Biology subjects

Pjevac, P.

Publications and source records attributed to Pjevac, P..

At least 19 recordsLinked to original sources

bioRxiv2026

A Saccharimonadia epibiont interacts with the polyphosphate-accumulating bacterium Candidatus Phosphoribacter in wastewater treatment plants

Many Patescibacteria lineages are abundant in wastewater treatment plants, yet their interaction partners and ecological roles remain poorly understood. Here, we identify a specific association between a novel Saccharimonadia genus, Candidatus Lutisaccharimonas, and the widespread polyphosphate-accumulating organism (PAO) Candidatus Phosphoribacter. Fluorescence in situ hybridization revealed close physical proximity between the two taxa, supporting an epibiotic relationship. Correlated population dynamics, CRISPR spacer matches and evidence of horizontal gene transfer further corroborated this association. Comparative genomic analyses showed that Ca. Lutisaccharimonas has greater metabolic versatility than is typical of Patescibacteria, including complete pathways for de novo nucleotide biosynthesis and genes involved in amino acid metabolism. These metabolic features may shape its interactions with Ca. Phosphoribacter. Together, our findings identify a previously undescribed host association for Patescibacteria, expand current understanding of their metabolic potential, and suggest that the interactions between Saccharimonadia and PAOs may influence phosphorus removal in engineered ecosystems.

microbiology↗

bioRxiv2026

Beyond efficacy: Persistence and off-target effects of three biological nitrification inhibitors in two contrasting agricultural soils

The use of nitrogen (N) fertilizers to meet global food demands is expected to continue rising. However, up to 70% of N applied to agricultural soils is lost through microbially mediated processes such as nitrification. Inhibiting nitrification is thus a key strategy to reduce N losses and improve fertilizer N use efficiency. Various plant-derived compounds, termed biological nitrification inhibitors (BNIs), have been shown to reduce accumulation of nitrification products, intermediates, and byproducts (nitrite, nitrate, nitric and nitrous oxides). However, the mechanisms by which BNIs affect nitrifiers, along with their specificity and persistence in soil are not well understood. Here, we evaluated the effects of three BNIs: methyl 3-(4-hydroxyphenyl) acrylate (MHPA), 6-methoxy-2(3H)-benzoxazolone (MBOA), and limonene, on ammonia-oxidizing, total microbial, and fungal communities in two soils with contrasting pH. Their persistence in each soil was also evaluated. Although ammonia-oxidizing archaea initially dominated nitrifier communities in both soils, their bacterial counterparts significantly increased after mineral N addition but also were more sensitive to BNI application. Limonene and the synthetic inhibitor DMPP stimulated ammonium immobilization, as total soil mineral N was significantly reduced. Limonene and MHPA had the strongest off-target effects, increasing the relative abundance of hydrocarbon-degrading bacteria and potential fungal pathogens, respectively. In contrast, MBOA inhibited nitrification with minimal off-target effects. Among the tested BNIs, MBOA was also the most persistent in the high-pH, high-nitrification-rate soil. Our results show that MBOA is a promising biological inhibitor and highlight the importance of understanding BNIs ecological effects to develop targeted and sustainable N management strategies.

microbiology↗

bioRxiv2026

Potential of exogenous biological nitrification inhibitor addition to improve soil nitrogen availability for crop growth.

