Microbial Community Analysis During Start-up of Anaerobic Co-digestion Based on Quinone Profiles Using Supercritical Fluid Extraction

Quinone profile is well known as a useful tool for the analysis of microbial community dynamics in mixed cultures in terms of quantification, simplicity, and reproducibility. The application of quinone profile method in anaerobic digestion is to monitor and overcome instability during fermentation process. A lab-scale anaerobic digestion treating a mixture of milk cow manure (CM) and simulated food waste (FW) during start-up process at mesophilic conditions was used to monitor the change of microbial community dynamics and stability. Supercritical fluid extraction (SFE) experiments using CO2 and ultra-high performance liquid chromatography (UPLC) was applied for extract and determination of ubiquinones (UQ) and menaquinones (MK) species. Quinone can be a helpful tool to make the link between microbial community and anaerobic digestion parameters in order to overcome digester instability during the start-up process

Terminal restriction fragment length polymorphism is an “old school” reliable technique for swift microbial community screening in anaerobic digestion

The microbial community in anaerobic digestion has been analysed through microbial fingerprinting techniques, such as terminal restriction fragment length polymorphism (TRFLP), for decades. In the last decade, high-throughput 16S rRNA gene amplicon sequencing has replaced these techniques, but the time-consuming and complex nature of high-throughput techniques is a potential bottleneck for full-scale anaerobic digestion application, when monitoring community dynamics. Here, the bacterial and archaeal TRFLP profiles were compared with 16S rRNA gene amplicon profiles (Illumina platform) of 25 full-scale anaerobic digestion plants. The α-diversity analysis revealed a higher richness based on Illumina data, compared with the TRFLP data. This coincided with a clear difference in community organisation, Pareto distribution, and co-occurrence network statistics, i.e., betweenness centrality and normalised degree. The β-diversity analysis showed a similar clustering profile for the Illumina, bacterial TRFLP and archaeal TRFLP data, based on different distance measures and independent of phylogenetic identification, with pH and temperature as the two key operational parameters determining microbial community composition. The combined knowledge of temporal dynamics and projected clustering in the β-diversity profile, based on the TRFLP data, distinctly showed that TRFLP is a reliable technique for swift microbial community dynamics screening in full-scale anaerobic digestion plants

Does functional soil microbial diversity contribute to explain within-site plant beta-diversity in an alpine grassland and a <i>dehesa</i> meadow in Spain?

Questions: Once that the effects of hydrological and chemical soil properties have been accounted for, does soil microbial diversity contribute to explain change in plant community structure (i.e. within-site beta-diversity)? If so, at which spatial scale does microbial diversity operate? Location: La Mina in Moscosa Farm, Salamanca, western Spain (dehesa community) and Laguna Larga in the Urbión Peaks, Soria, central-northern Spain (alpine grassland). Methods: The abundance of vascular plant species, soil gram-negative microbial functional types and soil chemical properties (pH, available phosphorus, and extractable cations) were sampled at both sites, for which hydrological models were available. Redundancy analysis (RDA) was used to partition variation in plant community structure into hydrological, chemical and microbial components. Spatial filters, arranged in scalograms, were used to test for the spatial scales at which plant community structure change. Results: In the case of the dehesa the diversity of soil gram-negative microbes, weakly driven by soil pH, contributed to a small extent (adj-R2 = 2%) and at a relative medium spatial scale to explain change in plant community structure. The abundance of a few dehesa species, both annual (Trifolium dubium, Vulpia bromoides) and perennial (Poa bulbosa, Festuca ampla), was associated with either increasing or decreasing soil microbial diversity. In the alpine meadow the contribution was negligible. Conclusions: Microbial diversity can drive community structure, though in the hierarchy of environmental factors structuring communities it appears to rank lower than other soil factors. Still, microbial diversity appears to promote or restrain individual plant species. This paper aims to encourage future studies to use more comprehensive and insightful techniques to assess microbial diversity and to combine this with statistical approaches such as the one used here

Metabolic network analysis reveals microbial community interactions in anammox granules.

