scholarly journals StrainPro – a highly accurate Metagenomic strain-level profiling tool

2019 ◽  
Author(s):  
Hsin-Nan Lin ◽  
Yaw-Ling Lin ◽  
Wen-Lian Hsu

ABSTRACTCharacterizing the taxonomic diversity of a microbial community is very important to understand the roles of microorganisms. Next generation sequencing (NGS) provides great potential for investigation of a microbial community and leads to Metagenomic studies. NGS generates DNA fragment sequences directly from microorganism samples, and it requires analysis tools to identify microbial species (or taxonomic composition) and estimate their relative abundance in the studied community. However, only a few tools could achieve strain-level identification and most tools estimate the microbial abundances simply according to the read counts. An evaluation study on metagenomic analysis tools concludes that the predicted abundance differed significantly from the true abundance. In this study, we present StrainPro, a novel metagenomic analysis tool which is highly accurate both at characterizing microorganisms at strain-level and estimating their relative abundances. A unique feature of StrainPro is it identifies representative sequence segments from reference genomes. We generate three simulated datasets using known strain sequences and another three simulated datasets using unknown strain sequences. We compare the performance of StrainPro with seven existing tools. The results show that StrainPro not only identifies metagenomes with high precision and recall, but it is also highly robust even when the metagenomes are not included in the reference database. Moreover, StrainPro estimates the relative abundance with high accuracy. We demonstrate that there is a strong positive linear relationship between observed and predicted abundances.

mSystems ◽  
2020 ◽  
Vol 5 (6) ◽  
Author(s):  
Nunzia Picone ◽  
Carmen Hogendoorn ◽  
Geert Cremers ◽  
Lianna Poghosyan ◽  
Arjan Pol ◽  
...  

ABSTRACT Volcanic and geothermal environments are characterized by low pH, high temperatures, and gas emissions consisting of mainly CO2 and varied CH4, H2S, and H2 contents which allow the formation of chemolithoautotrophic microbial communities. To determine the link between the emitted gases and the microbial community composition, geochemical and metagenomic analysis were performed. Soil samples of the geothermic region Favara Grande (Pantelleria, Italy) were taken at various depths (1 to 50 cm). Analysis of the gas composition revealed that CH4 and H2 have the potential to serve as the driving forces for the microbial community. Our metagenomic analysis revealed a high relative abundance of Bacteria in the top layer (1 to 10 cm), but the relative abundance of Archaea increased with depth from 32% to 70%. In particular, a putative hydrogenotrophic methanogenic archaeon, related to Methanocella conradii, appeared to have a high relative abundance (63%) in deeper layers. A variety of [NiFe]-hydrogenase genes were detected, showing that H2 was an important electron donor for microaerobic microorganisms in the upper layers. Furthermore, the bacterial population included verrucomicrobial and proteobacterial methanotrophs, the former showing an up to 7.8 times higher relative abundance. Analysis of the metabolic potential of this microbial community showed a clear capacity to oxidize CH4 aerobically, as several genes for distinct particulate methane monooxygenases and lanthanide-dependent methanol dehydrogenases (XoxF-type) were retrieved. Analysis of the CO2 fixation pathways showed the presence of the Calvin-Benson-Bassham cycle, the Wood-Ljungdahl pathway, and the (reverse) tricarboxylic acid (TCA) cycle, the latter being the most represented carbon fixation pathway. This study indicates that the methane emissions in the Favara Grande might be a combination of geothermal activity and biological processes and further provides insights into the diversity of the microbial population thriving on CH4 and H2. IMPORTANCE The Favara Grande nature reserve on the volcanic island of Pantelleria (Italy) is known for its geothermal gas emissions and high soil temperatures. These volcanic soil ecosystems represent “hot spots” of greenhouse gas emissions. The unique community might be shaped by the hostile conditions in the ecosystem, and it is involved in the cycling of elements such as carbon, hydrogen, sulfur, and nitrogen. Our metagenome study revealed that most of the microorganisms in this extreme environment are only distantly related to cultivated bacteria. The results obtained profoundly increased the understanding of these natural hot spots of greenhouse gas production/degradation and will help to enrich and isolate the microbial key players. After isolation, it will become possible to unravel the molecular mechanisms by which they adapt to extreme (thermo/acidophilic) conditions, and this may lead to new green enzymatic catalysts and technologies for industry.


2019 ◽  
Vol 20 (4) ◽  
pp. 1106-1114 ◽  
Author(s):  
ARTINI PANGASTUTI ◽  
RIZAL KHOIRUN ALFISAH ◽  
NUR IFFAH ISTIANA ◽  
SITI LUSI ARUM SARI ◽  
RATNA SETYANINGSIH ◽  
...  

