Methane (CH₄) formation in wastewater treatment is linked to long residence times under anaerobic conditions such as those in sewers and primary treatment units. Emissions of this methane to the atmosphere can occur under turbulent flows and, potentially, during aeration in an activated sludge plant. An online, 8-week monitoring campaign of CH₄emissions and operational conditions was conducted to study emissions from a full-scale nitrifying activated sludge plant (ASP). Significant emissions were found throughout the aerated lane, with the highest values observed two thirds down the lane. Emissions had high diurnal and spatial variability, with values ranging from 0.3 to 24 g CH₄/h. No significant correlations were found between dissolved oxygen, aeration or influent loads. The results suggest that emissions are linked to upstream process conditions, with potential for methane generation in-lane due periods of limited oxygen availability. The dynamic oxygen profile observed suggests that aerobic and anoxic conditions coexist in the lane, leading to limited oxygen diffusion from the bulk liquid to the inner regions of the floc where anoxic/anaerobic layers may allow methanogenic microorganisms to survive. The average emission factor was 0.07 % of removed chemical oxygen demand, giving a total of 668 kg CH₄/year and 14,000 CO₂equivalents/year. The operational carbon associated with the energy requirements of the ASP increased by 5 %. With emerging legislation requiring the reporting of greenhouse gas emissions, the carbon impact may be significant, particularly as the industry moves towards a carbon-reducing future. Therefore, an adequate profiling of full-scale emissions is critical for future proofing existing treatment technologies.

Anticoagulants are biocides widely used as pest control agents in agriculture, urban infrastructures, and domestic applications for the control of rodents. Other anticoagulants such as warfarin and acenocoumarol are also used as drugs against thrombosis. After use, anticoagulants are discharged to sewage grids and enter wastewater treatment plants (WWTPs). Our hypothesis is that WWTP effluents can be a source of anticoagulants to receiving waters and that these can affect aquatic organisms and other nontarget species. Therefore, the objective of the present study was to determine the occurrence of 11 anticoagulants in WWTPs receiving urban and agricultural wastewaters. Warfarin was the most ubiquitous compound detected in influent waters and was partially eliminated during the activated sludge treatment, and low nanograms per liter concentration were found in the effluents. Other detected compounds were coumatetralyl, ferulenol, acenocoumarol, flocoumafen, brodifacoum, bromadiolone, and difenacoum at concentrations of 0.86–87.0 ng L⁻¹. Considering water volumes of each WWTP, daily emissions were estimated to be 0.02 to 21.8 g day⁻¹, and thus, WWTPs contribute to the loads of anticoagulants to receiving waters. However, low aquatic toxicity was observed using Daphnia magna as a model aquatic organism.

Extracellular polymeric substances (EPS) are an extracellular matrix found in sludge which plays a crucial role in flocculation by interacting with the organic solids. Therefore, to enhance pretreatment of sludge, EPS have to be removed. In this study, EPS were removed with a chemical extractant, NaOH, to enhance the bacterial pretreatment. A lysozyme secreting bacterial consortium was isolated from the waste activated sludge (WAS). The result of density gradient gel electrophoresis (DGGE) analysis revealed that the isolated consortium consists of two strains. The two novel strains isolated were named as Jerish03 (NCBI accession number KC597266) and Jerish 04 (NCBI accession number KC597267) and they belong to the genus Bacillus. Pretreatment with these novel strains enhances the efficiency of the aerobic digestion of sludge. Sludge treated with the lysozyme secreting bacterial consortium produced 29 % and 28.5 % increase in suspended solids (SS) reduction and chemical oxygen demand (COD) removal compared to the raw activated sludge (without pretreatment) during aerobic digestion. It is specified that these two novel strains had a high potential to enhance WAS degradation efficiency in aerobic digestion.

The accumulation and fraction distribution of Ni(II) in sludge was determined, and their effect on the performance of sequencing batch reactor (SBR) systems was evaluated at laboratory scale. The results showed that the removal efficiencies of substrates decreased significantly with increasing feeding concentration of Ni(II) into SBRs. The concentration of Ni(II) fed into the SBRs was significantly positively correlated with the Ni(II) contents accumulated in the sludge, while it was negatively correlated with the biomass in the SBRs. The accumulated Ni(II) in the sludge was distributed mainly in the available fraction, accounting for 75.8–90.0 % of the total Ni(II) content. The accumulated content of Ni(II) in each sludge fraction could be predicted satisfactorily by the feeding Ni(II) concentrations in the solutions. As compared with the total contents and other chemical fractions, Ni(II) in the oxidizable fraction in sludge exhibited more important inhibition effects on sludge microorganisms in the SBRs.