Microbial communities mediating anaerobic ammonium oxidation (anammox) represent one of the most energy-efficient environmental biotechnologies for nitrogen removal from wastewater. However, little is known about the functional role heterotrophic bacteria play in anammox granules. Here, we use genome-centric metagenomics to recover 17 draft genomes of anammox and heterotrophic bacteria from a laboratory-scale anammox bioreactor. We combine metabolic network reconstruction with metatranscriptomics to examine the gene expression of anammox and heterotrophic bacteria and to identify their potential interactions. We find that Chlorobi-affiliated bacteria may be highly active protein degraders, catabolizing extracellular peptides while recycling nitrate to nitrite. Other heterotrophs may also contribute to scavenging of detritus and peptides produced by anammox bacteria, and potentially use alternative electron donors, such as H2, acetate and formate. Our findings improve the understanding of metabolic activities and interactions between anammox and heterotrophic bacteria and offer the first transcriptional insights on ecosystem function in anammox granules

Changes in the genetic requirements for microbial interactions with increasing community complexity.

Microbial community structure and function rely on complex interactions whose underlying molecular mechanisms are poorly understood. To investigate these interactions in a simple microbiome, we introduced E. coli into an experimental community based on a cheese rind and identified the differences in E. coli's genetic requirements for growth in interactive and non-interactive contexts using Random Barcode Transposon Sequencing (RB-TnSeq) and RNASeq. Genetic requirements varied among pairwise growth conditions and between pairwise and community conditions. Our analysis points to mechanisms by which growth conditions change as a result of increasing community complexity and suggests that growth within a community relies on a combination of pairwise and higher-order interactions. Our work provides a framework for using the model organism E. coli as a readout to investigate microbial interactions regardless of the genetic tractability of members of the studied ecosystem

Nitrogen removal in a two-chambered microbial fuel cell: Establishment of a nitrifying-denitrifying microbial community on an intermittent aerated cathode

A microbial fuel cell (MFC) was used to study nitrogen dynamics and its feasibility for high strength wastewater treatment. Intermittent aeration was applied on the cathode chamber accomplishing the establishment of a simultaneous nitrifying-denitrifying microbial community. A total of 30.4% of the N-NH4 + migrated through the ion exchange membrane being primarily nitrified at the cathode chamber. When intermittent aeration was applied in the cathode, denitrification also occurred achieving 17.8% of nitrate removal without acetate addition, and 41.2% with acetate addition. The microbial community analysis revealed that the nitrification process at the cathode chamber could be explained due to a high predominance of Nitrosomonas sp. as ammonia-oxidising bacteria and other Comamonadaceae phylotypes as potential denitrifiers. Parallel batch denitrification assays, carried out outside the MFC using the cathode effluent, confirmed the existence of heterotrophic denitrification processes with other well known denitrifying dominant phylotypes enrichment (Burkholderiadaceae, Comamonadaceae, Alcaligenaceae).Postprint (author's final draft

Microbial residence time is a controlling parameter of the taxonomic composition and functional profile of microbial communities.

A remaining challenge within microbial ecology is to understand the determinants of richness and diversity observed in environmental microbial communities. In a range of systems, including activated sludge bioreactors, the microbial residence time (MRT) has been previously shown to shape the microbial community composition. However, the physiological and ecological mechanisms driving this influence have remained unclear. Here, this relationship is explored by analyzing an activated sludge system fed with municipal wastewater. Using a model designed in this study based on Monod-growth kinetics, longer MRTs were shown to increase the range of growth parameters that enable persistence, resulting in increased richness and diversity in the modeled community. In laboratory experiments, six sequencing batch reactors treating domestic wastewater were operated in parallel at MRTs between 1 and 15 days. The communities were characterized using both 16S ribosomal RNA and non-target messenger RNA sequencing (metatranscriptomic analysis), and model-predicted monotonic increases in richness were confirmed in both profiles. Accordingly, taxonomic Shannon diversity also increased with MRT. In contrast, the diversity in enzyme class annotations resulting from the metatranscriptomic analysis displayed a non-monotonic trend over the MRT gradient. Disproportionately high abundances of transcripts encoding for rarer enzymes occur at longer MRTs and lead to the disconnect between taxonomic and functional diversity profiles

