scholarly journals Reference-based error correction of amplicon sequencing data from synthetic communities

2021 ◽  
Author(s):  
Pengfan Zhang ◽  
Stjin Spaepen ◽  
Yang Bai ◽  
Stephane Hacquard ◽  
Ruben Garrido-Oter
Keyword(s):  
Microbial Communities ◽  
Amplicon Sequencing ◽  
R Package ◽  
Fungal Communities ◽  
Polymorphic Variation ◽  
Sequencing Data ◽  
Extensive Evaluation ◽  
Culture Independent ◽  
Reference Sequences ◽  
Synthetic Microbial Communities

AbstractMotivationSynthetic microbial communities (SynComs) constitute an emergent and powerful tool in biological, biomedical, and biotechnological research. Despite recent advances in algorithms for analysis of culture-independent amplicon sequencing data from microbial communities, there is a lack of tools specifically designed for analysing SynCom data, where reference sequences for each strain are available.ResultsHere we present Rbec, a tool designed for analysing SynCom data that outperforms current methods by accurately correcting errors in amplicon sequences and identifying intra-strain polymorphic variation. Extensive evaluation using mock bacterial and fungal communities show that our tool performs robustly for samples of varying complexity, diversity, and sequencing depth. Further, Rbec also allows accurate detection of contaminations in SynCom experiments.AvailabilityRbec is freely available as an open-source R package and can be downloaded at: https://github.com/PengfanZhang/Microbiome.

scholarly journals Rbec: a tool for analysis of amplicon sequencing data from synthetic microbial communities

2021 ◽  
Vol 1 (1) ◽  
Author(s):  
Pengfan Zhang ◽  
Stjin Spaepen ◽  
Yang Bai ◽  
Stephane Hacquard ◽  
Ruben Garrido-Oter
Keyword(s):  
Microbial Communities ◽  
Amplicon Sequencing ◽  
Fungal Communities ◽  
Polymorphic Variation ◽  
Sequencing Data ◽  
Sequencing Errors ◽  
Extensive Evaluation ◽  
Culture Independent ◽  
Reference Sequences ◽  
Synthetic Microbial Communities

AbstractSynthetic microbial communities (SynComs) constitute an emerging and powerful tool in biological, biomedical, and biotechnological research. Despite recent advances in algorithms for the analysis of culture-independent amplicon sequencing data from microbial communities, there is a lack of tools specifically designed for analyzing SynCom data, where reference sequences for each strain are available. Here we present Rbec, a tool designed for the analysis of SynCom data that accurately corrects PCR and sequencing errors in amplicon sequences and identifies intra-strain polymorphic variation. Extensive evaluation using mock bacterial and fungal communities show that our tool outperforms current methods for samples of varying complexity, diversity, and sequencing depth. Furthermore, Rbec also allows accurate detection of contaminants in SynCom experiments.

Comparative fungal community analyses using metatranscriptomics and ITS-amplicon sequencing from Norway spruce

2021 ◽  
Author(s):  
Andreas Schneider ◽  
John Sundh ◽  
Görel Sundström ◽  
Kerstin Richau ◽  
Nicolas Delhomme ◽  
...  
Keyword(s):  
Community Structure ◽  
Microbial Communities ◽  
Norway Spruce ◽  
Environmental Samples ◽  
Marker Gene ◽  
Amplicon Sequencing ◽  
Fungal Communities ◽  
Individual Species ◽  
Rna Seq ◽  
Sequencing Data