Modern agriculture is characterized by substantial fertilizer nitrogen (N) losses from soils, resulting in low crop N-use efficiency. Biological nitrification inhibitors (BNIs) are studied as a strategy to improve N retention in soils by suppressing nitrification. However, the impacts of applying exogenous BNIs to crops with unknown intrinsic BNI capacity remain poorly understood. In this study, we evaluated the impacts of adding three BNIs (methyl 3-(4-hydroxyphenyl) acrylate [MHPA], 6-methoxy-2(3H)-benzoxazolone [MBOA], and limonene), their mixture, and the synthetic nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) on barley (Hordeum vulgare L.) growth, plant and soil N dynamics, and soil microbial communities. Using a rhizobox system with planted and bare-soil compartments, combined with 15N isotope tracing and molecular microbial community analyses, we assessed the spatio-temporal dynamics of N transformations, losses, plant N uptake, and microbial community responses in an alkaline agricultural soil. Independent of inhibitor application, the applied fertilizer N was lost primarily through NO- leaching (3-9% of the applied N). In contrast, N2O emissions represented only 0.001-0.028% of the applied N and varied with inhibitor type. MHPA increased dissolved inorganic N soil pools without affecting plant biomass or 15N uptake or strongly shifting microbial community composition. MBOA reduced NO3- concentrations in soil pore water without influencing plant growth or N uptake but shifted soil microbial community composition. In contrast, limonene reduced plant growth and 15N uptake and most significantly altered microbial community composition, without significantly changing N availability. Applying a BNI mixture, as well as limonene alone, was detrimental to plant growth and 15N uptake. DMPP showed only minor effects on N pools, plant growth, plant N uptake and microbial community composition. Overall, our results reveal both the potential and limitations of exogenous BNI application for improving N retention in crop systems.

microbiology↗

bioRxiv2026

Gut-derived ammonia modulates hypothalamic stress responses during viral infection

The gut-brain axis integrates microbial and host metabolism to regulate systemic physiology, yet its role during viral infection remains poorly defined. Viral infection induces behavioral changes and neuroendocrine stress responses accompanied by profound alterations in gut microbial metabolism. Here, we show that chronic viral infection in mice increases systemic levels of microbiota-derived ammonia in a CD8 T cell-dependent manner. Increased ammonia accumulates in the brain and selectively activates neurons within the paraventricular hypothalamus (PVH), driving corticosterone release into the circulation. Pharmacological inhibition of ammonia detoxification exacerbates these effects, leading to increased corticosterone levels, aggravated sickness behavior, and dampened antiviral responses. Together, these findings identify gut-derived ammonia as a previously unrecognized immunometabolic signal linking antiviral T cell responses to hypothalamic control of systemic stress during viral infection. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=186 SRC="FIGDIR/small/736796v1_ufig1.gif" ALT="Figure 1"> View larger version (59K): org.highwire.dtl.DTLVardef@1291dbcorg.highwire.dtl.DTLVardef@5f223corg.highwire.dtl.DTLVardef@1c0aafaorg.highwire.dtl.DTLVardef@190502_HPS_FORMAT_FIGEXP M_FIG C_FIG

immunology↗

bioRxiv2026

Rapid assessment of nitrification inhibitor efficacy, competitiveness, and specificity using microrespirometry

Nitrification inhibitors are applied to reduce nitrogen losses and greenhouse gas emissions from fertilized agricultural ecosystems. However, their characterization is typically focused on determining effective inhibitor concentrations from growth or substrate conversion assays that are time-intensive and provide limited mechanistic resolution. Here, we present a microrespirometry (MR)-based workflow for rapid mechanistic characterization of nitrification inhibitors using oxygen consumption as a real-time readout for metabolic activity. The workflow enables the simultaneous assessment of inhibitor efficacy, competitiveness, and enzyme specificity within a single experimental setup, as sequential substrate and inhibitor additions enable direct discrimination between competitive and non-competitive inhibition and between ammonia monooxygenase-specific and broader respiratory inhibition. As a proof of concept, we evaluated three known nitrification inhibitors phenylacetylene (PA), nitrapyrin (NP), and dicyandiamide (DCD) using the ammonia-oxidizing bacteria Nitrosomonas europaea and Nitrosospira multiformis, the complete ammonia oxidizer Nitrospira inopinata, and the nitrite oxidizer Nitrospira moscoviensis. We also compared the results from the MR-based inhibition workflow with those from a conventional growth-based approach and observed a poor correlation between results for inhibitors that are not fully enzyme specific. In conclusion, this work establishes MR as a rapid and versatile platform for the mechanistic screening of novel potential nitrification inhibitors. MR assays reproduce known inhibitory responses while substantially reducing experimental time and increasing mechanistic resolution compared to other assays types. Additionally, we provide the first pure-culture characterization of PA, NP, and DCD efficacy and inhibition mechanisms in a complete ammonia oxidizer, N. inopinata.