Enteropathogen survival in soil from different land-uses is predominantly regulated by microbial community composition

peer-reviewedMicrobial enteropathogens can enter the environment via landspreading of animal slurries and manures. Biotic interactions with the soil microbial community can contribute to their subsequent decay. This study aimed to determine the relative impact of biotic, specifically microbial community structure, and physico-chemical properties associated with soils derived from 12 contrasting land-uses on enteropathogen survival. Phenotypic profiles of microbial communities (via phospholipid fatty acid (PLFA) profiling), and total biomass (by fumigation-extraction), in the soils were determined, as well as a range of physicochemical properties. The persistence of Salmonella Dublin, Listeria monocytogenes, and Escherichia coli was measured over 110 days within soil microcosms. Physicochemical and biotic data were used in stepwise regression analysis to determine the predominant factor related to pathogen-specific death rates. Phenotypic structure, associated with a diverse range of constituent PLFAs, was identified as the most significant factor in pathogen decay for S. Dublin, L. monocytogenes, non-toxigenic E. coli O157 but not for environmentally-persistent E. coli. This demonstrates the importance of entire community-scale interactions in pathogen suppression, and that such interactions are context-specific

Microbial Community Analysis During Start-up of Anaerobic Co-digestion Based on Quinone Profiles Using Supercritical Fluid Extraction

Quinone profile is well known as a useful tool for the analysis of microbial community dynamics in mixed cultures in terms of quantification, simplicity, and reproducibility. The application of quinone profile method in anaerobic digestion is to monitor and overcome instability during fermentation process. A lab-scale anaerobic digestion treating a mixture of milk cow manure (CM) and simulated food waste (FW) during start-up process at mesophilic conditions was used to monitor the change of microbial community dynamics and stability. Supercritical fluid extraction (SFE) experiments using CO2 and ultra-high performance liquid chromatography (UPLC) was applied for extract and determination of ubiquinones (UQ) and menaquinones (MK) species. Quinone can be a helpful tool to make the link between microbial community and anaerobic digestion parameters in order to overcome digester instability during the start-up process

Microbial differences between dental plaque and historic dental calculus are related to oral biofilm maturation stage

Dental calculus, calcified oral plaque biofilm, contains microbial and host biomolecules that can be used to study historic microbiome communities and host responses. Dental calculus does not typically accumulate as much today as historically, and clinical oral microbiome research studies focus primarily on living dental plaque biofilm. However, plaque and calculus reflect different conditions of the oral biofilm, and the differences in microbial characteristics between the sample types have not yet been systematically explored. Here, we compare the microbial profiles of modern dental plaque, modern dental calculus, and historic dental calculus to establish expected differences between these substrates.- Background - Results -- Authentication of a preserved oral biofilm in calculus samples -- Dental calculus and plaque biofilm communities are distinct -- Health-associated communities of dental plaque and calculus are distinct -- Signatures of health and of disease are shared in modern and historic calculus samples -- Microbial community differences between health and disease in calculus are poorly resolved -- Absence of caries-specific microbial profiles in dental calculus -- Microbial co-exclusion patterns in plaque and calculus reflect biofilm maturity -- Microbial complexes in plaque and calculus -- Functional prediction in calculus is poorly predictive of health status -- Proteomic profiles of historic healthy site calculus -- Correlations between taxonomic, proteomic, and metabolomic profiles - Discussion - Conclusions - Materials and methods --Historic and modern calculus sample collection DNA extraction -- DNA library construction and high-throughput sequencing -- DNA sequence processing -- Genetic assessment of historic calculus sample preservation -- Genetic microbial taxonomic profiling -- Principal component analysis -- Assessment of differentially abundant taxa -- Sparse partial least squares-discriminant analysis -- Assessment of microbial co-exclusion patterns -- Gene functional categorization with SEED -- Proteomics -- Metabolomics -- Regularized canonical correlation analysi