Dataset Information

Identifying Eukaryotes and Factors Influencing Their Biogeography in Drinking Water Metagenomes.

ABSTRACT: The biogeography of eukaryotes in drinking water systems is poorly understood relative to that of prokaryotes or viruses, limiting the understanding of their role and management. A challenge with studying complex eukaryotic communities is that metagenomic analysis workflows are currently not as mature as those that focus on prokaryotes or viruses. In this study, we benchmarked different strategies to recover eukaryotic sequences and genomes from metagenomic data and applied the best-performing workflow to explore the factors affecting the relative abundance and diversity of eukaryotic communities in drinking water distribution systems (DWDSs). We developed an ensemble approach exploiting k-mer- and reference-based strategies to improve eukaryotic sequence identification and identified MetaBAT2 as the best-performing binning approach for their clustering. Applying this workflow to the DWDS metagenomes showed that eukaryotic sequences typically constituted small proportions (i.e., <1%) of the overall metagenomic data with higher relative abundances in surface water-fed or chlorinated systems with high residuals. The α and β diversities of eukaryotes were correlated with those of prokaryotic and viral communities, highlighting the common role of environmental/management factors. Finally, a co-occurrence analysis highlighted clusters of eukaryotes whose members' presence and abundance in DWDSs were affected by disinfection strategies, climate conditions, and source water types.

SUBMITTER: Gabrielli M

PROVIDER: S-EPMC9996835 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature

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Identifying Eukaryotes and Factors Influencing Their Biogeography in Drinking Water Metagenomes.

Gabrielli Marco M Dai Zihan Z Delafont Vincent V Timmers Peer H A PHA van der Wielen Paul W J J PWJJ Antonelli Manuela M Pinto Ameet J AJ

Environmental science & technology 20230224 9

The biogeography of eukaryotes in drinking water systems is poorly understood relative to that of prokaryotes or viruses, limiting the understanding of their role and management. A challenge with studying complex eukaryotic communities is that metagenomic analysis workflows are currently not as mature as those that focus on prokaryotes or viruses. In this study, we benchmarked different strategies to recover eukaryotic sequences and genomes from metagenomic data and applied the best-performing w ...[more]

PMID: 36827617

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Project description:BACKGROUND:Eukaryotes are ubiquitous in natural environments such as soil and freshwater. Little is known of their presence in drinking water distribution systems (DWDSs) or of the environmental conditions that affect their activity and survival. METHODS:Eukaryotes were characterized by Illumina high-throughput sequencing targeting 18S rRNA gene (DNA) that estimates the total community and the 18S rRNA gene transcript (RNA) that is more representative of the active part of the community. DWDS cold water (N = 124), hot water (N = 40), and biofilm (N = 16) samples were collected from four cities in Finland. The sampled DWDSs were from two waterworks A-B with non-disinfected, recharged groundwater as source water and from three waterworks utilizing chlorinated water (two DWDSs of surface waterworks C-D and one of ground waterworks E). In each DWDS, samples were collected from three locations during four seasons of 1 year. RESULTS:A beta-diversity analysis revealed that the main driver shaping the eukaryotic communities was the DWDS (A-E) (R = 0.73, P < 0.001, ANOSIM). The kingdoms Chloroplastida (green plants and algae), Metazoa (animals: rotifers, nematodes), Fungi (e.g., Cryptomycota), Alveolata (ciliates, dinoflagellates), and Stramenopiles (algae Ochrophyta) were well represented and active-judging based on the rRNA gene transcripts-depending on the surrounding conditions. The unchlorinated cold water of systems (A-B) contained a higher estimated total number of taxa (Chao1, average 380-480) than chlorinated cold water in systems C-E (Chao1 ≤ 210). Within each DWDS, unique eukaryotic communities were identified at different locations as was the case also for cold water, hot water, and biofilms. A season did not have a consistent impact on the eukaryotic community among DWDSs. CONCLUSIONS:This study comprehensively characterized the eukaryotic community members within the DWDS of well-maintained ground and surface waterworks providing good quality water. The study gives an indication that each DWDS houses a unique eukaryotic community, mainly dependent on the raw water source and water treatment processes in place at the corresponding waterworks. In particular, disinfection as well as hot water temperature seemed to represent a strong selection pressure that controlled the number of active eukaryotic species.

