Ladderane lipids synthesised by anammox bacteria hold significant potential for applications in jet fuel, drug delivery, and optoelectronics. Despite the widespread use of anammox bacteria in nitrogen removal from wastewater, the optimal conditions for maximising ladderane production remain unclear, limiting their broader application. To address this, we operated a fed-batch bioreactor with anammox bacteria, gradually adjusting the pH from 6.5 to 7.5 while regularly sampling for microbial community composition (Illumina sequencing), proteins, and ladderane lipids (UHPLC-HRMS). Our findings reveal that ladderane production positively correlates with rising pH increasing nearly fivefold as pH rose from 6.5 to 7.5, with a notable shift towards lipids containing two ladderane alkyl chains at higher pH. However, the conditions at an alkaline pH range also induced mild stress in anammox bacteria, as evidenced by our proteomic and microbial community data. Therefore, we propose maintaining a pH above 7.5 to enrich ladderane-rich anammox biomass but emphasise the need for gradual adaptation. This approach could optimise anammox installations for producing high-value ladderane lipids from wastewater.
- Klíčová slova
- Alkalinity, Anammox bacteria, Biorefinery, Ladderane lipids, Wastewater treatment, pH,
- MeSH
- Bacteria * metabolismus MeSH
- biomasa MeSH
- bioreaktory * MeSH
- koncentrace vodíkových iontů MeSH
- lipidy * chemie MeSH
- odpadní voda * chemie MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- lipidy * MeSH
- odpadní voda * MeSH
Traditional wastewater treatment often fails to remove pharmaceuticals, necessitating advanced solutions, such as TiO2 photocatalysis, for post-treatment. However, conventionally applied powder TiO2 can be cumbersome to separate from treated water. To solve this issue, this study immobilized three TiO2 photocatalysts (Anatase 16, Anatase 5, and P25) into porous layers and evaluated their efficacy for the degradation of three pharmaceuticals (naproxen, NPX; sulfamethoxazole, SMX; metformin, MTF) in standard solutions and greywater pretreated in a membrane bioreactor (MBR). In standard solutions, photocatalysis tests revealed a high degradation efficacy (NPX 100%, SMX 76-95%, MTF 57-75%) and challenged the belief that OH• is the predominant reactive oxygen species (ROS). The primary ROS were 1O2 for NPX and OH• for SMX and MTF. The raw greywater (NPX, SMX, MTF - 0.5 mg·L-1) treatment in MBR removed only 17-22% of the pharmaceuticals, highlighting the need for post-treatment. Using this pretreated greywater, P25 layers excelled for NPX (78 ± 5%) and SMX (73 ± 4%) but were less effective for MTF (40 ± 16%) compared to Anatase 16 (60 ± 10%). Moreover, the effluent toxicity (Aliivibrio fischeri) was reduced, and the degradation products were identified. Overall, TiO2 layers are a high-potential method for removing pharmaceuticals from MBR-treated greywater.
- Publikační typ
- časopisecké články MeSH
The early detection of upcoming disease outbreaks is essential to avoid both health and economic damage. The last four years of COVID-19 pandemic have proven wastewater-based epidemiology is a reliable system for monitoring the spread of SARS-CoV-2, a causative agent of COVID-19, in an urban population. As this monitoring enables the identification of the prevalence of spreading variants of SARS-CoV-2, it could provide a critical tool in the fight against this viral disease. In this study, we evaluated the presence of variants and subvariants of SARS-CoV-2 in Prague wastewater using nanopore-based sequencing. During August 2021, the data clearly showed that the number of identified SARS-CoV-2 RNA copies increased in the wastewater earlier than in clinical samples indicating the upcoming wave of the Delta variant. New SARS-CoV-2 variants consistently prevailed in wastewater samples around a month after they already prevailed in clinical samples. We also analyzed wastewater samples from smaller sub-sewersheds of Prague and detected significant differences in SARS-CoV-2 lineage progression dynamics among individual localities studied, e.g., suggesting faster prevalence of new variants among the sites with highest population density and mobility.
