scholarly journals Refinement of metabolite detection in cystic fibrosis sputum reveals heme negatively correlates with lung function

2018 ◽  
Author(s):  
Nathaniel R. Glasser ◽  
Ryan C. Hunter ◽  
Theodore G. Liou ◽  
Dianne K. Newman ◽  
Keyword(s):  
Mass Spectrometry ◽  
Cystic Fibrosis ◽  
Lung Function ◽  
High Performance ◽  
16S Rrna Gene Sequencing ◽  
Initial Study ◽  
Rrna Gene ◽  
Sequencing Data ◽  
Unknown Compound ◽  
Mountain West

SummaryPseudomonas aeruginosalung infections are a leading cause of morbidity and mortality in cystic fibrosis (CF) patients (1, 2). Our laboratory has studied a class of small molecules produced byP. aeruginosaknown as phenazines, including pyocyanin and its biogenic precursor phenazine-1-carboxylic acid (PCA). As phenazines are known virulence factors (3), we and others have explored the possibility of using phenazine concentrations as a marker for disease progression (4–6). Previously, we reported that sputum concentrations of pyocyanin and PCA negatively correlate with lung function in cystic fibrosis patients (6). Our study used high performance liquid chromatography (HPLC) to quantify phenazines by UV–vis absorbance after extraction from lung sputum. Since our initial study, methods for metabolite analysis have advanced considerably, aided in large part by usage of mass spectrometry (LC-MS) and tandem mass spectrometry (LC-MS/MS). Because a more recent study employing LC-MS/MS revealed a surprising decoupling ofP. aeruginosametabolites in sputum and the detection ofP. aeruginosathrough culturing or microbiome profiles (4), we decided to check whether we could reproduce our previous findings by analyzing sputum samples from a different patient cohort with a new LC-MS instrument in our laboratory. Our new samples were provided by the Mountain West CF Consortium Sputum Biomarker study (7). In the course of performing our new analyses, comparison of our old HPLC data to our new LC-MS data led us to realize that the peak previously assigned to PCA instead originates from heme, and the peak assigned to pyocyanin originates from an as-yet unknown compound. This correction only affects the measurements of phenazines in sputum, and we are confident in the phenazine measurements from isolated cultures and the 16S rRNA gene sequencing data from that study (6). Here we outline the basis for our correction and present additional data showing that heme concentration negatively correlates with lung function in cystic fibrosis patients.

scholarly journals A Volatile and Dynamic Longitudinal Microbiome Is Associated With Less Reduction in Lung Function in Adolescents With Cystic Fibrosis

2021 ◽  
Vol 11 ◽  
Author(s):  
Marisa I. Metzger ◽  
Simon Y. Graeber ◽  
Mirjam Stahl ◽  
Olaf Sommerburg ◽  
Marcus A. Mall ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Lung Function ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Contributing Factors ◽  
Lung Function Decline ◽  
Cross Sectional ◽  
Lung Microbiome ◽  
The Stability ◽  
The Individual

Progressive impairment in lung function caused by chronic polymicrobial airway infection remains the major cause of death in patients with cystic fibrosis (CF). Cross-sectional studies suggest an association between lung function decline and specific lung microbiome ecotypes. However, longitudinal studies on the stability of the airway microbiome are missing for adolescents with CF constituting the age group showing the highest rate of decline in lung function. In this study, we analyzed longitudinal lung function data and sputum samples collected over a period of 3 to 5 years from 12 adolescents with CF. The sputum microbiome was analyzed using 16S rRNA gene sequencing. Our results indicate that the individual course of the lung microbiome is associated with longitudinal lung function. In our cohort, patients with a dynamic, diverse microbiome showed a slower decline of lung function measured by FEV1% predicted, whereas a more stable and less diverse lung microbiome was related to worse outcomes. Specifically, a higher abundance of the phyla Bacteroidetes and Firmicutes was linked to a better clinical outcome, while Proteobacteria were correlated with a decline in FEV1% predicted. Our study indicates that the stability and diversity of the lung microbiome and the abundance of Bacteroidetes and Firmicutes are associated with the lung function decline and are one of the contributing factors to the disease severity.

scholarly journals Manganese/iron-supported sulfate-dependent anaerobic oxidation of methane by archaea in lake sediments

2019 ◽  
Author(s):  
Guangyi Su ◽  
Jakob Zopfi ◽  
Haoyi Yao ◽  
Lea Steinle ◽  
Helge Niemann ◽  
...  
Keyword(s):  
Lake Sediments ◽  
16S Rrna Gene Sequencing ◽  
Electron Acceptors ◽  
Rrna Gene ◽  
Anaerobic Oxidation Of Methane ◽  
Anaerobic Oxidation ◽  
Sequencing Data ◽  
Oxidation Of Methane ◽  
Freshwater Habitats ◽  
Rrna Gene Sequencing