Project description:Reconstructing microbial genomes from metagenomic short-read data can be challenging due to the unknown and uneven complexity of microbial communities. This complexity encompasses highly diverse populations, which often includes strain variants. Reconstructing high-quality genomes is a crucial part of the metagenomic workflow, as subsequent ecological and metabolic inferences depend on their accuracy, quality, and completeness. In contrast to microbial communities in other ecosystems, there has been no systematic assessment of genome-centric metagenomic workflows for drinking water microbiomes. In this study, we assessed the performance of a combination of assembly and binning strategies for time series drinking water metagenomes that were collected over 6 months. The goal of this study was to identify the combination of assembly and binning approaches that result in high-quality and -quantity metagenome-assembled genomes (MAGs), representing most of the sequenced metagenome. Our findings suggest that the metaSPAdes coassembly strategies had the best performance, as they resulted in larger and less fragmented assemblies, with at least 85% of the sequence data mapping to contigs greater than 1 kbp. Furthermore, a combination of metaSPAdes coassembly strategies and MetaBAT2 produced the highest number of medium-quality MAGs while capturing at least 70% of the metagenomes based on read recruitment. Utilizing different assembly/binning approaches also assists in the reconstruction of unique MAGs from closely related species that would have otherwise collapsed into a single MAG using a single workflow. Overall, our study suggests that leveraging multiple binning approaches with different metaSPAdes coassembly strategies may be required to maximize the recovery of good-quality MAGs. IMPORTANCE Drinking water contains phylogenetic diverse groups of bacteria, archaea, and eukarya that affect the esthetic quality of water, water infrastructure, and public health. Taxonomic, metabolic, and ecological inferences of the drinking water microbiome depend on the accuracy, quality, and completeness of genomes that are reconstructed through the application of genome-resolved metagenomics. Using time series metagenomic data, we present reproducible genome-centric metagenomic workflows that result in high-quality and -quantity genomes, which more accurately signifies the sequenced drinking water microbiome. These genome-centric metagenomic workflows will allow for improved taxonomic and functional potential analysis that offers enhanced insights into the stability and dynamics of drinking water microbial communities.

Project description:In deep drinking water reservoir ecosystems, the dynamics and interactions of community compositions of phytoplankton and eukaryotes during the mixing periods are still unclear. Here, morphological characteristics combined with high-throughput DNA sequencing (HTS) were used to investigate the variations of phytoplankton and the eukaryotic community in a large canyon-shaped, stratified reservoir located at the Heihe River in Shaanxi Province for three months. The results showed that Bacillariophyta and Chlorophyta were the dominant taxa of the phytoplankton community, accounting for more than 97% of total phytoplankton abundance, which mainly consisted of Melosira sp., Cyclotella sp., and Chlorella sp., respectively. Illumina Miseq sequencing suggested that the biodiversity of eukaryotes increased over time and thatspecies distribution was more even. Arthropoda (6.63% to 79.19%), Ochrophyta (5.60% to 35.16%), Ciliophora (1.81% to 10.93%) and Cryptomonadales (0.25% to 11.48%) were the keystone taxa in common, contributing over 50% of the total eukaryotic community. Cryptomycota as a unique fungus was observed to possess significant synchronization with algal density, reaching a maximum of 10.70% in December (when the algal density distinctly decreased) and suggesting that it might affect the growth of algae through parasitism. Co-occurrence network patterns revealed the complicated and diverse interactions between eukaryotes and phytoplankton, suggesting that eukaryotes respond to variations in dynamic structure of the phytoplankton community, although there might be antagonistic or mutualistic interactions between them. Redundancy analysis (RDA) results showed that environmental variables collectively explained a 96.7% variance of phytoplankton and 96.3% variance of eukaryotic microorganisms, indicating that the temporal variations of phytoplankton and eukaryotic microorganisms were significantly affected by environmental conditions. This study shows that potential interactions exist between phytoplankton and eukaryotic microorganism communities, andcould improve our understanding of the ecological roles of phytoplankton and eukaryotic microorganisms in changing aquatic ecosystems. However, long-term investigations are necessary in order to obtain comprehensive understandings of their complicated associations.

Project description:IntroductionColorectal cancer is the second deadliest cancer worldwide. One of the risk factors for the development of this type of cancer is alcohol consumption. Patients with colorectal cancer may be stigmatized regarding their cancer and regarding drinking behaviors they may exhibit. This study aimed to analyze community persons' and health professionals' acceptability judgments regarding alcohol drinkers having colorectal cancer.MethodThis study relies on an experimental method enabling the identification of variables involved in one's judgment, based on the exhaustive combination of factors yielding several scenarios rated by participants. Scenarios implemented factors possibly influencing participants' perception of a woman character having colorectal cancer. Factors included her drinking habits, post-diagnosis drinking behavior and type of diagnosis/prognosis. The participants were community persons (N' = 132) or health professionals (N" = 126). Data were analyzed using a within-subject factorial ANOVA.ResultsIn both samples, the "Post-diagnosis behavior" factor had large effect sizes, with drinking cessation being more acceptable than other drinking behaviors. Another factor, "Drinking habits", had significant influences on participants judgments, as higher drinking was considered less acceptable. A third factor, "Diagnosis" (polyps, early- or late-stage cancer), was taken into account by participants when it interacted with "Drinking habits" and "Post-diagnosis behavior". Indeed, participants considered most acceptable to continue drinking in the case of late-stage cancer, especially in the health professional sample where the acceptability of continuing drinking was almost doubled when the character had advanced- rather than early-cancer.ConclusionThe lesser the drinking behavior, the better the acceptability. However, advanced cancer stage attenuated the poor acceptability of drinking in both samples, as participants' attitudes were more permissive when the patient had advanced cancer.