microbiology↗

bioRxiv2025

Revised 16S rRNA V4 hypervariable region targeting primers enhance detection of Patescibacteria and other lineages across diverse environments

Primer bias in 16S rRNA gene amplicon sequencing can distort microbial diversity estimates by underrepresenting key taxa. We introduce a modified primer pair (V4-EXT) targeting the hypervariable V4 region of bacterial and archaeal 16S rRNA genes, with improved in silico taxonomic inclusivity. To benchmark performance, we analyzed 938 samples from terrestrial, aquatic, and host-associated habitats, comparing microbial community profiles derived with V4-EXT and the currently most widely used V4-targeted primers. V4-EXT substantially improved the detection of Patescibacteria and other underrepresented lineages, such as Chloroflexota and Iainarchaeota, while enhancing recovery of novel amplicon sequence variants across sample types. Overall, V4-EXT provides broader taxonomic coverage and more inclusive microbial community profiles, particularly in high-diversity ecosystems such as groundwater and soils. We propose V4-EXT as a robust successor for comprehensive microbial community analysis across diverse habitats.

microbiology↗

bioRxiv2025

Branched-chain amino acid assimilation promotes mixotrophy of ammonia-oxidizing archaeal sponge symbionts

Ammonia-oxidizing archaea (AOA) frequently form symbiotic associations with marine sponges. While free-living AOA are generally considered metabolically constrained chemolithoautotrophs, sponge-associated AOA encode for a branched-chain amino acid (BCAA) transporter, suggesting mixotrophic potential. Here, we test the unusual mixotrophic lifestyle of sponge-associated AOA by tracing the assimilation of 13C- and 15N-labeled BCAA in the sponge holobiont Ianthella basta. We demonstrate that BCAA degradation fuels ammonia oxidation and quantify BCAA uptake at the single-cell level by combining stable isotope probing, catalyzed reporter deposition fluorescence in situ hybridization, and nanoscale secondary ion mass spectrometry. Our results reveal that sponge-associated AOA are mixotrophic, assimilating BCAA as an additional carbon and nitrogen source. This metabolic adaptation may modulate BCAA availability in the holobiont, potentially regulating the hosts mTOR pathway. Collectively, our study reveals a novel nutritional interaction in sponge holobionts and challenges the perception of constrained metabolic capacities of AOA.

microbiology↗

bioRxiv2025

Dynamic changes of protist community composition along a surface water-groundwater transect in the Danube wetland Lobau, Vienna, Austria

Groundwater remains an unexplored habitat in terms of protistan diversity and community dynamics, when compared to better-studied surface aquatic environments. To address this knowledge gap, a comparison of protistan communities found at two differing sites, a wetland surface water body and the close-by shallow aquifer, aims to shed light on environmental drivers of both the total and active protist communities using molecular analyses. The two-year time series with monthly samplings provided insight into seasonal pattern, and barcoding allowed taxonomic affiliation. Our study revealed clear differences in community composition and trophic modes with the habitat type. Protistan communities in shallow groundwater exhibited pronounced seasonal dynamics somehow temporally linked to the surface water counterparts. Higher absolute water levels in the backwater compared to groundwater and the significant fraction of phototrophic protists sampled from the shallow aquifer point at groundwater recharge from surface water to strongly influence protistan community composition.

microbiology↗

bioRxiv2025

Associations between gut microbiota and personality traits: insights from a captive common marmoset (Callithrix jacchus) colony

