host selection of the cecal acetogen population in rabbits after weaning.
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Valles-Colomer, M., Manghi, P., Cumbo, F., Masetti, G., Armanini, F., Asnicar, F., Blanco-Miguez, A., Pinto, F., Punčochář, M., Garaventa, A., et al. (2024) · eBioMedicine · doi:10.1016/j.ebiom.2023.104917 · Cited by 14
“31 Species-level richness (Observed) and diversity (Simpson ’ s) were determined using the estimate_richness function in phyloseq on the untransformed species-level abundance MetaPhlAn matrix, and evenness (Pielou) was calculated as the Shannon ’ s diversity divided by the logarithm of the Observed…”
MethodKharel, M., Tian, M., Vu, T., Miao, Y., & Zhang, W. (2025) · Journal of Hazardous Materials · doi:10.1016/j.jhazmat.2025.139754 · Cited by 13
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Investigating microbial and environmental drivers of nitrification in alkaline forest soil
Poghosyan, L., & Lehtovirta-Morley, L. E. (2024) · ISME Communications · doi:10.1093/ismeco/ycae093 · Cited by 10
Fluctuating selection on bacterial iron regulation in the mammalian gut
Barreto, H. C., Abreu, B., & Gordo, I. (2022) · Current Biology · doi:10.1016/j.cub.2022.06.017 · Cited by 10
Host Species and Environment Shape the Skin Microbiota of Mexican Axolotls
Soto-Cortés, E., Marroquín-Rodríguez, M., Basanta, M. D., Maldonado-López, Y., Parra-Olea, G., & Rebollar, E. A. (2024) · Microbial Ecology · doi:10.1007/s00248-024-02411-1 · Cited by 9
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Assessing the Influence of Dietary History on Gut Microbiota
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Comprehensive Metagenomic Analysis of Veterinary Probiotics in Broiler Chickens
Kerek, Á., Román, I. L., Szabó, Á., Papp, M., Bányai, K., Kardos, G., Kaszab, E., Bali, K., Makrai, L., & Jerzsele, Á. (2024) · Animals · doi:10.3390/ani14131927 · Cited by 8
Buttimer, S., Medina, D., Martins, R. A., da Silva, A. G. M., Neely, W. J., Haddad, C. F. B., DiRenzo, G. V., Catenazzi, A., Bell, R. C., & Becker, C. G. (2025) · Global Change Biology · doi:10.1111/gcb.70275 · Cited by 8
Anitua, E., Murias-Freijo, A., Tierno, R., Tejero, R., & Alkhraisat, M. H. (2024) · BMC Oral Health · doi:10.1186/s12903-024-04675-y · Cited by 7
Integrative analysis of taste genetics and the dental plaque microbiome in early childhood caries
Khan, M. W., Cruz de Jesus, V., Mittermuller, B.-A., Schroth, R. J., Hu, P., & Chelikani, P. (2025) · Cell Reports · doi:10.1016/j.celrep.2025.116245 · Cited by 7
Microbiome depiction through user-adapted bioinformatic pipelines and parameters
Nayman, E. I., Schwartz, B. A., Polanco, F. C., Firek, A. K., Gumabong, A. C., Hofstee, N. J., Narasimhan, G., Cickovski, T., & Mathee, K. (2023) · Journal of Medical Microbiology · doi:10.1099/jmm.0.001756 · Cited by 6
Yeo, K., Wu, F., Li, R., Smith, E., Wormald, P.-J., Valentine, R., Psaltis, A. J., Vreugde, S., & Fenix, K. (2024) · Pathogens · doi:10.3390/pathogens13100826 · Cited by 5
Stage-Specific Microbiota Transitions Throughout Black Soldier Fly Ontogeny
Klammsteiner, T., Heussler, C. D., Stonig, K. T., Insam, H., Schlick-Steiner, B. C., & Steiner, F. M. (2026) · Microbial Ecology · doi:10.1007/s00248-025-02691-1 · Cited by 3
Parente, E., Zotta, T., & Ricciardi, A. (2022) · doi:10.1101/2022.01.19.476946 · Cited by 15
Effects of silver diamine fluoride on oral bacteriome and mycobiome: a randomized clinical trial
Manerkar, M., de Jesus, V. C., Mittermuller, B.-A., Lee, V. H. K., Singh, S., Bertone, M., Chelikani, P., & Schroth, R. J. (2025) · BMC Oral Health · doi:10.1186/s12903-025-06986-0 · Cited by 2
Microbiome as a Tool to Monitor Aquarium Systems
Elmagzoub, W. A., Weidmann, M., Elnaiem, M. H. E., Dennig, A., Waller, U., Bernhard, A., Junhold, J., Abd El Wahed, A., Truyen, U., & Ceruti, A. (2026) · Veterinary Sciences · doi:10.3390/vetsci13020125 · Cited by 1
Metadata
- Authors
- McMurdie, P. J., & Holmes, S.
- Venue
- PLoS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2013-04-22
- Volume
- 8
- Issue
- 4
- Pages
- e61217
- ISSN
- 1932-6203 (ISSN-L)
- OpenAlex
- W2056279562
- Cited by
- 21,322
- References
- 11
- FWCI
- 58.63
- Type
- journal article
- Language
- en
- Open access
- gold
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Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.