Abstract. Pangastuti A, Alfisah RK, Istiana NI, Sari SLA, Setyaningsih R, Susilowati A, Purwoko T. 2019. Metagenomic analysis of microbial community in over-fermented tempeh. Biodiversitas 20: 1106-1114. Tempeh is a traditional Indonesian food which is made from soybeans through a fermentation process using Rhizopus as a starter culture. Tempeh is now considered as a functional food with many beneficial effects to human health beyond its nutritional value. The microbial community during the further fermentation process of tempeh give typical characteristic taste and flavor. Therefore the over-fermented tempeh is used as a flavoring in some dishes. Metagenomic analysis needed to know the involvement of microbial communities since most of the microbes involved in further fermentation process are unculturable. This research aimed to study the diversity of the microbial community in the over-fermented tempeh (72 hours) using the metagenomic analysis. Seventeen OTUs of fungi in over-fermented tempeh were detected. Among them, 9 OTUs had significant abundance: six species were identified as Tryblidiopsis sichuanensis, Candida sp.2_1., Kluyveromyces marxianus, Trichosporon asahii, Trichosporon gracile, and Trichosporon ovoides, one species was identified in the order level Mucorales, and two fungi species could not be determined. Species of the order Mucorales was the dominant species in over-fermented tempeh (72 hours) with a relative abundance of 62.46%, followed by Kluyveromyces marxianus with a relative abundance of 3698%. Meanwhile, 132 OTUs of the bacterial community was detected, the predominant 10 genera were Chryseobacterium, Lactococcus, Lactobacillus, Streptococcus, Acetobacter, Novosphingobium, Comamonas, Escherichia, Klebsiella, and Stenotrophomonas, in which Lactobacillus agilis, Lactococcus sp., and Klebsiella sp., were most abundant with relative abundance of 27%, 26.3%, and 13% respectively.


2021 ◽  
pp. 193229682110289
Author(s):  
Evan Olawsky ◽  
Yuan Zhang ◽  
Lynn E Eberly ◽  
Erika S Helgeson ◽  
Lisa S Chow

Background: With the development of continuous glucose monitoring systems (CGMS), detailed glycemic data are now available for analysis. Yet analysis of this data-rich information can be formidable. The power of CGMS-derived data lies in its characterization of glycemic variability. In contrast, many standard glycemic measures like hemoglobin A1c (HbA1c) and self-monitored blood glucose inadequately describe glycemic variability and run the risk of bias toward overreporting hyperglycemia. Methods that adjust for this bias are often overlooked in clinical research due to difficulty of computation and lack of accessible analysis tools. Methods: In response, we have developed a new R package rGV, which calculates a suite of 16 glycemic variability metrics when provided a single individual’s CGM data. rGV is versatile and robust; it is capable of handling data of many formats from many sensor types. We also created a companion R Shiny web app that provides these glycemic variability analysis tools without prior knowledge of R coding. We analyzed the statistical reliability of all the glycemic variability metrics included in rGV and illustrate the clinical utility of rGV by analyzing CGM data from three studies. Results: In subjects without diabetes, greater glycemic variability was associated with higher HbA1c values. In patients with type 2 diabetes mellitus (T2DM), we found that high glucose is the primary driver of glycemic variability. In patients with type 1 diabetes (T1DM), we found that naltrexone use may potentially reduce glycemic variability. Conclusions: We present a new R package and accompanying web app to facilitate quick and easy computation of a suite of glycemic variability metrics.


2021 ◽  
Author(s):  
J. J. Teh ◽  
E. M. Berendsen ◽  
E. C. Hoedt ◽  
S. Kang ◽  
J. Zhang ◽  
...  

AbstractThe mucosa-associated microbiota is widely recognized as a potential trigger for Crohn’s disease pathophysiology but remains largely uncharacterised beyond its taxonomic composition. Unlike stool microbiota, the functional characterisation of these communities using current DNA/RNA sequencing approaches remains constrained by the relatively small microbial density on tissue, and the overwhelming amount of human DNA recovered during sample preparation. Here, we have used a novel ex vivo approach that combines microbe culture from anaerobically preserved tissue with metagenome sequencing (MC-MGS) to reveal patient-specific and strain-level differences among these communities in post-operative Crohn’s disease patients. The 16 S rRNA gene amplicon profiles showed these cultures provide a representative and holistic representation of the mucosa-associated microbiota, and MC-MGS produced both high quality metagenome-assembled genomes of recovered novel bacterial lineages. The MC-MGS approach also produced a strain-level resolution of key Enterobacteriacea and their associated virulence factors and revealed that urease activity underpins a key and diverse metabolic guild in these communities, which was confirmed by culture-based studies with axenic cultures. Collectively, these findings using MC-MGS show that the Crohn’s disease mucosa-associated microbiota possesses taxonomic and functional attributes that are highly individualistic, borne at least in part by novel bacterial lineages not readily isolated or characterised from stool samples using current sequencing approaches.