Recent studies have suggested the link between inter-individual behavioural variation (i.e., animal personality) and gut microbiota. Non-human primates living under controlled conditions are valuable models to investigate diet-independent microbiome-host interactions. In this study, we investigated links between specific gut microbiota members and personality traits, as well as group membership, sex, age class, breeding status and relatedness of 26 captive common marmosets (Callithrix jacchus), maintained under the same diet and housing conditions. Personality was assessed using an established testing battery in repeated tests. Then, we collected a total of 225 fecal samples during the summers of 2017 and 2019 from five marmoset social groups for 16S rRNA gene amplicon sequencing. Within-individual microbiota variance was smaller than that between group members. Group members also exhibited more similar gut microbiota than individuals from different groups in each sampling year. Beta diversity of the gut microbiota was linked with personality traits, age class, sex, and breeding status, but not with genetic relatedness. We identified specific bacterial taxa associated with personality traits. In particular, members of the sulfite-reducing genera Desulfovibrio were enriched in more avoidant marmosets. Amplicon sequencing of the dissimilatory sulfite reductase gene dsrB confirmed this pattern, yet additionally revealed an unknown uncultured bacterium that was the predominant sulfite-reducing bacterium in the fecal samples and was linked to more explorative individuals. These findings highlight specific association patterns between selected microbial taxa and personality traits in captive common marmosets. ImportanceThis study provides valuable insights into the intricate relationship between gut microbiota and host personality traits, using captive common marmosets as a model. By controlling for diet and housing conditions, it probes key host factors such as personality, age, sex, and social group membership, offering a robust framework for understanding microbiome-host interactions. The discovery of specific microbial taxa associated with personality traits, particularly the enrichment of sulfite-reducing genera in more avoidant individuals, underscores the potential role of the gut microbiome in shaping or reflecting personality. These findings advance our understanding of microbiome-host dynamics and pave the way for future research on the mechanistic links between behavior and gut microbiota in other animal models and across broader ecological contexts.

microbiology↗

bioRxiv2024

Human-derived microRNA 21 regulates indole and L-tryptophan biosynthesis transcripts in a prominent gut symbiont

In the gut, microRNAs (miRNAs) produced by intestinal epithelial cells are secreted into the lumen and can shape the composition and function of the gut microbiome. Crosstalk between gut microbes and the host plays a key role in irritable bowel syndrome (IBS) and inflammatory bowel diseases, yet little is known about how the miRNA-gut microbiome axis contributes to the pathogenesis of these conditions. In this study, we aimed to explore the ability of miR-21, a miRNA that we found decreased in stool samples from IBS patients, to associate with and regulate gut microbiome function. Incubation of human faecal microbiota with miR-21 revealed a rapid association with microbial cells, reproducible across multiple donor samples. Fluorescence-activated cell sorting and sequencing of microbial cells incubated with fluorescently-labelled miR-21 identified organisms belonging to the genera Bacteroides, Limosilactobacillus, Ruminococcus, or Coprococcus which predominantly interacted with miR-21. Surprisingly, these and other genera also interacted with a miRNA scramble control, suggesting that physical interaction and/or uptake of these miRNAs by gut microbiota is not sequence-dependent. Nevertheless, transcriptomic analysis of the gut commensal Bacteroides thetaiotaomicron revealed a miRNA sequence-specific effect on bacterial transcript levels. Supplementation of miR-21, but not of small RNA controls resulted in significantly altered levels of many cellular transcripts and increased transcription of a biosynthetic operon for indole and L-tryptophan, metabolites known to regulate host inflammation and colonic motility. Our study identifies a novel putative miR-21-dependent pathway of regulation of intestinal function through the gut microbiome with implications for gastrointestinal conditions.

microbiology↗

bioRxiv2024

Niche and spatial partitioning restrain ecological equivalence among microbes along aquatic redox gradient

Microbial metabolic capabilities and interactions shape their niche hypervolume that in turn governs their ecological strategies and ecosystem services. In the context of functional redundancy or ecological equivalence, the focus has been on functional guilds in order to bypass the complex challenge faced by niche theory for disentangling the niche hypervolume. However, in some cases this simplification has been at the expense of ignoring the role of individual genotype of each microbe within a functional guild and fails to explain how the diversity within each functional guild is maintained. In this study, we inspect the metabolic profile of metagenome-assembled genomes along the pronounced redox gradient of the water column in an anchialine cave. Bridging neutral theory of biodiversity and biogeography and niche theory, our analysis uses focal metabolic capabilities while also incorporating individuality by looking into background metabolic capabilities of each individual and further includes spatial distribution of microbes to delineate their niche space. Our results emphasize that differences in background metabolic capabilities are critical for furnishing the niche hypervolume of microbes carrying the same focal metabolic capability and refute their ecological equivalence with their spatial distribution further enables niche partitioning among them.