<p>Microbial communities are major players in carbon and nitrogen cycling globally and are of particular importance for plant communities in the nutrient poor soils of boreal forests. Especially relevant are the fungal communities in the soil that interact with the plants in multiple ways, indirectly through their pivotal role in the breakdown of organic matter and, more directly, through mycorrhizal symbiosis with plant roots. Large-scale disturbances of these complex microbial communities can lead to shifts in soil carbon storage with unknown and global-scale long-term consequences. To understand the dynamics of these communities and their relationship to associated plants in response to climate change and anthropogenic influence, we need a better understanding of how modern “omics” methods can help us to understand compositional and functional shifts of these microbiomes. Microbial gene expression and functional activity can be assayed with RNA sequencing (RNA-Seq) data from environmental samples. In contrast, currently phylogenetic marker gene amplicon sequencing data is generally used to assess taxonomic composition and community structure of the microbiome. Few studies have considered how much of this structural and taxonomic information is included in RNA-Seq transcriptomic data from matched samples. Here we describe fungal communities using both RNA-Seq and fungal ITS1 DNA amplicon sequencing to compare the outcomes between the methods. We used a panel of root and needle samples from mature stands of the coniferous tree species Picea abies (Norway spruce) growing in untreated (nutrient deficient) and nutrient enriched plots at the Flakaliden forest research site in boreal northern Sweden. We created an assembly-based, reproducible and hardware agnostic workflow to taxonomically and functionally annotate fungal RNA-Seq data obtained from Norway spruce roots, which we compared to matching ITS amplicon sequencing data.<strong> </strong>We show that the community structure indicated by the fungal transcriptome is in agreement with that generated by the ITS data, while also identifying limitations imposed by current database coverage. Furthermore, we show examples to demonstrate how metatranscriptomics data additionally provides biologically informative functional insight at the community and individual species level. These findings highlight the potential of metatranscriptomics to advance our understanding of interaction, response and effect both between host plants and their associated microbial communities, and among the members of microbial communities in environmental samples in general.</p>

scholarly journals The response of soil and phyllosphere microbial communities to repeated application of the fungicide iprodione: accelerated biodegradation or toxicity?

2020 ◽  
Vol 96 (6) ◽  
Author(s):  
A Katsoula ◽  
S Vasileiadis ◽  
M Sapountzi ◽  
Dimitrios G Karpouzas
Keyword(s):  
Microbial Communities ◽  
Agricultural Production ◽  
Soil Microorganisms ◽  
Amplicon Sequencing ◽  
Fungal Communities ◽  
Similar Response ◽  
Repeated Application ◽  
Plant Leaves ◽  
Soil System ◽  
Plant Soil

ABSTRACT Pesticides interact with microorganisms in various ways with the outcome being negative or positive for the soil microbiota. Pesticides' effects on soil microorganisms have been studied extensively in soil but not in other pesticides-exposed microbial habitats like the phyllosphere. We tested the hypothesis that soil and phyllosphere support distinct microbial communities, but exhibit a similar response (accelerated biodegradation or toxicity) to repeated exposure to the fungicide iprodione. Pepper plants received four repeated foliage or soil applications of iprodione, which accelerated its degradation in soil (DT50_1st = 1.23 and DT50_4th = 0.48 days) and on plant leaves (DT50_1st > 365 and DT50_4th = 5.95 days). The composition of the epiphytic and soil bacterial and fungal communities, determined by amplicon sequencing, was significantly altered by iprodione. The archaeal epiphytic and soil communities responded differently; the former showed no response to iprodione. Three iprodione-degrading Paenarthrobacter strains were isolated from soil and phyllosphere. They hydrolyzed iprodione to 3,5-dichloraniline via the formation of 3,5-dichlorophenyl-carboxiamide and 3,5-dichlorophenylurea-acetate, a pathway shared by other soil-derived arthrobacters implying a phylogenetic specialization in iprodione biotransformation. Our results suggest that iprodione-repeated application could affect soil and epiphytic microbial communities with implications for the homeostasis of the plant–soil system and agricultural production.

scholarly journals A high-resolution pipeline for 16S-sequencing identifies bacterial strains in human microbiome

2019 ◽  
Author(s):  
Igor Segota ◽  
Tao Long
Keyword(s):  
Bacterial Species ◽  
Human Microbiome ◽  
Amplicon Sequencing ◽  
R Package ◽  
Strain Level ◽  
Sequencing Data ◽  
Bacterial Strains ◽  
16S Sequencing ◽  
16S Amplicon Sequencing ◽  
Sequencing Data Analysis