- Klíčová slova
- Epidemiology, Nanopore-based sequencing, Prague, SARS-CoV-2, Variants, Wastewater,
- MeSH
- COVID-19 * epidemiologie MeSH
- lidé MeSH
- nanopóry * MeSH
- odpadní voda MeSH
- pandemie MeSH
- prevalence MeSH
- RNA virová MeSH
- SARS-CoV-2 genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- odpadní voda MeSH
- RNA virová MeSH
This review critically assesses nitrogen removal technologies applied in the reject water treatment, across different stages of technological development, with a focus on their economic and environmental impacts. The prevalent use of biological processes raises concerns due to potential environmental impacts caused by N2O emissions. However, partial nitritation-anaerobic ammonium oxidation demonstrated economic benefits and the potential for positive environmental outcomes when properly operated and controlled. Furthermore, reject water, in many cases, provides sufficient nitrogen concentrations for nitrogen recovery processes, such as ammonia stripping, substituting production of industrial fertilizers and contributing to a circular economy. Nonetheless, their financial competitiveness is subject to various conditions, including the nitrogen concentration or reject water flow. As the environmental benefits of bioprocesses and economic benefits of nitrogen recovery processes may vary, it is crucial to further optimize both and investigate novel promising technologies such as electrochemical systems, denitrifying anaerobic methane oxidation or direct ammonia oxidation.
- Klíčová slova
- Ammonium nitrogen, Life-cycle assessment, Nitrous oxide, Sidestream treatment, Technology readiness,
- MeSH
- amoniak * MeSH
- bioreaktory MeSH
- denitrifikace * MeSH
- dusík MeSH
- oxidace-redukce MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- amoniak * MeSH
- dusík MeSH
Monkeypox virus (Mpxv) is a dsDNA virus that has become a global concern for human health in 2022. As both infected people and non-human hosts can shed the virus from their skin, faeces, urine and other body fluids, and the resulting sewage contains viral load representative of the whole population, it is highly promising to detect the spread of monkeypox virus in municipal wastewater. We established a methodology for sewage-based monitoring of Mpxv in Prague and analysed samples (n = 24) already early August-October of 2022 in a municipality with 1.4 million inhabitants that only reported 29 cumulative cases in this period. We isolated Mpxv DNA with the Wizard Enviro Total Nucleic Acid Kit, and thereafter detected Mpxv DNA using the EliGene® Monkeypox RT-PCR Kit. Prague wastewater was positive for Mpxv (in total 9 positive samples in periods with 1-9 new cases per week, coinciding with a weekly incidence of 0.07-0.64 per 100,000 inhabitants. The method for confirmation of wastewater positivity via semi-nested PCR and Sanger sequencing was successfully confirmed on positive controls including Mpxv particles and Mpxv-positive wastewater from the Netherlands. However, for Prague wastewater samples, amplification of Mpxv DNA via semi-semi-nested PCR was unsuccessful. This was probably due to extremely low case count, leading to the amplification of non-target bacterial DNA. Compared to other studies with much higher Mpxv prevalence, we show the outstanding sensitivity of our approach for monitoring the spread of monkeypox using wastewater.
- Klíčová slova
- Monkeypox, Mpox, Sewage, Virus, Wastewater-based epidemiology,
- MeSH
- DNA virů genetika MeSH
- lidé MeSH
- odpadní voda MeSH
- odpadní vody MeSH
- opičí neštovice * diagnóza MeSH
- virus opičích neštovic genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- DNA virů MeSH
- odpadní voda MeSH
- odpadní vody MeSH
To protect the environment and human health, antibiotic resistance genes (ARGs) and persistent pharmaceuticals need to be removed from WWTP effluent prior to its reuse. However, an efficient process for removing free-floating extracellular DNA (exDNA) in combination with a wide range of pharmaceuticals is yet to be reported for real process conditions. As a possible solution, we treated real ultrafiltered WWTP effluent with UV/H2O2 and combined GAC and zeolite sorption. In terms of exDNA, sequencing and high-throughput quantitative PCR (HT-qPCR) showed that exDNA is a potent carrier of numerous ARGs in ultrafiltered WWTP effluent (123 ARGs), including multi-drug efflux pump mexF that became the dominant exARG in GAC effluent over time. Due to the exposure to degradation agents, exDNA was reduced more efficiently than intracellular DNA, and overall levels of ARGs were substantially lowered. Moreover, GAC sorption was particularly effective in the removal of almost all the 85 detected pharmaceutical residues, with fresh GAC demonstrating an efficiency of up to 100%. However, zeolite (Si/Al 0.8) addition was needed to enhance the removal of persistent pollutants such as gabapentin and diclofenac to 57% and up to 100%, respectively. Our combined approach eminently decreases the hazardous effects of pharmaceuticals and antibiotic resistance in the ultrafiltered WWTP effluent, producing effluent suitable for multiple reuse options according to the latest legislation. In addition, we provided similarly promising but less extensive data for surface water and treated greywater.