ecology↗

bioRxiv2024

Distinct microbial communities are linked to organic matter properties in millimetre-sized soil aggregates

Soils provide essential ecosystem services and represent the most diverse habitat on Earth. It has been suggested that the presence of various physico-chemically heterogenous microhabitats supports the enormous diversity of microbial communities in soil. However, little is known about the relationship between microbial communities and their immediate environment at the micro- to millimetre-scale. In this study, we examined whether bacteria, archaea, and fungi organise into distinct communities in individual 2-millimetre-sized soil aggregates and compared them to communities of homogenized bulk soil samples. Furthermore, we investigated their relationship to their local environment by concomitantly determining microbial community structure and physico-chemical properties from the same individual aggregates. Aggregate-communities displayed exceptionally high beta-diversity, with 3-4 aggregates collectively capturing more diversity than their homogenized parent soil core. Up to 20-30% of ASVs (particularly rare ones) were unique to individual aggregates selected within a few centimetres. Aggregates and bulk soil samples showed partly different dominant phyla, indicating that taxa that are potentially driving biogeochemical processes at the small scale may not be recognized when analysing larger soil volumes. Microbial community composition and richness of individual aggregates were closely related to aggregate-specific carbon and nitrogen content, carbon stable-isotope composition, and soil moisture, indicating that aggregates provide a stable environment for sufficient time to allow co-development of communities and their environment. We conclude that the soil microbiome is a metacommunity of variable subcommunities. Our study highlights the necessity to study small, spatially coherent soil samples to better understand controls of community structure and community-mediated processes in soils.

ecology↗

bioRxiv2023

Novel nitrifying symbiont lineages are vertically inherited and widespread in marine sponges

Ammonia-oxidising archaea and nitrite-oxidising bacteria are common members of marine sponge microbiomes. They derive energy for carbon fixation and growth from nitrification - the oxidation of ammonia to nitrite and further to nitrate - and are proposed to play essential roles in the carbon and nitrogen cycling of sponge holobionts. In this study, we characterise two novel nitrifying symbiont lineages, Ca. Nitrosokoinonia and Ca. Nitrosymbion in the marine sponge Coscinoderma matthewsi using a combination of molecular tools, single-cell imaging techniques, and physiological rate measurements. Both represent a new genus in the ammonia-oxidising archaeal class Nitrososphaeria and the nitrite-oxidising bacterial order Nitrospirales, respectively. Furthermore, we show that larvae of this viviparous sponge are densely colonised by representatives of Ca. Nitrosokoinonia and Ca. Nitrosymbion indicating vertical transmission. In adults, the representatives of both symbiont genera are located extracellularly in the mesohyl. Comparative metagenome analyses and physiological data suggest that ammonia-oxidising archaeal symbionts of the genus Ca. Nitrosokoinonia strongly rely on endogenously produced nitrogenous compounds (i.e., ammonium, urea, nitriles/cyanides, and creatinine) rather than on exogenous ammonium sources taken up by the sponge. Additionally, the nitrite-oxidising bacterial symbionts Ca. Nitrosymbion may reciprocally support the ammonia-oxidisers with ammonia via the utilisation of sponge-derived urea and cyanate. Interestingly, comparative analyses of published environmental 16S rRNA amplicon data revealed that Ca. Nitrosokoinonia and Ca. Nitrosymbion are widely distributed and predominantly associated with marine sponges and corals, suggesting a broad relevance of our findings.

microbiology↗

bioRxiv2023

Global abundance patterns, diversity, and ecology of Patescibacteria in wastewater treatment plants