2021 ◽  
Vol 9 (3) ◽  
pp. 465
Author(s):  
Anne A. M. J. Becker ◽  
KC Hill ◽  
Patrick Butaye

Small Indian mongooses (Urva auropunctata) are among the most pervasive predators to disrupt the native ecology on Caribbean islands and are strongly entrenched in their areas of introduction. Few studies, however, have considered the microbial ecology of such biological invasions. In this study, we investigated the gut microbiota of invasive small Indian mongooses in terms of taxonomic diversity and functional potential. To this end, we collected fecal samples from 60 free-roaming mongooses trapped in different vegetation zones on the island Saint Kitts. The core gut microbiome, assessed by 16S rRNA amplicon gene sequencing on the Ion S5TM XL platform, reflects a carnivore-like signature with a dominant abundance of Firmicutes (54.96%), followed by Proteobacteria (13.98%) and Fusobacteria (12.39%), and a relatively minor contribution of Actinobacteria (10.4%) and Bacteroidetes (6.40%). Mongooses trapped at coastal sites exhibited a higher relative abundance of Fusobacterium spp. whereas those trapped in scrubland areas were enriched in Bacteroidetes, but there was no site-specific difference in predicted metabolic properties. Between males and females, beta-diversity was not significantly different and no sex-specific strategies for energy production were observed. However, the relative abundance of Gammaproteobacteria, and more specifically, Enterobacteriaceae, was significantly higher in males. This first description of the microbial profile of small Indian mongooses provides new insights into their bioecology and can serve as a springboard to further elucidating this invasive predator’s impact throughout the Caribbean.


Agronomy ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1507
Author(s):  
Laura Delgado-Moreno ◽  
Pieter van Dillewijn ◽  
Rogelio Nogales ◽  
Esperanza Romero

The continued discharge of pharmaceuticals and personal care products (PPCPs) into the environment due to their widespread use and the lack of effective systems for their removal from water is a global problem. In this study, the dissipation of ibuprofen, diclofenac and triclosan added simultaneously in biopurification systems (BPSs) with different compositions and their effect on the microbial community structure was analysed. Three BPSs, constituted by mixtures of soil (S), peat (P), or raw wet olive mill cake (A) or its vermicompost (V) and straw (S) were prepared (SPS, SAS and SVS). Sorption and degradation experiments were carried out. After 84 days of incubation, more than 85% of each PPCP applied had dissipated. Methyl-triclosan was determined to be highest in the SVS biomixture. Biomixtures with lower C/N ratio and higher alpha diversity were the most effective in the removal of PPCPs. Initially, the BPS biomixtures showed a different microbial structure dominated by Proteobacteria, Actinobacteria and Bacteroidetes but after addition of PPCPs, a similar pattern was observed in the relative abundance of the phylum Chloroflexi, the class Sphingobacteriia and the genus Brevundimonas. These biopurification systems can be useful to prevent point source contamination due to the disposal of PPCP-contaminated waters.


2021 ◽  
Author(s):  
Qinggeer BORJIGIN ◽  
Bizhou ZHANG ◽  
Xiaofang Yu ◽  
Julin Gao ◽  
Xin ZHANG ◽  
...  

Abstract A lignocellulolytic microbial consortium holds promise for the in situ biodegradation of crop straw and the comprehensive and effective utilization of agricultural waste. In this study, we applied metagenomics technology to comprehensively explore the metabolic functional potential and taxonomic diversity of the microbial consortia CS (cultured on corn stover) and FP (cultured on filter paper).Analyses of the metagenomics taxonomic affiliation data showed considerable differences in the taxonomic composition and functional profile of the microbial consortia CS and FP. The microbial consortia CS primarily contained members from the genera Pseudomonas, Stenotrophomonas, Achromobacter, Dysgonomonas, Flavobacterium and Sphingobacterium, as well as Cellvibrio, Azospirillum, Pseudomonas, Dysgonomonas and Cellulomonas in FP. The COG and KEGG annotation analyses revealed considerable levels of diversity. Further analysis determined that the CS consortium had an increase in the acid and ester metabolism pathways, while carbohydrate metabolism was enriched in the FP consortium. Furthermore, a comparison against the CAZy database showed that the microbial consortia CS and FP contain a rich diversity of lignocellulose degrading families, in which GH5, GH6, GH9, GH10, GH11, GH26, GH42, and GH43 were enriched in the FP consortium, and GH44, GH28, GH2, and GH29 increased in the CS consortium. The degradative mechanism of lignocellulose metabolism by the two microbial consortia is similar, but the annotation of quantity of genes indicated that they are diverse and vary greatly. The lignocellulolytic microbial consortia cultured under different carbon conditions (CS and FP) differed substantially in their composition of the microbial community at the genus level. The changes in functional diversity were accompanied with variation in the composition of microorganisms, many of which are related to the degradation of lignocellulolytic materials. The genera Pseudomonas, Dysgonomonas and Sphingobacterium in CS and the genera Cellvibrio and Pseudomonas in FP exhibited a much wider distribution of lignocellulose degradative ability.


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