We developed a High-resolution Microbial Analysis Pipeline (HiMAP) for 16S amplicon sequencing data analysis, aiming at bacterial species or strain-level identification from human microbiome to enable experimental validation for causal effects of the associated bacterial strains on health and diseases. HiMAP achieved higher accuracy in identifying species in human microbiome mock community than other pipelines. HiMAP identified majority of the species, with strain-level resolution wherever possible, as detected by whole genome shotgun sequencing using MetaPhlAn2 and reported comparable relative abundances. HiMAP is an open-source R package available at https://github.com/taolonglab/himap.

scholarly journals Microbial Communities on Plastic Polymers in the Mediterranean Sea

2021 ◽  
Vol 12 ◽  
Author(s):  
Annika Vaksmaa ◽  
Katrin Knittel ◽  
Alejandro Abdala Asbun ◽  
Maaike Goudriaan ◽  
Andreas Ellrott ◽  
...  
Keyword(s):  
Microbial Communities ◽  
Mediterranean Sea ◽  
Amplicon Sequencing ◽  
Polymer Surfaces ◽  
Rrna Gene ◽  
Sequencing Data ◽  
Microbial Biofilms ◽  
Degrading Bacteria ◽  
The Mediterranean

Plastic particles in the ocean are typically covered with microbial biofilms, but it remains unclear whether distinct microbial communities colonize different polymer types. In this study, we analyzed microbial communities forming biofilms on floating microplastics in a bay of the island of Elba in the Mediterranean Sea. Raman spectroscopy revealed that the plastic particles mainly comprised polyethylene (PE), polypropylene (PP), and polystyrene (PS) of which polyethylene and polypropylene particles were typically brittle and featured cracks. Fluorescence in situ hybridization and imaging by high-resolution microscopy revealed dense microbial biofilms on the polymer surfaces. Amplicon sequencing of the 16S rRNA gene showed that the bacterial communities on all plastic types consisted mainly of the orders Flavobacteriales, Rhodobacterales, Cytophagales, Rickettsiales, Alteromonadales, Chitinophagales, and Oceanospirillales. We found significant differences in the biofilm community composition on PE compared with PP and PS (on OTU and order level), which shows that different microbial communities colonize specific polymer types. Furthermore, the sequencing data also revealed a higher relative abundance of archaeal sequences on PS in comparison with PE or PP. We furthermore found a high occurrence, up to 17% of all sequences, of different hydrocarbon-degrading bacteria on all investigated plastic types. However, their functioning in the plastic-associated biofilm and potential role in plastic degradation needs further assessment.

scholarly journals In ovo microbial communities: a potential mechanism for the initial acquisition of gut microbiota among oviparous birds and lizards

2018 ◽  
Vol 14 (7) ◽  
pp. 20180225 ◽  
Author(s):  
Brian K. Trevelline ◽  
Kirsty J. MacLeod ◽  
Sarah A. Knutie ◽  
Tracy Langkilde ◽  
Kevin D. Kohl
Keyword(s):  
Gut Microbiota ◽  
Microbial Communities ◽  
Bacterial Communities ◽  
Reproductive Tract ◽  
Egg Yolk ◽  
Fungal Communities ◽  
In Ovo ◽  
Physiological Processes ◽  
Initial Acquisition ◽  
Culture Independent

Vertebrate gut microbiota mediate critical physiological processes known to affect host fitness, but the mechanisms that expose wildlife to pioneer members of this important microbial community are not well understood. For example, oviparous vertebrates are thought to acquire gut microbiota through post-natal exposure to the external environment, but recent evidence from placental mammals suggests that the vertebrate reproductive tract harbours microbiota that may inoculate offspring in utero . These findings suggest that oviparous vertebrates may be capable of acquiring pioneer microbiota in ovo , but this phenomenon remains unexplored. To fill this knowledge gap, we used culture-independent inventories to determine if the eggs of wild birds and lizards harboured in ovo microbial communities. Our approach revealed distinct in ovo bacterial communities, but fungal communities were indistinguishable from controls. Further, lizard eggs from the same clutch had bacterial community structures that were more similar to each other than to unrelated individuals. These results suggest that oviparous vertebrates may acquire maternal microbiota in ovo , possibly through the inoculation of egg yolk prior to shelling. Therefore, this study may provide a first glimpse of a phenomenon with substantial implications for our understanding of the ecological and evolutionary factors shaping gut microbial communities.