BackgroundMicroorganisms are responsible for nutrient removal and resource recovery in wastewater treatment plants (WWTPs), and their diversity is often studied by 16S rRNA gene amplicon sequencing. However, this approach underestimates the abundance and diversity of Patescibacteria due to the low coverage of commonly used PCR primers for this highly divergent bacterial phylum. Therefore, our current understanding of the global diversity, distribution, and ecological role of Patescibacteria in WWTPs is very incomplete. This is particularly relevant as Patescibacteria are considered to be associated with microbial host cells and can therefore influence the abundance and temporal variability of other microbial groups that are important for WWTP functioning. ResultsHere, we evaluated the in silico coverage of widely used 16S rRNA gene-targeted primer pairs and redesigned a primer pair targeting the V4 region of bacterial and archaeal 16S rRNA genes to expand its coverage for Patescibacteria. We then experimentally evaluated and compared the performance of the original and modified V4-targeted primers on 565 WWTP samples from the MiDAS global sample collection. Using the modified primer pair, the percentage of ASVs classified as Patescibacteria increased from 5.9% to 23.8%, and the number of detected patescibacterial genera increased from 560 to 1,576, while the detected diversity of the remaining microbial community remained similar. Due to this significantly improved coverage of Patescibacteria, we identified 23 core genera of Patescibacteria in WWTPs and described the global distribution pattern of these unusual microbes in these systems. Finally, correlation network analysis revealed potential host organisms that might be associated with Patescibacteria in WWTPs. Interestingly, strong indications were found for an association between Patescibacteria of the Saccharimonadia and globally abundant polyphosphate-accumulating organisms of the genus Ca. Phosporibacter. ConclusionsOur study (i) provides an improved 16S rRNA gene V4 region-targeted amplicon primer pair inclusive of Patescibacteria with little impact on the detection of other taxa, (ii) reveals the diversity and distribution patterns of Patescibacteria in WWTPs on a global scale, and (iii) provides new insights into the ecological role and potential hosts of Patescibacteria in WWTPs.

ecology↗

bioRxiv2022

A novel taurine-respiring murine gut bacterium contributes to colonization resistance against enteropathogens

Taurine-respiring gut bacteria produce H2S with ambivalent impact on host health. We report the isolation and genomic-ecophysiological characterization of the first taurine-respiring mouse gut bacterium. Taurinivorans muris represents a new widespread species with protective capacity against pathogens and differs from the human gut sulfidogen Bilophila wadsworthia in its sulfur metabolism and host distribution. Despite alternative physiologies, taurine respiration was the main in vivo lifestyle of T. muris independent of mouse diet and genotype. In gnotobiotic mice, T. muris selectively enhanced the activity of a sulfur metabolism gene-encoding prophage and provided slightly increased colonization resistance against Salmonella Typhimurium, which showed reduced expression of galactonate catabolism genes. We identified T. muris as the dominant sulfidogen of a mouse microbiota that conferred H2S-mediated protection against Klebsiella pneumoniae in a previous study. Together, we revealed the realized physiological niche of a key murine gut sulfidogen and its impact on pathogen and phage gene expression. One sentence summaryOur work identified and characterized a new core member of the murine gut microbiota, revealed sulfidogenic taurine respiration as its predominant in vivo lifestyle, and emphasizes its protective function in pathogen colonization.

microbiology↗

bioRxiv2021

Ammonia-oxidizing archaea possess a wide range of cellular ammonia affinities

Nitrification, the oxidation of ammonia to nitrate, is an essential process in the biogeochemical nitrogen cycle. The first step of nitrification, ammonia oxidation, is performed by three, often co- occurring guilds of chemolithoautotrophs: ammonia-oxidizing bacteria (AOB), archaea (AOA), and complete ammonia oxidizers (comammox). Substrate kinetics are considered to be a major niche-differentiating factor between these guilds, but few AOA strains have been kinetically characterized. Here, the ammonia oxidation kinetic properties of 12 AOA representing all major phylogenetic lineages were determined using microrespirometry. Members of the genus Nitrosocosmicus have the lowest substrate affinity of any characterized AOA, which are similar to previously determined affinities of AOB. This contrasts previous assumptions that all AOA possess much higher substrate affinities than their comammox or AOB counterparts. The substrate affinity of ammonia oxidizers correlated with their cell surface area to volume ratios. In addition, kinetic measurements across a range of pH values strongly supports the hypothesis that - like for AOB - ammonia and not ammonium is the substrate for the ammonia monooxygenase enzyme of AOA and comammox. Together, these data will facilitate predictions and interpretation of ammonia oxidizer community structures and provide a robust basis for establishing testable hypotheses on competition between AOB, AOA, and comammox.