AbstractAnaerobic oxidation of methane (AOM) by methanotrophic archaea is an important sink of this greenhouse gas in marine sediments. However, evidence for AOM in freshwater habitats is rare, and little is known about the pathways, electron acceptors and microbes involved. Here, we show that AOM occurs in anoxic sediments of a lake in southern Switzerland (Lake Cadagno). Combined AOM-rate and 16S rRNA gene-sequencing data suggest thatCandidatusMethanoperedens archaea are responsible for the observed methane oxidation. Members of the Methanoperedenaceae family were previously reported to conduct nitrate- or iron/manganese-dependent AOM. However, we demonstrate for the first time that the methanotrophic archaea do not necessarily rely upon these oxidants as terminal electron acceptors directly, but mainly perform canonical sulfate-dependent AOM, which under sulfate-starved conditions can be supported by metal (Mn, Fe) oxides through oxidation of reduced sulfur species to sulfate. The correspondence of high abundances of Desulfobulbaceae andCandidatusMethanoperedens at the same sediment depth confirm the interdependence of anaerobic methane-oxidizing archaea and sulfate-reducing bacteria. The relatively high abundance and widespread distribution ofCandidatusMethanoperedens in lake sediments highlight their potentially important role in mitigating methane emissions from terrestrial freshwater environments to the atmosphere, analogous to ANME-1, -2 and -3 in marine settings.

scholarly journals Dynamics of Soil Microbial Communities During Diazepam and Oxazepam Biodegradation in Soil Flooded by Water From a WWTP

2021 ◽  
Vol 12 ◽  
Author(s):  
Marc Crampon ◽  
Coralie Soulier ◽  
Pauline Sidoli ◽  
Jennifer Hellal ◽  
Catherine Joulian ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Microbial Communities ◽  
Soil Microbial Communities ◽  
Gene Sequencing ◽  
16S Rrna Gene Sequencing ◽  
Treated Wastewater ◽  
Rrna Gene ◽  
Sequencing Data ◽  
Rrna Gene Sequencing

The demand for energy and chemicals is constantly growing, leading to an increase of the amounts of contaminants discharged to the environment. Among these, pharmaceutical molecules are frequently found in treated wastewater that is discharged into superficial waters. Indeed, wastewater treatment plants (WWTPs) are designed to remove organic pollution from urban effluents but are not specific, especially toward contaminants of emerging concern (CECs), which finally reach the natural environment. In this context, it is important to study the fate of micropollutants, especially in a soil aquifer treatment (SAT) context for water from WWTPs, and for the most persistent molecules such as benzodiazepines. In the present study, soils sampled in a reed bed frequently flooded by water from a WWTP were spiked with diazepam and oxazepam in microcosms, and their concentrations were monitored for 97 days. It appeared that the two molecules were completely degraded after 15 days of incubation. Samples were collected during the experiment in order to follow the dynamics of the microbial communities, based on 16S rRNA gene sequencing for Archaea and Bacteria, and ITS2 gene for Fungi. The evolution of diversity and of specific operating taxonomic units (OTUs) highlighted an impact of the addition of benzodiazepines, a rapid resilience of the fungal community and an evolution of the bacterial community. It appeared that OTUs from the Brevibacillus genus were more abundant at the beginning of the biodegradation process, for diazepam and oxazepam conditions. Additionally, Tax4Fun tool was applied to 16S rRNA gene sequencing data to infer on the evolution of specific metabolic functions during biodegradation. It finally appeared that the microbial community in soils frequently exposed to water from WWTP, potentially containing CECs such as diazepam and oxazepam, may be adapted to the degradation of persistent contaminants.

scholarly journals A pilot study of the effect of deployment on the gut microbiome and traveler’s diarrhea susceptibility

2020 ◽  
Author(s):  
Blake W. Stamps ◽  
Wanda J. Lyon ◽  
Adam P. Irvin ◽  
Nancy Kelley-Loughnane ◽  
Michael S. Goodson
Keyword(s):  
Pilot Study ◽  
16S Rrna ◽  
Gut Microbiome ◽  
16S Rrna Gene Sequencing ◽  
Initial Study ◽  
Mitigation Strategies ◽  
Taxonomic Resolution ◽  
Rrna Gene ◽  
Traveler’S Diarrhea ◽  
Traveler's Diarrhea