The biological treatment of ammonia-rich landfill leachates due to an inadequate C to N ratio requires expensive supplementation of carbon from an external carbon source. In an effort to reduce treatment costs, the objective of the study was to determine the feasibility of nitrogen removal via the nitrite pathway during landfill leachate co-treatment with municipal wastewater. Initially, the laboratory-scale sequencing batch reactor (SBR) was inoculated with nitrifying activated sludge and fed only raw municipal wastewater (RWW) during a start-up period of 9 weeks. Then, in the co-treatment period, consisting of the next 17 weeks, the system was fed a mixture of RWW and an increasing quantity of landfill leachates (from 1 to 10 % by volume). The results indicate that landfill leachate addition of up to 10 % (by volume) influenced the effluent quality, except for BOD₅. During the experiment, a positive correlation (r² = 0.908) between ammonia load in the influent and nitrite in the effluent was observed, suggesting that the second step of nitrification was partially inhibited. The partial nitrification (PN) was also confirmed by fluorescence in situ hybridisation (FISH) analysis of nitrifying bacteria. Nitrogen removal via the nitrite pathway was observed when the oxygen concentration ranged from 0.5 to 1.5 mg O₂/dm³and free ammonia (FA) ranged from 2.01 to 35.86 mg N-NH₃/dm³in the aerobic phase. Increasing ammonia load in wastewater influent was also correlated with an increasing amount of total nitrogen (TN) in the effluent, which suggested insufficient amounts of assimilable organic carbon to complete denitrification. Because nitrogen removal via the nitrite pathway is beneficial for carbon-limited and highly ammonia-loaded mixtures, obtaining PN can lead to a reduction in the external carbon source needed to support denitrification.

To better construct a bioaugmented system for tobacco wastewater treatment, activated sludge was inoculated with different concentrations of the nicotine-degrading bacterium Pseudomonas sp. HF-1. The results showed that inoculum concentrations of 0.55 ± 0.01 and 1.10 ± 0.03 mg/g (dry weight of strain HF-1/dry weight of activated sludge) were best to ensure strain HF-1 survival and successful bioaugmentation. The release pattern of autoinducer (AI) for quorum sensing in the bioaugmented system was also investigated. During the period of HF-1 inoculation, compared with failed bioaugmented systems, AI-2 was significantly increased in the successful systems, suggesting that AI-2-mediated bacterial communication played an important role in the colonization of HF-1. When inoculation of strain HF-1 was stopped, the amount of AI-2 decreased and leveled out in all systems. Notably, there was a greater than threefold increase of short-chain AHLs in failed bioaugmented systems, but no increase in successful ones, implying that the fluctuation of short-chain AHLs could be an indicator of the failure of bioaugmentation. Thus, AI-2-mediated quorum sensing could be implemented to facilitate HF-1 colonization.

OECD ready biodegradability tests have been central to understanding the biodegradation of chemicals from a regulatory perspective for many decades. They are not fit for contemporary prioritisation of chemicals based on persistence, however, due to the low concentration of inocula used, short duration and high variability between tests. Two OECD standard inoculum pretreatment methods (settlement and filtration) were investigated to observe their effect on the probability of biodegradation and associated changes in bacterial community structure and diversity of inocula sourced from the activated sludge process of wastewater treatment plants. Both settlement and filtration were shown to dramatically and significantly reduce the probability and increase the variability of biodegradation of 4-nitrophenol compared to the use of unprocessed inocula. These differences were associated with a significant hundred-fold reduction in cell numbers and solids content and a significant shift in bacterial community structure that was sometimes accompanied by significant reductions in detectable operational taxonomic unit richness and evenness. The natural variation (between different environments) and variation due to differential selection of bacterial communities (by different pretreatment methods) is offered as an explanation for the historical high variability in standard OECD ready biodegradability tests.