microbiology↗

bioRxiv2020

A refined set of rRNA-targeted oligonucleotide probes for in situ detection and quantification of ammonia-oxidizing bacteria

Ammonia-oxidizing bacteria (AOB) of the betaproteobacterial genera Nitrosomonas and Nitrosospira are key nitrifying microorganisms in many natural and engineered ecosystems. Since many AOB remain uncultured, fluorescence in situ hybridization (FISH) with rRNA-targeted oligonucleotide probes has been one of the most widely used approaches to study the community composition, abundance, and other features of AOB directly in environmental samples. However, the established and widely used AOB-specific 16S rRNA-targeted FISH probes were designed up to two decades ago, based on much smaller rRNA gene sequence datasets than available today. Several of these probes cover their target AOB lineages incompletely and suffer from a weak target specificity, which causes cross-hybridization of probes that should detect different AOB lineages. Here, a set of new highly specific 16S rRNA-targeted oligonucleotide probes was developed and experimentally evaluated that complements the existing probes and enables the specific detection and differentiation of the known, major phylogenetic clusters of betaproteobacterial AOB. The new probes were successfully applied to visualize and quantify AOB in activated sludge and biofilm samples from seven pilot- and full-scale wastewater treatment systems. Based on its improved target group coverage and specificity, the refined probe set will facilitate future in situ analyses of AOB.

microbiology↗

bioRxiv2019

Transcriptomic response of Nitrosomonas europaea transitioned from ammonia- to oxygen-limited steady-state growth

Ammonia-oxidizing microorganisms perform the first step of nitrification, the oxidation of ammonia to nitrite. The bacterium Nitrosomonas europaea is the best characterized ammonia oxidizer to date. Exposure to hypoxic conditions has a profound effect on the physiology of N. europaea, e.g. by inducing nitrifier denitrification, resulting in increased nitric and nitrous oxide production. This metabolic shift is of major significance in agricultural soils, as it contributes to fertilizer loss and global climate change. Previous studies investigating the effect of oxygen limitation on N. europaea have focused on the transcriptional regulation of genes involved in nitrification and nitrifier denitrification. Here, we combine steady-state cultivation with whole genome transcriptomics to investigate the overall effect of oxygen limitation on N. europaea. Under oxygen-limited conditions, growth yield was reduced and ammonia to nitrite conversion was not stoichiometric, suggesting the production of nitrogenous gases. However, the transcription of the principal nitric oxide reductase (cNOR) did not change significantly during oxygen-limited growth, while the transcription of the nitrite reductase-encoding gene (nirK) was significantly lower. In contrast, both heme-copper containing cytochrome c oxidases encoded by N. europaea were upregulated during oxygen-limited growth. Particularly striking was the significant increase in transcription of the B-type heme-copper oxidase, proposed to function as a nitric oxide reductase (sNOR) in ammonia-oxidizing bacteria. In the context of previous physiological studies, as well as the evolutionary placement of N. europaeas sNOR with regards to other heme-copper oxidases, these results suggest sNOR may function as a high-affinity terminal oxidase in N. europaea and other AOB.\n\nImportanceNitrification is a ubiquitous, microbially mediated process in the environment and an essential process in engineered systems such as wastewater and drinking water treatment plants. However, nitrification also contributes to fertilizer loss from agricultural environments increasing the eutrophication of downstream aquatic ecosystems and produces the greenhouse gas nitrous oxide. As ammonia-oxidizing bacteria are the most dominant ammonia-oxidizing microbes in fertilized agricultural soils, understanding their response to a variety of environmental conditions is essential for curbing the negative environmental effects of nitrification. Notably, oxygen limitation has been reported to significantly increase nitric oxide and nitrous oxide production during nitrification. Here we investigate the physiology of the best characterized ammonia-oxidizing bacterium, Nitrosomonas europaea, growing under oxygen-limited conditions.

microbiology↗