AbstractTraveler’s diarrhea (TD) is a recurrent and significant issue for many travelers including the military. While many known enteric pathogens exist that are causative agents of diarrhea, our gut microbiome may also play a role in travelers’ diarrhea susceptibility. To this end we conducted a pilot study of the microbiome of warfighters prior to- and after deployment overseas to identify marker taxa relevant to traveler’s diarrhea. This initial study utilized full-length 16S rRNA gene sequencing to provide additional taxonomic resolution towards identifying predictive taxa.16S rRNA analyses of pre- and post-deployment fecal samples identified multiple marker taxa as significantly differentially abundant in subjects that reported diarrhea, including Weissella, Butyrivibrio, Corynebacterium, uncultivated Erysipelotrichaceae, Jeotgallibaca, unclassified Ktedonobacteriaceae, Leptolinea, and uncultivated Ruminiococcaceae. The ability to identify TD risk prior to travel will inform prevention and mitigation strategies to influence diarrhea susceptibility while traveling.

199 Do the Microbiomes of Horses Process Nutrients Differently Based on Keeper Status?

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 106-106
Author(s):  
Alexa C Johnson ◽  
Amy S Biddle
Keyword(s):  
Fixed Effects ◽  
High Performance ◽  
16S Rrna Gene Sequencing ◽  
Gas Production ◽  
Rrna Gene ◽  
Total N ◽  
Fermentation Products ◽  
Fecal Samples ◽  
Α Diversity ◽  
C 50

Abstract This study reports the differential response of the equine gut microbiome to protein and/or carbohydrate based on keeper status (easy keeper (EK), medium keeper (MK), hard keeper (HK)). Anaerobic equine fecal samples (n = 12 total, n = 3 / EK, MK, HK of four breeds) inoculated microcosms with three dietary conditions (C = Carb (cornmeal), P = Protein (soybean meal), and M = mix (50% C, 50% P)). Over 48 hours, fermentation products were measured using colorimetric assays and high-performance liquid chromatography. Microbial populations were surveyed using 16S rRNA gene sequencing analyzed by QIIME2. Linear mixed models were fit with fixed effects of Treatment and Keeper status and their interactions, with random effects of HorseID. Differences in fermentation products by keeper status included: MK had higher pH and greater gas production, EK produced higher hydrogen sulfide, and HK had greater total protein. Total SCFA was not different between keeper status (P = 0.89) but the acetate: propionate ratio was highest for HK (2.45mM) and lowest for EK (1.85mM) (P = 0.05). Isobutyrate production was highest in HK (2.34mM) compared to MK (0.85mM) and EK (0.17mM). Treatment had significant effects across all measurements; M and C treatment values were similar reflecting microbial preferences for carbohydrates before protein. P treated trials had increased fermentation outputs due to lower acidity effects. Keeper status had no effect on α-diversity (P > 0.05) however HK horses were least affected by treatments. P treated samples were more diverse than C and M (P < 0.001). Spearman correlation of Keeper x Treatment identified Oligosphaeria spp. in EK (r = 0.49) and Fusobacteria spp. in HK whole fecal samples (r = 0.37). These data suggest that while the compositions of the gut microbiomes of keeper groups were similar, they were functionally different in processing key nutrients.

scholarly journals Identification of a Shared Microbiomic and Metabolomic Profile in Systemic Autoimmune Diseases

2019 ◽  
Vol 8 (9) ◽  
pp. 1291 ◽  
Author(s):  
Bellocchi ◽  
Fernández-Ochoa ◽  
Montanelli ◽  
Vigone ◽  
Santaniello ◽  
...  
Keyword(s):  
Autoimmune Diseases ◽  
Lupus Erythematosus ◽  
High Performance ◽  
16S Rrna Gene Sequencing ◽  
Study Groups ◽  
Mass Spectrometry Analysis ◽  
Rrna Gene ◽  
Operating Characteristics ◽  
Systemic Autoimmune Diseases ◽  
Spectrometry Analysis

Dysbiosis has been described in systemic autoimmune diseases (SADs), including systemic lupus erythematosus (SLE), Sjögren’s syndrome (SjS), and primary anti-phosholipid syndrome (PAPS), however the biological implications of these associations are often elusive. Stool and plasma samples from 114 subjects, including in SLE (n = 27), SjS (n = 23), PAPs (n = 11) and undifferentiated connective tissue (UCTD, n = 26) patients, and geographically-matched healthy controls (HCs, n = 27), were collected for microbiome (16s rRNA gene sequencing) and metabolome (high-performance liquid chromatography coupled to mass spectrometry) analysis to identify shared characteristics across diseases. Out of 130 identified microbial genera, a subset of 29 bacteria was able to differentiate study groups (area under receiver operating characteristics (AUROC) = 0.730 ± 0.025). A fair classification was obtained with a subset of 41 metabolic peaks out of 254 (AUROC = 0.748 ± 0.021). In both models, HCs were well separated from SADs, while UCTD largely overlapped with the other diseases. In all of the SADs pro-tolerogenic bacteria were reduced, while pathobiont genera were increased. Metabolic alterations included two clusters comprised of: (a) members of the acylcarnitine family, positively correlating with a Prevotella-enriched cluster and negatively correlating with a butyrate-producing bacteria-enriched cluster; and (b) phospholipids, negatively correlating with butyrate-producing bacteria. These findings demonstrate a strong interaction between intestinal microbiota and metabolic function in patients with SADs.