scholarly journals animalcules: interactive microbiome analytics and visualization in R

Microbiome ◽  
2021 ◽  
Vol 9 (1) ◽  
Author(s):  
Yue Zhao ◽  
Anthony Federico ◽  
Tyler Faits ◽  
Solaiappan Manimaran ◽  
Daniel Segrè ◽  
...  
Keyword(s):  
16S Rrna ◽  
Microbial Communities ◽  
R Package ◽  
Command Line ◽  
Data Generation ◽  
Sequencing Data ◽  
Shotgun Metagenomics ◽  
Microbiome Analysis ◽  
Link Type ◽  
R Shiny

Abstract Background Microbial communities that live in and on the human body play a vital role in health and disease. Recent advances in sequencing technologies have enabled the study of microbial communities at unprecedented resolution. However, these advances in data generation have presented novel challenges to researchers attempting to analyze and visualize these data. Results To address some of these challenges, we have developed animalcules, an easy-to-use interactive microbiome analysis toolkit for 16S rRNA sequencing data, shotgun DNA metagenomics data, and RNA-based metatranscriptomics profiling data. This toolkit combines novel and existing analytics, visualization methods, and machine learning models. For example, the toolkit features traditional microbiome analyses such as alpha/beta diversity and differential abundance analysis, combined with new methods for biomarker identification are. In addition, animalcules provides interactive and dynamic figures that enable users to understand their data and discover new insights. animalcules can be used as a standalone command-line R package or users can explore their data with the accompanying interactive R Shiny interface. Conclusions We present animalcules, an R package for interactive microbiome analysis through either an interactive interface facilitated by R Shiny or various command-line functions. It is the first microbiome analysis toolkit that supports the analysis of all 16S rRNA, DNA-based shotgun metagenomics, and RNA-sequencing based metatranscriptomics datasets. animalcules can be freely downloaded from GitHub at https://github.com/compbiomed/animalcules or installed through Bioconductor at https://www.bioconductor.org/packages/release/bioc/html/animalcules.html.

microeco: An R package for data mining in microbial community ecology

2020 ◽  
Author(s):  
Chi Liu ◽  
Yaoming Cui ◽  
Xiangzhen Li ◽  
Minjie Yao
Keyword(s):  
Data Mining ◽  
Microbial Community ◽  
Community Ecology ◽  
Amplicon Sequencing ◽  
R Package ◽  
Environmental Data ◽  
Venn Diagram ◽  
Diversity Analysis ◽  
Sequencing Data ◽  
Microbial Community Ecology

Abstract A large amount of sequencing data is produced in microbial community ecology studies using the high-throughput sequencing technique, especially amplicon-sequencing-based community data. After conducting the initial bioinformatic analysis of amplicon sequencing data, performing the subsequent statistics and data mining based on the operational taxonomic unit and taxonomic assignment tables is still complicated and time-consuming. To address this problem, we present an integrated R package-‘microeco’ as an analysis pipeline for treating microbial community and environmental data. This package was developed based on the R6 class system and combines a series of commonly used and advanced approaches in microbial community ecology research. The package includes classes for data preprocessing, taxa abundance plotting, venn diagram, alpha diversity analysis, beta diversity analysis, differential abundance test and indicator taxon analysis, environmental data analysis, null model analysis, network analysis and functional analysis. Each class is designed to provide a set of approaches that can be easily accessible to users. Compared with other R packages in the microbial ecology field, the microeco package is fast, flexible and modularized to use, and provides powerful and convenient tools for researchers. The microeco package can be installed from CRAN (The Comprehensive R Archive Network) or github (https://github.com/ChiLiubio/microeco).

