Project description:Anaerobic ammonium oxidation (anammox) emerges as a sustainable solution for nitrogen removal in sewage, but it is susceptible to stress induced by xenobiotics that are ubiquitous in sewage. Despite wide recognition of this critical issue, a comprehensive understanding of the molecular and ecological mechanisms underlying the response of anammox consortia to xenobiotic stress remain elusive. Here, we integrated multi-omics approaches with biofilm reactor operation to unravel how bisphenol A (BPA, a representative xenobiotic) perturbs anammox consortia across environmentally relevant concentrations. We show that anammox consortia tolerated low BPA levels (0.2–2 mg/L), where nitrogen removal efficiency transiently declined and subsequently recovered, aided by a 30.9% increase in quorum-sensing (QS) signal C6-HSL. By contrast, exposure to ≥10 mg/L BPA caused severe and irreversible inhibition, with total inorganic nitrogen removal dropping to 17.8%. High BPA concentrations suppressed QS signaling, intensified oxidative stress, and compromised membrane integrity, leading to enzymatic inhibition and transcriptional repression of anammox functional genes. Multi-omics evidence revealed that BPA stress also promoted horizontal transfer of the BPA-degrading gene bisdA via extracellular DNA, suggesting a new community-level adaptive mechanism. Metagenomic and metabolomic analyses further indicated BPA-driven restructuring of microbial networks, namely high BPA levels favored denitrifiers and BPA degraders while suppressing anammox bacteria, and triggered metabolic reprogramming toward xenobiotic degradation at the expense of nucleotide and amino acid biosynthesis. Together, these findings reveal a multifaceted collapse mechanism of anammox under BPA stress, providing a mechanistic basis for designing strategies to safeguard microbial nitrogen removal in xenobiotic-laden wastewaters.
Project description:Anaerobic ammonium-oxidising (anammox) bacteria, members of the ‘Candidatus Brocadiaceae’ family, play an important role in the nitrogen cycle and are estimated to be responsible for about half of the oceanic nitrogen loss to the atmosphere. Anammox bacteria combine ammonium with nitrite and produce dinitrogen gas via the intermediates nitric oxide and hydrazine (anammox reaction) while nitrate is formed as a by-product. These reactions take place in a specialized, membrane-bound compartment called the anammoxosome. Therefore, the substrates ammonium, nitrite and product nitrate have to cross the outer-, cytoplasmic- and anammoxosome membranes to enter or exit the anammoxosome. The genomes of all anammox species harbour multiple copies of ammonium-, nitrite- and nitrate transporter genes. Here we investigated how the distinct genes for ammonium-, nitrite- and nitrate- transport were expressed during substrate limitation in membrane bioreactors. Transcriptome analysis of Kuenenia stuttgartiensis planktonic cells under ammonium-limitation showed that three of the seven ammonium transporter genes and one of the six nitrite transporter genes were significantly upregulated, while another ammonium and nitrite transporter gene were downregulated in nitrite limited growth conditions. The two nitrate transporters were expressed to similar levels in both conditions. In addition, genes encoding enzymes involved in the anammox reaction were differentially expressed, with those using nitrite as a substrate being upregulated under nitrite limited growth and those using ammonium as a substrate being upregulated during ammonium limitation. Taken together, these results give a first insight in the potential role of the multiple nutrient transporters in regulating transport of substrates and products in and out of the compartmentalized anammox cell.
Project description:The community composition (in terms of abundance, distribution and contribution of diverse clades) of bacteria involved in nitrogen transformations in the oxygen minimum zones may be related to the rates of fixed N loss in these systems. The abundance of both denirifying and anammox bacteria, and the assemblage composition of denitrifying bacteria were investigated in the Eastern Tropical South Pacific and the Arabian Sea using assays based on molecular markers for the two groups of bacteria. The abundance and distribution of bacteria associated with the fixed N removal processes denitrification and anammox were investigated using quantitative PCR for genes encoding nitrite reductase (nirK and nirS) in denitrifying bacteria and hydrazine oxidase(hzo) and 16S rRNA genesin anammox bacteria. All of these genes had depth distributions with maxima associated with the secondary nitrite maximum in low oxygen waters. NirS was mch more abundant than nirK, and much more abundant than the 16S rRNA gene from anammox bacteria. The ratio of hzo:16S rRNA for anammox was low and variable implying greater unexplored diversity in the the hzo gene. Assemblage composition of the abundant nirS-type denitrifiers was evaluated using a funcitonal gene microarray. Of the nirS archetypes represented on the microarray, very few occurred speficically in one region or depth interval, but the assemblages varied significantly. Community composition of denitrifiers based on microarray analysis of the nirS gene was most different between geographical regions. Within each region, the surface layer and OMZ assemblages clustered distinctly. Thus, in addition to spatial and temporal variation in denitrificaiton and anammox rates, both microbial abundance and community composition also vary between OMZ regions and depths.
Project description:Four Fe(II) concentrations (0.03, 0.09, 0.12 & 0.75 mM) were tested to investigate the stimulation and inhibition effects of ferrous iron on anammox bacterial activity. RNAs were extracted from the cultures, and the synthesized cDNAs by reverse transcription were used to carry out GeoChip analysis, by which the functional communities and expression level differences in functional genes under different Fe(II) concentrations conditions were obtained, and the response of anammox bacteria to Fe(II) stimulation and inhibition are speculated.