- Klíčová slova
- Advanced oxidation process, Extracellular antimicrobial resistance genes, Micropollutants, Municipal effluent, Water reuse, Zeolite,
- MeSH
- antibakteriální látky * farmakologie MeSH
- antibiotická rezistence genetika MeSH
- bakteriální geny MeSH
- léčivé přípravky MeSH
- lidé MeSH
- odpadní voda MeSH
- peroxid vodíku chemie MeSH
- zeolity * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antibakteriální látky * MeSH
- léčivé přípravky MeSH
- odpadní voda MeSH
- peroxid vodíku MeSH
- zeolity * MeSH
The adaptation of bacteria involved in anaerobic ammonium oxidation (anammox) to low temperatures will enable more efficient removal of nitrogen from sewage across seasons. At lower temperatures, bacteria typically tune the synthesis of their membrane lipids to promote membrane fluidity. However, such adaptation of anammox bacteria lipids, including unique ladderane phospholipids and especially shorter ladderanes with absent phosphatidyl headgroup, is yet to be described in detail. We investigated the membrane lipids composition (UPLC-HRMS/MS) and dominant anammox populations (16S rRNA gene amplicon sequencing, Fluorescence in situ hybridization) in 14 anammox enrichments cultivated at 10-37 °C. "Candidatus Brocadia" appeared to be the dominant organism in all but two laboratory enrichments of "Ca. Scalindua" and "Ca. Kuenenia". At lower temperatures, the membranes of all anammox populations were composed of shorter [5]-ladderane ester (reduced chain length demonstrated by decreased fraction of C20/(C18 + C20)). This confirmed the previous preliminary evidence on the prominent role of this ladderane fatty acid in low-temperature adaptation. "Ca. Scalindua" and "Ca. Kuenenia" had distinct profile of ladderane lipids compared to "Ca. Brocadia" biomasses with potential implications for adaptability to low temperatures. "Ca. Brocadia" membranes contained a much lower amount of C18 [5]-ladderane esters than reported in the literature for "Ca. Scalindua" at similar temperature and measured here, suggesting that this could be one of the reasons for the dominance of "Ca. Scalindua" in cold marine environments. Furthermore, we propose additional and yet unreported mechanisms for low-temperature adaptation of anammox bacteria, one of which involves ladderanes with absent phosphatidyl headgroup. In sum, we deepen the understanding of cold anammox physiology by providing for the first time a consistent comparison of anammox-based communities across multiple environments.