scholarly journals Animalcules: Interactive Microbiome Analytics and Visualization in R

2020 ◽  
Author(s):  
Yue Zhao ◽  
Anthony Federico ◽  
Tyler Faits ◽  
Solaiappan Manimaran ◽  
Stefano Monti ◽  
...  
Keyword(s):  
16S Rrna ◽  
Microbial Communities ◽  
R Package ◽  
Vital Role ◽  
Command Line ◽  
Data Generation ◽  
Sequencing Data ◽  
Shotgun Metagenomics ◽  
Microbiome Analysis ◽  
R Shiny

Abstract Background: Microbial communities that live in and on the human body play a vital role in health and disease. Recent advances in sequencing technologies have enabled the study of microbial communities at unprecedented resolution. However, these advances in data generation have presented novel challenges to researchers attempting to analyze and visualize these data.Results: To address some of these challenges, we have developed Animalcules, an easy-to-use interactive microbiome analysis toolkit for 16S rRNA sequencing data, shotgun DNA metagenomics data, and RNA-based metatranscriptomics profiling data. This toolkit combines novel and existing analytics, visualization methods, and machine learning models. For example, traditional microbiome analyses such as alpha/beta diversity and differential abundance analysis are enhanced in the toolkit, while new methods such as biomarker identification are introduced. Powerful interactive and dynamic figures generated by Animalcules enable users to understand their data and discover new insights. Animalcules can be used as a standalone command-line R package or users can explore their data with the accompanying interactive R Shiny interface.Conclusions: We present Animalcules, an R package for interactive microbiome analysis through either an interactive interface facilitated by R Shiny or various command-line functions. It is the first microbiome analysis toolkit that supports the analysis of all 16S rRNA, DNA-based shotgun metagenomics, and RNA-sequencing based metatranscriptomics datasets. Animalcules can be freely downloaded from GitHub at https://github.com/compbiomed/animalcules or installed through Bioconductor (https://www.bioconductor.org/packages/release/bioc/html/animalcules.html).

MicroNiche: an R package for assessing microbial niche breadth and overlap from amplicon sequencing data

2020 ◽  
Vol 96 (8) ◽  
Author(s):  
D R Finn ◽  
J Yu ◽  
Z E Ilhan ◽  
V M C Fernandes ◽  
C R Penton ◽  
...  
Keyword(s):  
In Silico ◽  
Niche Breadth ◽  
Biological Soil Crust ◽  
A Priori ◽  
Amplicon Sequencing ◽  
R Package ◽  
Training Data ◽  
Sequencing Data ◽  
Human Gut ◽  
Limit Of Quantification

ABSTRACT Niche is a fundamental concept in ecology. It integrates the sum of biotic and abiotic environmental requirements that determines a taxon's distribution. Microbiologists currently lack quantitative approaches to address niche-related hypotheses. We tested four approaches for the quantification of niche breadth and overlap of taxa in amplicon sequencing datasets, with the goal of determining generalists, specialists and environmental-dependent distributions of community members. We applied these indices to in silico training datasets first, and then to real human gut and desert biological soil crust (biocrust) case studies, assessing the agreement of the indices with previous findings. Implementation of each approach successfully identified a priori conditions within in silico training data, and we found that by including a limit of quantification based on species rank, one could identify taxa falsely classified as specialists because of their low, sparse counts. Analysis of the human gut study offered quantitative support for Bacilli, Gammaproteobacteria and Fusobacteria specialists enriched after bariatric surgery. We could quantitatively characterise differential niche distributions of cyanobacterial taxa with respect to precipitation gradients in biocrusts. We conclude that these approaches, made publicly available as an R package (MicroNiche), represent useful tools to assess microbial environment-taxon and taxon-taxon relationships in a quantitative manner.

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