scholarly journals Hierarchical Microbial Functions Prediction by Graph Aggregated Embedding

2021 ◽  
Vol 11 ◽  
Author(s):  
Yujie Hou ◽  
Xiong Zhang ◽  
Qinyan Zhou ◽  
Wenxing Hong ◽  
Ying Wang
Keyword(s):  
16S Rrna ◽  
Penaeus Monodon ◽  
Poor Performance ◽  
16S Rrna Gene Sequencing ◽  
Reference Database ◽  
Rrna Gene ◽  
Sequencing Data ◽  
Significant Generalization ◽  
Rrna Gene Sequencing ◽  
Almost All

Matching 16S rRNA gene sequencing data to a metabolic reference database is a meaningful way to predict the metabolic function of bacteria and archaea, bringing greater insight to the working of the microbial community. However, some operational taxonomy units (OTUs) cannot be functionally profiled, especially for microbial communities from non-human samples cultured in defective media. Therefore, we herein report the development of Hierarchical micrObial functions Prediction by graph aggregated Embedding (HOPE), which utilizes co-occurring patterns and nucleotide sequences to predict microbial functions. HOPE integrates topological structures of microbial co-occurrence networks with k-mer compositions of OTU sequences and embeds them into a lower-dimensional continuous latent space, while maximally preserving topological relationships among OTUs. The high imbalance among KEGG Orthology (KO) functions of microbes is recognized in our framework that usually yields poor performance. A hierarchical multitask learning module is used in HOPE to alleviate the challenge brought by the long-tailed distribution among classes. To test the performance of HOPE, we compare it with HOPE-one, HOPE-seq, and GraphSAGE, respectively, in three microbial metagenomic 16s rRNA sequencing datasets, including abalone gut, human gut, and gut of Penaeus monodon. Experiments demonstrate that HOPE outperforms baselines on almost all indexes in all experiments. Furthermore, HOPE reveals significant generalization ability. HOPE's basic idea is suitable for other related scenarios, such as the prediction of gene function based on gene co-expression networks. The source code of HOPE is freely available at https://github.com/adrift00/HOPE.

scholarly journals Nonmarine Crenarchaeol in Nevada Hot Springs

2004 ◽  
Vol 70 (9) ◽  
pp. 5229-5237 ◽  
Author(s):  
A. Pearson ◽  
Z. Huang ◽  
A. E. Ingalls ◽  
C. S. Romanek ◽  
J. Wiegel ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Performance ◽  
Hot Springs ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Membrane Lipids ◽  
Marine Species ◽  
Gas Chromatography Mass Spectrometry ◽  
Rrna Gene ◽  
Chromatography Mass Spectrometry ◽  
Group I

ABSTRACT Glycerol dialkyl glycerol tetraethers (GDGTs) are core membrane lipids of the Crenarchaeota. The structurally unusual GDGT crenarchaeol has been proposed as a taxonomically specific biomarker for the marine planktonic group I archaea. It is found ubiquitously in the marine water column and in sediments. In this work, samples of microbial community biomass were obtained from several alkaline and neutral-pH hot springs in Nevada, United States. Lipid extracts of these samples were analyzed by high-performance liquid chromatography-mass spectrometry and by gas chromatography-mass spectrometry. Each sample contained GDGTs, and among these compounds was crenarchaeol. The distribution of archaeal lipids in Nevada hot springs did not appear to correlate with temperature, as has been observed in the marine environment. Instead, a significant correlation with the concentration of bicarbonate was observed. Archaeal DNA was analyzed by denaturing gradient gel electrophoresis. All samples contained 16S rRNA gene sequences which were more strongly related to thermophilic crenarchaeota than to Cenarchaeum symbiosum, a marine nonthermophilic crenarchaeon. The occurrence of crenarchaeol in environments containing sequences affiliated with thermophilic crenarchaeota suggests a wide phenotypic distribution of this compound. The results also indicate that crenarchaeol can no longer be considered an exclusive biomarker for marine species.

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