- Klíčová slova
- Anaerobic ammonium oxidation, Candidatus Brocadia, Candidatus Scalindua, Effect of temperature, Ladderane phospholipids,
- MeSH
- anaerobióza MeSH
- anaerobní oxidace amoniaku * MeSH
- Bacteria * MeSH
- hybridizace in situ fluorescenční MeSH
- membránové lipidy MeSH
- oxidace-redukce MeSH
- RNA ribozomální 16S genetika MeSH
- teplota MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- membránové lipidy MeSH
- RNA ribozomální 16S MeSH
Partial nitritation-anammox (PN/A) process will substantially reduce the costs for the removal of nitrogen in the mainstream of municipal sewage. However, one of the mainstream PN/A challenges is to reduce the time necessary for the adaptation of anammox bacteria to lower temperatures in mild climates. In this study, we exposed anammox flocculent culture to cold shocks [35°C → 5°C (8 h) → 15°C] and evaluated long-term cold shock response. Over a post-shock period of 40 d at 15°C, the nitrogen removal rates in the shocked culture were significantly higher compared to control, with maximum rates up to 0.082 and 0.033 kg-N/kg-VSS/d or 0.164 and 0.076 kg-N/m3/d, for shocked culture and control, respectively. In the corresponding semi-batch cycles, the shocked culture was on average 136 ± 101% more active than the control, due to the negative effect of cold shock on side populations and more active anammox cells. Per FISH, Ca. Brocadia anammoxidans and Ca. Scalindua survived the shock and remained present throughout. Thus, both anammox microorganisms seem to respond favourably to cold shocks. In sum, we provide further evidence that cold shocks accelerate the adaptation of anammox to the mainstream of municipal WWTPs. Further, for the first time, we report the long-term adaptive response of anammox to cold shocks.
- Klíčová slova
- Anammox, adaptation, cold shock response, low temperature, mainstream,
- Publikační typ
- časopisecké články MeSH
Partial nitritation/anammox can provide energy-efficient nitrogen removal from the main stream of municipal wastewater. The main bottleneck is the growth of nitrite oxidizing bacteria (NOB) at low temperatures (<15 °C). To produce effluent suitable for anammox, real municipal wastewater after anaerobic pretreatment was treated by enriched ammonium oxidizing bacteria (AOB) in suspended sludge SBR at 12 °C. NOB were continually washed out using aerobic duration control strategy (ADCS). Solids retention time was set to 9-16 days. Using this approach, average ammonia conversion higher than 57% at high oxidation rate of 0.4 ± 0.1 kg-N kg-VSS-1 d-1 was achieved for more than 100 days. Nitrite accumulation (N-NO2-/N-NOX) of 92% was maintained. Thus, consistently small amounts of present NOB were efficiently suppressed. Our mathematical model explained how ADCS enhanced the inhibition of NOB growth via NH3 and HNO2. This approach will produce effluent suitable for anammox even under winter conditions in mild climates.
- MeSH
- bioreaktory * MeSH
- dusík MeSH
- dusitany MeSH
- odpad tekutý - odstraňování * MeSH
- odpadní voda * MeSH
- odpadní vody MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- dusík MeSH
- dusitany MeSH
- odpadní voda * MeSH
- odpadní vody MeSH
Energy consumption of municipal wastewater treatment plants can be reduced by the anaerobic pre-treatment of the main wastewater stream. After this pre-treatment, nitrogen can potentially be removed by partial nitritation and anammox (PN/A). Currently, the application of PN/A is limited to nitrogen-rich streams (>500 mg L(-1)) and temperatures 25-35 °C. But, anaerobically pretreated municipal wastewater is characterized by much lower nitrogen concentrations (20-100 mg L(-1)) and lower temperatures (10-25 °C). We operated PN/A under similar conditions: total ammonium nitrogen concentration 50 mg L(-1) and lab temperature (22 °C). PN/A was operated for 342 days in a 4 L moving bed biofilm reactor (MBBR). At 0.4 mg O2 L(-1), nitrogen removal rate 33 g N m(-3) day(-1) and 80 % total nitrogen removal efficiency was achieved. The capacity of the reactor was limited by low AOB activity. We observed significant anammox activity (40 g N m(-3) day(-1)) even at 12 °C, improving the applicability of PN/A for municipal wastewater treatment.
- Klíčová slova
- Anammox in biofilm, Low strength wastewater, MBBR, Nitrogen removal, Partial nitritation,
- MeSH
- amoniové sloučeniny chemie MeSH
- anaerobióza MeSH
- biofilmy MeSH
- bioreaktory MeSH
- čištění vody MeSH
- dusík chemie MeSH
- hybridizace in situ fluorescenční MeSH
- kyslík chemie MeSH
- nízká teplota MeSH
- odpad tekutý - odstraňování metody MeSH
- odpadní voda chemie MeSH
- teoretické modely MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- amoniové sloučeniny MeSH
- dusík MeSH
- kyslík MeSH
- odpadní voda MeSH