Project description:Biological stability of drinking water refers to the concept of providing consumers with drinking water of same microbial quality at the tap as produced at the water treatment facility. However, uncontrolled growth of bacteria can occur during distribution in water mains and premise plumbing, and can lead to hygienic (e.g., development of opportunistic pathogens), aesthetic (e.g., deterioration of taste, odor, color) or operational (e.g., fouling or biocorrosion of pipes) problems. Drinking water contains diverse microorganisms competing for limited available nutrients for growth. Bacterial growth and interactions are regulated by factors, such as (i) type and concentration of available organic and inorganic nutrients, (ii) type and concentration of residual disinfectant, (iii) presence of predators, such as protozoa and invertebrates, (iv) environmental conditions, such as water temperature, and (v) spatial location of microorganisms (bulk water, sediment, or biofilm). Water treatment and distribution conditions in water mains and premise plumbing affect each of these factors and shape bacterial community characteristics (abundance, composition, viability) in distribution systems. Improved understanding of bacterial interactions in distribution systems and of environmental conditions impact is needed for better control of bacterial communities during drinking water production and distribution. This article reviews (i) existing knowledge on biological stability controlling factors and (ii) how these factors are affected by drinking water production and distribution conditions. In addition, (iii) the concept of biological stability is discussed in light of experience with well-established and new analytical methods, enabling high throughput analysis and in-depth characterization of bacterial communities in drinking water. We discussed, how knowledge gained from novel techniques will improve design and monitoring of water treatment and distribution systems in order to maintain good drinking water microbial quality up to consumer's tap. A new definition and methodological approach for biological stability is proposed.

Project description:Concentration- and time-dependent genomic changes in the mouse urinary bladder following exposure to arsenate in drinking water for up to twelve weeks. Inorganic arsenic (Asi) is a known human bladder carcinogen. The objective of this study was to examine the concentration dependence of the genomic response to Asi in the urinary bladders of mice. C57BL/6J mice were exposed for 1 or 12 weeks to arsenate in drinking water at concentrations of 0.5, 2, 10, and 50 mg As/L. Urinary bladders were analyzed using gene expression microarrays. A consistent reversal was observed in the direction of gene expression change: from predominantly decreased expression at 1 week to predominantly increased expression at 12 weeks. These results are consistent with evidence from in vitro studies of an acute adaptive response that is suppressed on longer exposure due to down-regulation of Fos. Pathways with the highest enrichment in gene expression changes were associated with epithelial-to-mesenchymal transition, inflammation, and proliferation. Benchmark dose (BMD) analysis determined that the lowest median BMD values for pathways were above 5 mg As/L, despite the fact that pathway enrichment was observed at the 0.5 mg As/L exposure concentration. This disparity may result from the non-monotonic nature of the concentration-responses for the expression changes of a number of genes, as evidenced by the much fewer gene expression changes at 2 mg As/L compared to lower or higher concentrations. Pathway categories with concentration-related gene expression changes included cellular morphogenesis, inflammation, apoptosis/survival, cell cycle control, and DNA damage response. The results of this study provide evidence of a concentration-dependent transition in the mode of action for the subchronic effects of Asi in mouse bladder cells in the vicinity of 2 mg Asi/L. Female C57Bl/J mice will be exposed to COLD Arsenate in drinking water. One week interium sac on 7/18/06. 100 samples including liver, lung, kidney and bladder. Bladder will be analyzed with microarrays, 24 samples. Twelve week terminal sac on 10/05/06. 75 total samples including lung, kidney, and bladder. Bladdler will be analyzed by microarray, 25 samples. Drinking water containing As will be prepared weekly with monitoring to determine amount used by mice. Following tissues will be available for genomic study: Bladder, liver, lung, and kidney.

Dataset Information

Identifying Eukaryotes and Factors Influencing Their Biogeography in Drinking Water Metagenomes.

Publications

Identifying Eukaryotes and Factors Influencing Their Biogeography in Drinking Water Metagenomes.

Similar Datasets

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