The degradation of Romandolide® ([1-(3′,3′-dimethyl-1′-cyclohexyl)ethoxycarbonyl] methyl propanoate), a synthetic alicyclic musk, by activated sludge inocula was investigated using both the manometric respirometry test OECD 301F and the CO₂evolution test. In addition to measuring its biodegradability, key steps of the upper part of the metabolic pathway responsible for Romandolide® degradation were identified using extracts at different time points of incubation. Early metabolism of Romandolide® yielded ester hydrolysis products, including Cyclademol® (1-(3,3-dimethylcyclohexyl)ethanol). The principal metabolites after 31 days were identified as 3,3-dimethyl cyclohexanone and 3,3-dimethyl cyclohexyl acetate. Formation of 3,3-dimethyl cyclohexanone from Cyclademol® by sludge was confirmed in subsequent experiments using Cyclademol® as a substrate, indicating the involvement of an oxygen insertion reminiscent of a Baeyer–Villiger oxidation. Further mineralization of 3,3-dimethyl cyclohexanone was also confirmed in subsequent studies. Three steps were thus required for complete biodegradation of the alicyclic musk: (1) successive ester hydrolyses leading to the formation of Cyclademol® with concomitant degradation of the resulting acids, (2) conversion of Cyclademol® into 3,3-dimethyl cyclohexanone, and (3) further mineralization via ring cleavage.

Nonylphenol ethoxylates (NPEOs) with low ethoxylation degree (NPₐᵥ₂EO; containing two ethoxy units on average) and estrogenic properties are the intermediate products of nonionic surfactant NPEOs. To better understand the environmental fate of low-ethoxylated NPEOs, phylogenetically diverse low-ethoxylated NPEO-degrading bacteria were isolated from activated sludge using gellan gum as the gelling reagent. Four isolates belonging to four genera, i.e., Pseudomonas sp. NP522b in γ-Proteobacteria, Variovorax sp. NP427b and Ralstonia sp. NP47a in β-Proteobacteria, and Sphingomonas sp. NP42a in α-Proteobacteria were acquired. Ralstonia sp. NP47a or Sphingomonas sp. NP42a, have not been reported for the degradation of low-ethoxylated NPEOs previously. The biotransformation pathways of these isolates were investigated. The first three strains (NP522b, NP427b, and NP47a) exhibited high NPₐᵥ₂EO oxidation ability by oxidizing the polyethoxy (EO) chain to form low-ethoxylated nonylphenoxy carboxylates, and then further oxidizing the alkyl chain to form carboxyalkylphenol polyethoxycarboxylates. Furthermore, Sphingomonas sp. NP42a degraded NPₐᵥ₂EO through a nonoxidative pathway with nonylphenol monoethoxylate as the dominant product.

Modern society grapples with large amounts of household waste. The anaerobic digestion of this waste offers a promising source for energy-rich biogas production but generates high toxic effluents that require treatment before reuse or disposal into the environment. This study aimed to investigate three techniques, namely coagulation/flocculation, electro-coagulation, and activated sludge, in terms of efficiency in the treatment of these effluents. It also aimed to assess their toxicity effects on the germination and growth of durum wheat Triticum aestivum L. seeds before and after 6 days of treatment. Activated sludge was most efficient in reducing chemical oxygen demand, turbidity, and conductivity (95.7 %, 15.8 %, and 37.5 %, respectively). The effluent treated with this technique induced a marked delay in germination (low mean time of germination) and a significant reduction in the percentages of seed germination and root and leaf growths. It was also noted to strongly induce lipid peroxidation in roots and leaves, which presumably explained the germination/growth inhibition of the wheat seeds. The effluent also induced marked lipid peroxidation effects and strongly inhibited the activities of butyrylcholinesterase in mice bone marrows. The effluent shows a high ability to inhibit the growth of three microalgae; these endpoints are useful tools to biomonitor the physico-chemical quality of this wastewater. Overall, while no significant alterations were observed in terms of animal and vegetable toxicities when the effluent was treated by coagulation/flocculation, activated sludge treatment proved efficient in reducing the toxicities induced by the untreated effluents. The results indicate that the application of this technique is promising with regards to attaining efficient, eco-friendly, and cost-effective strategies for the management and treatment of household waste.